Objective:To investigate the expressions of cystathionine-β-synthase(CBS)and cystathionine-γ-lyase(CSE)and their effects on the malignant biological behaviours of breast cancer cells,and to elucidate their mechanisms.Methods:The breast cancer tissue and paracancerous normal tissue from 15 cases of patients were selected,and RT-qPCR and Western blotting methods were used to detect the mRNA and protein expression levels of CBS and CSE in breast cancer tissue,paracancerous normal tissue,MCF-7 cells,and MDA-MB-231 cells.The MCF-7 cells were divided into siNC group(transfected with siNC)and siCBS group(transfected with siCBS),and the MDA-MB-231 cells were divided into ovNC group(transfected with CSE over-expression empty plasmid)and ovCSE group(transfected with CSE over-expression plasmid).CCK8 assay was used to detect the proliferation activities of breast cancer cells in various groups,Transwell assay was used to detect the numbers of migration and invasion cells in various groups,and Western blotting method was used to detect the protein expression levels of E-cadherin,N-cadherin and Vimentin proteins in the breast cancer cells in various groups.Results:Compared with paracancerous normal tissue,the expression levels of CBS and CSE mRNA and proteins in breast cancer tissue were increased(P＜0.05 or P＜0.01).Compared with MDA-MB-231 cells,the CBS mRNA expression level in the MCF-7 cells was increased(P＜0.05);compared with MCF-7 cells,the expression level of CSE protein in the MDA-MB-231 cells was decreased(P＜0.05).Compared with siNC group,the proliferation activity,the numbers of migration and invasion cells,the expression levels of N-cadherin and Vimentin proteins in the MCF-7 cells in siCBS group were significantly decreased(P＜0.05),and the expression level of E-cadherin protein was increased(P＜0.05).Compared with ovNC group,the proliferation activity,the numbers of migratoin and invasion cells,and the expression levels of N-cadherin and Vimentin proteins in the MDA-MB-231 cells in ovCSE group were increased(P＜0.05),while the expression level of E-cadherin protein was significantly decreased(P＜0.05).Conclusion:The expressions of CBS and CSE are upregulated in breast cancer tissue,and high levels of CBS and CSE promote proliferation,migration,invasion and epithelial-mesenchymal transition(EMT)of breast cancer cells.

Objective To investigate the diagnostic value of serum bromodomain-containing pro-tein 4(BRD4)combined with gastrin-17(G-17)in Helicobacter pylori(Hp)-positive early gastric cancer.Methods A total of 88 patients with Hp-positive early gastric cancer admitted to our hospital from September 2021 to September 2023 were selected as early gastric cancer group.Meanwhile,92 patients with Hp-positive precancerous lesions and 80 patients with Hp-positive gastritis admitted dur-ing the same period were selected as precancerous lesion group and gastritis group,respectively.En-zyme-linked immunosorbent assay(ELISA)was used to detect the serum levels of BRD4 and G-17.Multivariate logistic regression analysis was performed to screen the influencing factors of Hp-positive early gastric cancer.Receiver operating characteristic(ROC)curve analysis was used to evaluate the diagnostic value of serum BRD4 and G-17 in Hp-positive early gastric cancer.Results Compared with the gastritis group,the levels of BRD4 and G-17 in the precancerous lesion group and early gastric cancer group were significantly increased(P＜0.05).Furthermore,compared with the precancerous le-sion group,the levels of BRD4 and G-17 in the early gastric cancer group were also significantly elevated(P＜0.05).The proportion of patients with a family history of gastric cancer,those who preferred hot food,cold food,or high-salt food,as well as the levels of PG Ⅱ,BRD4,and G-17 were signifi-cantly higher in the early gastric cancer group than in the non-gastric cancer group,while the level of pepsinogen(PG)Ⅰ was significantly lower(P＜0.05).Logistic regression analysis revealed that preference for hot food,high-salt food,PG Ⅱ,BRD4,G-17,and PG Ⅰ were all influencing factors for Hp-positive early gastric cancer(P＜0.05).The area under the curve(AUC)values for serum BRD4,G-17,and their combination in diagnosing Hp-positive early gastric cancer were 0.793,0.830,and 0.912,respectively.The diagnostic efficacy of the combined detection was superior to that of single detection(P＜0.05).Conclusion The serum levels of BRD4 and G-17 are elevated in patients with Hp-positive early gastric cancer,and both exhibit certain diagnostic value for Hp-positive early gastric cancer,suggesting their potential as serum biomarkers for the diagnosis of Hp-positive early gastric cancer.

