Objective To investigate the efficiency and mechanism of differentiation of reprogrammed adipose-derived stem cells (ADSCs) to neurons in vitro.Methods ADSCs from rats were cultured in vitro and then purified and identified.ADSCs at the third passage were divided into three groups:ADSCs without lentivirus-mediated gene transfection (blank group),ADSCs transfected with lentivirus carrying no neurogenin2 (Ngn2) (empty virus group) and ADSCs with lentivirus-mediated transfection of Ngn2 (Ngn2 group).All groups were induced in the medium containing cell growth factor for 15 days.The positive expression of neuron-specific nuclear protein (NeuN) in three groups was detected using immunofluorescence method so as to observe the efficiency of neuron differentiation.Expression variances of Mash1,Hes1 and Dll1 in each group were detected by Western blot analysis and the mechanism of differentiation was also discussed.Results After 15 days of induction,positive expression rate of NeuN in Ngn2 group,empty virus group,blank group was 90.12％,45.34％ and 40.26％ respectively,with significant differences among groups (P ＜ 0.01).Western blot analysis showed that Ngn2 group had a significantly higher expression of Dll1 (P ＜0.01) and obvious lower expressions of Hes1 and Mash1 (P ＜0.01),as compared with empty virus group and blank group.However,there were no significant differences of expression levels of Dll1,Mash1 and Hes1 between empty virus group and blank group (P ＞ 0.05).Conclusions After induction,the ratio of neuron differentiation of reprogrammed ADSCs is increased by almost 99％,as compared with simple ADSCs.The increased dfferentiation of reprogrammedADSCs to neurons may be associated with the inhibition of notch signaling through up-regulating Dll1 and down-regulating Mash1 and Hesl.

Objective To establish ulcerative colitis(UC)model in BALB/c mice and to investigate the role of high mobility group box chromosomal protein 1(HMGBI)in pathogenesis of UC.Methods Thirty-two BALB/c mice were randomly divided into UC group(n=24,which were fed with 3%dextran sulfate 80dium solution)and control group(n=8,which were fed with water).The animals were sacrificed at 24.96 and 1 68 hours,respectively,to collect samples of colon and blood.The sernm level of HMGB1 was measured with ELISA and the expression of HMGB1 in colon was determined by Western blotting analysis.Results Histological scoring increased with the induction of the model,and manifestation of colon mucosa at 168h was similar with that of UC in human.The serum level of HMGB1 was slightly higer at 24 h than that of control(5.09±0.61 μg/L vs 4.49±0.53μg/L,P＞0.05),and reached a peak at 96 h (14.74±0.60 μg/L,P＜0.01),decreased at 168 h(9.03±0.78μg/L,P＜0.01).The expression 0.05).significandy increased at 96h(0.76±0.03,P＜0.05)and at 168 h(0.77±0.04,P＜0.05).Conclusion HMGB1 might be involved in pathologic changes of UC at a later stage.

Helicobacter pylori (Hp) infection is closely related to gastric cancer, and with the implementation of eradication therapy, the infection rate of Hp decreases gradually and the proportion of gastric cancer caused by Hp infection also decreases accordingly. Hence, Helicobacter pylori-negative gastric cancer (HpNGC) become a more common type of gastric cancer. It has been reported in recent years that there are some differences in pathobiological behavior between Helicobacter pylori-positive gastric cancer (HpPGC) and HpNGC, which indicates that the pathogenesis of HpNGC may be different from that of HpPGC. Relevant studies have found that the expression profile of microRNAs (miRNAs) in HpNGC is different from that in HpPGC, suggesting that the differentially expressed miRNAs may play an important role in the pathogenesis of HpNGC. This article reviewed the research progress on miRNAs in the pathogenesis, diagnosis and treatment of HpNGC, providing a new insight for exploring the pathogenesis of HpNGC.

Background and purpose:Rabs are members of Ras-related small GTPase superfamily. Rab27A is a unique member in the Rab family and has specific implications in human genetic diseases. We studied the potential role of Rab27A in proliferation, distribution of cell cycle, apoptosis and invasion of breast cancer cells and its mechanism(s). Methods:The eukaryotic expression vector containing Rab27A open reading frame (ORF) pcDNA3.1(+) - Rab27A was constructed and transfected into MDA-MB-231 breast cancer cells. Then we detected the changes in terms of cell growth, cell cycle distribution, apoptosis and in vitro invasion capability before and after transfection. We also applied RT-PCR to investigate the molecular basis.Results:① The expression of Rab27A was increased as invasive and metastatic ability increased in four human breast cancer cell lines. ② Overexpression of Rab27A can promote breast cancer cells to grow faster, increase the proportion of S phase cells, avoid apoptosis and invade in vitro. ③ Rab27A transfectants constitutively enhanced the expression of Cyclin D1, MMP-7 and MMP-9 in MDA-MB-231 cell lines, on the contrary, that of p16 were down-regulated constitutively. Reduced Rab27A expression by RNAi down-regulated the expression of Cyclin D1, MMP-7 and MMP-9, and up-regulated p16 expression.Conclusions:Rab27A can stimulate breast cancer cells to proliferate, increase the proportion of cells in S phase,avoid apoptosis and invade in vitro by regulating the expression of Cyclin D1, MMP-7, MMP-9 and p16.

