Objective:To construct the rpoN gene knockout strain (Δ rpoN) and the complemented strain (CΔ rpoN) of Vibrio vulnificus ( V. vulnificus), and investigate the role of the rpoN gene in regulating bacterial motility and biofilm formation. Methods:The Δ rpoN strain of V. vulnificus was constructed using homologous recombination. Bacterial motility was assessed via swimming assays, and flagellar morphology was observed by transmission electron microscopy. Biofilm formation capacity was evaluated using crystal violet and Congo red staining assays, as well as colony morphology analysis. Real-time quantitative RT-PCR (qRT-PCR) was used to detect mRNA levels of target genes associated with flagellar synthesis and biofilm formation in Δ rpoN and the wild-type strains. Results:The V. vulnificus genome harbored a single rpoN gene, encoding a protein with high amino acid sequence similarity to RpoN homologs in other bacterial species. The Δ rpoN strain was successfully constructed. Compared with the wild-type strain, the Δ rpoN strain exhibited complete loss of motility on soft agar plates, absence of flagellar, and downregulated mRNA levels of flagellar synthesis-related genes. Conclusions:In V. vulnificus, RpoN regulates flagellar assembly by modulating the expression of flagellar synthesis genes, thereby controlling bacterial motility and biofilm formation.

Background:Liver cirrhosis is characterized by chronic inflammation,progressive fibrosis,and eventual liver dysfunction,and poses a major global health challenge.Aims:To assess the global burden of liver cirrhosis and its risk factors from 1990 to 2021.Methods:Using data extracted from the Global Burden of Diseases,Injuries,and Risk Factors Study(GBD)2021,the incidence,mortality,disability-adjusted life years(DALYs),and their age-standardized rates of liver cirrhosis were analyzed.Furthermore,a stratified analysis was conducted by sex,age,region,and etiology,with projections of the trends in the next 15 years.Results:Compared to 1990,the global incidence number of liver cirrhosis in 2021 was increased by 58.2%,the death number and DALYs rose by 39.5%and 27.9%,respectively.While the global age-standardized incidence rate(ASIR)showed a slight increase,the age-standardized death rate(ASDR)and DALY rate continued to decline.Both ASIR and ASDR exhibited negative correlations with the sociodemographic index(SDI).All age-standardized rates were higher in males than in females.Since 1990,the incidence rate increased in younger populations,while the mortality and DALY rates declined in most age groups.Non-alcoholic fatty liver disease(NAFLD)emerged as the leading cause of incidence,whereas chronic hepatitis B and C remained the primary contributors to deaths and DALYs.The incidence of NAFLD was prominent in high and high-middle SDI regions,while chronic hepatitis B was concentrated in low SDI regions.Projections to 2036 indicated a continuing rise in ASIR,and declines in ASDR and DALY rate.The incidence of chronic hepatitis B was projected to decrease markedly,whereas that of NAFLD was expected to continue increasing.Conclusions:Between 1990 and 2021,the global incidence of liver cirrhosis showed a modest increase;in contrast,both mortality and DALY rates demonstrated a steady decline.Burden of liver cirrhosis poses notable regional disparities.NAFLD dominates incidence in high-income regions,while viral hepatitis remains predominant in low-income areas,highlighting the need for region-specific prevention strategies.

Objective:To construct the rpoN gene knockout strain (Δ rpoN) and the complemented strain (CΔ rpoN) of Vibrio vulnificus ( V. vulnificus), and investigate the role of the rpoN gene in regulating bacterial motility and biofilm formation. Methods:The Δ rpoN strain of V. vulnificus was constructed using homologous recombination. Bacterial motility was assessed via swimming assays, and flagellar morphology was observed by transmission electron microscopy. Biofilm formation capacity was evaluated using crystal violet and Congo red staining assays, as well as colony morphology analysis. Real-time quantitative RT-PCR (qRT-PCR) was used to detect mRNA levels of target genes associated with flagellar synthesis and biofilm formation in Δ rpoN and the wild-type strains. Results:The V. vulnificus genome harbored a single rpoN gene, encoding a protein with high amino acid sequence similarity to RpoN homologs in other bacterial species. The Δ rpoN strain was successfully constructed. Compared with the wild-type strain, the Δ rpoN strain exhibited complete loss of motility on soft agar plates, absence of flagellar, and downregulated mRNA levels of flagellar synthesis-related genes. Conclusions:In V. vulnificus, RpoN regulates flagellar assembly by modulating the expression of flagellar synthesis genes, thereby controlling bacterial motility and biofilm formation.

