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    Monographic report (Pediatric research)
    Effects of Epstein-Barr virus on apoptosis of cord blood monocytes-derived dendritic cells
    JIN Ying-ying, WANG Xi, CHEN Tong-xin
    2010, 30 (2):  121. 
    Abstract ( 1949 )   PDF (5061KB) ( 1281 )  

    Objective To investigate the effects of Epstein-Barr virus (EBV) infection on apoptosis of cord blood monocytes-derived dendritic cells (DC) in neonates. Methods Cord blood monocytes were induced into DC by 50 ng/mL recombinant human granulocyte macrophage colony-stimulating factor (rhGM-CSF) and 10 ng/mL recombinant human interleukin-4 (rhIL-4), and DC were divided into three groups: ①4G group: rhGM-CSF and rhIL-4 were added separately on the day of cell separation; ②4G+0d EBV group: rhGM-CSF, rhIL-4 and B95.8 cell supernatants were added simultaneously on the day of cell separation; ③4G+5d EBV group: hGM-CSF and rhIL-4 were added on the day of cell separation, and B95.8 cell supernatants were added on the 5th day. Percents of DC apoptosis were measured using Annexin V-FITC and PI staining by flow cytometry. The expression of X-linked inhibitor of apoptosis protein (XIAP) was detected by Western Blot. Results Percent of DC apoptosis in 4G+0d EBV group was significantly higher than that in 4G group on the 6th to 14th day (P<0.05), and percent of DC apoptosis in 4G+5d EBV group was significantly higher than that in 4G group on the 7th to 14th day (P<0.05). The effects of EBV infection on percent of DC apoptosis had relations with infection time points. The expression of XIAP in DC decreased significantly after EBV infection. Conclusion EBV promotes apoptosis of cord blood monocytes-derived DC, which is associated with the differentiation and maturation status of DC.

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    Relationship between expression of active form of caspase-3 and cell cycle in Fas-mediated apoptosis of MML-1 cells
    LIN Qian, WU Wei-lan, WEI Min-jiang, et al
    2010, 30 (2):  125. 
    Abstract ( 1881 )   PDF (5931KB) ( 1301 )  

    Objective To investigate the relationship between expression of active form of caspase-3 and cell cycle in Fas-mediated apoptosis of B lymphocytoma cell line MML-1. Methods MML-1 cells were incubated with agonistic anti-Fas antibody for different time, and cell apoptosis was induced. Cell apoptotic rates were analysed by flow cytometry, and sensitivity of MML-1 cells to apoptosis was determined. The expression of active form of caspase-3 was analysed by double staining with PI-Triton X and FITC-active caspase-3. Cyclin A, B1 and E were selected as cell cycle markers for S, G2/M and G1 phase of MML-1 cells, and the expression of active form of caspase-3 was detected by flow cytometry. Results The cell apoptotic rate reached 56% after induction by Fas for 6 h. After induction by Fas for 4 h, the active form of caspase-3 was mainly expressed in cells of G1 phase, while rarely in cells of S and G2/M phase. Cells with negative cyclin A and B1 and positive cyclin E expressed active form of caspase-3. Conclusion The expression of active form of caspase-3 in MML-1 cells mediated by Fas might be cell cycle dependent. Cells entering into late G1 and early S phase first express active form of caspase-3, and their sensitivity to Fas-mediated apoptosis is the highest.

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    Association of single nucleotide polymorphisms in RANTES and Eotaxin-3 genes with childhood asthma
    HUO Jing, LIU Quan-hua, HUA Li, et al
    2010, 30 (2):  129. 
    Abstract ( 1880 )   PDF (2853KB) ( 1256 )  

    Objective To investigate the association of single nucleotide polymorphisms (SNPs) in regulated upon activation normal T cell expressed and secreted (RANTES) gene C-28G (RANTES C-28G), RANTES A-403G and Eotaxin-3 gene C+77T (Eotaxin-3 C+77T) with asthma in Han ethnic children. Methods The buccal mucosa swabs of 192 Han ethnic children with asthma (asthma group) were collected,  and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was employed to detect the SNP loci of RANTES A-403G, C-28G and Eotaxin-3 C+77T. Besides, another 192 healthy subjects (aged 18 to 22 years) without sibship with those in asthma group were served as controls. Genotype and genotypic distribution between these two groups were analysed. Results There was no significant differences in genotype and genotypic distribution of SNP loci of RANTES A-403G and RANTES C-28G between asthma group and control group (P>0.05), while there were significant differences in genotypic distribution of Eotaxin-3 C+77T between these two groups. The frequency of Eotaxin-3 C+77T T/T genotype in asthma group was significantly higher than that in control group (32.3% vs 12.5%, OR=3.44, P=0.000). Conclusion Eotaxin-3 C+77T may be the asthma susceptible SNP loci for Han ethnic children, and Eotaxin-3 C+77T T/T is significantly related with the development of childhood asthma.

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    Clinical analysis of apnea after operation for retinopathy of prematurity
    ZHAO Dong-ying, ZHANG Yong-jun, HE Zhen-juan, et al
    2010, 30 (2):  132. 
    Abstract ( 1647 )   PDF (3500KB) ( 1334 )  

    Objective To retrospectively explore the incidence and causes of apnea after operation for retinopathy of prematurity (ROP). Methods The clinical data of 17 premature infants with operation for ROP (ROP group) and the other 23 premature infants without ROP (control group) were collected. The occurrence of apnea (time of onset and remission and original diseases) was recorded after operation in ROP group and at adjusted 37 weeks of gestational age in control group. Follow-up was conducted for 4 weeks, and the prevalences of apnea were compared between these two groups. Results There was significant difference in prevalences of apnea between ROP group and control group (52.9% vs 21.7%, P<0.05). In ROP group, time of occurrence of apnea was (38±40) h after operation, and time of remission was (126±145) h after onset. Among the infants with apnea, there were 3 cases of pneumonia (33.3%), 1 case of hypoglycemia (11.1%) and 5 cases with unexplained causes (55.5%). Conclusion Infants after operation for ROP are more prone to apnea, and pediatricians and ophthalmologists are required to collaborate in the perioperative care.

