OBJECTIVES: To evaluate the regulatory effects of lncRNA LINC00261 on proliferation, invasion, and metastasis of esophageal squamous cell carcinoma (ESCC) cells. METHODS: The differentially expressed RNAs in ESCC were identified using the GSE149612 dataset from the GEO database. PCR was used to detect LINC00261 expression levels in clinical ESCC and normal esophageal tissue samples and in multiple ESCC cell lines and normal esophageal epithelial cells (HEEC). In ESCC cells, the effects of overexpression of LINC00261 on cell proliferation, invasion, metastasis and apoptosis were analyzed using CCK-8 assay, clone formation assay, Transwell assay and flow cytometry. The potential targets of LINC00261 were predicted using bioinformatics tools including ENCORI and verified using dual-luciferase reporter assay and Western blotting. The effects of LINC00261 overexpression on ESCC were confirmed in a nude mouse model bearing ESCC xenograft. RESULTS: Analysis of the GSE149612 dataset revealed significantly lower LINC00261 expression in ESCC tissues and cell lines. In cultured ESCC cells, LINC00261 overexpression markedly suppressed cell proliferation, invasion, and metastasis and promoted cell apoptosis. Dual-luciferase reporter assays confirmed that LINC00261 targets the miR-23a-3p/ZNF292 axis. In the tumor-bearing mouse model, LINC00261 overexpression significantly inhibited ESCC xenograft proliferation and metastasis. CONCLUSIONS LINC00261 suppresses ESCC progression by targeting the miR-23a-3p/ZNF292 axis, suggesting a potential therapeutic strategy for ESCC treatment.
Humans ; MicroRNAs/genetics* ; Cell Proliferation ; Esophageal Neoplasms/genetics* ; Animals ; Esophageal Squamous Cell Carcinoma ; Mice, Nude ; RNA, Long Noncoding/genetics* ; Cell Line, Tumor ; Neoplasm Invasiveness ; Mice ; Carcinoma, Squamous Cell/genetics* ; Apoptosis ; Gene Expression Regulation, Neoplastic ; Neoplasm Metastasis

Objective:To investigate the association of exposure to PM 2.5 and its constituents during pregnancy and fetal growth and to further identify critical windows of exposure for fetal growth. Methods:We included 4 089 mother-child pairs from the Jiangsu Birth Cohort Study between January 2016 and October 2019. Data of general characteristics, clinical information, daily average PM 2.5 exposure, and its constituents during pregnancy were collected. Fetal growth parameters, including head circumference (HC), abdominal circumference (AC), and femur length (FL), were measured by ultrasound after 20 weeks of gestation, and then estimated fetal weight (EFW) was calculated. Generalized linear mixed models were adopted to examine the associations of prenatal exposure to PM 2.5 and its constituents with fetal growth. Distributed lag nonlinear models were used to identify critical exposure windows for each outcome. Results:A 10 μg/m 3 increase in PM 2.5 exposure during pregnancy was associated with a decrease of 0.025 ( β=-0.025, 95% CI: -0.048- -0.001) in HC Z-score, 0.026 ( β=-0.026, 95% CI: -0.049- -0.003) in AC Z-score, and 0.028 ( β=-0.028, 95% CI:-0.052--0.004) in EFW Z-score, along with an increased risk of 8.5% ( RR=1.085, 95% CI: 1.010-1.165) and 13.5% ( RR=1.135, 95% CI: 1.016-1.268) for undergrowth of HC and EFW, respectively. Regarding PM 2.5 constituents, prenatal exposure to black carbon, organic matter, nitrate, sulfate (SO 42-) and ammonium consistently correlated with decreased HC Z-score. SO 42- exposure was also associated with decreased FL Z-scores. In addition, we found that gestational weeks 2-5 were critical windows for HC, weeks 4-13 and 19-40 for AC, weeks 4-13 and 23-37 for FL, and weeks 4-12 and 20-40 for EFW. Conclusions:Our findings demonstrated that exposure to PM 2.5 and its constituents during pregnancy could adversely affect fetal growth and the critical windows for different fetal growth parameters are not completely consistent.

