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    Editorial
    Validity and security of assisted reproductive technology
    ZHAO Xiao-ming
    2012, 32 (8):  965. 
    doi: 10.3969/j.issn.1674-8115.2012.08.001

    Abstract ( 1302 )   PDF (236KB) ( 1484 )  

    In vitro fertilization-embryo transfer (IVF-ET) technology is a major breakthrough for the treatment of infertility. Along with the maturation and breakthrough of derivative technologies including intracytoplasmic sperm injection and pre-implantation genetic diagnosis, IVF-ET has become one of the fastest-growing technologies in life sciences. However, the cycle pregnancy rate of IVF-ET is still hovering around 40%. How to improve the success rate of IVF is worth pondering. Meanwhile, how to reduce or avoid the occurrence of complications such as ovarian hyperstimulation syndrome and multiple gestation pregnancy is the problem that clinicians need to remain vigilant of. In IVF, the simple, economical and effective principle should be followed, and the risk prevention should be particularly concerned about in the process of improving pregnancy rate.

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    Monographic report (Infertility and assisted reproductive technology)
    Significance of analysis of karyotype of chromosome in diagnosis and treatment of male infertility and screening in sperm donors
    WANG Xiao-bo, HU Hong-liang, LIU Yong, et al
    2012, 32 (8):  968. 
    doi: 10.3969/j.issn.1674-8115.2012.08.002

    Abstract ( 1892 )   PDF (367KB) ( 1253 )  

    Objective To analyse karyotype of chromosome in patients with infertility and sperm donors, and explore its clinical significance. Methods A total of 967 infertile patients (infertility group) and 3 184 semen donors (donor group) were selected for analysis of karyotype of chromosome, and the incidence and types of abnormal karyotypes were compared between two groups. Results The incidence of abnormal karyotypes in infertility group was significantly higher than that in donor group (14.06% vs 3.39%, P<0.01). The main type of abnormal karyotype in infertility group was non-polymorphic chromosomal abnormalities, whose incidence was significantly higher than that in donor group (9.62% vs 0.25%, P<0.05). The main type of abnormal karyotype in donor group was polymorphic chromosomal abnormalities, whose incidence was not significantly different from that in infertility group (3.14% vs 4.44%, P>0.05). Besides, two abnormal karyotypes [46,XY,t(3;12)(p23;q24) and 46,XY,inv(20)(p13;q13.1)] were first reported worldwide in infertility group. Conclusion Chromosome non-polymorphism abnormalities is one of the major causes for male infertility, and karyotype analysis for infertile patients and sperm donors is of great significance to reduce birth defects.

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    Effects of vapor nitrogen freezing method and direct liquid nitrogen freezing method on motility, morphology and ultrastructure of human sperm
    LIU Feng, MENG Ying, WANG Feng, et al
    2012, 32 (8):  973. 
    doi: 10.3969/j.issn.1674-8115.2012.08.003

    Abstract ( 1168 )   PDF (720KB) ( 1200 )  

    Objective To investigate the effects of vapor nitrogen freezing method and direct liquid nitrogen freezing method on the motility, morphology and ultrastructure of post-thaw human sperm. Methods Twenty-five semen samples from semen donors were selected, and glycerol-yolk-sodium citrate (GYC) was added as cryoprotectant with the proportion of 1∶1. Each semen sample was equally distributed to two cryovials, and were cryopreserved with vapor nitrogen freezing method and direct liquid nitrogen freezing method respectively. The total motility rate, progressive motility rate and motility parameters of post-thaw sperm were examined by IVOS sperm analyzer. The morphology of post-thaw sperm was observed with papanicolaou staining, and the rate of post-thaw sperm with normal morphology was calculated. Besides, the ultrastructure of post-thaw sperm was observed by transmission electron microscope. Results It cost 468 s to drop in temperature from 28 ℃ to -185 ℃ by vapor nitrogen freezing method, and an hour later the temperature went to -196 ℃. However, it only cost 90 s to drop in temperature from 30℃ to -196 ℃ by direct liquid nitrogen freezing method. The total motility rates, progressive motility rates, motility parameters and rates of post-thaw sperm with normal morphology in two cryopreservation methods were significantly lower than those before cryopreservation (P<0.05). The revival rate, total motility rate and progressive motility rate in vapor nitrogen freezing method were significantly higher than those in direct liquid nitrogen freezing method (P<0.05). There was no significant difference in motility parameters and rates of post-thaw sperm with normal morphology between two cryopreservation methods (P>0.05). Transmission electron microscopy indicated that no obvious damage was found on the ultrastructure of serosa and acrosome membrane at the head of post-thaw sperm in two different cryopreservation methods by using GYC as cryprotectant. Conclusion GYC has a protective effect on the ultrastructure of sperm. Vapor nitrogen freezing method may yield better post-thaw results than direct liquid nitrogen freezing method in cryopreservation of human sperm.

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    Influence of temperature and Hoechst 33258 concentration on vital labeling of human sperm
    LU Hui, SHI Wen-bo, XIAO Yu-fang, et al
    2012, 32 (8):  979. 
    doi: 10.3969/j.issn.1674-8115.2012.08.004

    Abstract ( 1312 )   PDF (580KB) ( 1238 )  

    Objective To investigate the influence of culture temperature and concentration of fluorescence dye Hoechst 33258 (H258) on vital labeling of human sperm. Methods Fourteen semen samples were randomly selected, and were cultured with 0 mg/mL (control), 0.01 mg/mL and 0.1 mg/mL H258 for 1 h under different temperatures (0 ℃, room temperature and 37 ℃) after washing. Progressive motility of human sperm was assessed, and H258 staining rates of the sperm were recorded. Semen samples without treatment were served as blank controls. Results Under the conditions of this research, H258 concentrations and culture temperatures had significant impact on sperm motility. The percents of progressive motility of sperm gradually decreased with the increase of H258 concentrations and decrease of culture temperatures (P<0.01). The staining rates of the sperm gradually increased with the culture temperatures and mass concentrations of H258, and there were significant differences among staining rates of the sperm cultured with different temperatures at the mass concentration of 0.1 mg/mL (P<0.01). Conclusion H258 can be used in vital labeling of human sperm. The optimal staining condition was 37 ℃+0.1 mg/mL H258 in this study. Culture condition should be determined by specific purpose.

