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    Basic research
    Inhibitory effect of sanguinarine on proliferaton and invasion of gastric cancer cells by upregulating m6A methyltransferase 14
    Ming CHEN, Jing ZHANG
    2022, 42 (2):  135-141. 
    doi: 10.3969/j.issn.1674-8115.2022.02.001

    Abstract ( 166 )   HTML ( 1161 )   PDF (2415KB) ( 250 )  
    Objective

    ·To investigate the effect of sanguinarine (SAG) on the proliferation and invasion of gastric cancer cells MGC-803 and AGS, and the relationship between the mechanism and N6-methyladenosine (m6A) methyltransferase 14 (METTL14).

    Methods

    ·After the gastric cancer cell lines (MGC-803 and AGS) were exposed to different concentrations of SAG (0, 10, 20 μmol/L) for 48 h, quantitative PCR and Western blotting analysis were used to detect the effect of SAG on the expression of METTL14. Then, MGC-803 and AGS cells transfected with lentiviruses-mediated small interfering RNA of METTL14 (si-METTL14) or control (si-NC) were treated with 10 μmol/L SAG or PBS for 48 h, and thus the two cell lines were divided into si-METTL14+SAG group, si-NC+SAG group and si-NC+PBS group, respectively. Quantitative PCR and Western blotting were used to verify the expression levels of METTL14 after it was transfected with si-METTL14 in gastric cancer cells. The proliferation level, number of clones formed and invasion potential of the 3 groups in both MGC-803 cells and AGS cells were observed by MTT proliferation assay, cell clone formation test and Transwell invasion assay, respectively. Independent samples t test was used for comparison between two groups of data, and one-way ANOVA was used for comparison between more than two groups of data.

    Results

    ·Compared with the control group (0 μmol/L), 10 μmol/L SAG and 20 μmol/L SAG up-regulated METTL14 mRNA and protein expression levels (P<0.05), showing a certain concentration dependence. Quantitative PCR and Western blotting results confirmed that the expression levels of METTL14 in gastric cancer cells were significantly reduced after transfection of si-METTL14 in both MGC-803 cells and AGS cells. MTT cell proliferation assay showed that the cell proliferation rate of the si-NC+SAG group was significantly lower than that of the si-NC+PBS group in each cell line (P=0.000). The cell clone formation test showed that the number of cell clones of the si-NC+SAG group was significantly smaller than that of the si-NC+PBS group in each cell line (P<0.01). The Transwell invasion assay showed that the cells crossing Matrigel gel in the si-NC+SAG group was significantly less than that in the si-NC+PBS group in each cell line (P<0.01). si-METTL14 could partially reverse the inhibitory effects of SAG on gastric cancer cells.

    Conclusion

    ·SAG can inhibit the proliferation activity, clonal formation and invasion potential of gastric cancer cells, which may be realized by upregulating METTL14 expression level.

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    Application of three-dimensional electron microscopy to morphological study of neurons in brainstem cochlear nucleus
    Jialei ZHOU, Haibin SHENG, Haoyu WANG, Yan LU, Fangfang WANG, Hao WU, Yunfeng HUA
    2022, 42 (2):  142-149. 
    doi: 10.3969/j.issn.1674-8115.2022.02.002

    Abstract ( 105 )   HTML ( 1175 )   PDF (3073KB) ( 186 )  
    Objective

    ·To explore the feasibility of cross-scale neuroanatomy and connectomics of mouse cochlear nucleus (CN) by using a new three-dimensional (3D) electron microscopic imaging method.

    Methods

    ·The intact CN was obtained after the brain tissue of adult CBA/Ca mice (2 months old) was fixed and dissected, stained with heavy metals (reducing osmium amplification method) , dehydrated with gradient ethanol and anhydrous acetone and embedded with low viscosity resin. The CN tissue was imaged by X-ray microscopy and 3D reconstruction, and its staining quality was evaluated. Then targeted subdivision was located according to the specific distribution of auditory nerve fibers. The low-resolution pre-scan of the CN tissue was performed by scanning electron microscopy, and compared with X-ray microscopic dataset. After the target location was determined, the volume of interest was mapped by serial block-face scanning electron microscopy (SBEM) for 3D reconstruction, and the ultrastructures of bushy cells and the synapses projecting on its surface were tracked, annotated and reconstructed.

    Results

    ·3D electron microscopic samples of intact CN of CBA/Ca mice were successfully prepared. The 3D structure of CN with cell resolution was collected and reconstructed by X-ray microscopic imaging, and the anatomical localization of subregions of CN was achieved. The 3D electron microscopic data of bushy cells in the target area of CN were successfully collected by SBEM. The tracking, labeling and reconstruction of endbulb of Held synapses and other non-auditory synapses on the cell body of target bushy cells were completed. The data of 3D electron microscopy showed that there were 5 endbulb of Held synapses projecting to the surface of the target bushy cell, forming a total of 348 synaptic active zones, while there were 97 synapses from non-auditory nerve.

    Conclusion

    ·It is feasible to prepare 3D electron microscopic samples of adult mouse intact CN by Osmium-based enbloc staining and resin embedding. X-ray microscopic imaging can be used for rapid and accurate localization of subregions and target volume of CN, and the acquired 3D electron microscopic data can be used to study the neuronal morphology and synaptic connections in the CN.

