Objective To study the proteins related to aging in aortic of old rats for laying the foundation of further study of aging mechanism.Methods The rat model of aging was built,and all model rats were divided into 4 groups:the adult group(9 weeks),the old group(12 months)of WistarKyoto (WKY) rats,and the age-matched spontaneously hypertensive rats (SHR).Blood pressure of 4 groups was observed.Morphological change of aorta was observed by HE staining.Differential proteins were identified by isobaric tags for relative and absolute quantitation,(iTRAQ)-coupled liquid chromatography and tandem mass spectrometry technology.Part of differential proteins was subsequently detected by real time PCR and Western blot.Results The mean SBP of the old group SHR was higher than WKY of 97.1％ (t=39.00,P＜0.05),and the adult group of SHR was also higher than WKY of 5.4％(t=3.64,P＜0.05).Compared with the adult group,aging change in the aortic morphology of old SHR and WKY were shown in HE staining,and the change in SHR rats was more marked.7 proteins related to aging were identified by Mass spectrum analysis,and they were Profilin-1,Prelamin A,HSP70,creatine kinase-M,Fibulin-5,eIF5A and Prohibitin.Part of differential proteins was subsequently confirmed by real time PCR and Western blot.Prelamin-A was up-regulated in the old group of WKY and SHR (0.15±0.01 vs.0.45±0.04,0.34±0.02 vs.0.78±0.06) (t=12.67,12.06,all P＜0.01),Prohibitin was down-regulated in the old group of WKY and SHR(1.34±0.05 vs.1.01± 0.06,1.24±0.05 vs.0.88±0.08) (t=7.41,7.09,all P＜0.01).Profilin-1 was up-regulated in the old group of WKY and SHR (9.12±0.4 vs.20.76±0.8,16.84±0.5 vs.55.16±0.9) (t=22.55,64.46,both P＜0.01),and Profilin-1 expression in the old group of SHR was higher thanWKY (55.16±0.90 vs.20.76±0.8,t=49.49,P＜0.01).Conclusions Differential proteins of the old rat aorta are identified through the comparative proteomics method.These differential proteins will provide new targets for the prevention and control of vascular aging.

This article discussed the construction of healing environment in hospitals with reference to the author′s experience in the construction of large hospitals in the last 10 years.This environment requires to improve such aspects as the direct hospital environment identification, privacy, safety,ecology, comfort,artistry and humanism of the hospital.It also depends on improving the indirect environment for the medical workers who apply significant impact on the mentality of patients.These efforts contribute greatly to building a hospital of healing environment.

This paper described the principles and the arrangements on setting-up new clinics in the venues and designated hotels during the G20 Summit of Hangzhou.Personnel allocation,combination of equipment,disposal process and supporting tasks of the clinics were introduced to provide reference for setting similar medically supporting-clinics during the international conference activities.

Objective To construct lentivirus vectors carrying 16 short hairpin RNA (shRNA) expression cassettes targeting histone acetyltransferases and provide a powerful research approach to explore the mechanism of epigenetic genes in regulating hepatitis B virus (HBV).Methods Following the rule of short shRNA primer design,eight-pair primers (A ~ H )for each gene,which had stable interfering efficiency,were designed.The annealed primers were connected to the empty lentiviral vectors of shRNA for transformation.In order to confirm the positive clones,clones were analyzed by real-time polymerase chain reaction (RT-PCR ).Then, qualified plasmids were verified by enzyme digestion technology.Four shRNA lentivirus plasmids against the same gene were mixed to build lentivirus respectively.After the virus transfected into 293T cells for 48 and 72 hours,supernatants were collected to infect HepG2.2.15 cells.The percentage of fluorescent cells were observed and assessed by microscope 72 hours after infection.Results One hundred and twenty-eight lentiviral vectors of RNA interference (RNAi)library were constructed against 16 histone acetyltransferases and more than 80% of HepG2.2.15 cells were infected with lentivirus 72 hours after infection.Conclusions Sixteen shRNA lentivirus vectors against histone acetyltransferase are successfully constructed.Thus,a solid foundation for the study of the effect of human histone deacetylase on HBV replication is established.

Objective: To learn the population,virus status and viral types of hantavirus（HV）hosts in Tiantai County of Zhejiang Province from 2011 to 2018,and to provide evidence for hemorrhagic fever with renal syndrome（HFRS）control. Methods: Rodents in Tiantai County were captured by night trapping method. After the species and age of rodents were identified,the composition of rodent species,dominant species and density of rodents were analyzed. The lungs and blood of rodents were sampled to detect the antigen and antibody of HV by immunofluorescence method. The HV antigen-positive lung samples were detected by RT-PCR with specific primers of HV S fragment,then HV was isolated and identified by inoculating Vero-E6 cells. Results: The average rodent density in Tiantai County from 2011 to 2018 was 4.44%. The rodent density in the field and residential areas were 4.94% and 2.23%,respectively. Ten species of rodents were identified,with Apodemus agrarius dominant in the field and Rattus norvegicus in the residential areas. Sixty-seven lung samples were HV antigen positive（4.13%）,one from Rattus norvegicus and the other sixty-six from Apodemus agrarius. Seventy-nine blood samples were HV antibody positive（4.86%）,all from Apodemus agrarius. Thirty-four HV antigen-positive lung samples were positive（50.75%）after RT-PCR amplification. Twenty-two strains of virus were isolated and all of them were from Apodemus agrarius,including twenty-one strains of Hantaan type（HTN）and one strain of Seoul type（SEO）. Conclusion In Tiantai County,Apodemus agrarius is the main source of HFRS infection;the main epidemic type of HV is HTN and SEO is first found in Apodemus agrarius.