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    Correlation analysis between epididymal sperm DNA damage and outcome of intracytoplasmic sperm injection in patients with obstructive azoospermia
    WANG Hong-xiang, CHEN Bin, HU Kai, et al
    2012, 32 (8):  983. 
    doi: 10.3969/j.issn.1674-8115.2012.08.005

    Abstract ( 1284 )   PDF (405KB) ( 1160 )  

    Objective To investigate the epididymal sperm DNA damage in patients with obstructive azoospermia(OA), and explore its correlationship with the outcome of subsequent intracytoplasmic sperm injection (ICSI). Methods One hundred and twenty-one patients with OA suject to ICSI were selected as OA group, and 38 semen samples with normal range of parameters were served as control group. Patients in OA group underwent percutaneous epididymal sperm aspiration and microscopic analysis, while computer assisted semen analysis was conducted in control group. Acridine orange fluorescence staining tests of aspiration fluid/semen were carried out in both groups. The normal parameter of DNA fragmentation index (DFI) was below 30%, and both groups were subdivided into high DFI subgroups (DFI>30%) and low DFI subgroups (DFI<30%). The fertilization rate and clinical pregnancy rate of the subsequent ICSI in patients with OA were statistically analysed. Results The sperm density and the proportion of forward moving sperm of OA group were significantly lower than those of control group as a whole (P<0.01). There was no significant difference in the sperm density between high DFI subgroup and low DFI subgroup of OA group(P>0.05), while the proportion of forward moving sperm in high DFI subgroup of OA group was significantly lower than that in low DFI subgroup of OA group (P<0.01). The negative rate of acridine orange test in OA group was significantly lower than that in control group as a whole (P<0.01). As to the outcome of ICSI in OA group, there was no significant difference in the fertilization rate between high DFI subgroup and low DFI subgroup (P>0.05), while the clinical pregnancy rate in high DFI subgroup was significantly lower than that in low DFI subgroup (P<0.01). Conclusion It is essential to evaluate the sperm DNA for patients with OA before ICSI. Sperm DNA damage analysis for the epididymal sperm from diagnostic percutaneous epididymal sperm aspiration is an effective method for predicting ICSI outcome for patients with OA.

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    Research on ultra-rapid cryopreservation of small numbers of human spermatozoa in capillary
    LIU Yong, LIU Feng, LI Zheng, et al
    2012, 32 (8):  988. 
    doi: 10.3969/j.issn.1674-8115.2012.08.006

    Abstract ( 1274 )   PDF (318KB) ( 1006 )  

    Objective To explore sperm cryopreservation in capillary by ultra-rapid procedure, and seek an optimal method to cryopreserve small numbers of human spermatozoa. Methods Twenty semen samples from 20 healthy donors were selected from Shanghai Human Sperm Bank. Each sample was divided into 4 groups for cryopreservation after dilution to the density of (3-5)×106/mL (3-5/high power filed). In group A, samples mixed with sperm freezing medium in 0.5 mL straw were frozen by program procedure. In group B, samples mixed with sperm freezing medium in capillary were frozen by program procedure. In group C, samples mixed with modified glycerol-yolk-citrate (GYC) in capillary were frozen by program procedure. In group D, samples mixed with modified GYC in capillary were frozen by direct liquid nitrogen vapor method with ultra-rapid speed. The post-thaw viability rates and normal morphology rates were compared among groups. Results The pre-thaw viability rate of the 4 groups was (68.6±21.3)%, and the post-thaw viability rates of group A, B, C and D were (32.2±17.2)%, (41.5±18.4)%, (44.9±18.0)% and (50.2±19.7)%, respectively. The post-thaw viability rates of group B, C and D were significantly higher than that of group A (P<0.05), and the post-thaw viability rate of group D was significantly higher than those of the other 3 groups (P<0.05). There was no significant difference in the post-thaw normal morphology rates among group A, B, C and D [(14.2±4.1)%, (15.0±4.7)%, (14.5±5.2)% and (15.9±4.9)%, respectively, P>0.05]. Conclusion Ultra-rapid freezing with direct liquid nitrogen vapor in capillary is an optical method for cryopreservation of small numbers of human spermatozoa, which may yield high recovery rate. Modified GYC has the same cryoprotecting effect as sperm freezing medium.

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    Clinical significance of hysteroscopy after repeated implantation failure in in vitro fertilization and embryo transfer
    CHEN Ping-ping, GAO Min-zhi, ZHAO Xiao-ming, et al
    2012, 32 (8):  992. 
    doi: 10.3969/j.issn.1674-8115.2012.08.007

    Abstract ( 1540 )   PDF (371KB) ( 1078 )  

    Objective To investigate the clinical significance of hysteroscopy after repeated implantation failure (RIF) in in vitro fertilization and embryo transfer (IVF-ET). Methods The hysteroscopic results of 197 women with failure in IVF-ET for no less than two times (RIF group) and 176 women undergoing artificial insemination with donor semen (control group) were retrospectively analysed, and the incidences and types of abnormal hysteroscopic findings were compared between two groups. Results The incidence of abnormal hysteroscopic findings in RIF group was significantly higher than that in control group (51.78% vs 28.98%, P<0.01), and the types of abnormal hysteroscopic findings in RIF group were more than those in control group. The main types of abnormal hysteroscopic findings in RIF group were endometrial abnormalities (polypi and polypoid proliferation, 26.39%; endometritis, 4.06%) and abnormal shape of uterine cavity (uterine cavity narrowness, 9.64%; uterine cavity adhesion, 5.08%). The incidence of uterine cavity abnormalities of each age group in RIF group was higher than that in control group, especially for the age groups of 25 to 29 years and 30 to 34 years (P<0.01, P<0.05). Logistic regression analysis revealed that types of RIF (P=0.001, OR=2.320, 95%CI:1.440-3.736) and age (P=0.039, OR=1.403, 95%CI:1.016-1.935) were risk factors of uterine cavity abnormalities, while duration of infertility (P=0.747,OR=1.038,95%CI:0.827-1.303) was not risk factor of uterine cavity abnormalities. In women with RIF, there was no significant difference in the incidence of uterine cavity abnormalities between primary infertility group and secondary infertility group (P>0.05), while the incidence of polypi and polypoid proliferation in primary infertility group was significantly higher than that in secondary infertility group (31.45% vs 17.80%,P<0.05), and the incidence of uterine cavity adhesion in primary infertility group was significantly lower than that in secondary infertility group (2.42% vs 9.59%,P<0.05). Conclusion The incidence of uterine cavity abnormalities in women with RIF is significantly higher than that in normal women, and the main types of uterine cavity abnormalities are polypi and polypoid proliferation. Hysteroscopic examiation should be carried out in women with RIF to improve the environment of uterine cavity and endometrium receptivity. Women with RIF are advised to undergo hysteroscopy before receiving a second ET.

