Traditional Chinese medicine for gastric cancer: An evidence mapping.

As a complementary and alternative therapy, traditional Chinese medicine (TCM) has been playing a significant role in gastric cancer treatment. Data from individual systematic reviews have not been comprehensively summarized, and the relationship between certain interventions and outcomes are ill-defined. This study aimed to analyze the advantages of TCM interventions for gastric cancer by the method of evidence mapping. We searched PubMed, Embase, Web of Science, China National Knowledge Infrastructure, Chinese Scientific Journals Database, and Wanfang Database for systematic reviews of TCM treating gastric cancer up to December 31, 2023. We used Excel, Endnote 20, and Python software for the analysis of incorporated studies. We assessed the quality of included SRs by AMSTAR-2 and performed evidence mapping including 89 SRs, 1648 RCTs and 122,902 patients, identifying 47 types of interventions and 39 types of outcomes. From a visual overview, we displayed that most SRs reported beneficial effects in improving short- and long-term survival, myelosuppression, and immune function, even though the quality of evidence was generally low. The benefits of Brucea javanica Oil Emulsion Injection, ShenQiFuZheng Injection, XiaoAiPing, Astragalus-Containing TCM and Guben Xiaoji Therapy were found the most solid in corresponding aspects. Our findings suggest that although more rigorous clinical trials and SRs are needed to identify the precise effectiveness, integrating such evidence into clinical care of gastric cancer is expected to be beneficial.

Thiazolopyrimidinone Derivative H5-23 Enhances Daptomycin Activity against Linezolid-Resistant Enterococcus faecalis by Disrupting the Cell Membrane.

The increasing emergence and dissemination of multidrug-resistant (MDR) Gram-positive pathogens pose a serious threat to global public health. Previous reports have demonstrated that the compound H5-23, which has a thiazolopyrimidinone core structure, exhibited antibacterial activity against Staphylococcus epidermidis in vitro. However, the antibacterial activity in vivo and mechanism of action of H5-23 against MDR bacteria have not been fully studied. In this study, we report that H5-23 has wide-spectrum antibacterial activity against Gram-positive bacteria. When combined with daptomycin (DAP), H5-23 demonstrates enhanced antimicrobial activity, effectively killing both planktonic and persister cells, as well as eradicating biofilm formation by linezolid-resistant Enterococcus faecalis. The development of resistance shows that H5-23 has a low propensity to induce antibiotic resistance compared to that of linezolid in vitro. Mechanistic studies reveal that H5-23 increases membrane permeability and disrupts membrane integrity, resulting in increased production of reactive oxygen species (ROS), metabolic perturbations, and ultimately cell death. Additionally, we demonstrate the synergistic antibacterial effect of H5-23 combined with DAP in a murine model. These findings suggest that H5-23 is a promising antimicrobial agent and provides a potential strategy for enhancing the efficacy of DAP in combating multidrug-resistant E. faecalis.

Surgical site infection following open lobectomy in patients with lung cancer: A prospective study.

OBJECTIVE: To assess the incidence and risk factors of surgical site infection after open pulmonary lobectomy and to quantify their clinical and economic burden. METHODS: A prospective nested case-control study was performed on patients with lung cancer who underwent open lobectomy in the lung cancer center of West China Hospital from January 2017 to December 2019. Demographic, clinical data and medical costs were recorded. Logistic regression was used to evaluate risk factors associated with surgical site infection. A Mann-Whitney U test was carried out to evaluate the differences in medical costs. RESULTS: A total of 1395 patients were eligible, and the surgical site infection incidence was 13.47% (188/1395). Of the 188 instances of surgical site infection, 171 (90.96%) were classified as organ/space infection, 8 (4.25%) as superficial incisional infection and 9 (4.79%) as deep incisional infection. The patients with surgical site infection had significantly higher mortality (3.19% vs. 0.41%, p < 0.001), higher median medical cost (90774.95 yuan vs. 63079.38 yuan, p < 0.001), and longer postoperative length of stay (15 days vs. 9 days, p < 0.001). Multivariate logistic regression analysis indicated that age (odds ratio (OR) = 1.560, p = 0.007), respiratory failure (OR = 5.984, p = 0.0012), American Society of Anesthesiologists score (OR = 1.584, p = 0.005), operating time (OR = 1.950, p < 0.001), and operation team (OR = 1.864, p < 0.001) were independent risk factors for surgical site infection. CONCLUSIONS: The high incidence of surgical site infection indicates that postoperative infections remain a significant clinical burden in patients who underwent open lobectomy. Identifying risk factors timely through prospective surveillance may assist clinical decisions against surgical site infection.

