Chikungunya virus ; genetics ; isolation & purification ; China ; Genome, Viral ; RNA, Viral ; genetics ; Sequence Analysis, RNA

Objective:To explore the changing tendency of IgG to coronavirus associated severe acute respiratory syndrome(SARS).Methods:Between 1 and 153 days after the onset of symptoms, the antibody IgG to SARS associated coronavirus were detected in 534 patients with SRAS by ELISA. 216 randomly selected people without SARS contact history and SARS symptoms were contrasted.Results:The positive rates of IgG between SARS patients and control group respectivel were 58 1%,0 5%.There is a statistical significance(P=0 000).Among SARS patients, the positive rates of IgG in 1～10 days,11～30 days,31～60 days,61～90 days,91～120 days,above 120 days and after the onset of symptoms respectively were 18 8%(3/16 cases),63 8%(37/58 cases),55 6%(30/54 cases),51 4%(71/138 cases),62 1%(159/256 cases),83 3%(10/12 cases).Conclusion:There are low positive rates of IgG in SARS patients in the first 10 days after onset of symptoms, and abruptly ascending positive rates of IgG in 11～30 days.The positive rates of IgG remains the high levels during 4th and 5th months.Further observation is essential to elucidating the duration of IgG.

Background Clinical effectiveness of intavitreal injection of glucocorticosteroid for macular edema has been verified,especially triamcinolone acetonide(TA).However,the efficacy and safety of combination of TA with macular laser grid photocoagulation for macular edema is concerned. Objective This clinical trial was to evaluate the efficacy and safety of intravitreal injection of TA combined with macular laser grid photocoagulation in the treatment of macular edema. Methods A case-cohort study was designed.One hundred and twenty eyes of 120 patients with macular edema from diabetes or retinal vein occlusion were included in this study.The patients were randomized into trial group and control group,with the matched age,course,visual acuity,intraocular pressure (IOP).The patients of the trial group received intravitreal injection of TA combined with macular laser grid photocoagulation,and those of the control group were managed with macular laser grid photocoagulation only.Best corrected visual acuity ( BCVA),optical coherence tomography(OCT),fundus fluorescein angiography(FFA) and IOP were examined before TA injection and 1 week,1 month,3 months,6 months after treatment and compared among different time points between two groups.Written informed consent was obtained from each patient prior to entering this trial. Results Compared with TA injection before,the BCVA was significantly elevated in the trial group 1 week,1 month,3 months and 6 months after TA injection( all P=0.000),however,no obvious improvement of visual acuity was found in the control group before and after treatment at any time point (P＞0.05 ).At various time points,the visual acuity was significantly improved in the trial group than the control group (P =0.037,0.000,0.002,0.046 ).Macular thickness was significantly decreased at various time points after TA injection in comparison with before TA injection in the trial group(all P=0.000),but no significant change in macular thickness in the control group between before and after treatment at any time point( P＞0.05 ).Macular thickness was lower in the trial group compared with the control group at various time points after treatment ( P＜0.05 ).Recurrence of macular edema was seen in 7 eyes ( 1 1.67％ ) 4-6 months,and the IOP raise( ＞21 mmHg)was found in 11 eyes( 14.1％ )after TA injection in the trial group.Conclusions Intravitreal injection of TA combined with macular laser grid photocoagulation can be an effective method in the treatment of macular edema.However,recurrence of macular edema or increase of IOP may occur in a few patients within 6 months after TA injection.A long-term follow-up should be performed for the evaluation of efficacy and safety after intravitreous injection of TA.