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    Preliminary investigation and analysis of demand for ethics education in patients with infertility
    GUO Jing, WANG Ling, CHEN Pei, et al
    2012, 32 (8):  997. 
    doi: 10.3969/j.issn.1674-8115.2012.08.008

    Abstract ( 1025 )   PDF (228KB) ( 994 )  

    Objective To identify the medical demand of patients with infertility and the content and implementation method of ethics education. Methods A total of 150 patients with infertility were randomly selected, and the questionnaire survey of demand for ethics education was conducted. The questionnaire was designed on the basis of expert consultation, which included the demand for medical service and difficulty in asking for medical help, current situation of ethics education, awareness of patients on treatment, content and effect of ethics education and ways of ethics education. The recovery rate of questionnaire was 100%. Results Patients with infertility needed medical advice urgently, but the current situation of ethics education provided by medical institutions was not optimistic. Patients were not completely aware of the treatment course and legal consequence, and were not clear of the content of ethics education. Besides, the degree of acceptance for the mode of ethics education also needed to be improved. Conclusion Ethics education for patients with infertility is necessary and feasible, in which strengthening informed consent principle is the key problem to perfect the current ethics education. Medical institutions should design the content and process of ethics education standardly, and implement the ethics education though different ways, so that patients with infertility can get a variety of information.

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    Timing and pregnancy outcome of artificial insemination with donor semen after failure in intracytoplasmic sperm injection
    XIA Lan, ZHAO Xiao-ming, SUN Yun, et al
    2012, 32 (8):  1000. 
    doi: 10.3969/j.issn.1674-8115.2012.08.009

    Abstract ( 1334 )   PDF (395KB) ( 941 )  

    Objective To investigate the timing and outcome of artificial insemination with donor sperm(AID) after failure in intracytoplasmic sperm injection (ICSI) in patients with severe oligoasthenospermia or obstructive azoospermia. Methods The cycle pregnancy rates of AID (93 treatment circles) after failure in ICSI in 46 infertile couples with severe oligoasthenospermia or obstructive azoospermia were retrospectively analysed, and were compared with those directly undergoing AID during the same period (AID group, n=74). Besides, 1 007 patients undergoing ICSI during the same period (1 128 treatment cycles) were selected, and were divided into ejaculated sperm group and epididymal sperm group according to different sources of sperm, and were also divided into mild oligoasthenoteratozoospermia group (group A, n=157), severe oligoasthenoteratozoospermia group (group B, n=305), testicular or epididymal aspiration group (group C, n=110) and the other aspiration group (group D, n=435) according to sperm parameters. The fertility rate, cleavage rate, excellent embryo rate and pregnancy rate were compared among group A, B, C and D, and the fertility rate, cleavage rate, excellent embryo rate and pregnancy rate of the first to fourth cycle of ICSI in group B were compared. Results The cumulative pregnancy rate of 46 patients who failed in multi-cycle ICSI treatment and transferred to 1 to 4 cycles of AID reached 52.2%, and the cycle pregnancy rate was 25.8%, which was not significantly different from the cycle pregnancy rate of AID control group (27.6%)(P>0.05). The fertility rate (75.3%) and pregnancy rate (28.7%) of group C were significantly lower than those of group A, B and D (P<0.01), the excellent embryo rate of group B (40.7%) were lower than group A, C and D, and the pregnancy rate of group B (35.1%) were significantly lower than those of group A (P<0.01) and group D (P<0.05). The pregnancy rates of the third cycle and fourth or above cycle of ICSI were 15.4% and 0 in group B, which were significantly lower than those of the first cycle and second cycle (P<0.05). Conclusion AID can be performed after informed consent in patients with failure in more than 3 cycles of ICSI or with severe asthenozoospermia and obstructive azoospermia whose sperm is few, and a high pregnancy rate can be achieved.

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    Study on reactive oxygen species and total antioxidant capacity of serum and follicluar fluid in in vitro fertilization cycles
    NIU Zhi-hong, WU Dan-dan, LIN Nan, et al
    2012, 32 (8):  1005. 
    doi: 10.3969/j.issn.1674-8115.2012.08.010

    Abstract ( 1170 )   PDF (348KB) ( 1224 )  

    Objective To investigate the characteristics of reactive oxygen species (ROS) and total antioxidant capacity (TAC) of serum and follicular fluid in in vitro fertilization (IVF) cycles. Methods Patients (245 cycles) undergoing IVF were enrolled. Blood samples of early follicle phase and ovum retrieval day and follicular fluid of ovulation phase were collected, ROS and TAC were examined, and their relationship with clinical characteristics was analysed. Results Serum ROS on ovum retrieval day was significantly higher than that in early follicle phase [(77.5±62.3) cps vs (59.8±54.6) cps, P<0.05], while there was no significant difference in TAC (P>0.05). There was positive correlation between follicular ROS and serum ROS on ovum retrieval day, and between follicular TAC and serum TAC of early follicle phase and ovum retrieval day (P<0.05). There was no correlation of follicular ROS and TAC with age, peak estradiol (E2), oocyte number, peak E2/oocyte number and fertilization rate. However, follicular ROS was positively related to body mass index (BMI)(r=0.29,P=0.04). Follicular ROS in patients with BMI<19 kg/m2 was significantly lower than those in patients with BMI 23-25 kg/m2 and those with BMI>25 kg/m2 (P<0.05). Conclusion Serum ROS in ovulation phase is higher than that in early follicular phase, follicular ROS has a positive correlation with BMI.

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    Research progress of effects of PDE5 inhibitors on sperm motility and fertilizing capacity
    LU Ke-min, LU Hui, LI Zheng
    2012, 32 (8):  1010. 
    doi: 10.3969/j.issn.1674-8115.2012.08.011

    Abstract ( 1668 )   PDF (331KB) ( 1252 )  

    Phosphodiesterase-5 (PDE5) inhibitors are commonly used in the treatment of penile erectile dysfunction (ED). Recent studies have revealed that PDE5 inhibitors have impact on sperm motility and fertilizing capacity, and can significantly improve sperm motility in the appropriate concentration. The effects of PDE5 inhibitors on sperm motility and fertilizing capacity are reviewed in this paper.

