Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 55
Filter
Add filters








Year range
1.
Article in Chinese | WPRIM | ID: wpr-920829

ABSTRACT

@#Surgery is a classic traditional method for the treatment of early-stage esophageal cancer, and it is also recognized as an effective first-choice method in the medical community. With the development of endoscopic technology, esophagus-preserving comprehensive treatment of esophageal cancer has almost the same or even better effects in some aspects in the treatment of early esophageal cancer than surgery. Many clinical guidelines have also recommended it as the first-choice treatment for early esophageal cancer. The room for surgical treatment of esophageal cancer has been further compressed. This article discusses the comprehensive treatment model of esophageal cancer from the perspective of thoracic surgery, aiming to find a new position of thoracic surgery in the treatment of esophageal cancer.

2.
Chinese Journal of Lung Cancer ; (12): 613-622, 2021.
Article in Chinese | WPRIM | ID: wpr-888588

ABSTRACT

BACKGROUND@#Lung cancer has the highest mortality in China. Different treatments are of great significance to the prognosis of patients. By comparing stage Ia non-small cell lung cancer (NSCLC) patients' survival rates for ablation and for sub-lobectomy, we studied the difference in the effects of the two treatments on patient prognosis.@*METHODS@#Using the Surveillance, Epidemiology, and End Results (SEER) database, we screened eligible patients with stage Ia NSCLC from January 2004 to December 2015. Then, 228 patients treated with ablation and 228 patients treated with sub-lobotomy were then selected based on propensity score matching. After stratification, matching, and adjustment the Kaplan-Meier analysis was performed to compare the overall survival rates of patients treated with the two procedures.@*RESULTS@#The Kaplan-Meier survival analysis showed that there is a significant difference between the ablation group and the sub-lobectomy group (P<0.05). In the univarlable analysis, the hazard ratio (HR) of sub-lobotomy group was 0.571 (95%CI: 0.455-0.717) compared with the ablation group. Patients treated with sub-lobectomy had a 0.571 times greater risk of adverse outcomes than those treated with ablation. In the multivariable analysis, the HR for sub-lobectomy group was 0.605 (95%CI: 0.477-0.766) compared with the ablation group. Patients treated with sub-lobectomy had a 0.605 time greater risk of adverse outcomes than those treated with ablation. The results suggested that the overall survival rate of patients with stage Ia NSCLC treated with sub-lobotomy was higher than that of patients treated with ablation.@*CONCLUSIONS@#This study suggests that there is a significant difference in overall survival of stage Ia NSCLC patients treated with ablation and with sub-lobotomy. Patients treated with sub-lobotomy for stage Ia NSCLC had higher overall survival than those treated with ablation.

3.
Article in Chinese | WPRIM | ID: wpr-804622

ABSTRACT

Objective@#To sequence the 3′UTR of enterovirus 71 strains, investigate its foundation and impact in virulence by constructing a 3′UTR-replaced recombinant cDNA infectious clone.@*Methods@#Viral RNA of EV-A71 isolated viruses were extracted, and the nucleotide analysis was performed after sequencing. The 3′UTR of a full-length infectious clone of SDLY107 strain was replaced by its corresponding part of SDLY1 strain, and then the recombinant virus was constructed and identified.@*Results@#The nine isolated strains were classified into sub-genotype C4a of enterovirus (EV)-A71 by analysis, and nucleotide sequence homology for 3′UTR were 94%-100%. 3′UTR of EV-A71 strains may be associated with its pathogenicity. Identification of the rescued virus by sequencing and indirect immunofluorescence confirmed the successful construction of infectious recombinant virus.@*Conclusions@#Sequence analysis indicated that the 3′UTR may be involved in the pathogenicity of EV-A71. The recombinant virus SDLY107(1-3′UTR) was rescued successfully. Our study may provide evidence for further research on the influence of 3′UTR on the virulence of enterovirus 71.