Background:Liver cirrhosis is characterized by chronic inflammation,progressive fibrosis,and eventual liver dysfunction,and poses a major global health challenge.Aims:To assess the global burden of liver cirrhosis and its risk factors from 1990 to 2021.Methods:Using data extracted from the Global Burden of Diseases,Injuries,and Risk Factors Study(GBD)2021,the incidence,mortality,disability-adjusted life years(DALYs),and their age-standardized rates of liver cirrhosis were analyzed.Furthermore,a stratified analysis was conducted by sex,age,region,and etiology,with projections of the trends in the next 15 years.Results:Compared to 1990,the global incidence number of liver cirrhosis in 2021 was increased by 58.2%,the death number and DALYs rose by 39.5%and 27.9%,respectively.While the global age-standardized incidence rate(ASIR)showed a slight increase,the age-standardized death rate(ASDR)and DALY rate continued to decline.Both ASIR and ASDR exhibited negative correlations with the sociodemographic index(SDI).All age-standardized rates were higher in males than in females.Since 1990,the incidence rate increased in younger populations,while the mortality and DALY rates declined in most age groups.Non-alcoholic fatty liver disease(NAFLD)emerged as the leading cause of incidence,whereas chronic hepatitis B and C remained the primary contributors to deaths and DALYs.The incidence of NAFLD was prominent in high and high-middle SDI regions,while chronic hepatitis B was concentrated in low SDI regions.Projections to 2036 indicated a continuing rise in ASIR,and declines in ASDR and DALY rate.The incidence of chronic hepatitis B was projected to decrease markedly,whereas that of NAFLD was expected to continue increasing.Conclusions:Between 1990 and 2021,the global incidence of liver cirrhosis showed a modest increase;in contrast,both mortality and DALY rates demonstrated a steady decline.Burden of liver cirrhosis poses notable regional disparities.NAFLD dominates incidence in high-income regions,while viral hepatitis remains predominant in low-income areas,highlighting the need for region-specific prevention strategies.

The traditional Chinese medicine （TCM） in pediatric care has a long history，proven efficacy，and distinctive characteristics.The China Association of Chinese Medicine has organized a series of youth salons to discuss the clinical advantages of treating diseases.Experts at this seminar proposed that the superior disease categories in pediatric TCM are significant for showcasing the unique strengths and advantages of TCM in the treatment of pediatric diseases，enhancing diagnostic and treatment levels，inheriting TCM knowledge，promoting the integration of TCM and Western medicine，and meeting patients' needs.The strengths of pediatric TCM are summed up as "having what others do not，excelling at what others have，being special in what others excel at，and ensuring safety in what others specialize in." The scope of superiority in pediatric TCM covers multiple systems，including respiratory，digestive，endocrine，psychological，and dermatological systems.This article summarized the advantages of TCM in treating 13 diseases discussed in the salon，such as upper respiratory tract infections，coughs，pneumonia，allergic rhinitis，bronchial asthma，atopic dermatitis，functional dyspepsia，functional constipation，enuresis，marginal short stature，simple obesity，attention deficit hyperactivity disorder，and tic disorders.The overall advantages were reflected in three aspects：First，the holistic concept and treatment based on syndrome differentiation in TCM highlight the advantage of treating the root causes of diseases，making the treatment methodical and precise.Second，most traditional Chinese herbs are natural and have strong safety profiles.Third，TCM treatment methods are abundant，especially the external treatment methods，which are widely used in pediatrics，highly accepted by parents，and have definite efficacy，as well as good safety and compliance.