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    Research progress of endometrial receptivity markers
    ZHANG Ping-gui, FENG Yun
    2012, 32 (8):  1014. 
    doi: 10.3969/j.issn.1674-8115.2012.08.012

    Abstract ( 1548 )   PDF (474KB) ( 1378 )  

    The low implantation rate is one of the major issues in the field of reproductive medicine, which has not been solved nowadays, and it also remains the major rate-limiting step in the development of human-assisted reproductive technology. Successful implantation requires a vital embryo and receptive endometrium. However, a large number of clinical trials have found that even if the embryo quality is good, the embryo implantation rate is continued low. The endometrium receptivity damage may be the main factor leading to low implantation rate. The better establishment of endometrial receptivity state may promote embryo-endometrial cross-talk, allowing the embryo implantation. The identification of endometrial receptivity markers is a prerequisite for improving endometrial receptivity. In recent years, the assessment of endometrial receptivity has become a research hotspot in reproductive medicine. The endometrial receptivity markers such as pinopodes, leukemia inhibitory factor (LIF) and homeobox A10 (HOXA10) gene are introduced in this paper.

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    A new technique of mouse intracytoplasmic sperm injection without assistance of Piezo for high survival rate of oocytes
    MIAO Cong-xiu, YAN Zhi-guang, YANG Hong-mei, et al
    2012, 32 (8):  1020. 
    doi: 10.3969/j.issn.1674-8115.2012.08.013

    Abstract ( 1464 )   PDF (520KB) ( 1083 )  

    Objective To achieve higher survival rate of mouse oocytes through modification of established technique of mouse intracytoplasmic sperm injection (ICSI) without assistance of Piezo. Methods The step of deeply sucking and deeply insertion to mouse oocytes in the established method (one-step method without Piezo-ICSI group) was modified to two steps (two-step method without Piezo-ICSI group), in which slightly sucking and slightly insertion was first performed before deeply sucking and deeply insertion. Besides, oocytes of parthenogenetic activation were served as control group. The survival rate, fertilization rate and cleavage rate of oocytes were compared between one-step method without Piezo-ICSI group and two-step method without Piezo-ICSI group. Results The survival rate of oocytes in two-step method without Piezo-ICSI group was significantly higher than that in one-step method without Piezo-ICSI group (92.2% vs 83.7%, P<0.01), while there was no significant difference in both fertilization rate and cleavage rate of oocytes between two group (P>0.05). Conclusion Some oolemma becomes fragile when the shape of the oocyte membrane is changed by drawing deeply with new type of holding pipette. The new technique of mouse ICSI avoids this sensitive fragile by two-step method for oocyte holding and insertion, which is beneficial to achieve higher survival rate of mouse oocytes.

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    Original article (Basic research)
    Influence of blocking Notch signaling pathway by γ-secretase inhibitor on proliferation of lung adenocarcinoma A549 cells
    ZHOU Min, FAN Zhi-wen, HAN Rui-chao
    2012, 32 (8):  1024. 
    doi: 10.3969/j.issn.1674-8115.2012.08.014

    Abstract ( 1193 )   PDF (427KB) ( 1373 )  

    Objective To observe the influence of blocking Notch signaling pathway by γ-secretase inhibitor MW167 on the proliferation of lung adenocarcinoma A549 cells. Methods Lung adenocarcinoma A549 cells were cultured in vitro under normoxic condition, and the expression of Notch receptor subtypes was detected by Western blotting and RT-PCR. Under normoxic condition, lung adenocarcinoma A549 cells were treated with different concentrations (5, 10 and 20 μmol/L) of MW167 (treatment groups). Forty-eight hours after treatment, the expression of Notch1 protein was determined by Western blotting, the expression of Notch signaling pathway downstream gene HES1mRNA was detected by RT-PCR, and the influence of MW167 on cell proliferation was determined by MTT. Besides, DMSO control groups of corresponding concentrations were established. Results Notch1-4 subtypes were expressed in A549 cells. Compared with control groups, the expression of Notch1 protein was higher (P<0.05) and that of HES1 mRNA was lower (P<0.05) in treatment groups 48 h after treatment, and the inhibition rate of HES1 mRNA expression increased with MW167 concentrations (r=0.927, P<0.01). The absorbance of each treatment group was higher than that of each control group (P<0.05), and the cell proliferation rates increased with MW167 concentrations (r=0.904, P<0.01). Conclusion Under normoxic condition, MW167 may block Notch signaling pathway and promote proliferation of A549 cells. Notch1 signaling pathway may play a tumor suppressing role in A549 cells.

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    Experimental study of esophagus-jejunum stent bypass in treatment of type 2 diabetes mellitus
    FU Suo-lin, ZHU Hui-ming, ZHENG Li, et al
    2012, 32 (8):  1029. 
    doi: 10.3969/j.issn.1674-8115.2012.08.015

    Abstract ( 1090 )   PDF (454KB) ( 1199 )  

    Objective To observe the effect of endoscopic esophagus-jejunum stent bypass (combined with laparotomy) on type 2 diabetes mellitus (T2DM) complicated with obesity in model dogs. Methods Five model dogs with T2DM complicated with obesity (experiment group) underwent endoscopic esophagus-jejunum stent bypass (combined with laparotomy). Before treatment and 4 weeks, 8 weeks and 12 weeks after treatment, fasting plasma glucose and fasting insulin were measured, intravenous glucose tolerance test (IVGTT) was performed, and β cell function and insulin resistance were evaluated with homeostasis model assessment (HOMA-β and HOMA-IR). Besides, another 5 normal Beagle dogs undergoing endoscopy and exploratory laparotomy were served as control group. Results Four weeks after treatment, the fasting plasma glucose, IVGTT-2 h plasma glucose, fasting insulin and IVGTT-2 h insulin were significantly lower than those before treatment in experiment group (P<0.01). There was no significant difference in fasting plasma glucose between experiment group and control group 12 weeks after treatment (P>0.05). HOMAIR gradually decreased (P<0.01) and HOMA-β gradually increased (P<0.01, P<0.05) in experiment group after treatment. There was no significant difference in HOMA-IR and HOMA-β between experiment group and control group 12 weeks after treatment (P>0.05). Conclusion Endoscopic esophagus-jejunum stent bypass (combined with laparotomy) can simulate and improve classic Roux-en-Y gastric bypass in model dogs with T2DM complicated with obesity, and can treat T2DM effectively and safely.

