We expressed multi-epitope chimeric protein of CARDS toxin protein of Mycoplasma pneumonia (Mp) in prokaryotic cells,and purified and investigated its immunoreactivity.A recombinant multi-epitope chimeric gene including ten critical epitopes was connected by linker and cloned into prokaryotic expression vector pET-28a(+),and transformed into E.coli BL21(DE3) cells for expression under induction of IPTG.The antigenicity of expressed recombinant protein was identified with 6 × His monoclonal antibody and human positive serum by Western blot.The recombinant expression vector pET-CARDS was constructed and the about 30 kDa recombinant chimeric protein expressed in BL21(DE3) successfully.Western blot analysis showed that it can react respectively with 6 × His monoclonal antibodies and human positive serum.This study showed that the chimeric CARDS protein has an obvious immunoreactivity and a potential to be a new antigen for the diagnosis of Mp infection.

ABSTRACT:To understand the status of contamination and antimicrobial resistance of Staphylococcus aureus (S .aureus) isolated from raw cow milk ,600 samples were collected from five different provinces in China .After isolation and identification of S .aureus from these samples ,157 S .aureus isolates were obtained .Furthermore ,their resistant phenotype and resistant gene were analyzed by MIC detection ,as well as nine drug‐resistance genes determination and analysis .The result revealed that most of the S .aureus isolates contained multiple antibiotic resistant genes ,and all of the isolates were resistant to penicillin . Of 98 .73% isolates were resistant to amoxicillin .Conversely ,all of the isolates were sensitive to enrofloxacin and vancomycin . Moreover ,there were 29 MRSA positive strains among which 22 strains carrying femA gene .In conclusion ,much more atten‐tion should be paid to the drug‐resistance capabilities of field S .aureus isolates in raw cow milk ,and further monitoring of drug resistant S .aureus strains in milk should also be strengthened so as to ensure milk safety .

AIM: To investigate the effect of FAK - related non - kinase ( FRNK) on the expression of membrane - type matrix metalloproteinase -1 ( MT1 - MMP) in hepatic stellate cells ( HSC). METHODS:FRNK were trans-fected into HSCs by cationic liposome method. The protein levels of FRNK in HSC were assayed by Western blotting. The levels of MT1 - MMP were determined by RT - PCR for mRNA and by Western blotting for protein, respectively. RESULTS: The up -regulated expression of FRNK protein was observed and it was at 48 h after transfection that the FRNK protein content was the highest ( P < 0.05 ). The expressions of MT1 - MMP mRNA and protein were also up - regulated by the transfection of FRNK, and it was at 48 h after transfection that the MT1 - MMP protein content was significantly increased. CONCLUSION: The mRNA and protein of FRNK were over - expressed in HSC transfected with the gene of FRNK. The inhibitory effect of FRNK on the collagen synthesis in HSC may be through the up - regulation of MT1 - MMP.

Objective To establish reference values of normal fetal heart conduction time intervals by tissue Doppler imaging(TDI)and pulsed Doppler(PD)echocardiography,and to assess their correlation with gestational-age and fetal heart rate.Methods One hundred and eighty-nine pregnant women underwent detailed echocardiographic examinations.Atrio-ventricular conduction time interval(AV)and the time interval from onset of ventricular contraction to the onset of atrial contraction of next cardiac cycle(VA)were measured by TDI and PD echocardiography.Results TDI-AV was(126.56±15.33)ms(95% CI 124.10～129.03 ms),TDI-VA was(285.22±24.53)ms(95% CI 281.27～289.16 ms),PD-AV was(127.42±12.88)ms(95% CI 125.35～129.49 ms),PD-VA was(287.42±25.19)ms(95% CI 283.37～291.47 ms).A paired t test revealed no systematic difference between the two approaches used to measure AV and VA.AV and VA were significantly positively correlated with gestational age,and significantly negatively correlated with fetal heart rate.Heart conduction time intervals were altered in fetus with paroxysmal arrhythmia.Conclusions This study established the normal values of fetal heart conduction time intervals.Prenatal determination of fetal heart conductional time intervals has important potential clinical utility in assessing fetal arrhythmia.

