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28 January 2023, Volume 43 Issue 1 Previous Issue    Next Issue
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Basic research
Clinical research
Evidence-based medicine
Techniques and methods
Public health
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Basic research

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Role of methyltransferase like 3 regulating pri-miR-21 methylation in renal fibrosis of diabetes nephropathy WU Jiajin, ZHONG Chen, LI Dawei, CHEN Ruoyang, QU Junwen, ZHANG Ming 2023, 43 (1):  1-7.  doi: 10.3969/j.issn.1674-8115.2023.01.001 Abstract ( 458 )   HTML ( 50 )   PDF (2468KB) ( 459 )   Objective ·To investigate the role of methyltransferase like 3 (METTL3) acting on N6-methyladenosine (m6A) and regulating pri-miR-21 methylation in the renal fibrosis of diabetic nephropathy (DN). Methods ·Eight-week-old male db/db mice were used as DN models, and db/m mice were used as controls. The mice were randomly divided into 4 groups according to whether they received the treatment of 3-deazaadenosine (DAA) by tail vein injection or not (5 mice/group): db/m group, db/db group, db/m+DAA group and db/db+DAA group. From the age of 8 weeks, DAA was injected once per 5 d for a total of 8 times. After the DAA intervention, the mice were kept until they were 19 weeks old. The blood, the urine and the kidney tissue samples of the mice were collected, and blood glucose (BG), serum creatinine (Scr), and urinary albumin-to-creatinine ratio (ACR) were detected. The kidneys were stained with hematoxylin-eosin (H-E), Masson and sirius red to observe the pathological changes. The methylation level of m6A in total RNAs of the kidney was detected with the kit. The expression levels of METTL3 and fibrosis-related proteins in the kidney were detected by Western blotting. The overall pri-miR-21 and the mature miR-21 were detected by real-time quantitative PCR. After enrichment of the m6A-methylated RNAs in the kidney by immunomagnetic beads, the methylated pri-miR-21 at m6A was detected by PCR. Results ·Compared with the db/m group, the levels of BG, Scr, and ACR, and METTL3, m6A methylation level, fibrosis-related proteins, overall pri-miR-21, m6A-methylated pri-miR-21 and mature miR-21 in the kidney in the db/db group significantly increased (P<0.05). Furthermore, the mesangial matrix in the kidney increased, glomerular basement membrane thickened, and the accumulation of collagen fibers increased significantly in the db/db group. Compared with the db/db group, the levels of BG, Scr, and ACR, and m6A methylation level, fibrosis-related proteins, m6A-methylated pri-miR-21 and mature miR-21 in the kidney in the db/db+DAA group decreased significantly (P<0.05) and the degree of renal injury and fibrosis was significantly reduced, but the expression level of overall pri-miR-21 significantly increased (P=0.000). The expression level of METTL3 protein did not change significantly. Conclusion ·The m6A methylation modification of pri-miR-21 promotes the maturation of miR-21, thereby promoting the occurrence and development of renal fibrosis in DN mice; inhibition of METTL3 can inhibit renal fibrosis in DN mice by regulating m6A methylation of pri-miR-21. Figures and Tables | References | Related Articles | Metrics

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Preventive effect of icariin on transition from acute kidney injury to chronic kidney disease in mouse model XIE Lin, CHENG Ye, ZHENG Qimin, ZHANG Xi, FU Lili, CHEN Min, WANG Yi, MEI Changlin, XIE Jingyuan, GU Xiangchen 2023, 43 (1):  8-19.  doi: 10.3969/j.issn.1674-8115.2023.01.002 Abstract ( 459 )   HTML ( 42 )   PDF (7150KB) ( 461 )   Objective ·To investigate the effect of icariin on the transition from acute kidney disease (AKI) to chronic kidney disease (CKD) in mice and the possible mechanism. Methods ·C57BL/6 mice were intraperitoneally injected with aristolochic acid for 2 weeks to mimic AKI to CKD transition. Icariin was intragastrically administered for 2 or 5 weeks at the end of 2 weeks of aristolochic acid injection or 1 week before modeling. The renal function and pathological injury of the mice were monitored. The mitochondrial structure of renal tubular epithelial cells was observed by transmission electron microscopy. The indexes related to fatty acid oxidative metabolic pathway, fibrosis, and inflammation were detected by quantitative PCR (qPCR) and immunohistochemical staining. The rat renal fibroblast cell line and the rat renal tubular epithelial cell lines were co-cultured in Transwell cell chambers to simulate the interaction between the two types of cells. The renal tubular epithelial cells were pretreated with icariin for 1 h, and then treated with aristolochic acid for 24 h. After removing the two drugs, the renal tubular epithelial cells and the fibroblasts were co-cultured for 24 h. The mRNA or protein expression levels of peroxisome proliferator-activated receptor α (Ppara), pro-fibrotic factors, inflammatory factors, activated caspase-3 (cleaved caspase-3), and extracellular matrix-associated genes in renal fibroblasts were detected by qPCR or Western blotting. The renal tubular epithelial cells were transfected with Ppara-siRNA to observe the expression changes of cleaved caspase-3 and the related indicators of mitochondrial fatty acid oxidative metabolism in the downstream of PPARα after icariin intervention prior to aristolochic acid. Results ·Compared with the control group, the serum creatinine and urea nitrogen levels of the mice injected with aristolochic acid only were significantly higher, with a large number of inflammatory cells infiltrating into the kidney and serious renal tubular injury at the end of 2 weeks of aristolochic acid injection and 2 weeks after the last injection (4 weeks). The renal function was significantly improved, and the pathological damage was attenuated in the mice pretreated with icariin. After 4 weeks of modeling, the mitochondrial structure of renal tubular epithelial cells was seriously damaged in the aristolochic acid group, and the mRNA levels of fatty acid oxidative metabolic pathway-related genes were significantly downregulated, compared to the control group, accompanied by macrophage infiltration and renal fibrosis. Icariin pretreatment attenuated these damages generated by aristolochic acid. The serum creatinine and urea nitrogen levels, renal pathological injury, and fibrotic markers of the mice treated with icariin at the end of 2 weeks of aristolochic acid injection showed no difference from the aristolochic acid group. In the in vitro experiments, aristolochic acid treatment suppressed Ppara mRNA expression and elevated cleaved caspase-3, inflammatory factors, and pro-fibrotic factors in renal tubular epithelial cells. It also upregulated the expression of extracellular matrix-related genes in co-cultured renal fibroblasts. In contrast, icariin pretreatment abrogated the changes caused by aristolochic acid. The knockdown of Ppara by siRNA in renal tubular epithelial cells removed the preventive effects of icariin on aristolochic acid inhibiting fatty acid oxidative metabolism and promoting cell apoptosis. Conclusion ·Icariin prevents AKI to CKD transition induced by aristolochic acid in mice. It may play its protective role by improving the mitochondrial fatty acid oxidative metabolic pathway, especially through PPARα, in renal tubular epithelial cells. Figures and Tables | References | Related Articles | Metrics