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    Promoter methylation of human RUNT-related transcription factor 3 gene in pancreatic carcinoma and its clinical significance
    LIN Zhi-chuan, CHEN Tan-gen, LI Jian-guo
    2012, 32 (8):  1034. 
    doi: 10.3969/j.issn.1674-8115.2012.08.016

    Abstract ( 1256 )   PDF (387KB) ( 1118 )  

    Objective To detect the promoter methylation of human RUNT-related transcription factor 3 (RUNX3) gene in pancreatic carcinoma, and explore its clinical significance. Methods The methylation of promoter CpG island of 14 samples of normal pancreatic tissues, 56 samples of pancreatic carcinoma and adjacent tissues, 3 strains of pancreatic carcinoma cell lines (PANC1, CFPAC-1 and SW1990) and one strain of normal liver cell line (HL-7702) was detected by methylation-specific polymerase chain reaction (MSP). The expression of RUNX3 mRNA in pancreatic carcinoma cell lines before and after treatment with methylation inhibitor 5-Aza-2 deoxycytidine (5-Aza-cdR) was detected. The relationship between abnormal promoter methylation of RUNX3 gene and clinical characteristics of pancreatic carcinoma was analysed. Results Abnormal CpG island methylation in RUNX3 gene promoter was found in 29 cases of pancreatic carcinoma (29/56, 51.79%) and 6 cases of adjacent tissues (6/56, 10.71%), while no abnormal CpG island methylation of RUNX3 gene was found in normal pancreatic tissues. Abnormal CpG island methylation of RUNX3 gene was significantly related to differentiation grade (r=0.314, P=0.018) and lymph node metastasis (r=0.370, P=0.005). There was no expression or low expression of RUNX3 mRNA in pancreatic carcinoma cell lines (PANC1, CFPAC-1 and SW1990) before treatment with 5-Aza-cdR, while there was expression of RUNX3 mRNA in pancreatic carcinoma cell lines after treatment with 5-Aza-cdR. Conclusion Abnormal CpG island methylation of RUNX3 gene is found in pancreatic carcinoma tissues and pancreatic carcinoma cell lines. Promoter hypermethylation of RUNX3 is an important mechanism for low expression of RUNX3 in pancreatic carcinoma, and is significantly related to the differentiation grade and node metastasis.

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    Effect of melatonin on acrylamide-induced DNA damage in rat testicular cells
    WU Hong-juan, CHEN Yu, ZHAO Qi, et al
    2012, 32 (8):  1038. 
    doi: 10.3969/j.issn.1674-8115.2012.08.017

    Abstract ( 1270 )   PDF (780KB) ( 1195 )  

    Objective To investigate the protective effects of melatonin (MT) on acrylamide (AA)-induced DNA damage in rat testicular cells. Methods Forty male SD rats aged 8 weeks were randomly divided into blank control group, MT group, AA group and MT+AA group, with 10 rats in each group. MT was intraperitoneally injected at the dose of 10 mg·kg-1·d-1, AA was orally administrated at the dose of 25 mg·kg-1·d-1, and the treatment lasted for 30 d. Comet assay and Comet Assay Software Project were  employed to examine the DNA damage (comet tail length, tail DNA%, tail moment and Olive tail moment) of testicular cells, TUNEL was used to detect the cell apoptosis, the levels of 8-hydrxydeoxyguanosine (8-OHdG) in testis tissues were measured by ELISA, the levels of malondialdehyde (MDA) were measured by thiobarbituric acid method, and the activity of superoxide dismutase (SOD) was determined by xanthine oxidase method. Results Compared with blank control group, the comet tail length, tail DNA%, tail moment and Olive tail moment in testicular cells significantly increased, the number of apoptotic cells significantly increased, the levels of 8-OHdG and MDA in testis tissues significantly increased, and the activity of SOD significantly decreased in AA group (P<0.05). Compared with AA group, the DNA damage of testicular cells significantly decreased, the number of apoptotic cells significantly decreased, the levels of 8-OHdG and MDA in testis tissues significantly decreased, and the activity of SOD significantly increased in MT+AA group (P<0.05). Conclusion AA could induce DNA damage and apoptosis in rat testicular cells, and cause oxidative damage in testis. However, MT could prevent the alterations caused by AA. Oxidative damage is probably one of the mechanisms of AA-induced DNA damage in rat testicular cells.

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    Olfactory dysfunction of human α-synucleinA53T transgenic mice in simulation of early symptoms of Parkinson´s disease
    ZHANG Su-fang, LI Li-xi, NI Jun, et al
    2012, 32 (8):  1043. 
    doi: 10.3969/j.issn.1674-8115.2012.08.018

    Abstract ( 1409 )   PDF (786KB) ( 1199 )  

    Objective To examine the olfactory function of human α-synucleinA53T transgenic mice, and establish a model for olfactory dysfunction of early Parkinson´s disease. Methods Human α-synucleinA53T transgenic (TG) mice of different ages and their wildtype (WT) littermates were selected. Rotarod test was used to examine the voluntary motion of TG mice aged 10 months, and DAB method was employed to observe the dopaminergic neurons in substantia nigra in mice aged 10 months for identification of motor function. Odor discrimination and habituation tests were used to observe the short-term memory and habituation of familiar scents and identification of novel scents in mice. Long-term memory test with varied intervals was employed to examine the memory of exposed scents. Besides, buried pellet test was used to investigate the perception on scents of food, which reflected the odor threshold. Results Rotarod test and observation of dopaminergic neurons indicated that the voluntary motion in TG mice aged 10 months did not change. TG mice aged 6 months exhibited subtle deficit in odor discrimination, and there was no significant difference between the time of discrimination of novel scents and that of familiar scents (P=0.120). TG mice aged 10 months exhibited more significant deficit in discrimination of scents (P=0.295). The time for finding food in TG mice aged 6 months was longer than that in WT mice (P=0.015). The short memory and habituation of mice of different ages were normal, while TG mice aged 9 months exhibited decrease in long-term memory (60 min, 80 min and 100 min of test intervals). Conclusion Human α-synucleinA53T transgenic mice exhibit deficiency in olfaction before motion function alterations, including the aspects of discrimination, memory and perception of scents, which can well simulate the early olfactory disfunction in Parkinson´s disease.