We investigated serotype and resistance of Salmonella during pig slaughtering in Shandong Province,China,providing basic data for the risk assessment of Salmonella and for guiding the clinical medication.We used rapid classification kit to identify Salmonella serotype,adopted broth microdilution method to detect the resistance of 13 kinds drugs belong to 8 categories.Result showed that the identified 9 kinds of serotype were mainly S.derby and S.typhimurium.The resistance to 13 kinds drugs of 298 Salmonella were different.The higher percentage of tetracycline drugs as Doxycycline(DOX) and Tetracycline(TE) were 97.99％ and 80.20％,respectively,which was most sensitive to Colistine E.The resistant rate was only 2.01 ％,following by Amoxicillin/Clavulanic acid and Ofloxacin which were 2.35 ％ and 4.03％,and the multiple resistant rate was 81.88％.TE-DOX was the regnant drug-resistant spectrum.In conclusion,the predominant serotype of Salmonella in links of pig slaughtering in Shandong Province is S.derby,and resistance is different to the different drugs.The drug resistance of different slaughter links exist significant differences,multiple drug resistance is serious,and drug-resistant spectrum are varied.

Objective:To analyze the level of self-management and its influencing factors in patients with systemic lupus erythematosus (SLE).Methods:A cross-sectional study. From January to December 2021, patients with SLE in the Department of Rheumatology and Immunology in the First Affiliated Hospital of Henan University of Traditional Chinese Medicine were selected as the study objects by convenience sampling method. A total of 135 questionnaires were distributed in this study, 135 were recovered, and 128 were valid. The general data of the patients were collected, and the self-management level, self-efficacy, family care and meaning in life of the patients were evaluated with the chronic disease self-management behavior scale, general self-efficacy scale (GSES), family adaption partnership growth affection resolve index (APGAR) and the meaning in life questionnaire (MLQ). Multiple linear regression analysis was applied to explore the factors affecting the self-management of SLE patients.Results:The total score of chronic disease self-management behavior scale in the 128 SLE patients was (28.77±6.58) points. Pearson correlation analysis showed strong correlation between self-efficacy, sense of life meaning and self-management ( r=0.748, 0.717, both P<0.001), there was a moderate correlation between family care and self-management ( r=0.560, P<0.001). Multiple linear regression analysis showed that educational level ( β′=0.457), disease activity ( β′=-0.211), self-efficacy ( β′=-0.372), family care ( β′=-0.510), meaning in life ( β′=-0.386) were influencing factors of self-management level in patients with SLE (all P<0.05). Conclusions:The self-management level of SLE patients needs to be improved. Disease activity, self-efficacy, meaning in life and other factors can affect the patients′ self-management level.

L-glutamic acid is the world's largest bulk amino acid product that is widely used in the food, pharmaceutical and chemical industries. Using Corynebacterium glutamicum G01 as the starting strain, the fermentation by-product alanine content was firstly reduced by knocking out the gene encoding alanine aminotransferase (alaT), a major by-product related to alanine synthesis. Secondly, since the α-ketoglutarate node carbon flow plays an important role in glutamate synthesis, the ribosome-binding site (RBS) sequence optimization was used to reduce the activity of α-ketoglutarate dehydrogenase and enhance the glutamate anabolic flow. The endogenous conversion of α-ketoglutarate to glutamate was also enhanced by screening different glutamate dehydrogenase. Subsequently, the glutamate transporter was rationally desgined to improve the glutamate efflux capacity. Finally, the fermentation conditions of the strain constructed using the above strategy were optimized in 5 L fermenters by a gradient temperature increase combined with a batch replenishment strategy. The glutamic acid production reached (135.33±4.68) g/L, which was 41.2% higher than that of the original strain (96.53±2.32) g/L. The yield was 55.8%, which was 11.6% higher than that of the original strain (44.2%). The combined strategy improved the titer and the yield of glutamic acid, which provides a reference for the metabolic modification of glutamic acid producing strains.
Glutamic Acid ; Corynebacterium glutamicum/genetics* ; Ketoglutaric Acids ; Metabolic Engineering ; Alanine