In order to develop a transgenic zebrafish line with green fluorescent protein (enhanced green fluorescent protein, EGFP) expressed specifically in muscle and heart, the recombinant expression vector constructed using the zebrafish ttn.2 gene promoter fragment and EGFP gene coding sequence and the capped mRNA of Tol2 transposase were co-injected into the zebrafish 1-cell stage embryos. The stable genetic Tg (ttn.2: EGFP) transgenic zebrafish line was successfully developed by fluorescence detection, followed by genetic hybridization screening and molecular identification. Fluorescence signals and whole-mount in situ hybridization showed that EGFP expression was located in muscle and heart, the specificity of which was consistent with the expression of ttn.2 mRNA. Inverse PCR showed that EGFP was integrated into chromosomes 4 and 11 of zebrafish in No. 33 transgenic line, while integrated into chromosome 1 in No. 34 transgenic line. The successful construction of this fluorescent transgenic zebrafish line, Tg (ttn.2: EGFP), laid a foundation for the research of muscle and heart development and related diseases. In addition, the transgenic zebrafish lines with strong green fluorescence can also be used as a new ornamental fish.
Animals ; Zebrafish/genetics* ; Animals, Genetically Modified/genetics* ; Green Fluorescent Proteins/metabolism* ; Zebrafish Proteins/genetics* ; Promoter Regions, Genetic

Objective To reveal the genetic diversity of cultivating resources of Trichosanthes sp. from different areas. Methods The genetic background of 11 populations (including 30 samples) of Trichosanthis Fructus were researched by both ITS and SRAP molecular markers. The software mega 5.0 and NTSY-pc 2.1 were taken to analysis the ITS sequences and SRAP polymorphic bands data respectively. Results There are obvious mutation in ITS sequences among T. kirilowii and T. rosthornii, ITS sequences could authenticate the different resources in some extent. 165 SRAP bands were amplified by using the optimized 15 primer pairs, and 136 were polymorphic bands, the polymorphic rate was up to 82.4%. The samples were separated into two clusters at the genetic similarity coefficient of 0.18, one cluster gathered the samples of Yuelou Num.8 (T. kirilowii) and the other cluster further separated into three sub-clusters at the coefficient of 0.90, which were mainly T. rosthornii. Conclusion There are several species of Trichosanthes have been used to harvest the seeds in China, and the major one is T. rosthornii. The cultivating resources of T. rosthornii have little difference in genetic backgrounds.

Antimicrobial resistance is on the rise while the number of antibiotics being brought to market continues to drop. Drug-resistant genes and drug-resistant bacteria infection have seriously threatened human health. Therefore, antimicrobial resistance presents an ongoing challenge that requires multifaceted approaches including: biomedical innovation; improved surveillance of antibiotic consumption and antimicrobial resistance generated rates; prevention of health-care-associated infections and transmission of multidrug-resistant bacteria and environmental dissemination; rapid microbiological diagnosis; and curtailed clinical and veterinary misuse. Fortunately, combating antimicrobial resistance has been highly valued and supported by the government, scientists and entrepreneurs of various countries. With the continuous introduction of new technologies, new products, and new management measures, the problem of antimicrobial resistance must be controlled and alleviated.

Rotator cuff tear is a common disease that causes shoulder pain and limitation of activity. Rotator cuff repair with arthroscope has become the mainstream method with advance in surgical techniques. In spite of this, the highly rate of retear after surgery mainly because the tendon to bone interface form scar tissue rather than native tissue. Biomechanical of the interface is so poor to retear because of the change of Histological. In recent years, more and more researchers are devoted to the study of biology and biomechanical for improving the process of tendon to bone healing and restoring the original structure at the interface, This article reviews the research progress with four factors(inflammation, slow or limited bone ingrowth into the tendon graft, Mechanical stimulation, scant stem cell) and physiotherapy that affect tendon to bone healing.
Arthroplasty ; Humans ; Rotator Cuff ; Rotator Cuff Injuries ; Tendon Injuries ; Tendons ; Wound Healing

