To obtain human antibodies against the Gn protein of Severe fever with thrombocytopenia syndrome virus (SFTSV) with phage display technology, this study aimed to screen anti-Gn protein antibodies from an anti-SFTSV Fab human phage display library. Antibody genes were identified by sequence analysis and the specificity of antibodies was confirmed by ELISA. The Fab antibody genes were cloned into the HL51-14 vector and expressed in a mammalian cell expression system. IgG antibodies were then purified by protein A affinity chromatography,and the results were further confirmed by ELISA,IFA,western blotting assays and micro-neutralization tests. The results showed that, after three rounds of panning, there were 390 human Fab antibodies against SFTSV particles, of which 364 were specific for nucleoprotein. Coated with the Gn protein, eight different Fab antibodies specific for Gn protein were obtained after the determination of the subtype and subclass of antibodies by gene sequencing; five of these antibodies were from the Lambda library and three were from the Kappa library. The eight IgG antibodies could specifically bind to Gn protein according to the ELISA, IFA and Western blotting assays. The micro-neutralization test showed that these eight antibodies had no neutralizing activity,but they could still provide a reference for research in human monoclonal antibodies against SFTSV.
Antibodies ; genetics ; immunology ; Bunyaviridae Infections ; genetics ; immunology ; virology ; Cell Line ; Cloning, Molecular ; Humans ; Immunoglobulin Fab Fragments ; genetics ; immunology ; Immunoglobulin G ; genetics ; immunology ; Neutralization Tests ; Phlebovirus ; genetics ; immunology ; Viral Proteins ; genetics ; immunology

The polymorphism in the exon 4 of FcRL3 gene was evaluated by PCR-FPLR in 506 patients with Graves' disease (GD), 80 with Hashimoto thyroiditis (HT) and 261 normal subjects in Chongqing. The data suggest that 82G allele in exon 4 of FcRL3 gene may be susceptible to GD in male patients of Chongqing Hart nationality.

Serum cortisol levels during oral glucose tolerance test (OGTY) were measured in subjects of type 2 diabetic pedigrees. The results showed that cortisol levels during OGTF were higher in type 2 diabetic patients than those in non-diabetic first-degree relatives and normal controls. Fasting cortisol level was positively correlated with fasting plasma glucose level in type 2 diabetic pedigree members. These results suggest that the dysregulation of hypothalamic-pituitary-adrenal axis may coexist in type 2 diabetic patients.

Objective To investigate the change of serum non-esterified fatty acid (NEFA) level in nondiabetic first-degree relatives of type 2 diabetics, and to explore the related factors in the change.MethodsSerum lipid profile, plasma glucose and insulin levels were measured in 186 type 2 diabetic patients, 565 nondiabetic first-degree relatives of type 2 diabetics and 149 normal controls. Results (1) The fasting NEFA level in first-degree relatives was significantly lower than that of type 2 diabetic patients [(0.53±0.28 vs 0.63±0.31) mmol/L,P＜0.01]and HOMA-IR was significantly higher than that of normal controls (0.98±0.51 vs 0.89±0.47,P＜0.01). (2) The fasting NEFA level in the first-degree relatives with higher body mass index (BMI), plasma glucose or area under curve of glucose concentration (AUCglu) was higher than that in those with lower BMI, plasma glucose , blood pressure or AUCglu (all P＜0.05). (3) NEFA showed significantly positive correlations with BMI, systolic blood pressure, diastolic blood pressure (DBP), AUCglu in the first-degree relatives by correlative analysis (r=0.12, r=0.148, r=0.21 and r=0.281 respectively, all P＜0.05). Stepwise linear regression analysis showed that DBP, AUCglu and age were the independent risk factors of NEFA (all P＜0.01). Conclusion Insulin resistance exists in nondiabetic first-degree relatives of type 2 diabetics, which seems to be related to elevated NEFA levels.

Objective To explore the variation and influential factors of high sensitive C-reactive protein (hs-CRP) level in type 2 diabetic family members. Methods A total of 427 type 2 diabetic patients, 377 non-diabetic first-degree relatives of type 2 diabetics and 135 normal control subjects without diabetic family history were recruited. Serum hs-CRP, clinical and biochemical parameters were measured. The relations among indicators were analyzed. Results Compared with normal control subjects, serum hs-CRP levels in type 2 diabetics and first-degree relatives were significantly increased (both P＜0.05), and the increment was even marked in type 2 diabetics than that in first-degree relatives (P＜0.01). The serum hs-CRP levels in type 2 diabetics and first-degree relatives were positively associated with body mass index, waist-hip ratio, abdominal circumference, postgrandial 2 h plasma glucose, fasting and postgrandial 2 h serum insulin, HOMA-IR, triglyceride, creatinine and negatively correlated with high density lipoprotein-cholesterol. In first-degree relatives, serum hs-CRP level was positively associated with systolic blood pressure and HOMA-β. Conclusion As in type 2 diabetic patients, there exists inflammatory reaction in the non-diabetic first-degree relatives of type 2 diabetics, which may play an important role in the pathogenesis of type 2 diabetes mellitus.