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MicroRNA-30b-5p inhibits autophagy in ovarian granulosa cells in polycystic ovary syndrome rats by targeting Atg5 WANG Xuemin, WANG Yanan, NIU Aiqin, YE Ying, LI Fei 2023, 43 (1):  20-28.  doi: 10.3969/j.issn.1674-8115.2023.01.003 Abstract ( 314 )   HTML ( 28 )   PDF (4486KB) ( 272 )   Objective ·To explore the expression of microRNA-30b-5p (miR-30b-5p) in polycystic ovary syndrome (PCOS) rats and the effect of miR-30b-5p overexpression on ovarian granulosa cell (GC) autophagy. Methods ·Dehydroepiandrosterone (DHEA) was performed to establish a PCOS rat model, and real-time fluorescent quantitative PCR (qRT-PCR) and Western blotting were performed to measure the expression of miR-30b-5p and autophagy-associated protein 5 homologue (Atg5) in the ovarian tissues of the normal group and PCOS group. Primary PCOS rat ovarian GCs were isolated and cultured, and divided into control group, miR-NC group, miR-30b-5p overexpression group, miR-30b-5p overexpression+pcDNA3.1-NC group, and miR-30b-5p overexpression+pcDNA3.1-Atg5 group. In addition, the ovarian GC of the normal group was taken as the blank group. MiR-30b-5p mimic, pcDNA3.1-Atg5 and the corresponding negative control were transfected into the cells, and 48 h after transfection, qRT-PCR was performed to measure the expression of miR-30b-5p and Atg5 mRNA of cells in each group to verify the transfection effect. CCK-8 and flow cytometer were performed to measure cell viability and apoptosis rate, respectively; immunofluorescence staining was performed to measure the positive expression of microtubule-associated protein 1 light chain 3 (LC3) in each group. Western blotting was performed to measure the protein expression of autophagy-related proteins Atg5, p62, Beclin-1 and LC3. Results ·The expression level ofmiR-30b-5pin the ovarian tissue of the PCOS group was significantly lower than that of the normal group, and the levels of Atg5 mRNA and protein were significantly higher than those of the normal group (all P=0.000). After transfection, compared with the blank group, the miR-30b-5p level, apoptosis rate, and p62 protein level in the ovarian GC of the control group were significantly reduced, the Atg5 mRNA and protein levels, cell proliferation activity, LC3 positive cell percentage, Beclin-1 protein level and LC3Ⅱ/LC3Ⅰ ratio were significantly increased (all P=0.000). Compared with the control group, the miR-30b-5p level, apoptosis rate, and p62 protein level in the ovarian GC of the miR-30b-5p overexpression group were significantly increased, and the Atg5 mRNA and protein levels, cell proliferation activity, LC3 positive cell percentage, Beclin-1 protein level and LC3Ⅱ/LC3Ⅰ ratio were significantly reduced (all P=0.000). Up-regulation of Atg5 can significantly attenuate the inhibitory effect of miR-30b-5p overexpression on ovarian GC proliferation and autophagy (all P=0.000). Conclusion ·MiR-30b-5p is lowly expressed in PCOS; overexpression of miR-30b-5p can inhibit the proliferation and autophagy of ovarian GC in PCOS rats, and promote cell apoptosis, and its mechanism may be related to the inhibition of Atg5 expression. Figures and Tables | References | Related Articles | Metrics

