1.Isolation and Complete Genomic Sequence Analysis of a New Sindbis-like Virus
Jingjing WANG ; Hailin ZHANG ; Yanchun CHE ; Lichun WANG ; Shanghui MA ; Longding LIU ; Yun LIAO ; Qihan LI
Virologica Sinica 2008;23(1):31-36
The Sindbis-like virus was first discovered in China in 1986. Its complete genomic sequence consists of more than 11 000 bp encoding more than 3 700 amino acids. It contains a 5' non-transcriptional region (5'-NTR) in a non-structural region, four non-structural proteins (nsP1, nsP2, nsP3, nsP4) regions, capsids in conserved and non-conserved regions and structural E1, E2, E3, 6K regions and a 3' non-transcriptional region (3'-NTR). The Sindbis-IMB was isolated from the blood of a patient suspected to have encephalitis, and was followed by identification and passage. The virus RNA was extracted from virus supernatant in infected cells and the whole genome was divided into 12 fragments; RT-PCR was then performed to amplify the 12 fragments for complete sequencing. The results showed that the whole genomic sequence of Sindbis-IMB consists of 11 717 bp encoding 3 773 amino acids. Homology comparison with other Sindbis-like isolates demonstrated the highest similarity was the YN87448 with a variation of 1% strain isolated in Yunnan Province and the second highest to the SAAR86 strain with a variation of~1.2%.The nucleotide sequence variations were present in non-structural regions, resulting in amino acids K, E, N, R, H, and L in protein sequences in positions 230, 231, 443,781, 1 582, and 1746 in the new isolation respectively. Furthermore, three additional amino acids--glutamic acid, serine and alanine--were noted in nsp4 terminus as compared to the YN87448 isolate.
2.Induced pluripotent stem cells can be reprogrammed from human umbilical cord mesenchymal cells by six transcription factors
Hanhua YANG ; Yuanguo CHEN ; Xiulan LAI ; Lichun XIE ; Zexin ZHENG ; Xuewu JIANG ; Lian MA
Chinese Journal of Applied Clinical Pediatrics 2014;29(17):1331-1336
Objective To establish and identify the induced pluripotent stem cell(iPSC) line reprogrammed from human umbilical cord mesenchymal cells(HuMSCs).Methods HuMSCs were cultured by adhesion method,and OCT4,SOX2,KLF4,c-Myc,NANOG,LIN-28 were transfected into HuMSCs with lentiviral victor to reprogramme HuMSCs into iPSC.Morphological observation,pluripotency genes (SOX2,TDGF1,THY-1,OCT4,REX1 and TERF1) expression,alkaline phosphatase detection,karyotype analysis,embryonic stem cells (ESC) specific proteins (NANOG,OCT4,SSEA-4,TRA-1-81) immunofluorescence staining,differentiated into teratomas in vivo(inject the iPSC into SCID mice) and embryniod bodies in vitro were performed to exam the pluripotency of the iPSC.Results Four days after being infected by lentivirus,the HuMSCs became round-shape; 10 days after infection,some embryonic stem(ESC)-like colonies appeared.Fourteen days after infection,picked up the regularly shaped colonies and cultured several passages.About 1.25% HuMSCs were reprogrammed into iPSC.The iPSC presented clone-like growth like ESC.All the cells were positive to alkaline phosphatase staining and expressed the pluripotency genes.The iPSC also expressed the ESC specific proteins,and karyotype analysis showed normal chromosome caryotype (46,XY).Furthermore,the iPSC could form embryoid bodies in vitro,expressed alpha fetoprotein(AFP),smooth muscle actin(SMA) and β-tubulin.The iPSC could alsoform teratomas in vivo.Conclusion OCT4,SOX2,KLF4,c-Myc,NANOG,LIN-28 can reprogram HuMSCs into iPSC efficiently.