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    Effects of Pcsk9 gene interference on high fat-induced nonalcoholic fatty liver disease with atherosclerosis in rats
    Xiaowen ZHANG, Yi WANG, Chan ZHANG, Di ZHANG, Hang YUN, Di HUANG
    2022, 42 (2):  150-157. 
    doi: 10.3969/j.issn.1674-8115.2022.02.003

    Abstract ( 128 )   HTML ( 1001 )   PDF (2405KB) ( 193 )  
    Objective

    ·To investigate the effects of proprotein convertase subtilisin kexin 9 (Pcsk9) gene knockdown on non-alcoholic fatty liver disease (NAFLD) and atherosclerotic lesions in rats induced by high fat.

    Methods

    ·The SD rat model of NAFLD was established. Rats were randomly divided into 4 groups: control group, model group (high fat), shRNA-negative control (NC) interference model group (high fat+shRNA-NC) and Pcsk9-shRNA interference model group (high fat+ Pcsk9-shRNA). Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect Pcsk9 gene interference efficiency. Fasting serum insulin was determined by radioimmunoassay. Automatic biochemical analyzer was used to detect levels of blood lipid in rats. Hematoxylin-eosin staining (H-E staining) was used to observe the injury of liver tissue and aorta tissue. Apoptosis of liver tissue was detected by TUNEL staining. The levels of interleukin-1β (IL-1β), IL-6 and inducible nitric oxide synthase (iNOS) in peripheral blood were detected by enzyme linked immunosorbent assay (ELISA). The expression of PCSK9, Toll-like receptor 4 (TLR4), nuclear factor κB (NF-κB) P65 and tumor necrosis factor α (TNF-α) were detected by Western blotting.

    Results

    ·Compared with the control group, obesity index and insulin level in the model group were significantly increased (all P=0.000); the apoptosis rate of liver cells was significantly increased (P=0.000); the level of high density lipoprotein cholesterol (HDL-C) was significantly decreased, while the levels of low density lipoprotein cholesterol (LDL-C), total cholesterol (TC) and triacylglycerol (TAG) were significantly increased (all P=0.000); the levels of IL-1β, IL-6 and iNOS were significantly increased (all P=0.000); TLR4, NF-κB P65 protein activation and TNF-α expression were significantly increased (all P=0.000); the liver tissue and aorta tissue were significantly damaged. After interferenceof Pcsk9 gene expression, compared with the model group, obesity index and insulin level in the high fat+Pcsk9-shRNA group were significantly reduced (P=0.007, P=0.000); the apoptosis rate of liver cells was significantly reduced (P=0.000); the level of HDL-C was significantly increased while the levels of LDL-C, TC and TAG were significantly decreased (all P=0.000); the levels of IL-1β, IL-6 and iNOS were significantly decreased (all P=0.000); TLR4, NF-κB P65 protein activation and TNF-α expression were significantly decreased (all P=0.000); the histopathological lesions of liver tissue and aorta tissue were improved.

    Conclusion

    ·Knockdown of Pcsk9 gene can reduce obesity index, insulin level, blood lipid index and inflammatory response in the rats with NAFLD and atherosclerosis, and inhibit the activation of TLR4 and NF-κB P65 protein, thereby improving the injury of liver and aortic tissue in rats.

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    Interventional effects of nicotinamide mononucleotide on metabolism in aging mice
    Guodong DANG, Xinyu HONG, Meiqin CAI
    2022, 42 (2):  158-165. 
    doi: 10.3969/j.issn.1674-8115.2022.02.004

    Abstract ( 155 )   HTML ( 50 )   PDF (1837KB) ( 216 )  
    Objective

    ·To investigate the effects of nicotinamide mononucleotide (NMN) on metabolism in aging mice.

    Methods

    ·Seventy C57BL/6N male mice were randomly divided into 5 groups by using a table of random numbers. They were the control group, the premature aging model group, the aging model group, the intervention group Ⅰand the intervention group Ⅱ. Each group contained 14 mice. Except the control group, D-galactose (D-gal) (150 mg/kg) was subcutaneously injected into the napes of mice in the other 4 groups to establish the aging model of mice. NMN (300 mg/kg) was given to the intervention group Ⅰ and the intervention group Ⅱ by intragastric administration at the same time, and the other groups were given the same amount of distilled water, once a day, for 6 weeks in the premature aging model group and the intervention group Ⅰ, and for 12 weeks in the aging model group and the intervention group Ⅱ. The control group was given the same amount of normal saline and distilled water, once a day, for 6 weeks. Six weeks after modeling, the energy metabolism levels of the mice in the control group, the premature aging model group and the intervention group Ⅰ were detected, including respiratory metabolism, activity level and energy consumption. The organ indexes of thymus, spleen, liver and kidney were calculated. The glucose tolerance and insulin sensitivity were measured. In addition, the content of superoxide dismutase (SOD) and the activities of glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) in the serum and liver tissue were detected. Twelve weeks after modeling, the above indexes were detected in the aging model group and the intervention group Ⅱ.