The potential target of bithionol against Staphylococcus aureus: design, synthesis and application of biotinylated probes Bio-A2.

This study aims to explore the potential targets of bithionol in Staphylococcus aureus.The four bithionol biotinylated probes Bio-A2-1, Bio-A2-2, Bio-A2-3, and Bio-A2-4 were synthesized, the minimal inhibitory concentrations (MICs) of these probes against S. aureus were determined. The bithionol binding proteins in S. aureus were identified through immunoprecipitation and LC-MS/MS with bithionol biotinylated probe. The biotinylated bithionol probes Bio-A2-1 and Bio-A2-3 displayed antibacterial activities against S. aureus. The Bio-A2-1 showed lower MICs than Bio-A2-3, and both with the MIC50/MIC90 at 12.5/12.5 µM against S. aureus clinical isolates. The inhibition rates of bithionol biotinylated probes Bio-A2-1 and Bio-A2-3 on the biofilm formation of S. aureus were comparable to that of bithionol, and were stronger than that of Bio-A2-2 and Bio-A2-4. The biofilm formation of 10 out of 12S. aureus clinical isolates could be inhibited by Bio-A2-1 (at 1/4×, or 1/2× MICs). There are three proteins identified in S. aureus through immunoprecipitation and LC-MS/MS with bithionol biotinylated probe Bio-A2-1: Protein translocase subunit SecA 1 (secA1), Alanine--tRNA ligase (alaS) and DNA gyrase subunit A (gyrA), and in which the SecA1 protein the highest coverage and the most unique peptides. The LYS112, GLN143, ASP213, GLY496 and ASP498 of SecA1 protein act as hydrogen acceptors to form 6 hydrogen bonds with bithionol biotinylated probe Bio-A2-1 by molecular docking analysis. In conclusion, the bithionol biotinylated probe Bio-A2-1 has antibacterial and anti-biofilm activities against S. aureus, and SecA1 was probably one of the potential targets of bithionol in S. aureus.

Total Synthesis of Immunosuppressive Mycestericin E and G Enabled by a Highly Stereoselective Nitroso-Ene Cyclization.

This report describes a streamlined synthesis of immunosuppressive mycestericin E and G through a highly stereoselective nitroso-ene cyclization in 11-12 steps using readily available materials. The stereochemical outcome in the formation of a Nα-quaternary stereogenic center is rationalized by a trajectory based on the polar diradical intermediate and subsequent hydrogen transfer. Julia olefination offers a facile chain elongation method that presents a viable strategy for structural derivatization in future medicinal applications.

Early extracorporeal membrane oxygenation as bridge for central airway obstruction patients caused by neck and chest tumors to emergency surgery.