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    Research on population pharmacokinetics of Lamotrigine in children with epilepsy
    HE Da-ke, WANG Li, QIN Jiong, et al
    2010, 30 (2):  135. 
    Abstract ( 2002 )   PDF (5444KB) ( 1452 )  

    Objective To establish the population pharmacokinetics(PPK)model of Lamotrigine (LTG) in children with epilepsy in China for promoting individualized dosage regimen. Methods The sparse data of LTG serum concentrations from 60 pediatric patients with epilepsy were collected. One hundred and fourteen serum concentration points were divided into LTG+valproic acid (VPA) group (n=56), LTG+ enzymatic inducer (EI) group (n=26), LTG+EI+VPA group (n=16) and single LTG group (n=16). The serum drug concentrations were the clinical routinely tested steady state concentrations. The LTG PPK parameters were calculated using the non-parametric expectation maximization (NPEM) Program of USC*PACK software, and then a PPK model was established. Based on this model, LTG serum concentrations were predicted with Bayesian fitting program of USC*PACK software. Mean prediction error (MPE) and mean squared prediction error (MSPE) were calculated to evaluate the accuracy and precision of the concentration prediction and to valid the PPK model. Results The greatest likelihood was -192.87. Optimum PPK parameters were: Ka=(1.97 1.66) h-1; Vs=(1.07±0.89) L/kg; Kel=(0.05±0.05) h-1. The linear regression function YOBS=-0.09+1.05 YPRED (R2=0.98, P<0.001), and determination of coefficient was 0.98. MPE was -0.16  g/mL, and MSPE was 0.28 (μg/mL)2. Conclusion A PPK model of LTG in children with epilepsy in China can be successfully established using the USC*PACK software, based on which LTG serum concentrations can be predicted accurately with a Bayesian approach.

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    Clinical analysis of childhood epilepsy with occipital paroxysms
    GE Xin, WU Jie
    2010, 30 (2):  140. 
    Abstract ( 2033 )   PDF (3662KB) ( 1404 )  

    Objective To analyse the differences between idiopathic and symptomatic childhood epilepsy with occipital paroxysms. Methods The medical records of 38 patients with interictal spikes and/or sharp waves over occipital areas, posterior temporal-parietal areas, or both were retrospectively analysed. Epidemiological data, ictal symptoms, electroencephalogram, neuroimaging studies, treatment response and prognosis were compared using chi-square test or Mann-Whitney U test. Results Four patients of  idiopathic group had positive family history related to nervous system. Seizures were more frequent in symptomatic group than in idiopathic group. Both groups showed autonomic symptoms, while ictal vomiting and deviation of eyes were more common in the presence of Panayiotopoulos type. The prognosis of idiopathic group was much better than that of symptomatic group. Conclusion Panayiotopoulos syndrome is more common. Neurologic deficits and electroencephalographic characteristics are major factors to predict seizure remission in children with occipital lobe epilepsy. Idiopathic childhood epilepsy with occipital paroxysms may result in better prognosis.

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    Clinical significance of food allergen-specific IgG detection
    CHEN Yi, CHEN Tong-xin, ZHU Ya-zhong
    2010, 30 (2):  143. 
    Abstract ( 3913 )   PDF (3798KB) ( 1520 )  

    Objective To explore the clinical significance of detection of 14 kinds of food allergen-specific IgG. Methods Fourteen kinds of food allergen-specific IgG were detected by ELISA method in 211 patients with allergic diseases, and IgG positive rates of various foods were compared among patients with different sex, age and allergic diseases. Results Positive food allergen-specific IgG was detected in 193 (91.4%) patients. Among 14 kinds of foods, the positive rate of food allergen-specific IgG was the highest for eggs (73.9%), and milk came the second. However, no elevated food allergen-specific IgG was observed for chicken and meat. Milk was the most common sensitizers for 0-12 month-old patients, and egg was the first cause for the other age groups. There were significant differences in the positive rates of food allergen-specific IgG for milk among different age groups (P<0.01). There was no significant difference in the positive rate of food allergen-specific IgG between males and females (P>0.05). The positive rate of food allergen-specific IgG in patients with eczema was the highest (96.4%), and the lowest (83.3%) was found in those with chronic diarrhea, while there was no significant difference among different diseases (P>0.05). The positive rate of food allergen-specific IgG for milk differed significantly among different diseases (P<0.01). Positive food allergen-specific IgG was detected in 12 kinds of food (except for chicken and meat) for patients with allergic purpura. Conclusion Food intolerance is a complex allergy. The food allergen-specific IgG detection is of great importance as reference for etiologic diagnosis of allergic diseases.

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    Clinical features and electrophysiology analysis of 13 cases of infantile spinal muscular atrophy
    SHEN Ying, ZHANG Jing-min, ZHOU Min-jie, et al
    2010, 30 (2):  147. 
    Abstract ( 2128 )   PDF (4789KB) ( 1379 )  

    Objective To investigate the clinical and electrophysiology features of infantile spinal muscular atrophy, and explore the clinical significance of genetic diagnosis. Methods The clinical data of 13 infants suffering from infantile spinal muscular atrophy were analysed. The serum creatine phosphokinase was examined, and nerve conduction velocity was tested in median nerve, ulnar nerve, tibial nerve and peroneal nerve. The parameters such as distal motor latency, motor nerve conduction velocity and amplitude of compound motor active potential were analysed. Electromyography was performed in no less than four muscles, and the insertion potential, spontaneous potential and motor unit action potential were observed. Deletion of exon 7 in SMN1 gene was detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results All these infants were characterized by progressive flaccid paralysis in limbs. In all cases, amplitude of muscle response was significantly decreased, with prolonged distal latency and slowed conduction velocity. Electromyography demonstrated motoneuron degeneration. Deletion of exon 7 in SMN1 gene was detected in all 13 infants. Conclusion There are unique clinical and electrophysiology features for infantile spinal muscular atrophy, and electromyography may play an important role in the diagnosis. Prenatal genetic diagnosis may help to avoid the birth of this kind of infants.