    Results

    ·Compared with the control group, the thymus index (P=0.035, P=0.000) and renal index (P=0.009, P=0.002) of the model groups were significantly decreased. The O2 consumption (P=0.018, P=0.000), CO2 exhalation (P=0.044, P=0.003), energy consumption (P=0.010, P=0.001) and activity ability (both P=0.000) of the premature aging model group and the aging model group were significantly decreased at night. The insulin sensitivity was significantly reduced (P=0.012, P=0.011). The activities of SOD (P=0.002, P=0.001) and GSH-Px (P=0.001, P=0.011) in serum were significantly decreased and the content of MDA in serum was significantly increased (both P=0.000). The decline of energy metabolism levels, thymus and kidney indexes and antioxidant index verified the success of D-gal aging model. Compared with the premature aging model group, the intervention group Ⅰ had no significant difference in respiratory metabolism, energy consumption, glucose tolerance, insulin sensitivity and other indicators (all P>0.05). But in the intervention group Ⅰ, the activity ability was significantly improved (P=0.022), the activities of SOD (P=0.026) and GSH-Px (P=0.006) in serum were significantly increased, and the MDA content in serum was significantly decreased (P=0.011). Compared with the aging model group, the O2 consumption (P=0.045), CO2 exhalation (P=0.030), activity ability (P=0.049) and energy consumption (P=0.043) in the intervention group Ⅱ were significantly increased at night. Compared with the aging model group, the impaired glucose tolerance was improved (P=0.030), the insulin sensitivity was increased (P=0.010)in the intervention group Ⅱ, the activity of SOD in serum was significantly increased (P=0.046), and the MDA content in serum and liver tissue was significantly decreased (P=0.000). There was no significant difference in the activity of GSH-Px in serum and liver tissue between the two groups (P>0.05).

    Conclusion

    ·NMN can improve the metabolic level of aging mice to a certain extent, and its mechanism may be related to improving the antioxidant capacity of the body.

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    Clinical research
    Establishment of a nomogram clinical scoring system for the risk of heterotopic ossification in patients undergoing surgery after fracture
    Yekai WANG, Wei CHEN, Yinghui YANG, Jingze WU, Heping WANG, Yanzhen YAO, Zhoujun BAO
    2022, 42 (2):  166-172. 
    doi: 10.3969/j.issn.1674-8115.2022.02.005

    Abstract ( 97 )   HTML ( 54 )   PDF (1837KB) ( 176 )  
    Objective

    ·To establish a nomogram scoring system to predict morbidity of heterotopic ossification (HO) in the patients undergoing surgery after fracture.

    Methods

    ·From August 2018 to October 2020, 124 patients with fractures from department of orthopaedics or brain surgery, including 81 males and 43 females with an average age of (56.48±15.45) years old (ranging from 25?91 years old), were enrolled in the study. Clinical features including gender, age, with brain trauma or not, anesthetic mode, operation duration time, and blood loss during operation were collected. Peripheral blood samples were collected at three time points, i.e. 1 d, 7 d and 15 d after fracture, and the sera levels of bone morphogenetic protein 2 (BMP-2), BMP-4, BMP-5, BMP-7, interleukin 4 (IL-4), IL-6, IL-10, interferon γ (IFN-γ), and transforming growth factor-β (TGF-β) were detected by ELISA. All the patients were divided into HO group and non-HO group according to follow-up outcomes. The variables with statistically significant differences (α=0.15) between the two groups selected from clinical and laboratory indications were included in the univariate Cox regression model. Then the variables selected from univariate Cox regression model (α=0.05) were incorporated into multivariate Cox regression model to screen the independent risk factors of HO and calculate the risk ratios (HR). Finally, the nomogram scoring system was output through rms package in R language.

    Results

    ·The follow-up observation found 13 HO cases after a median time of 71 (38, 292) d. Seven independent risk factors for HO, i.e. brain trauma (HR=2.932, P=0.038), operation duration time (HR=1.005, P=0.007), blood loss during operation (HR=1.004, P=0.022), BMP-2 (15d) (HR=1.009, P=0.044), BMP-4 (15 d) (HR=1.004, P=0.011), TGF-β (15 d) (HR=1.011, P=0.046), and BMP-7 (7 d) (HR=1.004, P=0.008), were selected by comparison between groups, univariate COX regression and multivariate COX regression sequentially. The nomogram to predict morbidity of HO after 90 d, 180 d, and 360 d was generated.

    Conclusion

    ·The seven scoring indexes of nomogram scoring system for predicting the risk of HO in the patients undergoing surgery after fracture are with brain trauma or not, operation duration time, blood loss during operation, levels of BMP-2, BMP-4 and TGF-β in peripheral blood 15 d after fracture and BMP-7 7 d after fracture.

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    Difference in recurrence rates after temporomandibular joint disc repositioning surgery with miniscrew anchor at different developmental stages in adolescents
    Xuehong WANG, Xuzhuo CHEN, Yi MAO, Da SHEN, Shanyong ZHANG
    2022, 42 (2):  173-177. 
    doi: 10.3969/j.issn.1674-8115.2022.02.006

    Abstract ( 118 )   HTML ( 51 )   PDF (1598KB) ( 69 )  
    Objective

    ·Based on magnetic resonance imaging (MRI), adolescents with anterior disc displacement (ADD) of temporomandibular joint treated by temporomandibular joint disc repositioning surgery with miniscrew anchor were studied retrospectively to analyze the difference of postoperative recurrence rate among adolescents at different developmental stages.