Dengue virus (DENV) envelope [E] protein is the major surface protein of the virions that indued neutralizing antibodies. The domain III of envelope protein (EDIII) is an immunogenic region that holds potential for the development of vaccines; however, the epitopes of DENV EDIII, especially neutralizing B-cell linear epitopes, have not been comprehensively mapped. We mapped neutralizing B-cell linear epitopes on DENV-1 EDIII using 27 monoclonal antibodies against DENV-1 EDIII proteins from mice immunized with the DENV-1 EDIII. Epitope recognition analysis was performed using two set of sequential overlapping peptides (16m and 12m) that spanned the entire EDIII protein from DENV-1, respectively. This strategy identified a DENV-1 type- specific and a group-specific neutralizing epitope, which were highly conserved among isolates of DENV-1 and the four DENV serotypes and located at two regions from DENV-1 E, namely amino acid residues 309-320 and 381-392(aa 309-320 and 381-392), respectively. aa310 -319(310KEVAETQHGT319)was similar among the four DENV serotypes and contact residues on aa 309 -320 from E protein were defined and found that substitution of residues E309 , V312, A313 and V320 in DENV-2, -3, -4 isolates were antigenically silent. We also identified a DENV-1 type-specific strain-restricted neutralizing epitope, which was located at the region from DENV-1 E, namely amino acid residues 329-348 . These novel type- and group-specific B-cell epitopes of DENV EDIII may aid help us elucidate the dengue pathogenesis and accelerate vaccine design.
Amino Acid Sequence ; Animals ; Antibodies, Neutralizing ; immunology ; Dengue ; virology ; Dengue Virus ; chemistry ; genetics ; immunology ; Epitope Mapping ; Epitopes, B-Lymphocyte ; chemistry ; genetics ; immunology ; Humans ; Mice ; Molecular Sequence Data ; Viral Envelope Proteins ; chemistry ; genetics ; immunology

Objective To sequence and analyze the envelope (E) gene of type Ⅰ dengue virus isolated from Guangzhou in 2009 for tracing the infection source. Methods The serum samples were collected from patients diagnosed with dengue fever in Guangzhou area during 2009. Dengue virus was isolated and cultured in C6/36 cells.The whole length of E gene was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) and then sequenced. The phylogenetic tree was drawn by neighbor-joining method. The bioinformatics analysis was performed by combining the phylogenetic information and the epidemiology data. Results Four strains of type Ⅰ dengue virus were isolated from 19 samples. E gene of these strains was amplified and sequenced. The phylogenetic analysis showed that 09/GZ/9104 strain and 09/GZ/9236 strain had identical nucleotide sequence and fell within the American/African group, 09/GZ/11534 stain and 09/GZ/11562 strain had similar sequence homology and fell within the Asian group. Conclusion The typeⅠdengue viruses in Guangzhou area in 2009 are imported, which belong to two genotypes and may come from two independent origins respectively.

In this study ,we established Cross Priming Amplification (CPA ) technology for detection of influenza A virus (H1N1) approach ,and evaluated the method through clinical specimens .A set of specific primers were designed for CPA ac‐cording to the conservative gene sequences ,designed and realized in the same temperature reverse transcription of RNA and DNA amplification . The amplification products can be totally enclosed nucleic acid detection device for testing . Fourteen healthy pharyngeal swab specimens ,seven other respiratory viruses ,and six arboviruses strains were used as the controls .We used a method that application of gradient dilution to the H 1N1 virus strain as the control to test the sensitivity of the CPA .We also used 102 clinical pharyngeal swab specimens of H1N1 patients for detection object to evaluate the feasibility of CPA clinical detection .Results showed that the CPA reaction did not appear cross reaction on health cases samples and other viruses .The sensitivity of the CPA was approximately 10 copies/uL in the established method that exactly titer H1N1 virus strain gradient dilution test .As to the positive results among the clinical pharyngeal swab samples collected from patients at different stages after onset ,the CPA had the highest positive detection rate during the first three days after onset (100% ) .While the detection rate from day 4 to day 6 after onset was 79 .31% .After 7 days ,the detection rate was 9 .09% .The established CPA assay was a highly sensitive ,specific and reproducible approach for rapid detection of H1N1 virus ,which is conducive to the early diagno‐sis of influenza A virus (H1N1) for basic medical units .

