Objective:To investigate the role of cyclooxygenase-2 (COX-2) in glycochenodeoxycholic acid (GCDC)-induced apoptosis of mouse extrahepatic biliary epithelial cells (EBECs) and clarify its possible mechanism.Methods:EBECs were cultured in vitro and infected with RNAi-COX-2 lentivirus (GCDC+ shCOX-2 group). EBECs were then treated with different concentrations (0, 25, 50, 100, 200, 400, 800 μmol/L) of GCDC (GCDC group). Cell proliferation activity was detected by cell counting kit-8 (CCK-8) assay; lactate dehydrogenase (LDH) release rate was measured by colorimetry; apoptosis was analyzed by flow cytometry; caspase-3 activity was detected by fluorescent probe method; COX-2 mRNA expression was determined by real-time quantitative polymerase chain reaction (qRT-PCR); protein levels of COX-2, autophagy-related proteins LC3, p62, Beclin-1, and apoptosis-related proteins Bax, Bcl-2, cleaved caspase-3 were evaluated by Western blot.Results:Compared with the 0 μmol/L GCDC group, the apoptosis level, LDH release rate, and caspase-3 activity in the 50, 100, and 200 μmol/L GCDC groups gradually increased (all P<0.05) in a concentration-dependent manner. After RNAi-COX-2 lentivirus infection, COX-2 mRNA and protein expression levels in EBECs significantly decreased (all P<0.05). Compared with the GCDC group, the apoptosis rate of EBECs in the GCDC+ shCOX-2 group significantly decreased ( P<0.05). The GCDC+ shCOX-2 group showed increased cell proliferation activity, downregulated protein expressions of Bax, cleaved caspase-3, and p62, and upregulated protein expressions of Bcl-2, LC3 II/I, and Beclin-1 (all P<0.05). Conclusions:Inhibition of COX-2 improves GCDC-induced apoptosis of EBECs, and the mechanism may be related to the activation of the autophagy pathway.

Objective:To investigate the role of cyclooxygenase-2 (COX-2) in glycochenodeoxycholic acid (GCDC)-induced apoptosis of mouse extrahepatic biliary epithelial cells (EBECs) and clarify its possible mechanism.Methods:EBECs were cultured in vitro and infected with RNAi-COX-2 lentivirus (GCDC+ shCOX-2 group). EBECs were then treated with different concentrations (0, 25, 50, 100, 200, 400, 800 μmol/L) of GCDC (GCDC group). Cell proliferation activity was detected by cell counting kit-8 (CCK-8) assay; lactate dehydrogenase (LDH) release rate was measured by colorimetry; apoptosis was analyzed by flow cytometry; caspase-3 activity was detected by fluorescent probe method; COX-2 mRNA expression was determined by real-time quantitative polymerase chain reaction (qRT-PCR); protein levels of COX-2, autophagy-related proteins LC3, p62, Beclin-1, and apoptosis-related proteins Bax, Bcl-2, cleaved caspase-3 were evaluated by Western blot.Results:Compared with the 0 μmol/L GCDC group, the apoptosis level, LDH release rate, and caspase-3 activity in the 50, 100, and 200 μmol/L GCDC groups gradually increased (all P<0.05) in a concentration-dependent manner. After RNAi-COX-2 lentivirus infection, COX-2 mRNA and protein expression levels in EBECs significantly decreased (all P<0.05). Compared with the GCDC group, the apoptosis rate of EBECs in the GCDC+ shCOX-2 group significantly decreased ( P<0.05). The GCDC+ shCOX-2 group showed increased cell proliferation activity, downregulated protein expressions of Bax, cleaved caspase-3, and p62, and upregulated protein expressions of Bcl-2, LC3 II/I, and Beclin-1 (all P<0.05). Conclusions:Inhibition of COX-2 improves GCDC-induced apoptosis of EBECs, and the mechanism may be related to the activation of the autophagy pathway.

