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    Study on the expression of B-cell leukemia/lymphoma 11B in the organ of Corti during mice cochlea development
    JIANG Zhenglin, GAO Yunge, WU Hao
    2022, 42 (10):  1361-1374. 
    doi: 10.3969/j.issn.1674-8115.2022.10.001

    Abstract ( 19 )   HTML ( 3 )   PDF (21161KB) ( 10 )  

    Objective ·To explore the expression of B-cell leukemia/lymphoma 11B (BCL11B) in the organ of Corti during mice cochlea development. Methods ·A total of 24 wild-type C57BL/6J mice of random sex at different ages before and after birth [embryonic 14.5 days (E14.5), E15.5, E16.5 and E18.5, and postnatal 0 day (P0), P2, P4 and P7] were selected, with 3 at each age point. After the mice were killed, the bilateral cochleae of the mice were sampled and fixed. The left cochleae were dissected to obtain complete basement membranes, and right cochleae were dehydrated by sucrose solution following cross section. BCL11B antibody was combined with MYOSIN 6 antibody were used for immunohistochemical analysis of whole mount samples of mouse cochlea basement membrane and cross section samples. Confocal fluorescence microscope was used for observing the tissue samples at different age points after staining, and counting the numbers to calculate the percentage of BCL11B-expressing hair cells. Results ·Whole mount samples of mouse cochlear basement membrane and cross section samples at eight age points were successfully prepared. Confocal fluorescence microscope showed that BCL11B expression was first detected specifically in outer three rolls of MYOSIN6 positive hair cells in E15.5 middle-basal turn of mouse cochlea; with the development of mouse cochlea, BCL11B expression gradually strengthened until P2, at which time point BCL11B reached top expression level and started to weaken in P4, with almost no BCL11B expression detected in P7. During the whole period of BCL11B expression in outer hair cells, no BCL11B expression was detected in other cell types in the organ of Corti. The results of the number and percentage of BCL11B-expressing hair cells showed that there was no significant difference among the three age points of E16.5, P0, and P4 (both P>0.05). Conclusion ·It is confirmed that BCL11B is a specifically expressed protein in the early development of outer hair cells in the organ of Corti by co-immunostaining of BCL11B and MYOSIN6.

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    Basic research
    Effect of intragastric treatment of sodium cromoglycate on dextran sulfate sodium-induced ulcerative colitis in BALB/c mice
    GE Yiqin, HUANG Yuji, LI Weize, LI Yanning, LI Li
    2022, 42 (10):  1375-1382. 
    doi: 10.3969/j.issn.1674-8115.2022.10.002

    Abstract ( 18 )   HTML ( 1 )   PDF (3777KB) ( 10 )  

    Objective ·To explore the role of mast cell membrane stabilizer sodium cromoglycate (Crom) in the modeling of ulcerative colitis (UC) in mice. Methods ·Mice were divided into 3 groups, i.e.control group (n=5), dextran sulfate sodium (DSS) group (n=5) and DSS+Crom group (n=5). Mice in the control group drank water for 7 d, and mice in the DSS group drank 3% DSS for 7 d to establish UC model. Meanwhile, mice in the DSS+Crom group were treated with Crom since the 3rd day during the establishment of UC. The changes of body mass and disease activity index (DAI) scores among the three groups were compared. Hematoxylin-eosin staining (H-E staining) and toluidine blue staining were used to observe the histopathology injury and the number of mast cells in colon tissues. Flow cytometry was used to detect the immune cell infiltration of spleen. Differences among the three groups were compared by using one-way ANOVA. Least significant difference (LSD) was used to further compare the differences between each two groups. Results ·Compared with the control group, the mice in the DSS group had severe hematochezia and intestinal barrier injury, the body weight of the mice in the DSS group decreased significantly (P=0.005), the DAI score increased significantly (P=0.001), the colon length shortened significantly (P=0.003), and toluidine blue staining showed that the number of mast cells in the intestine increased (P=0.000). Moreover, compared with the DSS group, the blood in the stool of the mice in the DSS+Crom group decreased, and the colon length increased (P=0.013); the intestinal pathology showed that the inflammation was improved, and the number of mast cells was reduced (P=0.000). Conclusion ·Stabilizing mast cells with sodium cromoglycate could effectively reduce colon inflammation of mice in UC, which indirectly indicates that mast cells participate in the pathogenesis of UC.

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    Function of UCHL3 in maintaining the survival of FLT3-ITD positive acute myeloid leukemia cells
    HU Jiacheng, ZHU Qian, WANG Jiaqi, WU Yingli, LEI Hu
    2022, 42 (10):  1383-1393. 
    doi: 10.3969/j.issn.1674-8115.2022.10.003

    Abstract ( 10 )   HTML ( 1 )   PDF (4794KB) ( 11 )  

