To verify whether chaperones can promote the soluble expression of PlpE in Escherichia coli and whether the expressed protein is active,prokaryotic expression and Western blot detection were performed.The results showed that:The PlpE prokaryotic expression vector pET-32a(+)-plpE was expressed as inclusion body,and the expression form was not changed by changing the concentration of inducer,induction time and temperature.The companion proteins pG-KJE8,pGro7,pKJE7 and pG-Tf2 were co-expressed with pET-32a(+)-plpE in Eschierichia coli expres-sion system,respectively.When the final concentration of IPTG of 0.5 mmol/L,L-arabinose of 0.5 g/L or tetracycline of 5.0 μg/L were added as inducers and induced at 37 ℃ for 8 h,the results showed that the molecular companion pGro7 could change the expression of rp-PlpE from inclu-sion body to soluble expression.pG-KJE8,pKJE7 and pG-Tf2 had no effect on the expression of rp-PlpE.The soluble rp-PlpE can react specifically with the positive serum of Mannheimia haemolyti-ca.Therefore,the study showed that the co-expression of the chaperone protein pGro7 can make the rp-PlpE protein express in a soluble form,and the purified protein exhibits reactogenicity.These findings lay the foundation for the establishment of a subunit vaccine and serological diagno-sis methods for Mannheimia haemolytica.

This article explores the compatibility of traditional Chinese medicine for external treatment of ulcers from the perspec-tive of"the principle of internal and external treatment"in Rhymed Discourse on External Remedies.First,it discusses the clinical un-derstanding of chronic skin ulcers,and then it explains the connotation of"the principle of internal and external treatment",which has the characteristics of theoretical homology,syndrome differentiation and treatment,and focusing on the property and taste of the medi-cine first.Finally,based on"the principle of internal and external treatment",it discusses the compatibility ideas of Chinese medicine for external treatment of ulcers,and specifically analyzes the compatibility ideas of property and taste and monarch,minister,assistant and envoy,so as to provide some reference for the external treatment and compatibility of Chinese medicine for chronic skin ulcers,and also hopes to provide some directions for the research and development of external traditional Chinese patent medicines and simple prep-arations.

This article explores the compatibility of traditional Chinese medicine for external treatment of ulcers from the perspec-tive of"the principle of internal and external treatment"in Rhymed Discourse on External Remedies.First,it discusses the clinical un-derstanding of chronic skin ulcers,and then it explains the connotation of"the principle of internal and external treatment",which has the characteristics of theoretical homology,syndrome differentiation and treatment,and focusing on the property and taste of the medi-cine first.Finally,based on"the principle of internal and external treatment",it discusses the compatibility ideas of Chinese medicine for external treatment of ulcers,and specifically analyzes the compatibility ideas of property and taste and monarch,minister,assistant and envoy,so as to provide some reference for the external treatment and compatibility of Chinese medicine for chronic skin ulcers,and also hopes to provide some directions for the research and development of external traditional Chinese patent medicines and simple prep-arations.

To verify whether chaperones can promote the soluble expression of PlpE in Escherichia coli and whether the expressed protein is active,prokaryotic expression and Western blot detection were performed.The results showed that:The PlpE prokaryotic expression vector pET-32a(+)-plpE was expressed as inclusion body,and the expression form was not changed by changing the concentration of inducer,induction time and temperature.The companion proteins pG-KJE8,pGro7,pKJE7 and pG-Tf2 were co-expressed with pET-32a(+)-plpE in Eschierichia coli expres-sion system,respectively.When the final concentration of IPTG of 0.5 mmol/L,L-arabinose of 0.5 g/L or tetracycline of 5.0 μg/L were added as inducers and induced at 37 ℃ for 8 h,the results showed that the molecular companion pGro7 could change the expression of rp-PlpE from inclu-sion body to soluble expression.pG-KJE8,pKJE7 and pG-Tf2 had no effect on the expression of rp-PlpE.The soluble rp-PlpE can react specifically with the positive serum of Mannheimia haemolyti-ca.Therefore,the study showed that the co-expression of the chaperone protein pGro7 can make the rp-PlpE protein express in a soluble form,and the purified protein exhibits reactogenicity.These findings lay the foundation for the establishment of a subunit vaccine and serological diagno-sis methods for Mannheimia haemolytica.

