Objective To express the capsid proteins VP1 and VP3 of hepatitis A virus(HAV) in prokaryotic cells and evaluate their immunogenicity.Methods VP1 and VP3 gene fragments were amplified by PCR,cloned into vector pETG28a to construct recombinant expression plasmids pET-G28a-VP1 and pET-G28a-VP3,which were transformed into competent E.coli BL21(DE3),induced by IPTG,and then analyzed by 12% SDS-PAGE.The target protein was purified by ion exchange chromatography,renatured and combined with several adjuvants to immunize mice.The mice were divided into VP1-MF59,VP1-AH,VP1-AP,VP1-AP-10CpG,VP1-AP-50CpG,VP3-AP,VP3-VP1-AP and PBS control groups,five for each group.Serum IgG antibody titers of mice in various groups were detected by ELISA,and serum neutralizing antibody titers of mice in VP1-AP,VP3-VP1-AP and PBS control groups were detected by rapid fluorescence focus immunosuppression experiment.Results Colony PCR and sequencing showed that the recombinant plasmids pET-G28a-VP1 and pET-G28a-VP3were constructed correctly.The recombinant proteins VP1 and VP3,with relative molecular masses of about 37 000 and26 000 respectively,mainly existed in the form of inclusion bodies,the expression levels were 18.6% and 32.4%,and the purity was 86.3% and 84.7%,respectively.The recombinant proteins VP1 and VP3 reacted specifically with rabbit anti-HAV antiserum and mouse anti-HAV-VP3 antiserum respectively.The serum IgG antibody titer of mice in VP1-AP-50CpG group was significantly higher than that in VP1-AP group(q=22.05,P <0.01),and the serum IgG antibody titer of mice in VP3-VP1-AP group was significantly higher than that in VP1-AP group and VP3-AP group(q=22.05 and 22.49 respectively,each P <0.01).Compared with PBS control group,the serum neutralizing antibody titer of mice in VP1-AP and VP3-VP1-AP group increased significantly(q=7.79 and 25.11 respectively,P<0.01) Conclusion Prokaryotic HAV capsid proteins VP1 and VP3 showed high purity and good immunogenicity,and the addition of CpG in the preparation was beneficial to enhance the immunogenicity of antigens.This study laid a foundation of the development of HAV recombinant subunit vaccine.

Objective To explore the most suitable equation in accessing renal function for the elderly type 2 diabetic patients, and its clinical utility in combination with hypersensitive C-reactive protein (hsCRP). Methods The new Cystatin C-based equations for estimated glomerular filtration rate (Cys-eGFR) and conventional predictive equations were compared with isotopic GFR (iGFR) by linear regression analysis, paired t-test, Bland and Altman procedures and non-parametric receiver operating characteristic (ROC) curves. The new Cys-eGFR equation and hsCRP were also incorporated for detecting renal disease in this population. Results The new Cys-eGFR equation had a better relativity with iGFR (r= 0.767, P＜0.001), a less bias (bias: 0.0007, P＞0.05), a higher conformance (2SD: 21.56), higher sensitivity (90.7%) and specificity (88.6%) for diagnosing moderate decrease in renal function. There was a negative relationship between the new Cys-eGFR and hsCRP (r=-0.655, P＜0.01). When the new Cys-eGFR was 67.06 ml· min-1 ·1.73 m-2 and hsCRP was 5.65 mg/L, the combination of Cys-eGFR and hsCRP was better than the combination of serum creatinine and urine albumin/creatinine ratio in screening stage 3 chronic kidney disease (95%vs.46%). Conclusions The combination of new Cys-eGFR equation and hsCRP may screen an early decrease of moderate GFR.

Smart dressings, which show obvious advantage and potential in wound treatment and real-time monitoring, attract widespread attention in recent years. Real-time and dynamic acquiring wound information is vital to the treatment and prognosis of wound. Further research on smart dressings is helpful for wound management, personalized treatment, and realization of medical application translation of health monitoring technology. In the article, we categorize smart dressings and conclude their functions according to the type of micro-environment information of wound gathered by smart dressings.

Objective To analyze the possible risk factors of IVIG non-response of Kawasaki disease (KD),Shanghai Children's Hospital and Shanghai Junze Software develop an research platform,which is based on E-Science model.Through the mathematical model by integrating the risk factors to explore the method of effective prediction for IVIG non-response,and to provide the clinical basis for timely and effective treatment and prognosis of the disease.Methods The data of KD children who were hospitalized in Shanghai Children's Hospital from January 2013 to November 2016 were included.The indexes included gender,age,time of IVIG treatment,and laboratory examinations.The multivariate logistic regression was used to analyze the influencing factors of IVIG non-response.The indexes in the model were deduced according to the independent variables of the logistic regression equation.The ROC curve and the area under the curve were calculated for the new prediction model.The sensitivity and specificity of the new prediction model were calculated according to the cutoff value.Finally,the new model was compared with the Kobayashi and Egami scoring model.Results The levels of CRP,NLR,LDH,ALB and FDP in children with KD were influencing factors for IVIG nonresponse (P ＜ 0.05).According to the logistic regression equation,the sensitivity and specificity of the model used to predict IVIG non-response were 69.7％ and 80.4％,respectively,and the area under the ROC curve was 0.825 (95％ CI:0.769-0.882).Kobayashi and Egami scoring models were tested,the sensitivity and specificity of the new scoring system were better than previous ones.Condusion The scoring model established in this study has a good effect in predicting IVIG non-response in KD patients and could be used in clinical practice,and it is worthy to be validated and adjusted by large-scale data.