    Methods

    ·Adolescent patients (12 years old≤age≤18 years old ) who received disc repositioning from January 2015 to January 2020 in the Department of Oral and Maxillofacial Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine were selected and the patients were divided into 2 groups defined by the age of 15. MRIs during at least 6 months follow-up were used to evaluate changes in the disc position and condylar bone. During follow-up, ADD without reduction was considered relapse. The position, length of disc and height of condylar were analyzed quantitatively by modified three-circle method on MRI images and compared by statistical analysis via Software SPSS 25.0. Difference of recurrence rates between the 2 groups was compared by using chi-square test. T test was performed for the difference of condylar growth height between the 2 groups.

    Results

    ·Totally 90 cases were included, including 64 females and 26 males, with the average age of (14.2±1.2) years and follow-up interval of (16.0±12.0) months. Postoperative MRIs showed that all the discs were repositioned. During follow-up, 96.4% (54/56) of the discs in group A (>15 years old) were still in position, whereas 3.6% (2/56) of the discs had relapsed anteriorly. In group B (≤15 years old), 85.2% (75/88) of the discs were still in position, whereas 14.8% (13/88) of the discs had relapsed anteriorly, which was higher than that in group A (P=0.032). The condylar height in group A increased by (1.33±1.29) mm compared with that in group B [(1.79±1.14) mm)]. The condylar height in group B increased more during follow-ups (P=0.036). The degree of anterior disc displacement in group B was greater than that in group A (P=0.002), but there was no significant difference in preoperative disc length between the 2 groups (P=0.585).

    Conclusion

    ·For adolescent patients who received temporomandibular joint disc repositioning surgery with miniscrew anchor, experiencing rapid growth might be a factor affecting the effect of the operation. The postoperative effect of repositioning surgery with miniscrew anchor may be affected by the rapid growth and development of condyle.

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    Characteristics of cerebral perfusion in patients with limb shaking-transient ischemic attacks with different blood pressure variations
    Aiyong YU, Yingchun ZHAO, Xiaochun PAN, Kegui ZHOU, Yuwu ZHAO, Lei GU
    2022, 42 (2):  178-184. 
    doi: 10.3969/j.issn.1674-8115.2022.02.007

    Abstract ( 99 )   HTML ( 49 )   PDF (3470KB) ( 50 )  
    Objective

    ·To investigate whether blood pressure fluctuations in patients with limb shaking-transient ischemic attacks (LS-TIA) are related to the changes of cerebral perfusion and collateral circulation, and explore the pathogenesis primarily.

    Methods

    ·This study was a retrospective cohort study. The patients with LS-TIA who were hospitalized in the Department of Neurology, Shanghai Songjiang District Central Hospital from January 2018 to December 2019 were selected. By using the standard deviation of systolic blood pressure collected by ambulatory sphygmomanometer as the short-term systolic blood pressure variation (SSBPV) index, the LS-TIA patients were divided into 16 cases in high SSBPV group and 14 cases in normal SSBPV group, according to the level of SSBPV. During the same period, 15 patients who went to hospital because of headache or dizziness served as healthy control (HC) group. The patients' age, gender, smoking history, drinking history, body mass index, history of hypertension, history of coronary heart disease, history of hyperlipidemia and history of cerebral infarction were collected. All the patients were anesthetized with 1% lidocaine, and underwent cerebral angiography with a single curved catheter, including bilateral common carotid arteries, bilateral internal carotid arteries, bilateral subclavian arteries, and bilateral vertebral arteries. The number of intracranial arterial stenosis and the patency of collateral circulation were assessed by the surgeon. By single-photon emission computed tomography (SPECT) examination, the region of interest (ROI) was delineated, and the cerebellum was used as a control. The ratio less than 0.9 was used as the criterion for judging lobe hypoperfusion. SPSS 25.0 software was used for statistical analysis. The measurement data conforming to the normal distribution were compared by independent samples t test between the two groups, and the comparison among the three groups was performed by variance analysis. The rest data were qualitative data, the chi-square test was used for comparison between the groups, and P<0.05 was considered statistically significant.

    Results

    ·Compared with the HC group, the total number of intracranial artery stenosis and the number of internal carotid artery stenosis in the high SSBPV group and the normal SSBPV group increased, and the differences were statistically significant (all P=0.000). The number of lobes with hypoperfusion in the high SSBPV group and the normal SSBPV group were higher than that in the HC group (all P=0.000). The number of cases with severe hypoperfusion in the high SSBPV group was higher than that in the HC group (P=0.000). The total number of open collateral circulation in the high SSBPV group was higher than that in the HC group (P=0.001), and the number of secondary collateral circulation in the high SSBPV group was higher than that in the HC group (P=0.014). Compared with the normal SSBPV group, the high SSBPV group had more lobes with hypoperfusion (P=0.041), more severe hypoperfusion cases (P=0.024), and more collateral circulation openings (P=0.034).

    Conclusion

    ·The increased short-term systolic blood pressure variability results in a wider range of cerebral hypoperfusion and a higher proportion of severe cerebral hypoperfusion in the patients with LS-TIA. This may contribute to the pathogenesis of LS-TIA. As a part of the compensatory mechanism, the proportion of openning collateral circulation in the patients is high.