OBJECTIVE To investigate the inhibitory effects of Bruceine A(BA)on colon cancer and its underlying mechanisms.METHODS Human colon cancer HT-29 and HCT116 cells were treated with various concentrations of BA(0,1,2,5,10,20,40,80 μmol·L-1).Cell viability was assessed using the Cell Counting Kit-8(CCK-8).Flow cytometry,wound healing assays,and Transwell assays were employed to evaluate the effects of BA on cell apoptosis,cell cycle,invasion,and migration.Mo-lecular docking simulations were used to assess the binding of BA to GSK-3β protein,and Western blot analysis was used to examine protein expression related to the cell cycle,epithelial-mesenchymal transition,and the Wnt/β-catenin signaling pathway.An HT-29 cell subcutaneous xenograft mouse model was established.After tumor formation,mice were randomly divided into three groups(six mice per group):a blank group,a low-dose BA group(0.1 mg·kg-1),and a high-dose BA group(0.2 mg·kg-1).Mice were ad-ministered the drug for 19 d,then sacrificed,and tumor tissues were collected.Tumor volume changes over time were observed;Ki67 immunohistochemistry was used to assess cell proliferation in tumor tissues;Western blot analysis of Wnt/β-catenin signaling pathway protein expression was conducted.RESULTS Compared with the blank group,BA could significantly inhibit the proliferation of HT-29 and HCT116 cells,with IC50 values of 10.80 μmol·L-1 and 17.96 μmol·L-1,respectively.Flow cytometry results showed that BA significantly induced apoptosis of HT-29 cells(P<0.01,P<0.001),and arrested the cell cycle at the S phase,accompanied by de-creased expression of cycle-related proteins CDK2 and Cyclin A(P<0.05,P<0.01,P<0.001).BA inhibited cell migration and in-vasion ability(P<0.05,P<0.01,P<0.001),reduced the expression of EMT-related proteins Snail,Vimentin,and N-Cadherin(P<0.01,P<0.001),and upregulated the expression of E-Cadherin protein.In addition,BA inhibited the expression of β-catenin and p-GSK3β proteins.Wnt agonist LiCl could significantly antagonize the anti-colon cancer effect of BA;Wnt inhibitor XAV939 could enhance the anti-colon cancer effect of BA.In the in vivo experiment,compared with the blank group,the tumor volume of the low-dose and high-dose BA groups was significantly reduced(P<0.05,P<0.001).Immunohistochemistry results showed that compared with the blank group,the expression of Ki67 in tumor tissues of the low-dose and high-dose BA groups was significantly reduced(P<0.001).Western blot results further proved that BA inhibited the Wnt/β-catenin signaling pathway.CONCLUSION BA inhibits the viability,invasion,and migration of colon cancer HT-29 cells,induces apoptosis,and causes cell cycle arrest.Additionally,it significantly suppresses the growth of subcutaneous HT-29 cell xenografts in vivo,possibly related to the Wnt/β-catenin signaling pathway.

