Mutations in troABCD against Copper Overload in a copA Mutant of Streptococcus suis.

Streptococcus suis is a major swine pathogen that is increasingly recognized as a porcine zoonotic pathogen that threatens the health of both pigs and humans. Metal homeostasis plays a critical role during the process of bacterial infection. In this study, RNA sequencing was used to identify potential candidate genes involved in the maintenance of intracellular copper homeostasis. CopA was identified as the primary copper exporter in S. suis. The copA deletion mutant strain was found to be more sensitive to copper and accumulated more intracellular copper than the wild-type (WT) parent strain. In addition, adding manganese increased the ability of S. suis to resist copper, and the manganese transporter, TroABCD, was involved in tolerance to copper. The copA deletion mutant strain accumulated less copper when supplemented with manganese. Furthermore, when cultured with copper, the double deletion mutant (ΔcopAΔtroA) exhibited improved growth compared to the copA deletion mutant strain. In addition, the double deletion mutant (ΔcopAΔtroA) accumulated less copper than the copA deletion mutant strain. These data were consistent with a model wherein defective TroABCD resulted in decreased cellular copper accumulation and protected the strain against copper poisoning. IMPORTANCE Metal homeostasis plays a critical role during the process of bacterial infection. We identified three important potential candidate genes involved in maintenance of intracellular copper homeostasis. CopA was demonstrated to be the main copper exporter in Streptococcus suis, and manganese increased the tolerance of S. suis to copper. The double deletion mutant (ΔcopAΔtroA) improved growth ability over the copA deletion mutant strain in the presence of high concentrations of copper and accumulated less copper. These findings are consistent with a model wherein defective TroABCD resulted in decreased cellular accumulation of copper and protected the strain against copper poisoning.

The novel effectiveness of Tai Chi on cardiopulmonary fitness among stroke patients in the recovery phase: a study protocol for a randomized controlled trial.

BACKGROUND: Stroke is the leading cause of death worldwide. China faces a similar risk of stroke as developed countries because of considerable changes in lifestyle, such as overeating and smoking. Tai Chi is a traditional form of mind-body exercise that has been widely practiced in China for thousands of years. However, there are few studies on the effect of Tai Chi on the cardiopulmonary function of stroke patients in the recovery phase. Therefore, it is necessary to observe the effect of Tai Chi on the cardiorespiratory fitness of patients after stroke. METHODS: This is a parallel-design, two-arm, analyst assessor-blinded, randomized controlled trial. A total of 226 stroke patients in the recovery phase will be recruited and assigned randomly to a control group or Tai Chi group at a 1:1 ratio. The patients in the Tai Chi group will perform the Tai Chi exercise. The patients in the control group will perform walking exercises. Patients in both groups will receive conventional treatments and healthy education. The primary outcomes will be VO2peak and scores on the MOS item short form health survey (SF-36) scale. Secondary outcomes will include vital capacity (VC), ejection fractions (EF), and cardiac output (CO). The assessments of the tests will be performed at three time points (before exercise, at the end of exercise, and 6 weeks after exercise). Adverse events will be recorded faithfully during the study. DISCUSSION: If the results are positive, this study will contribute to the establishment of further guided Tai Chi rehabilitation programs. TRIAL REGISTRATION: Chinese Clinical Trial Registry ChiCTR2000034719 . Registered on 16 July 2020.

CopA Protects Actinobacillus pleuropneumoniae against Copper Toxicity.

Actinobacillus pleuropneumoniae is a Gram-negative bacterium causing porcine pleuropneumonia and severe economic losses in the global swine industry. The toxic trace element copper is required for many physiological and pathological processes in organisms. However, CopA, one of the most well-characterized P-type ATPases contributing to copper resistance, has not been characterized in A. pleuropneumoniae. We used quantitative PCR analysis to examine expression of the copA gene in A. pleuropneumoniae and investigated sequence conservation among serotypes and other Gram-negative bacteria. Growth characteristics were determined using growth curve analyses and spot dilution assays of the wild-type strain and a △copA mutant. We also used flame atomic absorption spectrophotometry to determine intracellular copper content and examined the virulence of the △copA mutant in a mouse model. The copA expression was induced by copper, and its nucleotide sequence was highly conserved among different serotypes of A. pleuropneumoniae. The amino acid sequence of CopA shared high identity with CopA sequences reported from several Gram-negative bacteria. Furthermore, the △copA mutant exhibited impaired growth and had higher intracellular copper content compared with the wild-type strain when supplemented with copper. The mouse model revealed that CopA had no influence on the virulence of A. pleuropneumoniae. In conclusion, these results demonstrated that CopA is required for resistance of A. pleuropneumoniae to copper and protects A. pleuropneumoniae against copper toxicity via copper efflux.

