Venezuelan equine encephalitis (VEE) is a zoonotic disease caused by the Venezuelan equine encephalitis virus (VEEV) complex. This disease has not yet been reported in China, and it is therefore essential to establish a rapid and accurate method for detection of the virus in order to prevent and control this disease. In this study, a one-step real-time quantitative RT-PCR method was developed for the detection of the VEEV complex. A pair of specific primers and a Taqman probe were designed corresponding to a conserved region of the VEEV gene nspl, allowing the detection of all known strains of different sub- types of the virus. Using RNA synthesized by in vitro transcription as template, the sensitivity of this method was measured at 3.27 x 10(2) copies/microL. No signal was generated in response to RNA from Chikungunya virus (CHIKV), nor to RNA encoding the nsp1 fragment of Eastern equine encephalitis virus (EE-EV) or Western equine encephalitis virus (WEEV), all of which belong to the same genus as VEEV. This indicates that the method has excellent specificity. These results show that this one-step real-time quantitative RT-PCR method may provide an effective tool for the detection of VEEV in China.
China ; DNA Primers ; genetics ; Encephalitis Virus, Venezuelan Equine ; classification ; genetics ; isolation & purification ; Encephalomyelitis, Venezuelan Equine ; virology ; Humans ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods

The status of acupuncture-moxibustion is more and more recognized by mainstream medicine in the world in recent years, and literature regarding acupuncture-moxibustion with high impact factor (IF) published in the worldwide mainstream medicine journals is also gradually growing by years. To understand the situation of related literature, literature regarding acupuncture-moxibustion with IF of more than 10 in Science Citation Index (SCI) during the recent 5 years was retrieved. The number, the types, the diseases involved, the publishing states of the acquired articles and the source, the citation, the IF of the publishing journals were analyzed and summarized. Additionally, some of the research foci, the new research tendencies and the deficiencies of research were discussed. The thoughts and suggestions are expected to be provided for further research of acupuncture.
Acupuncture Therapy ; statistics & numerical data ; Bibliometrics ; Humans ; Journal Impact Factor ; Moxibustion ; statistics & numerical data ; Publications ; statistics & numerical data

Objective To quantify the changes of tongue color with the development of the disease according to the characteristics of human vision.Methods Psychological scales of tongue color perception were obtained from thc psychophysical experiments.Polynomial regression and support vector regression (SVR) were used to establish the mathematical model between the physical values and the psychological scales of tongue color.Results The psychological scales exported from the model could correspond to the visual perception of tongue color change, and SVR has higher accuracy.Conclusions The psychological scale, can not only quantitatively evaluate the tongue color in the development of the disease, but also quantify the degree of disease, to assist the doctors for disease diagnosis and treatment.

Objective To characterize the changes of retinal structure and visual field mean defects (MD) in early Parkinson's disease (PD) and the related factors.Methods Twenty-five patients with early stage PD and 33 normal controls were enrolled in this study.PD patients were evaluated by unified Parkinson's disease rating scale (UPDRS) and Hoehn-Yahr (H-Y) stage in off period.All subjects undertook retina of retinal nerve fiber layer (RNFL) and macular examinations by optical coherence tomography (OCT),and MD by automatic visual field analyzer.Results (1) The average thicknesses of RNFL and C11 quadrant RNFL were thinner in PD group [(96.2 ±7.6) μm and (124.4 ± 18.4) μm] than in healthy controls [(102.6 ± 5.0) μm and (135.4 ± 21.8) μm,respectively,P =0.000 and P =0.047].Moreover,PD patients had a thinner average thickness of macular[(277.2 ±9.6) μm vs (285.8 ± 12.6)μm,P=0.006],and smaller macular volume compared with controls [(10.0 ± 0.3) mm3 vs (10.3 ±0.4) mm3,P =0.006].However,there was no significant difference in MD between PD and control subjects [(0.43 ± 2.75) dB vs (-0.18 ± 1.41) dB,P =0.322].(2) Pearson's correlation analysis showed that H-Y stage was negative correlated with the average RNFL thickness (r =-0.569),average macular thickness and volume (r=-0.501 and r=-0.417) in PD patients (all P＜0.05).Conclusions PD patients have thinner average thicknesses of RNFL,C11 quadrant RNFL and macular,smaller macular volume than those in normal controls.There were no significant differences in MD between two groups.There were negative relationships between H-Y stage,and retinal thickness and volume of PD patients.