Objective:To investigate the effect of laparoscopic splenectomy and azygoportal disconnection (LSD) on liver synthesis and development of liver cirrhosis.Methods:This is a prospective case series study.The clinical data of liver cirrhotic patients with portal hypertension who received LSD at the Department of Hepatobiliary Surgery of Northern Jiangsu People′s Hospital Affiliated to Yangzhou University from September 2014 to January 2016 were included. This study analyzed the diameter of the portal vein, the velocity of portal blood flow, the routine blood parameters, the liver function, the synthetic proteins of liver (antithrombin Ⅲ (AT-Ⅲ), protein S, protein C), and the serum content of liver fibrotic markers(collagen type Ⅳ, procollagen type Ⅲ, laminin, hyaluronidase). Repeated measures ANOVA was used for comparison between multiple groups, and least significance difference was used for post-hoc multiple comparison.Results:A total of 106 patients were included in the study, including 70 males and 36 females, aged (51.8±9.8) years(range: 28 to 75 years).Compared with the preoperative results, the diameter of portal vein and the velocity of portal vein decreased after surgery ( F=14.03, 12.15, respectively, both P<0.01). Compared with the preoperative results, the total bilirubin, albumin, prothrombin time, international normalized ratio, Child-Pugh score and classification were improved ( F=17.96, 56.01, 66.63, 35.83, 33.49, and 27.50, respectively, all P<0.01), and the AT-Ⅲ, protein S, protein C,collagen type Ⅳ, procollagen type Ⅲ, laminin and hyaluronidase levels were also improved ( F=47.87, 36.26, 18.02, 2.79, 14.58, 44.35, and 14.38, respectively, all P<0.01). Compared with the preoperative period, the diameter of portal vein was reduced from the first week to the 24 th month after surgery ( t=5.45 to 9.39, all P<0.01). Compared with the preoperative period, the velocity of portal vein blood from the first week after surgery to the 24 th month after surgery was decreased ( t=4.02 to 8.43, all P<0.01). Compared with the preoperative period, routine blood parameters (white blood count, hemoglobin, platelet count), liver function (total bilirubin, albumin, prothrombin time, international normalized ratio, Child-Pugh score), liver synthetic protein (AT-Ⅲ, protein S, protein C) and liver fibrotic markers (collagen type Ⅳ, procollagen type Ⅲ, laminin, hyaluronidase) were improved to varying degrees at the 24th month after surgery ( t=-20.46 to 11.93, all P<0.01). Conclusion:Preliminary findings show that LSD can reduce portal vein pressure, restore blood cell number, and improve liver synthesis function and the degree of liver fibrosis in patients with portal hypertension in liver cirrhosis.

Objective:To investigate the effect of laparoscopic splenectomy and azygoportal disconnection (LSD) on liver synthesis and development of liver cirrhosis.Methods:This is a prospective case series study.The clinical data of liver cirrhotic patients with portal hypertension who received LSD at the Department of Hepatobiliary Surgery of Northern Jiangsu People′s Hospital Affiliated to Yangzhou University from September 2014 to January 2016 were included. This study analyzed the diameter of the portal vein, the velocity of portal blood flow, the routine blood parameters, the liver function, the synthetic proteins of liver (antithrombin Ⅲ (AT-Ⅲ), protein S, protein C), and the serum content of liver fibrotic markers(collagen type Ⅳ, procollagen type Ⅲ, laminin, hyaluronidase). Repeated measures ANOVA was used for comparison between multiple groups, and least significance difference was used for post-hoc multiple comparison.Results:A total of 106 patients were included in the study, including 70 males and 36 females, aged (51.8±9.8) years(range: 28 to 75 years).Compared with the preoperative results, the diameter of portal vein and the velocity of portal vein decreased after surgery ( F=14.03, 12.15, respectively, both P<0.01). Compared with the preoperative results, the total bilirubin, albumin, prothrombin time, international normalized ratio, Child-Pugh score and classification were improved ( F=17.96, 56.01, 66.63, 35.83, 33.49, and 27.50, respectively, all P<0.01), and the AT-Ⅲ, protein S, protein C,collagen type Ⅳ, procollagen type Ⅲ, laminin and hyaluronidase levels were also improved ( F=47.87, 36.26, 18.02, 2.79, 14.58, 44.35, and 14.38, respectively, all P<0.01). Compared with the preoperative period, the diameter of portal vein was reduced from the first week to the 24 th month after surgery ( t=5.45 to 9.39, all P<0.01). Compared with the preoperative period, the velocity of portal vein blood from the first week after surgery to the 24 th month after surgery was decreased ( t=4.02 to 8.43, all P<0.01). Compared with the preoperative period, routine blood parameters (white blood count, hemoglobin, platelet count), liver function (total bilirubin, albumin, prothrombin time, international normalized ratio, Child-Pugh score), liver synthetic protein (AT-Ⅲ, protein S, protein C) and liver fibrotic markers (collagen type Ⅳ, procollagen type Ⅲ, laminin, hyaluronidase) were improved to varying degrees at the 24th month after surgery ( t=-20.46 to 11.93, all P<0.01). Conclusion:Preliminary findings show that LSD can reduce portal vein pressure, restore blood cell number, and improve liver synthesis function and the degree of liver fibrosis in patients with portal hypertension in liver cirrhosis.