Clinical research

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Characteristics and clinical significance of serum renalase in patients with acute ischemic stroke JIANG Wenqun, HOU Pinpin, CHEN Yan, JIA Feng, ZHANG Xiaohua, GAO Li, HU Qin 2023, 43 (1):  29-35.  doi: 10.3969/j.issn.1674-8115.2023.01.004 Abstract ( 384 )   HTML ( 24 )   PDF (1534KB) ( 304 )   Objective ·To examine the level change of serum renalase in the patients with acute ischemic stroke (AIS), and analyze its role in evaluating disease. Methods ·A total of 118 AIS patients admitted to the Department of Neurology, Renji Hospital, Shanghai Jiao Tong University School of Medicine from July 2020 to November 2021 were enrolled in the case group (AIS group). The patients were assessed for neurological deficits according to the National Institutes of Health Stroke Scale (NIHSS), and were classified as mild and moderate-severe neurological deficits. Another 133 healthy people who participated in physical examination in the physical examination center of the hospital during the same period were selected as the control group. The serum renalase levels of the two groups were detected by ELISA. Spearman's rank correlation analysis was used to evaluate the correlation between the levels of serum renalase and gender, age, fasting blood glucose, blood lipids and NIHSS scores in the patient with AIS. The predictive value of renalase expression level in AIS diagnosis was analyzed by receiver operator characteristic (ROC) curve. The factors that were statistically significant in the results of the univariate Logistic regression analysis were included in the multivariate Logistic regression model. Results ·The level of serum renalase in the AIS group was 2 960.01 (1 557.99, 4 053.70) pg/mL, which was higher than 821.02 (391.29, 1 752.70) pg/mL in the control group, with a statistically significant difference (P=0.000). Spearman's rank correlation analysis showed that the levels of serum renalase in the patients with AIS were negatively correlated with the NIHSS scores (r=-0.216, P=0.019), positively correlated with the serum fasting glucose (r=0.200, P=0.030), and not significantly correlated with gender, age, low-density lipoprotein levels, total cholesterol levels, and the presence of hypertension, diabetes, or coronary heart disease. Serum renalase levels were higher in the AIS with mild neurological deficit patients than those in the moderate-severe deficit patients, and the difference was statistically significant (P=0.034). The ROC curve showed that the cut-off value of serum renalase level to diagnose AIS was 1 856.49 pg/mL, the area under the curve was 0.777±0.030 and its 95%CI was 0.718?0.836 (P=0.000). Multivariate Logistic regression analysis showed that elevated serum renalase level [>1 856.49 pg/mL, odds ratio (OR)=6.980, P=0.000], hypertension (OR=5.382, P=0.000), and diabetes (OR=2.453, P=0.040) were risk factors for AIS. Conclusion ·Serum renalase level is significantly elevated in AIS patients, and negatively correlated with NIHSS score. Serum renalase might be a potential biomarker for the auxiliary diagnosis and assessment of AIS, providing new ideas for the assessment of stroke disease progression and precise treatment. Figures and Tables | References | Related Articles | Metrics

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Effect of short-term GnRH pulse therapy on pituitary-testicular function in adolescent male patients with congenital hypogonadotropic hypogonadism WANG Fei, GONG Yan, XU Liya, LIU Qingxu, LI Yan, GUO Sheng, LI Pin 2023, 43 (1):  36-43.  doi: 10.3969/j.issn.1674-8115.2023.01.005 Abstract ( 375 )   HTML ( 17 )   PDF (1820KB) ( 516 )   Objective ·To investigate the effect of short-term gonadotropin-releasing hormone (GnRH) pulse therapy on pituitary and testicular function in the adolescent male patients with congenital hypogonadotropic hypogonadism (CHH). Methods ·A retrospective study was conducted on 20 adolescent male patients with CHH who received GnRH pulse therapy from January 2016 to June 2021, and their clinical data were collected. They were treated with subcutaneous continuous pulsed administration of gonadorelin by the pump for 1 week (20 cases), of which 5 cases were treated for 3 months. The dose was 8?10 μg per pulse, and the pulse interval was 90 min. The levels of luteinizing hormone (LH), follicle stimulating hormone (FSH) and testosterone were measured before GnRH pulse therapy at 1 week, 1 month and 3 months after treatment. After 3 months of treatment, the testicular volume was measured. All 20 patients with CHH underwent whole exome sequencing. Results ·The age of 20 CHH patients was 14.35 (14.08, 15.31) years old. The clinical manifestations were infantile testis (20/20) and micropenis (20/20), followed by obesity (12/20), dysosmia (9/20), insulin resistance (4/20), cryptorchidism (4/20), and short stature (3/20). The patients' height was 161.79 (154.90, 173.25) cm, body mass index was 23.80 (20.51, 27.46) kg/m2, and testicular volume was 0.91 (0.55, 1.25) mL. Inhibin B was 39.67 (11.29, 64.97) pg/mL; the base values of LH, FSH and testosterone before therapy were 0.20 (0.10, 0.30) IU/L, 0.87 (0.23, 0.89) IU/L, and 0.92 (0.38, 1.49) nmol/L, respectively. After 1 week of continuous GnRH pulse therapy, the base and peak values of LH and FSH and the peak value of testosterone in the 20 patients significantly increased (all P<0.05). In the 5 patients treated for 3 months, the base values and peak values of LH and FSH gradually increased with the prolongation of treatment time. After 3 months of treatment, the base values and peak values of LH and FSH, and the peak value of testosterone were significantly higher than those before treatment (all P<0.05), and the testicular volume was also significantly increased (P=0.004). Gene mutations were detected in only 14 of 20 patients, including fibroblast growth factor receptor 1 (FGFR1) mutations in 7 cases, anosmin 1 (ANOS1) mutations in 4 cases, prokineticin receptor 2 (PROKR2) mutations in 2 cases, and a prokineticin 2 (PROK2) mutation in 1 case. There was no significant difference of the effect of GnRH pulse therapy for 1 week on pituitary-testicular function between the patients with FGFR1 mutations and ANOS1 mutations. Conclusion ·The continuous GnRH pulse therapy for 1 week can make pituitary-testicular function respond in adolescent male CHH patients; the treatment for 3 months helps to induce the secondary sexual characteristics of puberty. Figures and Tables | References | Related Articles | Metrics