Central airway obstruction caused by neck and chest tumors is a very dangerous oncological emergency with high mortality. Unfortunately, there is few literature to discuss an effective way for this life-threating condition. Providing effective airway managements, adequate ventilation and emergency surgical interventions are very important. However, traditional airway managements and respiratory support has only limited effect. In our center, using extracorporeal membrane oxygenation (ECMO) as a novel approach to manage patient with central airway obstruction caused by neck and chest tumors has been adopted. We aimed to show the feasibility: using early ECMO to manage difficult airway, provide oxygenation and support surgical procedure for patients with critical airway stenosis caused by neck and chest tumors. We designed a single-center, small sample size retrospective study based on real-world. We identified 3 patients with central airway obstruction caused by neck and chest tumors. ECMO was used to ensure adequate ventilation to emergency surgery. Control group cannot be established. Because traditional manner very likely led to death of such patients. Details of clinical characteristics, ECMO, surgery and survival outcomes were recorded. Acute dyspnea and cyanosis were the most frequent symptoms. All patients (3/3) showed descending arterial partial pressure of oxygen (PaO2). Computed tomography (CT) revealed severe central airway obstruction caused by neck and chest tumors in all cases (3/3). All patients (3/3) had definite difficult airway. All cases (3/3) received ECMO support and emergency surgical procedure. Venovenous ECMO was the common mode for all cases. 3 patients weaned off ECMO successfully without any ECMO-related complications. Mean duration of ECMO was 3 h (range: 1.5-4.5 h). Under ECMO support, difficult airway management and emergency surgical procedure were finished successfully for all cases (3/3). The mean ICU stay was 3.3 days (range: 1-7 days), and the mean general ward stay was 3.3 days (range: 2-4 days). Pathology demonstrated the tumor dignity for 3 patients including 2 malignant cases and 1 benign case. All patients (3/3) were discharged from hospital successfully. We showed that early ECMO initiation was a safe and feasible approach to manage difficult airway for patients with severe central airway obstruction caused by neck and chest tumors. Meanwhile, early ECMO initiation could ensure security for airway surgical procedure.

Responses of digestive metabolism to marine heatwaves in pearl oysters.

Marine heatwaves (MHWs) have increased in intensity and frequency in global oceans, causing deleterious effects on many marine organisms and ecosystems they support. Bivalves are among the most vulnerable taxonomic groups to intensifying MHWs, yet little is known about the underlying mechanisms. Here, we investigated the impact of MHWs on the digestive metabolism of pearl oysters (Pinctada maxima). Two moderate and severe scenarios of MHWs were performed by increasing seawater temperature respectively from 24 °C to 28 °C and 32 °C for 3 days. When subjected to MHWs and with increasing intensity, pearl oysters significantly enhanced their digestive enzymatic activities, such as lipase and amylase. LC-MS-based metabolomics revealed negative responses in the lipid metabolism (e.g., steroid biosynthesis, glycerophospholipid metabolism, and sphingolipid metabolism), the amino acid metabolism (e.g., glutamate, histidine, arginine, and proline), and the B-vitamins metabolism. These findings indicate that the digestive metabolism of marine bivalves can likely succumb to intensifying MHWs events.

Transcriptomic responses reveal impaired physiological performance of the pearl oyster following repeated exposure to marine heatwaves.

Marine heatwaves are predicted to become more intense and frequent in the future, possibly threatening the survival of marine organisms and devastating their communities. While recent evidence reveals the adaptability of marine organisms to heatwaves, substantially overlooked is whether they can also adjust to repeated heatwave exposure, which can occur in nature. By analysing transcriptome, we examined the fitness and recoverability of the pearl oyster (Pinctada maxima) after two consecutive heatwaves (24 °C to 32 °C for 3 days; recovery at 24 °C for 4 days). In the first heatwave, 331 differentially expressed genes (DEGs) were found, such as AGE-RAGE, MAPK, JAK-STAT, FoxO and mTOR. Despite the recovery after the first heatwave, 2511 DEGs related to energy metabolism, body defence, cell proliferation and biomineralization were found, where 1655 of them were downregulated, suggesting a strong negative response to the second heatwave. Our findings imply that some marine organisms can indeed tolerate heatwaves by boosting energy metabolism to support molecular defence, cell proliferation and biomineralization, but this capacity can be overwhelmed by repeated exposure to heatwaves. Since recurrence of heatwaves within a short period of time is predicted to be more prevalent in the future, the functioning of marine ecosystems would be disrupted if marine organisms fail to accommodate repeated extreme thermal stress.