Objective To assess the efficacy of pirfenidone combined with PD-L1 inhibitor for treatment of bladder cancer in a mouse model and its effect on tumor immune microenvironment modulation.Methods Forty C57BL/6 mouse models bearing ectopic human bladder cancer xenografts were randomized into control group,PD-L1 inhibitor group,pirfenidone group and combined treatment group(n=10).After successful modeling,PD-L1 inhibitor treatment was administered via intraperitoneal injection at 12.5 mg/kg every 3 days,and oral pirfenidone(500 mg/kg)was given on a daily basis.The survival rate of the mice and tumor growth rate were compared among the 4 groups.The expressions of CD3,CD8,CD45,E-cadherin and N-cadherin in the tumor tissues were detected with immunohistochemistry after the 21-day treatment,and bone marrow-derived suppressor cells(MDSCs)were observed with immunofluorescence staining;serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),urea nitrogen(BUN),creatinine(CRE)and lactate dehydrogenase(LDH-L)were analyzed using an automated biochemical analyzer.Results Treatment with PD-L1 inhibitor and pirfenidone alone both significantly decreased tumor growth rate and tumor volume at 21 days(P<0.05),but the combined treatment produced an obviously stronger inhibitory effect(P<0.05).PD-L1 inhibitor and pirfenidone alone significantly increased E-cadherin expression and decreased N-cadherin expression in the tumor tissue(P<0.05).The two treatments both significantly increased the percentage of CD3+,CD8 and CD45+ T cells and decreased the percentage of Ly-6G+CD11b+MDSCs in the tumor tissue,and these changes were more obvious in the combined treatment group(P<0.05).No significant differences were found in serum ALT,AST,BUN,CRE or LDH-L levels among the 4 groups(P>0.05).Conclusion Combined treatment with pirfenidone and PD-L1 inhibitor significantly inhibits the progression of bladder cancer in mice possibly by regulating tumor immune microenvironment and inhibiting epithelial-mesenchymal transition of the tumor cells.

Objective To assess the efficacy of pirfenidone combined with PD-L1 inhibitor for treatment of bladder cancer in a mouse model and its effect on tumor immune microenvironment modulation.Methods Forty C57BL/6 mouse models bearing ectopic human bladder cancer xenografts were randomized into control group,PD-L1 inhibitor group,pirfenidone group and combined treatment group(n=10).After successful modeling,PD-L1 inhibitor treatment was administered via intraperitoneal injection at 12.5 mg/kg every 3 days,and oral pirfenidone(500 mg/kg)was given on a daily basis.The survival rate of the mice and tumor growth rate were compared among the 4 groups.The expressions of CD3,CD8,CD45,E-cadherin and N-cadherin in the tumor tissues were detected with immunohistochemistry after the 21-day treatment,and bone marrow-derived suppressor cells(MDSCs)were observed with immunofluorescence staining;serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),urea nitrogen(BUN),creatinine(CRE)and lactate dehydrogenase(LDH-L)were analyzed using an automated biochemical analyzer.Results Treatment with PD-L1 inhibitor and pirfenidone alone both significantly decreased tumor growth rate and tumor volume at 21 days(P<0.05),but the combined treatment produced an obviously stronger inhibitory effect(P<0.05).PD-L1 inhibitor and pirfenidone alone significantly increased E-cadherin expression and decreased N-cadherin expression in the tumor tissue(P<0.05).The two treatments both significantly increased the percentage of CD3+,CD8 and CD45+ T cells and decreased the percentage of Ly-6G+CD11b+MDSCs in the tumor tissue,and these changes were more obvious in the combined treatment group(P<0.05).No significant differences were found in serum ALT,AST,BUN,CRE or LDH-L levels among the 4 groups(P>0.05).Conclusion Combined treatment with pirfenidone and PD-L1 inhibitor significantly inhibits the progression of bladder cancer in mice possibly by regulating tumor immune microenvironment and inhibiting epithelial-mesenchymal transition of the tumor cells.