    Objective ·To explore differently expressed deubiquitinases in acute myeloid leukemia (AML) cells with internal tandem duplications of the FLT3 (FLT3-ITD) gene mutation. Methods ·The expressions of some deubiquitinases were detected by Western blotting after transforming exogenous FLT3-ITD or the use of FLT3-ITD inhibitor. The Cancer Genome Atlas (TCGA) public database was used to analyze the expression of UCHL3 in FLT3-ITD-positive patients and its correlation with the prognosis of AML patients. Certain deubiquitinase was knocked down by short hairpin RNA (shRNA) in AML cell lines. Cell counting, CCK-8 assay, flow cytometry and wright stain were used to detect the effect of certain deubiquitinase on the function of AML cells and the combined effect of knockdown of corresponding deubiquitinase or using its inhibitor with all-trans-retinoic acid (ATRA). Results ·Expression of FLT3-ITD in Ba/F3 cells significantly induced the protein expression of UCHL3, while inhibition of FLT3-ITD activity in MOLM-13 and MV4-11 cell lines significantly inhibited the expression of UCHL3 in a time- and concentration-dependent manner. TCGA database analysis showed that there was a significant positive correlation between the expression of FLT3 and the expression of UCHL3 in AML (P=0.000), and a significant negative correlation between the expression of UCHL3 and the survival rate of AML patients (P=0.016). UCHL3 knockdown inhibited proliferation and promoted apoptosis of MOLM-13 and MV4-11 cells, and the expression of differentiation-related transcription factors PU.1 and C/EBPβ were significantly increased. Knockdown of UCHL3 significantly enhanced the apoptosis of MOLM-13 and MV4-11 cells induced by 100 nmol/L ATRA, and the apoptosis-related caspase proteins PARP1, caspase 9 and caspase 3 were significantly cleaved. Meanwhile, UCHL3 knockdown cells showed more features of small nuclear volume, nuclear depression, and rod-shaped or irregular cells after ATRA treatment. The dose-response curves of ATRA and TCID detected by CCK-8 assay showed that UCHL3 inhibitor TCID and ATRA synergistic inhibited FLT3-ITD-positive AML cells survival. Conclusion ·FLT3-ITD up-regulates UCHL3 expression and promotes the survival of AML cells. UCHL3 is negatively correlated with the prognosis of AML patients. Knockdown or inhibition of UCHL3 combines with ATRA to inhibit the survival of FLT3-ITD-positive AML cells.

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    Construction of a prediction model of immune-related long non-coding RNA in gastric cancer
    CHEN Bin, CUI Hongquan, YANG Yijin, XU Haiyan, ZHANG Ling
    2022, 42 (10):  1394-1403. 
    doi: 10.3969/j.issn.1674-8115.2022.10.004

    Abstract ( 18 )   HTML ( 1 )   PDF (7427KB) ( 11 )  

    Objective ·To construct a prediction model of immune-related long non-coding RNA (lncRNA) in gastric cancer patients by bioinformatics method, and explore its application value. Methods ·Transcriptome sequencing (RNA sequencing, RNA-seq) data of 413 gastric cancer samples were downloaded from the cancer genome atlas (TCGA) database, including 32 normal samples and 381 tumor samples. Immune-related genes were obtained from ImmPort website. The immune-related lncRNAs (irlncRNAs) were obtained by correlation analysis. The differentially expressed irlncRNAs (DEirlncRNAs) were obtained by the limma R package, and heat maps and volcano maps were drawn. Batch effects of sample were corrected by constructing DEirlncRNA pairs. The clinicopathological characteristic data of TCGA gastric cancer patients were downloaded, and the DEirlncRNA pairs related to prognosis were obtained by univariate analysis, and screened by LASSO regression analysis. Finally, a risk prediction model was constructed by COX proportional hazards regression analysis. The predictive performance of the model and traditional clinicopathological features were analyzed and compared by calculating the area under the curve (AUC). The patient risk value was calculated according to the formula, and the patients were divided into high and low risk groups according to the optimal cutoff value. Survival maps were drawn by using Kaplan-Meier curves, and differences in survival rates between the two groups were compared by Log-rank test. The relationship between risk scores and clinicopathological characteristics was analyzed according to the Wilcoxon signed-rank test. The independent prognostic factors of gastric cancer patients were verified by univariate analysis and multivariate analysis. The relationship between risk scores and immune infiltrating cells and immune-related genes was validated according to Spearman correlation analysis. The half-maximal inhibitory concentration (IC50) values of drugs in high and low risk groups were compared by using the pRRophetic R package. Results ·Compared with normal tissues, 106 irlncRNAs were differentially expressed in gastric cancer tissues, of which 11 were low-expressed and 95 were high-expressed. A total of 32 DEirlncRNAs pairs were included in the COX proportional hazards model, 20 of which were independent prognostic factors for gastric cancer. The 1-, 2- and 3-year AUC values of the risk prediction model were 0.889, 0.966 and 0.935, respectively, which were significantly higher than those of traditional clinicopathological features. The survival rate of patients in the high risk group was significantly lower than that in the low risk group (P=0.000). High risk scores were more closely related to high tumor stage, distant metastasis and patient death. Univariate analysis showed that age, TNM stage, T stage, N stage, M stage and risk score were closely related to the prognosis of gastric cancer patients (all P<0.05). Multivariate analysis showed that age, TNM stage and risk score were independent prognostic factors for gastric cancer patients (all P<0.05). Risk scores were negatively correlated with various T cells and mast cells, and positively correlated with tumor-associated fibroblasts, macrophages and endothelial cells. In the high risk group, the expression levels of immune-related genes IFNG and MSH2 were lower than those in the low risk group. In the high risk group, the sensitivity of the drugs doxorubicin, cisplatin, tipifarnib and mitomycin were lower than those in the low risk group (all P<0.05). Conclusion ·The COX proportional hazards model constructed with irlncRNAs pairs can accurately predict the survival status and survival rate of gastric cancer patients and its sensitivity to chemotherapy drugs.