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    Analysis of professional situation and talent drain of pediatric residents
    LU Mei-hua
    2010, 30 (2):  151. 
    Abstract ( 1967 )   PDF (3228KB) ( 1347 )  

    Objective To investigate the professional situation and talent drain of pediatric residents, and explore the coping strategies. Methods One hundred and twenty pediatric residents and 6 administrative personnels were selected from three hospitals featured in pediatrics. Questionaires were administered to pediatric residents, and interviews were conducted with administrative personnels. Results Only 28.85% pediatric residents were satisfied with present professional situation. The talent drain of pediatric residents was severe, and the major causes were hard work, low incomes and unfavourable career development prospects. Conclusion A considerate number of pediatric residents are not satisfied with the professional situation, and the talent drain is a common occurrence. Government should attach great importance to pediatric medical education and increase health care investment. Besides, the view of value of pediatric residents should be improved and a harmonious medical environment should be established.

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    Monographic report (Gynecologic research)
    Effects of preeclampsia serum on activity and invasive ability of cultured cytotrophoblasts of first trimester of pregnancy
    YAN Shi-lan, TENG Yin-cheng, JIANG Rong-zhen
    2010, 30 (2):  154. 
    Abstract ( 1960 )   PDF (5223KB) ( 1411 )  

    Objective To investigate the effects of preeclampsia serum on activity and invasive ability of cultured cytotrophoblasts of first trimester of pregnancy. Methods Cytotrophoblasts of normal 6 to 8-week pregnancy were cultured by tissue explants adherent method, and were incubated with serum of women with normal pregnancy (normal group) and preeclampsia (preeclampsia group), respectively for 24 h. The activity of cytotrophoblasts was examined by CCK-8, invasive ability was determined by Transwell invasion assay, and expression of MMP-2, MMP-9 and PAI-1 mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR). Results The activity and invasive ability of cytotrophoblasts in preeclampsia group were lower than those in normal group (P<0.05, P<0.01). Compared with normal group, the expression of MMP-2 and MMP-9 mRNA of cytotrophoblasts was significantly lower (P<0.01), and the expression of PAI-1 mRNA was significantly higher (P<0.01). In both groups, the expression of MMP-2 mRNA was negatively related to that of PAI-1 mRNA (r=-0.985, P<0.01; r=-0.933, P<0.05), while there was no correlationship between the expression of MMP-9 mRNA and that of PAI-1 mRNA. Conclusion The preeclampsia serum may affect the invasive ability of cytotrophoblasts by regulating the expression of MMP-2, MMP-9 and PAI-1.

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    Inhibition and radiosensitization of Topotecan, Cisplatin and Taxol on cervical cancer cell line HeLa
    ZHANG Wen-ying, XUE Yue-zhen, TAO Min-fang
    2010, 30 (2):  158. 
    Abstract ( 1832 )   PDF (4313KB) ( 1542 )  

    Objective To investigate the inhibition and radiosensitization effects of Topotecan (TPT) on cervical cancer cell line HeLa, and make comparison with Cisplatin (DDP) and Taxol (TAX). Methods The effects of TPT, DDP and TAX on proliferation of cervical cancer cell line HeLa were evaluated by MTT assay. The radiosensitization effects of TPT, DDP and TAX on HeLa cells were detected by clone formation assay, and the sensitization enhancement ratios of TPT, DDP and TAX were calculated. Results The IC50 of TPT at different reaction time (24 h, 48 h and 72 h) were 8.0 μg/mL, 2.6 μg/mL and 0.8 μg/mL, respectively, those of DDP were 2.4 μg/mL, 0.7 μg/mL and 0.1 μg/mL, respectively, and those of TAX were 0.3 μg/mL, 0.1 μg/mL and 0.0 μg/mL, respectively. The apoptosis rates of tumor cells treated by radiosensitization were significantly higher than those of tumor cells treated by single radiotherapy or single chemotherapy (P<0.05). The sensitization enhancement ratios of TPT at 24 h and 48 h were 1.167 and 1.344, respectively, those of DDP were 1.314 and 1.538, respectively, and those of TAX were 1.076 and 1.316, respectively. Conclusion TPT, DDP and TAX have significant inhibition effects on cervical cancer cell line HeLa, which are time-dependant and dose-dependant. Besides, TPT, DDP and TAX have radiosensitizing effects, with DDP being the strongest and TAX being the weakest.

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    Impact of bilateral ovariectomy on climacteric symptoms in post-menopausal women
    TAO Min-fang, ZHU Jie-ping, TENG Yin-cheng
    2010, 30 (2):  162. 
    Abstract ( 3208 )   PDF (3123KB) ( 1460 )  

    Objective To evaluate the impact of bilateral ovariectomy in post-menopausal women by comparison of climacteric symptoms before and after operation. Methods Post-menopausal women who received hysterectomy with concurrent bilateral ovariectomy on benign uterine indications were recruited. Climacteric symptoms before and after operation were accessed by a questionaire including modified Kupperman Index. Comparison was made after grouping the patients into early menopause group, whose menopausal period was no more than 5 years, and late menopause group, whose menopausal period was more than 5 years. Results Among a total of 95 individuals, 8 had normal, 46 had mild, 34 had moderate, and the other 7 had severe Kupperman Index scores before operation, while 5 had normal, 28 had mild, 48 had moderate, and the other 14 had severe Kupperman Index scores after operation, with more moderate and severe climacteric symptoms after operation than before operation (P<0.05). Women of early menopause group had higher total Kupperman Index scores and symptom scores of sweating, insomnia, depression, fatigue and headache than those of late menopause group before operation (P<0.05). Women of early menopause group had significantly higher Kupperman Index scores after operation than before operation (23.93±7.53 vs 17.63±8.07, P<0.05). However, there was no significant difference in total Kupperman Index scores in women of late menopause group before and after operation. Conclusion Bilateral ovariectomy may affect the climactic symptoms in post-menopausal women, especially in early menopausal period.

