Objective:In this experiment,ELISA measurement was developed to detect antibody agaist PCV-2.Methods:A Cap sequence encoding for antigen epitope was obtained by PCR from the genome of porcine circovirus type 2 and then cloned into prokaryotic expression plasmids pET-22b(+).The plasmid of pET-22b(+)-cap was successfully constructed and transformed into E.coli BL21(DE3).An ELISA for detection of PCV-2 was established by high expression of Cap protein after induced by IPTG.Results:The results of Western blot showed that the expressed protein was 30 kD and possessed immunological activity.Positive rate was 73.1% and negative rate was 26.9% respectively for 643 serum samples which were examined with the measurement by ELISA.Conclusion:The ELISA method is applicable in clinic.

OBJECTIVETo evaluate the effect of integrative medical treatment (IMT) with serial Chinese recipes on quality of life (QOF) of rehabilitation stage in SARS patients.

METHODSEighty-five SARS patients of rehabilitation stage were enrolled in the clinical study. They were divided into the IMT group (62 patients received serial Chinese recipes and western medicine) and the control group (23 patients received western medicine alone). The serial Chinese recipes were given according to patients' syndrome, one dose per day for oral intake for 3 weeks. QOF scoring in patients was observed.

RESULTSQOF scoring in the IMT group before treatment was not significantly different from that in the control group. After 3 weeks treatment, it improved to some extent in both groups, but the improvement in the IMT group was superior to that in the control group in respect of total score and score of psychologic emotional factors.

CONCLUSIONSerial Chinese recipe could improve QOF of rehabilitation stage in SARS patients.

Adolescent ; Adult ; Aged ; Convalescence ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Ofloxacin ; therapeutic use ; Phytotherapy ; Quality of Life ; Ribavirin ; therapeutic use ; Severe Acute Respiratory Syndrome ; drug therapy ; immunology ; rehabilitation

Objective To evaluate the effect of local mild hypothermia on the expression of phosphatase and tensin homolog deleted on chromo-some ten (PTEN) protein during global cerebral ischemia-reperfusion (I/R) in rats.Methods One hundred and eight healthy adult male Wistar rats,weighing 230-280 g,were randomly divided into 3 groups (n =36 each) using a random number table:sham operation group (group S),cerebral I/R group (group I/R) and mild hypothermia cerebral I/R group (group HI/R).Global cerebral I/R was induced by 4-vessel occlusion method described by Pulsinelli.Bilateral vertebral arteries were permanently occluded by cauterization,and bilateral carotid arteries were occluded for 15 min.Nasopharyngeal cooling was applied and the nasopharyngeal temperature was reduced to 32.5-33.5 ℃ and maintained at this level for 1 h.When the nasopharyngeal temperature was reduced to 32.5-33.5 ℃,the bilateral carotid arteries were clamped in group HI/ R.Six rats were chosen to be anesthetized and sacrificed immediately before ischemia and at 4,8,12,24 and 72 h of reperfusion.Hippocampal specimens were obtained to detect the expression of phosphorylated PTEN (pPTEN),Bcl-2 and Bax protein (by immunohistochemistry) in hippocampal CA1 region,contents of S100B protein (by ELISA) and MDA (by thiobarbituric acid method),and activity of SOD (by xanthine oxidase method).Results Compared with group S,the expression of p-PTEN and Bcl-2 protein,ratio of Bcl-2/Bax protein and activity of SOD were significantly decreased,and the expression of Bax protein and contents of MDA and S100B protein were increased after reperfusion in group I/R (P ＜ 0.05),and the ratio of Bcl-2/Bax protein was decreased,and there was no significant difference in the expression of p-PTEN,Bcl-2 and Bax protein,activity of SOD and contents of MDA and S100B protein after reperfusion in group HI/R (P ＞ 0.05).Compared with group I/R,the expression of p-PTEN and Bcl-2 protein,Bcl-2/Bax ratio and activity of SOD were significantly increased and the expression of Bax protein and content of MDA and S100B protein were decreased after reperfusion in group HI/R (P ＜ 0.05).Conclusion The mechanism by which local mild hypothermia reduces cerebral damage is related to inhibition of activation of PTEN protein in the brain tissues during global cerebral I/R in rats.

