To investigate the antimicrobial resistance of Enterococci isolated from 596 anal swabs,feces and environmental samples were collected from pig,cattle,chicken and quail farms in Chang-sha,Hunan Province,Enterococci were isolated and identified by mass spectrometry.The minimum inhibitory concentrations(MICs)of 10 antimicrobial agents were determined by AGAR diffusion method.Whole genome sequencing(WGS)was used to detect the distribution of multilocus se-quence typing(ST),drug resistance genes and virulence genes.A total of 272 strains of Enterococ-cus were isolated(45.6％).The isolates were resistant to cefoxitin(68.9％)and cefotiofur(58.5％),followed by trimethoprim/sulfamethoxazole(52.2％),vancomycin(4.4％),and linezolid(13.6％).In this study,six linezolid highly resistant Enterococci isolates were analyzed by whole genome sequencing to explore the transmission mechanism of linezolid resistance because linezolid is forbidden to be used in aquaculture.ST403(4/6),ST16(1/6)and ST476(1/6)were the most common ST types,which all originated from the same farm.Three oxazolidinone resistance genes(cfr,poxtA and optrA)were found in all 21 strains.One strain(Ecc60)carried all three oxazo-lidinone resistance genes,but none of them were located on the plasmid.tet(M),aph(3')-Ⅲ,and lsa(A)were found in all six Enterococcus isolates.Interestingly,the present study is the first to i-dentify enterococci carrying the optrA gene in quail fecal samples.The analysis of the genetic envi-ronment of oxazolidinone resistance genes showed that the genetic environment of cfr(D),poxtA and optrA of isolates from the same farm was similar.A total of 19 virulence genes were detected in 6 isolates,of which 12 genes(ElrA,SrtA,ace,agg,cCF10,cOB1,cad,camE,ebpA,ebpC,efaAfs,tpx)were carried by all 6 isolates,and the types of virulence genes in strains from the same farm were extremely similar.The results showed that the drug resistance of Enterococci iso-lated from the fish farms in Changsha was serious,and the resistance rate to linezolid,which was prohibited in the fish farms,was high.The oxazolidinone resistance gene loci were accompanied by other resistance genes,especially the florfenicol resistance gene(FexA),which may be related to the abuse of florfenicol in the fish farms.

ObjectiveTo analyze the epidemiological characteristics of influenza virus in severe acute respiratory tract infection (SARI) cases in Jingzhou City, so as to provide a scientific basis for the formulation of influenza prevention and control policies in Jingzhou City. MethodsSARI surveillance was carried out in two sentinel hospitals in Jingzhou City from 2018 to 2023. Respiratory tract samples were collected from cases and influenza virus nucleic acid was measured using real-time fluorescent polymerase chain reaction (RT-PCR). ResultsA total of 2 603 SARI samples were tested from 2018 to 2023, and 338 samples were positive for influenza virus nucleic acid, with a detection rate of 12.99%. The highest positive detection rate was 20.22% in 2019, followed by 14.29% in 2022, and the lowest detection rate was 7.75% in 2020. There were significant differences for the positive detection rates of influenza in each monitoring year (χ²=30.386, P<0.001). There were epidemic peaks in the five surveillance years from 2018 to 2023 except 2020. There were winter epidemic peaks during 2018‒2019 and 2021‒2022, and an obvious summer epidemic peak was also observed from 2019 to 2022. H1N1, H3N2, B-Victoria and B-Yamagata were alternately prevalent in the six surveillance years. In 2019, H1N1, H3N2 and B-Victoria were alternately prevalent with time progress, in 2021 only B-Victoria was prevalent, and in 2022 H3N2 and B-Victoria were prevalent. There was no statistically significant difference for the positive detection rates of influenza virus between different genders (χ²=0.178, P=0.673). Among the four age groups, the positive rate of influenza virus in the age group of 15‒<25 years old was the highest (40.91%), followed by the age group of 25‒<60 years old (21.31%). There were statistically significant differences for the positive rates of influenza virus among different age groups (χ²=24.496, P<0.001). ConclusionThe surveillance of SARI cases in Jingzhou City could serve as an effective supplement to the surveillance of ILI in sentinel hospitals. It is suggested to expand the surveillance scope, strengthen public education and outreach on the prevention and control of respiratory diseases, thereby providing a scientific basis for influenza prevention and control.