OBJECTIVE To investigate the inhibitory effects of Bruceine A(BA)on colon cancer and its underlying mechanisms.METHODS Human colon cancer HT-29 and HCT116 cells were treated with various concentrations of BA(0,1,2,5,10,20,40,80 μmol·L-1).Cell viability was assessed using the Cell Counting Kit-8(CCK-8).Flow cytometry,wound healing assays,and Transwell assays were employed to evaluate the effects of BA on cell apoptosis,cell cycle,invasion,and migration.Mo-lecular docking simulations were used to assess the binding of BA to GSK-3β protein,and Western blot analysis was used to examine protein expression related to the cell cycle,epithelial-mesenchymal transition,and the Wnt/β-catenin signaling pathway.An HT-29 cell subcutaneous xenograft mouse model was established.After tumor formation,mice were randomly divided into three groups(six mice per group):a blank group,a low-dose BA group(0.1 mg·kg-1),and a high-dose BA group(0.2 mg·kg-1).Mice were ad-ministered the drug for 19 d,then sacrificed,and tumor tissues were collected.Tumor volume changes over time were observed;Ki67 immunohistochemistry was used to assess cell proliferation in tumor tissues;Western blot analysis of Wnt/β-catenin signaling pathway protein expression was conducted.RESULTS Compared with the blank group,BA could significantly inhibit the proliferation of HT-29 and HCT116 cells,with IC50 values of 10.80 μmol·L-1 and 17.96 μmol·L-1,respectively.Flow cytometry results showed that BA significantly induced apoptosis of HT-29 cells(P<0.01,P<0.001),and arrested the cell cycle at the S phase,accompanied by de-creased expression of cycle-related proteins CDK2 and Cyclin A(P<0.05,P<0.01,P<0.001).BA inhibited cell migration and in-vasion ability(P<0.05,P<0.01,P<0.001),reduced the expression of EMT-related proteins Snail,Vimentin,and N-Cadherin(P<0.01,P<0.001),and upregulated the expression of E-Cadherin protein.In addition,BA inhibited the expression of β-catenin and p-GSK3β proteins.Wnt agonist LiCl could significantly antagonize the anti-colon cancer effect of BA;Wnt inhibitor XAV939 could enhance the anti-colon cancer effect of BA.In the in vivo experiment,compared with the blank group,the tumor volume of the low-dose and high-dose BA groups was significantly reduced(P<0.05,P<0.001).Immunohistochemistry results showed that compared with the blank group,the expression of Ki67 in tumor tissues of the low-dose and high-dose BA groups was significantly reduced(P<0.001).Western blot results further proved that BA inhibited the Wnt/β-catenin signaling pathway.CONCLUSION BA inhibits the viability,invasion,and migration of colon cancer HT-29 cells,induces apoptosis,and causes cell cycle arrest.Additionally,it significantly suppresses the growth of subcutaneous HT-29 cell xenografts in vivo,possibly related to the Wnt/β-catenin signaling pathway.

Objective To explore the correlation between the diameter of patients'autologous tendon and the prognosis of anterior cruciate ligament(ACL)reconstruction surgery.Methods From October 2018 to October 2023,82 patients who underwent anterior cruciate ligament reconstruction surgery at Wuxi No.904 Hospital were selected.Based on whether complications occurred after the anterior cruciate ligament reconstruction surgery,they were divided into a group with good prognosis(n=64)and a group with poor prognosis(n=18).Performing univariate analysis on the prognosis of anterior cruciate ligament reconstruction surgery.Pearson correlation analysis was used to analyze the relationship between tendon diameter and prognosis.A multifactorial Logistic regression model was applied to analyze the factors influencing the prognosis of ACL reconstruction and construct a nomogram.The diagnostic efficacy of tendon diameter on the prognosis of ACL reconstruction was analyzed using the receiver operating characteristic(ROC)curve.Results The results of the univariate analysis showed that there were statistically significant differences in gender and age between the two groups(P＜0.05).The tendon diameter and cross-sectional area in the group with a good prognosis were higher than those in the group with a poor prognosis(P＜0.05).Post-treatment,the Lysholm,Tegner,and IKDC scores in the group with a good prognosis were higher than those in the group with a poor prognosis(P＜0.05).Pearson correlation analysis showed a significant positive correlation between the patient's tendon diameter and Lysholm,Tegner,and IKDC scores(r=0.417,0.411、0.446,P＜0.05).Logistic analysis indicated that gender,age,tendon diameter,and tendon cross-sectional area are factors affecting the prognosis of ACL reconstruction(P＜0.05).The ROC curve results showed that the combined diagnostic effect of gender,age,tendon diameter,and cross-sectional area is significantly better than any single factor alone.Conclusion There is a significant correlation between the patient's autologous tendon diameter and the prognosis of ACL reconstruction surgery.An increase in autologous tendon diameter enhances knee joint function and improves patient prognosis.