Objective:To analyze the construction and operation of the 19th Hangzhou Asian Games designated hospitals, and to discuss the medical emergency security work of large-scale sports events, so as to provide references for the planning of designated hospitals in future large-scale sports events.Methods:Retrospective analysis was made on the establishment principles, requirements, selection of medical support personnel, and training exercises of the designated hospitals, focusing on the key links such as organizational system, staffing, designated areas, and drug management.Results:Total of 40 designated hospitals have successfully completed the task of medical security by rebuilding the medical security area of the Asian Games, elevating the process, equipping facilities, and strengthening staff training. During the Asian Games, 349 people were transferred to designated hospitals by ambulance, 54 people were hospitalized, 19 people underwent surgery, and 1022 people went to designated hospitals by themselves.Conclusion:The construction of the designated hospitals during the 19th Hangzhou Asian Games was of high quality, efficient and smooth operation. It is suggested that efforts should be made in the reconstruction of the medical security area for the Asian Games to be "relatively independent". The treatment process of self-visiting patients should be fully considered and the flat urgent emergency response mechanism needs to be established.

BACKGROUNDOuabain is a mammalian adrenocortical hormone that is involved in the pathogenesis of hypertension by inhibiting Na-K ATPase activity. It also participates in a variety of kinase-mediated signaling pathways associated with Na-K ATPase. Previous studies have shown that ouabain can cause cardiac remodeling independent of elevated blood pressure and that proliferating cell nuclear antigen (PCNA) plays a coordinating role for numerous proteins involved in multiple processes associated with DNA synthesis. Therefore, we hypothesized that ouabain might play a role in the cerebral cortex through signaling pathways independent of hypertension. And PCNA might be involved in this process.

METHODSMale Sprague-Dawley rats were treated with ouabain or with 0.9% nitric sodium as the control group. Systolic blood pressure was recorded weekly. After four weeks of treatment, morphological changes in the cerebral cortex were analyzed using light and transmission electron microscopy. The expression of PCNA in the cerebral cortex was evaluated by immunohistochemistry, real time quantitative PCR, and Western blotting.

RESULTSAfter 4-week treatment, there was no significant difference in systolic blood pressure compared with the control group, but both structural deterioration and up-regulated expression of PCNA in the brain was induced by ouabain treatment.

CONCLUSIONSThese results suggest that ouabain induces alterations in the brain structure, and this effect is independent of blood pressure. PCNA might be involved in the repair process of ouabain-induced brain damage.

Animals ; Blood Pressure ; drug effects ; Hypertension ; drug therapy ; metabolism ; Immunohistochemistry ; Male ; Ouabain ; therapeutic use ; Proliferating Cell Nuclear Antigen ; metabolism ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Sodium-Potassium-Exchanging ATPase ; metabolism

Objective: By investigating the genotype and evolutionary variation of hantavirus (HV) in Tiantai county, a national surveillance site for hemorrhagic fever with renal syndrome (HFRS) was set in Zhejiang province, from 2011 to 2018, to reveal the molecular epidemiological characteristics of hantavirus (HV) in Tiantai. Methods: Total RNA was extracted from ultrasound treated HV antigen- positive rat lung samples in Tiantai from 2011 to 2018. After cDNA was prepared, nested PCR was used to amplify partial sequence of M fragments by using specific primers of HV. The sequences of HV in Tiantai from 2011 to 2018 were compared with other known HV sequences in order to identify the genotype and analyze the evolution and variation of the virus. Results: In 67 HV antigen-positive lung specimens, 31 were positive in nested PCR amplification with type-specific primers, including 30 Hantaan virus (HTNV) positive samples, 1 Seoul virus (SEOV) positive sample, and all the 31 samples were from Apodemus agrarius. The phylogenetic tree based on partial M segment was divided into monophyletic group, 30 strains were distributed in HTNV group and 1 was in SEOV group. The HTNV strain Tiantai T2018-130 was independently in one branch, sharing 84.8%-87.9% homology with other strains both at home and abroad, including 29 strains in HTNV group in Tiantai. The other 29 HTNV strains in Tiantai showed closer relationship. The SEOV strain T2016-31 from Apodemus agrarius showed closer relationship with previous strains of SEOV, Tiantai ZT71, ZT10 and Z37 strains of Wenzhou, Zhejiang province. Conclusions HTNV, the main genotype of HV in Tiantai of Zhejiang province, showed obvious geographic clustering, but the strain T2018-130 was distinct from the others in Tiantai. Meanwhile, by sequence analysis, we confirmed that The SEOV strain T2016-31 existed in in Apodemus agrarius, indicating there was a phenomenon of "spillover" between virus and host in SEOV evolution.