Objective To establish a simple and high-performance analytical technique for detecting DNA methylation markers and SNPs simultaneously,and obtain the population genetics data of some SNPs in the hypermethylated region upstream of the human H19 gene.Methods The haplotypes of H19FR1 and H19FR2 which located in the promoter region upstream of the human H19 gene were investigated from 232 unrelated Chinese individuals living in Wuhan by means of PCR and subsequent denaturing gradient gel electrophoresis(DGGE).Based on the methylation status of the genomic DNA,selective detection of the parental alleles for H19FRs was examined by using the methylation-sensitive restriction enzyme(msRE) Hpa II or Hha I.Results Five haplotypes and nine phenotypes were observed for H19FR1 in Chinese Han population in Wuhan,and the power of discrimination(DP),polymorphism information content(PIC) and probability of paternity exclusion(PE) were 0.803,0.58 and 0.322 respectively.For the H19FR2,two haplotypes and three phenotyes were detected,and the DP,PIC and PE were 0.626,0.37 and 0.162 respectively.Sequencing results showed that there were 3 SNPs,a7342g,a7357g and g7547a,and one g7351c point mutation in H19FR1.In the H19FR2,there was only one SNP,a8097g.The msRE,HpaⅡ or HhaⅠ,could digest the maternal allele,and only a single band derived from the paternal allele was detected by post-digestion PCR-DGGE(PDP-DGGE) technique.Conclusion PDP-DGGE is a simple,sensitive and effective technique for analyzing DNA methylation status and SNPs simultaneously,and can be used for discriminating the parental origin of alleles.

Objective To evaluate the therapeutic effectiveness of echo-guided percutaneous radiofrequency ablation(RFA) by using a LeVeen needle electrode combination in the treatment of large liver tumor,and to discuss the relationship between the amount as well as the distribution pattern of the electrodes and the therapeutic effectiveness.Methods During the period of Feb.2006-Dec.2008,echo-guided RFA was performed in 113 patients with large and unresectable liver tumor,with a total of 118 lesions.According to the size of the tumor,the patients were divided into four groups.The tumor’s diameter of Group A(n = 64) was 4.0-5.0 cm,Group B(n = 28) was 5.1-6.0 cm,Group C(n = 11) was 6.1-7.0 cm and Group D(n = 10) was 7.1-9.3 cm.Based on the tumor’s diameter,the amount of the electrodes to be used and the sites to be ablated were determined.After the procedure,the follow-up checks with MRI or CT scanning were carried out to observe the necrotic extent and the local recurrence.Results Two months after the treatment,the complete necrosis rate of the tumor in Group A,B,C and D was 88.4%,78.6%,63.6% and 40.0%,respectively,with an overall necrosis rate of 79.7% in total 118 lesions.During a follow-up period of 3-36 months,the local recurrence rate in Group A,B,C and D was 5.5%,10.0%,28.6% and 50.0%,respectively.Severe complications,including intrahepatic infection(n = 2),puncture passage bleeding(n = 1),symptomatic pleural effusion(n = 4) and biloma(n = 2),occurred in 9 cases.No death related to RFA treatment occurred.Conclusion Echo-guided percutaneous radiofrequency ablation by means of multi-sites superimposition method with a LeVeen needle electrode combination is an safe and effective therapy for the hepatic tumors with the diameter over 4 cm.

Objective To understand the relationship between the requirement and the consumption of the health management, and provide the theoretical basis for the developing of China health management. Methods It took the survey to understand the relationship among 152 medical staffs from the six counties surrounding with Shi Jiazhuang downtown. Results 85.5%of medical staffs always and almost accept health advisory and provide health guidance. There are not the direct relationship between the requirement and the consumption(x2 = 9.39,P＞0.05 ). Health advice and guidance are not translated into the concepts of investment on the health management and disease prevention (x2 = 1.69, P＞0.05 ). The medical staffs don't realize it is important to invest on the health. Conclusions Related industries should strengthen the training on the health management, publicize this profession, promote the development of health management services and improve the national health quality.[ Key words] Health promotion ; Needs assessment; Economics

Safety hospital centralized monitoring is one of the important methods for evaluating the medication safety of TCM injections in clinic.It is critical to warrant research quality by reasonable statistical analysis.However,universally accepted standards and guidelines for the statistical analysis of hospital centralized monitoring of TCM injections have not been issued so far,which would probably reduce the research quality of safety hospital centralized monitoring of TCM injections since the unreasonable use of statistical analysis methods.Combined with previous practical experience and understanding,we put forward the problems of statistical analysis of hospital centralized monitoring of TCM injections with the provision of some advice in this paper based on the analysis of the preceding studies,laying a foundation for the same kind of researches.