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    Relationship between pelvic floor dysfunction and serum relaxin H2 and expression of vaginal wall relaxin receptor LGR7 mRNA in late pregnant women
    JIANG Li, WU Qing-kai, LUO Lai-min, et al
    2010, 30 (2):  165. 
    Abstract ( 1625 )   PDF (3522KB) ( 1387 )  

    Objective To investigate the relationship between pelvic floor dysfunction and serum relaxin H2 and expression of vaginal wall relaxin receptor LGR7 mRNA in late pregnant women. Methods Before the beginning of delivery, all women were evaluated by pelvic organ prolapse quantitation (POP-Q) scoring. Twelve women with stress urinary incontinence (SUI) and stage Ⅱ prolapse of anterior vaginal wall were selected as patient group, and another 24 women without SUI and prolapse of pelvic floor were served as control group. Serum relaxin H2 was determined by ELISA. Vaginal wall tissues were taken after vaginal delivery, and the expression of relaxin receptor LGR7 mRNA was detected by RT-PCR. Results The serum level of relaxin H2 and expression of LGR7 mRNA of vaginal tissues in patient group were significantly higher than those in control group (P<0.01, P<0.05). Conclusion The increased level of serum relaxin and expression of vaginal wall relaxin receptor may correlate with the pelvic floor dysfunction in late pregnant women.

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    Effects of ethinylestradiol and cyproterone acetate on endocrine metabolism in non-obese women with polycystic ovary syndrome
    ZHU Jie-ping, TENG Yin-cheng, TAO Min-fang
    2010, 30 (2):  168. 
    Abstract ( 2007 )   PDF (4318KB) ( 1334 )  

    Objective To investigate the effects of ethinylestradiol and cyproterone acetate tablets (Diane-35) on endocrine metabolism in non-obese women with polycystic ovary syndrome(PCOS). Methods Fifty-three non-obese patients with PCOS were assigned to the 6-month treatment with Diane-35. Body mass index (BMI), waist to hip ratio (WHR), F-G score and acne score were recorded before treatment and at the end of the third and sixth cycle after treatment. Serum total testosterone, follicle stimulating hormone (FSH), luteinizing hormone(LH), LH/FSH, fast plasma glucose, fast insulin and insulin resistance index (HOMA-IR) were obtained at early follicular phase and at the end of the third and sixth month. Results There was no significant change in BMI and WHR after treatment. Compared with those before treatment, F-G score, acne score, serum total testosterone, LH and LH/FSH were significantly decreased at two time points after treatment (P<0.05). BMI and HOMA-IR in patients with insulin resistance were significantly higher at the end of the sixth cycle after treatment than those before treatment (P<0.05). Conclusion Diane-35 can improve hyperandrogenism in non-obese patients with PCOS. However, Diane-35 may aggravate insulin resistance in those with insulin resistance.

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    Diagnostic value of Reid Scores under colposcopy in cervical human papillomavirus infection
    LI Ping, TAO Min-fang
    2010, 30 (2):  171. 
    Abstract ( 1915 )   PDF (3288KB) ( 1320 )  

    Objective To explore the diagnostic value of Reid Scores under colposcopy in cervical human papillomavirus (HPV) infection. Methods Cervical biopsy under colposcopy was carried out for cervical diseases in 528 patients.With pathological findings as criteria, the value of Reid Scores under colposcopy in diagnosis of HPV infection was analysed. Results The coincidence of Reid Scores under colposcopy in diagnosis of HPV infection and cervical intra-epithelial neoplasia (CIN) was 52.66%, with the sensitivity of 51.05% in diagnosis of lowgrade cervical diseases (HPV infection and CINⅠ) and that of 66.67% in diagnosis of high-grade cervical diseases (CINⅡand CINⅢ). For the patients with diagnosis of HPV infection examined by pathology, the scores of items of boundary, color and vessels were positively related to the total Reid Score, while the scores of items of boundary and color were not related to that of vessels. Conclusion There may exist clinical significance in diagnosis of HPV infection by colposcopy in combination with RCI Scores.

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    Original article (Basic research)
    Long-term effects of oxypurinol on oxidative modification of actin and myocardial contractility in mice with heart failure
    TAN Zhen, ZHANG Zhi, DAI Qiu-yan, et al
    2010, 30 (2):  174. 
    Abstract ( 1726 )   PDF (7290KB) ( 1259 )  

    Objective To investigate the long-term effects of xanthine oxidase inhibitor, oxypurinol on myocardial contractility of post-ischemic heart failure in mice, and explore the underlying mechanism. Methods One hundred and twenty SV120 mice were randomly assigned into myocardial infarction control group, sham operation group and Oxy treatment group. Post-ischemic heart failure were induced by left anterior descending coronary artery ligation in myocardial infarction control group and Oxy treatment group, and mice in Oxy treatment group and sham operation group were orally administered with 0.5 mmol/L Oxy each day. Nine to eleven months after treatment, echocardiography was performed in all groups. Trabeculae from the right ventricle of mice were dissected for assessment of changes in excitation-contraction coupling. Sarcomere length was measured by laser diffraction. Intracellular free Ca2+ concentration ([Ca2+]i) was detected with fluorescent dye Fura-2, which was microinjected iontophoretically into cells. Steady-state force-[Ca2+]i was achieved by addition of ryanodine and increasing the stimulus frequency to induce tetanization, and the relationship between myocardial contractility and intracellular Ca2+ transients was analysed. Besides, Western blotting was performed to determine the oxidation of myofilament proteins. Results Long-term oral administration of oxypurinol significantly improved myocardial contraction function and reduced ventricular wall thickness. Programming of excitation-contraction coupling was significantly improved, and maximal Ca2+ activated force (Fmax) in steady-state was also significantly increased. Western blotting revealed the oxidative modification of actin in mice of Oxy treatment group was significantly inhibited compared with that of myocardial infarction control group. Conclusion Long-term treatment with Oxy improves the cardiac contraction function and boosts the cardiac force dramatically in post-ischemia heart failure. The increase in contraction is the result of increased myofilament Ca2+ responsiveness. Thus, antioxidant oxypurinol, by preventing oxidative damage to contractile proteins, can augment contraction with little changes in [Ca2+]i, represents new class of inotropic agents with advantages of reducing Ca2+ overload, and offers new promises in management of heart failure in the future.