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A case-control study of the relationship between early-life environmental exposure and childhood asthma DENG Yuntian, XIONG Wenkui, ZHU Rui, LIU Enmei, LI Xuemei, ZHONG Zhaohui 2023, 43 (1):  44-51.  doi: 10.3969/j.issn.1674-8115.2023.01.006 Abstract ( 419 )   HTML ( 24 )   PDF (1414KB) ( 430 )   Objective ·To explore the potential impact of early-life environmental exposure on childhood asthma in Chongqing, China. Methods ·A case-control study was designed. The cases with asthma diagnosis were enrolled from outpatients of the respiratory medicine departments and the healthy children without history of asthma were enrolled from health check-up clinics of the child health care departments in two tertiary children's hospitals in Chongqing from September 2020 to January 2022. The children in the two groups had all lived in Chongqing since birth and their home addresses had not changed before they were 3 years old. A self-developed “Children's Early-Life Environment Survey” was used to collect general personal data, family information, child health status, birth history, and indoor environment from birth to 3 years old (second-hand smoke, dampness and mold points in bedroom, seen cockroaches in bedroom, bedroom cleaning frequency, air conditioning and air purifier use, and decoration). Based on the home address information before 3 years old, annual particular matter 2.5 (PM2.5) exposure levels were estimated by using a high spatiotemporal resolution model. Univariate and multivariate Logistic regression models were used to analyze the early-life environmental factors affecting the development of childhood asthma. The risk factors which were statistically significant in univariate Logistic regression or had clinical significance were included in the multivariate model. Results ·A total of 220 asthma cases and 636 healthy control children were enrolled. The mean age of the asthma cases and the controls were (7.4±2.1) and (7.6±2.1) years old, respectively. There were no statistically significant differences in age, gender, gestational age, birth weight, mode of delivery, family size, annual family income, maternal education level and living space per person. Multivariate Logistic regression analysis showed that early-life bedroom dampness and mold exposure [odds ratio (OR)=2.155, 95% confidence interval (CI) 1.304?3.559, P=0.003], bedroom cockroach exposure (OR=1.830, 95%CI 1.287?2.601, P=0.001), bedroom air conditioner use (OR=2.328, 95%CI 1.098?4.937, P=0.028), second-hand smoke exposure (OR=1.762, 95%CI 1.272?2.440, P=0.001), and long term exposure to PM2.5 at one year old (OR=1.063, 95%CI 1.034?1.093, P=0.000) increased the risk of childhood asthma. Daily use of air purifier (OR=0.416, 95%CI 0.213?0.812, P=0.010) could reduce the risk of childhood asthma. Conclusion ·Early-life environmental exposure is of great significance for the development of childhood asthma. Early-life bedroom dampness and mold exposure, cockroach exposure, second-hand smoke, incorrect use of air conditioner, and long-term exposure of children to PM2.5 in the first year after birth are independent risk factors for the development of childhood asthma. Figures and Tables | References | Related Articles | Metrics

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Construction and evaluation of a nomogram prediction model for bacterial infection in patients with decompensated hepatitis C cirrhosis XUE Linlin, LI Binghan, CHANG Lixian, LI Weikun, LIU Chunyun, LIU Li 2023, 43 (1):  52-60.  doi: 10.3969/j.issn.1674-8115.2023.01.007 Abstract ( 354 )   HTML ( 25 )   PDF (2418KB) ( 222 )   Objective ·To explore the influencing factors of bacterial infection in decompensated stage of hepatitis C cirrhosis, and establish a risk prediction model of nomogram. Methods ·A total of 574 patients with decompensated hepatitis C cirrhosis were retrospectively collected from The Third People′s Hospital of Kunming between January 2020 and December 2021, and divided into non-infected and infected groups according to whether bacterial infection occurred. The general information, complications, and laboratory indicators were collected. The variables were screened by univariate analysis, and least absolute shrinkage and selection operator (LASSO) regression, and the nomogram model were constructed and verified by multivariate Logistic regression analysis of influencing factors. The decision curve and clinical impact curve (CIC) were used to evaluate the clinical application value of the model. Results ·Bacterial infections occurred in 28.4% (163/574) of the patients, with a total of 191 sites, mainly including spontaneous bacterial peritonitis (86/191) and pulmonary bacterial infections (79/191). Totally 78 strains of pathogens were isolated and cultured, mainly including Klebsiella pneumoniae (15/78) and Escherichia coli (15/78). Multivariate Logistic regression analysis showed that age ≥60 years [odds ratio (OR)=2.054, 95% confidence interval (CI) 1.104?3.822, P=0.023], female (OR=1.701, 95%CI 1.112?2.602, P=0.014), ascites (OR=2.386, 95%CI 1.601?3.557, P=0.000), history of invasive procedures in the last two weeks (OR=2.605, 95%CI 1.368?4.960, P=0.004), and hospitalization time≥2 weeks (OR=1.629, 95%CI 1.098?2.416, P=0.015) were independent risk factors for bacterial infection in decompensated hepatitis C cirrhosis patients, while infusing human serum albumin (OR=0.324, 95%CI 0.194?0.542, P=0.000) and high level of total cholesterol (OR=0.675, 95%CI 0.549?0.830, P=0.000) were protective factors. The nomogram model was constructed with the above seven influencing factors. Receiver operator characteristic (ROC) curve analysis showed that the area under the curve (AUC) was 0.736 and the sensitivity was 80.4%; and the specificity was 65.1%. Hosmer-lemeshow test showed that the model had a good degree of fit (χ2=9.030, P=0.340). The bootstrap method was used for internal repeated sampling for 1 000 times, the average absolute error was 0.010, the calibration curve and the ideal curve were basically fitted, and the predicted values were in good agreement with the actual values. The decision curve showed that the nomogram model had certain clinical practicability in the high risk threshold range (0.040?0.715). CIC showed that the nomogram model can be used to forecast the high-risk population in different levels. Conclusion ·The nomogram risk prediction model constructed in this study has good predictability, consistency and clinical practicability, and can provide evidence for clinicians to preliminary judge the risk of bacterial infection in patients with decompensated hepatitis C cirrhosis. Figures and Tables | References | Related Articles | Metrics