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    Effect of chloroquine phosphate on vascular endothelial cell injury induced by double-stranded RNA
    ZHANG Jiaojiao, SUN Junnan, WANG Hairong
    2022, 42 (10):  1404-1412. 
    doi: 10.3969/j.issn.1674-8115.2022.10.005

    Abstract ( 17 )   HTML ( 1 )   PDF (4963KB) ( 11 )  

    Objective ·To investigate the inhibitory effect of chloroquine phosphate (CQ) on vascular endothelial cell injury induced by double-stranded RNA (dsRNA) and its mechanism. Methods ·An in vitro model of vascular endothelial cell injury induced by dsRNA in viral sepsis was established via stimulating human umbilical vein endothelial cells (HUVECs) with polyinosinic-polycytidylic acid (poly I:C). HUVECs were randomly divided into normal control group (NC group), poly I:C group with different concentrations and CQ+poly I:C group. The cell viability of HUVECs before and after poly I:C treatment was detected by using Cell Counting Kit-8 (CCK-8). The morphological changes of HUVECs were observed by using optical microscope. The integrity of cell membrane was assessed by calculating the rate of propidium iodide (PI) positive cells by using flow cytometry. The expression level of mature proinflammatory cytokine interleukin 1β (IL-1β), and key proteins of pyroptosis were detected by Western blotting. Results ·Compared with the NC group, HUVECs treated with poly I:C showed a decreased cell viability, more expanded "Vesicles" and more ruptured cells. An increased level of IL-1β expression and positive rate of PI+ cells were also observed. The expression levels of NOD-like receptor thermal protein domain associated protein 3 (NLRP3), and cleaved caspase-1 and gasdermin D N-terminal (GSDMD-N) were increased significantly (all P<0.05) under the treatment of poly I:C and the changes were concentration-dependent. Interestingly, compared with poly I:C group, the cell viability of HUVEC pretreated by CQ was significantly increased, and a decreased IL-1β level were observed. The phenomena of cell swelling and rupture was improved, and a lower positive rate of PI+ cells was detected. The expression levels of cleaved caspase-1 and GSDMD-N of cell pyrosis were also decreased (all P<0.05). CQ pretreatment did not reduce NLRP3 expression induced by dsRNA, but it did inhibit the expression of Toll-like receptor 3 (TLR3). Conclusion ·Pyroptosis is involved in vascular endothelial cells injury induced by dsRNA, and CQ inhibits dsRNA-induced pyroptosis, which could be related to TLR3 regulation.

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    Clinical research
    Treatment-free remission after imatinib discontinuation in patients with chronic myeloid leukemia
    XU Tianxue, QIAN Ying, LIU Zhanyun, CAI Gang, WU Yingli, LI Junmin, SHEN Zhixiang, ZHOU Li
    2022, 42 (10):  1413-1419. 
    doi: 10.3969/j.issn.1674-8115.2022.10.006

    Abstract ( 17 )   HTML ( 1 )   PDF (1404KB) ( 2 )  

    Objective ·To analyze the outcomes of treatment-free remission (TFR) after imatinib discontinuation in patients with chronic myeloid leukemia in chronic phase (CML-CP) who meet the criteria for TFR and are willing to monitor the disease regularly, the prognostic factors and the application of droplet digital polymerase chain reaction (ddPCR) technique in monitoring TFR. Methods ·The patients with CML-CP enrolled in this study were monitored regularly after imatinib discontinuation. Molecular response and relapse were analyzed by quantitative polymerase chain reaction (QPCR) for detection of BCR-ABL transcripts. The lymphocytes subsets pre- and post-imatinib discontinuation were evaluated by flow cytometry. ddPCR was used to detect BCR-ABL and the predictive role in TFR was analyzed. Results ·① Forty-two CML-CP patients who met the criteria for TFR were assessed. With median follow-up time 41(5?93) months, 32 (76.2%) patients maintained TFR.The estimated TFR rate by 12, 24 and 48 months were 85.1%, 75.1% and 70.1%, respectively. Median TFR duration was 41 (2?93) months. The most common adverse event post-discontinuation was musculoskeletal pain of grade Ⅰ?Ⅱ (31.0%). Eight patients achieved deep molecular response (DMR) after restart of imatinib. ②58.3% of patients with continuous positive ddPCR developed molecular relapse after imatinib discontinuation, while none relapsed in those with negative detection (P<0.01). ③ The percentage of CD8+CD28- cells pre-discontinuation was lower, while the percentage of CD4+CD25+ cells post-discontinuation was higher in relapsed patients than that in TFR patients (6.2% vs 12.6%, P=0.026; 3.2% vs 2.1%, P=0.021). Conclusion ·CML-CP patients who meet the criteria of TFR may successfully maintain TFR after TKI discontinuation. ddPCR may help to predict the outcome of TFR and detect the molecular relapse earlier. Immune regulation by different T cell subsets may play a role in TFR duration to prevent relapse of disease.