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    Identification of cardiomyocytes differentiated from mesenchymal stem cells and study on expression of Nesprin protein
    YANG Wen-gang, XUE Song, WANG Zheng, et al
    2010, 30 (2):  180. 
    Abstract ( 2206 )   PDF (9604KB) ( 1341 )  

    Objective To induce and identify the differentiation of rat bone marrow mesenchymal stem cells (MSCs) into cardiomyocytes in vitro, and observe the expression of Nesprin protein during the differentiation. Methods Rat MSCs were isolated and purified by Ficoll density gradient centrifugation, and adhered for serial subcultivation. Surface-associated antigens of MSCs of the second passage were dedected by flow cytometry. MSCs of the second passage were induced by 10 μmol/L 5-azacytidine (5-Aza) to differentiate into cardiomyocytes, and the morphological changes were observed. The expression of Desmin, α-sarcomeric actin and cardiac Troponin I (cTnI) mRNA and protein was detected by RT-PCR, immunocytochemistry and immunofluorescence staining, and the expression of Nesprin protein was detected by Western blotting. Results The morphology of MSCs induced by 5-Aza was bigger and longer, and the nuclei became bigger, exhibiting more consistent patterns. The expression of Desmin, α-sarcomeric actin and cTnI mRNA and protein of MSCs was positive. Immunofluorescence staining revealed that Nesprin protein positioned in the nuclear membrane, and Western blotting detection demonstrated that the expression of Nesprin protein significantly increased after differentiation (P<0.05). Conclusion MSCs may be successfully induced to differentiate into cardiomyocytes. The expression of Nesprin protein in the differentiated MSCs may significantly increase, indicating Nesprin may play a role in the differentiation from MSCs to cardiomyocytes.

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    Effect of probiotics on intestinal flora disturbance and bacterial translocation in mice with spontaneous colitis
    WANG Jian-qiang, DING Zai-xian, ZHANG Min, et al
    2010, 30 (2):  186. 
    Abstract ( 2495 )   PDF (5987KB) ( 2192 )  

    Objective To investigate the effect of Lactobacillus plantarum (LP) on intestinal flora and bacterial translocation in mice with spontaneous inflammatory bowel disease (IBD). Methods Interleukin 10 knockout mice (IL-10-/-) were used as models of IBD. Eight-week old female mice were randomized to control group, IL-10-/- group and IL-10-/-+LP group. IL-10-/-+LP group received 0.5 mL LP (1.0×109 CFU/mL) per day for 4 weeks, and the other groups received 0.5 mL Ringer buffer. Intestinal flora including Bifidobacteria, Lactobacilli, Enterobacteriaceae and Clostridium perfringens in the feces and bacterial translocation in mesenteric lymph nodes and spleens were detected. Results The contents of Bifidobacteria and Lactobacilli significantly decreased in the intestine of IL-10-/- mice, while those of Enterobacteriaceae and Clostridium perfringens significantly increased, and the bacterial translocation significantly increased. Four weeks after LP treatment, the disturbed intestinal flora was restored, and the bacterial translocation decreased. Conclusion LP administration can modulate the imbalance of intestinal flora and decrease the bacterial translocation, thus enhance intestinal barrier function in mice with IBD.

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    Role of calcitonin gene-related peptide and IP3 pathway in ischemic preconditioning in isolated perfused hearts of rats with type 1 diabetes mellitus
    WANG Shi-ting, GUO Zhu-ying, XU Mang-hua, et al
    2010, 30 (2):  191. 
    Abstract ( 2030 )   PDF (5835KB) ( 1511 )  

    Objective To investigate the role of calcitonin gene-related peptide (CGRP) and IP3 signal pathway in ischemic preconditioning (IPC) in the isolated perfused hearts of rats with type 1 diabetes mellitus. Methods Type 1 diabetes mellitus rat models were established in 80 SD rats, and were randomly divided into 4 week (D-4w) group and 8 week (D-8w) group. These two groups were randomly subdivided into model control (D-Cont) group, type 1 diabetes mellitus ischemia-reperfusion (IR) group, IPC group, CGRP (IPC+CGRP) group and IP3 inhibitor wortmanin (IPC+WMN) group. Another 16 rats were served as normal control (N-Cont) group. In vitro perfusion models of isolated hearts were established by Langendorff methods, and CGRP or wortmanin (WMN)were administered during perfusion. The left ventricle function of isolated heart in each group was monitored by multichannel biosignal analysis system, and coronary artery flow was recorded. The serum CGRP levels were detected by ELISA. The activity of lactate dehydrogenase (LDH) and creatine kinase (CK) in effluent of coronary artery was detected by biochemical method. The size of myocardial infarction was determined by NBT staining, and apoptosis of cadiocytes was detected by TUNEL method. Results Compared with N-Cont group, the CGRP level in serum of rats with type 1 diabetes mellitus decreased with time, the basic left ventricle function decreased, while the activity of LDH and CK in effluent of coronary artery, size of myocardial infarction and cardiomyocyte apoptosis index increased (P<0.05). Compared with N-Cont group, the left ventricle function was significantly lower in IR group, and more severe myocardial damage was observed. IPC improved myocardial damage of D-4w IR group, while had no protection on D-8w IR group. Compared with IPC group, the left ventricle function was significantly improved in IPC+CGRP group. IPC+WMN blocked the myocardial protection of D-4w group from IPC. Conclusion CGRP and IP3 signal pathway are involved in the protection provided by IPC in isolated hearts of rats with type 1 diabetes mellitus.