4.
Article in Chinese | WPRIM | ID: wpr-805141

ABSTRACT

Objective@#To investigate the relationship between enterovirus 71 (EV71) and autophagy and the effect of autophagy on viral replication.@*Methods@#RD cells were pretreated with rapamycin, Chloroquine (CQ) and 3-methyladenine (3-MA). The effects of different stages of autophagy on viral replication were detected by Western blotting and plaque assay.@*Results@#The study showed that EV71 infection could induce incomplete autophagy in RD cells. Replication of EV71 was promoted in the CQ treatment group, while the level of replication of the rapamycin and 3-MA treatment groups was reduced. Further studies have shown that mTOR is a key molecule affecting EV71 replication, and inhibition of mTOR by rapamycin can inhibit the synthesis of viral RNA.@*Conclusions@#The effects of different stages of autophagy on viral replication are also different. Inhibition of autophagic lysosome degradation promotes viral replication, while inhibition of the early stage of autophagy or promotion of autophagy reduces the level of viral replication. mTOR can affect the replication of EV71 at RNA level.

5.
Article in Chinese | WPRIM | ID: wpr-711382

ABSTRACT

Type Ⅶ secretion system(T7SS) is a novel and specialized secretion system discovered in recent years. It was first found in Mycobacterium tuberculosis. Type Ⅶ secretion system is involved in the secretion of virulence-associated proteins, the interaction between pathogens and hosts and the balance of zinc/iron ions. Moreover,it plays a critical role in the growth and pathogenesis of Mycobacteria. This review summarizes the components,substrates and translocation mechanisms of the type Ⅶ secretion system and its relation to the virulence of Mycobacteria aiming to provide references for developing novel strategies for disea-ses diagnosis,treatment and prevention.

6.
Article in Chinese | WPRIM | ID: wpr-806659

ABSTRACT

The family Paramyxoviridae is a group of viruses that significantly affect human and animal health. Paramyxoviruses are generally thought to enter host cells by direct fusion of the viral and host cell membranes. Membrane fusion, essential for syncytium formation, is not only a pathological hallmark of paramyxoviral infections, but also a kind of mechanism of cell-to-cell viral spread. Nevertheless, the process of membrane fusion is highly conserved, only consisting of two-component fusion apparatus: an attachment protein (HN/G/H) and a fusion (F) protein. However, there is a significant knowledge gap on the mechanism(s) by which HN/H/G couples receptor binding to F-triggering, since different attachment proteins possess special triggering process. In this review, we summarize the general property and distinction of the fusion process during paramyxoviral infection for further research on the pathogenic mechanism.

7.
Article in Chinese | WPRIM | ID: wpr-806639

ABSTRACT

Objective@#BG-derived HIV-1 Tat protein from an HIV-associated dementia (HAD) patient was expressed in E. coli BL21(DE3) and purified in order to research the effects on human umbilical vein endothelial cells (HUVECs) activity.@*Methods@#The recombinant plasmid pGEX-KG-tat with HIV-1 tat stored in our laboratory was amplified by PCR. The PCR product was cloned into pET-32a-tat. The recombinant plasmid pET-32a-tat was transfected into E. coli, and Tat protein was expressed in BL21(DE3), which was induced by IPTG. Then it was purified by Ni-chelating chromatography column and gel filtration preloaded column, and identified by SDS-PAGE and Western blot(WB). The concentration was determined by BCA Kit. Different concentrations of Tat were added into HUVECs to detect their effects on cell activity by cck-8.@*Results@#The Tat with high purity was efficiently expressed in BL21 (DE3) and obtained by using the Ni-chelating chromatography column and gel filtration preloaded column. The concentration was 0.47 mg/ml by using BCA Kit. As the concentration of Tat increased, HUVECs activity decreased. There was no significant difference in cells viability between negative control with 100 ng/ml and 200 ng/ml group (P>0.05). There was significant difference in cells viability between negative control with 300 ng/ml, 400 ng/ml, 500 ng/ml and 1000 ng/ml group (P<0.05). But the difference between 300 ng/ml, 400 ng/ml, 500 ng/ml and 1000 ng/ml group was not statistically significant (P>0.05).@*Conclusions@#The HIV-1 Tat with biological activity was efficiently expressed in BL21 (DE3), and the activity of HUVECs was significantly decreased when the concentration reached 300 ng/ml.

