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.Prevalence and risk factors of restless leg syndrome in hemodialysis patients after kidney transplantation failure
Houzhen TUO ; Xiaoyang MA ; Jingjing CHE ; Zelong TIAN ; Yinong CUI ; Kui CHEN ; Yun XUE ; Yongbo ZHANG
Chinese Journal of Neuromedicine 2017;16(8):844-848
Objective To investigate the prevalence and risk factors of restless leg syndrome (RLS) in hemodialysis patients after kidney transplantation failure.Methods Patients of hemodialysis after kidney transplantation failure were investigated by face-to-face interviews,from March to July,2015,at four dialysis units in Beijing.RLS was diagnosed according to the International RLS Study Group (IRLSSG) criteria.The severity of RLS was assessed using International RLS Rating scale.Besides,three validated sleep disorder questionnaires (Hamilton anxiety and depression scale,Epworth sleepiness scale and Pittsburgh sleep quality index) were completed by the patients at the same time.Results Ninety-four hemodialysis patients after kidney transplantation failure were enrolled;46 patients (48.94%) met the diagnosis of RLS,the average age was 53.44±l 1.89 years,and the median time of RLS onset after kidney transplantation failure was 46 months.The International RLS Rating scale scores of the patients were 17.26±7.81;76.0% patients were above moderate.As compared with the non-RLS patients,patients with RLS used more erythropoietin (44/48 vs.46/46),less ferrila (30/48 vs.19/46),and few hypnotic medicine (10/48 vs.3/46),with significant differences (P<0.05).The serum ion,serum ferritin and serum Vitamin B12 of patients with RLS were significantly lower as compared with non-RLS patients (P<0.05);and poorer sleep quality and higher depression scale scores in the patients with RLS were noted as compared with those in the non-RLS patients (P<0.05).Conclusion The prevalence of RLS in hemodialysis patients after kidney transplantation failure is high,low iron protein content,low serum iron content and low vitamin B12 levels may be risk factors for RLS.
3. The expression of PGC-1α in carotid atherosclerotic plaque induced by cytomegalovirus
Huiqi WANG ; Jingjing CHE ; Houzhen TUO ; Zijing FENG ; Jun CHEN
Chinese Journal of Experimental and Clinical Virology 2019;33(2):171-174
Objective:
To investigate the role of peroxisome proliferator-activated-receptor-γ coactivator-1α (PGC-1α) in carotid atherosclerosis due to human cytomegalovirus (HCMV) infection.
Methods:
Thirty-three samples of carotid arterial sclerosis plaques after carotid endarterectomy (CEA) were collected in the experiment group, and 12 pieces of normal intracranial arteries were collected in the control group. The plaques of carotid artery were studied using in situ hybridization (ISH) and immunohistochemistry (IHC) for the role of PGC-1α in HCMV related atherosclerosis. HCMV immediate early (IE) gene of two groups was detected using ISH, and the expression of PGC-1α using IHC.
Results:
HCMV IE gene was positive 72.7% in the atherosclerotic plague group, while 16.6% in the control group, there was significant difference between the two groups (
4. Role of HCMV infection in the activation and oxidative stress of cultured human umbilical vein endothelial cells
Houzhen TUO ; Jingjing CHE ; Lei LIU ; Jiawei WANG ; Zijing FENG ; Jun CHEN ; Jimei LI ; Yongbo ZHANG ; Dexin WANG
Chinese Journal of Experimental and Clinical Virology 2017;31(4):298-301
Objective:
To investigate the role of the activation and oxidative stress of cultured human umbilical vein endothelial cells (HUVEC) after HCMV infection.
Methods:
HUVECs were divided into four groups: control, HCMV(+ ), after HCMV AD169 infection, and the supernatant of the culture was extracted, and the cells were lysed. The levels of vascular cellular adhesion molecule-1 (VCAM-1) in HUVEC were measured by real-time PCR. And the content of nitrogen monoxide (NO) of the supernatant was detected by nitrate reductasemethod accordingly.
Results:
Twenty-four hours after infection, the mRNA expression of VCAM-1 in HUVECs of HCMV infected group increased obviously compared to control, and NO quantity increased accordingly and time-dependently. There was significant difference between groups(
5. The role of lectin-like Ox-LDL receptor-1 on endothelial cell injury caused by human cytomegalovirus infection
Jingjing CHE ; Huiqi WANG ; Yitong DU ; Xiaojiao XU ; Li YI ; Ruijin WANG ; Jun CHEN ; Lei LIU ; Dexin WANG ; Houzhen TUO
Chinese Journal of Experimental and Clinical Virology 2019;33(5):500-503
Objective:
To investigate the role of lectin-like Ox-LDL receptor-1( LOX-1) in the activation and oxidative stress of cultured human umbilical vein endothelial cell(HUVEC) after human cytomegalovirus(HCMV) infection.
Methods:
HUVEC were divided into four groups: HCMV, Control, Carrageenan, and HCMV+ Carrageenan. After HCMV AD169 infection, the supernatant of the culture was extracted, and cells were lysed. The levels of LOX-1 mRNA, intercellular adhesion molecule-1(ICAM-1) mRNA and vascular cellular adhesion molecule-1(VCAM-1) in HUVEC were measured by real-time PCR. And the content of nitrogen monoxidum(NO) of the supernatant was detected by nitrate reductase method accordingly.
Results:
24 h after infection, the mRNA expression of LOX-1, ICAM-1 and VCAM-1 in HUVEC of HCMV infected group increased obviously compared to control, and NO quantity increased accordingly. The mRNA expression of LOX-1, ICAM-1 and VCAM-1 and the quantity of NO decreased after adding the LOX-1 inhibitor carrageenan. There was significant difference between groups(