Da-Cheng-Qi decoction improves severe acute pancreatitis capillary leakage syndrome by regulating tight junction-associated proteins.

Background and aims: To investigate mechanisms underlying the effects of Da-Cheng-Qi decoction (DCQD) on severe acute pancreatitis (SAP) capillary leakage syndrome. Methods: In this study, a SAP rat model was established using retrograde perfusion of 5% sodium taurocholate into the biliopancreatic duct. The study included three randomized groups: control, SAP (modeling), and DCQD (via gavage at 2 h pre-modeling and 2 and 4 h post-modeling). HPLC was used to analyzed major components of DCQD. Pathological changes and capillary permeability in the rat pancreatic tissues were examined. mRNA levels of claudin 5, occludin, zonula occludin-1 (ZO-1), and junctional adhesion molecules (JAM-C) were assessed using qRT-PCR. Tight junction-associated protein expression was evaluated using immunofluorescence and Western blot analyses. Human umbilical vein endothelial cells (HUVECs) were used to investigate the mechanism m of DCQD. Results: Serum levels of amylase, TNF-α, IL-1ß, IL-2, and IL-6 were higher in the SAP group compared to the DCQD group (p < 0.05). DCQD treatment significantly attenuated rat pancreas damage (p < 0.05) and reduced tissue capillary permeability compared to the SAP group (p < 0.05). Claudin 5, occludin, and ZO-1 expression in the rat tissues was upregulated, but JAM-C was downregulated by DCQD treatment (p < 0.05). HUVEC permeability was improved by DCQD in a dose-time-dependent manner compared to the SAP group (p < 0.05). DCQD also upregulated claudin 5, occludin, and ZO-1 expression in vitro (p < 0.05). Conclusion: DCQD can improve capillary permeability in both in vivo and in vitro models of SAP by upregulating expression of claudin 5, occludin, and ZO-1, but not JAM-C.

Clinical characterization and proteomic profiling of lean nonalcoholic fatty liver disease.

Introduction: Obesity has been historically associated with nonalcoholic fatty liver disease (NAFLD), but it can also occur in lean individuals. However, limited data is available on this special group. To investigate the clinical and proteomic characteristics of lean subjects with NAFLD, and to identify potential clinical variables and plasma proteins for diagnosing NAFLD in lean individuals, we collected clinical data from a large cohort of 2,236 subjects. Methods: Diagnosis of NAFLD relied on detecting pronounced hepatic steatosis through abdominal ultrasonography. Participants were categorized into four groups based on body mass index: overweight NAFLD, overweight control, lean NAFLD, and lean control. Plasma proteomic profiling was performed on samples from 20 subjects in each group. The lean NAFLD group was compared to both lean healthy and obese NAFLD groups across all data. Results and discussion: The results indicated that the lean NAFLD group exhibited intermediate metabolic profiles, falling between those of the lean healthy and overweight NAFLD groups. Proteomic profiling of plasma in lean subjects with or without NAFLD revealed 45 statistically significant changes in proteins, of which 37 showed high diagnostic value (AUC > 0.7) for lean NAFLD. These potential biomarkers primarily involved lipid metabolism, the immune and complement systems, and platelet degranulation. Furthermore, AFM, GSN, CFH, HGFAC, MMP2, and MMP9 have been previously associated with NAFLD or NAFLD-related factors such as liver damage, insulin resistance, metabolic syndromes, and extracellular homeostasis. Overall, lean individuals with NAFLD exhibit distinct clinical profiles compared to overweight individuals with NAFLD. Despite having worse metabolic profiles than their healthy counterparts, lean NAFLD patients generally experience milder systemic metabolic disturbances compared to obese NAFLD patients. Additionally, the plasma proteomic profile is significantly altered in lean NAFLD, highlighting the potential of differentially expressed proteins as valuable biomarkers or therapeutic targets for diagnosing and treating NAFLD in this population.

Co-infection with human papillomavirus and sexually transmitted infections among Chinese individuals.