OBJECTIVE：To stu dy the improvement effect s of Dantaojin （Salvia miltiorrhiza ，Persicae Semen ，Curcumae Radix）extract on oxidative stress of liver and kidney in chronic lead poisoning model mice. METHODS ：Totally 72 mice were randomly divided into normal control group ，model group ，positive control group （dimercaptosuccinate，70 mg/kg），Dantaojin extract low-dose ，medium-dose and high-dose groups （20，40，60 g/kg），with 12 mice in each group. Except for normal control group，other groups were given intraperitoneal injection of lead acetate solution 20 mg/kg（every other day ，consecutive 20 days） to establish chronic lead poisoning model. After modeling ，administration groups were given relevant medicine intragastrically ， normal control group and model group were given constant volume of water intragastrically ，once a day ，for consecutive 20 days. After last medication ，body weight of mice was weighed ，and organ coefficients （kidney and liver ）were calculated. Serum levels of ALT ，AST，BUN and Scr were determined by automatic biochemical detector. HE staining was used to observe histopathological features of liver and kidney. ELISA method was used to determine the levels of GSH-Px ，SDO and MDA in liver and renal tissue. RESULTS：Compared with normal control group ，body weight （except for high-dose group ），the levels of SOD （except for high-dose group ）and GSH-Px were all decreased significantly in model group and Dantaojin extract groups ，while the renal coefficients（except for high-dose group ），liver coefficients （except for low-dose ，medium-dose and high-dose group ），the levels of BUN （except for high-dose group ），Scr，AST（except for high-dose group ），ALT and MDA were increased significantly （P＜ 0.05 or P＜0.01）. The epithelial cells of glomerulus and renal tubules were atrophied ，the arrang ement of hepatocytes was loose and some cells were necrotic. Compared with model group ，body weight ，the levels of SOD and GSH-Px were increased significantly in positive control group and Dantaojin extract groups，while the renal and liver coefficients ，the levels of BUN，Scr，AST，ALT and M DA were decreased significantly（P＜0.05 or P＜0.01）. Histopathological fea tures of liver and renal tissue were improved significantly in Dantaojin extract medium-dose and high-dose groups. CONCLUSIONS ：Dantaojin extract could improve oxidant stress injury in liver and renal tissue，the mechanism of which may be associated with eliminating reactive oxygen radicals ，inhibiting lipid peroxidation and enhancing antioxidant defense ability.

