1.Relief on fibrosis past acute rejection of the rat allograft artery from the specific 5-HT2A receptor blocker,sarpogrelate
Haihao WANG ; Ming LI ; Min WU ; Weijie ZHANG ; Zhishui CHEN ; Jun YANG
Chinese Journal of Organ Transplantation 2011;32(9):562-565
Objective To investigate the effect of sarpogrelate, a specific 5-HT2A receptor blocker,on fibrosis past acute rejection of abdominal aortic allotransplantation in rats. MethodsThe rat models of abdominal aortic transplantation were divided into three groups: allograft control group (Wistar→ SD), isograft control group ( SD→ SD) and sarpogrelate-treated group (Wistar→ SD).Sarpogrelate-treated group receivedintragastric administration of sarpogrelate every day. The pathologic and immunohistochemical findings of the transplant aorta were observed at 14th and 60th day after transplantation. ResultsAt 14th day, visible acute rejection could be observed in allograft control group,but not in isograft control group. At 60th day, vascular intimal index of sarpogrelatetreated group was significantly lower than that in allograft control group[( 16. 71 ± 3. 94)% versus (62. 41 ± 6. 54)% ,P<0. 05). The expression of PCNA and α-SMA in sarpogrelate-treated group wasalso significantly lower than that in allograft control group[(7. 37 ± 4. 61)% versus (22. 43 ±3.40)%,(8.21 ± 3. 11)% versus. (23.70 ± 2.78)%, P<0.05, respectively). Conclusion The expression of PCNA and α-SMA of the transplant aorta could be suppressed by sarpogrelate, and sarpogrelate could relieve the fibrosis past acute rejection of aortic allograft.
2.Analysis of nucleotide sequences and genetic polymorphisms of UL138 gene of low passage human cy-tomegalovirus strains in Guangzhou
Lijun CAI ; Jingjing HU ; Bo WANG ; Haihao SU ; Yuanbin WU ; Yingge MI ; Mengjie GUO
Chinese Journal of Microbiology and Immunology 2015;(6):407-413
Objective To analyze the nucleotide sequences and genetic polymorphisms of UL138 gene of low passage human cytomegalovirus ( HCMV) strains isolated from infants in Guangzhou province. Methods The low passage strains of HCMV were isolated from urine samples of 10 infants with HCMV in-fection in Guangzhou province and identified by multiplex PCR.The UL138 genes were amplified, cloned and identified with sequencing.The sequences were analyzed together with the homologous sequences of 10 clinical isolates published in GenBank.The sequences of UL138 genes were analyzed by using bioinformatics softwares for investigation of the post-translational modification sites, isoelectric points and second structures of UL138 proteins.Results Three low passage strains of HCMV ( D2, D3 and D52) were isolated from in-fants with congenital HCMV infection.The complete sequences of UL138 genes of the three strains were sub-mitted to GenBank after sequencing identification with the GenBank accession numbers of DQ180375, DQ180387 and DQ180359, respectively.The UL138 gene sequences of the three clinical isolates were high-ly conservative.Among the 841 base pairs of the UL138 gene sequences, mutations were identified in 16 sites with base substitution, no any insertion and deletion mutation was found.The 16 mutations resulted in 7 amino acid changes.No additional or deleted sites were found with regard to the post translational modifi-cation sites of UL138 protein in all clinical isolates except the Toledo strain.The isoelectric point of UL138 protein was 6.51 for all clinical isolates.Conclusion The UL138 genes and the deduced amino acid se-quences of HCMV strains isolated from infants in Guangzhou were highly conservative, regardless of the poly-morphism of UL138 gene.This study paved the way for further investigation on HCMV infection and its path-ogenic mechanism.
3.Biomechanical study of anterior transpedicular screw fixation for subaxial cervical three-column injury
Haihao WU ; Tao TANG ; Qingjiang PANG ; Xinhua YUAN ; Chunguang ZHOU
Chinese Journal of Orthopaedic Trauma 2017;19(10):897-901
Objective To compare the initial stability between anterior transpedicular screw (ATPS) fixation,anterior plate (AP) fixation and combining anterior and posterior (CAP) fixation for subaxial cervical three-column injury.Methods Six specimens of cervical spine were prepared.After measurement of the range of motion (ROM) of the intact cervical spine,the specimens were made into models of three-column injury.After the models were simulatively reconstructed using an anterior cervical cage,they were stabilized by ATPS,AP and CAP.After the ROMs of the models in the 3 fixation states were measured,the data were normalized by standardizing them to the intact state ROM which was set at 100%.The normalized ROMs of the models in the 3 fixation states were compared.Results The normalized ROMs of AP fixation state in flexion,extension,lefi lateral bending,right lateral bending,left axial rotation and right axial rotation were 119.68±8.34%,119.63±6.74%,115.20±7.91%,117.47±7.81%,120.67±5.99% and 112.35 ± 8.42%,respectively,significantly larger than those of the intact state (P < 0.05).The normalized ROMs of the other 2 states in all directions were significantly smaller than those of the intact state (P <0.05).The normalized ROM of ATPS state in flexion was 87.48 ± 5.31%,significantly larger than that of CAP state (69.60 ± 2.06%) (P < 0.05).There were no significant differences between the normalized ROMs of ATPS state and those of CAP state in extension (65.53 ± 4.36% versus 67.17 ± 3.10%),in left lateral bending (82.13 ± 2.85% versus 82.30 ±4.69%),in right lateral bending (81.78 ± 3.42% versus 81.27 ± 2.79%),in left axial rotation (83.20 ± 2.30% versus 82.95 ± 2.40%),or in right axial rotation (83.03 ± 1.30% versus 83.60 ± 6.56%) (P > 0.05).Conclusions In subaxial cervical three-column injury,the initial stability of ATPS fixation may be superior to that of AP fixation and similar to that of CAP fixation.We believe that ATPS can provide enough initial stability for subaxial cervical three-column injury.