To investigate the antimicrobial resistance of Enterococci isolated from 596 anal swabs,feces and environmental samples were collected from pig,cattle,chicken and quail farms in Chang-sha,Hunan Province,Enterococci were isolated and identified by mass spectrometry.The minimum inhibitory concentrations(MICs)of 10 antimicrobial agents were determined by AGAR diffusion method.Whole genome sequencing(WGS)was used to detect the distribution of multilocus se-quence typing(ST),drug resistance genes and virulence genes.A total of 272 strains of Enterococ-cus were isolated(45.6％).The isolates were resistant to cefoxitin(68.9％)and cefotiofur(58.5％),followed by trimethoprim/sulfamethoxazole(52.2％),vancomycin(4.4％),and linezolid(13.6％).In this study,six linezolid highly resistant Enterococci isolates were analyzed by whole genome sequencing to explore the transmission mechanism of linezolid resistance because linezolid is forbidden to be used in aquaculture.ST403(4/6),ST16(1/6)and ST476(1/6)were the most common ST types,which all originated from the same farm.Three oxazolidinone resistance genes(cfr,poxtA and optrA)were found in all 21 strains.One strain(Ecc60)carried all three oxazo-lidinone resistance genes,but none of them were located on the plasmid.tet(M),aph(3')-Ⅲ,and lsa(A)were found in all six Enterococcus isolates.Interestingly,the present study is the first to i-dentify enterococci carrying the optrA gene in quail fecal samples.The analysis of the genetic envi-ronment of oxazolidinone resistance genes showed that the genetic environment of cfr(D),poxtA and optrA of isolates from the same farm was similar.A total of 19 virulence genes were detected in 6 isolates,of which 12 genes(ElrA,SrtA,ace,agg,cCF10,cOB1,cad,camE,ebpA,ebpC,efaAfs,tpx)were carried by all 6 isolates,and the types of virulence genes in strains from the same farm were extremely similar.The results showed that the drug resistance of Enterococci iso-lated from the fish farms in Changsha was serious,and the resistance rate to linezolid,which was prohibited in the fish farms,was high.The oxazolidinone resistance gene loci were accompanied by other resistance genes,especially the florfenicol resistance gene(FexA),which may be related to the abuse of florfenicol in the fish farms.

Objective To determine the blood physiological and biochemical indexes in the inbred SHANXI MU strain of spontaneous type 2 diabetes(T2DM)Chinese hamsters.Methods Chinese hamsters with spontaneously developed T2DM and normal hamsters(n=10 hamsters per group),aged 12 months,were selected for the study.Fasting blood samples were collected and 15 physiological parameters and 16 biochemical indicators were analyzed using a Sysmex XT automated hematology analyzer and Hitachi fully automatic biochemical analyzer.Results The white cell count,red cell count,platelet count,hemoglobin level,alanine transaminase,aspartate transaminase,glutamate,total cholesterol,triglycerides,and uric acid all differed significantly between the diabetic and control groups(P＜0.05).Conclusions The change of blood physiological and biochemical indexes in the Chinese hamster spontaneous T2DM model were in line with the trend in human T2DM incidence,thus providing basic data for the application of this model.