Evidence-based medicine

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A meta-analysis of prevalence of mouth opening restriction in patients with oral cancer YANG Ling, HOU Lili, ZHAO Yan, CHEN Weihong, ZHANG Jinfeng, MAO Yan 2023, 43 (1):  61-69.  doi: 10.3969/j.issn.1674-8115.2023.01.008 Abstract ( 327 )   HTML ( 16 )   PDF (1850KB) ( 532 )   Objective ·To study the prevalence of mouth opening restriction in oral cancer patients by meta-analysis. Methods ·China National Knowledge Infrastructure (CNKI), Chinese Science and Technology Journal Database (VIP), Wanfang Data (Wanfang), Chinese BioMedical Literature Database (CBM), PubMed, Ovid, EMbase, Cochrane Library, Web of Science/SciSearch, and ClinicalTrials.gov were searched for observational studies about the prevalence rate of restricted mouth opening in oral cancer patients (including prospective, retrospective, and cross-sectional studies). The time limit for retrieval was from the database establishment to April 30th, 2022. The obtained documents were independently screened and extracted according to the inclusion and exclusion criteria, and the literature quality evaluation was performed by using methodological index for non-randomized studies (MINORS). The data were combined with the mouth-opening-restricted prevalence rate as the outcome indicator, the meta-analysis was performed by using Stata 15.0 software, and the subgroup analysis was performed based on the detection time, tumor site, and tumor stage. The evidence quality of outcome indicators was evaluated by using GRADE profiler 3.6 software according to GRADE (Grading of Recommendations Assessment, Development, and Evaluation) system. Results ·A total of 3 719 documents were obtained during the initial examination, and 18 documents were finally included, including 2 701 objects. The meta-analysis results showed that the total prevalence rate of mouth opening restriction in oral cancer patients was 42.7% (95%CI 27.0%?59.1%). The results of subgroup analysis showed that the prevalence rates of mouth opening restriction in patients with oral cancer before surgery, and 3 months, 6 months and 1 year or more after surgery were 29.6% (95%CI 9.0%?55.8%), 75.2% (95%CI 68.3%?81.5%), 59.1% (95%CI 38.8%?77.9%) and 22.3% (95%CI 2.0%?54.3%), respectively. The prevalence rates of mouth opening restriction in patients with oral cancer at T1?T2 stage and T3?T4 stage were 38.6% (95%CI 26.0%?52.0%) and 99.6% (95%CI 93.9%?100.0%) respectively. The prevalence rates of mouth opening restriction in patients with oral cancer in the retromolar triangle, the gingiva, the tongue, the salivary gland, the floor of mouth and the lip were 93.1% (95%CI 68.5%?100.0%), 68.1% (95%CI 52.9%?81.7%), 46.1% (95%CI 11.6%?82.7%), 26.1% (95%CI 15.9%?37.5%), 21.9% (95%CI 0.7%?54.0%), and 3.1% (95%CI 0?9.3%), respectively. The GRADE system for evidence quality grading indicated that the quality of evidence for outcome indicator was extremely low. Conclusion ·The total prevalence rate of mouth opening restriction in patients with oral cancer is high, nearly 50%; the patients at 3 months after surgery, at 6 months after surgery, at T3?T4 stage or whose tumor located in the retromolar triangle have higher prevalence rates. Figures and Tables | References | Related Articles | Metrics

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Effect of prehabilitation on outcomes in patients undergoing elective gastrointestinal surgery: a systematic review FANG Fang, TAI Rui, YU Qian, ZHANG Yaqing 2023, 43 (1):  70-78.  doi: 10.3969/j.issn.1674-8115.2023.01.009 Abstract ( 354 )   HTML ( 19 )   PDF (3232KB) ( 274 )   Objective ·To systematically evaluate the effect of prehabilitation on outcomes in patients undergoing elective gastrointestinal surgery. Methods ·PubMed, EMbase, Cochrane Library, Web of Science, CINAHL, Chinese BioMedical Literature Database (CBM), China National Knowledge Infrastructure (CNKI), Wanfang and VIP Database were searched systematically from the establishment of each database to January 31, 2022. The literatures were screened, and evaluated according to the preset inclusion and exclusion criteria. The literature quality was evaluated and the data were extracted. Meta-analysis was performed on randomized controlled trials by using RevMan 5.4. The main observation indexes were the postoperative complication rate and the incidence of surgical site infection. The secondary indexes were hospitalization days, 6-minute walk test (6MWT), Hospital Anxiety and Depression Scale (HADS) score, mortality and compliance of prerehabilitation execution. Results ·Sixteen articles were included, including 15 articles in English and 1 article in Chinese, with a total of 1 616 patients. The overall quality of the included researches was good. The meta-analysis results showed that compared with the control group, the incidences of postoperative complications and surgical site infection were reduced [(odds ratio, OR)=0.57, 95% (confidence interval, CI)0.35?0.94, P=0.030; OR=0.64, 95%CI 0.46?0.90, P=0.009]; the length of hospital stay was shortened [mean difference (MD)=-2.45, 95%CI -3.17 ? -1.73, P=0.000] in the experimental group after the implementation of prehabilitation. There was no significant difference in preoperative 6MWT level between the two groups. Compared with the baseline, the change of 6MWT level before operation of the experimental group was bigger than that of the control group (MD=24.19, 95%CI 3.77?44.60, P=0.020). There were no significant differences in the preoperative HADS score and the postoperative mortality between the two groups. Conclusion ·Prehabilitation can reduce the incidence of postoperative complications, especially surgical site infection, in patients undergoing elective gastrointestinal surgery, thus shortening the length of hospital stay and promoting the recovery of patients. Figures and Tables | References | Related Articles | Metrics