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    Effects of low protein diet supplement with α-keto acid |on oxidative stress in rats with chronic renal failure
    LONG Quan, ZHU Chun, SHAN Jian-ping, et al
    2012, 32 (8):  1050. 
    doi: 10.3969/j.issn.1674-8115.2012.08.019

    Abstract ( 1033 )   PDF (458KB) ( 1239 )  

    Objective To investigate the effects of low protein diet supplement with α-keto acid on oxidative stress in rats with chronic renal failure. Methods Forty male SD rats were randomly divided into control group (nephrocapsectomy, n=10) and model group (two-step subtotal nephrectomy, n=30). Rats in model group were subdivided into normal protein diet group (NPD group, n=10, 18% casein protein), low protein diet group (LPD group, n=10, 6% casein protein) and α-keto acid supplemented low protein group (LK group, n=10, 5% casein protein+1% α-keto acid) according to different ways of feeding one week later. Rats in control group were fed with 18% casein protein. The blood and urine parameters including serum creatinine (Scr), blood urea nitrogen (BUN), albumin (Alb), total protein (TP) and 24 h urinary protein excretion were detected 12 weeks later. Spectrophotography was used to measure levels of malonaldehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in serum and homogenate. The activity of renin and angiotensinⅡ (AngⅡ) in homogenate was determined by radioimmunoassay, and the activity of AngⅡ in plasma was detected by ELISA. Results There was no significant difference in the body weight, Alb and TP between each model subgroup and control group after 12 weeks of feeding (P>0.05). After 4, 8 and 12 weeks of feeding, 24 h proteinuria excretion in each model subgroup were significantly higher than those in control group (P<0.05), 24 h proteinuria excretion in LPD group and LK group were significantly lower than those in NPD group (P<0.05), and 24 h proteinuria excretion in LK group were significantly lower than those in LPD group (P<0.05). The activity of renin and AngⅡ in homogenate in LK group was significantly lower than that in NPD group (P<0.05). Compared with control group, the levels of MDA both in serum and homogenate in model subgroups were significantly higher (P<0.05), the levels of SOD and GSH-Px both in serum and homogenate in model subgroups were significantly lower (P<0.05), the levels of MDA in LPD group and LK group were significantly lower than that in NPD group(P<0.05), and the levels of SOD and GSH-Px in LPD group and LK group were significantly higher than those in NPD group (P<0.05). Correlation analysis revealed that the level of MDA in kidney homogenate was positively related to the activity of AngⅡ (r=0.892, P<0.001), and the levels of SOD and GSH-Px in kidney homogenate were negatively related to the activity of AngⅡ (r=-0.978, P<0.001; r=-0.965, P<0.001). Conclusion Renoprotective effects of reduced oxidative stress and proteinuria are exhibited by low protein diet supplement with α-keto acid therapy in rats with chronic renal failure without malnutrition. The favorable effects may be directly related to the nutrition therapy itself, or may be indirectly related to the suppression of intrarenal renin-angiotensin system activity.

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    Role of SUMO-specific protease 1 in pathogenesis of atherosclerosis
    HUANG Xian, WU Ji, CHENG Jin-ke
    2012, 32 (8):  1056. 
    doi: 10.3969/j.issn.1674-8115.2012.08.020

    Abstract ( 1319 )   PDF (557KB) ( 1263 )  

    Objective To investigate the potential role of SUMO-specific protease 1 (SENP1) in the pathogenesis of atherosclerosis in mice. Methods The development of atherosclerosis in SENP1+/+ X Apoe-/- mice (n=5) and SENP1+/- X Apoe-/- mice (n=4) fed with high cholesterol and high fat diet was observed with whole aorta and aorta root oil red O staining and aorta root immunohistochemical staining of macrophage marker MOMA-2. Acetylated low density lipoprotein (ac-LDL) was used to induce the formation of foam cells and oil red O staining, and the capacities of foam cell formation were compared between RAW264.7 si-ns and RAW264.7 si-SENP1. The relative expression of fatty acid-binding protein 4 (FABP4) mRNA and protein in RAW264.7 si-ns and RAW264.7 si-SENP1 was detected by Real-Time PCR and Western blotting. Results Oil red O staining indicated that after feeding with high cholesterol and high fat diet, the relative atherosclerotic lesion areas (lesion areas/total vessel areas) and aorta arch lesion areas of SENP1+/- X Apoe-/- mice were significantly larger than those of SENP1+/+ X Apoe-/- mice [(6.716±1.442) % vs (5.952±2.332)% and (364 249±45 838) μm2 vs (273 486±158 814) μm2]. Immunohistochemical staining revealed that the macrophage area/plaque area in aorta root in SENP1+/- X Apoe-/- mice and SENP1+/+ X Apoe-/- mice were (41.00±0.05)% and (40.47±0.07)% respectively, and there was no significant difference between them (P=0.954). Real-Time PCR, Western blotting and oil red O staining demonstrated that FABP4 level and capacity of foam cell formation of RAW264.7 si-SENP1 were significantly higher than those of RAW264.7 si-ns. Conclusion SENP1 may suppress the expression of FABP4 and capacity of foam cell formation of macrophages, thus may inhibit the development of atherosclerosis.

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    Original article (Clinical research)
    Incidence of abdominal aortic calcification and its relationship with cardiovascular diseases in patients undergoing maintenance hemodialysis
    CAI Hong, YAN Yu-cheng, LU Ren-hua, et al
    2012, 32 (8):  1062. 
    doi: 10.3969/j.issn.1674-8115.2012.08.021

    Abstract ( 1416 )   PDF (415KB) ( 1428 )  

    Objective To determine the incidence of abdominal aortic calcification (ACC) and its relationship with cardiovasculare diseases (CVD) in patients undergoing maintenance hemodialysis (MHD). Methods One hundred and sixty-four patients with chronic kidney disease of stage 5D undergoing MHD were selected as study objective. The parameters of renal function, fasting blood glucose, blood lipid, serum high-sensitivity C-reactive protein (hs-CRP), blood calcium, blood phosphorus, total iron binding capacity, serum intact parathyroid hormone (iPTH) and serum ferritin (SF) were measured, the urea clearance index Kt/V was calculated, and abdominal aorta calcification (AAC) score was assessed with abdominal lateral X-ray examination. The incidence, distribution characteristics and severity of AAC were analysed, the relationship between AAC and clinical parameters was explored, the incidences of CVD were compared between patients with calcification and those without calcification, and area under the receiver operating characteristic curve (AUCROC) was used for evaluation of accuracy of AAC in diagnosis of CVD in patients undergoing MHD. Results The incidence of AAC was 68.9%. The incidence of calcification and degree of calcification increased from L1 to L4 (P<0.05, P<0.001). Correlation analysis indicated that AAC was positively related to age(r=0.290,P<0.001), dialysis vintage (r=0.225,P=0.004), iPTH (r=0.342,P=0.043) and calcitrol dosage (r=0.21, P=0.006). The incidence of CVD in patients with calcification was significantly higher than that in patients without calcification (52.2% vs 29.4%,P=0.007). AAC score in patients with CVD was significantly higher than that in patients without CVD (P<0.001). AUCROC of AAC in diagnosis of CVD was 0.658 (95%CI 0.573-0.742, P<0.001). Conclusion The incidence of ACC is higher in patients undergoing MHD. ACC develops from the distal part to the proximal part, and the severity of ACC increases from the proximal part to the distal part. The incidence of ACC is related to the development of CVD.