Post-responses of intertidal bivalves to recurrent heatwaves.

Heatwaves are becoming hotter, longer and more frequent, threatening the survival of intertidal bivalves and devastating their ecosystems. Yet, substantially overlooked are heatwave-induced post-responses, which are important to assess cascading consequences. Here, we investigated responses of intertidal bivalves, Ruditapes philippinarum, to recurrent heatwaves. Physiological and gene expression analyses demonstrated that the mantle tissue of R. philippinarum did not sensitively respond to heatwaves, but revealed post-responses under recovery scenarios. Of 20 genes related to essential physiology and fitness, 18 were down-regulated during the 1st recovery period, but following repeated exposure, 13 genes were up-regulated, in line with significantly increased activities of energy-metabolizing enzymes, and antioxidant and nonspecific enzymes. The down-regulation of genes involved in biomineralization, nevertheless, was observed under recovery scenarios, implying the trade-off between essential physiological and fitness-related functions. These findings pave the way for understanding the physiological plasticity of marine bivalves in response to intensifying heatwaves.

Clinical Application of Traditional Chinese Medicine Therapy for Type 2 Diabetes Mellitus: An Evidence Map.

Type 2 diabetes mellitus (T2DM), a common disease with a complex etiology in the world, is an important risk factor for severe cardiovascular and cerebrovascular diseases. However, treatments of T2DM are mainly based on Western medicine, whose severe side effects make traditional Chinese medicine (TCM) therapy more appealing to patients and clinicians. The overall clinical evidence for different TCM therapies in the treatment of T2DM is still unclear. This study aimed to adopt the evidence-mapping method and integrate the evidence from various researches on this topic, to depict the whole picture of TCM therapies for T2DM. This review included searches of PubMed, Embase, Web of Science, and three major Chinese literature databases (CNKI, VIP, and Wanfang) from inception to November 18, 2021. Two independent reviewers screened the literature, extracted information, and evaluated the quality of all included studies. A systematic review was subsequently performed. In total, 47 studies were reviewed, of which 46 studies (97.9%) were from China and 1 (2.1%) was from Canada. The evidence map was conducted according to different TCM therapies, including herbs or herbal extracts, compounds, powders, decoctions, pills, external treatment, basic theories and treatment principles of TCM, proprietary Chinese medicines, and unspecified TCM integrated therapies. According to the AMSTAR-2 scoring results, 4 papers were rated as high quality, 11 were low quality, and 32 were very low quality. Outcome indicators mainly focused on FBG, HbA1c, 2-h PBG, TC, TG, LDL-C, etc. The results showed that different types of TCM treatment had different improvement effects on the outcome indicators of T2DM. More consistent benefits were observed in the improvement of FBG, HbA1c, and 2-h PBG with treatment regimens based on basic theories and treatment principles of TCM, decoctions and pills, and unspecified TCM integrated therapies. Among herbs, ginger and Coptis root showed more improvement in all outcomes. Compounds, powders, and external treatment showed relatively consistent beneficial effects on the improvement of FBG. No serious adverse events were reported. Overall, the current evidence map provided an intuitive overview of the beneficial effects of TCM therapies in the treatment of type 2 diabetes. This study can be used as a reference for the clinical application of traditional Chinese medicine in T2DM, but due to the low-quality level of the included studies, it should be treated with caution in clinical practices.

Dairy Cows Experimentally Infected With Bovine Leukemia Virus Showed an Increased Milk Production in Lactation Numbers 3-4: A 4-Year Longitudinal Study.

