IL-27 regulates NLRP3 inflammasome activation of MDSCs in experimental Sjögren's syndrome.

Excessive NOD-like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome activation has an important function in the pathogenesis of Sjögren's syndrome (SS). Increased and dysfunctional myeloid-derived suppressor cells (MDSCs) promoted SS. However, NLRP3 inflammasome activation of MDSCs in SS and its regulated components are unclear. Splenic MDSCs were purified by immunomagnetic beads and cultured. Western blot was used to assess NLRP3 inflammasomes. Interleukin-1ß (IL-1ß) and IL-18 were measured using enzyme-linked immunosorbent assay. Here we showed that the NLRP3 inflammasome was activated in non-obese diabetic (NOD) mice with SS-like manifestations. We found that NLRP3 inflammasome activation was augmented in MDSCs of SS mice and NLRP3 inflammasome activation was suppressed in IL-27-deficient NOD mice. Consistent with findings of SS mice in vivo, we observed that NLRP3 inflammasome activation by adenosine triphosphate and lipopolysaccharide was remarkably intensified in MDSCs with IL-27 treatment in vitro. Collectively, our data highlighted that IL-27 regulates NLRP3 inflammasome activation of MDSCs in experimental SS.

Total glucosides of paeony alleviates experimental Sjögren's syndrome through inhibiting NLRP3 inflammasome activation of submandibular gland cells.

OBJECTIVES: The mechanisms by which total glucosides of paeony (TGP) mitigates Sjögren's syndrome (SS) remains elusive. In the present study, we aim to explore the relationship between the therapeutic effects of TGP in the treatment of SS and NLRP3 inflammasome activation in submandibular gland (SG) cells. METHODS: Female non-obese diabetic (NOD) mice were selected as the model of SS. The mice were divided into PBS and TGP treatment group. For treatment, TGP (400mg·kg-1) was administered intragastrically every day for 4 weeks. The SS-like symptoms and pathological changes of the SG of mice were compared between the PBS and TGP group. The activation of NLRP3 inflammasome in SG was detected by RT-qPCR, immunohistochemistry and western blot. The SG cells stimulated by lipopolysaccharide (LPS) and adenosine triphosphate (ATP) for activation of NLRP3 inflammasome were treated with or without TGP. Then, NLRP3 inflammasome activation was assessed. The IL-1ß and IL-18 in homogenate of SG, serum and supernatant were detected by ELISA. RESULTS: Compared with balb/c mice, NOD mice showed SS-like symptoms and lymphocyte infiltration in SG, and the expression of NLRP3 inflammasome in SG was significantly increased. The SS-like symptoms were alleviated, and lymphocyte infiltration in SG was reduced, and the level of NLRP3 inflammasome in SG mice was decreased after TGP treatment. TGP also significantly inhibit the activation of NLRP3 inflammasome of SG cells in vitro. CONCLUSIONS: Collectively, our results indicated that TGP alleviates SS through inhibition of the activation of NLRP3 inflammasome of SG. These findings clarified the mechanism underlying the therapeutic effects of TGP on SS, and provided new evidence for the further application of TGP in the treatment of SS.

Radioactive impact of the Fukushima nuclear accident on Shenyang in the northeast of China.

Environmental monitoring was carried out in Shenyang in the northeast of China after the Fukushima Daiichi nuclear accident which was caused by the earthquake and tsunami on 11 March 2011. The fission product radionuclide (131)I was detected as present in the atmosphere on the 20th day after the nuclear accident, while the radionuclides (134)Cs and (137)Cs were found in the atmosphere on the 27th day after the accident. The radionuclides (131)I, (134)Cs and (137)Cs continued to be present in the atmosphere for 25, 4 and 6 days, respectively, with maximum concentrations of 4.60 ± 0.2, 0.29 ± 0.06 and 0.42 ± 0.08 mBq m(-3). The contents of fission radionuclides in vegetables, drinking water and milk from Shenyang were below the detection limits. The atmosphere was slightly contaminated in Shenyang due to the Fukushima nuclear accident, but no contamination was detected in vegetables, milk and drinking water.

Clusters of spatial, temporal, and space-time distribution of hemorrhagic fever with renal syndrome in Liaoning Province, Northeastern China.