8.
Article in Chinese | WPRIM | ID: wpr-806504

ABSTRACT

Objective@#To identify the function of 91-112 amino acids (aa) fragment, the interaction domain of head and stalk of Newcastle disease virus(NDV) HN glycoprotein, and clarify the role of the fragment in promoting cell specific membrane fusion.@*Methods@#The specific gene sequences were identified by aligning 91-112 amino acids of NDV HN protein with amino acids of MeV H, RSV G, hPIV3 HN protein. The fragment deletion, fragment substitution and intermolecular homologous recombination method were combined to construct the deletion mutant, De(HN), and three chimeras, Ch(MeV), Ch(RSV), Ch(hPIV3). Cationic transfection reagent was used to transfect the plasmids into baby hamster kidney cells (BHK-21), in which vaccinia virus-T7 RNA polymerase expression system was expressed. Indirect immunofluorescence assay (IIFA) and flow cytometry (FCM) were executed to analyze the cell surface expression level. Cell fusion promotion activity, receptor recognition activity and neuraminidase activity of each mutant were also detected.@*Results@#Cell surface expression efficiency of De(HN) and Ch(MeV), Ch(RSV), Ch(hPIV3) proteins were 9.04%, 82.20%, 70.16%, 75.65% of that of wild-type (wt) HN. Fusion promotion activity of De(HN), Ch(MeV), Ch(RSV), Ch(hPIV3) were 3.83%, 24.76%, 29.42%, 57.84% of that of wt HN. The fusion promotion activity of De(HN) almost disappeared and syncytium couldn’t be found under the microscope. Hemadsorption activity was 13.48%, 36.25%, 34.93%, 65.22%, respectively (P<0.05), which was consistent with the fusion promotion activity of mutant proteins. Neuraminidase activity was 10.81%, 54.42%, 50.13%, 60.35% of that of wt HN, respectively (P<0.05).@*Conclusions@#The amino acids fragment (91-112) of NDV HN protein plays an important role in promotion fusion. The loss of fusion promotion activity of De(HN) protein was related to the failure of effective cell surface expression of the mutant.

9.
Article in Chinese | WPRIM | ID: wpr-805921

ABSTRACT

Oncolytic virus therapy is becoming a new direction for cancer treatment, which could take the advantage of the characteristic of oncolytic virus selectively replicating in cancer cells, and killing tumor cells without damaging normal cells. Compared with conventional chemotherapy or radiotherapy, it has higher specificity, fewer side effects and the ability resisting various kinds of malignant tumors. Newcastle disease virus, a typical oncolytic virus, can cause Newcastle disease in poultry. However, no serious symptoms occurred after human being infected with NDV. With the development of reverse genetics technology, it is possible to enhance the anti-tumor activity of NDV by promoting membrane fusion and apoptosis with gene recombination. The review is about the recent research progress in vitro and in vivo oncolytic experiments and clinical application of NDV at home and abroad, which aimed at providing scientific reference for the anti-tumor study of NDV in the future.

10.
Article in Chinese | WPRIM | ID: wpr-805908

ABSTRACT

Objective@#To study the sequence characteristics and variation of HIV-1 Vpr gene in different parts of an AIDS dementia complex (ADC) patient and provide basis for the study of the neurologic pathogenesis of HIV-1-assciatd dementia.@*Methods@#Genomic DNA was extracted from peripheral samples (lymph nodes, spleen, liver) and central nervous system (meninges, frontal lobe, temporal lobe gray matter, frontal white matter, basal ganglia cortex) of an ADC patient, The Vpr gene was amplified with nested polymerase chain reaction (PCR). PCR products were cloned into the pMD19-T vector. After transformation into DH5α competent E. coli, five positive clones were sequenced. The phylogenetic tree was built and genetic distance was calculated through MEGA6, and the values of ds/dn was calculated through SNAP, then the changes of the amino acid sites were analyzed.@*Results@#HIV-1 Vpr genes isolated from different tissues of the ADC patient had variations. Vpr HIV-1 gene sequences from central nervous system and peripheral tissues were intercrossed together in the phylogenetic tree. Central nervous system and peripheral HXB2 Vpr had no significant differences in genetic distance. The ds/dn of all the HIV-1 Vpr gene sequences were 3.3749.@*Conclusions@#The HIV-1 Vpr sequences were different in the ADC patient, and there were different variations in different parts of the peripheral and central regions. Whether these variations are related to the pathogenesis of ADC remains to be further studied.

