Inhibition of valve mesenchymal stromal cell calcium deposition by bFGF through alternative polyadenylation regulation of the CAT gene.

OBJECTIVE: Calcific aortic valve disease (CAVD) is the leading cause of angina, heart failure, and death from aortic stenosis. However, the molecular mechanisms of its progression, especially the complex disease-related transcriptional regulatory mechanisms, remain to be further elucidated. METHODS: This study used porcine valvular interstitial cells (PVIC) as a model. We used osteogenic induced medium (OIM) to induce calcium deposition in PVICs to calcify them, followed by basic fibroblast growth factor (bFGF) treatment to inhibit calcium deposition. Transcriptome sequencing was used to study the mRNA expression profile of PVICs and its related transcriptional regulation. We used DaPars to further examine alternative polyadenylation (APA) between different treatment groups. RESULTS: We successfully induced calcium deposition of PVICs through OIM. Subsequently, mRNA-seq was used to identify differentially expressed mRNAs for three different treatments: control, OIM-induced and OIM-induced bFGF treatment. Global APA events were identified in the OIM and bFGF treatment groups by bioinformatics analysis. Finally, it was discovered and proven that catalase (CAT) is one of the potential targets of bFGF-induced APA regulation. CONCLUSION: We described a global APA change in a calcium deposition model related to CAVD. We revealed that transcriptional regulation of the CAT gene may contribute to bFGF-induced calcium deposition inhibition.

Construction of antibiotic-induced depression mice model and the function of intestinal microbiota.

Many research studies focus on intestinal microbiota-related depression induced by the usage of antibiotics, but the use of antibiotics is fairly different. To construct an effective antibiotic-induced depression mice model and explore the effect of intestinal microbiota in antibiotic-induced depression, we used several kinds of antibiotic mixtures to induce mice depression and used depression-related behavioral tests and neurobiological factors to evaluate the construction of the antibiotic-induced depression mice model. SPSS statistical software was used to analyze the above data, and the optimal model was selected according to the stability of the results and the simplicity of the modeling methods. Metagenomic analysis and fecal microbiota transplantation (FMT) of intestinal microbiota from antibiotic-induced depression mice were performed to analyze the effect of intestinal microbiota. The results showed that antibiotic mixture A (1.25 µg/mL natamycin, 5 mg/mL neomycin sulfate, and 5 mg/mL bacitracin), antibiotic mixture B (24 mg/mL bacitracin, 24 mg/mL neomycin sulfate, 9.6 mg/mL ampicillin, 4.8 mg/mL meropenem, and 1.47 mg/mL vancomycin), and antibiotic solution D (only containing 5 mg/mL neomycin sulfate) could induce depression-like behavior in mice. By using these antibiotics, the concentrations of norepinephrine (NE), 5-hydroxytryptamine (5-HT), and brain-derived neurotrophic factor (BDNF) in mice hippocampus and prefrontal cortex tissues were significantly decreased. All the above results were consistent with those of chronic unpredictable mild stress (CUMS) depression mice. The FMT results showed that fecal microbiota from antibiotic-induced depressed mice transplanted into normal mice (8 weeks-old male C57BL/6J SPF mice) also could induce depression-like behavior and cause similar changes in neurobiological factors. Metagenomic analysis showed that the community structure of microbiota in the intestinal tract of antibiotic-induced depression mice was significantly different from that in control mice, the intestinal microbiota species diversity in antibiotic-induced depression mice was lower, the lipoic acid metabolism pathway was significantly activated, and the abundance of functional gene lipA was explicitly increased. Quantitative real-time PCR (qPCR) further verified the abundance of enriched bacteria in the intestinal microbiota of antibiotic-induced depression mice. In summary, the specific antibiotic mixtures can induce depression by causing changes in intestinal microbiota in mice. Antibiotic-induced depressed mice show differences in intestinal microbiota abundance, high enrichment of the unique metabolic pathway, and the functional gene.

miR-199a-5p inhibits aortic valve calcification by targeting ATF6 and GRP78 in valve interstitial cells.

