The sfa1 gene encoded a bifunctional enzyme with the activities of both alcohol dehydrogenase and glutathione-dependent formaldehyde dehydrogenase in Saccharomyces cerevisiae.The gene disruption cassette produced by PCR using the same long oligonucleotides which comprise 19 or 22 nucleotides complementary to sequences in the templates(pUG6 and pUG66 marker plasmid)at 3' end and 45 nucleotides at 5' end that annealed to sites upstream or downstream of the genomic target sequence to be deleted.After two linear disruption cassettes with a Cre/loxP mediated marker were transformed into the cells of Saccharomyces cerevisiae YS-1,the positive transformants were checked by PCR to correct the integration of the cassette and concurrent deletion of the chromosomal target sequence.Once correctly integrated into the genome,the select marker can be efficiently rescued by transformating the plasmid pSH47 into YS-1 and inducing the Cre expression with a Cre/loxP-mediated marker removal procedure.The expression of the Cre recombinase finally resulted in the removal of the marker gene,leaving behind a single loxP site at the chromosomal locus.The diploid mutant YS-1-sfa1 was generated,which could enhance the output of ethanol with 8.0% by shaking culture in flask compared with the original strain YS-1.

Objective: To learn HIV related stigma and its associated factors among the patients on antiretroviral therapy ( ART ) in Wenshan Prefecture, Yunnan Province, so as to provide evidence for eliminating HIV discrimination. Methods: A total of 419 subjects were recruited by convenience sampling from three ART clinics in Wenshan City and Maguan County between October 2017 and January 2018. HIV/AIDS Related Stigma and Discrimination Scale developed by Li Xianhong et al was employed. The multivariate linear regression model were used to explore the influencing factors for HIV stigma. Results: The median scores of disclosure concern, public rejection, family stigma, internalized stigma, health service providers＇ stigma were 24.00, 6.00, 10.00, 20.00, 2.00, respectively, and the overall was 68.00. The multivariate linear regression analysis showed that female patients ( standardized β=0.135 ) , patients with opportunistic infection ( standardized β=0.120 ), patients had no HIV infected family member ( standardized β=-0.128 ) , patients without family support ( standardized β=-0.175 ) , patients received gift from ART clinics ( standardized β=0.124 ) , patients scored lower in ART knowledge ( standardized β=-0.117 ) were likely to scored higher in HIV stigma. Conclusions The stigma on disclosure concern and internalized stigma dimensions are grievous among ART patients in Wenshan Prefecture. Gender, opportunistic infection, HIV infection in family, family support, receiving incentive gifts from clinics and awareness of ART are associated with HIV stigma.

OBJECTIVETo investigate the expression levels of CD40, sCD40L, hs-CRP, WBC in acute coronary syndrome (ACS) patients and the association between CD40-1C/T single nucleotide polymorphism and risk of ACS in Han Chinese, moreover, the regulatory effects of IFN-gamma and fluvastatin on the expression of CD40 in peripheral blood mononuclear cell (PBMNC) were also observed.

METHODS(1) 160 ACS patients and 92 control patients diagnosed by coronary angiography were recruited. Enzyme linked immunosorbent assay, particle enhanced immunoturbidimetric assay, flow cytometry were used to detect the levels of soluble CD40L, hs-CRP, and WBC count. (2) The CD40 genotype and allele frequency were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing technology. (3) PBMNC were separated by density gradient centrifugation heparinized venous blood from 40 ACS patients, cultured for 24 h with or without 100 ng/ml IFN-gamma in the absence or present of 10 micromol/L fluvastatin. The CD40 expression levels were then detected by flow cytometry.

RESULTSInflammatory cytokine CD40, sCD40L, hs-CRP levels were significantly higher in ACS patients than in controls. The CD40-1C allele frequency was 0.606 in ACS group and 0.489 in controls, while T allele frequency was 0.394 in ACS group and 0.511 in controls. The frequency of CC genotype was significantly higher in ACS group than in controls (P < 0.01). C allele carriers had significantly higher risk of ACS (OR = 1.608, 95%CI: 1.12 - 2.32, P = 0.011). CD40 production increased after 24 h culturing and the CD40 levels were significantly higher in subjects with CC genotype than that in subjects with CT or TT genotype [CC: (14.78 +/- 4.56) MFI, CT: (11.98 +/- 4.12) MFI, TT: (9.86 +/- 3.83) MFI, P < 0.05]. IFN-gamma further increased PBMNC CD40 expressions in all subjects after culturing for 24 h and fluvastatin equally inhibited IFN-gamma induced PBMNC CD40 expression from various genotypes (CC, CT, TT was 30.3%, 26.3%, 29.3% respectively, all P > 0.05).