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    Application of 18F-FDG PET/CT in patients with negative findings in serum Tg measurement and 131I-whole body scan after ablation of remaining thyroid tissues for differentiated thyroid carcinoma
    CHENG Wei-wei, WANG Hui, FENG Fang, et al
    2012, 32 (8):  1068. 
    doi: 10.3969/j.issn.1674-8115.2012.08.022

    Abstract ( 1056 )   PDF (558KB) ( 1104 )  

    Objective To investigate the application of 18F-FDG PET/CT in detection of tumor  recurrence and metastasis among patients with negative findings in serum thyroglobulin (Tg) measurement and 131I-whole body scan (131I-WBS) after total thyroidectomy and 131I ablation of the remaining thyroid tissues for differentiated thyroid carcinoma (DTC). Methods Forty-one patients with DTC underwent total thyroidectomy and 131I ablation of the remaining thyroid tissues, and were followed up with serum Tg measurement, 131I-WBS, neck ultrasound, chest CT and whole body 18F-FDG PET/CT. Results All these 41 patients had negative findings in serum Tg measurement and 131I-WBS, while abnormal lesions were detected by neck ultrasound or chest CT. Twenty-three patients had positive findings in 18F-FDG PET/CT, 3 of whom were confirmed as thyroid carcinoma metastases by pathological examinations. There was no significant difference between the size of 18F-FDG PET/CT positive lesions and that of 18F-FDG PET/CT negative lesions. Conclusion 18F-FDG PET/CT may demonstrate suspicious positive lesions in patients with negative findings in serum Tg measurement and 131I-WBS after total thyroidectomy and 131I ablation of the remaining thyroid tissues for DTC, which may help in the detection of tumor recurrence and metastasis.

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    Application of dezocine combined with propofol in anesthesia for painless gastroscopy in elderly patients
    CHENG Zhi-jun, YOU Xin-min, JI Hui, et al
    2012, 32 (8):  1072. 
    doi: 10.3969/j.issn.1674-8115.2012.08.023

    Abstract ( 1143 )   PDF (229KB) ( 1150 )  

    Objective To evaluate the effectiveness of dezocine combined with propofol in anesthesia for painless gastroscopy in elderly patients. Methods One hundred and forty-seven elderly patients undergoing painless gastroscopy were randomly assigned to dezocine group (D group, n=51, infusion with dezocine 20 μg/kg + infusion with propoful 1 mg/kg 5 min later), sufentanil group (S group, n=46, infusion with sufentanil 0.1 μg/kg+ infusion with propoful 1 mg/kg 5 min later) and control group (C group, n=50, infusion with normal saline+ infusion with propoful 1 mg/kg 5 min later). The mean arterial pressure (MAP), heart rate (HR), respiratory rate (RR) and pulse oxygen saturation (SpO2) were obtained before infusion (T1), after disappearance of eyelash reflex (T2) and after recovery of orientation (T3), and the adverse events, time for regaining consciousness and propofol dosage were also recorded. Results MAP at the time point of T2 was significantly lower than that at the time point of T1 in each group (P<0.05), and the decrease was alleviated in D group compared with C group and S group (P<0.05). HR was significantly decreased in C group and D group at the time point of T2, and the decrease in D group was more significant than that in C group (P<0.05). RR and SpO2 were significantly decreased in C group at the time point of T2, and SpO2 in C group was significantly lower than that in S group and D group at the time point of T2 (P<0.05). Time for regaining consciousness and propofol dosage in S group and D group were significantly lower than those in C group (P<0.05), and there were significant differences between D group and S group (P<0.05). Conclusion The application of dezocine combined with propofol in anesthesia for painless gastroscopy in elderly patients may significantly reduce the dosage of propofol, stabilize the respiratory and circulatory function, and have favorable safety.

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    Minimally-invasive surgical treatment of 58 patients with epidural hematoma straddling transverse sinus
    LU Zhao-feng, QIU Yong-ming, CHENG Xiao-bing, et al
    2012, 32 (8):  1075. 
    doi: 10.3969/j.issn.1674-8115.2012.08.024

    Abstract ( 1037 )   PDF (704KB) ( 1154 )  

    Objective To investigate the therapeutic effect of minimally-invasive drainage on epidural hematoma straddling transverse sinus. Methods Fifty-eight patients with traumatic epidural hematoma straddling transverse sinus and preoperative Glasgow Coma Score (GCS) 8 to 12 points were managed with minimally-invasive drainage under CT location, and 50 000 to 60 000 IU of urokinase was used for irrigation and drainage. CT scanning was performed 1 h, 3 d and 7 d after operation, volumes of hematoma were calculated, and Glasgow Outcome Score (GOS) was evaluated 3 months after operation. Results The volumes of hematoma 1 h and 3 d after operation were (20±5) mL and (15±2) mL respectively, and were significantly lower than that before operation [(45±15) mL](P<0.05). The hematoma was totally removed 3 to 5 d after operation. GCS was (12±1) points 5 d after operation, which was significantly higher than that before operation [(8±4) points](P<0.05). Three months after operation, good recovery (GOS 5 points) was achieved in 45 patients (78%), and moderate disability (GOS 4 points) was found in 9 patients (16%). Conclusion Minimally-invasive drainage is effective in the treatment of traumatic epidural hematoma straddling transverse sinus with GCS≥8 points.

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    Original article (Public health administration)
    Development and analysis of reliability and validity of Nurse Innovative Behavior Scale
    BAO Ling, WANG Lin, ZHANG Ya-qing
    2012, 32 (8):  1079. 
    doi: 10.3969/j.issn.1674-8115.2012.08.025

    Abstract ( 1419 )   PDF (373KB) ( 1316 )  

    Objective To develop a scale for measurement of nurse innovative behavior. Methods Nurse innovative behavior was defined on the basis of literature review. Three dimensions and 15 items were formulated by semi-structured interview among 12 nurses, and the items were screened with expert evaluation. Two hundred and thirty-four nurses in 2 hospitals in Shanghai were investigated with the scale, item analysis and exploratory factor analysis were conducted, and the reliability and validity of the scale were examined. Results Two hundred and six effective questionnaires were recovered, with the recovery rate of 88.03%. Through expert evaluation, item analysis and exploratory factor analysis, a 10-item Nurse Innovative Behavior Scale was developed, which included 3 factors (idea generation, support obtaining and idea realization) and was consistent with theoretical construct. These three factors accounted for 72.012% of the accumulated variances. The correlation coefficients between dimensions and total scale were higher than those between dimensions. The content validity index was 0.91, and the Cronbach´s α was 0.879. Conclusion Nurse Innovative Behavior Scale has a good reliability and validity, providing a tool to measure nurse innovative behavior.

