Objective To understand the genotyping of human immunodeficiency virus (HIV) co-infected hepatitis C virus (HCV) patients in Yunnan Province, and to analyze the differences in viral load, biochemical indicators, and blood routine indicators among different genotypes, in order to provide a laboratory basis for the diagnosis and clinical treatment of HIV/HCV co-infected patients. Methods From November 2022 to June 2023, the serum samples and basic information of patients diagnosed with HIV/HCV co-infection were collected in the antiviral outpatient clinic of Yunnan Provincial Hospital of Infectious Diseases. The HCV viral load was detected by one-step qRT-PCR amplification, the positive samples were sequenced, and genotyping was determined based on NS5 gene sequence. The differences in biochemical and blood routine indexes between HIV patients co-infected with different HCV genotypes and low/high viral loads were analyzed. Results A total of 126 HIV/HCV co-infected patients were collected, including 20 HCV genotype 1 (15.9%), 91 HCV genotype 3 (72.2%), and 15 HCV genotype 6 (11.9%). The maximum and minimum viral load of the three HCV genotypes were as follows: HCV type 1 (1.0×108, 4.8×104 IU/mL), HCV type 3 (2.2×108, 2.9×102 IU/mL), and HCV type 6 (8.1×107, 6.8×104 IU/mL). The results showed that there was no significant difference between HIV co-infection with different genotypes of HCV and three HIV treatment schemes, including nucleoside reverse transcriptase inhibitors+integrase strand transfer inhibitors (NRTIs+INSTIs), nucleoside reverse transcriptase inhibitors+non-nucleoside reverse transcriptase inhibitors (NRTIs+NNRTIs) and nucleoside reverse transcriptase inhibitors+protease inhibitor (NRTIs+PLs), and the viral load of patients (P>0.05). The analysis of biochemical indexes such as total bilirubin (TBIL), direct bilirubin (DBIL), alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatinine (CREA), and blood routine indexes such as white blood cell (WBC), red blood cell (RBC), hemoglobin (HGB), platelet (PLT), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) among different HCV genotypes and low/high viral loads showed that there was no significant difference in biochemical indexes and blood routine indexes between low/high viral loads of HIV co-infected HCV patients (P>0.05); however, the biochemical indicators TBIL, IBIL and MCHC were significantly different statistically between patients with genotype 3 HCV infection and those with genotype 1 HCV infection (P<0.05), while other biochemical and blood routine indexes were not statistically different among different HCV genotypes (P>0.05). Conclusions There are six subtypes of HCV co-infection in HIV patients in Kunming, Yunnan Province, including three genes of genotype 1, 3, and 6. Among them, genotype 3 HCV is the main prevalent genetic virus among HIV co-infected populations. The TBIL, IBIL and MCHC values of HIV patients co-infected with HCV type 3 are different from those infected with HCV type 1.

AIM:This study examined the inhibitory effect of peiminine on the human colonic adenocarcino-ma cell line SW480 and explored the underlying mechanisms.METHODS:SW480 and human normal colonic epithelial CCD-841CoN cells were treated with different concentrations of peiminine and subjected to the CCK-8 assay to select the optimal treatment time and concentration of the compound.SW480 cell migration and invasion were evaluated by the wound-healing and Transwell assays.Cell cycle progression was analyzed by flow cytometry.The expression levels of cell cycle-related proteins were examined by Western blot.SW480 xenograft tumor model was established in nude mice to ex-amine the effect of peiminine on tumor growth and the expression of cell cycle-related proteins in vivo.RESULTS:Peimi-nine(110 mg/L)significantly inhibited the proliferation of SW480 cells compared with the control group(P<0.01),caused cell cycle arrest at G1 phase,and significantly downregulated the expression of cyclin dependent kinase 2(CDK2),CDK4,CDK6,cyclin D1,p-Rb/Rb,E2F1,E2F3,and E2F4(P<0.05).Peiminine inhibited SW480 xenograft tumor growth,prolonged the survival of model mice,and affected the expression of CDK2,CDK4,CDK6,and cyclin D1 in tu-mor tissues.CONCLUSION:Peiminine promotes G1 phase arrest by down-regulating the expression of CDK2,CDK4,CDK6,and cyclin D1,thereby inhibiting the proliferation of SW480 cells.