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    Long-term effect of ketamine on learning and memory function of postnatal rats
    SHEN Bo-xiong, QU Xing-qian, CHENG Zhi-jun
    2010, 30 (2):  196. 
    Abstract ( 1793 )   PDF (5304KB) ( 1473 )  

    Objective To investigate the effect of ketamine on learning and memory function of adult rats after injection of ketamine at early development stage. Methods Twelve rats born in a week were randomly divided into ketamine anesthesia group (Ket group) and normal saline control group (Ns group). Rats in Ket group were intraperitoneally injected with 50 mg/kg ketamine, and those in Ns group were administrated with same amount of normal saline. After eight weeks, Morris water maze test was adopted to explore the latency in place navigation and spatial probe ability. The sections of hippocampus were obtained, and immunohistochemical staining was employed to detect the expression of Bax and Bcl-2. Results In Morris water maze test, the latency in place navigation of Ket group was significantly longer than that of Ns group (P<0.05), and the spatial probe ability significantly decreased (P<0.05). It was revealed by immunohistochemical staining that the expression of Bax in Ket group was significantly higher than that in Ns group, while there was no significant difference in the expression of Bcl-2 between these two groups. Conclusion Application of ketamine in neonatal rats may result in learning and memory impairment in adulthood, and the underlying mechanism may correlate to the apoptosis of neuron in hippocampus region.

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    Changes in proliferative capability of remnant breast tumor after ablation with high intensity focused ultrasound
    NIU Ling-chuan, ZOU Wen-bing, ZHANG Lian, et al
    2010, 30 (2):  200. 
    Abstract ( 1933 )   PDF (8033KB) ( 1196 )  

    Objective To observe the changes in proliferative capability of remnant tumors implanted in rabbit breast after ablation with high intensity focused ultrasound (HIFU). Methods Rabbit implanted models were established by implanting VX2 tumor mass into the breast of New Zealand rabbits. HIFU was applied to the tumors, and the remnant tumors were harvested by controlling temperature around probe of tumors. Fifty-six rabbits were randomly divided into control group(n=10) and HIFU group(n=40). The expression of proliferating cell nuclear antigen (PCNA) was detected by immunohistochemistry at different time points after ablation, and the survival time and organ metastasis status were observed after ablation. Results After ablation in HIFU group, the positive expression of PCNA was decreased shortly in remnant tumors, and restored to the level before ablation 21 days after treatment. The survival time in HIFU group was much longer than that in control group, and the time of lung and abdominal viscera metastasis in HIFU group appeared significantly later than that in control group (P<0.05). Conclusion Though HIFU ablation can not completely eradicate tumor after one treatment, the growth and metastasis of remnant tumors can be effectively inhibited for a short time, and HIFU ablation can prolong the survival time.

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    Preliminary study on AmpC β-lactamase and related drug-resistant mechanism of pathogenic bacteria in blood stream
    HOU Wei-wei, JIANG Yan-qun
    2010, 30 (2):  204. 
    Abstract ( 1562 )   PDF (4265KB) ( 1271 )  

    Objective To investigate the characteristics of strains of AmpC β-lactamase (AmpC enzyme) production in pathogenic bacteria in blood stream and clinical presentations of the cases, and study the related ampC and ampD genes. Methods One hundred and eighty-one strains of gram negative bacillus in blood stream were collected, Cefoxitin screening test and three-dimensional test were performed for screening of strains of AmpC enzyme, production and those of AmpC enzyme hyperproduction retrospective analysis was condected in the strains with positive results. ampC and ampD gene PCR amplification, sequencing and sequence analysis of positive strains were performed, and gene homology of ampC positive strains was analysed by Rep-PCR. Results Among 181 strains in blood stream, strains of AmpC enzyme production were detected in 39 isolates by Cefoxitin screening test, with the detection rate of 21.5% (39/181). The detection rate of strains of AmpC enzyme hyperproduction by three-dimensional test was 43.6% (17/39). PCR revealed that the positive rates for ampC and ampD genes were 41% (16/39) and 56.4% (22/39), respectively. The ampC gene sequencing of 16 positive strains indicated that the homology was 98% to 100% by comparison with the GenBank, while the ampD gene sequencing of 2 strains of Enterobacter cloacae demonstrated that the suspected gene mutations existed in the carboxy-terminal of ampD gene. Conclusion The prevalence of drug-resistant pathogenic bacteria in blood stream in this study is due to nosocomial infection. The mutation of ampC gene is rare in the pathogenic bacteria in blood stream with production of AmpC enzyme, while the rate of gene mutation in Enterobacter cloacae is higher, and the deletion and amino acid substitutions in the carboxy-terminal of ampD is highly relevant to the depressed expression of AmpC enzyme.

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    Original article (Clinical research)
    Value of GPC3 and PEG10 mRNA in peripheral blood in diagnosis of metastasis in hepatocellular carcinoma
    ZHANG Jie, WANG Zhi-qing, YU Jia-ping, et al
    2010, 30 (2):  208. 
    Abstract ( 1752 )   PDF (6029KB) ( 1392 )  

    Objective To explore the value of glypican-3 (GPC-3) mRNA and paternally expressed 10 (PEG10) mRNA in peripheral blood in diagnosis of metastasis in hepatocellular carcinoma (HCC). Methods With SYBR Green I as fluorescence signal, real-time fluorescent quantitative reverse transcription-polymerase chain reaction was employed to detect the expression of GPC-3 mRNA and PEG10 mRNA in peripheral blood from patients with HCC with metastasis (n=8), HCC without metastasis (n=12) and hepatic cirrhosis (n=11), and receiver operator characteristics curve (ROC) and specific parameters were adopted to analyse their value in predictive and exclusive diagnosis. Results The expression of GPC-3 mRNA and PEG10 mRNA in HCC with metastasis was significantly higher than that in HCC without metastasis and in hepatic cirrhosis (P<0.05), while there was no significant difference in the expression of GPC-3 mRNA and PEG10 mRNA between HCC without metastasis and hepatic cirrhosis. In single test, the sensitivities in the differential diagnosis between HCC with metastasis and HCC without metastasis were 66.7% for GPC-3 mRNA and 72.2% for PEG10 mRNA, and the specificities were 91.7% and 91.7%, respectively. The areas under ROC were 0.748 for GPC-3 mRNA and 0.812 for PEG10 mRNA. With two markers in parallel test, the sensitivity, specificity, negative likelihood and diagnostic accuracy were 90.7%, 84.0%, 0.11 and 83.3%, respectively. In serial test, the sensitivity, specificity, positive likelihood and diagnostic accuracy were 60.5%, 98.7%, 45.5 and 73.3%, respectively. Conclusion Detection of GPC-3 mRNA and PEG10 mRNA in peripheral blood may help to predict blood metastasis and extrahepatic metastasis of HCC, and PEG10 mRNA works better than GPC-3 mRNA. The serial test of GPC-3 mRNA and PEG10 mRNA is helpful to the predictive diagnosis of peripheral blood metastasis of HCC.