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    Analysis of the effects of individualized one-day outpatient diet guidance on glycemic control, body weight gain rate and pregnancy outcome in pregnant women with gestational diabetes mellitus
    Qin GU, Yingqian XIA, Yiqing ZHU, Liying MA, Lei QU, Wenguang SUN
    2022, 42 (2):  185-191. 
    doi: 10.3969/j.issn.1674-8115.2022.02.008

    Abstract ( 108 )   HTML ( 58 )   PDF (931KB) ( 62 )  
    Objective

    ·To investigate the effects of individual guidance of the energy contribution ratio of meals and total energy on glycemic control, body weight gain rate and pregnancy outcome in pregnant women with gestational diabetes mellitus (GDM) in one-day outpatient.

    Methods

    ·A total of 490 pregnant women admitted to one-day outpatient for GDM of The International Peace Maternity & Child Health Hospital, Shanghai Jiao Tong University School of Medicine from April 2018 to December 2019 were included. According to the energy contribution ratio of meals, they were divided into group A [207 cases, eggs were consumed at mid-morning snack (9:00), and the energy contribution ratio of breakfast and mid-morning snack was 15% and 10%, respectively] and group B [283 cases, eggs were consumed at breakfast (7:00), and the energy contribution ratio of breakfast and mid-morning snack was 20% and 5%, respectively], and the glycemic control levels of the group A and group B were compared. According to the difference of pre-pregnancy body mass index (BMI) and weight gain during pregnancy, after adjusting the dietary energy supply, pregnant women in the group B were further divided into group a (132 cases, no change in total energy) and group b (151 cases, total energy reduced by 1-2 exchange parts). The glycemic control level, body weight growth rate and pregnancy outcome were compared between the group a and group b.

    Results

    ·At 2 h after breakfast and before lunch, there was no significant difference between the blood glucose levels and standard values of pregnant women in the group A and group B. Compared with the group A, the blood glucose level of pregnant women in the group B before lunch had reached the control standard (< 5.60 mmol/L). After lunch, the blood glucose level of pregnant women in the group a was higher than the standard value (< 6.70 mmol/L) (P=0.009), but there was no significant difference between the group b and the standard value. The body weight gain rate of pregnant women in group b after intervention was lower than that before intervention (P=0.004). The incidence of gestational hypertension (P=0.015), premature delivery (P=0.032) and huge mass baby (P=0.032) in the group b was higher than that in the group a.

    Conclusion

    ·Using 20% and 5% energy contribution ratio of breakfast and mid-morning snack for eating is helpful for GDM women to control the blood glucose levels before lunch. At the same time, appropriately reducing the intake of total energy is conducive to GDM women who are overweight or obese before pregnancy, or who gain too much weight during pregnancy to control the blood glucose levels and body weight growth rate after lunch, but they are still prone to adverse pregnancy outcomes. It is suggested that clinical intervention should be carried out as soon as possible.

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    Analysis of clinicopathologic features and prognosis of eight children with granulosa cell tumor of ovary
    Zhengwen XING, Ying WU, Xueli WANG, Qingyu WANG, Wenting WANG, Zhi LI, Bin ZHANG, Jing JIN
    2022, 42 (2):  192-196. 
    doi: 10.3969/j.issn.1674-8115.2022.02.009

    Abstract ( 127 )   HTML ( 50 )   PDF (3077KB) ( 195 )  
    Objective

    ·To investigate the clinicopathological features and prognosis of ovarian granulosa cell tumor (GCT) in children.

    Methods

    ·The clinicopathologic and follow-up data of 8 patients with ovarian GCT in Shanghai Children's Hospital from June 2008 to June 2018 were collected and retrospectively studied to summarize the clinicopathological features and analyze the prognosis.

    Results

    ·The age of onset in this group ranged from 2 to 12 years with median age of 6.5 years. Two cases were adult granulosa cell tumor (AGCT) and six cases were juvenile granulosa cell tumor (JGCT). The first symptoms were abdominal pain or abdominal mass. Six patients were complicated with symptoms of precocious puberty, including 5 cases of true precocious puberty and 1 case of pseudo precocious puberty. The levels of peripheral blood sex hormones changed to varying degrees. Gross examination showed that the tumor masses ranged from 4 to 22 cm in the greatest dimension (average 12.8 cm). All of the 8 tumors were mixed solid-cystic in appearance. Under light microscope, AGCT showed obvious nuclear sulcus and microfollicular structure, containing eosinophilic substances, namely characteristic Call-Exner bodies. JGCT had follicular structures of different sizes, containing basophilic secretions. The inner layer of the follicular wall was composed of granular cells, which could be surrounded by follicular membrane cells. No Call-Exner bodies were found, and nuclear sulcus were rare. The results of immunohistochemistry showed that α-inhibin and CD99 were positive, and epithelial membrane antigen (EMA) were negative in all the cases. In 5 cases calretinin were positive, and in 5 cases cytokeratin (CK) were positive. Ki-67 proliferation index was 5%?50%. There were 3 estrogen receptor (ER) positive cases and 6 progesterone receptor (PR) positive cases. Forkhead transcription factor 2 (FOXL2) was highly expressed in 2 cases of AGCT and 6 cases of JGCT. SRY-box transcription factor 9 (SOX9) positive cells were scattered in 2 cases of AGCT and 3 cases of JGCT. All the cases were treated with standard surgery. Tumor rupture or metastasis was found in 5 cases, and ascites was suspiciously positive in 1 case. Eight children with ovarian GCT were followed up for 19?155 months, and all of them survived in good health.