Environmental toxicants have been linked to aging and age-related diseases. The emerging evidence has shown that the enhancement of detoxification gene expression is a common transcriptome marker of long-lived mice, Drosophila melanogaster, and Caenorhabditis elegans. Meanwhile, the resistance to toxicants was increased in long-lived animals. Here, we show that farnesoid X receptor (FXR) agonist obeticholic acid (OCA), a marketed drug for the treatment of cholestasis, may extend the lifespan and healthspan both in C. elegans and chemical-induced early senescent mice. Furthermore, OCA increased the resistance of worms to toxicants and activated the expression of detoxification genes in both mice and C. elegans. The longevity effects of OCA were attenuated in Fxr ^-/- mice and Fxr homologous nhr-8 and daf-12 mutant C. elegans. In addition, metabolome analysis revealed that OCA increased the endogenous agonist levels of the pregnane X receptor (PXR), a major nuclear receptor for detoxification regulation, in the liver of mice. Together, our findings suggest that OCA has the potential to lengthen lifespan and healthspan by activating nuclear receptor-mediated detoxification functions, thus, targeting FXR may offer to promote longevity.

Among tumor microenvironment (TME), the entire metabolic characteristics of tumor-resident cells are reprogrammed to benefit the expansion of tumor cells, which count on glutamine in large part to fuel the tricarboxylic acid cycle for energy generation and anabolic metabolism support. Endothelial cells that are abducted by tumor cells to form a pathological tumor vascular network for constructing the hypoxic immunosuppressive TME, also rely on glutaminolysis as the "engine" of angiogenesis. Additionally, the glutamine metabolic preference benefits the polarization of TAMs towards pro-tumoral M2 phenotype as well. Herein, we developed a type of siRNA micelleplexes (MH@siGLS1) to reverse immunosuppressive TME by targeting glutaminolysis within tumor-resident cells for tumor vasculature normalization- and TAMs repolarization-enhanced photo-immunotherapy. Tumor cell starvation and antioxidant system destruction achieved by MH@siGLS1-mediated glutaminolysis inhibition could promote photodynamic therapy efficacy, which was available to trigger immunogenic cell death for adaptive antitumor immune responses. Meanwhile, glutaminolysis inhibition of tumor endothelial cells and TAMs could realize tumor vascular normalization and TAMs repolarization for antitumor immunity amplification. This study provides a unique perspective on cancer treatments by focusing on the interrelations of metabolic characteristics and the biofunctions of various cell types within TME.

Objective To study the changes in peripheral blood T lymphocytes in patients with ankylosing spondylitis and their correlation with the disease's severity and prognosis.Methods We selected 120 patients with ankylosing spondylitis treated between January 2020 and March 2023 as the research group and 120 healthy people who had medical examinations in the same period as the health group.We detected the changes in CD4+,CD8+and CD4+/CD8+values of peripheral blood T lymphocyte subsets with flow cytometry,compared the differences in T lymphocyte subsets between the two groups,and analyzed the correlation with the disease severity of the patients.All the 120 patients with ankylosing spondylitis were followed up for 6 months after treatment to assess their prognosis.General information and T lymphocyte sub-groups CD4+,CD8+level,CD4+/CD8+value changes were compared among patients with different prognosis.We analyzed the value of T lymphocyte sub-groups in predicting the prognosis of patients with ankylosing spondylitis.Results In the research group CD4+and CD4+/CD8+were lower but CD8 1 was higher than those in the healthy group(P＜0.05).CD4+and CD4+/CD8 were lower but CD8+was higher in patients with advanced ankylosing spondylitis than in early and mid-term patients(P＜0.05).The ROC curve analysis showed that the AUC of CD4+,CD8+,and CD4+/CD8+combined diagnosis of ankylosing spondylitis patients was 0.878,with higher diagnostic sensitivity than that of the single diagnosis(P＜0.05).In the poor prognosis group,CD8+was higher than that in the excellent prognosis group,but CD4+and CD4+/CD8 value were lower than the latter(P＜0.05).The results of Pearson test showed that CD4+and CD4+/CD8+were negatively correlated with the prognosis of patients with ankylosing spondylitis(r=-0.568,-0.656,P＜0.001).CD8+was positively correlated with the prognosis of patients with ankylosing spondylitis(r=0.623,P＜0.001).ROC curve analysis showed that the AUC of the combined diagnosis of CD4+,CD8+and CD4+/CD8+for ankylosing spondylitis patients was 0.910,and the diagnostic sensitivity was higher than that of single diagnosis(P＜0.05).Conclusion The abnormal levels of peripheral blood T lymphocyte subsets in patients with ankylosing spondylitis are closely related to the severity and prognosis of the disease,and can be used as a reference indicator for diagnosing the severity and prognosis of ankylosing spondylitis.