Objective To compare the sensitivity and specificity of dot immunogold method (DIM) and particle agglutination (PA) for the diagnosis of mycoplasma pneumoniae (MP) infection. Methods The 190 serum specimens of 113 children with mycoplasmal pneumonia (infection group) and 50 serum specimens of 50 health children (health group) were tested for MP by PA and DIM- A and B. Results In infection group, the positive rates of DIM- A and B were 82.63% (157/190) and 84.74%(161/190), and there was no statistical difference (χ2 = 0.31, P>0.05); the positive rate of PA (titer ≥1:160) was 70.00%(133/190), the positive rate of PA was significantly lower than that in DIM-A and B, and there were statistical differences (P<0.05). In infection group, with the increase of PA titer, the positive rate of DIM was gradually increased, and there was a correlation between 2 methods (rA=0.972, rB=0.830);the positive rates of DIM- A and B in serum specimens of PA negative were 40.62%(13/32) and 53.12%(17/32), and in the PA titer of 1:5120, there was still a negative result in DIM. In health group, the positive rates of DIM-A and B were 26.00% (13/50) and 28.00% (14/50), and there was no statistical difference (χ2 = 0.66, P>0.05); the positive of PA was 8.00% (4/50), the positive rate of PA was significantly lower than that in DIM- A and B, and there were statistical differences (P<0.05 or<0.01). Conclusions Compared with the PA, DIM has low sensitivity and poor specificity for clinical diagnosis. DIM is not suitable for clinical diagnosis of MP infection.

To study the chemical constituents of Veratrum dahuricum (Turcz.) Loes. f., a new aurone glycoside named as (Z)-7, 4'-dimethoxy-6-hydroxyl-aurone-4-O-β-glucopyranoside was isolated from the 95% ethanol extracts of the rhizomes and roots of Veratrum dahuricum (Turcz.) Loes. f. by repeated column chromatography on silica gel and recrystallization. Its structure was established by extensive spectroscopic analyses, and its cytotoxicities against HepG-2, MCF7 and A549 cell lines were measured in vitro.

Objective To access the diagnostic accuracy and clinical value of transvaginal real time three-dimensional hysterosalpingo-contrast sonography (RT-3D-HyCoSy) in evaluating the fallopian tube patency.Methods 428 infertile women were under went transvaginal RT-3D-HyCoSy and 32 of them were underwent laparoscopically salpingoraphy to evaluate the fallopian tube patency.Results 846 fallopian tubeswere studied by RT-3D-HyCoSy,271 of them were unobstructed,202 of them were obstructed,373 of them were partially passable.Compared with laparoscopically salpingography,sensitivity,specificity,positive predictive value and negative predictive value of transvaginal RT-3D HyCoSy were 84.0％,100％,87.5％ and 90.9％,respectively.The transvagianl RT-3D HyCoSy and the laparoscopy with a Cohen's Kappa was 0.788.Conclusions Transvagianl RT-3D HyCoSy could dynamic show the visualization of the course from uterus to distal tubal and the both ovary consecutively.It has more advantage in assessing the patency of tubal and distal tubal function.