ABSTRACT:Objective To investigate the distributional characteristics of anaphylactogens in prurigo nodularis (PN)patients so as to provide clinical basis for selecting the method of allergy test.Methods We divided 262 PN patients into four groups and then used skin-prick (5 4 patients ),skin-patch (5 0 patients ),serum IgE allergen detection (56 patients)and serum IgG allergen detection (102 patients)to analyze anaphylactogen positive rate and their distribution differences in PN.Results For skin-prick test in 54 PN patients,the detection rates of platanus orientalis,artemisia argyi,poplar,dust mite,dirt mite,cod,peanut and tomato were higher than those in the control group (P<0.05 ).For skin-patch test in 50 PN patients,the detection rates of nickel sulfate,flavor compounds,black rubber,and carba mix were higher than those in the control group (P<0.05).For serum IgE test in 5 6 PN patients,the detection rates of house dust,scandent hop,cat/dog hair,bug,penicillium/neurospora and mutton were higher than those in the control group (P<0.05).For serum IgG test in 102 PN ones,the detection rates of crab,shrimp,beef and cod were higher than those in the control group (P<0.05).Conclusion Skin-prick test and serum IgE detection have high application values in PN patients.Serum IgG detection is of certain importance in adjusting PN patients’diet structure while skin-patch test applied in PN needs further exploration.

Objective To investigate the knowledge level of the caregivers about pressure ulcer in patients with high risk of pressure ulcer in rural community,and to provide the scientific basis for the prevention and treatment of pressure ulcer.Methods 216 caregivers for patients with high risk of pressure ulcer in rural community were chosen from Mar.2012 to May 2013.The investigation on the awareness of pressure ulcer was performed by using questionnaire survey.Results The awareness rate of pressure ulcer in caregivers was 41.5 ％.The caregivers had gender difference (174females and 42 males).Most of the caregivers were 41 to 50 years old (124 cases,57.4％).Most caregivers had 1 ～ 2 years work experience (94 cases,43.5％).Most of them had low level of education (170 cases,78.7％) and non-professional training (178 cases,82.4％).The questionnaire score was higher in young caregivers than in the olders(F=2.483,P＜0.05).The questionnaire score was higher in caregivers having a long working lifetime(F=3.624,P＜0.05),higher educationalbackground (F=5.139,P＜ 0.01) and discipline training (t=7.346,P＜ 0.01) than in caregivers not having them.Conclusions The awareness rate of the knowledge about pressure ulcer is low in caregivers for patient with high risk of pressure ulcer in rural community.It is necessary to strengthen the guidance and training in caregivers to reduce the incidence of pressure ulcers and the medical cost,and improve the quality of life in patients with high risk of pressure ulcer in rural community.

Objective To investigate microRNA expression profiles of colonic cancer without lymph node metastasis and identify the specific microRNAs associated with carcinogenesis of colon.Methods Cancerous and para-cancerous tissues (5 cm from cancer tissues of the colon) confirmed without lymph node metastasis were collected from 3 patients. The microRNAs were extracted and isolated by mirVana RNA kit. Hybridizations were made by applying the microRNAs to Agilent microRNA microarray. Data analysis was done by software of Feacture Extraction. The discovered microRNAs were confirmed by real time PCR assay. Results Twelve out of 14 microRNAs associated with colonic cancer were up-regulated in colonic tissues. They were miR-106b, miR-135b,miR-18a,miR-18b,miR-196b, miR-19a, miR-224, miR-335, miR-424, miR-20a' , miR-301b and miR-374a. The rest two of miR-378 and miR 378* were down-regulated. Two up-regulated microRNAs (miR-18a and miR-135b) were detected in 3 pairs of cancerous and para-cancerous tissues. The expression level of miR-18a and miR-135b were accordant with the results of real time PCR.Conclusions microRNA.such as miR-106b,miR-135b.miR-18a.miR-18b,miR-196b,ect,were differentially expressed between cancerous and para-cancerous tissues.Many of these target genes are supposed to participate in the process of multiple tunmorgenesis.These microRNAs play an important role in the carcinogenesis of colon.