Objective:To discuss the effect of Fuzheng Ruanjian Anticancer Formula on the malignant biological behaviors of the hepatocellular carcinoma HepG2 cells by requlating protein kinase B(Akt)/murine double minute 2(MDM2)/P53 signaling pathway.Methods:The HepG2 cells were treated with 0,0.05,0.10,0.20,0.40,0.80,1.60,3.20,and 6.40 g·mL-1 Fuzheng Ruanjian Anticancer Formula for 48 h.CCK-8 method was used to detect the survival rates of the HepG2 cells in various groups,and the concentrations of Fuzheng Ruanjian Anticancer Formula for the subsequent experiments were screened.The HepG2 cells were divided into control group,low dose of Fuzheng Ruanjian Anticancer Formula group(0.2 g·mL-1),medium dose of Fuzheng Ruanjian Anticancer Formula group(0.4 g·mL-1),high dose of Fuzheng Ruanjian Anticancer Formula group(0.8 g·mL-1),SC79 group(8 mg·L-1 SC79),and high dose of Fuzheng Ruanjian Anticancer Formula+SC79 group(0.8 g·mL-1 Fuzheng Ruijian Anticancer Formula+8 mg·L-1 SC79).CCK-8 method was used to detect the proliferation activities of the HepG2 cells in various groups;clone formation assay was used to detect the clone formation rates of the HepG2 cells in various groups;flow cytometry was used to detect the apoptotic rates of the HepG2 cells in various groups;Transwell chamber assay was used to detect the numbers of migration and invasion HepG2 cells in various groups;Western blotting method was used to detect the expression levels of proliferating cell nuclear antigen(PCNA),cysteine aspartate specific proteinase(Caspase-3),matrix metalloproteinase(MMP)-2,MMP-9,phosphorylated Akt(p-Akt),phosphorylated MDM2(p-MDM2),and P53 proteins in the HepG2 cells in various groups.Results:As the increasing of concentrations of Fuzheng Ruanjian Anticancer Formula(0,0.05,0.10,0.20,0.40,0.80,1.60,3.20,and 6.40 g·mL-1),the surival rates of the HepG2 cells were gradually decreased(P<0.05),and 0.2,0.4,and 0.8 g·mL-1 Fuzheng Ruanjian Anticancer Formula were selected for the subsequent experiments.The CCK-8 assay results showed that compared with control group,the proliferation activities of the HepG2 cells in low,medium,and high doses of Fuzheng Ruanjian Anticancer Formula groups were significantly decreased(P<0.05),in a dose-dependent manner,while the proliferation activity of the cells in SC79 group was significantly increased(P<0.05).Compared with high dose of Fuzheng Ruanjian Anticancer Formula group,the proliferation activity of the HepG2 cells in high dose of Fuzheng Ruanjian Anticancer Formula+SC79 group was significantly increased(P<0.05).The clone formation assay results showed that compared with control group,the clone formation rates of the HepG2 cells in low,medium,and high doses of Fuzheng Ruanjian Anticancer Formula groups were significantly decreased(P<0.05)in a dose-dependent manner,while the clone formation rate of the cells in SC79 group was significantly increased(P<0.05);compared with high dose of Fuzheng Ruanjian Anticancer Formula group,the clone formation rate of the cells in high dose of Fuzheng Ruanjian Anticancer Formula+SC79 group was significantly increased(P<0.05).The flow cytometry results showed that compared with control group,the apoptotic rates of the HepG2 cells in low,medium,and high doses of Fuzheng Ruijian Anticancer Formula groups were significantly increased(P<0.05)in a dose-dependent manner,while the apoptotic rate of the cells in SC79 group was significantly decreased(P<0.05);compared with high dose of Fuzheng Ruanjian Anticancer Formula group,the