Objective:This study aimed to explore the clinical value of myostatin(MST) and follistatin(FST) as biological biomarkers in evaluating sarcopenia in elderly women.Methods:This was a retrospective cross-sectional study that enrolled 350 females aged 20-89 years who underwent physical examinations in Shanghai Huadong Hospital in 2021. Demographic characteristics, muscle mass, fat mass, bone mineral density, hand grip strength, gait speed, and serum indices of MST and FST were collected.Results:The serum levels of MST did not change significantly with age. However, the serum levels of FST increased with age. In women aged≥60 years, MST was positively correlated with total lean mass and appendicular skeletal muscle index(ASMI; r=0.236, P=0.041; r=0.289, P=0.014), while FST was negatively correlated with ASMI( r=-0.265, P=0.030). In multivariate stepwise regression analysis, after adjusting for age, body mass index, hip bone mineral density, and total fat mass, only FST was independently correlated with ASMI( β=-0.238, P=0.006), while MST was not correlated with ASMI. The receiver operating characteristic curve was plotted using muscle mass reduction as the state variable and serum FST level as the test variable. The area under the curve was 0.753. And when the FST cutoff value was 17.49 ng/mL, the maximum Jordan index was 0.46, with a sensitivity of 77.3% and a specificity of 68.7%. Women aged ≥60 years were divided into three groups based on serum FST levels. Compared to the upper third of the serum FST level group, the low third of the FST level group had a significantly reduced risk of suffering from sarcopenia( OR=0.098, P =0.036). Conclusions:Serum FST lever has a better correlation with muscle mass among elderly women, making it a promising biomarker for evaluating muscle mass.

Objective: To investigate the exposure to television advertising of sugar sweetened beverages and the use of persuasive marketing techniques among children and adolescents in Beijing, so as to provide evidence for reduing childrens intake of sugar sweetened beverages. Methods: From October 19, 2020 to November 16, 2021, 32 days were randomly selected. The top four popular channels of children and adolescents aged 3-18 years were defined. Each channel was monitored from 6:00:00 to 23:59:59 for each date. A total of 2 304 h was recorded. Advertisements involving sugar sweetened beverages broadcast before, during or after the program were included. The frequency and the use of persuasive marketing techniques were analyzed. Results: A total of 1 237 advertisements for sugar sweetened beverages were included, of which 50.93% were dairy beverages, 28.38% were teabased beverages, and 19.48% were vegetable protein beverages. The average frequency of sugar sweetened beverages advertisements on every channel was (0.62±1.29)piece/h. The frequency of sugar sweetened beverages advertisements on every local channel [(1.04±1.35)piece/h] and childrens channel [(1.11±1.61)piece/h] was separately higher than every national channel [(0.48±1.24)piece/h] and general channel [(0.12±0.48)piece/h] (t=-14.05, 31.64, P<0.01). There were seasonal differences in television advertising of sugar sweetened beverages, and were more frequent during lunch and dinner times. The most frequently used persuasive marketing techniques were "images of children" (74.54%), "nutritional message" (61.76%), "product composition details" (58.61%), "nutrition claim" (57.24%), and "nutrition function claim or other function claim" (53.11%). Conclusions Children and adolescents are often exposed to television advertisement of sugar sweetened beverages on childrens channels and during meal times. There is an urgent need to formulate relevant policies to regulate the marketing of sugar sweetened beverages advertisement and reduce children and adolescents intake.

Objective To investigate the expression and clinical significance of mismatch repair (MMR) protein,human epidermal growth factor receptor (HER)2 and Ki-67 in colorectal cancer.Methods The clini-cal data of 559 patients with colorectal cancer who underwent surgical treatment in the hospital from October 2017 to May 2022 were retrospectively analyzed.The expressions of MMR protein (MLH1,MSH2,PMS2, MSH6),HER2 and Ki-67 in colorectal cancer were detected by immunohistochemistry,and the relationship between them and the clinicopathological features of colorectal cancer was analyzed.Results Among the 559 patients with colorectal cancer,43 cases (7.7％) were deficient mismatch repair (dMMR),8 cases(1.4％) were HER2 +++,and 251 cases (44.9％) were Ki-67 +++.The expression rate of dMMR in the colorec-tal cancer patients with different age,tumor location,tumor maximum diameter,gross type,histological grade,lymph node metastasis and TNM stage was significantly different (P<0.05).The expression rate of HER2 ++/+++ in the patients with different gross types of colorectal cancer was statistically significant (P<0.05).The expression rate of Ki-67 +++ in the colorectal cancer patients with different histological types,histological grades,lymph node metastasis,distant metastasis and TNM staging was statistically signifi-cant (P<0.05).There was no correlation between MMR protein and the expression of HER2 and Ki-67 (P>0.05).Conclusion The expression of MMR proteins in colorectal cancer is closely related to its clinicopatho-logical features.