Bovine leukemia virus (BLV) is widespread in global cattle populations, but the effects of its infection on milk quantity and quality have not been clearly elucidated in animal models. In this study, 30 healthy first-lactation cows were selected from ≈2,988 cows in a BLV-free farm with the same criteria of parity, age, lactation number, as well as milk yield, SCS, and composition (fat, protein, and lactose). Subsequently, these cows were randomly assigned to the intervention (n = 15) or control (n = 15) group, and reared in different cowsheds. Cows in the intervention group were inoculated with 1 × phosphate-buffered solution (PBS) resuspended in peripheral blood mononuclear cells (PBMC) from a BLV-positive cow, while the controls were inoculated with the inactivated PBMC from the same individual. From June 2016 to July 2021, milk weight (kg) was automatically recorded by milk sensors, and milk SCS and composition were originated from monthly performed dairy herd improvement (DHI) testing. Fluorescence resonance energy transfer (FRET)-qPCR and ELISA showed that cows in the intervention group were successfully infected with BLV, while cows in the control group were free of BLV for the entire period. At 45 days post-inoculation (DPI), the numbers of whole blood cells (WBCs) (P = 0.010), lymphocytes (LYMs) (P = 0.002), and monocytes (MNCs) (P = 0.001) and the expression levels of IFN-γ (P = 0.013), IL-10 (P = 0.031), and IL-12p70 (P = 0.008) increased significantly in the BLV infected cows compared to the non-infected. In lactation numbers 2-4, the intervention group had significantly higher overall milk yield (P < 0.001), fat (P = 0.031), and protein (P = 0.050) than the control group, while milk SCS (P = 0.038) and lactose (P = 0.036) decreased significantly. Further analysis indicated that BLV infection was associated with increased milk yield at each lactation stage in lactation numbers 3-4 (P = 0.021 or P < 0.001), but not with SCS and milk composition. Together, this 4-year longitudinal study revealed that artificial inoculation of BLV increased the milk yield in cows in this BLV challenge model.

Dissemination of Acupuncture-Moxibustion Clinical Practice Guidelines among Clinical Practitioners: A Systematic Review of Quality Assessment Studies.

Acupuncture clinical practice guidelines are authoritative medical recommendations developed by evaluating and integrating acupuncture-related evidence. However, their synthesis and dissemination are not integrated, and clinical practitioners require more credible effective evidence. The study aim was to systematically review problems disseminating acupuncture clinical practice guidelines to clinical practitioners, to facilitate evidence dissemination. This systematic review included searches of PubMed, EBSCO, Web of Science, and four major Chinese electronic databases (CNKI, VIP, Wanfang Database, and SinoMed) from inception to October 26, 2021. Two independent reviewers screened the literature, extracted information, and evaluated the quality of included studies. A systematic review was subsequently performed. Eleven studies were reviewed: nine (81.8%) cross-sectional surveys and two (18.2%) systematic reviews. The evaluated clinical practice guidelines differed across studies; seven studies (63.6%) evaluated guidelines for a specific disease, one (9.1%) evaluated guidelines for acupuncture therapies (e.g., moxibustion and fire acupuncture), one (9.1%) evaluated US acupuncture guidelines and recommendations, and two (18.2%) did not describe the guideline content. The included studies used different evaluation indicators. Guideline dissemination problems included lack of guideline standardization, unclear target population, mismatch between guidelines and application environment, lack of reliable health economics evaluation, poor quality content of the recommendations, lack of linkage between recommendations and evidence, and disassociation of recommendations from clinical practice et al. The development and publishing of credible acupuncture clinical practice guidelines is urgently needed to improve the usability of guidelines and standardize and disseminate tools for analysing information to clinical practitioners and to help the domestic and international acupuncture community to apply evidence to practice. Recommendations for promoting the dissemination of acupuncture clinical practice guidelines are to define clinical events suitable for the target population, to develop recommendations relevant to clinical practice, to improve the evidence evaluation index system, and to further standardize the method and process of formulating guidelines.

The association between platelet glycoprotein-specific antibodies and response to short-term high-dose dexamethasone with prednisone maintenance treatment in adult patients with primary immune thrombocytopenia.

