Objective:To construct a recombinant gene of single-chain fragment variable(scFv)against the SARS-CoV-2 spike protein(S protein).Methods:Using genetic engineering antibody technology,BALB/c mice were immunized with SARS-CoV-2 S protein antigen.The total RNA of the spleen was extracted to obtain the heavy chain(VH)and light chain(VL)gene sequences of the antibody variable region(V).Through multiple-primer PCR amplification and screening,the corresponding VH and VL gene sequences were obtained.After purification and recovery,the VH and VL primer genes of anti-SARS-CoV-2 S protein with restriction enzyme cutting sites and the VH and VL genes were successfully connected by the overlap extension PCR method to obtain the scFv recombinant gene.The sequencing results were compared and analyzed in the gene bank by using BLAST on the NCBI website.Results:The VH and VL genes of anti-SARS-CoV-2 S protein were successfully obtained,and scFv was obtained by connecting through the overlap extension PCR technique(SOE-PCR).The BLSAT analysis of scFv showed that its gene homology was greater than 80%.The indirect ELISA detection results showed that there was obvious binding in the prokaryotic expression supernatant.Conclusion:This study provides a technical basis for the diagnosis and treatment of SARS-CoV-2 and the research and development of new related antibodies.

Objective:To construct a recombinant gene of single-chain fragment variable(scFv)against the SARS-CoV-2 spike protein(S protein).Methods:Using genetic engineering antibody technology,BALB/c mice were immunized with SARS-CoV-2 S protein antigen.The total RNA of the spleen was extracted to obtain the heavy chain(VH)and light chain(VL)gene sequences of the antibody variable region(V).Through multiple-primer PCR amplification and screening,the corresponding VH and VL gene sequences were obtained.After purification and recovery,the VH and VL primer genes of anti-SARS-CoV-2 S protein with restriction enzyme cutting sites and the VH and VL genes were successfully connected by the overlap extension PCR method to obtain the scFv recombinant gene.The sequencing results were compared and analyzed in the gene bank by using BLAST on the NCBI website.Results:The VH and VL genes of anti-SARS-CoV-2 S protein were successfully obtained,and scFv was obtained by connecting through the overlap extension PCR technique(SOE-PCR).The BLSAT analysis of scFv showed that its gene homology was greater than 80%.The indirect ELISA detection results showed that there was obvious binding in the prokaryotic expression supernatant.Conclusion:This study provides a technical basis for the diagnosis and treatment of SARS-CoV-2 and the research and development of new related antibodies.

Background/Aims: Exacerbating factors of ulcerative colitis (UC) are multiple and complex with individual influence. We aimed to evaluate the efficacy of disease control by searching and restricting inflammation trigger factors of UC relapse individually in daily clinical practice. Methods: Both patients with UC history or new diagnosis were asked to avoid dairy products at first doctor visit. Individual-reported potential trigger factors were restricted when UC flared up (Mayo endoscopy score ≥1) from remission status. The remission rate, duration to remission and medication were analyzed between the groups of factor restriction complete, incomplete and unknown. Results: The total remission rate was 91.7% of 108 patients with complete restriction of dairy product. The duration to remission of UC history group was significantly longer than that of new diagnosis group (88.5 days vs. 43.4 days, P=0.006) in patients with initial endoscopic score 2–3, but no difference in patients with score 1. After first remission, the inflammation trigger factors in 161 relapse episodes of 72 patients were multiple and personal. Milk/dairy products, herb medicine/Chinese tonic food and dietary supplement were the common factors, followed by psychological issues, non-dietary factors (smoking cessation, cosmetic products) and discontinuation of medication by patients themselves. Factor unknown accounted for 14.1% of patients. The benefits of factor complete restriction included shorter duration to remission (P<0.001), less steroid and biological agent use (P=0.022) when compared to incomplete restriction or factor unknown group. Conclusions Restriction of dairy diet first then searching and restricting trigger factors personally if UC relapse can improve the disease control and downgrade the medication usage of UC patients in daily clinical practice.

BACKGROUND/OBJECTIVES: Heavy alcohol consumption causes the development of alcoholic liver disease (ALD), a neglected but important public health problem. Many studies have pointed out that probiotics could improve gut health, which is also considered to be a cause of ALD. Therefore, this study screened the probiotics, Lactobacillus casei GKC1 (GKC1), L. fermentum GKF3 (GKF3), Bifidobacterium lactis GKK2 (GKK2), L. rhamnosus GKLC1 (GKLC1), L. paracasei GKS6 (GKS6), and L. plantarum GKM3 (GKM3), for their potential benefits in alleviating ALD for applications to disease prevention. SUBJECTS/METHODS: C57BL/6N mice were divided into 8 groups (n = 6 in each): normal control, positive control (alcohol-diet fed), and treatments of feeding probiotics GKC1, GKF3, GKK2, GKLC1, GKS6, and GKM3 under an oral dose 0.82 g/kg B.W. per day by oral gavage. The experiment was conducted for 8 weeks, and the concentrations of alanine aminotransferase (ALT), aspartate aminotransferase, triglyceride (TG), and total cholesterol (TC) in mice were measured. The glutathione (GSH), catalase (CAT), and histology were analyzed after sacrifice. RESULTS: The results showed a decrease in the serum ALT, liver TG, and liver TC levels in the GKS6, GKM3, and GKLC1 groups compared to the positive control. In addition, the decreasing GSH and CAT levels were inhibited in the GKS6 and GKM3 groups. The histopathological results showed that all probiotics could reduce the accumulation of liver fat. Furthermore, there was a significant difference in GKLC1 with lower stomach damage compared to the alcohol-fed mice without any addition of probiotics. CONCLUSIONS GKLC1, GKS6, and GKM3 can be used as supplements for alleviating the development of ALD.

OBJECTIVE:To provide reference for pharmacists to participate in the management of chronic disease. METHODS:A total of 259 patients with chronic airway disease [included asthma and chronic obstructive pulmonary disease (COPD)] met the inclusion criteria were selected from our hospital and 5 community health care centers of medical consortium. These patients received medication safety assessment management,which was led by clinical pharmacists of our hospital with the participation of community pharmacists,including medication safety comprehensive evaluation and risk classification management, follow-up and medication guidance, integrated prescriptions checking, establishment of shared database. 1 years after the implementation,the effectiveness were evaluated by score the relatived indicators in related groups. RESULTS:After a year of the management mode practice,compared with before intervention,the patients'safety medication cognitive ability score in high-risk and low-risk group increased from(4.49±1.26)and(7.31±1.01)to(5.40±1.56)and(7.44±0.91);medication adherence score increased from(4.96±1.21)and(7.08±1.24)to(6.66±1.08)and(7.38±0.98);ACT score from asthma patients increased from (16.15±2.58)and(21.15±1.03)to(16.80±2.57)and(21.64±1.55);CAT score from COPD patients decreased from(25.51± 4.07) and (14.90 ± 3.95) to (24.20 ± 3.96) and (13.80 ± 4.08);the rate of irrational prescription effective identification and intervention by pharmacists increased from 3.6％ and 1.4％ to 9.4％ and 7.6％,respectively. All the differences above were statistically significant (P<0.05). CONCLUSIONS:The participation of pharmacists in long-term medication safety assessment management for chronic airway disease patients can improve patients'safety medication cognitive ability,medication adherence, disease control and the pharmacists'ability of irrational drug use identification and intervention.