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    Expression of CD133 mRNA in tissues of gastric cancer and its relationship with clinicopathological features
    ZHANG Peng, WU Ju-gang, WU Hong-biao, et al
    2010, 30 (2):  213. 
    Abstract ( 1746 )   PDF (7567KB) ( 1213 )  

    Objective To investigate the expression of CD133 mRNA in tissues of gastric cancer, and explore its relationship with clinicopathological features. Methods The tissues of gastric cancer and normal tissues adjacent to gastric cancer were obtained from 31 patients. The expression of CD133 mRNA was detected by semi-quantitative RT-PCR, and its relationship with clinicopathological features such as sex, age, tumor diameter, infiltration depth, TNM staging, tumor differentiation, lymph node metastasis and Ki-67 proliferation index was analysed. Results The relative gray scale values of CD133 mRNA in tissues of gastric cancer and normal tissues adjacent to gastric cancer were 0.378 3±0.141 1 and 0.038 1±0.091 9, respectively (P=0.000). The relative gray scale values of CD133 mRNA in tissues of gastric cancer with tumor diameter >5 cm were significantly higher than those with tumor diameter ≤5 cm [(0.439 3±0.148 4) vs (0.334 3±0.121 2)](P=0.041), and those in tissues with lymph node metastasis were significantly higher than those without lymph node metastasis [(0.426 6±0.132 0) vs (0.239 5±0.030 9)](P=0.004). The rate of lymph node metastasis and the number of metastatic lymph nodes were positively related to relative gray scale values of CD133 mRNA(r=0.466, P=0.008; r=0.464, P=0.009). The relative gray scale values of CD133 mRNA in those with low expression of Ki-67 were significantly higher than those with high expression of Ki-67 [(0.436 4±0.139 8) vs (0.316 4±0.117 4)](P=0.02), and expression of Ki-67 were negatively related to relative gray scale values of CD133 mRNA (r=-0.461, P=0.009). Conclusion The expression of CD133 mRNA in tissues of gastric cancer was associated with the rate of lymph node metastasis, number of metastatic lymph nodes and expression of Ki-67, which reflect the status of lymph node metastasis and proliferation of gastric cancer.

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    Study of obstructive sleep apnea-hypopnea syndrome and metabolic syndrome in females
    MA Guan-hua, LI Min, CHEN Ling, et al
    2010, 30 (2):  218. 
    Abstract ( 1862 )   PDF (4394KB) ( 1508 )  

    Objective To analyse the changes of metabolic parameters and condition of metabolic syndrome (MS) in females with obstructive sleep apnea-hypopnea syndrome (OSAHS), and explore the interaction between OSAHS and MS. Methods Sixty females undergoing polysomnography were included, and were divided into simple snorer group (n=19), mild OSAHS group (n=21) and moderate to severe OSAHS group (n=20). Epworth sleepiness scale (ESS) scores and metabolic parameters were compared among groups. Forty-one patients with OSAHS were subdivided into group with MS (MS group, n=15) and group without MS (non-MS group, n=26). ESS scores, apnea hypopnea index (AHI), oxygen desaturation index (ODI) and lowest pulse oxygen saturation (LSpO2) were compared between MS group and non-MS group. Results There was no significant difference in age, body mass index (BMI), total cholesterol (TC), triglyceride (TG), low density cholesterol (LDL), high density cholesterol (HDL), apolipoprotein A-I (apoA-I) and apolipoprotein B(apoB) among simple snorer group, mild OSAHS group and moderate to severe OSAHS group (P>0.05). Correlation analysis revealed ESS was positively related to AHI (r=0.327, P=0.011). ESS scores and proportions of hypertension, fasting plasma glucose (FPG) and MS were significantly higher in moderate to severe OSAHS group than those in simple snorer group (P<0.05). BMI, FPG, TG, proportion of hypertension, ODI and ESS score in MS group were significantly higher than those in non-MS group (P<0.05), while there was no significant difference in AHI and LspO2 between these two groups (P>0.05). Conclusion ESS score may reflect the severity of OSAHS in females. Females with OSAHS matched for age and BMI have no difference in blood fat. With the increase of severity of OSAHS, glycometabolism can be impaired, the prevalences of hypertension and MS increase. MS can exacerbate the severity of OSAHS, indicating that MS and OSAHS interact with each other.

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    Relationship between expression of RANTES and microvascular density in tissues of primary hepatocarcinoma
    XIANG Zhi-gang, YANG Zhu-lin, HUANG Jian-jun
    2010, 30 (2):  222. 
    Abstract ( 1901 )   PDF (5145KB) ( 1331 )  

    Objective To investigate the relationship between the expression of regulated-upon activation normal T expressed and secreted (RANTES) and microvascular density (MVD) in hepatocarcinoma (PHC) tissues, and explore its clinical significance. Methods The samples of PHC tissues and adjacent tissues in 47 patients were collected. The expression of RANTES in tissues was detected by immunohistochemistry, and MVD was calculated. The relationship among the expression of RANTES, MVD and clinicopathological features was analysed. Results The positive expression rate and expression score of RANTES in PHC tissues were significantly higher than those in adjacent tissues (55.32% vs 19.15%, P<0.01; 1.89±1.77 vs 0.77±1.29, P<0.01). MVD in PHC tissues was significantly higher than that in adjacent tissues (67.30±13.68 vs 37.20±10.58, P<0.01). MVD of PHC tissues in patients with metastasis was significantly higher than that in patients without metastasis [(73.50±13.77)/HP vs (64.10±12.68)/HP, P<0.05], while there were no correlations among MVD, expression of RANTES and the other clinicopathological features of PHC. MVD was positively correlated with the expression score of RANTES in PHC tissues (r=0.386, P<0.05). Conclusion RANTES might be closely related to the angiogenesis in PHC.