English abstract has become an important part of thesis. As a potential and highly educated professional people,full-time postgraduate nursing students can promote international exchanges and cooperation in nursing. What they write in the full English papers or English abstracts are not only the carrier to enhance mutual understanding between international peers,but also spreading Chinese culture and promoting nursing international influence. Through consulting,guiding and modifying massive literature,we analyze existing problems of nursing graduate thesis' s abstracts and put forward appropriate recommendations,in order to continuously improve the quality of nursing postgraduate thesis and English level,and to increase the academic utilization rate,and make it play its role in the international academic exchanges.

Objective: To investigate the situations of AIDS related knowledge, sexual behavior with various partners among HIV positive patients in Shenzhen before highly active anti-retroviral therapy, and to provide evidences for health education intervention. Methods: Questionnaire survey and information collection were carried out among HIV positive patients, under the informed consent, during the first physical examination before highly active anti-retroviral therapy. Results: The overall awareness rate of AIDS knowledge was 91%, 94.46% and 80.95% respectively for homosexual and heterosexual populations, with significant difference between the two groups (χ²=21.254, P<0.001). In addition, there were significant differences in the factors related to education, income, route of infection, marital status, residence and other related factors (P<0.05). In HIV positive MSM, primary sexual behavior occurred mainly at the age of less than 30 (96.8%) and in university or earlier stage (58.3%), college students as the first sexual partner accounted for 44.9%, and once used Rush accounted for 42.7%. There were significant differences in the number of temporary sexual partners and condom use frequency between homosexual and heterosexual subjects (P<0.05). Conclusions Individuals infected with HIV via different routes are variant on AIDS cognition. Health education on AIDS related knowledge and sexual security before and after antiviral treatment can not only improve the awareness rate of HIV/AIDS infection, but also can reduce the risk of the spread of HIV/AIDS.

Objective: To analyze the genetic characteristics of a five generations pedigree with homozygous familial hypercholesterolemia (HoFH). Methods: Prospective study. Twenty family members included a proband diagnosed as familial hyperlipidemia at the cardiology Department of Xi′an Children′s Hospital in October 2018 were research object. Clinical data were collected. Genome DNAs were extracted. Whole exons sequencing was performed on the proband using target capture next generation sequencing. Candidate gene mutation sites identified by bioinformatics were verified by Sanger sequencing in the family members. The genotype-phenotype correlation of the pedigree was analyzed between heterozygous mutation carriers and non-carriers. Results: The proband was a 7-years and 10-month-old boy. He was born with a roundgreen bean size yellow skin protuberance in the skin of the coccyx. Since the age of 3-4 years old, xanthoma-like lesions with a diameter of 0.5-1.5 cm gradually appeared in the skin of bilateral elbow joints, knee joints and Achilles tendon. The height, weight and intellectual development of the child were the same as those of normal children at the same age. No similar xanthoma-like lesion was found in the other family members. The proband′s total cholesterol (TC) reached 18.16-21.24 mmol/L, and his low density lipoproteincholesterol (LDL-C) was 14.08-15.51 mmol/L. Carotid ultrasonography showed diffuse sclerotic plaques in bilateral carotid and vertebral arteries, and color Doppler echocardiography revealed aortic valve thickening and calcification. Gene testing identified that the proband carried a homozygous mutation C. 418G>A (p. E140K) in LDLR gene inherited from his parents who had a consanguineous marriage and carried a heterozygous mutation of LDLR-E140K, respectively.The TC, LDL-C and apolipoproteinB (ApoB) of LDLR-E140K gene heterozygous carriers ((8.40±0.13), (6.79±0.01) and (1.95±0.05) mmol/L, respectively) were significantly higher than those of non-carriers ((4.59±0.28), (3.35±0.39) and (0.86±0.10) mmol/L, t=7.269, 4.595, 6.311, respectively, P<0.05). Conclusions LDLR-E140K gene homozygous mutation is first reported to be associated with most severe phenotype HoFH. The genotype-phenotype analysis of the pedigree shows that the clinical phenotype of the proband with homozygous mutation is the most serious, and all the heterozygous mutation carriers present with hypercholesterolemia phenotype. The investigation confirms that LDLR-E140K is the pathogenic variation of familial hyperlipidemia.