Objective:To analyze the relationship between gait speed or grip strength and all-cause mortality in elderly inpatients over 75 years old, and to compare their predictive value for all-cause mortality.Methods:A prospective cohort study was conducted and enrolled elderly patients aged ≥75 years hospitalized from December 2016 to December 2019 at the Department of Integrated Medicine and Geriatrics, Fuxing Hospital, Capital Medical University.Gait speed(m/s)and grip strength(kg)were respectively measured via the 6-meter walk test and a dynamometer.The patients were followed up for more than 1 year after discharge, and the time of all-cause mortality was recorded.The Cox regression model was used to analyze the correlation between gait speed, grip strength or their combination and the risk of all-cause mortality.ROC curves were statistically analyzed using the DeLong test.Results:A total of 704 patients were enrolled, with an average age of(83.8±6.3)years; the median follow-up time was 33(24, 42)months.During the follow-up period, all-cause death occurred in 131 cases(18.6%).Compared with the high gait speed and high grip strength groups, the low gait speed and low grip strength groups had higher all-cause mortality(all P<0.05).The Cox regression model was used to analyze the relationships between gait speed, grip strength and all-cause mortality.The results showed that gait speed( HR=2.255, 95% CI: 1.462-3.477, P<0.001)and grip strength( HR=1.815, 95% CI: 1.232-2.673, P<0.001)were associated with the risk of all-cause mortality after adjustment for other factors; When gait speed slowed down and grip strength decreased, the risk of death reached the highest level( HR=3.156, 95% CI: 1.829-5.445, P<0.001).The AUC of the gait speed model(0.703, 95% CI: 0.667-0.736)was higher than the AUC of the grip strength model(0.648, 95% CI: 0.611-0.683), with a difference of 0.055(95% CI: 0.006-0.103, P=0.026). Conclusions:Decreased gait speed or grip strength is related to an increase of death risk.The risk of death is highest when the patient has both slowed gait speed and decreased grip strength.The predictive value of gait speed for death is better than grip strength.Together they can be used as simple, rapid and effective tools to predict all-cause mortality in this population.

Objective To investigate the targeting of transforming growth factor β1(TGF-β1)expression by Ran-binding protein 9(RANBP9)and its effect on colorectal cancer Colo320 cell apoptosis.Methods Gene expression levels of RANBP9 were analyzed in 625 colon cancer tissues and 20 normal colon tissues in The Cancer Genome Atlas database.The relationship between RANBP9 expression and survival time of patients with colon cancer was analyzed using KMPLOT.The expression of TGF-β1 in normal colon tissues and colon cancer tissues was analyzed using the human protein immunohistochemistry database and the relationship between TGF-β1 expression and the survival of patients with colon cancer was analyzed using the UALCAN database.The relationship between RANBP9 and TGF-β1 was analyzed by dual luciferase experiments.Colo320 cells were transfected with pcDNA3.1-GFP-RANBP9 and control pcDNA3.1-GFP-RANBP9-NC plasmids,respectively,and normal control cells were established without transfection.The cells were cultured and the growth viability of each group of cells was detected by the iazolyl blue tetrazolium bromide method,apoptosis was detected by flow cytometry,the mitochondrial membrane potential was detected by JC-1 staining,and RANBP9 and TGF-β1 protein expression were detected by Western blot.Results RANBP9 expression was significantly reduced in colon cancer tissues.Compared with patients with low RANBP9 expression,patients with high RANBP9 expression had a higher survival curve and significantly higher expression of TGF-β1 in colon cancer tissue.Compared with patients with high TGF-β1 expression,patients with low TGF-β1 expression had a significantly higher survival curve(P＜0.05).RANBP9 targeted the expression of TGF-β1 in colon cancer.Compared with the normal group,cell growth,mitochondrial membrane potential,and TGF-β1 expression were all significantly down-regulated and the apoptosis rate and RANBP9 expression were significantly increased in the experiment group(P＜0.05).Conclusions RANBP9 can target the expression of TGF-β1,promote the growth of Colo320 colon cancer cells,decrease the mitochondrial membrane potential,and induce apoptosis.