11.
Article in Chinese | WPRIM | ID: wpr-612726

ABSTRACT

Objective To study the clinical evaluation of the effect of topical application of Periocline ointment in the adjuvant treatment of periodontitis.Methods100 patients with periodontitis as the research object in this study in Xi'an Jiaotong university school hospital from April 2015 to August 2016 were randomly divided into the control group and the experimental group, 50 cases in each group.The control group were treated with the conventional treatment, at this basis, the experimental group were given Periocline ointment topical application as the auxiliary treatment.Eight weeks after treatment, the clinical treatment and related indicators were compared in the two groups.ResultsThe bleeding index, probing depth and attachment level in the experimental group were significantly better than those in the control group (P<0.05).In the experimental group of 50 patients, the total effective number of cases in the treatment of 48 cases, The total effective rate in the experimental group was 94.0%, which was significantly higher than 88.0% in the control group (P<0.05).ConclusionIt can improve the effective rate that Periocline ointment was local applicated in the adjuvant treatment of periodontitis;it is worthy of clinical application.

12.
Chinese Journal of Nephrology ; (12): 495-503, 2017.
Article in Chinese | WPRIM | ID: wpr-611044

ABSTRACT

Objective Intradialytic hypotension (IDH) is one of the common complications during hemodialysis,however its diagnostic criteria are highly controversial at present.In order to fully understand the prevalence of IDH in our center and figure out which diagnostic criteria is better for Chinese maintenance hemodialysis (MHD) patients,we choose several IDH definitions by reviewing published literatures and analyze their association with mortality.Methods The patients were recruited from Blood Purification Center of Ruijin Hospital undergoing hemodialysis during July 2012.Pre-,intra-and post-dialysis blood pressure were recorded.Patients' clinical characteristics,laboratory results and cardiac ultrasound results were collected.Based on several IDH definitions,we investigated the prevalence rate of IDH and its frequency among MHD patients.SPSS 23.0 was used to analyze data and conduct survival analysis.Results Totally 219 MHD patients underwent 16084 hemodialysis in 6 months.The prevalence rate,overall and individual frequency of IDH fluctuated between 45.21%-100.00%,4.64%-37.60% and 0.00%-33.00% respectively.For every IDH criteria,the patients were recruited into the group IDH(+) if they ever met the corresponding definition,otherwise the group IDH(-).Survival analysis found that IDH (the criteria of an absolute systolic blood pressure (SBP) < 90 mmHg or with a decrease of SBP≥ 20 mmHg) could decrease the risk of patients' cardiovascular mortality but was not relevant to all-cause mortality.Further analysis showed these patients had better cardiac functions mainly reflecting in lower Pro-BNP (2880 ng/L vs 6909 ng/L),lower prevalence rate of left ventricular hypertrophy (52% vs 83%) and higher left ventricular ejection fraction (65.0% vs 62.5%) than IDH(-) patients.No correlation was found between other IDH criteria and mortality.Conclusions The prevalence rate,overall and individual IDH frequency of IDH are of high variability when diagnosed by different IDH criteria.All IDH episodes defined by our selected definitions are of no association with all-cause mortality.An absolute SBP < 90 mmHg or with a decrease of SBP≥20 mmHg can decrease the risk of cardiovascular mortality due to their better cardiac function.Large scale researches should be conducted to find optimal IDH definition and explore the association of IDH and mortality.