【Objective】 To compare the effects of low flux and high flux hemodialysis on insulin resistance (IR), inflammatory factors and coronary artery calcification(CAC) in patients with non-diabetic end-stage renal disease (ESRD), and analyze the related factors affecting the prognosis survival of patients. 【Methods】 A total of 217 patients with non-diabetic ESRD treated in our hospital from February 2015 to April 2017 were selected and randomly divided into control group (n=108) and observation group (n=109) according to the random number table. Low flux and high flux hemodialysis were adopted respectively. Baseline data, renal function, lipid metabolism, inflammatory factors, IR, CAC, complications, outcomes and health and economic benefits of the two groups were compared. The patients were followed up for 3 years and divided into survival group (n=130) and death group (n=75). The clinical data of the two groups were compared and related factors affecting the prognosis and survival were analyzed. 【Results】 Scr, BUN, UAER, TC, TG and LDL-C in the two groups were significantly lower than those before treatment [control group: Scr (μmol/L)349.62±37.16 vs 201.73±24.58, BUN (mmol/L) 28.43±5.39 vs20.81±3.47, UAER(μg /min)60.14±11.52 vs 55.73±9.86, TC (mmol/L)5.46±0.93 vs 4.75±0.69, TG (mmol/L)2.58±0.64 vs 2.13±0.57, LDL-C(mmol/L)3.69±0.73 vs 2.45±0.60; observation group: Scr (μmol/L) 352.14±38.29 vs 136.85±16.47, BUN (mmol/L) 27.96±5.25 vs17.56±3.68, UAER(μg /min) 60.32±12.07 vs 49.85±7.42, TC (mmol/L)5.48±0.97 vs 4.27±0.56, TG (mmol/L) 2.55±0.62 vs 1.49±0.35, LDL-C(mmol/L) 3.72±0.74 vs1.91±0.48), and eGFR and HDL-C were significantly higher than those before treatment [control group: eGFR(mL/min/1.73m)29.32±3.25 vs 72.54±7.86, HDL-C(mmol/L)1.13±0.24 vs1.28±0.31, observation group: eGFR(mL/min/1.73m)30.05±3.29 vs 121.63±13.34, HDL-C(mmol/L)1.09±0.22 vs 1.57±0.46), differences between groups were statistically significant (P<0.05); FBG, FINS, HOMA-IR, IL-6, IL-8, TNF-α and hs-CRP in the two groups were significantly lower than those before treatment [control group: FBG(mmol/L)4.99±0.95 vs 4.52±0.63, FINS(mU/L)12.93±2.54 vs10.15±2.21, HOMA-IR 2.87±0.54 vs 2.04±0.43, IL-6(pg/mL)120.16±13.54 vs 75.94±9.28, IL-8(mg/L)56.83±6.15 vs 41.52±5.38, TNF-α(ng/L)50.03±5.42 vs 45.62±4.81, hs-CRP(mg/L)26.75±2.79 vs 14.37±2.19; observation group: FBG(mmol/L)5.01±0.97 vs 4.11±0.56, FINS(mU/L)13.07±2.62 vs 8.86±1.79, HOMA-IR 2.91±0.55 vs 1.62±0.31, IL-6(pg/mL)119.85±12.91 vs 31.07±4.46, IL-8(mg/L)57.04±6.09 vs 32.65±4.27, TNF-α(ng/L)49.78±5.36 vs 40.15±4.27, hs-CRP(mg/L)23.04±2.82 vs 7.56±1.03], and the CACS score was significantly higher than that before treatment(control group: 26.75±2.79 vs 53.68±26.93, observation group: 27.04±2.82 vs 75.49±7.66), differences between groups are statistically significant (all P<0.05). Compared with the control group, the total incidence of complications during dialysis was significantly lower in the observation group (P<0.05), and has more economic advantages.Venerable age(OR=1.893, P<0.05), low HDL-C level(OR=0.575, P<0.05), high CACS score(OR=2.384, P<0.05), and high hs-CRP level(OR=3.526, P<0.05) were independent risk factors affecting the survival rate of non-diabetic ESRD patients after dialysis treatment (P<0.05). 【Conclusion】 Compared with low-flux hemodialysis, high-flux hemodialysis has significant effects in improving renal function, lipid metabolism, IR, micro-inflammatory state, and reducing CAC progression and complications, with more prominent cost-effectiveness advantages. HDL-C and Hs-CRP levels and CACS scores of patients should be closely monitored during clinical application, and active preventive measures should be taken to improve the survival rate of patients.