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    Research on internal driving force of doctors in writing SCI papers
    WANG Qing, LIU Jia-zhen, FEI Xin, et al
    2012, 32 (8):  1083. 
    doi: 10.3969/j.issn.1674-8115.2012.08.026

    Abstract ( 1196 )   PDF (326KB) ( 1138 )  

    Objective To investigate the influencing factors of publishing Science Citation Index (SCI) papers in doctors, and explore the initial power in affecting SCI paper publication. Methods Three hundred and thirty-five doctors were randomly selected from 857 doctors in 40 departments of Renji Hospital affiliated to Shanghai Jiaotong University School of Medicine for questionnaire survey, and 309 effective questionnaires were recovered, with the effective rate of 92.24%. The contents of questionnaire included the situation of writing SCI papers between 2007 and 2011, the awareness and reaction to SCI policies, the internal driving force of writing SCI papers and the attitude towards scientific research required by post. Results The main reasons for writing SCI papers were professional title assessment (69.11%), department assessment (47.15%) and talent scheme application (47.15%), and the main reasons against writing SCI papers were heavy clinical workload (82.80%) and low acceptance rate of manuscript (59.14%). More than 65% doctors knew the policies about writing SCI papers, and the main source of information was dean of the department. More doctors thought that SCI papers should not serve as the standard in professional title assessment (P<0.05), while more doctors considered that SCI papers may be included in department assessment, bonus release, academic degree application, tutor qualification application and talent scheme application (P<0.05).  Scientific research required by post was found in 66.67% of respondents, and that was found in colleagues of 80.26% of respondents. Conclusion The internal driving force of doctors in writing SCI papers largely comes from the policy guidance, but most doctors are not favor of taking SCI papers as the standard of professional title assessment. Scientific research required by post is common, and the initial interest of doctors should be stimulated to improve the quality and quatity of SCI papers.

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    Review
    Research progress of translational medicine based on tumor molecular biomarkers
    LI Jian-fang, ZHU Zheng-gang, LIU Bing-ya
    2012, 32 (8):  1088. 
    doi: 10.3969/j.issn.1674-8115.2012.08.027

    Abstract ( 1340 )   PDF (332KB) ( 1433 )  

    Recently, the concept of translational medicine has come into being with the fast development of basic medicine such as molecular biology and change of medical mode. The aim of translational medicine is to quickly and effectively translate the basic research achievement of medical biology into theories, technologies, methods and drugs for clinical application, which paves a way from bench to bedside. With the fast research progress of tumor molecular biomarkers, more and more findings have been used for tumor diagnosis, treatment, prognosis and curative effect surveillance, which is an important embodiment of translational medicine.

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    Research progress of intravenous immunoglobulin application in renal transplantation
    ZHAN Jia-ming, WANG Xiang-hui
    2012, 32 (8):  1092. 
    doi: 10.3969/j.issn.1674-8115.2012.08.028

    Abstract ( 1471 )   PDF (389KB) ( 1136 )  

    The intravenous immunoglobulin (IVIG) application in renal transplantation is divided into two aspects, immunoregulation of inflammation and immune replacement therapy. The former is mainly used for the desensitization therapy, treatment of antibody-mediated rejection and treatment of steroid-resistant rejection, and the latter is mainly used for the prevention and treatment of hypogammaglobulinemia and infectious complications after renal transplantation. IVIG not only can prevent infection through sterile immunity, but can effectively regulate immune function as well. IVIG is bound to play an increasingly important role in the improvement of prognosis of clinical renal transplantation as its unique feature.

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    Neuregulin1/erbB and neuropathic pain
    ZENG Lu-lu, WANG Jing, ZHANG Xin, et al
    2012, 32 (8):  1097. 
    doi: 10.3969/j.issn.1674-8115.2012.08.029

    Abstract ( 1144 )   PDF (309KB) ( 1052 )  

    Neuropathic pain is a common chronic pain, and has received more attention recently due to its complicated pathogenesis and ineffective treatment. Many studies have found that a number of cytokines and multiple signaling pathways are involved in the pain procession after never injury. The role of Neuregulin1/erB signaling pathway in neuropathic pain is discussed in this paper.

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    Research and application of target therapy for osteoporosis
    WANG Ying, YANG Yun-mei
    2012, 32 (8):  1101. 
    doi: 10.3969/j.issn.1674-8115.2012.08.030

    Abstract ( 1358 )   PDF (380KB) ( 1175 )  

    Osteoporosis is a common disease which is caused by a systemic impairment of bone mass and microarchitecture. The disease is likely to occur in the elderly, especially in postmenopausal women. Along with the understanding of the detailed molecular biology of bone cells, the signaling network of the communication between bone-forming osteoblasts and bone-resorbing osteoclasts has been revealed, which has led to the identification of novel therapeutic targets. Novel treatment strategy aims to inhibit excessive bone resorption and increase bone formation. The most promising novel drugs include denosumab (a monoclonal antibody for a key osteoclast cytokine), odanacatib (a specific inhibitor of the osteoclast protease cathepsin K) and two endogenous inhibitors of bone formation (antibodies against the proteins sclerostin and dickkopf-1). This review will discuss the underlying physiology of novel therapies and explore their further applications.

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    Technique and method
    Isolation, culture and identification of rat articular cartilage stem cells
    TONG Wen-xue, XIANG Sheng-nan, ZHANG Ning, et al
    2012, 32 (8):  1106. 
    doi: 10.3969/j.issn.1674-8115.2012.08.031

    Abstract ( 1124 )   PDF (740KB) ( 1107 )  

    Objective To isolate and culture rat articular cartilage stem cells (ACSCs), and identify their characteristics. Methods ACSCs were isolated from rat articular cartilages by fibronectin-conglutination assay. The expression of positive stem cell surface markers (CD90 and CD44) and negative stem cell surface markers (CD45, CD31 and CD34) in ACSCs was detected by flow cytometry. The single cell colony-forming efficiency of ACSCs was determined by single cell colony-forming assay. The multidirectional differential potential of ACSCs was identified by osteogenesis, adipogenesis and chondrogensis experiment. Results ACSCs were successfully isolated. CD44 and CD90 were highly expressed in ACSCs, while CD31, CD34 and CD45 were not expressed in ACSCs. Seven days after culture, single ACSC formed single cell colony containing more than 32 cells. ACSCs demonstrated a high osteogenesis, adipogenesis and chondrogenesis potential. Conclusion ACSCs are present in rat articular cartilages, and ACSCs have typical characteristics of stem cells.

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