Calcific aortic valve disease (CAVD) is an important cause of disease burden among aging populations. Excessive active endoplasmic reticulum stress (ERS) was demonstrated to promote CAVD. The expression level of miR-199a-5p in patients with CAVD was reported to be downregulated. In this article, we aimed to investigate the function and mechanism of miR-199a-5p in CAVD. The expression level of miR-199a-5p and ERS markers was identified in calcific aortic valve samples and osteogenic induction by real-time quantitative polymerase chain reaction (RT-qPCR), immunohistochemistry, and western blotting (WB). Alizarin red staining, RT-qPCR, and WB were used for the verification of the function of miR-199a-5p. The dual luciferase reporter assay and rescue experiment were conducted to illuminate the mechanism of miR-199a-5p. In our study, the expression level of miR-199a-5p was significantly decreased in calcified aortic valves and valve interstitial cells' (VICs) osteogenic induction model, accompanying with the upregulation of ERS markers. Overexpression of miR-199a-5p suppressed the osteogenic differentiation of VICs, while downregulation of miR-199a-5p promoted this function. 78 kDa glucose-regulated protein (GRP78) and activating transcription factor 6 (ATF6), both of which were pivotal modulators in ERS, were potential targets of miR-199a-5p. miR-199a-5p directly targeted GRP78 and ATF6 to modulate osteoblastic differentiation of VICs. miR-199a-5p inhibits osteogenic differentiation of VICs by regulating ERS via targeting GRP78 and ATF6.

Effect of preoperative pulmonary artery pressure on the prognosis of end-stage heart failure patients after heart transplantation.

OBJECTIVE: To evaluate the effect of preoperative pulmonary artery pressure on perioperative outcome of end-stage heart failure patients undergoing heart transplantation. METHODS: Retrospective analysis was undertaken on the clinical data of patients receiving heart transplantation in the Department of Cardiovascular Surgery of our hospital from March 2017 to March 2022. A ROC curve analysis was developed between mean pulmonary artery pressure (mPAP) and postoperative mortality using mPAP as diagnostic criteria. Patients were divided into groups based on this threshold to determine the best mPAP threshold value for predicting postoperative nosocomial mortality, and the differences in preoperative and intraoperative data, postoperative complications, and clinical prognosis of patients in the two groups were compared. Patients were followed up to draw the survival curve of patients in the two groups. RESULTS: The study enlisted the participation of 105 patients. ROC curve research revealed that preoperative pulmonary artery pressure was substantially linked with death following heart transplantation, with mPAP = 30.5mmHg being the best threshold. The group with mPAP ≥ 30.5mmHg had a greater incidence of postoperative ECMO support (28.2% vs. 10.6%, P = 0.021) and a higher incidence of in-hospital mortality (15.4% vs. 1.5%, P = 0.019) than the group with mPAP < 30.5mmHg. The postoperative survival rates of 105 patients were 91.3%, 88.7%, 81.6%, and 77.5% at 1, 2, 3, and 4 years, respectively, however, there was no significant difference between the two groups of patients in the postoperative intermediate-far survival rate (P = 0.431). CONCLUSIONS: Preoperative pulmonary artery pressure in patients with end-stage heart failure is intimately correlated with perioperative prognosis of heart transplant recipients. The optimal cut-off mPAP value in predicting perioperative prognosis of heart transplant recipients is 30.5mmHg. In the high mPAP group, perioperative ECMO support rate and perioperative mortality rate are high, which do not affect the medium and long-term prognosis of the recipients undergoing heart transplantation.

Long-term outcomes after aortic valve surgery in patients with aortic regurgitation with preserved ejection fraction and left ventricular dilation.