Objective To study and compare the oral microbiota and metabolites of children with Henoch Schonlein purpura(HSP)to identify specific microbiota and metabolites related to this disease and elucidate the pathogenesis of HSP.Methods Three groups of qualified subjects were included,including 20 in the HSP group,20 in the HSP nephritis(HSPN)group,and 20 in the control group.Perform high-throughput 16S rRNA sequencing and metabolic profiling of saliva from each group to analyze the correlation between differential microbiota and differ-ential metabolites.Results(1)Compared with the control group,there was a significant difference in richness and diversity in the HSPN group(P＜0.05).At the same time,there was no significant difference in richness and diver-sity in the HSP group(P＞0.05).Compared with the HSP group,the abundance,and diversity of the HSPN group were significantly increased(P＜0.05).At the genus level,the proportion of Streptococcus in each group is the high-est.Compared with the control group,there was no significant correlation between the HSP group and the genus of bacteria.In contrast,the HSPN group showed a significant increase in the genera of Pseudomonas and Parabacteroi-des(P＜0.05).Compared with the HSP group,the abundance of Pseudomonas and Parabacteroides in the HSPN group was significantly increased(P＜0.05).(2)Compared with the control group,the HSPN group had 12 differen-tial metabolites involving nine metabolic pathways,such as phenylalanine metabolism;There was no significant dif-ference in metabolites and no metabolic pathway in the HSP group.Compared with the HSP group,the HSPN group has 15 differential metabolites involving nine metabolic pathways,such as phenylalanine metabolism.(3)In the HSPN and control groups,Pseudomonas and Parabacteroides negatively correlated with Phenylalanine metabolic pathway products.In the HSPN and HSP groups,Pseudomonas,Parabacteroides,and Phenylalanine metabolic path-way products were negatively correlated.The metabolites involved in phenylalanine metabolism in the oral cavity are 2-hydroxycinnamic acid,Phenylpyruvic acid,and N-acetyl-L-phenylalanine.Conclusion There is a significant dif-ference between HSPN and HSP children and healthy children.Streptococcus,Pseudomonas,and Parabacteroides may be one of the trigger factors of HSPN,and Phenylalanine metabolism may be one of the pathways in the patho-genesis of HSPN.Children with HSPN have a more pronounced imbalance in oral microbiota and greater differences in metabolic products than children with HSP.

Objective:To investigate the safety and effectiveness of tension adjustment technique using anatomical landmarks during retropubic midurethral synthetic sling.Methods:The data of 36 consecutive female patients with urinary incontinence, who had underwent retropubic midurethral synthetic sling procedure from January to August 2019 were analyzed retrospectively. The mean age was (60.83±7.93) years old and the body mass index was (24.43±2.44) kg/m 2. Among the recruited subjects, 36 had positive stress test and Marshall-Marchetti test. 20 (55.6%) were pure stress urinary incontinence, and 16 (44.4%) were mixed urinary incontinence. The severity of incontinence was classified into mild (5 cases, 13.9%), moderate (14 cases, 38.9%), severe (13 cases, 36.1%) and very severe (4 cases, 11.1%) using one-hour pad tests. Urodynamics were performed in 17 cases, with 5 (29.4%) presented detrusor overactivity, 3 (17.7%) possessed intrinsic sphincter deficiency. For each case, the tension of the sling was adjusted based on the anatomical landmarks, i. e. using an angled clamp attached closely to the pubic symphysis ventrally and the tip parallel to the edge of hymen dorsally. All patients were catheter-free right after the procedure. The subjective and objective effectiveness, and safety (the rate of urinary retension after surgery and postvoid residual volume 3 months later) were evaluated.The subjective cure rate was was defined as complete leakage free or very mild leakage during excessive bladder filling and fierce cough. The subjective effectiveness was defined as over 50% improvement of the leakage symptom. The objective cure rate was defined as a negative stress test. Results:For all 36 patients, the median hospital stays was 8 (5-95)h. No bladder perforation or transfusion cases. All patients were catheter-free right after the procedure, with no incidence of urinary retention. 27 patients completed a 3-month follow-up, with 22 had post-void residual data, 23 had subjective effectiveness data and 23 had objective effectiveness data. The median post-void residual was 7.5 (5-64) ml, subjective cure rate was 91.3% (21/23), and objective cure rate was 95.7% (22/23). 8.7% (2/23) reported difficult urination alleviated without the necessity of clinical interference. No urethra erosion or vagina extrusion was found. At 2-year follow-up, 34 patients completed assessment by phone. The subjective cure rate was 91.2% (31/34), with only 2.9% (1/34) reported difficult urination. Besides, at 3-month follow-up, there was no difference regarding the subjective cure rate [100.0%(12/12) vs. 81.8%(9/11)]or objective cure rate [91.7%(11/12) vs. 100.0%(11/11)] between patients with stress and mixed incontinence. No difference was noted among patients with mild, moderate, severe and very severe leakage[75.0% (3/4) vs. 100.0%(6/6) vs. 90.0%(9/10) vs. 100.0%(3/3)]. Of the 12 cases with urodynamic records, the presence of detrusor overactivity [66.7%(2/3) vs. 88.9%(8/9)] or intrinsic sphincter deficiency [0(0/1) vs. 90.9%(10/11)] did not significantly affected the cure rate of the procedure. At 2-year follow-up, there was no difference regarding the subjective cure rate between patients with stress and mixed incontinence [94.7%(18/19) vs. 86.7%(13/15)]. No difference was also noted among patients with mild, moderate, severe and very severe leakage[80.0%(4/5) vs. 100.0%(13/13) vs. 83.3%(10/12) vs. 100.0%(4/4)]. Of the 16 cases with urodynamic records, the presence of detrusor overactivity [60.0%(3/5) vs. 90.9%(10/11)]or intrinsic sphincter deficiency [66.7%(2/3) vs. 84.6%(11/13)]did not significantly affected the cure rate of the procedure.Conclusions:Tension adjustment using anatomic landmarks during sling procedure is safe and feasible for urinary incontinence, with minimum complications and residual volume, and high subjective/objective cure rate.

