ObjectiveTo investigate the effects of targeted silencing of Runt-related Transcription Factor 3 （RUNX3） on the proliferation and migration of Mouse Hepatic Stellate Cells （HSCs）， as well as subsequent collagen deposition. MethodsMouse hepatic stellate cell line （JS-1） was selected and then morphologically observed and identified under a microscope. After the cells had fully adhered， they were treated with 5 ng/mL of transforming growth factor beta 1 （TGF-β₁） for 24 hours to induce hepatic stellate cell activation. Furthermore， a RUNX3 silencing model was established using RUNX3 lentiviral infection. The experiment was divided into four groups： Control group， TGF-β₁ group， TGF-β₁+siRNA-NC group， and TGF-β₁+siRNA-RUNX3 group. Protein expression changes of RUNX3， alpha-smooth muscle actin （α-SMA）， and Alpha 1 type I collagen （Collagen I） were detected using Western blot method. Cellular immunofluorescence assays were employed to investigate the deposition changes of α-SMA and RUNX3 in hepatic stellate cells. RT-qPCR was utilized to examine the mRNA expression changes of RUNX3， α-SMA， and Collagen I. The proliferative capacity of hepatic stellate cells was assessed using Edu staining. The migratory ability of hepatic stellate cells was evaluated through wound healing assays and Transwell migration experiments. ResultsCompared with Control group， a significant elevation in RUNX3 was observed in the TGF-β₁-induced activated HSCs （P<0.01）. Meanwhile， the protein and mRNA levels of fibrosis-related markers and α-SMA and Collagen I were significantly upregulated （P<0.001）. Additionally， the proliferation and migration capabilities of HSCs were significantly enhanced （P<0.001）. In contrast， when compared to TGF-β₁+siRNA-NC group， TGF-β₁+siRNA-RUNX3 group exhibited a notable decrease in RUNX3 and other related indicators， such as the protein and mRNA levels of α-SMA and Collagen I （P<0.05）. Concurrently， the proliferation and migration capabilities of HSCs were significantly inhibited in TGF-β₁+siRNA-RUNX3 group （P<0.01）. ConclusionSilencing RUNX3 can inhibit the deposition of collagen and the proliferation and migration of hepatic stellate cells. Conversely， RUNX3 promotes the proliferation and migration capabilities of HSCs， thereby facilitating the activation of HSC.

ObjectiveTo investigate the role of runt-related transcription factor 3 （RUNX3） in transforming growth factor-β₁ （TGF-β₁）-induced activation of mouse primary pulmonary fibroblasts （PFs）， and its effects on fibroblast activation protein （FAP） expression， cell proliferation， and collagen synthesis. MethodsPFs were isolated from C57BL/6 mice and cultured. A RUNX3 knockdown model was established using small interfering RNA （siRNA）. Cells were assigned to the control group （Control）， TGF-β₁-treated group （TGF-β₁）， negative control group （TGF-β₁+siRNA-NC）， and RUNX3-silenced group （TGF-β₁+si-RUNX3）. In addition， a RUNX3 overexpression rescue experiment was performed based on TGF-β₁ stimulation. Protein and mRNA levels of RUNX3， FAP， and typeⅠcollagen （COL1A1） were measured by Western blot and reverse transcription quantitative real-time PCR （RT-qPCR）. Cell proliferation was assessed using CCK-8 and EdU assays. Co-expression of COL1A1 and FAP was examined by double immunofluorescence staining. ResultsCompared with the Control group， RUNX3， FAP， and COL1A1 expression levels were upregulated in PFs in the TGF-β₁ group （P<0.01）. The CCK-8 assay showed that the absorbance value was reduced in the RUNX3 knockdown group compared with the negative control group （P<0.01）. Consistently， the EdU assay demonstrated a lower proportion of EdU-positive cells in the RUNX3 knockdown group than in the negative control group （P<0.01）. Immunofluorescence double staining revealed decreased fluorescence intensities of COL1A1 and FAP in the RUNX3 knockdown group relative to the negative control. Under RUNX3 overexpression conditions， these fluorescence signals exhibited a partial rebound （P<0.01）. ConclusionRUNX3 in TGF-β1-induced PFs may promote cell proliferation and collagen synthesis by positively regulating FAP expression. Targeting the RUNX3/FAP axis may represent a potential therapeutic strategy for pulmonary fibrosis.