Objective To investigate the mechanism of Chaihuang Qingyi Huoxue Granules(Bupleuri Radix,unprocessed Rhei Radix et Rhizoma,Paeoniae Radix Rubra,Paeoniae Radix Alba,Cortex Magnoliae Officinalis,etc.)on rats with severe acute pancreatitis(SAP)based on the renin-Ang-Ⅰ otensin system(RAS).Methods Sixty-four SD rats were randomly divided into four groups:sham operation group,model group,Chaihuang Qingyi Huoxue Granules group(4.42 g·kg-1)and Captopril group(5 mg·kg-1).Each group was further divided into 12-hour and 24-hour subgroups,with 8 rats in each group.SAP rat model was replicated by retrograde injection of 3.5％sodium taurocholate into the biliopancreatic duct.The Captopril group was intraperitoneally injected with Captopril(5 mg·kg-1),and the Chaihuang Qingyi Huoxue Granules group was given intragastric administration,once every 6 hours.The serum amylase(AMY)activity was detected by biochemical method at 12 hours and 24 hours after operation.The pathological changes of pancreatic tissue were observed by HE staining.Serum aldosterone(ALD)content was detected by chemiluminescence.Serum Renin,angiotensin converting enzyme(ACE)and angiotensin Ⅱ(Ang-Ⅱ)were detected by ELISA.The expression of AT1R protein in pancreatic tissue was detected by Western Blot.Results In the same subgroup at 12 and 24 hours,compared with the sham operation group,the serum AMY activity of rats in the model group was significantly increased(P<0.05),the pathological score of pancreatic tissue was significantly increased(P<0.05),the levels of serum ALD,Renin,Ang-Ⅱ and ACE were significantly increased(P<0.05),and the expression of AT1R protein in pancreatic tissue was significantly up-regulated(P<0.05).Compared with the model group,the serum AMY activity of rats in Chaihuang Qingyi Huoxue Granules group and Captopril group was significantly decreased(P<0.05),the pathological score of pancreatic tissue was significantly decreased(P<0.05),the levels of serum ALD,Renin,Ang-Ⅱ and ACE were significantly decreased(P<0.05),and the expression of AT1R protein in pancreatic tissue was significantly down-regulated(P<0.05).Compared with the Captopril group,the serum AMY of the rats in the Chaihuang Qingyi Huoxue Granules group was significantly decreased(P<0.05),the pathological score of pancreatic tissue was significantly decreased(P<0.05),and the serum ALD,Renin,Ang-Ⅱ and ACE levels were significantly decreased(P<0.05).Conclusion Chaihuang Qingyi Huoxue Granules may inhibit the production of Renin and ALD by down-regulating the expression of ACE-Ang-Ⅱ-AT1R classical axis,thus exerting a protective effect on SAP rats.

Gastroesophageal reflux disease （GERD） is a frequently and commonly occurring disease in clinic. In recent decades， with the development in pathophysiology and drug researches， modern medicine has achieved remarkable progress and results in diagnosis and treatment. However， the treatments for non-erosive reflux disease， refractory gastroesophageal reflux disease， proton pump inhibitor resistance， overlap of disease symptoms， and extraesophageal symptoms are limited and ineffective. Traditional Chinese medicine （TCM） was widely used in clinical practice， which has been proved effective in relieving symptoms and improving the quality of life. Sponsored by China Association of Chinese Medicine （CACM） and undertaken by the Spleen and Stomach Disease Branch of CACM， "the 12^th Youth Salon of Clinical Predominance Disease Series （GERD）" invited 18 authoritative digestive experts of TCM and western medicine to discuss "the difficulties of clinical diagnosis and treatment of GERD and TCM advantages". The focus issues such as modern medical diagnosis and treatment achievements and contributions， improvement and maintenance of symptoms， response to overlapping disease symptoms， reduction and withdrawal of acid suppressors， and treatment of extra-esophageal symptoms were discussed in depth. TCM and western medicine exchanged and complemented each other's strengths， combing the difficulties of modern medical diagnosis and treatment， which clarified the positioning and advantages of TCM and provided guidance for clinical and scientific research.