OBJECTIVES: This study aims to assess the long-term outcomes and prognostic predictors of asymptomatic patients with severe aortic regurgitation (AR) accompanied by left ventricular ejection fraction (LVEF) ≥ 55% and left ventricular end-diastolic diameter (LVEDD) > 65 mm undergoing aortic valve replacement (AVR). METHODS: We retrospectively studied 291 consecutive asymptomatic patients with severe AR accompanied by LVEF ≥ 55% and LVEDD > 65 mm undergoing AVR from January 2000 to December 2013. The long-term outcomes and prognostic predictors were evaluated. RESULTS: There were 2 (0.7%) in-hospital deaths caused by multiple organ failure. The overall survival rate was 95.2% at 5 years, 89.9% at 10 years, 85.9% at 15 years, and 85.9% at 20 years. The left ventricular end-systolic volume index (LVESVi) was an independent predictor of overall mortality, with 59 ml/m2 being the best cut-off value. The left ventricular (LV) dimension decreased within 1 year after surgery and sustained thereafter. There were 15.5% of patients had incomplete LV reverse remodeling. LVESVi was an independent predictor of incomplete LV reverse remodeling, with 56 ml/m2 being the best cut-off value. CONCLUSIONS: AVR can be performed with an acceptable outcome in patients with severe AR accompanied by LVEF ≥ 55% and LVEDD > 65 mm. The LVESVi has the best predictive value for prognosis and the cut-off value is 59 ml/m2, and has the best predictive value for incomplete LV reverse remodeling and the cut-off value is 56 ml/m2.

Mitral valve repair and concomitant maze procedure versus catheter ablation in the treatment of atrial functional mitral regurgitation.

BACKGROUND: To compare mitral valve (MV) repair and concomitant maze procedure with catheter ablation in treating patients with atrial functional mitral regurgitation (AFMR). METHODS: We retrospectively identified 126 patients with AFMR from January 2012 to December 2015. Of these patients, 60 patients underwent MV repair and concomitant maze procedure, and 66 patients received catheter ablation. Patients were followed up for 7.98 ± 2.01 years. The survival, readmission of heart failure (HF), persistent atrial fibrillation (AF), persistent moderate-severe mitral regurgitation (MR) and tricuspid Regurgitation (TR), and echocardiographic data were analyzed in the follow-up. Predictors of readmission of HF were analyzed. RESULTS: There was no significant difference in baseline and echocardiographic characteristics, in-hospital mortality, and other adverse events postoperatively between two groups. The surgical group was associated with lower rates of MR > 2 + grade either at discharge (P = 0.0023) or in the follow-up (P = 0.0001). There was no significant difference in the incidence of overall survival between the two groups. The surgical group was associated with a lower rate of readmission of HF and AF in the follow-up. Univariable and multivariable analysis confirmed AF at discharge, moderate-severe MR at discharge, no MV surgery, moderate-severe TR at discharge, and LA volume as predictors of readmission of HF. Both groups experienced significant reverse cardiac remodeling. CONCLUSIONS: Our results suggest that for the treatment of AFMR with persistent or long-standing persistent AF and moderate-severe MR, MV repair and concomitant maze procedure may achieve a better outcome than catheter ablation procedure.

Glutamic acid assisted hydrolysis strategy for preparing prebiotic xylooligosaccharides.

Since the immune-boosting properties as well as the benefit of promoting the growth of gut bacteria, xylooligosaccharides as prebiotics have attracted considerable interest as functional feed additives around the world. A growing number of studies suggest that acidic hydrolysis is the most cost-effective method for treating xylan materials to prepare xylooligosaccharides, and organic acids were proved to be more preferable. Therefore, in this study, glutamic acid, as an edible and nutritive organic acid, was employed as a catalyst for hydrolyzing xylan materials to prepare xylooligosaccharides. Further, xylooligosaccharide yields were optimized using the response surface methodology with central composite designs. Through the response surface methodology, 28.2 g/L xylooligosaccharides with the desirable degree of polymerization (2-4) at a yield of 40.5 % could be achieved using 4.5% glutamic acid at 163°C for 41 min. Overall, the application of glutamic acid as a catalyst could be a potentially cost-effective method for producing xylooligosaccharides.

Antibiotics-induced depression in mice via the microbiota-gut-brain axis.