Techniques and methods

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Establishment of a 3D culture model in vitro of pancreatic cancer primary cells using hydrogel microspheres MA Fangfang, QIN Jiejie, REN Lingjie, TANG Xiaomei, LIU Jia, SHI Minmin, JIANG Lingxi 2023, 43 (1):  79-87.  doi: 10.3969/j.issn.1674-8115.2023.01.010 Abstract ( 362 )   HTML ( 21 )   PDF (6594KB) ( 296 )   Objective ·To establish an in vitro culture model mimicking tumor microenvironment using hydrogel microspheres and fresh primary pancreatic cancer cells. Methods ·The morphological distribution of the hydrogel microspheres was recorded, observed and photographed under an inverted fluorescence microscope. The diameter of the microspheres was calculated by Image J, and the particle size distribution map was obtained by statistics. Renal epithelial cells (293T), pancreatic cancer cells (8988), and normal pancreatic epithelial cells (HPNE) were grown in DMEM complete medium, and passaged when the cells were 80%?90% confluent. 293T cells were cultured in DMEM medium and microsphere extract, and the proliferation curve of 293T cells cultured in the two mediums was detected by CCK-8 method to explore the biocompatibility of hydrogel microspheres. Fresh pancreatic tumor tissue was cut in the ultra-clean workbench, and pancreatic cancer tumor tissue was lysed by hyaluronidase and collagenase Ⅰ, and digested into single cell with interval shaking in a 37 ℃ water bath. The hydrogel microspheres and pancreatic cells were co-cultured in DMEM complete medium for 3 d, 1/2 of the cells were fixed with 4% paraformaldehyde, and stained with phalloidin and DAPI, and the morphology and cell distribution of the microspheres were observed under a common fluorescence microscope. The remaining 1/2 cells were used for suspension and adherent cell counting. The hydrogel microspheres and primary cells were co-cultured in the medium for 7 d, and the cell composition in the established culture model of in vitro pancreatic cancer primary cells based on the hydrogel microspheres was observed by immunofluorescence method. The pancreatic cancer tissue was embedded in paraffin, followed by paraffin tissue sectioning, hematoxylin and eosin staining of the tissue section, and observation of the pancreatic cancer tissue structure with a microscope. Results ·The size of the hydrogel microspheres was uniform, and the diameter of the microspheres was about 200 μm. The proliferation curve of 293T cells showed that the hydrogel microspheres had good biocompatibility. The co-culture experiments of hydrogel microspheres and pancreatic cell lines showed that the surface of hydrogel microspheres had strong cell affinity, which could provide support points for pancreatic cells to adhere to the surface of the microspheres and to grow normally. Through co-culture of hydrogel microspheres with fresh pancreatic cancer cells, a 3D culture model of pancreatic cancer cells in vitro was successfully established. Composition of cell types in this model was similar to that in the corresponding primary tumor tissue, which included pancreatic ductal epithelial cells, tumor stem cells, endothelial cells, fibroblasts and other cells. Conclusion ·The in vitro 3D culture model of primary pancreatic cancer cells based on hydrogel microspheres has important characteristics of pancreatic cancer tumor microenvironment. Figures and Tables | References | Related Articles | Metrics

Public health

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A prevalence study on knowledge, attitude, belief and practice of safe medication and analysis of related factors in dialysis patients WEI Shan, JI Ouyang, CHEN Zhihao, HUANG Zehui, LI Pu, FANG Junyan, LIU Yingli 2023, 43 (1):  88-94.  doi: 10.3969/j.issn.1674-8115.2023.01.011 Abstract ( 394 )   HTML ( 24 )   PDF (1347KB) ( 273 )   Objective ·To survey the status quo of knowledge, attitude, belief and practice (KAP) of safe medication in the process of medication therapy management in dialysis patients, and analyze the related factors. Methods ·This study was a cross-sectional study with the maintenance dialysis patients selected from the Department of Nephrology, Shanghai Ninth People′s Hospital, Shanghai Jiao Tong University School of Medicine from March 2019 to March 2021. The KAP scale for safe medication in patients with chronic kidney disease verified by reliability and validity tests was used to evaluate the status quo of safe medication, and their medication list and clinical characteristics were also investigated. The occurrence of medication-related problems (MRPs) was evaluated according to the Strand classification system. The related factors of KAP level of safe medication in the dialysis patients were analyzed by univariate analysis and multiple linear regression analysis. Results ·A total of 187 maintenance dialysis patients were included, including 103 patients with maintenance hemodialysis (MHD) and 84 patients with continuous ambulatory peritoneal dialysis (CAPD). The mean age of them was (60.19±12.80) years, the mean dialysis duration was (44.31±36.90) months, and the mean numbers of comorbidities and medications were 6.37±2.17 and 7.48±2.66, respectively. In the dialysis patients, the mean score of KAP scale was (77.68±18.53) points. The mean scores of knowledge, attitude, and practice of safe medication were (24.96±8.92), (17.19±3.18), and (35.40±8.18) points, respectively. The univariate analysis results showed that there were significant differences of KAP scores for safe medication in the dialysis pattern, the educational level, the economic level and the occurrence of renal anemia in the maintenance dialysis patients (all P<0.05). In addition, the more comorbidities or medications for regulating calcium and phosphorus metabolism the patients had, or the more MRPs occurred, the lower KAP scores for safe medication the patients got (all P<0.05). Multiple linear regression results showed that the dialysis patients with CAPD (β=8.391, 95%CI 3.436?13.347, P=0.001), with higher education (β=9.159, 95%CI 0.978?17.339, P=0.028), with per capita monthly household income of over 6 000 yuan (β=8.309, 95%CI 1.891?14.727, P=0.011), with less comorbidities (β=-1.582, 95%CI -2.783?-0.382, P=0.010), and with less MRPs (β=-2.284, 95%CI -3.987?-0.581, P=0.009) had higher KAP scores. Conclusion ·The KAP level of safe medication in the patients with maintenance dialysis is low. The hemodialysis, low educational level, low economic level, high number of comorbidities, and high number of MRPs are risk factors for low level of KAP of safe medication in dialysis patients. Figures and Tables | References | Related Articles | Metrics