Objective:To study the effect of duodenal papillary morphology on selective biliary cannulation (SBC).Methods:Data of 912 patients with biliopancreatic diseases treated with endoscopic retrograde cholangiopancreatography (ERCP) at the Department of Gastroenterology, Affiliated Hospital of Northwest Minzu University from January 2018 to January 2020 were retrospectively analyzed. Duodenal papillary morphology of patients was classified into 4 types by using Haraldsson's endoscopic classification. The success rate of SBC, the pre-incision rate, the difficult intubation rate and the incidence of post-ERCP pancreatitis (PEP) of different papillary types were analyzed.Results:A total of 912 patients were enrolled in this study, and 86.95% (793/912) duodenal papilla conformed to one type of the classification, of which 77.18% (612/793) were regular type (type Ⅰ), 10.21% (81/793) small nipple type (type Ⅱ), 8.58% (68/793) protruding or drooping type (type Ⅲ), and 4.04% (32/793) wrinkled or ridged type (type Ⅳ). The success rates of SBC in four types of duodenal papilla were 98.86% (605/612), 90.12% (73/81), 88.24% (60/68) and 96.60% (28/32) respectively. The success rate of SBC of type Ⅰ was higher than those of type Ⅱ-Ⅳ (all P<0.008) and there was no significant difference between those of type Ⅱ and type Ⅲ ( P> 0.008). The pre-incision rate in the four types of duodenal papilla were 7.84% (48/612), 32.10% (26/81), 50.00% (34/68) and 25.00% (8/32) respectively. The pre-incision rate of type Ⅰ was lower than those of type Ⅱ-Ⅳ (all P<0.008), and there was no significant difference between those of type Ⅱ and type Ⅲ ( P>0.008). The difficult intubation rate in the four types of duodenal papilla were 12.42% (76/612), 39.51% (32/81), 58.82% (40/68) and 28.12% (9/32) respectively .The difficult intubation rate of type Ⅰ was lower than those of type Ⅱ and type Ⅲ (both P<0.001), and that of type Ⅲ was higher than that of type Ⅳ ( P=0.004) . There was no significant difference between those of type Ⅰ and type Ⅳ or type Ⅱ and type Ⅲ (both P>0.008). The incidences of PEP of the four types were 2.61% (16/612), 12.35% (10/81), 5.88% (4/68) and 6.25% (2/32) respectively. The incidences of PEP of type Ⅱ was higher than that of type Ⅰ ( P<0.001) , and there was no significant difference between those of type Ⅰ and type Ⅳ or type Ⅱ and type Ⅲ (both P>0.008). Conclusion:SBC is affected by duodenal papilla morphology, easiest for type Ⅰ and hard for type Ⅱ and Ⅲ. Attention should be paid to risk of PEP in SBC of type Ⅱ.