Objective : To investigate the epidemiological characteristics of heatstroke and its correlation with meteorological factors in Shaoxing in 2017,and to provide evidence for heatstroke prevention and control. Methods : The data of heatstroke cases and the daily meteorological indexes were collected from July 2017 to August 2017 in Shaoxing to describe the spatial,temporal and population distribution of heatstroke cases. The correlation between heatstroke and meteorological factors was analyzed by multivariate logistic regression model. Results : A total of 759 cases of heatstroke were reported,with an average age of（53.3 ±17.9）years. There were 487 males cases（64.16%）and 272 female cases（35.84%）. There were 618 cases of mild heatstroke（81.42%）and 141 cases of severe heatstroke（18.58%）. There were six cases of death from severe heatstroke,and the mortality of severe heatstroke was 4.26%. Minimum temperature（rs=0.851,P<0.001）,maximum temperature（rs=0.726,P<0.001）and wind speed（rs=0.285,P=0.025）were positively correlated with the incidence of heatstroke,and relative humidity（rs=-0.693,P<0.001）and rainfall（rs=-0.414,P=0.001）were negatively correlated with the incidence of heatstroke. The results of logistic regression analysis showed that high daily minimum temperature was a risk factor for severe heatstroke（OR=1.854,95%CI：1.606-2.140）. Conclusion The mortality of severe heatstroke patients was high in Shaoxing,the daily minimum temperature was correlated with severe heatstroke.

Objective:To analyze the molecular characteristics of Echovirus 11 (Echo11) strains isolated in Xiangyang, Hubei Province from 2016 to 2017 based on the sequences of VP1 gene.Methods:Rectal and throat swab specimens were collected from children with hand, foot and mouth disease (HFMD) in Xiangyang from 2016 to 2017. Echo11 strains were detected by real-time reverse transcriptase PCR (RT-PCR) and isolated after cultured in human rhabdosarcoma (RD) cells. The VP1 regions of Echo11 strains isolated from RD cells and the whole genomes of three representative Echo11 strains were amplified by conventional RT-PCR and the sequences were analyzed. DNAStar7.0 (MegAlign) and MEGA6.0 (Data) were used to analyze the homology and mutation sites in nucleotide and amino acid sequences. Neighbor-joining method was used to construct phylogenetic trees. Recombination analysis was performed with SimPlot software (BootScanning).Results:A total of 11 Echo11 strains were isolated from 3 494 HFMD cases, accounting for 0.31%. They were highly homologous in the VP1 gene. These strains shared 98.4%-100.0% homology in nucleotide sequences and 98.3%-100.0% homology in amino acid sequences. The homology between the 11 Echo11 strains and the prototype strain (Echo11/Gregory, X80059) was 73.9%-74.8% in nucleotide sequences and 87.7%-88.7% in amino acid sequences. All of the Echo11 strains circulating in Xiangyang were classified into lineage D, having a similarity to the strains circulating in some regions of mainland China since 2013. In multiple regions of the genome, the Echo11 strains isolated in Xiangyang were highly similar to the Henan Echo1 strains in 2010 and the Hubei Echo6 strains in 2015, suggesting there was recombination within the genome of Echo11 strains in Xiangyang.Conclusions:The Echo11 strains circulating in Xiangyang from 2016 to 2017 belonged to lineage D and were recombinant strains.

Objective To investigate the effect of different concentration of uric acid on proliferation and phentyic change of HBZY-1 cells in vitro. Methods The morphological change of HBZY-1 cells exposed to uric acid was observed under the inverted microscope and transmission electronmicroscope. MTT bioassay was used to assess the effect of uric acid on the proliferation of HBZY-1 cell. ELISA method was used to assess the level of angiotensin Ⅱ in HBZY-1 cell incubated with different concentration of uric acid for different treatment duration.The protein and mRNA expression of α-smooth muscle actin (α-SMA) , transforming growth factor-β1 (TGF-β1) and fibronectin in HBZY-1 cells was measured by Western blot and RT-PCR assay, respectively. Results Uric acid (0.4 mmol/L, 24 h) could induce phentypic change of messangial cell from triangle to fibroblast-like morphology.Uric acid could significantly stimulate the proliferation of HBZY-1 cells for 24 h in a concentration-dependent manner (P < 0.05). Uric acid could stimulate the secration of angiotensin Ⅱ in a dose-and time-dependent manner.RT-PCR and Western blot results showed that the markers of myofibroblast transdifferentiation, including a-smooth muscle actin (α-SMA) , transforming growth factor-β1 (TGF-β1) and fibronectin, were significantly upregulated in HBZY-1 cell treated with over 0.1 mmol/L uric acid for 48 h compared with the control group (P < 0.05, respectively). Conclusion Uric acid can induce the proliferation and phentypic change of HBZY-1 in vitro.