BACKGROUND: Intestinal dysbacteriosis is associated with depression. This study aimed to establish an antibiotics-induced depression mouse model and explore the mechanism of antibiotic-induced depression. METHODS: C57BL/6 J mice were treated with antibiotics to prepare the antibiotic-induced depression mouse model. Behavioral tests and depression-related bio-markers were examined. To understand the abundance of different bacteria in intestinal flora and screen out the predominant bacterial species, metagenomic analysis of feces was carried out. Finally, we detected the expression of NF-κB-p65 and p-NF-κB-p65 in PFC and the hippocampus using Western blot. RESULTS: Mixtures A and B caused depression-like behavior in mice. Norepinephrine, 5-hydroxytryptamine, and brain-derived neurotrophic factor in hippocampus and PFC of antibiotic-induced depression mice significantly decreased. The serum adrenocorticotropic hormone and corticosterone concentrations increased. The abundance values of Bacteroides thetaiotaomicron, Klebsiella oxytoca, and Klebsiella aerogenes in antibiotic-induced depression mice significantly increased, and the characteristic KO genes and metabolic pathways in antibiotic-induced depression mice were significantly different with in CUMS depression mice (the positive control) and normal mice. The relative levels of p-NF-κB-p65 in antibiotics-induced depression mice were significantly higher than in normal mice. LIMITATIONS: How dysbacteriosis induces inflammation in the central nervous system is unclear. CONCLUSIONS: Specific antibiotic mixture can cause depression-like behavior and changes of depression-related bio-markers in mice. The antibiotic-induced depression mice display changes in the species and metabolism of intestinal bacterial flora. The activation of NF-κB inflammatory signaling pathway in the central nervous system may act as one of the mechanisms in the development of antibiotic-induced depression.

[The role of intestinal microbiota in tumor occurrence, development and immunotherapy: a review].

The intestinal microbiota is a complex micro-ecological system symbiotic with human body, which has attracted increasing attention in recent years. The intestinal microbiota plays important roles not only in maintaining normal physiological functions of the human body but also in the occurrence, development, diagnosis and treatment of tumors. This review summarized the relationship between the intestinal microbiota and tumor, highlighting the mechanisms by which intestinal microbiota modulates tumor occurrence, development and immunotherapy, particularly the immune checkpoint therapy. This review also summarized the currently available methods for enhancing the efficacy of tumor therapy through regulation of intestinal microbiota. Challenges in the field as well as future perspectives were also discussed.

Production of Prebiotic Xylooligosaccharides via Dilute Maleic Acid-Mediated Xylan Hydrolysis Using an RSM-Model-Based Optimization Strategy.

Xylooligosaccharides (XOS) are functional feed additives that are attracting growing commercial interest owing to their excellent ability to modulate the composition of the gut microbiota. The acid hydrolysis-based processing of xylan-containing materials has been proposed to represent a cost-effective approach to XOS preparation, with organic acids being preferable in this context. As such, in the present study, maleic acid was selected as a mild, edible organic acid for use in the hydrolysis of xylan to produce XOS. A response surface methodology (RSM) approach with a central composite design was employed to optimize maleic acid-mediated XOS production, resulting in a yield of 50.3% following a 15 min treatment with 0.08% maleic acid at 168°C. Under these conditions, the desired XOS degree of polymerization (2-3) was successfully achieved, demonstrating the viability of this using a low acid dose and a high reaction temperature to expedite the production of desired functional products. Moreover, as maleic acid is a relatively stable carboxylic acid, it has the potential to be recycled. These results suggest that dilute maleic acid-based thermal treatment of corncob-derived xylan can achieve satisfactory XOS yields, highlighting a promising and cost-effective approach to XOS production.

Pilot Scale Elimination of Phenolic Cellulase Inhibitors From Alkali Pretreated Wheat Straw for Improved Cellulolytic Digestibility to Fermentable Saccharides.

Depleting supplies of fossil fuel, regular price hikes of gasoline and environmental deterioration have necessitated the search for economic and eco-benign alternatives of gasoline like lignocellulosic biomass. However, pre-treatment of such biomass results in development of some phenolic compounds which later hinder the depolymerisation of biomass by cellulases and seriously affect the cost effectiveness of the process. Dephenolification of biomass hydrolysate is well cited in literature. However, elimination of phenolic compounds from pretreated solid biomass is not well studied. The present study was aimed to optimize dephenoliphication of wheat straw using various alkalis i.e., Ca(OH)2 and NH3; acids i.e., H2O2, H2SO4, and H3PO4; combinations of NH3+ H3PO4 and H3PO4+ H2O2 at pilot scale to increase enzymatic saccharification yield. Among all the pretreatment strategies used, maximum reduction in phenolic content was observed as 66 mg Gallic Acid Equivalent/gram Dry Weight (GAE/g DW), compared to control having 210 mg GAE/g DW using 5% (v/v) combination of NH3+H3PO4. Upon subsequent saccharification of dephenoliphied substrate, the hydrolysis yield was recorded as 46.88%. Optimized conditions such as using 1%+5% concentration of NH3+ H3PO4, for 30 min at 110°C temperature reduced total phenolic content (TPC) to 48 mg GAE/g DW. This reduction in phenolic content helped cellulases to act more proficiently on the substrate and saccharification yield of 55.06% was obtained. The findings will result in less utilization of cellulases to get increased yield of saccharides by hydrolyzing wheat straw, thus, making the process economical. Furthermore, pilot scale investigations of current study will help in upgrading the novel process to industrial scale.