CONCLUSIONInflammatory cytokines were increased in ACS patients and CD40-1C/T polymorphism is associated with higher risk for ACS in Han Chinese.

Acute Coronary Syndrome ; genetics ; Aged ; CD40 Antigens ; genetics ; metabolism ; CD40 Ligand ; genetics ; metabolism ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Humans ; Male ; Middle Aged ; Polymorphism, Single Nucleotide

Aim To investigate the protective effect of Citrus aurantium L. polysaccharides-B(CALB) on ra-diation induced by 60Co γ-ray in mice. Methods The BALB/c mice were randomly divided into blank control group, radiation model group, CALB administration group (high, medium and low dose), and positive control group(black fungus polysaccharide,HP). The mice were administered orally for 30 days. After the last administration for three hours,the survival rates on the 2nd day and the 14th day of the blank control group and the irradiated mice after the single radioac-tive irradiation (7 Gy) with 60Co γ-ray were meas-ured. In addition, DNA content and micronucleus of bone marrow cells, SOD, GSH-Px activities, MDA content in serum, liver and brain tissues in mice, TChE activity in brain tissues and spleen and thymus index of mice were detected after one-time whole body irradiation with 60Co γ-ray (3 Gy). Results Each dose group of CALB could significantly improve the survival rate of irradiated mice,increase the DNA con-tent of mouse bone marrow cells and reduce the number of micronuclei in bone marrow cells. In addition, CALB could also increase the thymus and spleen index and the levels of SOD and GSH-Px in serum,brain and liver tissues of mice,and reduce the content of MDA. Conclusion CALB has protective effect on radiation injury,which can be used for further development and utilization of Fructus aurantii.

Based on variation of Pinus massoniana families, heritablility and correlation analysis, the contents of shikimic acid and procyanidine (heritability 0.90, 0.70), dry weight of single branch (heritability 0.60) and and leaf length (heritability 0.46) were screened out as quality, yield and harvest cost traits of Folium Pini, respectively. For the different medicinal application of Folium Pini, varied methods were chosen to estimate weight and construct index equation. Weight adjustment based.on equal emphasis were used as economic weight determining method to select the best families, and the index (accuracy 0. 936 4 and heritability 0. 881 6) obtained was a little better than that obtained by equal emphasis, and much better than that by restricted index. The superior families selected with adjustment weight and equal emphasis were No. 46, 43 and 28. Partial regression were used as economic weight determining method to select the best families,and the index obtained had the highest accuracy (0.941 5) , index heritability (0. 889 9) and the genetic gain of shikimic acid content. The superior families selected with this method were No. 46, 27 and 47. No. 46 was the best families with maximal economic benefit. Our study indicated that suitable method for estimate weight and construct index equation can be applied for better accuracy of superior families selection of P. massoniana.
Breeding ; Drugs, Chinese Herbal ; analysis ; Pinus ; chemistry ; classification ; genetics ; growth & development ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; classification ; genetics ; growth & development

BACKGROUND: Procalcitonin (PCT), C-reactive protein (CRP), and white blood cells (WBCs) are inflammatory markers used to diagnose severe bacterial infections. We evaluated the diagnostic role of these markers and compared their accuracy for spontaneous bacterial peritonitis (SBP) associated with chronic severe hepatitis B (CSHB). METHODS: PCT and CRP concentrations, WBC count, and other hematological parameters were measured in serum from 84 well-characterized patients with CSHB, of whom 42 had SBP. Receiver operating characteristics (ROC) curve analysis was performed to assess the diagnostic accuracy. RESULTS: PCT and CRP concentrations were significantly higher in the CSHB patients with SBP (n=42) than CSHB patients without SBP (n=42). PCT and CRP concentrations were more accurate than WBC count for the diagnosis of CSHB-associated SBP. The optimal cutoff value of PCT was 0.48 ng/mL. The PCT concentration was significantly correlated with the CRP concentration and WBC count. CONCLUSIONS: Serum PCT and CRP seems to be better markers than WBC for the diagnosis of CSHB patients with SBP.
Age Factors ; Area Under Curve ; Bacterial Infections/complications/*diagnosis ; Biological Markers/blood ; C-Reactive Protein/*analysis ; Calcitonin/*blood ; Female ; Hepatitis B, Chronic/*complications ; Humans ; Leukocyte Count ; Leukocytes/cytology ; Male ; Middle Aged ; Peritonitis/complications/*diagnosis ; Protein Precursors/*blood ; ROC Curve ; Sex Factors ; Temperature