In response to the market demand for therapeutic antibodies, the upstream cell culture scale and expression titer of antibodies have been significantly improved, while the production efficiency of downstream purification process is relatively fall behind, and the downstream processing capacity has become a bottleneck limiting antibody production throughput. Using monoclonal antibody mab-X as experimental material, we optimized the caprylic acid (CA) precipitation process conditions of cell culture fluid and low pH virus inactivation pool, and studied two applications of using CA treatment to remove aggregates and to inactivate virus. Based on the lab scale study, we carried out a 500 L scale-up study, where CA was added to the low pH virus inactivation pool for precipitation, and the product quality and yield before and after precipitation were detected and compared. We found that CA precipitation significantly reduced HCP residuals and aggregates both before and after protein A affinity chromatography. In the aggregate spike study, CA precipitation removed about 15% of the aggregates. A virus reduction study showed complete clearance of a model retrovirus during CA precipitation of protein A purified antibody. In the scale-up study, the depth filtration harvesting, affinity chromatography, low pH virus inactivation, CA precipitation and depth filtration, and cation exchange chromatography successively carried out. The mixing time and stirring speed in the CA precipitation process significantly affected the CA precipitation effect. After CA precipitation, the HCP residue in the low pH virus inactivation solution decreased 895 times. After precipitation, the product purity and HCP residual meet the quality criteria of monoclonal antibodies. CA precipitation can reduce the chromatography step in the conventional purification process. In conclusion, CA precipitation in the downstream process can simplify the conventional purification process, fully meet the purification quality criterion of mab-X, and improve production efficiency and reduce production costs. The results of this study may promote the application of CA precipitation in the purification of monoclonal antibodies, and provide a reference for solving the bottleneck of the current purification process.
Cricetinae ; Animals ; Antibodies, Monoclonal/metabolism* ; Caprylates/chemistry* ; Cell Culture Techniques ; Chromatography, Affinity ; CHO Cells ; Cricetulus ; Chemical Precipitation

Micronutrients are important factors affecting the immune system. Micronutrients contribute to maintaining the normal functions of immune system through maintaining the integrity of skin and mucosa, regulating the activity of cytokines, and mediating the proliferation and differentiation of T cells and B cells. Human immune system changes at different stages of growth and development, and micronutrient deficiency can impair the immune system and increase the susceptibility to respiratory infections. Therefore, maintaining the optimal nutritional status is beneficial for the immune system to prevent respiratory infections. Severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2) infection is one of the respiratory infectious diseases that is of most concern recently. Micronutrient supplementation has been shown to have positive effects on the prevention of, symptom relief of and recovery from SARS-Cov-2 infection. Further studies are needed to systematically evaluate the effects of micronutrients on immunity and respiratory infections, and to identify the appropriate timing and dosage.

Objective:To evaluate nutritional status and to analyse risk factors of acute exacerbation of chronic obstructive pulmonary disease (COPD) in the elderly.Methods:Data of elderly hospitalized patients with COPD mainly from 5 grade A, class 3 hospitals (Beijing Hospital, Shanghai Huadong Hospital, Tianjin Nankai hospital, the Second Affiliated Hospital of Medical College of Zhejiang University, Guangzhou First People′s Hospital) from January 2012 to December 2020 were retrospectively analyzed. According to the global initiative for chronic obstructive lung disease criteria (GOLD), elderly COPD patients were divided into acute exacerbation group and stable group. The differences in age, gender, medical history, anthropometry, laboratory examination, nutritional support, results of nutritional risk screening 2002 (NRS2002) and global leadership initiative on malnutrition (GLIM) were compared between the two groups after admission, and the risk factors of acute exacerbation of COPD in the elderly were analyzed by multivariate Logistic regression.Results:A total of 339 elderly patients with COPD aged 65-100 years were included in this study, including 177 cases (52.21%) in acute exacerbation stage. The detection rate of malnutrition in acute exacerbation stage was higher than that in stable stage (51.98% vs 41.98%, P<0.05). The weight, body mass index and grip strength of patients in the acute exacerbation stage were significantly lower than those in the stable period [(55.47±8.42) vs (60.63±9.30) kg, (20.52±4.25) vs (22.39±4.57) kg/m 2, (12.32±4.21) vs (16.59±2.97) kg] (all P<0.05). Spearman correlation analysis showed that the acute exacerbation of elderly patients with COPD was positively correlated with malnutrition ( r=0.443, P<0.001), and negatively correlated with body weight, body mass index and calf circumference ( r=-0.200, -0.214, -0.135, all P<0.05). Multiple Logistic regression analysis showed that acute exacerbation in elderly patients with COPD was only related to malnutrition ( OR=7.799, 95% CI: 4.466-13.622, P<0.001). Conclusions:The incidence of malnutrition in acute exacerbation stage of elderly COPD patients is high. Malnutrition is independently related to acute exacerbation of COPD.