3.Effect of anti-CD59 on CVF-induced platelet activation
Xinxue LIAO ; Lichun WANG ; Hong MA ; Shoujian HUANG ; Weiyi MAI ; Chon FENG
Chinese Journal of Pathophysiology 1989;0(05):-
AIM: To study the reactions of human platelet to active complement and the effects of anti-CD59 on human platelet activation induced by complement. METHODS: By applying CVF to activate complement, the platelet aggregation and release reactions induced by activated complement with or without appling anti-CD59 with different doses to block the complement modulative protein CD59 in healthy individuals, were observed. RESULTS: CVF induced platelet release and significant and lasting metamorphosis in healthy individuals, but platelet aggregation was not observed. CVF-induced platelet metamorphosis showed positive linear correlation to lg concentration of CVF (r=0 970. P
4.Correlation between the effect of angiotensin-(1-7) on cardiac hypertrophy and extracellular signal-regulated kinase in pressure-overloaded rats
Jiangui HE ; Yiyi HUANG ; Hong MA ; Xiaohong HE ; Xinxue LIAO ; Lichun WANG ; Jun LIU
Chinese Journal of Pathophysiology 1999;0(09):-
AIM: To investigate the effects and mechanisms of angiotensin-(1-7) on cardiac hypertrophy in pressure-overloaded rats. METHODS: Ar at model of pressure-overloaded heart was induced by constriction of abdominal aorta. Seventy-five male Sprague Dawley rats were randomized to sham-operated group, model control group and angiotensin-(1-7) treatment group. They were treated with intravenous infusion of angiotensin-(1-7) (25 microgram/kg per hour) or saline by minipump. RESULTS: Abdominal aortic banding resulted in a significant increases in LVW/BW, myocardial angiotensinⅡlevels, and p-ERK1/2 expression. Angiotensin-(1-7) had no effect on aortic banding-induced increases in myocardial angiotensinⅡlevels, but it significantly attenuated aortic banding-induced increases in LVW/BW and p-ERK1/2 expression. CONCLUSION: Angiotensin-(1-7) attenuates the development of cardiac hypertrophy in pressure-overloaded rats. It may be associated with the inhibition of p-ERK1/2 expression in cardiac tissue.
5.Isolation and Complete Nucleotide Sequence of the Measles Virus IMB-1 Strain in China
Shaohui MA ; Lichun WANG ; Jiansheng LIU ; Haijing SHI ; Longding LIU ; Qihan LI
Virologica Sinica 2010;25(6):381-389
The complete nucleotide sequence of the measles virus strain IMB-1, which was isolated in China, was determined. As in other measles viruses, its genome is 15,894 nucleotides in length and encodes six proteins. The full-length nucleotide sequence of the IMB-1 isolate differed from vaccine strains (including wild-type Edmonston strain) by 4% -5% at the nucleotide sequence level. This isolate has amino acid variations over the full genome, including in the hemagglutinin and fusion genes. This report is the first to describe the full-length genome of a genotype H 1 strain and provide an overview of the diversity of genetic characteristics of a circulating measles virus.
6.The clinical significance of soluble thrombomodulin in acute coronary syndrome
Xinxue LIAO ; Xin LI ; Lichun WANG ; Jiangui HE ; Xugang DONG ; Zhimin DU ; Hong MA
Chinese Journal of Postgraduates of Medicine 2008;31(21):7-9
Objective To study the levels of soluble thrombomodulin (sTM)in patients with acute coronary syndrome (ACS) and evaluate its clinical significance. Method Measured the sTM levels with enzyme linked immunosorbent assay, and described the characteristics of coronary arteriography, risk factors of coronary heart disease, and adverse events in a case-control study of 48 ACS patients (ACS group)and 10 normal people (control group). Results The level of sTM in ACS group was (3.67±1.71) μg/L, and (2.34±0.43)μg/L in control group (P<0.05). The level of sTM in the patients of risk factors or impaired vessels number more than 2 increased significantly than those in the patients of risk factors or impaired vessols number inferior or equal to 2, (4.93±2.76) μg/Lvs (3.13±0.81) μg/L, P<0.05, (4.60± 2.83) μg/L vs (2.91±0.23) μg/L, P < 0.05 respectively. The incidence of cardiac events in the patients of sTM more than 3.2 μg/L (70.0%)was higher significantly than that in the patients of sTM inferior or equal to 3.2 μg/L(35.7%), P< 0.05. Conclusions The levels of sTM are valuable markers to evaluate the impaired degree and scope of endothelial cells in ACS. They are also associated with the number of risk factors, and useful in predicting the extent and prognosis of the disease.
7.Complete Nucleotide Sequence of a Mumps Virus SP Strain Isolated in China
Shaohui MA ; Jiansheng LIU ; Haijing SHI ; Lichun WANG ; Jingjing WANG ; Longding LIU ; Qihan LI
Virologica Sinica 2009;24(1):28-36
The complete nucleotide sequence of the mumps virus SP, which was isolated in China, was determined. As with other mumps viruses, its genome was 15 384 nucleotides (nts) in length and encoded seven proteins. The full-length nucleotide sequence of the SP isolate differed from other strains by 4%-6.8% at the nucleotide sequence level. Due to variations of amino acids over the full genome (including the HN and N genes), this isolate exhibited significant variations in the antigenic sites. This report is the first to describe the full-length genome of a genotype F strain and provide an overview of the diversity of genetic characteristics of a circulating mumps virus.