The mechanism of Suzi-Jiangqi Decoction in treatment of COPD based on network pharmacology / 国际中医中药杂志

Objective:By using the network pharmacology method to predict the active constituents and action targets of Suzi-Jiangqi Decoction in the treatment of COPD, and to explore its potential molecular mechanism with multi-component, multi-target and multi-pathway characteristics. Methods:The active constituents and targets of Suzi-Jiangqi Decoction were collected, screened and predicted according to the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and UniProt software. Search for the gene targets related to COPD in GeneCards, online human Mendelian genetic database (OMIM) and TTD database. The intersection targets of component targets and disease targets were obtained by Veen map online software. The network model with the sequence of active constituents-target-disease was constructed and analyzed by Cytoscape software, and the protein-protein interaction network (PPI) was constructed by STRING database. The gene ontology function annotation (GO) and Tokyo genome encyclopedia (KEGG) pathway enrichment analysis of common targets with metascape online tool. Results:A total of 163 active constituents of Suzi-Jiangqi Decoction were screened, 283 targets were predicted, and 159 targets involved in the treatment of COPD. Quercetin, kaempferol, naringin and luteolin were the key active ingredients. IL6, TNF, MAPK3, JUN, CASP3, CXCL8, CXCL10, MMP9 and MAPK1 were important gene targets. GO analysis showed that the biological processes involved in the enrichment of key gene targets included the response to bacteria, the cytokine mediated signaling pathway, the cell's response to inorganic substances, the response to oxidative stress, the response to LPS, and so on. The enrichment analysis of KEGG pathway showed that the signaling pathway of Suzi-Jiangqi Decoction in the treatment of COPD included TNF signaling pathway, IL-17 signaling pathway, cell cycle, Influenza A, HTLV-I infection, AGE-RAGE signaling pathway, Tuberculosis, Epstein-Barr virus infection and so on. Conclusion:Suzi-Jiangqi Decoction can treat COPD through multi-target and multi-pathway mechanisms of anti-inflammatory, anti infection and immune regulation, which lays a foundation for further study of its molecular mechanism.

A comparison of graph- and kernel-based -omics data integration algorithms for classifying complex traits.

BACKGROUND: High-throughput sequencing data are widely collected and analyzed in the study of complex diseases in quest of improving human health. Well-studied algorithms mostly deal with single data source, and cannot fully utilize the potential of these multi-omics data sources. In order to provide a holistic understanding of human health and diseases, it is necessary to integrate multiple data sources. Several algorithms have been proposed so far, however, a comprehensive comparison of data integration algorithms for classification of binary traits is currently lacking. RESULTS: In this paper, we focus on two common classes of integration algorithms, graph-based that depict relationships with subjects denoted by nodes and relationships denoted by edges, and kernel-based that can generate a classifier in feature space. Our paper provides a comprehensive comparison of their performance in terms of various measurements of classification accuracy and computation time. Seven different integration algorithms, including graph-based semi-supervised learning, graph sharpening integration, composite association network, Bayesian network, semi-definite programming-support vector machine (SDP-SVM), relevance vector machine (RVM) and Ada-boost relevance vector machine are compared and evaluated with hypertension and two cancer data sets in our study. In general, kernel-based algorithms create more complex models and require longer computation time, but they tend to perform better than graph-based algorithms. The performance of graph-based algorithms has the advantage of being faster computationally. CONCLUSIONS: The empirical results demonstrate that composite association network, relevance vector machine, and Ada-boost RVM are the better performers. We provide recommendations on how to choose an appropriate algorithm for integrating data from multiple sources.

Characterization of Anthocyanins in Perilla frutescens var. acuta Extract by Advanced UPLC-ESI-IT-TOF-MSn Method and Their Anticancer Bioactivity.

The anthocyanin extract from a domestic Perilla cultivar (Perilla frutescens var. acuta) were isolated and characterized with high mass accuracy and multi-dimensional fragmentation by means of ultra-performance liquid chromatography (UPLC) and electrospray ionization-ion trap-time of flight mass spectrometry analysis (ESI-IT-TOF-MSn). The new developed and applied LC-MS method focused on in-depth screening of anthocyanin compounds with similar structures which also provided a new approach of anthocyanin characterization without the use of external standards. Selective detection of interested anthocyanins was achieved utilizing extracted ion chromatogram (EIC) analysis, while MSn spectra were recorded to allow identification of the anthocyanin based on characteristic fragmentation patterns. Seven anthocyanins including one feruloyl (Cyanidin 3-O-feruloylglucoside-5-O-glucoside), two caffeoyl (Cyanidin 3-O-caffeoylglucoside-5-O-glucoside, Cyanidin 3-O-caffeoylglucoside-5-O-malonylglucoside) and four coumaroyl substituted anthocyanins (Cis-shisonin, Malonyl-cis-shisonin, Shisonin, and Malonyl-shisonin) were identified. Annexin-V FITC/PI flow cytometric assay was performed to analyze the influence of anthocyanin extract of P. frutescens var. acuta on cell apoptosis. The results suggested that Perilla anthocyanins can induce Hela cell apoptosis by a dose dependent manner.