13.
Article in Chinese | WPRIM | ID: wpr-618734

ABSTRACT

Objective To compare the degree of cell injury induced by 3D protein (SDLY11 and SDLY107) of enterovirus 71 (EV71) strains.Methods EV71 strains SDLY11 and SDLY107 were respectively isolated from children with mild and severe hand foot mouth disease.The target genes 11-3D-Flag and 107-3D-Flag were amplified by reverse transcription-polymerase chain reation (RT-PCR) and inserted into the eukaryotic expression vector pcDNA3.1.The recombinant plasmids 11-3D-Flag-pcDNA3.1 and 107-3D-Flag-pcDNA3.1 were transformed into Escherichia.coli DH5α, respectively, and were identified by enzyme digestion and sequencing.The recombinant plasmids were transfected into rhabdomyosarcoma (RD) cells, respectively.Expression of 3D protein was detected by indirect immunofluorescence assay and western blot.Cell injury induced by 3D protein was detected with lactate dehydrogenase (LDH) test, cell proliferation was detected by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenylthiazolium bromide (MTT) test, and cell apoptosis was detected with Annexin-V and PI.Multiple comparisons among groups were analyzed using LSD-t test if multiple sets of variables were consistent with homogeneity of variance.If not, Dunnett T3 test was used.Results The 1 400 bp fragments were amplified by reverse tramscription (RT)-polymerase chain reaction (PCR), and the recombinant plasmids were digested by enzyme and the 1 400 bp and 5 400 bp fragments were obtained and identified.Gene sequencing showed that the sequences were consistent with the target genes.The specific fluorescence was observed by indirect immunofluorescence assay, and the western blot showed that the molecular weight of the target protein was 55×103.The LDH test showed that the A490 of SDLY11 3D protein transfection group (0.790±0.048) was higher than that of SDLY107 3D protein transfection group (0.641±0.018).The difference was statistically significant (t=5.14, P<0.05).The cell membrane damage caused by SDLY11 3D protein was more severe than SDLY107 3D protein.The MTT test showed that the A570 of SDLY11 3D protein transfection group (1.028±0.020) was lower than that of SDLY107 3D protein transfection group (1.081±0.002), and the difference was statistically significant (t=3.31, P<0.05).The effect on cell proliferation activity of SDLY11 3D protein was greater than SDLY107 3D protein.The results of Annexin-V/PI showed that the percentage of apoptotic cells of SDLY11 3D protein transfection group and SDLY107 3D protein transfection group were (1.471±0.246)% and (1.465±0.237)%, respectively, and the difference was not statistically significant (t=0.04, P=0.973).Conclusions Compared with the SDLY11 3D protein, SDLY107 3D protein induces slighter cell injury, has weaker effect on cell proliferation activity, and is more favorable for virus replication in cells.