Objective To investigate the predisposing factors and clinical features of disseminated herpes zoster, and to explore factors influencing postherpetic neuralgia. Methods Clinical data were collected from 53 patients with disseminated herpes zoster and 809 patients with common herpes zoster between 2012 and 2015, and analyzed retrospectively. Logistic regression analysis was used to assess factors influencing the occurrence of and pain intensity in disseminated herpes zoster, as well as the occurrence of postherpetic neuralgia. Results No significant difference in patients′age was observed between the disseminated and common herpes zoster groups(56.66 ± 17.24 vs. 56.50 ± 15.51 years, t=0.071, P>0.05), but there was a significant difference in the gender ratio between the two groups(χ2 = 8.16, P = 0.004). The incidence rates of bullae, pustules and fever were all significantly higher in the disseminated herpes zoster group than in the common herpes zoster group(15.09%vs. 3.58%,χ2=16.04, P<0.01;47.17%vs. 26.82%,χ2=10.20, P<0.01;30.19%vs. 8.03%,χ2=28.68, P<0.01). The disseminated herpes zoster group also showed significantly higher pain scores at admission compared with the common herpes zoster group (Median[P25- P75]: 6[4- 7.5] vs. 5[3- 7], Z =-3.460, P = 0.001). Logistic regression analysis revealed that gender, age, fatigue and HIV infection were significantly associated with the occurrence of disseminated herpes zoster (all P<0.05). Additionally, HIV infection(OR=5.570, 95%CI:1.196-25.939, P=0.029), gender(OR=0.166, 95%CI:0.029-0.945, P=0.043), age(OR=1.064, 95%CI:1.010-1.119, P=0.019)and the number of days that antiviral therapy lasted(OR=0.669, 95%CI:0.505-0.885, P=0.005)were all factors influencing the occurrence of postherpetic neuralgia. Conclusion Male, old age, fatigue and especially HIV infection are risk factors for the occurrence of disseminated herpes zoster, and male, old age and antiviral therapy duration may be associated with the occurrence of postherpetic neuralgia.

Facing current situation of integrative medicine (IM), authors put forward that clinical and diagnosis program of IM could be carried out from clinical path, pathogenesis, treatment theory and philosophy, and so on, but with different integration degrees. Meanwhile, formulation of concrete program should be disease-targetedly set up, and adjusted from person to person, from place to place, from time to time. As for settled IM program , authors could evaluate it from whether Chinese medicine and Western medicine have formed complementary, synergistic, excitatory actions, and toxicity attenuation; whether more problems could be solved in efficacy, safety, practicability, and economy than previous single mode.
Critical Pathways ; Integrative Medicine ; trends ; Medicine, Chinese Traditional ; trends

Objective To construct a recombinant expression vector for expression of the function-al domains of dengue virus serotype 1 ( DENV1 ) envelope ( E ) protein in native soluble form. Methods The genes encoding the functional domains of DENV1-E protein (1-394 aa) were amplified with PCR and then cloned into the Psectag2B-Fc eukaryotic expression vector.The 293T cells were transfected with the recombinant vector by cationic lipid-based delivery.The cell clones expressing the fusion DENV1-E-Fc protein were screened out with 2 mg/ml of Zeocin.Immunofluorescence assay ( IFA) was performed to analyze the antigenicity and integrity of the fusion protein.The fusion proteins were purified from cell lysate with Protein-G and further identified by Western blot assay.Results The soluble form of fusion protein with a molecular weight of about 90×103 was obtained at a yield of about 25 μg per 1×107 cells.The results of IFA indicated that the fusion protein kept its integrity with right conformational epitopes.The fusion protein was successfully expressed with the advantage of good specificity as indicated by IFA and Western blot assay. Conclusion The recombinant fusion protein in soluble form was successfully expressed in eukaryotic ex-pression system, which paved the way for further investigation on the function of DENV1 E protein and its protective epitopes.

ObjectiveTo analyze the Envelope (E) gene of type 1,2,3 dengue virus isolated fromGuangzhouin2010, andtoinvestigatetheinfectionsourceandvirusgenotypes.MethodsEighty-five serum samples were collected from 85 patients in acute phase of dengue fever.Dengue virus was cultured and isolated by C6/36 cells.The whole length of E gene was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) and then sequenced.The phylogenetic tree was drawn by neighbor-joining method.The bioinformatics analysis was performed by combining the phylogenetic information and the epidemiologic data.ResultsSix strains of type 1 dengue virus,two strains of type 2 dengue virus and six strains of type 3 dengue virus were isolated from 85 samples.The E gene sequence of these strains was obtained by sequencing.The phylogenetic analysis showed that type 1 and 3 dengue virus belonged to two genotypes (Asian and South Pacific ocean,India subcontinent and Southeast Asia/South Pacific ocean,respectively),and type 2 dengue virus belonged to one genotype (Malaysia/India subcontinent).ConclusionIt's presumed that all strains of type 2 dengue virus are imported,four strains of type 1 dengue virus are imported and four strains of type 3 dengue virus arc imported,the remaining two stains of type 1 and two stains of type 3 dengue virus need mosquito intermediary research further to prove their origins.