Clinical observations of sequential therapy with Chinese medicine and hysteroscopic mechanical stimulation of the endometrium in infertile patients with repeated implantation failure undergoing frozen-thawed embryo transfer / 中国结合医学杂志

<p><b>OBJECTIVES</b>To investigate the clinical effect of sequential therapeutic intervention Yupei Qisun [compensating for weakness by invigorating Kidney (Shen) and Spleen (Pi) in advance] in Chinese medicine (CM) and hysteroscopic endometrial mechanical stimulation on the treatment of infertile patients with repeated implantation failure (RIF); and to study the differences in patients' endometrial thickness and type on the day of embryo transfer, serum hormone levels on embryo transfer day and clinical pregnancy outcomes.</p><p><b>METHODS</b>In the clinical study, 168 frozen-thawed embryo transfer (FET) cycles for couples with RIF conforming to the research protocol were randomly divided into three groups: a CM group with 56 cycles (CM combined with FET), a hysteroscopy group with 55 cycles (hysteroscopic endometrial mechanical stimulation), and a control group with 57 cycles (conventional FET). Differences in endometrial thickness on the embryo transfer day, levels of serum estradiol (E2) and progesterone (P) on the embryo transfer day, the E2/P ratio on the embryo transfer day, biochemical and clinical pregnancy rates, implantation rate, abnormal pregnancy rate and other indices were compared among the three groups.</p><p><b>RESULTS</b>Endometrial thickness, E2 and P levels, and the E2/P ratio on embryo transfer day and other factors had no significant differences among groups. The biochemical pregnancy, clinical pregnancy, and implantation rates of the CM and hysteroscopy groups were significantly higher than the control group (P<0.05), and there were no significant differences between these two groups. The abnormal pregnancy rate had no significant difference among the three groups.</p><p><b>CONCLUSIONS</b>Sequential therapy of Yupei Qisun could significantly improve the clinical outcomes of rif-fet cycles, being equivalent to hysteroscopic endometrial mechanical stimulation, and provided a reliable method to treat such infertile couples.</p>

Analysis of selenium content in hair samples of children in Kaschin-Beck Disease areas in Gansu Province / 中国地方病学杂志

Objective To compare selenium content in hair samples of children in Kashin-Beck Disease (KBD) areas and control areas,and to provide academic data for KBD control and prevention.Methods In 2011,Longnan,Qingyang,Dingxi City of KDB areas in Gansu Provines were selected as survey point,meanwhile,colleagues of Tianshui City and Lanzhou City were selected as control.Hair samples of children aged between 6 and 12 years were taken from each survey point to determine the hair selenium.After using the nitricperchloric mixed acid to digest the sample,the concentrations of selenium were determined by hydride generator atomic fluorescence spectrometry.Results Two hundred and fifty-four samples from KBD areas and 102 samples from control areas were collected.The hair selenium of KBD areas[(0.22 ± 0.07),(0.22 ± 0.00),(0.20 ± 0.07)mg/kg],were higher than the control areas[(0.32 ± 0.08),(0.42 ±0.11)mg/kg].After comparation of selenium content in every sampling point,we found the range of children's selenium content was relatively narrow in KBD areas (0.08-0.46 mg/kg).The minimum value and the maximum value in KBD areas were lower than that of control areas (0.15-0.51 mg/kg).A totle of 51.1％ (182/356) of children was seriously lack of selenium,the hair selenium content was ＜ 0.25 mg/kg.It was also found that Se content was not significantly different between boys and girls.Conclusions Children's selenium is deficient in KBD areas in Gansu.It is important to ensure salt supplement with selenium for a long period of time and through other ways to increase Selenium intake to control KBD.

Application of Early Enteral Nutrition on Treatment of Severe Acute Pancreatitis / 中国普外基础与临床杂志

Objective To discuss the feasibility and safety of early enteral nutrition (EN) on treatment of severe acute pancreatitis (SAP) and its influence. Methods The advancement about application of early EN on treatment of SAP in recent years were reviewed. Results In patients with SAP, early EN via catheter placed in the jejunum could protect the integrity of intestinal barrier and reduce infectious complications. But no consensus had been reached about the starting time, ingredient and infusion mode yet. Conclusion Early EN may have positive effects on treatment of SAP, but further researches are still needed.