14.
Recent Advances in Ophthalmology ; (6): 776-779,784, 2017.
Article in Chinese | WPRIM | ID: wpr-609874

ABSTRACT

Objective To evaluate the clinical efficacy of intravitreal injection of ranibizumab combined with panretinal photocoagulation for severe non-proliferative diabetic retinopathy.Methods Sixty-four cases (90 eyes) with severe non-proliferative diabetic retinopathy from July 2014 to June 2016 in our hospital were randomly divided into two groups according to the digital table.The observation group (32 cases,46 eyes) underwent intravitreal injection of reizumab combined with panretinal photocoagulation,and the control group (32 cases,44 eyes) were treated with only panretinal photocoagulation.The visual acuity,fundus,fundus fluorescein angiography and macnlar thickness postoperative 1 month,3 months,6 months were observed and detected.The mean thickness of the retinal neuroepithelium in the macular region and total volume of the 6 mm diameter neuroepithelium in the macular area were compared and statistically analyzed.The laser energy,number of spots and energy density were calculated and recorded.Results BCVA at postoperative 1 month,3 months,6 months in the observation and the control group were better than that before treatment(all P < 0.05),and the observation group was better than the control group (all P < 0.05).In the control group,BCVA had no statistically significant difference between 3 months and 6 months (P > 0.05),and the differences were statistically significant among other time points (all P < 0.05).In the observation group,the differences were statistically significant among all time points after treatment (all P < O.05).In the comparison of mean thickness of the retinal neuroepithelium in the macular region and total volume of the 6 mm diameter neuroepithelium in the macular area in the control group and the observation group before and after treatment,there were statistically significant differences at different time points between the two groups (all P < 0.05).Moreover,the average thickness of the retinal neuroepithelium and total volume of 6 mm diameter neuroepithelium in the macular region at postoperative 1 month,3 months,6 months were better than those in the control group (all P < 0.05).The laser energy,spot number and energy density of the observation group were significantly lower than those of the control group (all P < 0.05).Conclusion Intravitreal injection of ranibizumab combined with panretinal photocoagulation for severe non-proliferative diabetic retinopathy can accelerate retinal neovascularization in a short period of time,reduce macular edema and improve the visual function of patients with less laser energy,the curative effect is better than application of retinal laser photocoagulation alone,is worthy of clinical promotion.

15.
Article in Chinese | WPRIM | ID: wpr-808662

ABSTRACT

Human parainfluenza viruses (hPIVs), a series of single-stranded RNA viruses of Paramyxoviridae, are the main pathogen of respiratory tract infection. hPIV3 is the main cause of lower respiratory tract infection leading to bronchiolitis and pneumonias in young children under the age of six months, and it is the second major pathogen only next to respiratory syncytial virus (RSV). In this paper, we mainly discuss two kinds of virulence-related surface glycoprotein of hPIV3: hemagglutinin-neuraminidase (HN) protein and fusion protein (F) by briefly introducing the protein structure and physiological functions of HN and F. According to the latest research progress, we focus on the models which have been postulated to explain how F and HN work in concert to bring about membrane fusion.

16.
Practical Oncology Journal ; (6): 527-532, 2016.
Article in Chinese | WPRIM | ID: wpr-506753

ABSTRACT

Objective To explore the efficacy of recombinant human endostatin ( Rh-endostatin ) com-bined with vinorelbine -cisplatin( NP) regimen for newly diagnosed patients with advanced non -small cell lung cancer(NSCLC)with bone metastases and the impact on expression of serum vascular endothelial growth factor (VEGF).Methods From January 1,2009 to February 1,2012,a total of 40 with newly diagnosed,advanced NSCLC patients with bone metastases were enrolled in this study and randomly assigned to the treatment (N=20) or control(N=20)group.The control group was only given the NP regimen chemotherapy and the treatment group was treated with Rh-endostatin combined with NP regimen .The changes in clinical effects of the two groups and serum VEGF levels were observed .Results After two cycles of systemic chemotherapy ,objective response rate (ORR)and disease control rate(DCR)of the treatment group were 30.0% and 80.0%significantly higher than 5.0%and 45.0%of the control group(P0.05).Conclusion Rh-endostatin combined with NP regimen can im-prove the efficiency of treatment for newly diagnosed patients with advanced NSCLC with bone metastases and does not increase adverse reactions whereas no significant difference in serum VEGF Levels .

17.
Article in Chinese | WPRIM | ID: wpr-506235

ABSTRACT

Objective:To study the occlusion characteristics of the young subjects with bruxism before and after treatment by elastic occlusal splint. Methods:10 patients with bruxism and aged 22-27 years(n=10) were treated by elastic occlusal splint, before and 6 months after treatment they were examined by T-Scan occlusion detector. The occlusion time( OT) , disclusion time( DT) , oc-clusion force( OF) , the perentage of occlusion force( POF) , the changes of the position of center occlusal force( COF) and the asym-metry index of occlusion force(AOF) were observed and compared with those of the healthy controls(n=10). Results: ① In the case group there was significant difference in the OT and DT between left and right anterior teeth before treatment (P0. 05). There was significant difference in the momevent of COF of left and right side, the POF of right side and the AOF between the 2 groups before treatment, but not after the treatment(P>0. 05). ② In the case group after treatment the OT and DT of left and right anterior teeth were shortened(P0. 05). Conclusion:The occlusal factors such as early contact, lateral teeth synthetic interference, occlusal force center excursion have close relationship with bruxism. Elastic occlusal splint can effectively correct muscle dysfunction and make occlusal relationship more coordinated and stable, and therefore is effective in the treatment of bruxism.