Objective:To compare the efficacy between deep continuous irrigation combined with vacuum sealing drainage (VSD) and routine dressing change in treating multidrug-resistant bacterial infections at the surgical wound site in patients with major vascular injury.Methods:A retrospective cohort study was conducted to analyze the clinical data of 28 patients with surgical wound infections by multidrug-resistant bacteria after major vascular injury treated at the First Affiliated Hospital of Henan University of Science and Technology from March 2015 to December 2021. There were 15 males and 13 females, aged 15-65 years [(41.8±12.9)years]. All patients received vascular graft surgery after major vascular injury. Postoperative microbiological culture indicated that the wound infections were caused by Carbapenem-resistant organisms (CRO) or vancomycin- resistant Enterococci (VRE), with no available sensitive antibiotics for treatment. The patients received surgical debridement every five days after vascular graft surgery and were divided into two groups to receive the subsequent treatments including a routine dressing change (routine dressing group, 14 patients) or a deep continuous irrigation combined with VSD (irrigation combined with VSD group, 14 patients). On the first day post-operation and then every 3 days, inflammatory indicators [white blood cell count, neutrophils, C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), and procalcitonin] were observed in the two groups (repeat tests when a patient′s condition changed). Microbiological cultures were applied with patient samples every 5 days to observe the wound and infection control. Comparisons were made between the two groups regarding the duration to normal levels of inflammatory indicators, duration to negative CRO or VRE cultures, visual analogue score (VAS) before and at 1, 2 and 3 hours after changing the irrigation fluid (changing the dressing), conditions of wound skin grafting or flap repair, and incidences of anastomotic fistula.Results:All patients were followed up for 12-24 months [(14.3±2.4)months], during which no wound redness, rupture, purulent discharge or infection recurrence was noted. The duration to normal levels was (9.4±2.4)days for white blood cells, (9.6±2.8)days for neutrophils, (9.8±3.1)days for CRP, (12.2±3.6)days for ESR, and (7.6±1.9)days for procalcitonin in the irrigation combined with VSD group, significantly shorter than those in the routine dressing group [(15.2±3.1)days, (13.6±3.4)days, (14.2±3.9)days, (19.9±3.3)days, and (12.9±4.1)days, respectively] (all P<0.01). The duration to negative CRO or VRE cultures was (13.9±3.1)days in the irrigation combined with VSD group, significantly shorter than that in the routine dressing group [(19.2±6.9)days] ( P<0.05). The VAS before and at 1, 2 and 3 hours after changing the irrigation fluid was (4.2±0.7)points, (4.1±0.9)points, (4.2±0.9)points and (4.1±0.8)points in the irrigation combined with VSD group, respectively, and was (4.3±0.6)points, (6.9±0.7)points, (5.4±0.9)points and (4.5±0.9)points in the routine dressing group, respectively. The VAS score in the irrigation combined with VSD group was significantly lower than that in the routine dressing group at 1 hour and 2 hours after changing the irrigation fluid (all P<0.01), while no significant differences were found before and at 3 hours after changing the irrigation fluid (all P>0.05). After infection control, 5 patients (35.7%) in the irrigation combined with VSD group required skin grafting or flap repair at the wound site, lower than 11 patients (78.6%) in the routine dressing group ( P<0.01). The incidence of anastomotic fistula was 7.1% (1/14) in the irrigation combined with VSD group, lower than 42.9% (6/14) in the routine dressing group ( P<0.05). Conclusion:When multidrug-resistant bacterial infections occur at the surgical wound site after major vascular injury, deep continuous irrigation combined with VSD performs better than routine dressing change in controlling infection as well as in reducing pain, rate of wound skin grafting or flap repair and incidence of anastomotic fistula, without reliance on antibiotics.

Objective: Analysis of the effect of triggering receptor-1 expressed on myeloid cells (TREM-1) in nonalcoholic fatty liver disease (NAFLD) and the mechanism. Methods: The oleic acid-treated HepG2 cells were divided into model group, overexpression group, interference group A, interference group B and negative control group. The mouse model of NAFLD was generated and randomly divided into (nuclear factor-κB) NF-κB inhibition group, protein kinase B (AKT) inhibition group, knockout group A, knockout group B and control group. The expression of inflammatory factors and TREM-1 in liver tissue was detected by PCR, and fat accumulation was detected by oil red O staining. Western blotting was used to detect the expression of TREM-1 and signaling pathway proteins, and HE staining was used to detect liver tissue changes. Results: TREM-1 was up-regulated in liver tissue of NAFLD mice [(0.936±0.127) vs. (0.432±0.105)] and in oleic acid-treated HepG2 cells. In oleic acid-treated HepG2 cells, overexpression of TREM-1 increased inflammatory factor expression and increased lipid droplets; inhibition of TREM-1 expression decreased inflammatory factor expression, and lipid droplets decreased. Knockout of TREM-1 and inhibition of NF-κB in NAFLD mice reduced hepatocyte inflammatory factor expression and reduced liver damage; knockout of TREM-1 and inhibition of AKT reduced liver tissue lipids and drops accumulate. Conclusions The overexpression of TREM-1 in NAFLD mice liver tissue can regulate inflammatory factor expression and lipid droplets through NF-κB and AKT signal pathway. TREM-1 might be a potential therapeutic target of NAFLD.