    Conclusion

    ·The clinical features of AGCT and JGCT in children's ovaries are similar and have typical histopathological features respectively. The selection of suitable immunohistochemical detection method is helpful for differential diagnosis of ovarian tumors in children. The prognosis of ovarian GCT in children is good even with tumor rupture after radical surgery.

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    Evidence-based medicine
    Association between interleukin-1B-511C/T gene polymorphism and coronary atherosclerotic heart disease: a meta-analysis
    Yingchao TAN, Junyue YANG, Lina WANG
    2022, 42 (2):  197-204. 
    doi: 10.3969/j.issn.1674-8115.2022.02.010

    Abstract ( 123 )   HTML ( 52 )   PDF (1841KB) ( 47 )  
    Objective

    ·To evaluate the association between interleukin-1B (IL-1B)-511C/T gene polymorphism and the risk of coronary atherosclerotic heart disease (CHD).

    Methods

    ·Published case-control studies on IL-1B-511C/T gene polymorphism and CHD in PubMed, Web of Science, Scopus and Embase databases were retrieved from the establishment of each database to June 2021. Two researchers independently conducted literature screening and data extraction. The extracted data included the name of the first author, the year of publication, the race of the research objects, matching factors, the source of the control group, genotyping method, sample size, genotype frequency, allele frequency, and whether to meet Hardy-Weinberg equilibrium. Meta-analysis was performed by using Stata 16.0 software.

    Results

    ·A total of 9 literatures were included, including 2 190 patients with CHD and 2 385 controls. Meta-analysis showed that there was no significant correlation between IL-1B -511C/T gene polymorphism and the risk of CHD (T vs C: OR =1.21, 95% CI 0.91?1.61; TT+CT vs CC: OR=1.24, 95% CI 0.88?1.75; CC+CT vs TT: OR=1.28, 95% CI 0.86?1.90). Subgroup analysis showed that compared with individuals with C allele and CC genotype in the Chinese population, the individuals with T allele and TT+CT genotype had a 85% and 116% increased risk of CHD, respectively (T vs C: OR=1.85, 95% CI 1.02?3.36; TT+CT vs CC: OR=2.16, 95% CI 1.10?4.24). In hospital-based case-control studies, compared with individuals with CC+CT genotype, the individuals with TT genotype had a 59% increased risk of CHD (OR=1.59, 95% CI 1.03?2.47).

    Conclusion

    ·There is no significant association between IL-1B-511C/T gene polymorphism and the risk of CHD. In the Chinese population, individuals with T allele and TT+TC genotype have a higher risk of CHD.

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    Techniques and methods
    Effect of DCNN model-assisted colorectal polyp detection system on the detection of colorectal polyps by junior physicians
    Xiaofeng WANG, Lu ZHOU, Leyu YAO, Fan HE, Haixia PENG, Daming YANG, Xiaolin HUANG
    2022, 42 (2):  205-210. 
    doi: 10.3969/j.issn.1674-8115.2022.02.011

    Abstract ( 129 )   HTML ( 56 )   PDF (1476KB) ( 52 )  
    Objective

    ·To explore the effect of a computer-aided detection (CADe) system constructed by a deep convolutional neural network (DCNN) model in artificial intelligence (AI) technology on the detection rate of colorectal polyps among the junior physicians lacking of colonoscopy operation experience.

    Methods

    ·The colonoscopy images from January 2019 to December 2020 and colonoscopy videos from January 2021 to March 2021 from the Endoscopy Center Database of Digestive Endoscopy Center, Tongren Hospital, Shanghai Jiao Tong University School of Medicine were collected. The collected images and videos were divided into dataset 1 (5 908 images) and dataset 6 (360 short videos). Dataset 1 was divided into dataset 1a (4 906 images), dataset 1b (300 images) and dataset 1c (702 images), and dataset 1c was the intercepted images from video dataset 6. Datasets 2 to 5 were the public datasets CVC-ClinicDB, CVC-ColonDB, ETIS-Larib Polyp DB and KVASIR, containing a total of 2 188 images. Dataset 1a and datasets 2?5 were the model training sets, and dataset 1b and dataset 1c were the model testing sets. Ten trained junior physicians with no experience in colonoscopy were randomized into AI-assisted group (Group A, n=5) and non-AI-assisted group (Group B, n=5) to interpret 360 colonoscopy videos and determine the presence of polyps in the videos, respectively. For the first 180 videos, both groups had no AI assistance. For the second 180 videos, the AI-assisted group was supplemented with a DCNN model-assisted colorectal polyp detection system, which processed the video data set through the detection system to mark polyps for the investigator's interpretation, while the non-AI-assisted group watched the original video to interpret the presence of polyps. All the videos were first confirmed by 2 senior endoscopists for the presence of polyps. If there was a dispute, the video was excluded and the diagnosis confirmed by these 2 physicians together was the gold standard. If a polyp was present in the video and the investigator failed to detect it, the diagnosis was considered missed; if no polyp was present in the video and the investigator judged that a polyp was present, the diagnosis was considered misdiagnosed.