OBJECTIVE: The aim of the present study was to detect the association between platelet glycoprotein-specific autoantibodies and the patient response to short-term high-dose dexamethasone (HD-DXM) + prednisone maintenance treatment. METHODS: The data from 112 adult patients newly diagnosed with ITP who were administered first-line HD-DXM + prednisone maintenance therapy between January 2016 and January 2021 were retrospectively analyzed. RESULTS: A total of 72 patients positive for platelet glycoprotein-specific antibodies were enrolled in the antibody-positive group, and 40 patients not positive for platelet glycoprotein-specific antibodies were enrolled in the antibody-negative group. In the antibody-positive group, six platelet glycoprotein-specific antibody types were found: 41.67% of the patients were anti-GP IIb/IIIa-positive only, 5.56% were anti-GP Ib/IX-positive only, 5.56% were anti-P-selectin-positive only, 19.44% were anti-GP IIb/IIIa- and anti-GP Ib/IX-positive, 16.67% were anti-GP Ib/IX- and P-selectin-positive and 11.11% were positive for all three antibodies. There was no significant difference in the overall response rate between the antibody-positive group and the antibody-negative group (94.44 versus 80.00%, p = .221). However, the CR rate was significantly higher in the antibody-positive group than in the antibody-negative group (69.44% versus 40.00%, p = .032). The logistic regression analysis revealed that platelet glycoprotein-specific antibody positivity and age were two factors that could affect patient response. CONCLUSIONS: The present study discovered that adult patients newly diagnosed with ITP who had positive platelet glycoprotein-specific antibody test results were likely to achieve a better response after treatment with HD-DXM + prednisone maintenance.

Elemental Sulfur-Promoted Benzoxazole/Benzothiazole Formation Using a C═C Double Bond as a One-Carbon Donator.

An efficient method to assemble diverse benzoxazoles/benzothiazoles in good yields was developed via oxidative cyclization with 2-aminothiophenols or 2-iodoanilines as raw materials. In this protocol, elemental sulfur was used as the effective oxidant and C atoms on the C═C double bond were introduced as a one-carbon donator.

Prevalence and molecular characterization of Wolbachia in field-collected Aedes albopictus, Anopheles sinensis, Armigeres subalbatus, Culex pipiens and Cx. tritaeniorhynchus in China.

Wolbachia are maternally transmitted intracellular bacteria that can naturally and artificially infect arthropods and nematodes. Recently, they were applied to control the spread of mosquito-borne pathogens by causing cytoplasmic incompatibility (CI) between germ cells of females and males. The ability of Wolbachia to induce CI is based on the prevalence and polymorphism of Wolbachia in natural populations of mosquitoes. In this study, we screened the natural infection level and diversity of Wolbachia in field-collected mosquitoes from 25 provinces of China based on partial sequence of Wolbachia surface protein (wsp) gene and multilocus sequence typing (MLST). Among the samples, 2489 mosquitoes were captured from 24 provinces between July and September, 2014 and the remaining 1025 mosquitoes were collected month-by-month in Yangzhou, Jiangsu province between September 2013 and August 2014. Our results showed that the presence of Wolbachia was observed in mosquitoes of Aedes albopictus (97.1%, 331/341), Armigeres subalbatus (95.8%, 481/502), Culex pipiens (87.0%, 1525/1752), Cx. tritaeniorhynchus (17.1%, 14/82), but not Anopheles sinensis (n = 88). Phylogenetic analysis indicated that high polymorphism of wsp and MLST loci was observed in Ae. albopictus mosquitoes, while no or low polymorphisms were in Ar. subalbatus and Cx. pipiens mosquitoes. A total of 12 unique mutations of deduced amino acid were identified in the wsp sequences obtained in this study, including four mutations in Wolbachia supergroup A and eight mutations in supergroup B. This study revealed the prevalence and polymorphism of Wolbachia in mosquitoes in large-scale regions of China and will provide some useful information when performing Wolbachia-based mosquito biocontrol strategies in China.

Integrative Analysis of miRNA and mRNA Expression Profiles in Mammary Glands of Holstein Cows Artificially Infected with Staphylococcus aureus.