Identification of lncRNA expression profiles and analysis of ceRNA in the hippocampus of perinatal glyphosate-exposed mice.

OBJECTIVE: In order to understand the role of long noncoding RNAs (lncRNAs) played in the mechanisms of glyphosate neurotoxicity in neuronal development. METHODS: Perinatal glyphosate exposure (PGE) mouse model was constructed, and a lncRNA microarray was used to study the lncRNA expression changes in the hippocampus tissue of perinatal glyphosate exposure mice. Then we used GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) databases to analyze the function of the differentially expressed mRNAs and lncRNAs. RESULTS: LncRNA microarray analysis revealed that 1759 lncRNAs and 759 mRNAs were differentially expressed in the perinatal glyphosate exposure (PGE) mice group (G group) compared with the normal control mice group (C group). The functions of the DEmRNAs are involved in the cellular response to hormone stimulus. The ceRNA analysis showed that some interaction networks existed, including (ENSMUST00000137546, ENSMUST00000160950)/(miR-34a-3p, miR-130a-3p)/(Il12b, Irf1). Further analysis of the target mRNAs of miRNAs indicated that the possible functions involved the neuroactive ligand-receptor interaction and calcium signaling pathway, which are involved in perinatal glyphosate exposure-induced neurotoxicity. CONCLUSION: The aberrant expression of lncRNAs is related to the perinatal glyphosate-exposed neurotoxicity. These lncRNAs affect the target gene expression level, might by regulating neuroactive ligand-receptor interactions. The (ENSMUST00000137546, ENSMUST00000160950)/ (miRNA-34a-5p, miR-130a-3p) / mRNAs (e.g., Il12b, Irf1) interaction network may functions in perinatal glyphosate exposure-induced neurotoxicity.

The Evolution of Orbital Implants and Current Breakthroughs in Material Design, Selection, Characterization, and Clinical Use.

It is occasionally essential to surgically remove the damaged eye of the patient in the case of serious oculoorbital injuries, intraocular cancers, and other life-threatening diseases. An orbital implant is placed into the anophthalmic socket after the eye is removed to provide adequate volume reinstatement and revamp the cosmetic look of a normal eye. In the previous few decades, implant design and material selection criteria have progressed from basic nonporous polymeric spheres to devices with more complicated shapes and functions to ensure improved long-term clinical results. Because of their highly interconnected porous design, ceramic and polymeric porous implants have found popularity as a passive framework for fibrovascular ingrowth, with lower obstacle rates and the option of setting to improve prosthetic eye mobility. These materials, however, are not without flaws. The danger of migration and extrusion, infections after surgery, and poor motility transferred to the cosmetic ocular prosthesis are important elements of orbital implants of today. As a result, the development of novel biomaterials with improved functionalities (i.e., antibacterial effect, angiogenesis, and in situ moldability) that allow better eye replacement is more desirable than ever, highlighting one of the most challenging aspects of research topics in the field of ocular implants. This study highlights the history of orbital implants. It gives an outline of current advancements in the area, over and above some essential observations for materials design, selection, characterization, and transformation to clinical applications.

Risk factors of chronic left ventricular dysfunction after cardiac valve surgery.