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    Weekly paclitaxel in combination with cisplatin in the first-line treatment of metastatic breast cancer: a retrospective study
    WANG Yuzheng, SU Juncheng, TANG Lei, XU Yingchun, ZHANG Fengchun
    2022, 42 (10):  1420-1427. 
    doi: 10.3969/j.issn.1674-8115.2022.10.007

    Abstract ( 15 )   HTML ( 1 )   PDF (1441KB) ( 2 )  

    Objective ·To investigate the efficacy and safety of weekly paclitaxel in combination with cisplatin (DP) as first-line regimen compared with other treatment of physician's choices (TPC) for metastatic breast cancer (MBC). Methods ·The clinical data of 117 MBC patients who were hospitalized in the Department of Oncology, Renji Hospital, Shanghai Jiao Tong University School of Medicine from July 2008 to June 2021 were retrospectively analyzed. Fifty-five patients were divided into DP group and 62 patients were divided into TPC group, according to whether the DP regimen was used as first-line chemotherapy. The DP group received paclitaxel 80 mg/m2 combined with cisplatin 25 mg/m2, administered on the 1st, 8th and 15th day, respectively, every 28 d as a cycle, a total of 6 cycles of treatment. TPC group received other first-linechemotherapy regimens, including single or combined regimens of capecitabine, gemcitabine, docetaxel and other regimens.Clinicopathologic characteristics including age, menopausal status, pathological results and molecular subtypes were collected and analyzed. Objective response rate (ORR) and clinical benefit rate (CBR) were used to evaluate the efficacy of different regimens based on Response Evaluation Criteria in Solid Tumors (RECIST) version 1.1. The adverse effects (AEs) were assessed in accordance with National Cancer Institute's criteria for common adverse effect 5.0 (NCI CTC 5.0), and NCI Ⅲ/Ⅳ AEs were analyzed as severe AEs. The numbers and sites of MBC relapses were analyzed as progression pattern. The differences of clinicopathologic characteristics, efficacy and severe AEs between the two groups were compared by χ2 test. Kaplan-Meier survival analysis was used to compare the difference of first-line progression free survival (PFS1) between the two groups, and the factors with probable influence on PFS1, including clinicopathologic characteristics, treatments and other factors were analyzed. The univariate Cox regression analysis was further applied to analysis of the possible influencing factors screened above. Results ·Except the progesterone receptor (PR) expression (P=0.048), there was no significant difference in clinicopathologic characteristics between the two groups (P>0.05). The ORR (47.3% vs 22.6%, P=0.009) and CBR (78.2% vs 41.9%, P=0.002) in the DP group were significantly higher than those in the TPC group. The median PFS1 was 12.0 months (95%CI 10.0?15.0 months) in the DP group, while the median PFS1 was 6.0 months (95%CI 5.0?9.0 month) in the TPC group, demonstrating significant difference (Log-rank P=0.000). Only the factor "receiving DP regimen as first-line chemotherapy" was recognized as a protective factor for MBC progression by univariate Cox analysis (P=0.000, HR=0.419, 95%CI 0.271?0.649). There was no significant difference (P>0.05) in the progression pattern between the two groups except for fewer bone metastasis (P=0.006). The AEs of the two groups were tolerable, and there was no significant difference in the proportion of severe AEs between the two groups (P>0.05). Conclusion ·The first-line DP regimen demonstrates better efficacy and prolonged PFS1 than TPC regimens. It could be an effective selection and should be considered for MBC according to the clinical situations.

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    Optimum spinal anesthesia dose of ropivacaine combined with a single low-dose S-ketamine during elective cesarean delivery: a double-blind sequential study
    SUN Yuanqing, XU Tao, ZHANG Xiaoyu, XU Zifeng, CHAI Jing
    2022, 42 (10):  1428-1434. 
    doi: 10.3969/j.issn.1674-8115.2022.10.008

    Abstract ( 16 )   HTML ( 1 )   PDF (1467KB) ( 2 )  

    Objective ·To explore the optimal dose of ropivacaine for elective cesarean section under spinal anesthesia combined with a single low-dose of intravenous S-ketamine, and observe the rates of hypotension associated with this method. Methods ·Forty eligible women were included in the study for spinal anesthesia block combined with intravenous low-dose S-ketamine (0.15 mg/kg). The first patient received a 12 mg spinal anesthesia dose of ropivacaine. If the upper sensory block level of the patient was not lower than the sixth thoracic vertebra (T6) and visual analog scale (VAS) score was ≤2 before delivery, the next patient had 1/9 chance of receiving a lower dose or 8/9 chance of receiving the same dose of ropivacaine as the previous patient. If the patient had VAS score>2 or needed an extra epidural analgesics before delivery, a higher dose of ropivacaine was used for the next patient. The primary outcome was whether the effect of spina anesthesia was accurate, that is, the amount of ropivacaine to maintain patients' VAS score ≤2 before delivery, and the incidence of post-spinal anesthesia hypotension. Secondary outcomes mainly included the rate of hypotension-related symptoms and drug interventions, upper sensory block level of anesthesia, level of sedation (Ramsay sedation scale was used as the evaluation standard) and neonatal outcomes. 90% effective dose (ED90) and 95% confidence interval (95% CI) of ropivacaine were estimated by Isotonic regression. Results ·In combination with low-dose S-ketamine, the ED90 of ropivacaine for cesarean section spinal anesthesia was 11.8 mg (95%CI 11.7-12.7). The incidence of hypotension and hypoxemia of forty parturients was 5.0% (2 cases) and 2.5% (1 case), respectively, and there was no bradycardia occurring. All parturients did not use phenylephrine injection as a remedy, and there was no adverse reactions related to hypotension occurring (dyspnea, nausea, vomiting, shiver and blurred vision). There was no reflux and aspiration occurring. Only one parturient before delivery had a Ramsay sedation score of 2 (awake, quiet and cooperative), and the rest scored 3 (sleepy but respond quickly to instruction). At 5 min after delivery, according to the Ramsay sedation scale, 32 parturients were 2 scores, and 8 parturients were 3 scores. At the end of the operation, the Ramsay sedation scale of all parturients was 2 scores. During the cesarean section, the incidence of visceral traction pain, nightmare and dizziness of patients was 7.5%, 62.5% and 52.5%, respectively. In addition, the Apgar scores of 40 newborns at 1 min and 5 min after birth were all 10, and the umbilical vein blood gas analysis showed that the pH was (7.37±0.03). Conclusion ·Combined with a intravenous single low-dose of S-ketamine during spinal anesthesia for elective cesarean section, the ED90 of ropivacaine is 11.8 mg, and this method can effectively reduce the incidence of hypotension.