OBJECTIVES: This study aimed to investigate the prevalence of Human Papillomavirus (HPV), Sexually Transmitted Infections (STIs) and their co-infections on different genders and ages. METHODS: Different samples of secretions from the reproductive system were collected from 459 males and 494 females for HPV and STI detection. RESULTS: Total HPV infection rate was 49.46 % for males and 48.99 % for females, and the distribution of HPV subtypes varied significantly between different genders. The infection rate of HR-HPV 52 and 31 in females was higher than that in males (p = 0.002 and 0.039, respectively). In contrast, the infection rate of LR-HPV 6 and 11 in males was higher than that in females (p = 0.01 and 0.001, respectively). Females had a significantly higher infection rate of Ureaplasma urealyticum (UU). Besides, these STIs were stratified based on age and the results indicated that the highest incidence of STIs was observed in younger patients (<20 years old). Patients with HPV infections had a higher incidence of Chlamydia trachomatis (CT) in both males and females. CONCLUSIONS: There is a need to perform HPV, CT and UU screening among patients, and more thorough health education for younger patients is of great clinical significance to improve treatment and prognosis.

Virulence is one of the mechanisms of vulvovaginal candidiasis recurrence, rather than drug resistance.

To explore the mechanism of vulvovaginal candidiasis (VVC) recurrence. A total of 127 strains of Candida albicans (C. albicans) were collected, including 58, 40, and 29 strains from the recurrent vulvovaginal candidiasis (RVVC), VVC, and asymptomatic carrier (AC), respectively. The strains' virulence such as in vivo hypha formation rate, germ tube formation rate, biofilm formation ability, and sensitivity to five common antifungals were detected. The in vivo hypha formation rates of C. albicans from the RVVC (55.2%) and VVC (40.0%) were significantly higher than that from the AC (0%) (P < .001). The median germ tube formation rate of the RVVC was 88.2%, which was higher than that of the VVC and AC (59.9% and 65.6%), respectively (P < .001). The median absorbance of the biofilm formation test for strains in the RVVC was 0.380, considerably higher than that in the VVC and AC (0.246 and 0.254) (P < .001). The drug sensitivity rate of the strains to 5-fluorocytosine and itraconazole and the ratio of strains sensitive to all the five antifungals in the VVC group were lower than those in the RVVC and AC groups. In conclusion, the virulence of strains from the RVVC is stronger than that of strains from the VVC and AC, the antifungal resistance rate of strains from the RVVC group is lower than that of strains from the VVC group. So, it is suitable to argue that the strains' virulence is one of the mechanisms for the relapse of RVVC, rather than its antifungal resistance.

The virulence of strains from recurrent vulvovaginal candidiasis (RVVC) is stronger than that from vulvovaginal candidiasis (VVC) and asymptomatic carrier (AC), but the drug resistance rates are opposite. Virulence is one of the mechanisms of the relapse of RVVC, rather than drug resistance.

Genetic diversity and molecular epidemiology of Candida albicans from vulvovaginal candidiasis patients.

Candida albicans (C. albicans) is a common cause of vulvovaginal candidiasis (VVC). In this paper, the genetic diversity and molecular epidemiology of 173C. albicans strains were investigated by multilocus sequence typing (MLST). A total of 52 diploid sequence types (DSTs) were recognized, and 27 (51.9%) of which have not been reported in the MLST database. Genotyping was performed on the multiple isolates collected from patients with recurrent VVC (RVVC, referring to VVC which attacks more than 4 times in one year) in different acute infectious phases. The results showed that 59.1% (26/44) of the patients suffered a relapse, with DST 79 (65.4%) as the dominant genotype. The etiology of the remaining 40.9% (18/44) of patients was reinfection, and the main genotypes included DST 79 (33.3%), DST 124 (8.6%) and DST 1895 (8.6%). DST 79 (45%) and DST 1395 (7.5%) were the main isolates of VVC patients, while DST 79 (24.1%), DST 727 (6.9%), DST 732 (6.9%) and DST 1867 (6.9%) were the main types of healthy volunteers. The results of the genotypes between RVVC patients and other groups were statistically different. Furthermore, cluster analysis was carried out on 1468 isolates, among which 1337 were downloaded from the MLST database, 130 were divided into 8 Clades in the present study and the remaining one was taken as a singleton. 92.3% isolates from relapse patients, 58.3% isolates from re-infected patients, 77.5% isolates from VVC patients and 51.7% isolates from volunteers were distributed in Clade 1. The analysis of the genotypes of multiple isolates from RVVC patients further demonstrated that point mutation and loss of heterozygosity contributed to the microevolution of C. albicans.