Objective To investigate the role of knockdown of peroxiredoxin-6(PRDX6)in injury and adaptive expression of bile acid transporter in human hepatoellular carcinomas(HepG2)cells induced by rifampicin(RFP).Methods Cells in logarithmic growth phase were uniformly inoculated in six-well plates,and HepG2 cells were transiently transfected with specific PRDX6-siRNA and control-siRNA to construct the knockdown group and control group.After 24 h of induction with 100 μmol/L RFP,Western blot and qRT-PCR were performed to detect the protein and gene expression levels of PRDX6,multidrug resistance protein 1(MDR1),multidrug resist-ance-associated proteins 2,3 and 4(MRP2,MRP3 and MRP4),and Na+/taurine taurocholate cotransporter pro-tein(NTCP).Annexin V-FITC/PI double staining assay was used to detect the apoptosis rate of cells in each group;CCK-8 assay was used to detect the changes of cell proliferation in each group;The relative contents of ala-nine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBIL),indirect bilirubin(IBIL)and total bile acid(TBA)in the supernatant of cell culture medium of each group were detected by kits.Results RFP increased the protein and gene expression levels of MRP2,MRP3,MRP4,MDR1,NTCP and PRDX6 in HepG2 cells(P<0.05),while the protein and gene expression levels of MRP2,MRP3,MRP4,MDR1 and NTCP decreased to different degrees after PRDX6 knockdown(P<0.05).In addition,PRDX6 knockdown re-sulted in increased apoptosis rate of HepG2 cells(P<0.05),decreased cell proliferation ability(P<0.05),and increased levels of cell injury markers(ALT,AST,TBIL,DBIL,TBA)in cell culture supernatants(P<0.05).Conclusion RFP increased the protein and gene expression of bile acid transporter and PRDX6 to increase in HepG2 cells.However,following knockdown of PRDX6 and treatment with RFP,the protein and gene expression levels of the bile acid transporter decreased and cell injury was aggravated,suggesting that PRDX6 played a protec-tive role in RFP-induced adaptive response in HepG2 cells.

Osteoarthritis(OA),characterized by cartilage degeneration,synovial inflammation,and subchondral bone remodeling,is among the most common musculoskeletal disorders globally in people over 60 years of age.The initiation and progression of OA involves the abnormal metabolism of chondrocytes as an important pathogenic process.Cartilage degeneration features mitochondrial dysfunction as one of the important causative factors of abnormal chondrocyte metabolism.Therefore,maintaining mitochondrial homeostasis is an important strategy to mitigate OA.Mitophagy is a vital process for autophagosomes to target,engulf,and remove damaged and dysfunctional mitochondria,thereby maintaining mitochondrial homeostasis.Cumulative studies have revealed a strong association between mitophagy and OA,suggesting that the regulation of mitophagy may be a novel therapeutic direction for OA.By reviewing the literature on mitophagy and OA published in recent years,this paper elaborates the potential mechanism of mitophagy regulating OA,thus providing a theoretical basis for studies related to mitophagy to develop new treatment options for OA.

The endoplasmic reticulum is a key site for protein production and quality control.More than one-third of proteins are synthesized and folded into the correct three-dimensional conformation in the endoplasmic reticulum.However,during protein folding,unfolded and/or misfolded proteins are prone to occur,which may lead to endoplasmic reticulum stress.Organisms can monitor the quality of the proteins produced by endoplasmic reticulum quality control(ERQC)and endoplasmic reticulum-associated degradation(ERAD),which maintain endoplasmic reticulum protein homeostasis by degrading abnormally folded proteins.The underlying mechanisms of protein folding and ERAD in mammals have not yet been fully explored.Therefore,this paper reviews the process and function of protein folding and ERAD in mammalian cells,in order to help clinicians better understand the mechanism of ERAD and to provide a scientific reference for the treatment of diseases caused by abnormal ERAD.