BACKGROUND: To analyze the risk factors of chronic left ventricular dysfunction (LVD) after cardiac valve surgery. METHODS: A retrospective analysis of 860 patients who underwent heart valve surgery in our center from January 2017 to December 2018, including 650 males and 210 females, aged 58±5.8 years. Inclusion criteria: (I) the patient was clinically diagnosed with heart valve disease and met the surgical indications for mitral valve replacement (MVR), mitral valve repair (MVP), aortic valve replacement (AVR) and double valve replacement (DVR); (II) if atrial fibrillation, coronary artery disease, and tricuspid regurgitation are combined before surgery, radiofrequency ablation, coronary bypass and tricuspid angioplasty were performed contemporarily. Exclusion criteria: (I) preoperative LVEF <50%; (II) aortic dissection underwent Bentall and right heart valve replacement procedures; (III) cardiopulmonary resuscitation and death during perioperative period and 6 months after operation; (IV) postoperative CRRT, IABP, or ECMO assistance; (V) postoperative cardiac dysfunction due to valvular dysfunction, perivalvular leak, or infective endocarditis. Patients were divided into LVD group (LVEF <40%) and control group (LVEF ≥40%) based on cardiac LVEF at 6 months after surgery. Logistic regression was used to analyze the risk factors of postoperative LVD. RESULTS: There were 126 cases in LVD group and 734 cases in control group. There were significant differences in preoperative coronary artery disease, atrial fibrillation, pulmonary hypertension, NYHA classification, left ventricular end diastolic diameter (LVEDD), and left ventricular end systolic diameter (LVESD) between the two groups (P<0.05). The differences in the changes of LVEDD and LVESD before and after operation between the two groups were statistically significant (P<0.05). Logistic regression analysis showed that preoperative LVEDD >55 mm, preoperative LVESD >40 mm, preoperative combined atrial fibrillation, preoperative combined pulmonary hypertension, preoperative NYHA III-IV, and preoperative combined coronary artery disease were the risks of postoperative chronic LVD. CONCLUSIONS: The left ventricular diameter, preoperative coronary artery disease, NYHA III-IV, preoperative atrial fibrillation, and preoperative pulmonary hypertension are risk factors for chronic LVD after heart valve surgery.

Efficacy of cardiovascular surgery for Marfan syndrome patients: a single-center 15-year follow-up study.

BACKGROUND: Most Marfan syndrome (MFS) patients have thoracic aortic diseases which is the major cause of death. The aim of the study is to analyze the impact of different surgical procedures on prognosis of MFS patients. METHODS: We retrospectively analyzed the results of hospitalization and long-term follow-up of MFS patients who underwent surgical intervention in our center. RESULTS: Of the 135 MFS patients, 11 died during hospitalization (8.1%). There were no statistical differences in in-hospital mortality between the proximal surgery group and the distal surgery group (P=0.11). Compared to patients who underwent proximal aortic surgery, patients who underwent arch and distal surgery were more likely to have postoperative respiratory dysfunction (P=0.008). The type of surgical procedure was not associated with the incidence of complications during hospitalization. Pre-surgical New York Heart Association (NYHA) Functional Classification IV (P=0.047), EF <50% (P=0.047), pre-surgical atrial fibrillation (P=0.042), and the injury of dissection propagating onto coronary arteries (P=0.02) were independent risk factors for post-surgical mortality. After 15 years of follow-up, there were no deaths in the David group, while the 15-year survival rate for patients in the Bentall group was 73%±13.5%, and 71%±13.9% for patients in the arch surgery group (P=0.42). The probability of patients in the David group not requiring re-surgery after 15 years was 58.9%±20%, while it was 58.7%±12.1% for patients in the Bentall group, 71.5%±10.5% for patients in the Bentall + Arch group, and 12.5%±11.7% for patients in the Arch + Stent group (P=0.007). CONCLUSIONS: The David procedure was the most beneficial and had the highest long-term patient survival rates.

Epidemiological analysis and rapid detection by one-step multiplex PCR assay of Haemophilus influenzae in children with respiratory tract infections in Zhejiang Province, China.