Objective To explore efficacy and safety of integrated traditional Chinese and western medicine in the treatment of intrahepatic cholestasis of pregnancy(ICP).Methods Clinical data of 151 pregnant women with ICP received treatment at Affiliated Hospital of Hangzhou Normal University from January 2020 to June 2022 were retrospectively analyzed.According to the differences in treatment plans,all patients were divided into control group(100 cases)and observation group(51 cases).Control group received basic therapy,including ursodeoxycholic acid capsules and adenosylmethionine for injection.Observation group was given Yinchenhao decoction with modified for one week on the basis of treatment in control group.Serum indicators of laboratories,itching status,maternal and infant outcomes,and safety between two groups were compared.Results After treatment,serum total bilirubin,alanine transaminase,aspartate transaminase,total bile acid levels,itching score in two groups were significantly lower than those before treatment,and observation group was lower than control group(P<0.05).Itch relief rate in observation group was significantly higher than that in control group,and itch control time in observation group was significantly shorter than that in control group(P<0.05).Incidence rate of cesarean section,amniotic fluid turbidity and postpartum hemorrhage in observation group were significantly lower than those in control group,and gestational age at termination of pregnancy in observation group was significantly higher than that in control group(P<0.05).After treatment,ratio of peak systolic and end diastolic flow velocity of the umbilical artery in two groups were significantly lower than that before treatment,and observation group was lower than control group(P<0.05).No drug allergic reactions occurred during treatment of two groups,and there were no serious complications such as placental abruption or stillbirth.Conclusion Compared to a single western medicine therapy,combination of Chinese and western medicine has a better effect on treating ICP.It can greatly improve levels of serum bile acids,bilirubin and liver enzymes,reduce itching,and improve maternal and infant outcomes.It has minimal side effects,and has good safety.

Objective To explore the correlation between the diameter of patients'autologous tendon and the prognosis of anterior cruciate ligament(ACL)reconstruction surgery.Methods From October 2018 to October 2023,82 patients who underwent anterior cruciate ligament reconstruction surgery at Wuxi No.904 Hospital were selected.Based on whether complications occurred after the anterior cruciate ligament reconstruction surgery,they were divided into a group with good prognosis(n=64)and a group with poor prognosis(n=18).Performing univariate analysis on the prognosis of anterior cruciate ligament reconstruction surgery.Pearson correlation analysis was used to analyze the relationship between tendon diameter and prognosis.A multifactorial Logistic regression model was applied to analyze the factors influencing the prognosis of ACL reconstruction and construct a nomogram.The diagnostic efficacy of tendon diameter on the prognosis of ACL reconstruction was analyzed using the receiver operating characteristic(ROC)curve.Results The results of the univariate analysis showed that there were statistically significant differences in gender and age between the two groups(P＜0.05).The tendon diameter and cross-sectional area in the group with a good prognosis were higher than those in the group with a poor prognosis(P＜0.05).Post-treatment,the Lysholm,Tegner,and IKDC scores in the group with a good prognosis were higher than those in the group with a poor prognosis(P＜0.05).Pearson correlation analysis showed a significant positive correlation between the patient's tendon diameter and Lysholm,Tegner,and IKDC scores(r=0.417,0.411、0.446,P＜0.05).Logistic analysis indicated that gender,age,tendon diameter,and tendon cross-sectional area are factors affecting the prognosis of ACL reconstruction(P＜0.05).The ROC curve results showed that the combined diagnostic effect of gender,age,tendon diameter,and cross-sectional area is significantly better than any single factor alone.Conclusion There is a significant correlation between the patient's autologous tendon diameter and the prognosis of ACL reconstruction surgery.An increase in autologous tendon diameter enhances knee joint function and improves patient prognosis.