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    Risk factors of recurrent wheezing in preschool children
    LI Aiqiu, ZHANG Xiaoxiao, JIANG Yunli, XIAO Yanshang, DING Guodong, WU Beirong, DONG Xiaoyan
    2022, 42 (10):  1435-1440. 
    doi: 10.3969/j.issn.1674-8115.2022.10.009

    Abstract ( 11 )   HTML ( 1 )   PDF (1318KB) ( 2 )  

    Objective ·To investigate the potential risk factors of recurrent wheezing in preschool children. Methods ·A case-control study was conducted in 370 children with wheezing admitted to the Department of Respiratory Medicine in a specialized hospital for children in Shanghai from December 2019 to December 2020. The children were preschool children aged 3?6 years. According to the frequency of wheezing, pediatric patients were divided into occasional wheezing group (1?2 attacks per year) and recurrent wheezing group (≥3 attacks per year). The face-to-face questionnaire was performed to the parents responding for basic information and recurrent wheezing risk factors including the children age, gender, gestational age, birth weight, history of eczema, history of allergic rhinitis, history of food allergy, history of inhalational pollen allergy, history of bronchiolitis, maternal age, production mode, feeding pattern, history of parental asthma, resident area (city/village), passive smoking, history of pet contact, seasonal factors, etc. Univariate analysis was used to compare risk factors of recurrent wheezing between the two groups. Logistic regression analysis was used to analyze the independent risk factors associated with recurrent wheezing. Results ·Three hundred and seventy cases were enrolled, including 204 cases (55.14%) in the occasional wheezing group and 166 cases (44.86%) in the recurrent wheezing group. A total of 10 risk factors were determined for recurrent wheezing, which included children's age (χ2=36.608, P=0.000), gender (χ2=4.290, P=0.038), eczema (χ2=6.250, P=0.012), allergic rhinitis (χ2=10.787, P=0.001), inhaling pollen (χ2=8.868, P=0.003), history of bronchiolitis (χ2=27.898, P=0.000), gestational age (χ2=5.141, P=0.023), feeding pattern (χ2=4.316, P=0.038), parental asthma history (χ2=5.050, P=0.025) and resident area (χ2=31.013, P=0.000). The Logistic regression showed that allergic rhinitis (OR=4.759, 95%CI 1.665?13.603), bronchiolitis (OR=12.113, 95%CI 5.686?25.802), premature birth (OR=3.092, 95%CI 1.120?8.539) and living in city (OR=2.395, 95%CI 1.326?4.324) were the independent risk factors of recurrent wheezing. Conclusion ·The independent risk factors for recurrent wheezing in preschoolers include allergic rhinitis, bronchiolitis, premature birth and living in city.

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    Public health
    Chinese version of Fear of Self Questionnaire and its reliability and validity test
    LIU Jinjun, WANG Yuan, WU Yanru, XIAO Zeping
    2022, 42 (10):  1441-1447. 
    doi: 10.3969/j.issn.1674-8115.2022.10.010

    Abstract ( 4 )   HTML ( 1 )   PDF (1698KB) ( 3 )  

    Objective ·To examine the reliability and validity of the Chinese version of Fear of Self Questionnaire (FSQ) among Chinese university students. Methods ·Based on the standard questionnaire translation model, the English version of FSQ was translated into Chinese and the cultural debugging was conducted. A total of 396 students from three science and engineering majors of a comprehensive university in Shanghai from December 2021 to January 2022 were tested with the general information questionnaire, the Chinese version of FSQ, the Chinese version of Ego Identity Scale (EIS), the Chinese version of Self Acceptance Questionnaire (SAQ) and the Chinese version of Obsessive Belief Questionnaire-44 (OBQ-44). The Chinese version of FSQ was tested by item analysis, and the reliability and validity of the Chinese version of FSQ were tested by item analysis, internal consistency reliability, split half reliability, retest reliability, criterion validity, exploratory factor analysis and confirmatory factor analysis. Results ·A total of 386 valid questionnaires were recovered in the initial test, and 163 valid questionnaires were recovered in the retest. A total of 20 items in the Chinese version of FSQ passed the item analysis test, and all items were retained. The internal consistency reliability, split half reliability and retest reliability of the Chinese version of FSQ were 0.931, 0.869 and 0.736, respectively, which met the requirements of metrology. The criterion validity of the Chinese version of FSQ was good, and the total score of the Chinese version of FSQ was significantly correlated with the total score of the three criterion validity questionnaires [Chinese version of EIS (r=-0.279, P=0.000), Chinese version of SAQ (r=-0.568, P=0.000) and Chinese version of OBQ-44 (r=0.418, P=0.000)]. The exploratory factor analysis of the Chinese version of FSQ showed that the questionnaire contained three factors: self fear, self assurance and self questioning. The confirmatory factor analysis of the Chinese version of FSQ showed that the indicators of the three-factor structural equation model fitted well, including χ2/df = 2.790, CFI=0.862, GFI=0.834, AGFI=0.791, RMSEA=0.090, TLI=0.843, NFI=0.802. Conclusion ·The Chinese version of FSQ in this study has good reliability and validity among Chinese university students, and can be used to measure the level of feared self of this group.