BACKGROUND: Haemophilus influenzae (H. influenzae) is one of the most important pathogenic bacteria causing respiratory tract infection diseases in children. There are two main types of H. influenzae, encapsulated H. influenzae and nontypeable H. influenzae (NTHi). Serotype b of H. influenzae (Hib) used to be the main epidemic type of H. influenzae, causing the invasive infection. However, the epidemiology of invasive H. influenzae disease has changed substantially, and most invasive diseases are now caused by NTHi and other serotypes of H. influenzae. The aim of this study was to determine the main epidemic strains of H. influenzae in Zhejiang Province in China, and establish a one-step multiplex PCR assay to distinguish H. influenzae from other bacteria associated with respiratory tract infections, and distinguish encapsulated H. influenzae from NTHi. METHOD: In this study, bacterial culture and serum agglutination testing were used to determine the most prevalent serotype of H. influenzae, and the results have served as a gold standard for clinical diagnosis. We also designed a one-step multiplex PCR assay using several kinds of standard strains of respiratory tract infection bacteria, to examine the stability, specificity, and detection limit of the PCR assays. We then used 1514 nasopharyngeal secretion (NPS) samples collected from children with respiratory tract infections to verify the specificity and sensitivity of the PCR assay. RESULTS: The bacterial culture and serum agglutination test results showed that the positive rates of H. influenzae and encapsulated H. influenzae were 18.49 and 1.18%, respectively. The PCR results showed that the detection limit of the multiplex PCR assay was 1.89 × 103 copies /µL, the ompP6 positive rate was 19.35%, and the bexA positive rate was 1.32%. The sensitivity and specificity of the multiplex PCR were 100 and 99.86%, respectively. CONCLUSIONS: According to our study, the most prevalent H. influenzae subtype in Zhejiang Province was NTHi, account for 93.57%; the one-step multiplex PCR assay we established can be used as the differential detection of clinical H. influenzae strains, replacing routine bacterial culture and serum agglutination testing.

Usefulness of interferon-γ release assay for the diagnosis of sputum smear-negative pulmonary and extra-pulmonary TB in Zhejiang Province, China.

BACKGROUND: Quick diagnosis of smear-negative pulmonary tuberculosis (TB) and extra-pulmonary TB are urgently needed in clinical diagnosis. Our research aims to investigate the usefulness of the interferon-γ release assay (IGRA) for the diagnosis of smear-negative pulmonary and extra-pulmonary TB. METHODS: We performed TB antibody and TB-IGRA tests on 389 pulmonary TB patients (including 120 smear-positive pulmonary TB patients and 269 smear-negative pulmonary TB patients), 113 extra-pulmonary TB patients, 81 patients with other pulmonary diseases and 100 healthy controls. Blood samples for the TB-Ab test and the TB-IGRA were collected, processed, and interpreted according to the manufacturer's protocol. RESULTS: The detection ratio of smear-positive pulmonary TB patients and smear-negative pulmonary TB patients were 90.8% (109 of 120) and 89.6% (241 of 269), respectively. There was no statistically significant difference of its performance between these two sample sets (P > 0.05). The detection ratio of positive TB patients and extra-pulmonary TB patients were 90.0% (350 of 389) and 87.6% (99 of 113), respectively, which was not significantly different (P > 0.05). CONCLUSIONS: In this work, the total detection ratio using TB-IGRA was 89.4%, therefore TB-IGRA has diagnostic values in smear-negative pulmonary TB and extra-pulmonary TB diagnosis.

Effect of camptothecin on inducible nitric oxide synthase expression in the colon cancer SW480 cell line.

As a topoisomerase I inhibitor, camptothecin (CPT) is regarded as an effective antitumor agent. In an attempt to search for its novel anticancer mechanism, the present study evaluated the effects of CPT on inducible nitric oxide synthase (iNOS) in the human colon cancer SW480 cell line when stimulated with lipopolysaccharide (LPS) and interleukin (IL)-1ß. The data indicated that CPT significantly decreased NO production. Consistent with these observations, the protein and mRNA expression levels of iNOS were inhibited by CPT in a dose-dependent manner. Thus, the inhibitory effects of CPT on LPS/IL-1ß-stimulated NO production were likely mediated via the inhibition of iNOS gene transcription. From these results, we propose that the inhibition of NO biosynthesis by CPT may partially underlie the efficacy of this antitumor agent.

Human papillomavirus prevalence and type-distribution among women in Zhejiang Province, Southeast China: a cross-sectional study.