8.Effect of Parasep? feces centrifuge tube method on detecting schistosome eggs
Nian MA ; Huaming ZHANG ; Xiong LIU ; Chuanyun XIAO ; Xiaohong WEN ; Xia LI ; Lichun DONG ; Caixia CUI ; Zuwu TU
Chinese Journal of Schistosomiasis Control 2014;(4):431-433
Objective To evaluate the effect of the Parasep? feces centrifuge tube method on detecting schistosome eggs. Methods A total of 803 residents aged from 6-65 years were selected in 2 schistosomiasis endemic villages Jiangling Coun-ty Hubei Province and their stool samples were collected and detected parallelly by the Kato-Katz technique nylon silk egg hatching method and Parasep? feces centrifuge tube method at the same time. Results Among the 803 people 15 cases were found of schistosome egg positive and the positive rate was 1.87%. The positive rates of the Kato-Katz technique nylon silk egg hatching method and Parasep? feces centrifuge tube method were 0.75% 1.49% and 1.12% respectively. The schistosome eggs got with the Parasep? feces centrifuge tube method were clear and easy to identify. Conclusion In low endemic areas of schistosomiasis the Parasep? feces centrifuge tube method can be used as schistosomiasis japonica etiology diagnosis method.
9.A Comparison of the Biological Characteristics of EV71 C4 Subtypes from Different Epidemic Strains
Lichun WANG ; Songqing TANG ; Yanmei LI ; Honglin ZHAO ; Chenghong DONG ; Pingfang CUI ; Shaohui MA ; Yun LIAO ; Longding LIU ; Qihan LI
Virologica Sinica 2010;25(2):98-106
The comparative analysis of the biological characterization and the genetic background study of EV71 circulating strains is commonly recognized as basic work necessary for development of an effective EV71 vaccine. In this study, we sequenced five EV71 circulating strains, isolated from Fuyang, Hefei, Kunming and Shenzhen city of China and named them FY-23, FY-22, H44, K9 and S1 respectively. The sequence alignment demonstrated their genotypes be C4. The genetic distance of the VP1 gene from these isolates suggested that they were highly co-related with genetic identity similar to other previously reported EV71 strains in China. Additionally, these strains were identified to display some obvious proliferation dynamics and plaque morphology when propagated in Vero cells. However, a distinctive difference in pathogenic ability in neonatal mice was found. Some differences in cross neutralization test & immunogenic analysis were also found. All these results are related to the biological characterization of circulating EV71 strains in China and aid in the development of an EV71 vaccine in the future.
10.Physical Activity and Vascular Dilation Function in Healthy Middle-aged Individuals
Qi LIANG ; Donghong LIU ; Yuling WANG ; Bing SUN ; Fengqiao LIN ; Hanjing GAN ; Guifu WU ; Lichun WANG ; Hong MA
Chinese Journal of Rehabilitation Medicine 2009;24(6):489-493
Objective: Vascular dilation dysfunction has been linked with risk of cardiovascular disease. This study was undertaken to investigate the relationship between physical activity and vascular dilation function in healthy middle-aged adults to help explaining the effect of physical activity on preventing cardiovascular disease. Method: We recruited 91 healthy middle-aged adults to complete a serf-report 7-day physical activity recall questionnaire and an exam of brachial artery flow-mediated dilation(FMD) and Nitroglycerin-mediated dilation (NMD) detected by ultrasound. The relationship between physical activity level (PAL) and FMD and NMD were explored. Result: Physical activity showed a significant and positive relationship with the brachial artery FMD, even after adjustment for possible confounding factors (r=0.363, P<0.01). The group of high physical activity level had the highest FMD. The means of FMD (%) in low, moderate and high PAL groups were respectively 8.70%±3.93%,9.01%±3.22% and 12.38±5.67% with significant difference between individuals of low and high PAL and between moderate and high PAL group(P<0.01). The means of NMD (%) is 25.13%±6.52%,24.38%±5.44% and 29.50%±7.25% respectively (P> 0.05) and there was no significant difference among three groups. There was no positive relation between PAL and FMD in premenopausal females but in men and postmenopausal females. Although individuals of high PAL have the best FMD, the moderate PAL can also retard FMD decrease with ageing. Conclusion: Maintaining high physical activity level can enhance endothelial-dependent vascular dilation, and moderate or high physical activity level can prevent endothelial-dependent vaseular dilation declining with aging, which may contribute to decrease risk of cardiovascular disease in healthy middle-aged adults.