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    Investigation and related factor analysis of barriers to outpatient cardiac rehabilitation in patients with coronary heart disease
    LIU Xia, WEN Fule, ZHANG Yaqing
    2022, 42 (10):  1448-1457. 
    doi: 10.3969/j.issn.1674-8115.2022.10.011

    Abstract ( 4 )   HTML ( 1 )   PDF (1489KB) ( 3 )  

    Objective ·To evaluate the current status of barriers to outpatient cardiac rehabilitation (CR) in patients with coronary heart disease (CHD) and analyze related factors. Methods ·From October to December 2017, the inpatients or outpatients with CHD from seven tertiary general hospitals in Yangpu District, Hongkou District, Baoshan District and Songjiang District of Shanghai were selected as the research objects by convenient sampling method. The general information questionnaire, the Cardiac Rehabilitation Information Awareness Questionnaire (CRIAQ) and the Chinese/Mandarin Cardiac Rehabilitation Barriers Scale (CRBS-C/M) were used to investigate the awareness of CR information, the barriers to CR and related factors in the patients with CHD. Results ·A total of 390 questionnaires were distributed, and 342 valid questionnaires were recovered, with an effective recovery rate of 87.7%. The average age of the CHD patients was (67.03±10.83) years old. Among them, 42 patients (12.3%) had heard of CR. The overall median score of CRBS-C/M was 3.10 (2.71, 3.43) points, and the median scores of five dimensions ranged from 3.00 to 3.50 points. The median score of external logistical factors was the highest, which was 3.50 (3.00, 4.00) points. The top three barriers were "distance" "severe weather" and "I did not know about cardiac rehabilitation", with 75.4% (258/342), 69.3% (237/342) and 67.8% (232/342) of the patients agreeing or strongly agreeing, respectively. The average score of CRIAQ was (46.48±12.54) points. Multiple linear regression analysis showed that CRIAQ score (β=-0.242, P=0.000), whether diagnosed as unstable angina or not (β=0.194, P=0.000), gender (β=0.154, P=0.002) and whether participating in outpatient CR programs or not (β=-0.128, P=0.016) were the related factors of CR barriers in the patients with CHD (F=8.909, P=0.000), which explained 25.9% of the total variation. Conclusion ·The CR barrier of the patients with CHD is in the upper middle level, and the biggest barrier factor is the external logistical factor. The top three reasons that patients think hindering their participation in CR are long distance, severe weather and not knowing about CR. The patients with high awareness of CR information, or participating in outpatient CR programs have a low level of CR barriers, while the patients with unstable angina pectoris or the female patients have a high level.

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    Review
    Advances in single-cell RNA sequencing in glomerular diseases
    XING Haifan, FAN Ying
    2022, 42 (10):  1458-1465. 
    doi: 10.3969/j.issn.1674-8115.2022.10.012

    Abstract ( 5 )   HTML ( 1 )   PDF (1670KB) ( 2 )  

    Glomerular diseases are a leading cause of chronic kidney disease and end-stage renal failure. Understanding the molecular mechanism underlying the initiation and progression of glomerular diseases is important to early identify risk factors for disease development and rescue renal function. The kidney is composed of complex and highly differentiated cells, which is a major challenge in the study of glomerular diseases. Bulk RNA sequencing cannot assess differential transcription or the interactions between cell types. Single-cell RNA sequencing (scRNA-seq) technology has developed rapidly in recent years, making it possible to study a wide range of cell types in kidney, urine and blood. The application of scRNA-seq to glomerular diseases helps to construct a comprehensive cell atlas, elucidate complex cellular and molecular mechanisms, develop efficient and accurate prognostic biomarkers and cell-specific therapies. This review focuses on the latest research advances in scRNA-seq in primary and secondary glomerular diseases.