Objective: To investigate the effect of targeted silencing of DNA methyltransferase 3A(DNMT3A) on collagen deposition, proliferation and migration activity of mouse lung fibroblasts(PFs). Methods: In order to ensure the proliferation and migration activity of primary fibroblasts, the lung tissues of neonatal C57 suckling mice were taken, PFs were extracted after being sheared, and the morphology was observed and identified under the microscope. PFs cells were activated by 5 ng/ml TGF-β1for 24 h after cell attachment, and DNMT3A silencing model was constructed by small interfering RNA; The experiment was divided into control group, TGF-β1group, TGF-β1+ siRNA-NC group and TGF-β1+ siRNA-DNMT3A group. The protein expressions of DNMT3A, α-smooth muscle actin(α-SMA) and Collagen Ⅰ were detected by Western blot; Real time quantitative reverse transcription polymerase chain reaction(RT-qPCR) was used to detect the mRNA expression changes ofDNMT3A,α-SMAandCollagenⅠ. The proliferation ability of PFs was detected by CCK-8 and EdU staining; the migration ability of PFs was detected by scratch test and Transwell migration test. Results: Compared with the control group, TGF-β1induced the increase of DNMT3A in the activated PFs cell group(P<0.01), the protein and mRNA levels of fibrosis and proliferation related indicators α-SMA and Collagen Ⅰ also increased(allP<0.05), and the proliferation and migration ability of PFs increased(allP<0.000 1). Compared with the siRNA-NC group, the protein expression levels of DNMT3A(P<0.000 1) and related indicators α-SMA(P<0.01) and Collagen Ⅰ(P<0.01) significantly decreased in the DNMT3A silencing group by Western blot, and the mRNA levels ofDNMT3A,α-SMAandCollagenⅠby RT-qPCR also decreased(allP<0.001), and the proliferation(P<0.01) and migration ability(P<0.05) of PFs cells decreased compared with the control group. Conclusion Silencing DNMT3A can inhibit the deposition of collagen and the proliferation of PFs. DNMT3A can promote the proliferation and migration of PFs, and then promote the activation of PFs and the development of pulmonary fibrosis. This process may be regulated by DNA methylation modification.

Objective To observe the inter-observer consistency of diameter of inferior vena cava(IVC)and IVC collapsibility index(IVCCI)measured and assessed with echocardiography and the correlations with right heart parameters in patients with right heart dysfunction.Methods Forty-seven patients with right heart dysfunction were prospectively recruited in observation group,while 50 adults with normal right heart function were taken as controls(control group).Parameters of the right heart were obtained with echocardiography,including the right ventricular fractional area change(FAC),the tricuspid annular plane systolic excursion(TAPSE),the myocardial performance index(MPI),the tricuspid annular systolic velocity(S')as well as early and late diastolic velocity(e',a')and e'/a' ratio,also the tricuspid valve orifice early and late diastolic velocities(E,A)and E/A ratio and E/e',the vena contracta width of tricuspid regurgitation(TR-VCW),the maximum velocity of tricuspid regurgitation(TR-Vmax),the pulmonary artery systolic pressure(PASP)and right atrial area(RAA).Besides,the maximal and minimal diameter of IVC(IVCDmax,IVCDmin)during the respiratory cycle were measured with two dimensional(2D)ultrasound and anatomical M-mode ultrasound,respectively,and the IVCCI were calculated.Then 20 subjects were randomly selected from each group,and IVC parameters were obtained.The basic data,right heart parameters and IVC parameters were compared between groups,intra-class correlation coefficient(ICC)between 2 sonographers of IVC parameters were calculated,and correlations between IVC parameters and right heart parameters were assessed.Results No significant differences of gender,age nor body mass index(BMI)was detected between groups(all P＞0.05).Compared with those in control group,MPI,e',e'/a',E,A,E/e',TR-VCW,TR-Vmax,PASP and RAA increased,whereas FAC,TAPSE,S'and a'decreased in observation group(all P＜0.05).The inter-observer consistencies were good for IVCDmax and IVCCI in observation group(ICC=0.787-0.971)and IVCDmax in the control group(ICC=0.971,0.964)obtained with 2D ultrasound and anatomical M-mode ultrasound,but poor for IVCCI in control group(ICC=0.169,0.456).Compared with those in control group,IVC parameters 2D-IVCDmax,2D-IVCDmin,M-IVCDmax and M-IVCDmin increased but 2D-IVCCI and M-IVCCI decreased in observation group(all P＜0.05).In control group,2D-IVCDmax was weakly negatively correlated with TAPSE and a'(r=-0.392,-0.364),weakly positively correlated with e'/a',E,E/A,TR-VCW and RAA(r=0.396,0.483,0.461,0.565,0.582),2D-IVCCI was weakly negatively correlated with TR-VCW and RAA(r=-0.386,-0.380),while M-IVCDmax was weakly negatively correlated with TAPSE(r=-0.384),and weakly positively correlated with e'/a',E,E/A,TR-VCW and RAA(r=0.357,0.453,0.473,0.549,0.550),M-IVCCI was weakly negatively correlated with MPI,E,TR-VCW and RAA(r=-0.347,-0.337,-0.475,-0.421).Conclusion In patients with right heart dysfunction,IVCD diameter and IVCCI obtained with echocardiography had good inter-observer consistencies.Parameters obtained with 2D ultrasound and anatomic M-mode ultrasound had certain relations with the right heart parameters.