Objective :To construct hepatocarcinoma specific IL-1? anti-sense RNA expression vector and to explore its effect on the growth of implanted hepatocarcinoma H22 cells in mice and the possible mechanism. Methods:Murine IL-1? anti-sense RNA expression vectors pafpIRES2-antiIL-1?1 and pafpIRES2-antiIL-1?2 under the regulation of minimal alpha-feto protein (AFP) promoter and CMV enhancer were constructed,and further verified by PCR,restriction endonuclease analysis and DNA sequencing. H22 cells transfected with pafpIRES2-antiIL-1? 1 or pafpIRES2-antiIL-1? 2 were divided into 3 groups:H22/mock,H22/antiIL-1?1 and H22/antiIL-1?2 group. Expression of IL-1? was detected by RT-PCR. Transfected H22 cells were subcutaneously injected into mice to establish tumor implanted mouse model. Tumor volume was measured; the cytotocixity of spleen NK against H22 cells was detected by MTT. Results:Hepatocarcinoma specific IL-1? anti-sense RNA expression vectors pafpIRES2-antiIL-1?1 and pafpIRES2-antiIL-1?2 were successfully constructed and were verified by PCR,restriction endonuclease analysis and DNA sequencing. IL-1? expression in H22 cells was down-regulated after transfected with IL-1? anti-sense RNA expression vectors,especially with the pafpIRES2-antiIL-1?2 vector. Hepatocarcinoma cells implanted mouse model was successfully established. Tumor volume and growth of tumor in H22/antiIL-1?2 mice was obviously smaller than that in H22/mock mice,and the cytotocixity of spleen NK against H22 cells in H22/antiIL-1?1 and H22/antiIL-1?2 mice was also greatly enhanced. Conclusion:Hepatocarcinoma specific IL-1? anti-sense RNA expression vector pafpIRES2-antiIL-1? was successfully constructed. It effectively inhibits the growth of implanted hepatocarcinoma in mice probably through specifically blocking expression of IL-1? and increasing cytotocixity of spleen NK.

OBJECTIVE: To explore the characteristics of traditional Chinese medical syndrome (TCM syndrome) of hepatocirrhosis. METHODS: Clinical information from the four diagnosis methods of traditional Chinese medicine (TCM) and related laboratorial indexes were systematically collected from 223 hepatocirrhosis cases, and the multi-statistical methods including systematic cluster analysis, principal component analysis, stepwise discrimination and variance analysis were made with the software SAS 6.11. RESULTS: Multi-analysis showed that there were 3 categories of syndrome characteristics. Type 1 (134 cases): damp heat, blood stasis, deficiency of liver and spleen Qi; Type 2 (62 cases): deficiency of both Qi and Yin with severe deficiency of Qi, heat with severe dampness, blood stasis; Type 3 (27 cases): deficiency of both Qi and Yin with severe deficiency of Yin, stasis and heat or dampness. Analysis of the changes of the related laboratorial indexes among the three types of syndrome showed that Type 1 mainly manifested asthenia syndrome with sthenia syndrome, and its indexes of AST, ALT, GGT levels were markedly higher than those of Type 2 and Type 3, both of which mainly showed sthenia syndrome with asthenia syndrome, and that Type 3 was in active inflammation, deficiency of both Qi and Yin (deficiency of Yin > deficiency of Qi), and its FN, Alb, FV, FVII, PLT, PCT levels were obviously reduced. CONCLUSION: The multi-statistical methods can reveal the characteristics and regularity of TCM syndrome of hepatocirrhosis, and the 3 categories of syndrome characteristics basically conform to clinical manifestations. The result of TCM syndrome distribution and laboratorial indexes infer that damp heat is the pathological basis of hepatocirrhosis, and the degree of liver function disorder and liver damage may be the pathological basis of deficiency of Yin of both liver and kidney.

Aged ; Female ; Humans ; Male ; Middle Aged ; Neoplasms ; genetics ; mortality ; Pedigree

Objective To study the association of transcription factor 7-like 2(TCF7L2)polymorphisms with tvpe 2 diabetes mellitus in Chinese Han population. Methods Two polymorphisms (rs7903146 and rs12255372)of TCF7L2 gene were genotyped in 446 patients with type 2 diabetes mellitus(T2DM group)and 303 normal subiects (NC group) by PCR-restriction fragment length polymorphism(PCR-RFLP).Waist circumference.body mass index,plasma glucose,serum insulin,lipid profiles,high-sensitivity C-reactive protein and non-esterified fatty acid were measured.Homeostasis model assessment of insulin resistance(HOMA-IR)and β-cell function(HOMA-β)were calculated.Results (1) In T2DM group,T allele frequency and CT,TY geno tvpe frequeneies of rs7903146 were significantly higher than those in NC group(0.093,0.150,0.018 vs 0.043, 0.079,0.003,respectively,a11 P

Objective To explore the effect of scenario simulation teaching method in the training of cardiopulmonary resuscitation (CPR) for junior surgical residents. Methods 133 junior residents (working life<3 years) in the author's hospital rotating Cardiac Surgery department were selected and divided into control group (n=65) and observation group (n=68), in which the control group adopted traditional classroom teaching: teacher explanation-demonstration-student practice-teacher counseling; the observation group adopted scenario simulation teaching: teacher explanation-demonstration-student practice-teacher counseling scenario simulation. Before and after training, two groups both received the CPR theory and double operation assessment , comparing the difference of assessment scores between groups before and after training . Results There was no statistically significant difference in the demographic characteristics between the two groups. The assessment scores of CPR theory and operation were improved in both groups after training, and there were significant differences (P=0.000). There were no statistically significant differences in the CPR theory and operation assessment scores between the two groups before training (P>0.05);compared with the scores of CPR operation between the two groups after training, the observation group [(84.62±3.94)] was significantly higher than that of the control group [(79.68±5.45)] and there were significant statistical differences (P=0.000), while there was no statistically significant difference in CPR theory assessment scores between the two groups (P>0.05). Conclusions Both traditional classroom teaching and scenario simulation teaching methods could improve the CPR theory and skill level of the surgical junior resident, but situa tional simulation teaching method is better than traditional classroom teaching method in improving students' CPR performance.