Review

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Advances in decompensated cirrhosis treatment by human serum albumin ZHANG Chenxi, CAO Zhujun, XIANG Xiaogang, XIE Qing, GENG Jiawei 2023, 43 (1):  95-100.  doi: 10.3969/j.issn.1674-8115.2023.01.012 Abstract ( 390 )   HTML ( 34 )   PDF (1227KB) ( 465 )   Human serum albumin (HSA) is one of the most abundant proteins in the plasma which participate in plenty of physiological functions. This article reviews recent advances in HSA-related researches with respect to its synthesis, metabolism, structure, function, and clinical application in decompensated cirrhosis and its adverse events. As a multidomain polyfunctional molecule, HSA has not only shown its effect on colloid osmotic pressure elevation, but also its non-colloid functions including ligand binding capacity, antioxidant ability, immunoregulatory effect, and maintaining the stability of endothelium and permeability of capillary. However, the structure of HSA is easily affected by pathology surroundings including various posttranslational modifications of HSA, such as truncated N-terminal or C-terminal, glycosylation, and oxidation of Cys-34. Among these, the oxidation modification of Cys-34 in HSA is closely related to cirrhosis progression and has a strong prognostic ability of clinical outcomes. Besides prevention of post paracentesis circulatory dysfunction, HSA administration also shows excellent treatment potentials in the cirrhotic complications, including hepatorenal syndrome (HRS) and spontaneous peritonitis (SBP). Furthermore, more clinical trials are needed to discuss the potential benefits of HSA in non-SBP infection, long-term administration of ascites, hepatic encephalopathy, acute-on-chronic liver failure (ACLF) and other cirrhotic complications. References | Related Articles | Metrics

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Research progress in the role and mechanism of acyloxyacyl hydrolase in diseases WU Zhenkai, DENG Bo, PAN Yu, DING Feng 2023, 43 (1):  101-107.  doi: 10.3969/j.issn.1674-8115.2023.01.013 Abstract ( 488 )   HTML ( 31 )   PDF (1305KB) ( 468 )   Lipopolysaccharide (LPS) is an essential component of the outer wall of gram-negative bacteria and can induce a strong inflammatory response in the body. Acyloxyacyl hydrolase (AOAH) is a highly conserved host lipase expressed in a variety of cells, including neutrophils, macrophages and proximal tubular epithelial cells. AOAH can inactivate LPS by selectively removing the secondary acyl chains from the lipid A moiety of gram-negative bacterial LPS, which alleviates inflammation and tissue damage, promotes the recovery from the immune tolerance and regains normal responsiveness. It plays an essential role in host defenses against infection. Moreover, AOAH has been shown invivo and invitro for a variety of enzymatic activities including phospholipase, lysophospholipase and diacylglycerol lipase. Researches increasingly show that AOAH plays a critical role in various noninfectious diseases such as allergic asthma, psoriasis, chronic pelvic pain and other autoimmune diseases and chronic inflammatory diseases in addition to infections. Besides, DING Feng Research Group, Department of Nephrology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine found that AOAH was capable of attenuating renal fibrosis via proximal renal tubular epithelial cells CD74 signaling pathway. Given the increasingly diverse role of AOAH in diseases, this article reviews its sources, structure, effects and possible mechanisms in diseases in order to provide reference for future research on its roles in diseases and new lines of investigation into disease pathogenesis and novel therapeutic targets. References | Related Articles | Metrics

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Progress of residual neuromuscular blockade after general anesthesia in elderly patients LIU Qiuli, JIANG Lai, WENG Xiaojian 2023, 43 (1):  108-113.  doi: 10.3969/j.issn.1674-8115.2023.01.014 Abstract ( 415 )   HTML ( 25 )   PDF (1280KB) ( 449 )   Muscle relaxation induced by muscle relaxants is one of the important conditions for endotracheal intubation and fixation, but sometimes muscle functions recover incompletely after operations, which is called postoperative residual neuromuscular block (PRNB). It can cause a series of serious postoperative complications and even death. Hence, it is essential to prevent and treat PRNB in the early stage during anesthesia implementation. With the rapid development of surgical technology and the gradual extension of the life span of the population, the number of operations for elderly patients is gradually increasing. Meanwhile, due to the physiological and pathological changes of the elderly and the combined underlying diseases, the elderly are more likely to suffer from PRNB in different degrees after surgery, which seriously damages the prognosis of elderly patients and increases the loss of medical resources. In these years, the successful application of short-acting muscle relaxants, new antagonists (such as sugammadex), and multi-mode neuromuscular monitoring devices in clinical practice have greatly reduced the incidence of PRNB in elderly patients. Furthermore, the basic researches which mainly based on the neuromuscular junction structure and the pharmacokinetic and pharmacodynamic studies of the elderly provide a new theoretical basis for reducing the harm of PRNB to elderly patients. This paper reviews the research progress of the mechanism, adverse events and clinical interventions of PRNB in elderly patients after general anesthesia surgeries to provide reference for muscle relaxation management and postoperative antagonism in these people. References | Related Articles | Metrics

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A review of RIZ1 regulation of the signal pathways in obesity and tumors XIE Xiaolei, JIANG Peixin, ZHANG Jinghong, MO Junjian, WU Kefan, ZENG Kangyi 2023, 43 (1):  114-119.  doi: 10.3969/j.issn.1674-8115.2023.01.015 Abstract ( 354 )   HTML ( 25 )   PDF (1214KB) ( 413 )   Retinoblastoma-interacting zinc finger protein 1 (RIZ1) gene, also known as positive regulatory domain 2 (PRDM2), is a member of the PRDM gene family whose protein sequence consists of a PR domain, a nuclear hormone receptor binding motif, eight zinc finger domains, and an Rb (retinoblastoma protein) interacting motif. RIZ1 is mainly localized in the nucleus, where it plays a role in transcriptional repressor, gene regulation, protein-protein interactions, and other functions. RIZ1 is an important participant in the metabolic pathway, which affects basal metabolism and inhibits the development of obesity by regulating metabolism-related genes; functional mutations or insufficient content of RIZ1 are associated with the development of a variety of tumors, which participate in tumor processes by activating downstream oncogenes or regulating metabolism. RIZ1 regulates three molecular signal pathways, AKT (v-akt murine thymoma viral oncogene homolog)/mTOR (mechanistic target of rapamycin kinase), IGF-1 (insulin-like growth factor 1), and estrogen, in tumors and obesity through AKT3 and IGF-1, respectively, or acting as a co-activator. The functional differences of the three molecular pathways and the crossover of their downstream molecules suggest that RIZ1 may function differently in different ages, genders, and organs. The study of the regulatory role of RIZ1 and RIZ2 in metabolic processes can help to fully understand the mechanism of RIZ1 involvement in obesity and tumor formation. In the future, diagnostic research or functional recovery based on RIZ1 targets may be of great significance for the diagnosis and treatment of metabolic diseases and tumor. References | Related Articles | Metrics