AIM:To explore the effect and mechanism of action of the aqueous extract of Fritillaria ussuriensis(FU-AE)against nonalcoholic fatty liver disease(NAFLD).METHODS:The association between Fritillaria ussuriensis Maxir.(FU)and NAFLD was analyzed by network pharmacology.A mouse model of NAFLD was induced in mice by high fat diet(HFD)+10%fructose drinking water,and three doses of Fritillaria ussuriensis aqueous extract were given to the mice for intervention.Colorimetric assay was used for detection of aspartate aminotransferase(AST),alanine aminotrans-ferase(ALT),triglyceride(TG),total cholesterol(TC),high-density lipoprotein cholesterol(HDL-C),and low-density lipoprotein cholesterol(LDL-C)levels in the serum of experimental mice.Hematoxylin and eosin staining was used to as-sess the pathological and histological changes in the liver of mice and to clarify the anti-NAFLD effect of aqueous extracts of Fritillaria ussuriensis.Liver tissue proteins were extracted,and expression of proteins related to the AMP-activated pro-tein kinase(AMPK)/acetyl-CoA carboxylase(ACC)pathway was detected by Western blot to clarify the mechanism of an-ti-NAFLD action of Fritillaria ussuriensis.The microbial composition of cecum contents was explored using 16S rRNA se-quencing to reveal the modulatory effect of the aqueous extract of Fritillaria ussuriensis on the structure of intestinal flora in mice with nonalcoholic fatty liver disease.RESULTS:Aqueous extract of Fritillaria ussuriensis(high dose)ameliorated exogenous adipocyte infiltration in the liver of mice with NAFLD(P<0.05).AST,ALT,TG,TC and LDL-C levels were significantly decreased(P<0.05)and HDL-C levels were significantly increased(P<0.05)in the high-dose group.Aque-ous extract of Fritillaria ussuriensis(high dose)significantly increased expression of phosphorylated AMPKα,AMPKα,and phosphorylated ACC in the livers of the model mice(P<0.05),significantly reduced expression of ACC(P<0.05),and significantly increased the relative abundance of the potentially beneficial bacteria Faecalibaculum rodentium,Lacto-bacillus johnsonii,Akkermansia muciniphila(P<0.05).CONCLUSION:Aqueous extract of Fritillaria ussuriensis may ameliorate NAFLD in mice by activating the AMPK/ACC pathway and modulating the structure of intestinal flora.

Objective:To investigate the association of metabolic syndrome(MS) with cardiovascular disease(CVD) mortality and all-cause mortality in peritoneal dialysis patients.Methods:A retrospective analysis was performed on patients who underwent peritoneal dialysis from January 1, 2013 to July 31, 2021 in the Shaoxing People′s Hospital. Patients were divided into MS group and non-MS group. The differences in baseline biochemical variables, comorbidities, and clinical outcomes between the two groups were compared. Kaplan-Meier method was used to obtain survival curves, the Cox regression model was used to evaluate the influence of MS for survival rates, and the inverse probability of treatment weighting(IPTW) was used to eliminate influence of the confounders in the groups.Results:A total of 494 peritoneal dialysis patients were enrolled in this study, which were divided into MS group( n=266) and non-MS group( n=228). The total median follow-up time was(31±22) months. At baseline, the standard mean difference( SMD) in smoking history, drinking history, CVD history, prevalence of chronic glomerulonephritis, left ventricular ejection fraction, B-type natriuretic peptides, hemoglobin, blood calcium, hypersensitive C-reactive-protein, intact parathyroid hormone, ultrafiltration and 4 h dialysate/plasma creatinine in the two groups were greater than 0.1. Their SMD decreased to under 0.1 after IPTW, showing a good balance between the two groups. The analysis of the survival curve of Kaplan Meier showed that the cumulative survival rate and cumulative CVD survival rate in MS group were significantly lower than those in non-MS group before and after IPTW( P<0.05). After IPTW was used to eliminate the effect of confounders, multivariate Cox regression analysis still displayed that MS was an independent risk factor for all-cause mortality( HR=1.824, 95% CI 1.121-2.968, P=0.015) and CVD mortality( HR=2.470, 95% CI 1.324-4.609, P=0.004)in peritoneal dialysis patients. Conclusion:The prevalence of metabolic syndrome is high in peritoneal dialysis patients. MS is an independent risk factor for all-cause mortality and CVD mortality in peritoneal dialysis patients.