Enterotoxigenic Escherichia coli（ETEC）infection can induce watery diarrhea，leading to dehydration，electrolyte disturbance，and even death in severe cases. Recombinant B subunit/inactivated whole-cell cholera（rBS/WC）vaccine is effective in preventing ETEC infectious diarrhea. On the basis of the latest evidence on etiology and epidemiology of ETEC，as well as the effectiveness，safety，and health economics of rBS/WC vaccine，National Clinical Research Center for Child Health（The Children’s Hospital，Zhejiang University School of Medicine）and Zhejiang Provincial Center for Disease Control and Prevention invited experts to develop expert consensus on rBS/WC vaccine in prevention of ETEC infectious diarrhea. It aims to provide the clinicians and vaccination professionals with guidelines on using rBS/WC vaccine to reduce the incidence of ETEC infectious diarrhea.

Objective : To investigate the effect of mesencephalic astrocyte-derived neurotrophic factor (MANF) on the adaptive expression of bile acid transporter in human hepatoellular carcinomas (HepG2) induced by rifampicin (RFP) ． Methods: The control group cell line (Y07) and the knockdown group cell line (Y25) were constructed by lentiviral stable transfection technology．The Y07 and Y25 cells were treated with RFP of 200 μmol / L for 48 h， and qRT-PCR and Western blot were used to detect the protein and gene expression levels of MANF，bile salt export pump ( BSEP) ，multidrug resistance-related proteins 2 /3 /4 ( MRP2 ，MRP3 ，MRP4) ，multidrug resistance protein 1 (MDR1) ，organic solute transporter a / β ( OSTα/ β) ，organic anion transporter ( OATP2B1) ．The protein and gene expression levels of proliferating cell nuclear antigen ( PCNA) ，proliferating cell marker Ki67 were used to evaluate the proliferation of cells in each group changes in levels．Changes in the protein and gene expression levels of C / EBP homologous protein( CHOP) and cysteinyl aspartate specific proteinase-3 ( Caspase-3) were used to evaluate the apoptosis of cells in each group．The relative contents of alanine aminotransferase(ALT) ，aspartate aminotransferase(AST) ，alkaline phosphatase ( ALP) ，total bilirubin ( TBIL) ，indirect bilirubin ( IBIL) and total bile acid(TBA) in the supernatant of cell culture medium of each group were detected by kits. Results: RFP could induce the protein and gene expression of MANF，BSEP ，MRP2 ，MRP3 ，MRP4 ，MDR1 ，OSTα , OSTβ , OATP2B1 in HepG2 cells (P ＜0. 05 ) ，while the protein and gene expression levels of BSEP ，MRP2 ，MRP3， MRP4，MDR1，OSTα、OSTβ、OATP2B1 decreased after MANF knockdown(P＜0. 05) ．Moreover，under the action of RFP，the protein expression of PCNA and Ki67 in the knockdown group was still higher．The protein and gene levels of CHOP and Caspase-3 significantly increased after MANF knockdown(P＜0. 05) ．The levels of the hepatic cell injury markers in the cell supernatant increased significantly(P＜0. 05) ． Conclusion RFP can induce the expression of bile acid transporter such as BSEP，MRP2，MRP3，MRP4，MDR1，OSTα , OSTβ and OATP2B1 to increase in HepG2 cells(P＜0. 05) ，but the expression of bile acid transporter of HepG2 after MANF knockdown will significantly decrease under the induction of rifampicin(P＜0. 05) ，and cell indury is aggravated，indicating that MANF plays a protective role in RFP-induced adaptive responses by regulating the bile acid transporter.