apoptotic rate of the cells in high dose of Fuzheng Ruanjian Anticancer Formula+SC79 group was significantly decreased(P<0.05).The Transwell chamber assay results showed that compared with control group,the numbers of migration and invasion HepG2 cells in low,medium,and high doses of Fuzheng Ruanjian Anticancer Formula groups were significantly decreased(P<0.05)in a dose-dependent manner,while the numbers of migration and invasion cells in SC79 group were significantly increased(P<0.05);compared with high dose of Fuzheng Ruanjian Anticancer Formula group,the numbers of migration and invasion HepG2 cells in high dose of Fuzheng Ruanjian Anticancer Formula+SC79 group were significantly increased(P<0.05).The Western blotting results showed that compared with control group,the expression levels of PCNA,MMP-2,MMP-9,p-Akt,and p-MDM2 proteins in the cells in low,medium,and high doses of Fuzheng Ruanjian Anticancer Formula groups were significantly decreased(P<0.05)in a dose-dependent manner,while the expression levels of Caspase-3 and P53 proteins were significantly increased(P<0.05)in a dose-dependent manner,while the expression levels of PCNA,MMP-2,MMP-9,p-Akt,and p-MDM2 proteins in the cells in SC79 group were significantly increased(P<0.05),and the expression levels of Caspase-3 and P53 proteins were significantly decreased(P<0.05);compared with high dose of Fuzheng Ruanjian Anticancer Formula group,the expression levels of PCNA,MMP-2,MMP-9,p-Akt,and p-MDM2 proteins in the cells in high dose of Fuzheng Ruanjian Anticancer Formula+SC79 group were significantly increased(P<0.05),while the expression levels of Caspase-3 and P53 proteins were significantly decreased(P<0.05).Conclusion:Fuzheng Ruanjian Anticancer Formula may inhibit the proliferation,migration,and invasion of the HepG2 cells and promote the apoptosis,and its mechanism may be related to suppressing the Akt/MDM2 signaling pathway and upregulating the P53 proteim expression.

Objective: To analyze the frequency and composition of risk-related cytogenetic abnormalities (CAs) in patients with newly-diagnosed multiple myeloma (NDMM) . Methods: The frequency and composition of risk-related CAs from a cohort of 1 015 Chinese patients with NDMM were determined by interphase fluorescence in situ hybridization (iFISH) , individually or in combination. Results: Of the cohort of 1 015 Chinese patients with NDMM, the frequencies of IgH arrangement, del (13q) /13q14, 1q gain and del (17p) were 54.0%, 46.4%, 46.1% (35.8% and 12. 7% for 3 or more than 3 copies) and 9.9%, respectively. Among 454 patients who had the baseline information for all risk-related CAs [except t (14;20) , which was not covered by the FISH panels performed routinely at all five centers], the frequencies of t (4;14) , t (11;14) or t (14;20) were 14.1%, 11.2% and 4.8%, respectively; of them, 44.3% patients carried 2 or more CAs (28.0%, 13.4% and 2.9% for 2, 3 or ≥4 CAs) ; 83.3%, 95.0% or 68.6% patients with 1q gain, del (17p) or IgH rearrangement had 1 or more additional CA (s) , with del (13q) /13q14 as the most frequently accompanied CA; 57.7% patients carried at least 1 HRCA; the incidences of double-hit (DH) MM (DHMM) (=2 HRCAs) and triple-hit (TH) (THMM) (≥3 HRCAs) were 14.3% and 2.9%, respectively. Conclusions Our results provided an up-to-date profile of CAs in Chinese NDMM patients, which revealed that approximately 58% patients might carry at least 1 HRCA, and 17% could experience so-called DHMM or THMM who presumably had the worst outcome.