Objective To investigate the relationship between (serum neutrophil gelatinase-associated lipocalin,sNGAL) and cardiovascular events in patients with chronic kidney disease(CKD).Methods 300 patients with CKD were divided into two groups according to the level of sNGAL:high sNGAL group (n=158) and low sNGAL group (n=142).The incidence of cardiovascular events and cumulative survival rate were analyzed by ROC curve,and the correlation between sNGAL and cardiovascular risk factors,cardiovascular events in patients with chronic renal disease was analyzed.Influencing factors of cardiovascular events in CKD patients was analyzed.Results There were significant differences in the data about BMI,diabetes proportion,CKD staging,eGFR,hsCRP,24h proteinuria,HDL,iPTH,phosphate and blood calcium between the two groups (P＜0.05).The 3-year cumulative survival rate of high sNGAL group(77.2％) was significantly lower than that of low sNGAL group(96.5％),and the 3-year incidence of cardiovascular events (37.9％) was significantly higher than that of low sNGAL group (9.8％) (P＜ 0.05).AUC in diagnosing cardiovascular events in high sNGAL group (0.746) was significantly higher than that in eGFR(0.636),age (0.504),serum calcium (0.545),HDL(0.594) and LDL (0.508,all P＜0.05).There was a significant correlation between sNGAL and eGFR,HDL,BMI,hs-CRP,iPTH and phosphate (P＜ 0.05).Both univariate and multivariate fact ors COX showed that sNGAL was a risk factor of cardiovascular events in patients with CKD (P＜0.05),((HR=1.976 and 1.588,95％ CI=1.443-2.724 and 1.144-2.143,respectively,P=0.O00 and 0.000)).Conclusions The incidence of cardiovascular events in patients with CKD with high sNGAL is significantly increased.sNGAL is an independent factor of cardiovascular events in patients with chronic renal disease.

Objective: To investigate the diagnosis and treatment for enterocutaneous fistula (ECF) in China, and to explore the prognostic factors of ECF. Methods: A multi-center cross-sectional study was conducted based on the Registration System of Chinese Gastrointestinal Fistula and Intra-Abdominal Infections to collect the clinical data of ECF patients from 54 medical centers in 22 provinces/municipalities from January 1, 2018 to December 31, 2018. The clinical data included patient gender, age, length of hospital stay, intensive care unit (ICU) admission, underlying diseases, primary diseases, direct causes of ECF, location and type of ECF, complications, treatment and outcomes. All medical records were carefully filled in by the attending physicians, and then re-examined by more than two specialists. The diagnosis of ECF was based on the clinical manifestations, laboratory/imaging findings and intraoperative exploration. Results: A total of 1521 patients with ECF were enrolled, including 1099 males and 422 females, with a median age of 55 years. The top three primary diseases of ECF were malignant tumors in 626 cases (41.2%, including 540 gastrointestinal tumors, accounting for 86.3% of malignant tumors), gastrointestinal ulcers and perforations in 202 cases (13.3%), and trauma in 157 cases (10.3%). The direct causes of ECF were mainly surgical operation in 1194 cases (78.5%), followed by trauma in 156 (10.3%), spontaneous fistula due to Crohn′s disease in 92 (6.0%), radiation intestinal injury in 41 (2.7%), severe pancreatitis in 20 (1.3%), endoscopic treatment in 13 (0.9%) and 5 cases (0.3%) of unknown reasons. All the patients were divided into three groups: 1350 cases (88.7%) with simple ECF, 150 (9.9%) with multiple ECF, and 21 (1.4%) with combined internal fistula. Among the patients with simple ECF, 438 cases (28.8%) were jejuno-ileal fistula, 313 (20.6%) colon fistula, 170 (11.2%) rectal fistula, 111 (7.3%) duodenal fistula, 76 (5.0%) ileocecal fistula, 65 (4.3%) ileocolic anastomotic fistula, 55 (3.6%) duodenal stump fistula, 36 (2.4%) gastrointestinal anastomotic fistula, 36 (2.4%) esophagogastric/esophagojejunal anastomotic fistula, 29 (1.9%) gastric fistula and 21 (1.4%) cholangiopancreatiointestinal. Among all the simple ECF patients, 991 were tubular fistula and 359 were labial fistula. A total of 1146 patients finished the treatment, of whom 1061 (92.6%) were healed (586 by surgery and 475 self-healing) and 85 (7.4%) died. A total of 1043 patients (91.0%) received nutritional support therapy, and 77 (6.7%) received fistuloclysis. Infectious source control procedures were applied to 1042 patients, including 711 (62.0%) with active lavage and drainage and 331 (28.9%) with passive drainage. Among them, 841 patients (73.4%) underwent minimally invasive procedures of infectious source control (replacement of drainage tube through sinus tract, puncture drainage, etc.), 201 (17.5%) underwent laparotomy drainage, while 104 (9.1%) did not undergo any drainage measures. A total of 610 patients (53.2%) received definitive operation, 24 patients died within postoperative 30-day with mortality of 3.9% (24/610), 69 (11.3%) developed surgical site infection (SSI), and 24 (3.9%) had a relapse of fistula. The highest cure rate was achieved in ileocecal fistula (100%), followed by rectal fistula (96.2%, 128/133) and duodenal stump fistula (95.7%,44/46). The highest mortality was found in combined internal fistula (3/12) and no death in ileocecal fistula. Univariate prognostic analysis showed that primary diseases as Crohn′s disease (χ²=6.570, P=0.010) and appendicitis/appendiceal abscess (P=0.012), intestinal fistula combining with internal fistula (χ²=5.460, P=0.019), multiple ECF (χ²=7.135, P=0.008), esophagogastric / esophagojejunal anastomotic fistula (χ²=9.501, P=0.002), ECF at ileocecal junction (P=0.012), non-drainage/passive drainage before the diagnosis of intestinal fistula (χ²=9.688, P=0.008), non-drainage/passive drainage after the diagnosis of intestinal fistula (χ²=9.711, P=0.008), complicating with multiple organ dysfunction syndrome (MODS) (χ²=179.699, P<0.001), sepsis (χ²=211.851, P<0.001), hemorrhage (χ²=85.300, P<0.001), pulmonary infection (χ²=60.096, P<0.001), catheter-associated infection (χ²=10.617, P=0.001) and malnutrition (χ²=21.199, P<0.001) were associated with mortality. Multivariate prognostic analysis cofirmed that sepsis (OR=7.103, 95%CI:3.694-13.657, P<0.001), complicating with MODS (OR=5.018, 95%CI:2.170-11.604, P<0.001), and hemorrhage (OR=4.703, 95%CI: 2.300-9.618, P<0.001) were independent risk factors of the death for ECF patients. Meanwhile, active lavage and drainage after the definite ECF diagnosis was the protective factor (OR=0.223, 95%CI: 0.067-0.745, P=0.015). Conclusions The overall mortality of ECF is still high. Surgical operation is the most common cause of ECF. Complications e.g. sepsis, MODS, hemorrhage, and catheter-associated infection, are the main causes of death. Active lavage and drainage is important to improve the prognosis of ECF.