Recurrent tuberculosis in an urban area in China: Relapse or exogenous reinfection?

Recurrent tuberculosis is an important indicator of the effectiveness of tuberculosis control and can occur by relapse or exogenous reinfection. We conducted a retrospective cohort study on all bacteriologically confirmed tuberculosis cases that were successfully treated between 2000 and 2012 in Shanghai, an urban area with a high number but a low prevalence rate of tuberculosis cases and a low prevalence of HIV infection. Genotyping the Mycobacterium tuberculosis from clinical isolates was used to distinguish between relapse and reinfection. In total, 5.3% (710/13,417) of successfully treated cases had a recurrence, a rate of 7.55 (95% CI 7.01-8.13) episodes per 1000 person-years, more than 18 times the rate of tuberculosis in the general population. Patients who were male, age 30-59, retreatment cases, had cavitation, diabetes, drug-resistant or multidrug-resistant tuberculosis in their initial episode of tuberculosis, were at high risk for a recurrence. Among 141 recurrent cases that had paired isolates, 59 (41.8%) had different genotypes, indicating reinfection with a different strain. Patients who completed treatment were still at high risk of another episode of tuberculosis and exogenous reinfection contributed a significant proportion of the recurrent tuberculosis cases. Targeted control strategies are needed to prevent new tuberculosis infections in this setting.

Transmission of multidrug-resistant Mycobacterium tuberculosis in Shanghai, China: a retrospective observational study using whole-genome sequencing and epidemiological investigation.

BACKGROUND: Multidrug-resistance is a substantial threat to global elimination of tuberculosis. Understanding transmission patterns is crucial for control of the disease. We used a genomic and epidemiological approach to assess recent transmission of multidrug-resistant (MDR) tuberculosis and identify potential risk factors for transmission. METHODS: We did a population-based, retrospective study of patients who tested positive for tuberculosis between Jan 1, 2009, and Dec 31, 2012, in Shanghai, China. We did variable-number-of-tandem-repeat genotyping and whole-genome sequencing of isolates. We measured strain diversity within and between genomically clustered isolates. Genomic and epidemiological data were combined to construct transmission networks. FINDINGS: 367 (5%) of 7982 patients with tuberculosis had MDR tuberculosis and 324 (88%) of these had isolates available for genomic analysis. 103 (32%) of the 324 MDR strains were in 38 genomic clusters that differed by 12 or fewer single nucleotide polymorphisms (SNPs), indicating recent transmission of MDR strains. Patients who had delayed diagnosis or were older than 45 years had high risk of recent transmission. 235 (73%) patients with MDR tuberculosis probably had transmission of MDR strains. Transmission network analysis showed that 33 (87%) of the 38 clusters accumulated additional drug-resistance mutations through emergence or fixation of mutations during transmission. 68 (66%) of 103 clustered MDR strains had compensatory mutations of rifampicin resistance. INTERPRETATION: Recent transmission of MDR tuberculosis strains, with increasing drug-resistance, drives the MDR tuberculosis epidemic in Shanghai, China. Whole-genome sequencing can measure of the heterogeneity of drug-resistant mutations within and between hosts and help to determine the transmission patterns of MDR tuberculosis. FUNDING: National Science and Technology Major Project, National Natural Science Foundation of China, and US National Insitutes of Health.

Cartilage Oligomeric Matrix Protein-Angiopoietin-1 Has a Protective Effect of Vascular Endothelial Barrier in Rat With Acute Necrotizing Pancreatitis.