Objective:To investigate the effect of sodium arsenite (NaAsO 2) on autophagy protein microtubule-associated protein light chain 3 (LC3) and Beclin-1 in human hepatic stellate cells (LX-2 cells). Methods:LX-2 cells were cultured in vitro, and LX-2 cells were stably infected with red fluorescent protein-green fluorescent protein-microtubule-LC3 (RFP-GFP-LC3) lentivirus. Flow cytometry was used for screening and infection rate determination. Using a group design, different concentrations NaAsO 2 [μmol/L: 5.00 (infection + high arsenic dose group), 0.50 (infection + medium arsenic dose group), 0.05 (infection + low arsenic dose group), and 0.00 (infection group)] were incubated to stably infect LX-2 cells, and an in vitro liver fibrosis model was constructed, and a blank group was established. The effect of NaAsO 2 on the activity of LX-2 cells was detected by CCK-8 method. The mRNA and protein expression levels of LC3, Beclin-1 and α-smooth muscle actin (α-SMA) were detected by real-time fluorescent quantitative PCR and Western blotting. Results:After stable infection of LX-2 cells by RFP-GFP-LC3 lentivirus, the fluorescence rate of RFP and GFP was determined by flow cytometry, and the infection rate was about 70%. There was no significant difference in the fluorescence intensity of RFP and GFP observed under fluorescence microscope, and the infected cells were established successfully. After treatment with NaAsO 2 for 24, 48, 72 h, compared with the blank group, the cell viability of the infection group was not significantly different statistically ( P > 0.05), and the cell viability of other dose groups decreased ( P < 0.05). There were significant differences in the expression levels of LC3, Beclin-1, α-SMA mRNA and protein in blank, infection, infection + high arsenic dose, infection + medium arsenic dose, and infection + low arsenic dose groups ( F = 17.450, 11.084, 11.294, 11.745, 31.635, 12.130, P < 0.05). In infection, infection + high arsenic dose, and infection + low arsenic dose groups, the levels of LC3 mRNA (20.09 ± 6.50, 36.57 ± 9.68, 14.19 ± 6.17) were higher than that of the blank group (1.25 ± 0.21, P < 0.05), and the level of LC3 mRNA in infection + high arsenic dose group was higher than that of the infection group ( P < 0.05); in infection, infection + high arsenic dose, infection + medium arsenic dose, and infection + low arsenic dose groups, the levels of Beclin-1 mRNA (22.46 ± 0.66, 13.38 ± 2.27, 20.80 ± 6.95, 24.31 ± 7.09) were higher than that of the blank group (1.10 ± 0.53, P < 0.05); in infection + high arsenic dose, infection + medium arsenic dose, and infection + low arsenic dose groups, the levels of α-SMA mRNA (1.07 ± 0.27, 1.65 ± 0.17, 1.73 ± 0.26) were higher than that of the blank and infection groups (0.60 ± 0.11, 0.31 ± 0.09, P < 0.05). In infection, infection + medium arsenic dose, and infection + low arsenic dose groups, the LC3 protein expressions (0.20 ± 0.06, 0.15 ± 0.00, 0.16 ± 0.01) were significantly increased compared to that of the blank group (0.04 ± 0.01, P < 0.05); in infection + medium arsenic dose, and infection + low arsenic dose groups (0.83 ± 0.03, 1.20 ± 0.02), the Beclin-1 protein expressions were significantly increased compared to that of the blank group (0.25 ± 0.01, P < 0.05), and the Beclin-1 protein expression in infection + low arsenic dose group was increased compared to that of the infection group (0.53 ± 0.03, P < 0.05); in infection + medium arsenic dose, and infection + low arsenic dose groups (0.78 ± 0.10, 0.68 ± 0.06), the α-SMA protein expressions were significantly increased compared to that of the blank and infection groups (0.40 ± 0.07, 0.48 ± 0.04, P < 0.05). Conclusion:NaAsO 2 may affect the process of arsenic-induced liver fibrosis by promoting the autophagy level of LX-2 cells.

Objective: To investigate the associated factors and the independent risk factors for portal vein thrombosis (PVT) in cirrhotic patients and assess the influences of anticoagulation on esophagogastric variceal bleeding (EGVB) in these patients. Methods: From January 2012 to December 2012, 239 cirrhotic patients were diagnosed in our hospital. According to the presence or absence of portal vein thrombosis (PVT), they were divided into thrombus group (33 cases) and control group (206 cases). According to the presence or absence of EGVB in thrombus group, they were divided into bleeding group (10 cases) and non bleeding group (23 cases). According to whether anticoagulant therapy was used in thrombus group, they were divided into anticoagulant group (10 cases) and non anticoagulant group (23 cases). The risk factors of each group and its control group were observed and compared. Results: The thrombus group had a lower level of the albumin (ALB) , higher level of count of platelet (PLT), diameter of main portal vein (MPV), propotion of diabetes prevalence and history of splenectomy compared with the control group (P<0.01). According to unconditional logistic regression analysis, both the PLT and the diameter of MPV were identified as independent risk factors for PVT in cirrhotic patients (P=0.009, 0.001; OR=1.006, 16.858). There were significant differences in the degree of varicose veins and the proportion of sequential endoscopic treatment between the bleeding group and the control group (P<0.05). Moreover, the group treated with anticoagulant drugs and the group without anticoagulation were followed up and observed for 1 year, which showed no significant changes in the bleeding ratio between two groups [40% (4/10) versus 26.1% (6/23), P>0.05]. Conclusions ⑴ PLT, ALB, MPV, and a history of diabetes or splenectomy are risk factors for cirrhosis combined with PVT, and PLT and MPV are independent risk factors. ⑵ The incidence of EGVB increased with the increasing severity of esophagogastric varicose vein. The endoscopic variceal sequential treatment can contribute a significant reduction of EGVB in cirrhosis complicated by PVT. ⑶ Anticoagulant therapy may not raise the incidence of EGVB in cirrhotic patients with PVT.