The purpose of this investigation is to improve ethanol production and decrease acetate formation in Saccharomyces cerevisiae strain YS2-?adh2.The strain YS2-?adh2 with deleted alcohol dehydrogenase Ⅱ(adh2) gene was isolated in our lab with higher ethanol production than that of the strain YS2.The ace-taldehyde dehydrogenase Ⅵ(ald6) gene encoded a cytosolic acetaldehyde dehydrogenase,a key enzyme of the pyruvate dehydrogenase(PDH) bypass,transfers acetaldehyde to acetate.To disrupt ald6 gene of the strain YS2-?adh2,ald6 gene targeting cassettes were synthesized by long flanking homology PCR(LFH-PCR) and then were transformed into YS2-?adh2 mutants by LiAc/SS Carrier DNA/PEG method.Positive transformants were selected with G418 and further confirmed by PCR.Once correctly integrated into the genome,the selective marker was rescued by transforming the plasmid pSH65 into the positive transformants and inducing the Cre expression with a Cre/loxP-mediated marker removal procedure.We named the ald6 gene knocked-out strain as YS2-?adh2-?ald6 which has a 12.5% higher ethanol production and a 18% lower acetate formation compared to the strain YS2.

Adult ; HIV Seropositivity ; psychology ; Homosexuality, Male ; psychology ; Humans ; Male ; Sexual Behavior

Sino-foreign cooperative education not only introduces foreign teachers and curriculums, but also brings new teaching concepts, teaching methods, and the training of teachers. With the example of rehabilitation treatment in the national first-class undergraduate specialty construction site of Nanjing University of Chinese Medicine, this study discusses the talent training mode of Sino-foreign cooperative education from the five aspects of professional system, discipline characteristics, curriculum system, teaching methods, and evaluation methods. The mode of Sino-foreign cooperative education provides a reference for the development of modern rehabilitation higher education.

OBJECTIVETo investigate the mortality of HIV infected clients from methadone maintenance treatment (MMT) clinics in Yili Kazakh autonomous prefecture as well as the factors associated with mortality of HIV infected clients.

METHODSA retrospective cohort study was performed. Data of 860 cases were collected from National Methadone Maintenance Treatment database, National AIDS/HIV database and antiretroviral therapy (ART) treatment database for adults. Information collected included demographic information of HIV infected clients, methadone daily treatment information, CD4 testing information, ART treatment information and death information. Recruiting began from August, 2005 through May, 2011. Cox proportional regression was used to identify factors associated with mortality. The proportional hazard assumption was assessed using Schoenfeld's residuals test. Missing values were imputed using the multiple linear regression method. R software (version 2.13.0) was used to perform data analysis.

RESULTSA total of 860 HIV positive MMT clients were analyzed. The methadone dose for study subjects was (38.2 ± 20.7) mg/d. 27.8% (239/860) of study subjects participated in ART treatment, 38.7% (333/860) had never tested for CD4 count. The age for study subjects was (32.9 ± 6.4) years old. Among all these subjects, 67.3% (579/860) were married. During the observation period, 151 deaths were observed in 2192.9 person years. The average observation time was 2.6 year for each subject. The all-cause mortality rate was 68.9‰. Cox proportion model showed that ART treatment (HR = 0.53, 95%CI: 0.32 - 0.88), baseline CD4 count at 200 - 350 cells/µl (HR = 0.35, 95%CI: 0.20 - 0.60), baseline CD4 count more than 350 cells/µl (HR = 0.16, 95%CI: 0.09 - 0.29), and marriage (HR = 0.55, 95%CI: 0.37 - 0.82) were associated with less mortality compared with control group. Age (more than 45 years old) (HR = 5.20, 95%CI: 2.60 - 10.20) and sharing needles (HR = 1.40, 95%CI: 1.02 - 2.00) were risk factors associated with death.

CONCLUSIONHigh mortality rate was observed among HIV infected clients. Methadone clinic should provide ART treatment or ART referral services.

Acquired Immunodeficiency Syndrome ; drug therapy ; mortality ; Adolescent ; Adult ; Anti-HIV Agents ; therapeutic use ; China ; Female ; HIV Infections ; drug therapy ; mortality ; Humans ; Male ; Methadone ; therapeutic use ; Middle Aged ; Retrospective Studies ; Survival Rate ; Young Adult