Staphylococcus aureus- induced mastitis is one of the most intractable problems for the dairy industry, which causes loss of milk yield and early slaughter of cows worldwide. Few studies have used a comprehensive approach based on the integrative analysis of miRNA and mRNA expression profiles to explore molecular mechanism in bovine mastitis caused by S. aureus. In this study, S. aureus (A1, B1 and C1) and sterile phosphate buffered saline (PBS) (A2, B2 and C2) were introduced to different udder quarters of three individual cows, and transcriptome sequencing and microarrays were utilized to detected miRNA and gene expression in mammary glands from the challenged and control groups. A total of 77 differentially expressed microRNAs (DE miRNAs) and 1625 differentially expressed genes (DEGs) were identified. Gene Ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that multiple DEGs were enriched in significant terms and pathways associated with immunity and inflammation. Integrative analysis between DE miRNAs and DEGs proved that miR-664b, miR-23b-3p, miR-331-5p, miR-19b and miR-2431-3p were potential factors regulating the expression levels of CD14 Molecule (CD14), G protein subunit gamma 2 (GNG2), interleukin 17A (IL17A), collagen type IV alpha 1 chain (COL4A1), microtubule associated protein RP/EB family member 2 (MAPRE2), member of RAS oncogene family (RAP1B), LDOC1 regulator of NFKB signaling (LDOC1), low-density lipoprotein receptor (LDLR) and S100 calcium binding protein A9 (S100A9) in bovine mastitis caused by S. aureus. These findings could enhance the understanding of the underlying immune response in bovine mammary glands against S. aureus infection and provide a useful foundation for future application of the miRNA-mRNA-based genetic regulatory network in the breeding cows resistant to S. aureus.

Isolation and characterization of multidrug-resistant Klebsiella pneumoniae from raw cow milk in Jiangsu and Shandong provinces, China.

Antimicrobials are the most important therapy to bovine mastitis. Bacterial infection and antibiotic treatment of mastitis cycles frequently in dairy farms worldwide, giving rise to concerns about the emergence of multidrug-resistant (MDR) bacteria. In this study, we examined the microbial diversity and antibiotic resistance profiles of bacteria isolated from raw milk from dairy farms in Jiangsu and Shandong provinces, China. Raw milk samples were collected from 857 dairy cattle including 800 apparently healthy individuals and 57 cows with clinical mastitis (CM) and subjected to microbiological culture, antimicrobial susceptibility assay and detection of antibiotic-resistant genes by polymerase chain reaction (PCR) and sequencing. A total of 1,063 isolates belonging to 41 different bacterial genera and 86 species were isolated and identified, of which Pseudomonas spp. (256/1,063, 24.08%), Staphylococcus. spp. (136/1,063, 12.79%), Escherichia coli (116/1,063, 10.91%), Klebsiella spp. (104/1,063, 9.78%) and Bacillus spp. (84/1,063, 7.90%) were most frequently isolated. K. pneumoniae, one of the most prevalent bacteria, was more frequently isolated from the farms in Jiangsu (65/830, 7.83%) than Shandong (1/233, 0.43%) province, and showed a positive association with CM (p < .001). The antimicrobial susceptibility assay revealed that four of the K. pneumoniae isolates (4/66, 6.06%) were MDR bacteria (acquired resistance to ≥three classes of antimicrobials). Furthermore, among 66 isolates of K. pneumoniae, 21.21% (14/66), 13.64% (9/66) and 12.12% (8/66) were resistant to tetracycline, chloramphenicol and aminoglycosides, respectively. However, all K. pneumoniae isolates were sensitive to monobactams and carbapenems. The detection of antibiotic-resistant genes confirmed that the ß-lactamase genes (blaSHV and blaCTX-M ), aminoglycoside modifying enzyme genes [aac(6')-Ib, aph(3')-I and ant(3″)-I], tetracycline efflux pump (tetA) and transposon genetic marker (intI1) were positive in MDR isolates. This study indicated that MDR K. pneumoniae isolates emerged in dairy farms in Jiangsu province and could be a potential threat to food safety and public health.