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    Potential role of SIRT1 in unexplained recurrent spontaneous abortion
    LIU Ziwei, CAO Wenwen, WANG Yunrui, FENG Xiaoling
    2022, 42 (10):  1466-1473. 
    doi: 10.3969/j.issn.1674-8115.2022.10.013

    Abstract ( 3 )   HTML ( 1 )   PDF (1311KB) ( 2 )  

    Unexplained recurrent spontaneous abortion (URSA) is also known as alloimmune recurrent abortion. It is suggested that the main pathogenesis of URSA is immune imbalance at the maternal-fetal interface, abnormal trophoblast invasion and abnormal placental angiogenesis. Existing studies have revealed the important role of SIRT1 gene in the field of reproduction and the regulation of immune diseases. However, there is still a lack of systematic and comprehensive research overview on the mechanism of SIRT1 in improving URSA. SIRT1 may affect oxidative stress and autophagy by regulating the acetylation process of histones and key transcription factors, thus participating in numerous feedback loops and networks of URSA occurrence and development. Finally, SIRT1 can play a role in regulating trophoblast cell invasion and maternal-fetal interface angiogenesis. At the same time, the production of proinflammatory cytokines is suppressed to improve the excessive maternal immune inflammatory response during pregnancy. The activity of SIRT1 determines its deacetylation ability, which is crucial for downstream pathways and protein stability. Therefore, improving its activity is of great significance for improving URSA. Studies have shown that metformin and resveratrol can activate SIRT1 and protect SIRT1 and its downstream target proteins, which may be potential therapeutic agents for URSA. Based on the function of SIRT1, this article reviews the potential role of SIRT1 in the pathogenesis of URSA and the exogenous drugs targeting SIRT1 activation, in order to provide reference for the prevention and treatment of URSA in clinical research.

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    Advances in the role of muscle blind-like protein 1 in malignant tumors
    ZHU Nan, LIU Bingya, YU Beiqin
    2022, 42 (10):  1474-1481. 
    doi: 10.3969/j.issn.1674-8115.2022.10.014

    Abstract ( 3 )   HTML ( 1 )   PDF (1339KB) ( 2 )  

    Muscle blind-like protein 1 (MBNL1) is a class of RNA binding protein. As an alternative splicing factor of precursor mRNA (pre-mRNA), it plays a fundamental role in the regulation of the development process and the post transcriptional regulation of specific transcripts, which affects multiple steps in the process of RNA maturation and expression, including pre-mRNA splicing, degradation, RNA output, stability maintenance, modification and translation. MBNL1 was first considered as a contributing factor in the pathogenesis of myotonic dystrophy. With going deep into the research, its role in a variety of non-tumor and tumor diseases has become increasingly prominent. Studies have shown that MBNL1 is abnormally expressed in many types of tumors and is closely related to the occurrence, development and metastasis of various malignant tumors, such as gastric cancer, colorectal cancer, breast cancer, prostate cancer, glioblastoma, lung cancer, hematological system tumors, etc. According to different cell types and development environments, the changes of MBNL1 protein levels in normal and tumor cells are diverse. MBNL1 can not only act as a transcriptional activator to promote the occurrence and development of tumors, but also play as a transcriptional repressor to inhibit the growth and metastasis of tumors. The present review summarizes the molecular mechanism, biological characteristics, expression and function of MBNL1 in the occurrence and development of various malignancies to provide several new insights into tumor-targeted therapy and prognosis evaluation.

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    Initiation and regulatory mechanism of C9ORF72 (G4C2)n RAN translation
    FENG Yiyuan, XU Zhongyun, DING Lin, YIN Yafu, WANG Hui, CHENG Weiwei
    2022, 42 (10):  1482-1489. 
    doi: 10.3969/j.issn.1674-8115.2022.10.015

    Abstract ( 4 )   HTML ( 1 )   PDF (1867KB) ( 2 )  

    As one of the main pathogenic mechanisms of the microsatellite repeat expansion diseases, the repeats from the abnormal DNA expansion produce toxic proteins through repeat-associated non-AUG (RAN) translation, which can causes neuronal death. Amyotrophic lateral sclerosis (ALS) is the most common neurodegenerative disease affecting motor neurons, while frontotemporal dementia (FTD) is a less common early-onset dementia compared to Alzheimer′s disease. C9ORF72 (G4C2)n abnormal expansion is the most common cause of ALS/FTD. Three mechanisms have been proposed for abnormal expansion of (G4C2)n in C9ORF72: ① The C9ORF72 loss-of-function results from the transcription suppression of C9ORF72 caused by the abnormal expansion of (G4C2)n in C9ORF72. ② The RNA foci from the abnormal expansion of (G4C2)n in C9ORF72, which bind to multiple RNA binding protein (RBP), lead to the dysfunction of these RBP. ③ The repeats from the abnormal expansion of (G4C2)n in C9ORF72 undergoing RAN translation produce the dipeptide repeat proteins (DPRs), which results in their toxic gain-of-function. Many studies have evidenced that RAN translation plays a pivotal role in disease progression. While a lot of studies focus on the pathologic mechanism of DPRs, the initiation and regulation mechanism of C9ORF72 (G4C2)n RAN translation is unknown and severely hinders the application of RAN translation as the therapeutic target in ALS/FTD. This review summarizes the most updated literatures on initiation and regulation mechanism of C9ORF72 (G4C2)n RAN translation and discusses the feasibility of reducing cellular toxicity and increasing neuron survival by targeting C9ORF72 (G4C2)n RAN translation.