Objective To determine the serum concentration of MCP-1 and the expression of MCP-1 protein in the pancreas in the piglet with chronic obstructive pancreatitis and to explore the role of MCP-1 protein in pancreatic fibrosisits.Methods The piglet model of chronic obstructive pancreatitis was established by incomplete ligation of the pancreatic duct.The piglets were sacrificed at 4, 6, 8 weeks after induction.Pathological changes of pancreas were examined.Pancreatic fibrosis was assessed by VG staining.Serum MCP-1 concentrations were detected by ELISA method.MCP-1 and α-SMA, PDGF, TGF-β1 and NF-κB protein expression were detected by immunohistochemistry.Results The induction was successful in 14 piglets ( 58.3% ).Mild atrophic changes, interstitial fibrosis, chronic inflammatory cell infiltration could be observed in the body and tail of pancreas from the 4th week in the experimental group.The most obvious changes occurred in the 8th week.Stage Ⅰ pancreatic fibrosis occurred in 5 piglets (35.7%), stage Ⅱ in 4 piglets (28.6%), stage Ⅲ in 5 rats ( 35.7% ).Seurm MCP-1 at 4, 6, 8 weeks was ( 102.44 ± 36.25 ) pg/ml,(97.84 ± 28.67) pg/ml, ( 94.32 ± 28.42 ) pg/ml, respectively, and was significantly higher than that in control group [ ( 10.42 ±5.86) pg/ml, (8.58 ±4.86) pg/ml, (8.22 ±4.58) pg/ml, P ＜0.01 ].There was no MCP-1 protein expression in the control group;MCP-1 protein was detected in the successful induction group, and MCP-1 expression was positively correlated with expressions of the PDGF, TGF-β1, α-SMA and NF-κB.Conclusions MCP-1 may play an important role in the course of pancreatic fibrosis in chronic obstructive pancreatitis.

Objective: To analyze the epidemiological characteristics of coronavirus disease 2019 (COVID-19) local infections in Zhejiang Province during the period between March and May, 2022, so as to provide the evidence for COVID-19 control. Methods: The individual investigation data and field epidemiological investigation data of local COVID-19 cases in Zhejiang Province from March to May 2022 were extracted from the National Diseases Prevention and Control Information System of China. The temporal, spatial and population distribution, identification and booster vaccination of local COVID-19 cases were analyzed using a descriptive epidemiological method. Factors affecting the severity of clinical symptoms were identified among local COVID-19 cases using a multivariable logistic regression model. Results: A total of 289 local COVID-19 epidemics occurred in Zhejiang Province from March to May 2022, and all infections were caused by the Omicron variant. A total of 1 598 local COVID-19 infections were reported, including 672 confirmed cases (42.05%) and 926 asymptomatic cases (57.95%), and the 672 confirmed case included 614 mild cases (91.37%), 58 normal cases (8.63%), while no severe, critically ill or dead cases were reported. The mean duration of COVID-zero community, mean duration of COVID-19 epidemics and mean length to COVID-19 peak calculated according to 7 epidemics with 50 and more cases were (6.14±1.07), (13.43±4.39), (3.00±1.63) d, respectively. COVID-19 infection was predominantly detected in centralized quarantine sites (1 105 cases, 69.15%), and 843 cases completed booster immunization of COVID-19 vaccines (52.75%). Multivariable logistic regression analysis showed that age (with ages of 60 years and older as the reference, age of 6 to 17 years, OR=0.111, 95%CI: 0.024-0.508; age of 18 to 44 years, OR=0.341, 95%CI: 0.163-0.713) and booster COVID-19 vaccination (OR=0.219, 95%CI: 0.117-0.410) were protective factors for developing more severe clinical symptoms following COVID-19 infections. Conclusions All local COVID-19 infections were caused by Omicron variant in Zhejiang Province during the period between March and May 2022, and all cases had mild clinical symptoms. Preschool children, middle-aged and elderly residents and individuals that did not complete booster COVID-19 vaccination may had relatively more severe clinical symptoms following COVID-19 infections.