Objective:To report two cases of post-transplantation lymphoproliferative disorders (PTLD) after kidney transplantation in children and review the literature, and to improve clinicians' understanding of PTLD in children.Methods:The clinical data of two children with PTLD admitted to the Children's Hospital of Fudan University were collected and analyzed. The PTLD-related literature of PubMed, Embase, Web of Science, Scopus, Cochrane Library, Wanfang, CNKI, Weipu Database and China Biomedical Literature Service System from the establishment of the database to January 2020 were collected for literature review. Multivariate logistic regression analysis method was used to analyze the influencing factors of prognostic in children with PTLD.Results:Both of the patients had negative Epstein-Barr virus (EBV) and Cytomegalovirus (CMV) before transplantation and anti-thymocyte immunoglobulin (ATG) were induced during transplantation. PTLD in case 1 and case 2 was diagnosed at 3 and 12 months after transplantation, respectively, with positive EBV and CMV serological reaction. The pathological diagnosis was monomorphic PTLD in case 1 and the case 2 was clinically considered as non-hodgkin lymphoma. They all received thrapies of immunosuppressive reduction combined with anti-CD20 monoclonal antibody and chemotherapy. PTLD was relieved and graft function was normal in 2 cases, while case 1 died two and half years after transplantation due to intracranial fungal infection. According to the analysis of 56 children (including 2 cases in this study) with PTLD from the literature review, the median time of PTLD from transplantation was 41.8 months. The initial involved organs were digestive tract [17 cases (30.4%)], respiratory system [8 cases (14.3%)], nervous system [7 cases (12.5%)] and pharyngeal lymph ring [7 cases (12.5%)], respectively. The main pathologic type of PTLD was monomorphic [34 cases (60.8%)]. Fifty-six cases were all positive in EBV serological reaction when PTLD was diagnosed. The treatment included immunosuppressive reduction combined with anti-CD20 monoclonal antibody and chemotherapy. Forty-eight cases of PTLD were relieved, while 8 cases lost graft function. Eleven cases died, including 3 cases due to infection and the other 8 cases due to PTLD. Multivariate logistic regression showed that monomorphic PTLD was a risk factor of death for PTLD children ( OR=21.616, 95% CI 1.007-464.107, P=0.049). Conclusions:PTLD in children with kidney transplantation is mostly associated with EBV infection, and the clinical manifestations are diverse. Monomorphic PTLD has a poor prognosis and high mortality.

Objective To assess the effect of silent mating type information regulation 2 homolog 6 (SIRT6) on migratory and proliferative activity of skin fibroblasts in the elderly,and to explore their mechanisms.Methods Circumcised foreskins were obtained from patients of different ages in Department of Urologic Surgery,Second Hospital of Shanxi Medical University,including 8 elderly patients and 8 young patients.Human skin fibroblasts were isolated from the foreskin tissues by using a collagenase digestion method.Western blot analysis was performed to determine the expression of SIRT6 and phosphorylated p65 (p-p65) in human skin fibroblasts in different age groups,scratch assay to evaluate cell migratory activity,and cell counting kit-8 (CCK8) assay to assess cellular proliferative activity.Skin fibroblasts in the elderly group were divided into 2 groups:SIRT6 group transfected with a lentiviral vector overexpressing SIRT6,and control group transfected with an empty lentiviral vector.Then,the cell migratory and proliferative activity as well as p-p65 expression were measured by the above methods,and the mRNA expression of type Ⅰ and Ⅲ collagens,and integrin subunits α3,α5 and β1 was determined by real-time PCR in the SIRT6 group and control group.Statistical analysis was carried out with GraphPad Prism 5 software by using t test for comparison between 2 groups.Results Compared with the young group,the elderly group showed significantly decreased SIRT6 expression in skin fibroblasts (0.434 ±0.179 vs.1.000 ± 0.067,t =3.040,P =0.012),migration rate (43.81％ ± 18.84％ vs.94.63％ ± 12.32％,t =5.903,P =0.003)and cellular proliferative activity at 24 and 48 hours (both P ＜ 0.05),but significantly increased p-p65 expression (1.694 ± 0.148 vs.1.000 ± 0.093,t =2.949,P =0.015).Compared with the control group,the SIRT6 group showed significantly decreased p-p65 expression (P ＜ 0.05),but significantly increased migratory and proliferative activity (both P ＜ 0.05),and elevated mRNA expression of type Ⅲ collagen and integrin subunits oα3,α5 and β1 (all P ＜ 0.05).Conclusion SIRT6 can improve the migratory and proliferative activity of human fibroblasts in the elderly,possibly by inhibiting the nuclear factor-κB pathway.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its variants continue to spread around the world, posing a great threat to global economic and public health. Influenza A virus, a major cause of influenza, has a wide host range and evolves rapidly. Influenza pandemics, which occur every 20 to 40 years, annual seasonal influenza epidemics, severe influenza and animal influenza viruses are all serious threats to public health. Clinical manifestations of infections caused by the two pathogens are very similar. It is of great significance to develop an effective bivalent vaccine for controlling alternating epidemics or co-infection of the two pathogens. This article reviewed the progress in the development of bivalent vaccines against SARS-CoV-2 and influenza A virus based on different platforms.