Objective To evaluate the role of spinal nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling pathway in hydrogen-induced reduction of inflammatory pain in rats.Methods Sixty-four SPF healthy adult male Sprague-Dawley rats,weighing 200-250 g,were divided into 4 groups (n =16 each) using a random number table:control group (group C),inflammatory pain group (group IP),inflammatory pain plus hydrogen-rich saline group (group IP+H2) and inflammatory pain plus hydrogen-rich saline plus Nrf2 inhibitor all-trans retinoic acid (ATRA) group (group IP+H2+ATRA).Chronic inflammatory pain was induced by injecting complete Freund's adjuvant (CFA) 100 μl into the plantar surface of the left hind paw in IP group and IP+H2 group.The equal volume of normal saline was given instead in group C.Hydrogen-rich saline 5 ml/kg was injected intraperitoneally once a day for 7consecutive days starting from 1 day after injecting CFA in group IP+H2 and group IP+H2+ATRA,and the equal volume of normal saline was given instead in the other groups.ATRA 7 mg/kg was injected intraperitoneally once a day for 2 consecutive days starting from 2 days before injecting CFA.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 1 day before establishing the model (T0) and 1,3 and 7 days after establishing the model (T1-3).Six rats were sacrificed after the last measurement of pain threshold on day 7 after establishing the model,and the L4-6 lumbar segments of the spinal cord were removed for determination of the expression of Nrf2,HO-1 and glial fibrillary acidic protein (GFAP) by Western blot.Results Compared with group C,the MWT was significantly decreased and the TWL was shortened at T1-3,and the expression of Nrf2,HO-1 and GFAP was up-regulated in IP and IP+H2 groups (P＜0.05).Compared with group IP,the MWT was significantly increased and the TWL was prolonged at T1-3,the expression of Nrf2 and HO-1 was up-regulated,and the expression of GFAP was down-regulated in group IP+H2 (P＜0.05),and no significant change was found in the parameters mentioned above in group IP+H2+ATRA (P＞0.05).Compared with group IP+H2,the MWT was significantly decreased and the TWL was shortened at T1-3,the expression of Nrf2 and HO-1 was down-regulated,and the expression of GFAP was up-regulated in group IP+H2+ATRA (P＜0.05).Conclusion Activation of spinal Nrf2/HO-1 signaling pathway is involved in hydrogen-induced reduction of infflammatory pain in rats.