    Results

    ·In the first 180 videos without AI assistance, there was no significant difference in the number of missed colon polyps between group A and group B. In the second 180 videos, the number of missed polyps in group A with AI assistance was significantly less than that in group B without AI assistance (P=0.031). Meanwhile, the number of missed polyps in the second 180 videos in group A was less than that in the first 180 videos, and the difference was also statistically significant (P=0.007). In addition, there was no significant difference in the number of misdiagnosed polyps detected between and within the groups.

    Conclusion

    ·The DCNN model-assisted colorectal polyp detection system can significantly improve the polyps detection rate by physicians lacking of colonoscopy operation experience without increasing misdiagnosis of colorectal polyps.

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    Review
    Progress in research on the mechanism of drug resistance to conventional chemotherapeutic drugs in children with acute lymphoblastic leukemia
    Yanyan LIN, Yan XU, Hui LI
    2022, 42 (2):  211-217. 
    doi: 10.3969/j.issn.1674-8115.2022.02.012

    Abstract ( 184 )   HTML ( 221 )   PDF (850KB) ( 103 )  

    Acute lymphoblastic leukemia (ALL) has the highest incidence of hematological tumors in children, accounting for about one-third of all childhood cancer cases. With the continuous optimization of the treatment plan and the expansion of health care coverage in China, remarkable improvements have been achieved in the treatment of childhood ALL, and the overall survival rate of more than 5 years has reached 90%. Lack of effective treatment and unacceptably high recurrence rate are the leading causes of severe decrease in the survival and quality of life of children with drug resistance. In recent years, pathogenesis and regulation mechanism of ALL drug resistance has become a hot topic and a difficulty of research at home and abroad. Studies have shown that children with ALL may not only antagonize a certain chemotherapy (chemotherapeutic) drug, but also have a thorny situation of multi-drug resistance. Therefore, it is of great significance to improve the research on the mechanism of drug resistance of ALL to improve the prognosis of children with ALL. This review summarizes recent research progress in drug resistance mechanisms of conventional chemotherapy drugs for childhood ALL, including hormones, antimetabolite drugs (folate antagonists, thiopurines, and amino acid metabolizing drugs), alkaloids and anthracyclines, in order to provide theoretical basis for coping with clinical drug resistance.

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    Research progress in roles of follicular helper T cells in autoimmune diseases
    Xindi WEI, Xiaoyin NIU
    2022, 42 (2):  218-224. 
    doi: 10.3969/j.issn.1674-8115.2022.02.013

    Abstract ( 177 )   HTML ( 65 )   PDF (1029KB) ( 67 )  

    Follicular helper T cells (Tfh cells) have been defined as a new subset of CD4+ T cells in recent years, mainly expressing surface molecules such as C-X-C motif chemokine receptor type 5, inducible co-stimulator, and secreting cytokine interleukin-21, a key transcription factor of which is B cell lymphoma 6. They exist in the germinal center (GC) of lymphoid tissue and in the peripheral blood as well. With the ability to promote B cell maturation and differentiation, GC formation and antibody production, Tfh cells play important roles in the pathogenesis of many autoimmune diseases, such as systemic lupus erythematosus, rheumatoid arthritis, and primary Sj?gren's syndrome. The abnormal quantity and/or quality of Tfh cells will cause pathological processes like tissue injury and promote disease progression. This article reviews the biological characteristics of Tfh cells and their roles in different autoimmune diseases, which may help to further reveal the pathogenesis of certain autoimmune diseases and provide a new way to treat these diseases by targeting these cells.

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    Effects of food-borne titanium dioxide nanomaterials on digestive tract tissues and gastrointestinal microbiome: a review of recent studies
    Hao WANG, Ran WANG, Qian BA
    2022, 42 (2):  225-229. 
    doi: 10.3969/j.issn.1674-8115.2022.02.014

    Abstract ( 112 )   HTML ( 53 )   PDF (820KB) ( 49 )  

    With the improvement of quality of life and changes of dietary habits, people's requirements for food should not only include food and clothing, but also color, aroma, taste, shape and meaning, in which food additives play a very important role. Titanium dioxide (TiO2) is often widely used as a pigment additive in food and daily necessities, so people will inevitably contact and ingest TiO2 nanomaterials in daily life. Based on this, the article reviews the effect of TiO2 nanomaterials on digestive tract tissues and gastrointestinal microbiome, as well as its direct and indirect pro-inflammatory effects in the occurrence and development of gastrointestinal inflammation, further providing reference improving the safety of TiO2 nanomaterials in the food field.

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    Review of clinical features and treatment of congenital cavitary optic disc anomalies
    Yihua ZOU, Jie PENG, Peiquan ZHAO
    2022, 42 (2):  230-234. 
    doi: 10.3969/j.issn.1674-8115.2022.02.015

    Abstract ( 127 )   HTML ( 153 )   PDF (797KB) ( 51 )  

    Congenital cavitary optic disc anomalies (CCODA) are a group of congenital developmental abnormalities of the eye, which can cause visual impairment. CCODA mainly include morning glory syndrome (MGS), optic nerve coloboma (ONC), optic disc pit (ODP), and peripapillary staphyloma (PS). Clinical features of these abnormalities are similar, and thus accurate differential diagnosis is easily neglected. CCODA can be associated with ametropia, maculopathy and retinal detachment, leading to severer damage to visual function. This paper reviews the latest progresses in the demographic features, pathogenesis, ocular features, systemic abnormalities, differential diagnosis and treatment of CCODA.