BACKGROUND: Human papillomavirus (HPV) infection is the main etiological factor for cervical cancer and premalignant lesions of the cervix. The purposes of the present study were to determine the prevalence of type-specific HPV infections and the association of different HPV types with cervical dysplasia among women in Zhejiang province, Southeast China. METHODS: A total of 15,267 women presenting to a gynaecological outpatient clinic were enrolled in this study. Women were screened for HPV in addition to routine cervical cytology testing. Microarray hybridization and liquid-based cytology tests were used to detect HPV genotypes and cervical cytology, respectively. RESULTS: Based on the population attending a gynaecological outpatient clinic, overall prevalence of any 23 HPV type was 22.8% and multiple HPV infection was found in 4.0% of all the outpatients. HPV prevalence showed bimodal age distribution, with a peak (55.7%) at the ≤20 age group and a second one (35.5%) at >60 age group. In total samples, the five most frequent types were HPV 16 (4.4%), 58 (2.9%), 52 (2.7%), 33 (2.2%) and 11 (1.9%). Overall HPV prevalence increased with the severity of the cytologic result. Analysis through crude odds ratios (ORs) revealed that the cervical lesion risk of HPV-infected women increased to about 26-fold of uninfected women (OR 26.1, 95% CI 22.4 to 30.3). The five most risky HPV types associated with abnormal cytology were HPV 73, 16, 82, 45 and 51. CONCLUSIONS: This study provided baseline data on HPV prevalence in women attending a gynecological outpatient clinic in Zhejiang province. Our data will supply guidance for the primary screening and vaccination program for cervical cancer in this area.

Mechanisms by which the infection of Sclerotinia sclerotiorum (Lib.) de Bary affects the photosynthetic performance in tobacco leaves.

BACKGROUND: Sclerotinia sclerotiorum (Lib.) de Bary is a necrotrophic fungal pathogen which causes disease in a wide range of plants. An observed decrease in photosynthetic performance is the primary reason for the reduction of crop yield induced by S. sclerotiorum. The H2C2O4 is the main pathogenic material secreted by S. sclerotiorum, but the effects of H2C2O4 acidity and the C2O4 2- ion on photosynthetic performance remain unknown. RESULTS: S. sclerotiorum infection significantly decreased photosynthetic O2 evolution and the maximum quantum yield of photosystem II (Fv/Fm) in tobacco leaves under high-light. H2C2O4 (the main pathogenic material secreted by S. sclerotiorum) with pH 4.0 also significantly decreased photosynthetic performance. However, treatment with H3PO4 and HCl at the same pH as H2C2O4 caused much less decrease in photosynthetic performance than H2C2O4 did. These results verify that the acidity of the H2C2O4 secreted by S. sclerotiorum was only partially responsible for the observed decreases in photosynthesis. Treatment with 40 mM K2C2O4 decreased Fv/Fm by about 70% of the levels observed under 40 mM H2C2O4, which further demonstrates that C2O4(2-) was the primary factor that impaired photosynthetic performance during S. sclerotiorum infection. K2C2O4 treatment did not further decrease photosynthetic performance when D1 protein synthesis was fully inhibited, indicating that C2O4(2-) inhibited PSII by repressing D1 protein synthesis. It was observed that K2C2O4 treatment inhibited the rate of RuBP regeneration and carboxylation efficiency. In the presence of a carbon assimilation inhibitor, K2C2O4 2 treatment did not further decrease photosynthetic performance, which infers that C2O4(2-) inhibited PSII activity partly by repressing the carbon assimilation. In addition, it was showed that C2O4(2-) treatment inhibited the PSII activity but not the PSI activity. CONCLUSIONS: This study demonstrated that the damage to the photosynthetic apparatus induced by S. sclerotiorum is not only caused by the acidity of H2C2O4, but also by C2O4(2-) which plays a much more important role in damaging the photosynthetic apparatus. C2O4(2-) inhibits PSII activity, as well as the rate of RuBP regeneration and carboxylation efficiency, leading to the over production of reactive oxygen species (ROS). By inhibiting the synthesis of D1, ROS may further accelerate PSII photoinhibition.