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    Review
    Canonical Wnt pathway and schizophrenia
    WANG Xue-mei
    2010, 30 (2):  225. 
    Abstract ( 1678 )   PDF (4297KB) ( 1190 )  

    Schizophrenia is one of the most severe psychiatric disorders. Although the etiology is unknown, schizophrenia appears to be a polygenic disorder associated with environmental factors. There are evidences that intracellular signaling plays an important role in the pathogenesis of schizophrenia. Canonical Wnt pathway, which is closely related to the development and function maintenance of central nervous system, have drawn wide attention. Evidences for potential disturbances of Wnt signaling pathways in schizophrenia have accumulated, which are in favour of the hypothesis that Wnt signaling alterations may be important in the pathogenesis of schizophrenia. The research advances in the relationship between canonical Wnt signaling pathway and schizophrenia are reviewed in this paper.

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    Research advance in role of ROS in cardioprotection offered by preconditioning or postconditioning
    XIN Ping
    2010, 30 (2):  229. 
    Abstract ( 1464 )   PDF (4327KB) ( 1243 )  

    Reactive oxygen species (ROS) play an important role in the cardiac ischemic and reperfusion injury. However, accumulating evidences have demonstrated that ROS are pivotal components of redox signaling cascade relevant to cardioprotection induced by either preconditioning or postconditioning. In this review, the definition, source and detection methods of ROS are introduced. Besides, the role of ROS in the cardioprotection offered by preconditioning or postconditioning and the effects of either preconditioning or postconditioning on ROS are summarized.

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    Research advances in effects of general anesthetics on central nervous system of pediatric patients and immature animals
    ZHANG Qing-qing
    2010, 30 (2):  233. 
    Abstract ( 1651 )   PDF (3521KB) ( 1283 )  

    Long-term or even permanent neuronal injury can follow administration of general anesthetics. It has been confirmed that some elderly patients may experience a decline in cognitive function after general anesthesia. However, the effects of general anesthetics on the central nervous system have not been determined yet. Here we summarize the effects of general anesthetics on the central nervous system in pediatric patients and immature animals.

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    Research advances in precancerous lesions of stomach
    ZHANG Ye-li
    2010, 30 (2):  236. 
    Abstract ( 1964 )   PDF (4150KB) ( 1340 )  

    Gastric precancerous lesion refers to epithelial dysplasia (atypical hyperplasia or intraepithelial neoplasias), which is associated with increased risk of gastric cancer. It happens to be controlled by multiple factors and/or polygene such as the H. pylori infection, diet and environment which play an important role in the development of gastric precancerous lesions. This article describes the pathogenesis of gastric precancerous lesions as many scholars have studied some of it.

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    Technique and method
    Synthesis of substrate for angiotensin-converting enzyme and its value in diagnosis of lung cancer
    ZHOU Hai-yan, YAO Li-yun, LI Ying-yi, et al
    2010, 30 (2):  240. 
    Abstract ( 1903 )   PDF (4599KB) ( 1512 )  

    Objective To establish the synthetic method of the substrate for angiotensin-converting enzyme (ACE), and investigate the assay method of serum ACE. Methods The substrate was prepared by the use of N-hydroxysuccinimide ester in peptide synthesis, and serum ACE was determined by spectrophotometric assay using substrate. Results The substrate of furanacryloyl-phenylalanyl-glycyl-glycine with purity of 99.5% was obtained. The Km value for the substrate was 0.25 mmol/L, the normal reference interval for serum ACE activity was 46.7 U/L to 113.3 U/L, and the within-run and between-run coefficient of variation (CVs) ranged from 3.9 to 4.9%. The serum ACE activity in patients with squamous carcinoma and adenocarcinoma of lung were significantly decreased (P<0.05), while that in patients with lung small cell carcinoma was not significantly decreased (P>0.05). Conclusion The synthesized substrate could be reliably used as a marker for diagnosis of different lung cancers in clinics.

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    Spectrophotometric study of Poloxamer in contact lens care solution
    LI Jia-zhong, ZHANG Li, HE Tao, et al
    2010, 30 (2):  244. 
    Abstract ( 1948 )   PDF (3734KB) ( 1465 )  

    Objective To determine the contents of Poloxamer in contact lens care solution by spectrophotometric method, and investigate the effect of the other lubrication composition on the measurement. Methods Five brands of contact lens care solution with hydroxypropyl methyl cellulose (HPMC) were obtained. The interference of HPMC in contact lens care solution was removed by heating and rapid filtering, and the contents of Poloxamer was determined by KI-I2 spectrophotometry. Results For the pre-treated contact lens care solution, the tested wavelength was 500 nm and the development time was 120 min. Favourable linearity was observed when Poloxamer ranged between 0.001 mg/mL and 0.020 mg/mL, with standard curve of Y=0.050 91X+0.017 15 and r2=0.998 2. Precision (n=6) test revealed that RSD was 1.331 4%, and RSD of daytime repeatability experiment was 2.971 3%. The recovery rates of 80%, 100% and 120% samples ranged between 90% to 105%. Conclusion KI-I2 spectrophotometry is a convenient, stable and reliable method, and may serve as a mass control method for nonionic surfactant Poloxamer content in contact lens care solution.

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    Case report
    Two cases report of psoriasis complicated with coronary heart disease
    HU Wei, GU Jun, ZHANG Da-dong
    2010, 30 (2):  247. 
    Abstract ( 1595 )   PDF (2079KB) ( 979 )  

    Recently, the relationship between psoriasis and coronary heart disease (CHD) have been casting close attention. It has been demonstrated that the prevalence of CHD in patients with psoriasis is more than one time higher than that of healthy people. Meanwhile, the prevalences of hypertension, diabetes mellitus, hyperlipoidemia and smoking in patients with psoriasis are higher than those of normal people, and some drugs for treatment of psoriasis may induce CHD. Therefore, the relationship between psoriasis and CHD is still unclear. In this paper, two cases of psoriasis complicated with CHD are reported, and the relationship between psoriasis and CHD is explored.

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