Isolation and expression analysis of two novel C-repeat binding factor (CBF) genes involved in plant growth and abiotic stress response in mangrove Kandelia obovata.

Kandelia obovata is one of the cold tolerant mangrove plants along the China coast. To reveal the cold tolerant mechanism of K. obovata, the present work isolated two CBF/DREB1 genes (designated KoCBF1 and KoCBF3) from cold-stressed K. obovata and characterized their expression profiles in various organs and in response to multiple abiotic stresses. The deduced proteins of KoCBF1 and 3 all contain specific features of CBFs, and show high similarity to AmCBF1 and 3 from Avicennia marina, respectively. Different expression patterns of the two CBF orthologous under various abiotic stresses and exogenous hormone suggested that they may have different regulators and be involved in different regulatory pathway. The high basal and cold induced expression of the two genes indicated that they may all play important roles in growth and cold resistance of plants. The significant induction of KoCBF3 after salt and lead (Pb2+) treatments suggested that this CBF gene may also participate in response to salinity and heavy metal stresses. This study will provide a better understanding of CBF-regulated stress-resistant mechanism, which may be benefit in mangrove biotechnological breeding, high-latitude transplanting, and bioremediation of heavy metal pollutions.

Isolation and expression analysis of a CBF transcriptional factor gene from the mangrove Bruguiera gymnorrhiza.

The present work isolated a CBF/DREB1 gene from mangrove Bruguiera gymnorrhiza (BgCBF1) and compared its expression levels in various tissues under normal condition and cold stress, and in leaves exposed to various environmental stimuli. Results showed that the BgCBF1 deduced protein showed almost 100% similarities to that of AcCBF1 from Aegiceras corniculatum and AmCBF1 from Avicennia marina. Real-time quantitative PCR analysis showed that BgCBF1 gene displayed constitute expression in leaf, stem and root samples of plantlets under normal condition, but with different expression levels and tissue preference. When exposed to cold, BgCBF1 could be rapidly, slightly and transiently induced in all tissues. Furthermore, the BgCBF1 gene in leaves displayed a transient and small induction after salt and drought (PEG) exposure, while exhibited relatively high up-regulated expression after the phytohormone abscisic acid (ABA) treatment. These results suggest that the BgCBF1 gene may participate in the ABA mediated development and protection of plant against cold and drought. Further studies on its promoters and downstream genes will be needed to better understand its functions.

Molecular characterization of bovine leukemia virus reveals existence of genotype 4 in Chinese dairy cattle.

Bovine leukemia virus (BLV) causes enzootic bovine leucosis and is widely spread worldwide, except several European countries, Australia and New Zealand. Although BLV is highly prevalent in China, information about the genetic diversity and evolutionary dynamics of BLV among Chinese dairy herds is still lacking. To determine the genetic variability of BLV, 219 cows from four cities of Ningxia province of China were screened for BLV infection by fluorescence resonance energy transfer (FRET)-PCR and sequencing, 16 selected positive samples were subjected to molecular characterization. Phylogenetic analysis using the neighbor-joining (NJ) method on complete sequences of envelope (env) gene of BLV obtained from China and those available in GenBank (representing BLV genotypes 1-10) revealed that those Chinese strains belonged to genotypes 4 and 6. Totally, 23 mutations were identified and 16 of them were determined to be unique mutations among Chinese strains. Alignment of the deduced amino acid sequences demonstrated six mutations in glycoprotein 51 (gp51) and three mutations in glycoprotein 30 (gp30) located in the identified neutralizing domain (ND), CD8+ T cell epitope, E-epitope, B-epitope, gp51N12 and cytoplasmic domain of transmembrane protein. This study reported for the first time the BLV genotype 4 in China, and further studies are warranted to compare its immunogenicity and pathogenicity with other BLV genotypes.