Objective To investigate the predictive value of androge receptors in stage pT1 non-muscle-invasive bladder cancer.Methods A total of 196 patients with stage pT1 non-muscle-invasive bladder cancer who underwent a transurethral resection of the bladder in Baoji People's Hospital from February 2003 to June 2012 were recruited to carry a retrospective analysis.The mRNA expression of the androge receptors transcript variants 1 (AR1) and 2 (AR2) was measured by quantitative real-time PCR.Spearman's correlation coefficient was used to analysis the correlation of androge receptors mRNA level to KRT5 and KRT20 mRNA.Results Kaplan-Meier analysis indicated that the recurrence-free survival,progression-free survival and cancer specific survival of patients with high AR1 mRNA expression (≥35.47) were significant better than patients with low AR1 mRNA expression (P ＜0.05).Multivariate COX regression analysis revealed that high AR1 mRNA expression was an independent prognostic marker for recurrence-free survival and cancer specific survival (P ＜ 0.05).Spearman rank correlation revealed a significant positive association between mRNA expression of AR1 and KRT5 (rs =0.3171,P ＜ 0.001) as well as a negative association with multifocal tumors (rs =0.1478,P ＜ 0.05).Conclusion Androge receptors mRNA expression can predict recurrence-free survival and cancer specific survival in patients with stage T1 nonmuscle-invasive bladder cancer.Further studies on the levels of androge receptors mRNA tend to be particularly important.

BACKGROUND: The restenosis occurs up to 20%-30% following metal coronary stent implantation. Under the support of the 863 program, the feasibility to treat coronary artery stenosis using a novel drug-eluting stent (DES) has been investigated to reduce restenosis. OBJECTIVE: A drug-eluting stent (rapamycin as drug mode) was implanted into porcine models of coronary stenosis. The safety and efficacy of the drug-eluting stent were observed and compared with bare-metal stent. DESIGN, TIME AND SETTING: A randomized controlled animal experiment was performed in the Fu Wai Hospital for Cardiovascular Disease between November 2003 and April 2004. MATERIALS: A novel bioinspired phospholipid copolymer was synthesized by free radical polymerization of stearyl methacrylate, β-hydroxypropyl methacrylateand 3-(trimethoxysilyl) propylmethacrylate. METHODS: Twenty-one pigs were randomly divided into 3 groups: bare-mental stent, drug-eluting stent, and polymer-coated stent. The treated stents pre-loaded onto a delivery system through the use of crimping instrument were implanted into pig's coronary artery, with 2 stents per pig. MAIN OUTCOME MEASURES: Determination of luminal diameter, luminal area, mean intimal thickness on and between the stents, neointimal area, percentage of luminal area restenosis, and damage index using an image analysis instrument. RESULTS: At 28 days after implantation, there was significant difference in mean intimal thickness on and between the stents, as well as neointimal area, between the DES and bare-metal stent groups (P < 0.05). The neointimal area was reduced by 44.87% in the DES group compared with the bare-metal stent group. No significant difference in percentage of luminal area restenosis was found between the DES and bare-metal stent groups, but P value equaled to 0.053, which was close to 0.05. In addition, no restenosis was found in the DES group. CONCLUSION: Rapamycin DES can markedly resist intravascular intimal hyperplasia and restenosis following stenting.

Objective To investigate the related risk factors of cerebral microbleeds (CMBs) and their relations with leukoaraiosis and lacunar infarction in patients with cerebrovascular disease.Methods One hundred and sixty-four patients with cerebrovascular disease,admitted to our hospital from January 2015 and January 2017 were chosen;susceptibility weighted imaging (SWI) was performed,and the distribution and number of CMBs were recorded by Microbleed Anatomical Rating Scale (MARS).The demographic data,personal history,previous medical history,biochemical indices and imageological examination results of the patients were collected and multivariate Logistic regression analysis was carried out on the basis of these data.Spearman dependence analysis was used to analyze the relations of CMBs with leukoaraiosis and lacunar infarction.Results Age (OR=1.071,P=0.024,95％ CI:1.009-1.137),hypertension (OR=3.875,P=0.012,95％CI:1.347-11.148),leukoaraiosis(OR=2.080,P=0.005,95％ CI:1.245-3.475),lacunar infarction (OR=2.326,P=0.003,95％ CI:1.336-4.050) were independently associated with CMBs.And number of CMBs had positive correlation with severity of leukoaraiosis and number of lacunar cerebral infarcts (r=0.564,P=0.000;r=0.762,P=0.000).Conclusion Age,hypertension,leukoaraiosis,and lacunar infarction are the independent risk factors of CMBs;the severity of CMBs is positively correlated with severity of leukoaraiosis and lacunar infarction.