OBJECTIVE: To investigate the protective effect of angiopoietin-1 (Ang-1) from capillary endothelial damage in rats with acute necrotizing pancreatitis (ANP). METHODS: 96 male Sprague-Dawley rats were randomly averaged and divided into control group, ANP group, Si-Ang-1 group, and COMP (cartilage oligomeric matrix protein)-Ang-1 group. Animals were killed at 6, 12, and 24 hours after molding. Levels of serum amylase, porcine endothelin 1, C-reactive protein, and Ang-1 were detected; histopathological changes in the pancreas were observed; capillary permeability and Ang-1 expression of the pancreatic tissue were detected by Evans Blue extravasation assay, immunohistochemistry, Western blot, and quantitative polymerase chain reaction. RESULTS: (1) Levels of serum amylase, C-reactive protein, and porcine endothelin-1 increased and level of Ang-1 decrease in the ANP group and Si-Ang-1 group compared with the control group, whereas COMP-Ang-1 group could improve the changes. (2) The order of pancreas pathological changes (mild to severe) is: control group, COMP-Ang-1 group, ANP group, and Si-Ang-1 group. (3) Capillary permeability of the pancreatic tissue in the COMP-Ang-1 group was lower than that in the ANP group. (4) Ang-1 mRNA and protein expression in the COMP-Ang-1 group was significantly higher than in the ANP group. CONCLUSIONS: COMP-Ang-1 can upregulate the expression of Ang-1 protein to promote angiogenesis and improve early inflammatory and pathological damage in ANP group.

[Application of melting curve assay in the quality assessment of drug susceptibility test on Mycobacterium tuberculosis].

OBJECTIVE: To evaluate the feasibility of melting curve assay technologies in quality assessment of drug susceptibility test (DST) on Mycobacterium tuberculosis. METHODS: During May 2009 to September 2012, a total of 2 204 MTB isolates were collected in Shanghai Songjiang District and 5 other county level TB institutes. DST results of these isolates were verified by the melting curve assay. Isolates with inconsistent phenotype and genotype results were retested and sequenced for any drug resistance mutations. RESULTS: Evaluation at Songjiang District showed high consistency of the melting curve assay with rifampicin and isoniazid DST results (kappa = 0.97 and 0.99, respectively). Consistency of the two methods were 97.6% (41/42), 100% (757/757) and 99.9% (788/799) in rifampicin resistant, susceptible, and total cases, respectively. As for isoniazid resistance detection, the consistency were 100% (72/72), 99.9% (726/727), and 99.9% (798/799) in resistant, susceptible, and total cases, respectively. Blinded re-examination of isolates with inconsistent genotype and phenotype resistance confirmed the good reproducibility of melting curve assay, which yielded the same results as the first test. Quality assessment at institutes A,B, C,D and E, however, showed consistency of only 87.5% (21/24), 80.8% (21/26), 87.5% (14/16), 82.9% (29/35), 60.9% (14/23) in rifampicinresistant cases, and 88.2% (30/34), 65.6% (21/32), 85.0% (17/20), 68.3% (41/60), 62.5% (15/24) in isoniazid resistant cases. As the reliability of DST results significantly varied among different areas, our finding indicated that there were misdiagnosed drug resistant cases in some field sites. CONCLUSION: The melting curve assay for MTB DST is fast, easy to use, reproducible, and able to identify mistakes in daily work. Therefore, it is a suitable tool for the quality assessment of conventional DST results.

Down-regulation of aquaporin 1 in rats with experimental acute necrotizing pancreatitis.

OBJECTIVES: The objectives of this study were to investigate the expression of aquaporin 1 in capillary endothelial cells of rats with experimental acute necrotizing pancreatitis (ANP) and to explore its pathogenic role in capillary leak. METHODS: Sixty-four male Sprague-Dawley rats were randomly divided into control (n = 32) and ANP groups (n = 32). Eight rats in each group were killed at 3, 6, 12, and 18 hours after induction of experimental models. Quantity of ascites and levels of serum amylases were measured. Capillary permeability in pancreas, lung, and intestinal tissue was detected by Evans blue extravasation method. Aquaporin 1 expression in pancreas, lung, and intestinal tissue was detected by real-time polymerase chain reaction, immunohistochemical staining, and Western blot. RESULTS: Serum amylase level was significantly higher in ANP group than in controls (P < 0.05). Evans blue concentration in tissues in the ANP group was significantly higher than that in controls (P < 0.05). Aquaporin 1 mRNA and protein expressions in tissues were significantly less in the ANP group than in the controls (P < 0.05). CONCLUSIONS: The expression of aquaporin 1 was down-regulated in the pancreas, lung, and intestinal tissue of ANP rats, which could play an important role in the pathogenesis of capillary leak syndrome.