OBJECTIVE: To analyze the predictors of successful weaning off extracorporeal membrane oxygenation (ECMO) after extracorporeal cardiopulmonary resuscitation (ECPR). METHODS: The clinical data of 56 patients with cardiac arrest who underwent ECPR in Hunan Provincial People's Hospital (the First Affiliated Hospital of Hunan Normal University) from July 2018 to September 2022 were retrospectively analyzed. According to whether ECMO was successfully weaning off, patients were divided into the successful weaning off group and the failed weaning off group. The basic data, duration of conventional cardiopulmonary resuscitation (CCPR, the time from cardiopulmonary resuscitation to ECMO), duration of ECMO, pulse pressure loss, complications, and the use of distal perfusion tube and intra-aortic balloon pump (IABP) were compared between the two groups. Univariate and multivariate Logistic regression analyses were performed to identify the risk factors for weaning failure of ECMO. RESULTS: Twenty-three patients (41.07%) were successfully weaned from ECMO. Compared with the successful weaning off group, patients in the failed weaning off group were older (years old: 46.7±15.6 vs. 37.8±16.8, P < 0.05), higher incidence of pulse pressure loss and ECMO complications [81.8% (27/33) vs. 21.7% (5/23), 84.8% (28/33) vs. 39.1% (9/23), both P < 0.01], and longer CCPR time (minutes: 72.3±19.5 vs. 54.4±24.6, P < 0.01), shorter duration of ECMO support (hours: 87.3±81.1 vs. 147.7±50.8, P < 0.01), and worse improvement in arterial blood pH and lactic acid (Lac) levels after ECPR support [pH: 7.1±0.1 vs. 7.3±0.1, Lac (mmol/L): 12.6±2.4 vs. 8.9±2.1, both P < 0.01]. There were no significant differences in the utilization rate of distal perfusion tube and IABP between the two groups. Univariate Logistic regression analysis showed that the factors affecting the weaning off ECMO of ECPR patients were pulse pressure loss, ECMO complications, arterial blood pH and Lac after installation [pulse pressure loss: odds ratio (OR) = 3.37, 95% confidence interval (95%CI) was 1.39-8.17, P = 0.007; ECMO complications: OR = 2.88, 95%CI was 1.11-7.45, P = 0.030; pH after installation: OR = 0.01, 95%CI was 0.00-0.16, P = 0.002; Lac after installation: OR = 1.21, 95%CI was 1.06-1.37, P = 0.003]. After adjusting for the effects of age, gender, ECMO complications, arterial blood pH and Lac after installation, and CCPR time, showed that pulse pressure loss was an independent predictor of weaning failure in ECPR patients (OR = 1.27, 95%CI was 1.01-1.61, P = 0.049). CONCLUSIONS Early loss of pulse pressure after ECPR is an independent predictor of failed weaning off ECMO in ECPR patients. Strengthening hemodynamic monitoring and management after ECPR is very important for the successful weaning off ECMO in ECPR.
Humans ; Extracorporeal Membrane Oxygenation ; Blood Pressure ; Retrospective Studies ; Perfusion ; Cardiopulmonary Resuscitation

Genome sequences of 4 644 representative strains from human gut microbiota were analyzed to mine gene clusters for biosynthesis of novel secondary metabolites, as well as genes encoding antibiotic resistance and virulence factors. AntiSMASH analysis showed that more than 60% of the representative strains encoded at least one secondary metabolite gene cluster, and 8 potential novel secondary metabolite gene clusters were identified from 8 unculturable bacteria. The secondary metabolite gene clusters in human intestine are mainly composed of nonribosomal peptide synthetase (NRPS), bacteriocin, arylpolyene, terpene, betalactone and NRPS like gene clusters distributed in Clostridia, Bacilli, Gammaproteobacteria, Bacteroidia, Actinobacteria and Negativicutes. PathoFact analysis showed that genes encoding antibiotic resistance and virulence factors are widely distributed in representative strains, but the frequency encoded by potential pathogens is significantly higher than that of non-potential pathogens. The frequency of genes encoding secretory toxins such as outer membrane protein, PapC N-terminal domain, PapC C-terminal domain, peptidase M16 inactive domain, and non-secretory toxins such as nitroreductase family, AcrB/AcrD/AcrF family, PLD-like domain, Cupin domain, putative hemolysin, S24-like peptidase, phosphotransferase enzyme family, endonuclease/ exonuclease/ phosphatase family, glyoxalase/ bleomycin resistance was high in potential pathogens. This study may facilitate mining new microbial natural products from the intestinal microbiome, understanding the colonization and infection mechanism of intestinal microorganisms, and providing targeted prevention and treatment of intestinal microbial related diseases.
Humans ; Virulence ; Multigene Family ; Bacteria ; Drug Resistance, Microbial ; Virulence Factors ; Peptide Hydrolases