Objective To explore the value of primary and Advanced levels of laparoscopic simulating training in different seniority of gynecologic residents.Methods 77 residents in their first to forth training-year were divided into two groups:1-2 year resident and 3-4 year resident,trained with different levels of simulating training plans respectively and then assessed in the Department of Obstetrics and Gynecology in Peking University Third Hospital.Results The qualified rate of primary and advanced simulation training was 85.7％ and 57.1％ respectively.The qualified rate of primary training (80.4％ vs.100.0％,P=0.028) and advanced training (12.5％ vs.57.14％,P=0.000) were significantly different between 1-2 year residents and 3-4 year residents.The operative skills improved significantly in all the residents.In the 1-2 year residents,the scores of the primary training increased more obviously,while in the 3-4 year residents,the scores of the advanced training increased significantly.Conclusion It might be more effective for residents with different seniority to receive different levels of simulating training accordingly,so as to improve their laparoscopic operative skills more effectively.

Objective: To establish a rat model of polycystic ovary syndrome(PCOS) complicated with atherosclerosis(AS). Methods: Sixteen female SD rats were selected and randomly divided into control group and model group, with 8 rats in each group.The rats in the control group were given routine rearing.The rats in the model group were subcutaneously given dihydrotestosterone(DHT) in neck and fed with high fat diet for a long term.The changes of food intake (FI), body weight(BW), testosterone (T), estrogen (E₂), luteinizing hormone (LH), total cholesterol (TC), blood glucose (BG) and insulin (INS) were observed in the two groups. Results: The levels of FI, T, E₂, LH between the control group and model group had no obvious change[(86.13±7.83)g/r vs.(96.25±10.66)g/r, t=2.113, P=0.563, (10.79±1.74)mg/L vs.(11.47±1.89)mg/L, t=1.785, P=0.087; (36.58±2.57)ng/L vs.(38.64±1.78)ng/L, t=2.697, P=0.068; (15.47±1.96)IU/L vs.(16.01±0.80)IU/L, t=1.570, P=0.614]. The levels of BW, TC, DHT, INS in the model group were significantly higher than those in the control group[(234.54±17.14)g vs.(192.67±16.47)g, t=7.930, P<0.000; (2.47±0.13)mmol/L vs.(2.02±0.15)mmol/L, t=6.475, P<0.000; (139.75±12.12)ng/L vs.(55.63±7.80)ng/L, t=8.697, P<0.000; (283.25±33.47)pg/mg vs.(162.12±15.51)pg/mg, t=9.289, P<0.000]. Tube wall was markedly thickened in the model group after being given high fat feed for 12 weeks, and intimal wall was significantly thickened after 16 weeks, and endotheliocyte injury, deep collagen fiber, monocyte adhesion, visible atherosclerotic plaques were observed in the model grouop. Conclusion DHT-induced PCOS rat model by high fat feed reproduces the human typical clinical symptoms and pathological characteristics, it can provide a relatively simple and feasible rat model for further study of the mechanism of PCOS complicated with AS.