Balloon-assisted enteroscopy(BAE)technology includes double-balloon enteroscopy(DBE)and single-balloon enteroscopy(SBE),and plays a significant role in the diagnosis and treatment of small intestinal diseases such as small intestinal bleeding,Crohn's disease,small intestinal tumors,and incomplete small intestinal obstruction.With technological advancements,BAE has not only improved diagnostic rates but also achieved minimally invasive treatment.Its safety and efficacy have been widely recognized,and it is expected to play a broader role in the diagnosis and treatment of small intestinal diseases in the future.

Balloon-assisted enteroscopy(BAE)technology includes double-balloon enteroscopy(DBE)and single-balloon enteroscopy(SBE),and plays a significant role in the diagnosis and treatment of small intestinal diseases such as small intestinal bleeding,Crohn's disease,small intestinal tumors,and incomplete small intestinal obstruction.With technological advancements,BAE has not only improved diagnostic rates but also achieved minimally invasive treatment.Its safety and efficacy have been widely recognized,and it is expected to play a broader role in the diagnosis and treatment of small intestinal diseases in the future.

Objective:To evaluate the in vitro cross-neutralization of serum antibodies in human and mice immunized with inactivated SARS-CoV-2 vaccine against Delta and Beta variants. Methods:Human serum samples after a second and a third dose of inactivated SARS-CoV-2 vaccine and mouse serum samples after a two-dose vaccination were collected. The neutralizing antibodies in the samples against SARS-CoV-2 strains of prototype, Delta and Beta variants were detected using micro-neutralization assay in biosafety level Ⅲ laboratory. The seroconversion rates and geometric mean titers (GMTs) of antibodies were calculated.Results:The seroconversion rates of antibodies in human serum samples against different SARS-CoV-2 strains were all above 95%. After two-dose vaccination, the GMTs of neutralizing antibodies against the prototype, Delta and Beta strains were 109, 41 and 15, respectively. The GMTs decreased by 2.7 folds and 7.3 folds for the Delta and Beta variants as compared with the prototype strain. After the booster vaccination, the GMTs of neutralizing antibodies against the prototype, Delta and Beta strains were 446, 190 and 86, respectively. The GMTs of neutralizing antibodies against Delta and Beta variants decreased by 2.3 folds and 5.2 folds as compared with that against the prototype strain. The seroconversion rates of antibodies against different SARS-CoV-2 strains in mouse serum samples were all 100%. The GMTs of neutralizing antibodies against the prototype, Delta and Beta strains were 2 037, 862 and 408, respectively. The GMTs decreased by 2.4 folds and 5.0 folds for the Delta and Beta variants.Conclusions:Inactivated SARS-CoV-2 vaccine could induce a certain level of neutralizing antibodies against Delta and Beta variants in both human and mouse models. Moreover, a third dose of vaccine induced higher levels of neutralizing antibodies against Delta and Beta variants in human. This study provided valuable data for the clinical application and protective evaluation of the inactivated SARS-CoV-2 vaccine.