This study was purposed to construct a fusion DNA vaccine containing WT1 multi-epitope and stimulating epitope of mycobacterium tuberculosis heat shock protein 70 and to detect its expression and immunogenicity. On the basis of published data, a multi-epitope gene (Multi-WT1) containing three HLA *0201-restricted CTL epitopes: one HLA*2402-restricted CTL epitope, two Th epitopes and one universal Th Pan-DR epitope (PADRE) was constructed. DNA-coding sequence was modified by Computer-Aided Design (CAD) to optimize proteasome-mediated epitope processing through the introduction of different amino acid spacer sequences. The synthetic nucleotide sequence was then inserted into an eukaryotic vector to construct the plasmid pcDNA3.1-WT1.For enhancing CTL activity, HSP70 fragment including stimulatory domain P407-426 was amplified by PCR from mycobacterial HSP70 gene and cloned into pcDNA3.1(+). Then Multi-WT1 was fused to the N-terminal of pcDNA3.1-mHSP70(407-426) to make the multi-epitope fusion gene vaccine pcDNA3.1-WT1-mHSP70(407-426). HEK-293T cells were transfected with this vaccine and the expressed product was identified by RT-PCR. Enzyme-linked immunospot assay (ELISPOT) was used to evaluate the immunological responses elicited by vaccine. The results showed that the most of WT1 epitopes could be correctly cleaved which was confirmed by software Net Chop 3.1 and PAPROCIanalysis. RT-PCR showed correct expression of target gene in HEK293T cells and ELISPOT showed specific T-cell responses. It is concluded that the eukaryotic expression vector PcDNA3.1-WT1-mHSP70(407-426) fusion gene has been successfully constructed and the immunity response is also elicited, which is a good candidate for further research of DNA vaccine.
Bacterial Proteins ; genetics ; immunology ; Epitopes ; genetics ; immunology ; Genetic Vectors ; HSP70 Heat-Shock Proteins ; genetics ; immunology ; Humans ; Immunodominant Epitopes ; Vaccines, DNA ; genetics ; immunology ; WT1 Proteins ; genetics ; immunology

Objective To analyze the efficacy of different doses of intravenous immunoglobulin (IVIG) in the treatment of acquired severe aplastic anemia (AA) in children. Methods The clinical data of hospitalized children with severe AA who received adjuvant immunosuppressive therapy of IVIG from January 2000 to December 2015 were retrospectively analyzed. According to different doses of treatment, the children were divided into low dose group ( IVIG 200-400 mg/ (kg·d) once every 4 weeks for 6 times), high dose group (IVIG 1 g/ (kg·d ) x 2 days once every 4 weeks for 6 times). Results All the children were followed up until December 31, 2015. Among the 61 children, it was effective in 41 children and total effective rate was 67.2%. The effective rate of anti thymocyte globulin (ATG) treatment in high dose group was higher after 3 months than that of low dose group, and there was statistical difference (P=0.020). The interval between first dose of IVIG and first dose of ATG in 20 cases of ineffectiveness was 2.0 (2.0-5.0) d, while that in 41 cases of effectiveness was 8.0 (7.0-9.0) d, and the difference is statistically significant (P<0.001); Among the 20 ineffective children, 18 children had the interval <7 day. The survival rates of the two groups were 80% and 87.1%, respectively, and there was no difference between two groups (P>0.05). The incidence of severe infections in the high-dose group was lower than that in the low-dose group after the use of ATG for 6 months, and there was statistical difference (P=0.008). Conclusions High dose of IVIG therapy can increase the early response rate in children with acquired severe AA, but it does not increase the long-term effectiveness, cure rate and 5 year survival rate. In addition, it can reduce the severe infection rate, but cannot reduce the total infection rate and infection related mortality rate.