Objective To analyze the overactive bladder symptom score (OABSS) in evaluating bladder dysfunction in the different stages among type 2 diabetes mellitus patients,and to explore the value of bladder hyperactivity symptom score in screening early diabetic bladder dysfunction.Methods A total of 1 157 patients with type 2 diabetes mellitus,aged 40-88 yearswith mean age of 60.2 years,were enrolled from October 2013 to October 2016.The survey included the patients' characteristics,past history,current history,OABSS and quality of life (QOL) index scores.T test,single factor analysis of variance and multiple regression analysis are used to analyze the results.Results As many as 1 157 were qualified for final statistical analysis.OABSS is 1.94 ± 1.23 in group with diabetes duration ＜ 10 years,3.24 ± 1.45in group with diabetes duration 10-20 years,and 4.00 ± 1.72 in group with diabetes duration ＞ 20 years.The differences of OABSS in the different duration of diabetes was statistically significant.As diabetic duration increased,OABSS value increased (F =48.419,P ＜ 0.001).The difference of OABSS in the different HbA1c level,age and concurrent peripheral neuropathy was statistically significant.There was no significant difference of OABSS in the different BMI and distinct therapies.There was no significant difference of OABSS in diabetes with hypertension and without hypertension,with cardiopathy and without cardiopathy,with cerebrovascular disease (CVD) and without CVD,with hyperhpemia and without hyperlipemia.The significant factors were used to make multivariate analysis.The results showed that the duration of diabetes,HbA1 c level,age,peripheral neuropathy were still statistically significant.Standardized partial regression coefficient of diabetic duration was 0.366.OABSS was positively correlated with QOL score (r =0.434,P ＜ 0.001).Conclusions The related symptoms in OABSS with diabetic bladder dysfunction is correlated with the duration of diabetes,HbA1c level,age,concurrent peripheral neuropathy among type 2 diabetes.The duration of diabetes was the most significant factor.OABSS is likely to be one of the tools to assess the early symptoms of diabetic bladder dysfunction.

OBJECTIVE:To establish a method for the 5 main components (original syringin,chlorogenic acid,eleutheroside E,isofraxidin and quercetin-3-rhamnoside) in the fruits and roots of wild Acanthopanax senticosus. METHODS:UPLC was per-formed on the column of Waters ACQUITY UPLC HSS T3 with mobile phase of acetonitrile-0.3% phosphoric acid (gradient elu-tion)at a flow rate of 0.2 ml/min. Detection wavelength was 300 nm,column temperature was 30 ℃,and injection volume was 10μl. RESULTS:The linear range was 24.56-184.2 μg/ml for syringin(r=0.9993),18.454-138.405 μg/ml for chlorogenic acid(r=0.9993),8.416-63.12 μg/ml for eleutheroside E (r=0.9997),3.286-24.645 μg/ml for isofraxidin (r=0.9993) and 2.522-18.915μg/ml for quercetin-3-rhamnoside(r=0.9998);RSDs of precision,stability and reproducibility tests were lower than 1%;recover-ies were 99.14%-100.50%(RSD=0.48%,n=6)for syringing in the fruits of A. senticosus、99.03%-100.45%(RSD=0.50%,n=6) for chlorogenic acid in the fruits of A. senticosus、99.22%-100.44%(RSD=0.44%,n=6)for eleutheroside E in the fruits of A. sen-ticosus、99.80%-100.80%(RDS=0.44%,n=6)for isofraxidin in the fruits of A. senticosus、99.76%-101.10%(RSD=0.51%,n=6) for quercetin-3-rhamnoside in the fruits of A. senticosus;99.21%-101.20%(RSD=0.73%,n=6)for syringing in the root of A. senti-cosus、99.81%-101.20%(RSD=0.52%,n=6)for chlorogenic acid in the root of A. senticosus、100.00%-101.50%(RSD=0.62%, n=6)for eleutheroside E in the root of A. senticosus、99.22%-100.40%(RSD=0.47%,n=6)for isofraxidin in the root of A. senti-cosus. CONCLUSIONS:The method is simple and stable with good reproducibility,and can be used for the simultaneous determi-nation of original syringin,chlorogenic acid,eleutheroside E,isofraxidin and quercetin-3-rhamnoside in the fruits and root of wild A. senticosus.