Objective:To establish the prediction model of postoperative Gleason score (GS) risk of prostatic cancer (PCa), and to compare the diagnostic efficacy of the model and each independent risk factor for PCa medium-high risk group.Methods:The clinical data of 362 patients who accepted transrectal prostate biopsy in the Run Run Shaw Hospital Affiliated to Zhejiang University School of Medicine from January 2018 to December 2019 were analyzed retrospectively, and a total of 343 patients with prostate cancer who met the enrollment criteria were selected. According to the GS grading system, these patients were divided into low risk group, moderate risk group and high risk group. At first, the single factor analysis and Spearman rank correlation were used to find out the effective indicators with good correlation with GS risk. Then, multiple linear regression equation was applied for multi-factor analysis to obtain the independent risk factors and the prediction model for predicting GS risk, and then the ROC curve was used to compare the diagnostic efficacy of each independent risk factor and prediction model for PCa medium-high risk group.Results:In the single factor analysis, the differences of all indicators in GS risk were statistically significant (all P<0.05). In the correlation analysis with GS risk, except for the indicators of prostate volume (all P>0.05), the other indexes had linear correlations with the different risks of GS (all P<0.05). Among them, the total prostate specific antigen and two-dimensional ultrasound (2D-US) score showed moderate positive correlations( rs=0.402, 0.579, all P<0.001), contrast enhanced ultrasound (CEUS) score showed a high positive correlation ( rs=0.709, P<0.001), and the rest indexes showed low positive correlations. Multiple linear regression was used to obtain two independent risk factors of 2D-US score ( X1) and CEUS score ( X2) for the prediction of GS risk, then, a prediction model was established: Y=0.863+ 0.066 X1+ 0.27 X2, the corresponding linear coefficient differences were statistically significant(all P<0.05). By the ROC analysis, the areas under the curves of 2D-US score, CEUS score and the prediction model were 0.838, 0.906 and 0.907, respectively. Conclusions:2D-US score and CEUS score are independent risk factors for predicting postoperative GS risk, and the diagnostic efficacy of the prediction model is higher than those of the 2D-US score and CEUS score for the medium-high risk group.

Objective: To understand the change of smoking status among middle school students in Quzhou City from 2012 to 2017, and to provide a basis for school tobacco control. Methods: Through stratified cluster sampling method, Zhejiang youth health-related behavior questionnaire survey was conducted among 1 454 students in May 2012 and among 1 655 students in May 2017. Results: Among middle school students in Quzhou, attempting smoking rate, ever-smoking a whole cigarette reporting rate and current-smoking report rate decreased from 31.36%, 17.81% and 8.73% in 2012 to 21.70%,12.45% and 6.22% in 2018 respectively, with statistically significant differences(χ2=37.30,17.52,7.12,P<0.01). Among the current-smoking students, the rate of smoking frequency≥20 d/month and heavy-smoking rate decreased from 30.71% and 7.09% in 2012 to 15.53% and 0.97% respectively. It showed that attempting smoking rate, ever-smoking a whole cigarette reporting rate and current-smoking report rate of male students were higher than that of female students(χ2=117.50,63.99;47.14,51.26;110.18,22.50), and the non-ordinary high school students were higher than that of ordinary high school students and middle school students across two waves surveillance(χ2=47.14,51.26;70.34,43.77;62.67,22.50)(P<0.01). Conclusion School tobacco control have made a great progress，the tobacco use behavior of middle school students declines sharply in Quzhou.

Objective: Non-suicidal self-injury (NSSI) in adolescents appeared prevalent and multifactorial. This study was to examine the associations between exposure to suicidal behaviors and NSSI in the Chinese adolescents. Methods: Participants included for analyses were 5 154 adolescent students who participated in the baseline survey in 2015 and the first follow-up survey in 2016 of the Shandong Adolescent Behavior and Health Cohort, but with no history of NSSI at the baseline survey. A self-administered structured questionnaire was used to collect data on demographics, behavioral and emotional problems, lifetime and last-year NSSI. Data on the history of exposure to suicide attempt or death of a family member, friend, or close acquaintance were also collected. Multivariate logistic regression methods were used to examine the associations between exposure to suicidal behaviors and NSSI. Results: In the baseline survey, mean age of the 5 154 participants was (14.49±1.48) years, with 48.5% of the participants as girls. Of the participants, 9.0% reported having been exposed to suicidal behaviors, including 6.0% reported to suicide attempt, 4.9% to suicide death, 7.3% to suicidal behaviors of friends/close acquaintances, and 3.1% to suicidal behaviors of relatives. The prevalence rates of NSSI in the last year were significantly higher in adolescents who had been exposed to suicidal behaviors than those who had not (P<0.05). Results from the multivariate logistic regressions showed that exposure to suicide death (OR=1.91, 95%CI: 1.22-3.01) or to suicidal behaviors of relatives (OR=1.79, 95%CI: 1.02-3.12) were both significantly associated with the increased risk of NSSI. Conclusions Experiences related to exposure to suicide-death or suicidal behaviors of relatives were associated with increased risk of NSSI in adolescents. After the suicide events, psychological counseling and health education programs set for high-risk groups were helpful in promoting physical and mental health and preventing the attempt of self-injury in teenagers.