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Research progress in clinical characteristics, diagnosis and prognosis of TdT-negative lymphoblastic lymphoma/acute lymphoblastic leukemia HOU Shumin, SHAO Jingbo 2023, 43 (1):  120-124.  doi: 10.3969/j.issn.1674-8115.2023.01.016 Abstract ( 683 )   HTML ( 25 )   PDF (1187KB) ( 1185 )   Terminal deoxynucleotide transferase (TdT) is a special deoxyribonucleic acid (DNA) polymerase. It can bind a single deoxynucleotide to the 3′-OH end of a DNA molecule in the absence of a template. TdT is a specific intracellular marker of immature lymphocytes in the hematopoietic system, which is of great significance in the diagnosis of lymphoid tumors. The diagnosis of lymphoblastic lymphoma/acute lymphoblastic leukemia (LBL/ALL) depends on immunotyping. TdT is usually positive in LBL/ALL, which is an important basis for clinical diagnosis of LBL/ALL. TdT-negative LBL/ALL is rare in clinical practice. Because of its strong specificity in diagnosis, negative TdT will bring certain difficulties to the diagnosis. There are many reasons for negative TdT. The expression of TdT is different in different differentiation stages of lymphocytes, and the sensitivity of different detection methods and the false negative of TdT can be seen occasionally. In addition, it has been reported that the clinical characteristics and prognosis of TdT-negative- and TdT-positive-LBL/ALL are different. Understanding the significance of TdT expression, the causes of negative TdT, and the different characteristics of TdT-negative- and TdT-positive-LBL/ALL are helpful for clinical diagnosis, treatment selection and prognosis judgment. In this paper, relevant literature at home and abroad is reviewed, and the causes of negative TdT, the clinical characteristics, diagnosis and prognosis of TdT-negative-LBL/ALL were reviewed. References | Related Articles | Metrics

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Research progress in the correlation and treatment of menopause and non-alcoholic fatty liver disease in women LIU Weiwei, WANG Long 2023, 43 (1):  125-131.  doi: 10.3969/j.issn.1674-8115.2023.01.017 Abstract ( 319 )   HTML ( 20 )   PDF (1385KB) ( 477 )   Non-alcoholic fatty liver disease (NAFLD) is a chronic non-communicable disease. It is a metabolic syndrome (MS) in the liver. Menopause is a physiological phenomenon of women due to the decline of ovarian function, which is characterized by the deterioration of hypothalamic-pituitary function. Clinical and epidemiological studies have shown that the incidence trend of NAFLD has gender differences and is related to metabolic parameters such as glucose, lipids, and blood uric acid. The incidence of NAFLD in premenopausal women is lower than that in males, while the incidence of NAFLD in postmenopausal women gradually increases to the same as that in males. The mechanism may be mainly related to the changes of sex hormones in postmenopausal women (mainly the decrease of estrogen). The changes of sex hormones such as the decreased levels of estrogen and sex hormone binding globulin (SHBG) and the relatively increased level of androgen can increase the incidence of MS components such as abdominal obesity, abnormal blood lipid metabolism and insulin resistance (IR), making menopause an important independent risk factor for NAFLD in women. Estrogen, androgen receptor antagonists and phytoestrogens can improve hepatic steatosis and IR through many ways, reduce the severity of NAFLD and delay its progression to liver fibrosis, which have certain therapeutic significance for postmenopausal women with NAFLD, but also have some limitations. This paper reviews the research progress in the relationship between menopause and NAFLD in recent years, so as to provide ideas and reference for the prevention and treatment of NAFLD in postmenopausal women. References | Related Articles | Metrics

Case report

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Monomorphic epitheliotropic intestinal T-cell lymphoma: a case with recurrent gastrointestinal hemorrhage HU Zeyu, ZHOU Cheng, YANG Lin, MA Xiaoyan, XIAO Haijuan, SI Hailong 2023, 43 (1):  132-136.  doi: 10.3969/j.issn.1674-8115.2023.01.018 Abstract ( 349 )   HTML ( 17 )   PDF (3430KB) ( 244 )   A male patient, 63-year-old, went to Xianyang Central Hospital in Shaanxi Province on September 4, 2019, for "intermittent black stool for more than 1 month with aggravation for 7 h", and was admitted as "gastrointestinal bleeding". On admission, there was fresh blood stool with a large amount, accompanied by abdominal pain, abdominal distension, dizziness, fatigue, and anorexia, without other obvious special discomfort. The electrogastroscopy indicated chronic non-atrophic gastritis with erosion; the electron colonoscopy indicated ileocecal valve inflammation; the capsule endoscopy indicated ulcer and tumor possibility. The patient was finally diagnosed as having monomorphic epitheliotropic intestinal T-cell lymphoma (MEITL) after single balloon small bowel microscopy and pathological biopsy. On November 8, 2019, the patient was transferred to the Department of Oncology, Affiliated Hospital of Shaanxi University of Chinese Medicine. After 2 cycles of chemotherapy with R-CHOP regimen (rituximab combined with cyclophosphamide, adriamycin, vincristine and prednisone), the disease progressed and then the therapy was replaced with DA-EPOCH regimen (etoposide, prednisone, vincristine, cyclophosphamide and doxorubicin). After 4 months of standardized treatment, the patient′s condition worsened again; he left hospital after giving up treatment, and died on March 20, 2020. MEITL has the characteristics of strong invasiveness, high degree of deterioration, heavy tumor load, difficult diagnosis and easy misdiagnosis, so the patients often have poor prognosis and high mortality. This article reports the clinical data of this patient with MEITL, aiming to provide reference for the diagnosis and treatment of patients with similar symptoms. Figures and Tables | References | Related Articles | Metrics