Objective: To detect the difference in the incidence of small intestinal bacterial overgrowth(SIBO) in patients with liver cirrhosis and chronic hepatitis fibrosis(CHF) with the lactulose hydrogen breath test(LHBT), and to explore the relationship between SIBO and inflammatory factors and oxidative stress related indicators. Methods: LHBT was performed on 38 CHF patients, 60 cirrhosis patients and 31 healthy controls to evaluate the incidence of SIBO. The patients were further divided into SIBO-positive and negative group. Then we compared related clinical symptoms and laboratory tests between the two groups and detected lipopolysaccharide(LPS), interleukin(IL)-6, tumor necrosis factor(TNF)-α, IL-10 level, and several kinds of oxidative stress indicators such as diamine oxidase(DAO), superoxide dismutase(SOD), glutathione(GSH), catalase(CAT). Statistical analysis was conducted to analyze the correlation between the concentration of LPS, IL-6, TNF-α, DAO, SOD, GSH, CAT and LHBT summation value. Results: (1) The positive rate of SIBO in CHF group, cirrhosis group and control group was 36.84%, 60.00% and 9.68%, respectively. The difference was statistically significant(P<0.01).(2) The differences in CTP classification and ascites were statistically significant between the two subgroups with and without SIBO(P<0.05).(3) The levels of LPS, IL-6, CAT, DAO and SOD in SIBO-positive group were higher than those in negative group(P<0.05). However, the concentrations of IL-10, TNF-α and GSH were similar between the two groups.(4) The LHBT summation value was positively correlated with the concentrations of LPS, IL-6, DAO, SOD and CAT in serum(P<0.05), but had no significant correlation with TNF-α, IL-10 and GSH. Conclusion Compared with healthy controls, CHF and cirrhosis patients are more likely to develop SIBO. It is also related to the increase of inflammatory factors and oxidative stress indexes in peripheral blood. SIBO may aggravate the inflammatory response of CHF and cirrhosis patients through intestinal flora dysregulation and oxidative stress, thus aggravating the disease change.

Objective:To investigate the efficacy of floating acupuncture therapy in the treatment of chronic refractory pressure ulcers and its effects on serum levels of superoxide dismutase, malondialdehyde and insulin-like growth factor-1.Methods:A total of 60 patients with chronic refractory pressure ulcers who received treatment in The Second Hospital of Jiaxing from October 2019 to October 2021 were included in this study. They were randomly divided into observation and control groups, with 30 patients in each group. The patients in the control group were given routine treatment, while the patients in the observation group were given floating acupuncture therapy based on routine treatment. Clinical efficacy, wound healing rate after 7, 14 and 21 days of treatment, the time of emerging granulation tissue, the time of shedding rotten meat, and the time of wound healing, changes in serum levels of superoxide dismutase, malondialdehyde and insulin-like growth factor-1 after 21 days of treatment relative to those before treatment were compared between the two groups.Results:Total response rate in the observation group was significantly higher than that in the control group [90.0% (27/30) vs. 66.7% (20/30), χ2 = 4.81, P < 0.05]. Wound healing rate after 7, 14 and 21 days of treatment in the observation group was significantly higher than that in the control group ( t = -5.45, -7.77, -7.51, all P < 0.001). The time of emerging granulation tissue, the time of shedding rotten meat, and the time of wound healing in the observation group were significantly shorter than those in the control group ( t = 5.69, 9.26, 8.91, all P < 0.001). After 21 days of treatment, serum levels of superoxide dismutase, malondialdehyde and insulin-like growth factor-1 in the observation group were (96.52 ± 10.43) U/mL, (4.32 ± 0.78) μmol/L, (43.52 ± 8.29) μg/L, respectively, which were significantly different from (83.76 ± 6.81) U/mL, (5.48 ± 0.92) μmol/L, (36.78 ± 9.18) ng/mL, in the control group ( t = 5.61, 5.27, 2.98, all P < 0.05). Conclusion:Floating acupuncture therapy is effective in the treatment of refractory pressure ulcer. It can accelerate wound healing, increase serum superoxide dismutase and insulin-like growth factor-1 levels, and decrease serum malondialdehyde level.

Objective:To explore any effect of training assisted by a pelvic rehabilitation robot on trunk control and walking after cerebral infarction.Methods:Forty cerebral infarction survivors with hemiplegia were randomly divided into an experimental group and a control group, each of 20. Both groups were given routine neurological medication and rehabilitation training, while the experimental group was additionally provided with 20 minutes of robot-assisted gait training daily, five times a week, for 4 weeks. Before and after the intervention, the motor function, walking function, trunk control and pelvic movement were assessed using a simplified version of the Fugl-Meyer assessment (FMA-LL), functional ambulation categories (FAC) and the trunk control test (TCT).Results:After the treatment, significant improvement was observed in all of the above measurements in both groups. The average FMA-LL, FAC and TCT results of the experimental group as well as their average pelvic lateral displacement, height displacement, rotation angle and roll angle were all significantly superior to the control group′s averages.Conclusions:Robot-assisted training can effectively improve lower limb motor functioning, trunk control, walking and pelvic motion after cerebral infarction, with better curative effect than routine rehabilitation training alone.