This study was aimed to analyze the application of non-pharmacotherapy in treating cervical radiculopathy (CR) in real-world,and to provide clinical reference for CR non-pharmacotherapy.The clinical real-world data of CR was extracted by using information sharing system of traditional Chinese medicine (TCM) and clinical research.Six hundred and twenty-eight inpatients and outpatients with CR were enrolled from December 2012 to July 2014 in the information system database of Wangjing Hospital.Basic characteristics of the non-pharmacotherapy groups were analyzed by statistical description method.The node degree and mutual information value were recorded for non-pharmacotherapy application of all patients by using liquorice software.Complex network diagrams were generated.The results showed that 47％ of CR patients received non-pharmacotherapy (294/628),including 67 males and 227 females.The average age of patients was 49 years old,and the prevalence of the disease was the highest from 45 to 65 years old.In all patients,the usage of manipulation and cervical traction was higher,and the combination of manipulation and acupuncture was the most.Within outpatients,the proportion of cervical traction was higher,and the combination of manipulation and acupuncture was the most frequently.Within inpatients,the proportion of manipulation and cervical traction was higher,and the combination of comprehensive physical therapy and exercise therapy was the most frequently.It was concluded that non-pharmacotherapy has been commonly used in clinical treatment of CR.Cervical traction and manipulation was the widest applications.The combination treatment was in wide application.Future studies should increase the sample size of CR patients from different regions,and enhance gradually the level of evidence of clinical research for non-pharmacotherapy treating CR.

Objective:To explore the inhibitory effects of seabuckthorn polysaccharide on hepatic oxidative stress in a mice model of acute liver injury induced by intraperitoneal injection of LPS and D -GalN and detect the expression on hepatic BCL-2/Bax and PPAR-γ.Methods: C57BL/6 male mice were randomly divided into six groups:control group ( CTRL), model group ( L/G), dexamethasone positive control group ( DXM ) , low ( SPL ) , medium ( SPM ) and high dose group ( SPH ) of seabuckthorn polysaccharide.Mice in the SPL,SPM and SPH group were gavaged with 50,100 and 200 mg/kg seabuckthorn polysaccharide for 14 days respectively.Acute liver injury model were established by intraperitoneal injection of LPS (10 μg/kg) and D-GalN (700 mg/kg) .Serum and liver samples were collected 4 h after model establishment .Serum levels of ALT and AST and the content of MDA were de-tected.Hepatic expression of SOD 2 BCL-2 and Bax was determined by Western blot and the expression of PPAR-γwas detected by im-munohistochemistry .Results:ALT and AST levels significantly increased in the model group and decreased dose-dependently after pre-treatment with seabuckthorn polysaccharide .The level of MDA in the model group increased significantly as compared with the control group and decreased in seabuckthorn polysaccharide groups ,while the level of SOD 2 decreased in the model group and recovered in sea-buckthorn polysaccharide groups .The expression of Bax decreased after pretreatment with seabuckthorn polysaccharide .There was no obvious effect on BCL-2 expression after sea buckthorn polysaccharide supplementation .The expression of PPAR-γreduced in the sea-buckthorn polysaccharide group as compared with the model group .Conclusion:Seabuckthorn polysaccharide protects against LPS /D-GalN-induced liver injury.The effect is associated with an upregulation of SOD 2 and downregulation of Bax .

Objective:To establish cell lines stably expressing Rab5a and its the inactive mutant Rab5aN133I,analyze the effect of Rab5a on the expression of cytokines in LPS-stimulated RAW264.7 cells .Methods:RAW264.7 cells were transfected with Rab5a and its inactive mutant vector Rab5a N133I separately,and then screened by G418.Rab5a stable expressing cell lines were identified by Real time-PCR.The growth of the stable cell lines was analyzed by MTT assay.After the stable cell lines were stimulated by LPS for different time periods,the expression of iNOS,TNF-αand IL-6 was detected.Results:Rab5a and Rab5aN133I transfection resulted in elevated Rab5a mRNA expression compared with the control cells ( P<0.05 ).Rab5a overexpression enhanced the proliferation of RAW264.7 cells.However,the proliferation of Rab5aN133I cells was significantly slower than the control cells ( P<0.05).Overexpression of Rab5a promoted LPS-induced production of iNOS,TNF-αand IL-6 in RAW264.7 cells (P<0.01). Conversely,overexpression of Rab5aN133I abolished the stimulating effects of Rab5a.Conclusion: Rab5a promoted LPS-induced expression of iNOS,TNF-αand IL-6 in RAW264.7 macrophages in a GTP-binding ability-dependent manner.