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    Research progress in the relationship between abnormal blood glucose fluctuation and cognitive dysfunction of patients with diabetes mellitus
    Rong ZHANG, Li LU, Yaxin WANG, Wenqian DONG, Yu ZHANG, Jian ZHOU
    2022, 42 (2):  235-240. 
    doi: 10.3969/j.issn.1674-8115.2022.02.016

    Abstract ( 118 )   HTML ( 57 )   PDF (867KB) ( 50 )  

    Diabetes can not only cause pathological changes in both large vessels and microvessels, but also accelerate the occurrence and development of cognitive dysfunction, increasing the risk of dementia. Studies have shown that diabetes-induced cognitive dysfunction is associated with elevated blood glucose and abnormal glucose fluctuation, and its pathogenesis is closely related to insulin resistance, oxidative stress, cerebrovascular lesions, inflammatory damage, brain cell apoptosis, as well as amyloid β-protein accumulation. At present, drug therapy for diabetic patients with cognitive dysfunction is still in the exploratory stage. It has been found that some hypoglycemic drugs, such as glucagon like peptide -1 receptor agonist, can improve cognitive dysfunction in these patients. However, it still needs to be further clarified by randomized controlled trails with large sample size. This article reviews the assessment methods of cognitive dysfunction, the effects of abnormal glucose fluctuation on cognitive dysfunction, and the pathophysiological mechanisms involved, as well as the effects of different hypoglycemic agents on cognitive dysfunction in patients with diabetes.

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    Research progress and development trend of lower extremity exoskeleton rehabilitation robot
    Jiyu HAN, Yanhong WANG, Daqian WAN
    2022, 42 (2):  241-246. 
    doi: 10.3969/j.issn.1674-8115.2022.02.017

    Abstract ( 329 )   HTML ( 410 )   PDF (917KB) ( 150 )  

    Lower limb motor dysfunction caused by various causes is an important public health problem in the world today. Lower extremity exoskeleton rehabilitation robot is a new type of wearable bionic device, which is mainly used to realize the standing and walking of patients with lower extremity motor dysfunction. It is a hot research topic in rehabilitation medicine at present. By reviewing the history of lower extremity exoskeleton rehabilitation robot, some breakthroughs and developments are found to have been made in this field in recent years. In the future, if we can overcome the technical problems such as portability, intelligence and modularization, it will be possible to maximize the recovery of patients with lower limb dysfunction. In this paper, the key technologies and clinical applications of wearable lower extremity exoskeleton rehabilitation robot are reviewed comprehensively, and new prospects for the research and development in this field are proposed.

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    Research progress in the function of desmoglein-2 in digestive system tumors
    Yanqing LI, Xiaoxia WANG, Xiangdong JIA, Meng REN, Tianxiang XU
    2022, 42 (2):  247-252. 
    doi: 10.3969/j.issn.1674-8115.2022.02.018

    Abstract ( 164 )   HTML ( 282 )   PDF (823KB) ( 88 )  

    Desmoglein-2 (DSG2) is a member of the cadherin family. As an important part of desmoglein, DSG2 mainly plays a role in maintaining intercellular adhesion. Studies have found that DSG2 can also regulate cell proliferation, differentiation and apoptosis through signal transduction, and plays a key role in regulating intestinal epithelial barrier. With the deepening of research, more and more evidence shows that DSG2 is abnormally expressed in a variety of digestive system tumors, and the expression levels are different. The expression level is closely related to the prognosis of tumors. It is speculated that DSG2 plays different roles in the occurrence and development of different digestive system tumors. This article reviews the biological function of DSG2 and its role in digestive system tumors.

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    Case report
    Clinical manifestation and gene diagnosis of Gitelman syndrome with normal blood magnesium in 1 case
    Jianhua ZHOU, Xiaohua LI, Hongli ZHANG
    2022, 42 (2):  253-258. 
    doi: 10.3969/j.issn.1674-8115.2022.02.019

    Abstract ( 115 )   HTML ( 1147957 )   PDF (2245KB) ( 63 )  
    Objective

    ·To analyze the clinical characteristics and SLC12A3 gene mutation of a patient with normal blood magnesium Gitelman syndrome.

    Methods

    ·A patient with Gitelman syndrome was studied. The clinical manifestations and auxiliary examination results were analyzed, and SLC12A3 gene and CLCNKB gene were detected to make a definite diagnosis.

    Results

    ·The patient developed into an adult with clinical manifestations of low blood potassium, normal blood magnesium and low urinary calcium. The possibility of long-term use of thiazide diuretics, laxatives, chronic diarrhea and vomiting was ruled out, and Gitelman syndrome was diagnosed. The pathogenic mutations c.1084G>A and c.1850G>A in SLC12A3 gene were detected by Sanger direct sequencing, which were confirmed to be the new mutation sites found in this study.

    Conclusion

    ·Gene detection is helpful to early diagnosis of Gitelman syndrome and improves its prognosis.

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