Objective To investigate and predict the molecular targets and mechanism of Huanglian Jiedu Decoction (黄连解毒汤, HLJDD) in the treatment of Corona Virus Disease 2019 (COVID-19) through network pharmacology and molecular docking analysis. Methods The chemical constituents and action targets of HLJDD were retrieved on Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), SymMap v2, Encyclopedia of Traditional Chinese Medicine (ETCM), a High-throughput Experiment- and Reference-guided Database of Traditional Chinese Medicine (HERB), and Traditional Chinese Medicine Integrated Database (TCMID). UniProt and GeneCards were used to query the target genes that corresponding to the active compounds, and then a compound-target network was constructed using Cytoscape 3.7.2. Gene Ontology (GO) database was used to annotate GO functions. Kyoto Encyclopedia of Genes and Genomes (KEGG) was used to predict the possible mechanisms of active compounds. The Database for Annotation, Visualization and Integrated Discovery (DAVID) was used to analysis the tissue enrichment. The main active compounds in HLJDD are molecularly docked with their corresponding related targets. Results Seventy-six compounds were screened and 458 corresponding targets in the network were obtained. Gene annotation showed that the targets were involved mainly in 1 953 biological processes. 884 signaling pathways was enriched, involving signaling by interleukins, cytokine signaling in immune system, generic transcription pathway, and RNA polymerase II transcription. The targets mainly distributed in the lung, liver, and placenta, involving a variety of immune cells, such as T cells and B cells. The molecular docking results showed that core compounds such as wogonin, berberine, and baicalein had high affinity with tumor necrosis factor (TNF), insulin (INS), and tumor protein 53 (TP53). Conclusion The active compounds in HLJDD may have a therapeutic effect on COVID-19 through regulating multiple signal pathways by targeting genes such as vascular endothelial growth factor A (VEGFA), INS, interleukin-6 (IL-6), TNF, caspase-3 , TP53, and mitogen-activated protein kinase 3 (MAPK3).

OBJECTIVETo study on the effect of acupuncture at early stage on prognosis of the infantile of anoxic and ischemic cerebral injury.

METHODSOne hundred and forty-eight newborns of moderate and severe anoxic and ischemic encephalopathy after acute stage were randomly divided into an acupuncture group and a control group. The acupuncture group were treated with comprehensive interference with acupuncture as main after the acute stage, and the control group with a routine infanticulture measure after acute stage and were discharged from hospital. Intelligence test was conducted in age of 3, 6, 12 and 18 months. Results The average development quotient (DQ) in the: acupuncture group in age of 6, 12, 18 months increased by 14.54, 21.46, and 24.06 points as compared with the control group, respectively. After age of 18 months, the maim rate was 7.2% in the treatment group and 33.8% in the control group with a very significant difference between the two groups (P < 0.01).

CONCLUSIONThe comprehensive interference with acupuncture as main at early stage can effectively promote mental and motion development, and it is effective way for improving prognosis, reducing incidence rate of sequelae and increasing life quality in the infantile of anoxic and ischemic encephalopathy.

Objective: To examine whether and how Viticis Fructus total flavonoids (VFTF) inhibit the capacity of self-renewal in lung cancer stem cells (LCSCs) derived from human small cell lung cancer NCI-H446 cell line. Methods: NCI-H446 cell line was cultured in vitro. LCSCs were obtained and amplified by Magnetically activated cell sorting system (MACS) and suspended culture with serum-free conditioned medium. The capacity of self-renewal was detected by tumor sphere-forming assay. The protein expression levels of the self-renewal associated transcription factors, Bmi1 and p-Akt, were analyzed using Western blotting. Results: VFTF significantly suppressed the capacity of self-renewal in LCSCs, in a concentration-dependent manner (P < 0.05). The expression levels of Bmi1 and p-Akt were elevated in LCSCs, compared with parental cells. VFTF effectively down-regulated the expression levels of Bmi1 and p-Akt in LCSCs. In addition, LY294002, a PI3K specific inhibitor, enhanced the inhibition of VFTF on the capacity of self-renewal in LCSCs. Conclusion: VFTF could inhibit the self-renewal capacity of LCSCs derived from NCI-H446 cell line, which is associated with down-regulation of the expression of p-Akt and self-renewal associated transcription factors Bmi1.