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    Research progress in the role of Akkermansia muciniphila in gut-related diseases
    JIANG Yi, JIANG Ping, ZHANG Mingming, FANG Jingyuan
    2022, 42 (10):  1490-1497. 
    doi: 10.3969/j.issn.1674-8115.2022.10.016

    Abstract ( 3 )   HTML ( 1 )   PDF (2348KB) ( 2 )  

    Akkermansia muciniphila (A. muciniphila) is one of the normal gut symbionts using mucin as the sole source of carbon and nitrogen elements to maintain colonization and growth. Intestinal homeostasis is crucial for maintaining physiological functions. Intestinal dysfunction is closely related to the occurrence and development of metabolic diseases, immune diseases, infectious diseases and tumors. Gut microbiota is a key factor that influences the intestinal health. As a member of the gut microbes, A. muciniphila plays a convincing role in intestinal inflammation, intestinal tumor and other intestinal disorders involving diseases such as liver diseases and metabolic diseases. The mechanism is under exploration and revealed gradually. Hence, A. muciniphila is considered to be a promising candidate of probiotics. The characteristics of A. muciniphila, its distribution in the gut, its relationship with gut-related diseases and the mechanism are reviewed.

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    Status and prospect of prenatal genetic diagnosis of fetal congenital heart disease
    LI Min, WU Yi, CHENG Weiwei
    2022, 42 (10):  1498-1503. 
    doi: 10.3969/j.issn.1674-8115.2022.10.017

    Abstract ( 2 )   HTML ( 1 )   PDF (1310KB) ( 2 )  

    Congenital heart disease (CHD) is diagnosed as the birth defect that causes heart or blood vessel structure abnormality due to genetic or environmental factors, leading to cardiac dysfunction. It is the most common disease among birth defects in China recently, and its incidence is increasing year by year, seriously affecting the health of newborns, especially for some complicated CHD fetuses. The birth survival rate is very low, and surgical interventions are necessary, which bring heavy psychological and economic burden to the families of fetuses, and seriously affect the aristogenesis and aristogenesis in China. The etiology of CHD is complex. Nowadays, it is believed that the occurrence of CHD is closely related to genetic and environmental factors. Some studies showed that genetic factors were the main causes of CHD, and approximately one of ten fetuses with CHD were related to genetic inheritance. Therefore, prenatal diagnosis to investigate the genetic causes of fetal CHD is crucial for evaluating fetal prognosis. In recent years, more and more studies at home and abroad have focused on the genetic etiology of congenital heart disease. Relevant review articles published in China only focused on the progress of certain genetic testing techniques for the detection of chromosomal abnormalities of CHD, and there is still a lack of comprehensive summary of various prenatal genetic testing techniques on the detection of genetic causes of CHD. This article reviews and prospects various genetic testing techniques for CHD.

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    Case report
    Acute suppurative thyroiditis: a case report and literature review
    WANG Juan, CHEN Qimou, LIU Qicong, CHEN Guangmin, JIANG Peng, ZHANG Rui, ZHOU Houdi
    2022, 42 (10):  1504-1510. 
    doi: 10.3969/j.issn.1674-8115.2022.10.018

    Abstract ( 2 )   HTML ( 1 )   PDF (2191KB) ( 2 )  

    Acute suppurative thyroiditis (AST) is a rare and possibly fatal thyroid disease. A case with "sore throat one month accompanied by left neck mass for two weeks" as the main presentation was reported. The clinical presentation was atypical, and was mistaken for subacute thyroiditis. She was treated with glucocorticoid. The diagnosis of AST was confirmed after ultrasound-guided fine needle aspiration, due to poor efficacy. The features of AST patients over the age of fifty years were also analyzed.

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    A case of protein S deficiency complicated with cerebral embolism and peripheral arterial embolism
    LÜ Mingshun, ZHANG Zihong, WANG Mei
    2022, 42 (10):  1511-1516. 
    doi: 10.3969/j.issn.1674-8115.2022.10.019

    Abstract ( 3 )   HTML ( 1 )   PDF (4926KB) ( 2 )  

    The patient, a 49-year-old male, was admitted to Zhoushan Hospital Affiliated to Wenzhou Medical University in January 2022. He complained of walking instability without obvious inducement for more than 1 month. He had a history of thrombophlebitis and no family history of arterial or venous thrombosis. He had two episodes of acute cerebral infarction, and the tests for thrombophilia in Ruijin Hospital, Shanghai Jiao Tong University School of Medicine showed that the total amount of protein S was 26.7%, and he had a heterozygous mutation C.1955A>T, p.Asp652Val in the exon 15 of PROS1 gene. Then he was diagnosed as having hereditary protein S deficiency (PSD), and thereafter he was given long-term oral rivaroxaban tablets for anticoagulation. This time the physical examinations showed no abnormalities except slightly unsteady walking. The patient scored 6 points in the Venous Thromboembolism (VTE) Risk Assessment Scale (Padua model) with high risk of VTE. Holter testing showed sinus rhythms with occasional premature atrial beats and premature ventricular beats. B-ultrasound for the vessels in the extremities showed multiple thrombus formation with local stenosis in the bilateral popliteal arteries and the posterior tibial arteries, and occlusion in the right radial artery. Echocardiography and carotid ultrasound showed no obvious abnormalities. Cranial MRI showed an old cerebral infarction lesion in the left basal ganglia region. CT angiography combined with three-dimensional reconstruction showed severe stenosis and possible occlusion in the M1 segment of the left middle cerebral artery, and stenosis and sclerosis of the siphon segment of the bilateral internal carotid arteries. Finally, the patient was diagnosed as having PSD with cerebral infarction and peripheral arterials obstruction. He was treated with rosuvastatin calcium for stabilizing plaques, rivaroxaban for anticoagulation, metoprolol succinate for controlling ventricular rhythms, and folic acid for improving metabolism. After 10 d of treatment, the patient 's symptoms were improved.

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