4.Benefits and related problems of influenza immunization for health care workers
Jingjing HU ; Qiqi TAN ; Haihao SU ; Lin'gan WANG ; Yuanbin WU ; Bo WANG
Chinese Journal of Applied Clinical Pediatrics 2019;34(2):120-125
Vaccination is the most effective way to prevent influenza and severe outcoming caused by influenza viruses.Health care workers(HCW) are exposed to patients with influenza and they are at high risk of occupationally acquired influenza and of causing nosocomial infection among patients,increasing the incidence rate,the risk of severe and death of patients.Improving the influenza immunization in HCW can not only reduce the prevalence of themselves and keep a weel-oiled of health care facilities during the influenza seasons,but also reduce the risk of severe and death among patients and increase the influenza vaccine uptake in whole population.At present,the influenza immunization coverage of HCW is low.The obstacles and myths of influenza vaccine are barriers for vaccine uptake among HCW.The various strategies are critical in order to improve the influenza coverage rates of HCW.
5. Effective inhibition of human cytomegalovirus UL148 gene expression by external guide sequences in vitro
Jingjing HU ; Bo WANG ; Haihao SU ; Juncai DING ; Yuanyuan GUO ; Binhua XIE ; Yuanbin WU ; Lijun CAI ; Mengjie GUO
Chinese Journal of Experimental and Clinical Virology 2017;31(3):185-188
Objective:
To investigate the UL148 gene function of human cytomegalovirus (HCMV) low passage clinic isolate and new strategies for anti-HCMV treatment, the DNA-based external guide sequences (EGSs) were designed to inhibit UL148 RNA expression.
Methods:
UL148 RNA secondary structure was analyzed by RNA structure technique, an appropriate region was chosen for DNA-based EGS57 synthesis, targeted the UL148 RNA. The M1RNA and UL148 RNA were generated by PCR for transcription in vitro. The UL148 RNA and M1RNA were transcribed in vitro under the function of T7 RNA polymerase. The UL148 was labelled by 32P. The cleavage reactions were carried out by mixing up EGS, M1RNA with UL148 RNA for 1 h. The products were separated by urea denaturing polyacrylamide gel electrophoresis and detected with Typhoon Phosphor Imager.
Results:
UL148 RNA ranged from 58 to 72 sites was the binding position, and 57 was a cleavage site. EGS57 was designed and synthesized. EGS57 was combined with UL148 RNA to form the natural substrate of M1RNA. UL148 RNA and M1RNA were synthesized through T7 RNA polymerase catalyzing, and the products were conformed. After cleaving reactions, DNA-based EGS57 was shown to be able to cleave UL148 RNA efficiently in vitro by a complex with M1RNA.
Conclusions
UL148 RNA was cleaved efficiently by EGS57, and the cleaving site was conformed as expectation. It will provide the gene silent tool effectively for further study the function of UL148 gene.
6.In vitro expression of human cytomegalovirus UL148 RNA and prediction of its functional sites
Jingjing HU ; Yuanbin WU ; Qiqi TAN ; Haihao SU ; Juncai DING ; Yuanyuan GUO ; Binhua XIE ; Lijun CAI ; Mengjie GUO ; Bo WANG
Chinese Journal of Microbiology and Immunology 2018;38(2):94-97
Objective To express UL148 RNA of human cytomegalovirus (HCMV) clinical strains in vitro and to study its functions. Methods Urine of a newborn with HCMV infection was inocula-ted into human embryo lung cells. HCMV clinical strain was isolated and identified by multiplex PCR. UL148 gene was amplified and cloned into pGEM-T-Easy plasmid after double enzyme digestion. A recombi-nant plasmid was constructed and located at the downstream of the T7 promoter. The recombinant plasmid was identified by electrophoresis of the recombinant plasmid,PCR product and double enzyme product. Se-quencing analysis was used for final confirmation. UL148 was transcribed into RNA by 32P labeling. Post-translational modification sites were analyzed by bioinformatics method based on UL148 sequence characteris-tics. Results The clinical strain of HCMV was obtained in vitro. Electrophoresis and sequencing analysis confirmed the successful construction of the recombinant plasmid. UL148 RNA was transcribed in vitro by T7RNA polymerase. Post-translational modification sites showed that UL148 gene contained one cell adhe-sion sequence, one legume lectins beta-chain signature, two N-myristoylation sites, one casein kinase Ⅱphosphorylation site,seven protein kinase C phosphorylation sitse, one cAMP/cGMP-dependent protein ki-nase phosphorylation site, two N-glycosylation sites and one transmembrane region. Conclusion UL148 gene might encode a viral adhesion molecule involving in the signal transduction in host cells.