18.
Practical Oncology Journal ; (6): 167-171, 2016.
Article in Chinese | WPRIM | ID: wpr-499275

ABSTRACT

Immunosuppressive tumor microenvironment is a part of the tumor microenvironment that plays an immunosuppressive function.It consists of suppressor cells and inhibitory cytokines.In recent years,be-cause of its important role in tumor immunity,CAR-T and PD-1/PD-L1 signal pathway has become a hot top-ic of immunotherapy, which can suppress immune through different mechanisms to promote tumor progression. Therefore,taking more effective anti-tumor therapies against the above mechanisms is possible to rescue the pro-gress of tumor.In this article,the influence of immunosuppression microenvironment in CAR-T cell therapy and PD-1/PD-L1 signaling pathway in the anti-tumor process is reviewed.

19.
Chinese Journal of Virology ; (6): 468-473, 2015.
Article in Chinese | WPRIM | ID: wpr-296261

ABSTRACT

Enterovirus A71 (EV-A71) causes hand, foot, and mouth disease (HFMD) and various neurological complications, including aseptic meningitis and neurogenic pulmonary edema in young children. HFMD caused by EV-A71 have broken out several times in the Asia-Pacific region since 2007. And it has been a serious threat to public health. There is no effective vaccine or antiviral drug. The pathogenesis of EV-A71 infection is unknown, and EV-A71 3C protein plays an irreplaceable role in replication and anti - innate immunity. Further research on EV-A71 3C protein is conducive to understand the pathogenesis of EV-A71 infection and antiviral drug.


Subject(s)
Animals , Antiviral Agents , Pharmacology , Enterovirus , Allergy and Immunology , Metabolism , Physiology , Humans , Immunity, Innate , Viral Proteins , Chemistry , Genetics , Metabolism , Virus Replication
20.
Article in Chinese | WPRIM | ID: wpr-672125

ABSTRACT

Objective To construct a chimeric infectious clone of the fatal virulent strain SDLY 107, containing the gene fragments encoding 2A and 3B proteins of the mild virulent strain SDLY 1, and to establish a reverse genetic system platform for further investigation on virulence of enterovirus 71 strains. Methods The overlap PCR analysis was performed to obtain the gene fragments encoding 2A and 3B pro-teins of the mild virulent strain SDLY 1.The obtained gene fragments were digested and then cloned into a plasmid pMD19-T containing the full-length gene of SDLY 107 strain by using gene replacement strategy. The recombinant RNA was transfected into Vero cells for the preparation of recombinant virus particles.Sev-eral assays including the PCR, indirect immunofluorescence ( IFA) , Western blot and sequencing were per-formed for virus identification.Virus titers were measured by 50%cell culture infective dose ( CCID50 ) and plaque assay.Results The infectious clones of SDLY 107-2A-1 and SDLY 107-3B-1 chimeric virus strains were constructed successfully.Typical cytopathic effect was observed in Vero cells after viral transfection. Identification of the rescued viruses by PCR, IFA, Western blot and sequencing further confirmed the suc-cessful construction of infectious virus strains.The virus titers of SDLY 107-2A-1 and SDLY 107-3B-1 strains detected by CCID50 and plaque assay were 1.25 ×105 PFU/ml and 0.7 ×105 PFU/ml, respectively. Conclusion The chimeric viruses SDLY 107-2A-1 and SDLY 107-3B-1 were rescued successfully, causing cytopathic effects similar to those by using the parental virus strain SDLY 107.This study might pave the way for further investigation on in vitro and in vivo virulence of enterovirus 71 strains.

SELECTION OF CITATIONS
SEARCH DETAIL