BACKGROUND: Hemorrhagic fever with renal syndrome (HFRS) is a rodent-borne disease caused by Hantavirus, with characteristics of fever, hemorrhage, kidney damage, and hypotension. HFRS is recognized as a notifiable public health problem in China, and Liaoning Province is one of the most seriously affected areas with the most cases in China. It is necessary to investigate the spatial, temporal, and space-time distribution of confirmed cases of HFRS in Liaoning Province, China for future research into risk factors. METHODS: A cartogram map was constructed; spatial autocorrelation analysis and spatial, temporal, and space-time cluster analysis were conducted in Liaoning Province, China over the period 1988-2001. RESULTS: When the number of permutation test was set to 999, Moran's I was 0.3854, and was significant at significance level of 0.001. Spatial cluster analysis identified one most likely cluster and four secondary likely clusters. Temporal cluster analysis identified 1998-2001 as the most likely cluster. Space-time cluster analysis identified one most likely cluster and two secondary likely clusters. CONCLUSIONS: Spatial, temporal, and space-time scan statistics may be useful in supervising the occurrence of HFRS in Liaoning Province, China. The result of this study can not only assist health departments to develop a better prevention strategy but also potentially increase the public health intervention's effectiveness.

Investigating the effects of climatic variables and reservoir on the incidence of hemorrhagic fever with renal syndrome in Huludao City, China: a 17-year data analysis based on structure equation model.

BACKGROUND: HFRS is a serious public health problem in China and the study on HFRS is important in China for its large population. The present study aimed to explore the impact of climatic variables and reservoir on the incidence of HFRS in Huludao City, an epidemic focus of the disease in northeastern China. METHODS: Structure Equation Model (SEM), a statistical technique for testing and estimating causal relationships, was conducted based on climatic variables, virus-carrying index among rodents, and incidence of HFRS in the city during the period 1990 to 2006. The linear structural relationships (LISREL) software (Scientific Software International, Lincolnwood, IL) was used to fit SEMs. RESULTS: Temperature, precipitation, relative humidity and virus-carrying index among rodents have shown positive correlations with the monthly incidence of HFRS, while air pressure had a negative correlation with the incidence. The best-fit SEM model fitted well with the data-based correlation matrix, P value was more than 0.56, root mean square error of approximation (RMSEA) equaled to 0, goodness-of-fit index (GFI) was more than 0.99. CONCLUSION: Climate and reservoirs have affected the incidence of HFRS in Huludao City, located in northeastern China. Climate affects HFRS incidence mainly through the effect on reservoir in the study area. HFRS prevention and control should give more consideration to rodent control and climate variations.

Survey of motivation for use of voluntary counseling and testing services for HIV in a high risk area of Shenyang, China.

BACKGROUND: HIV voluntary counseling and testing (VCT) is considered an effective prevention method of HIV infection. In order to understand the VCT environment and enhance the effective delivery of VCT services in a country, an accurate assessment of the current status of VCT services is very important. METHODS: From July 2006 to June 2007, we conducted a cross-sectional survey using a face to face interview among 2676 VCT clients from a high risk area in Shenyang city, China. RESULTS: The major demographic characteristics among 2,676 VCT clients were: 41.1% were in the age range 20 to 30 years; 73.1% were males; and 67.1% had attained the level of junior high school education. The primary information source for VCT services was mass media like television (TV) and newspaper in 88.9%. 34.3% were afraid of the result of infection which was the main barrier to accept VCT services among 540 participants answering the question. 75.2% were motivated by recently acquired knowledge about HIV. 47.9% had 3 or more male sex partners, 62.3% had used condoms sometimes, and 14.5% had been infected with a STD. 2.8% of the participants identified themselves as men who have sex with men (MSM). The main demographic characteristics of MSM did not differ from the total group of participants except with respect to age: 63.5% reported having one male sex partner in the preceding 12 months, 44.6% reported never using condoms in the preceding 12 months, and only 2.7% reported a history of sexually transmitted disease. CONCLUSION: Public education offered by health workers in hospitals, private clinics and other medical institutions needs to be strengthened. Given the results from this study, we recommend: (1) making VCT a routine part of health services, especially in areas where many high-risk individuals live; (2) improving the information sources and increasing the understanding of HIV and HIV-infected individuals; (3) enhancing international collaboration in strategic planning, technical assistance, and protocols to translate policy into effective action; (4) supporting Chinese non-government organizations (NGOs) in playing a significant role in the battle against AIDS.

Teacher Support and Mental Well-Being in Chinese Adolescents: The Mediating Role of Negative Emotions and Resilience.

Teacher support has been shown to enhance adolescent mental health. However, the effects of negative emotions and resilience in the relationship between teacher support and mental well-being in adolescents are still unknown. This study investigated (a) the mediating role of negative emotions in the relationship between teacher support and mental well-being, (b) the mediating role of resilience in the association between teacher support and mental well-being, (c) the serial mediating role of negative emotions and then resilience in the relationship between teacher support and mental well-being, and (d) the parallel mediating role of the five dimensions of resilience and the three factors of negative emotions in the relationship between teacher support and mental well-being. Participants were 1228 Chinese adolescents (age, M = 15.43 years; 53.09% male). Participants filled out questionnaires regarding teacher support, negative emotions, resilience, and mental well-being. After controlling for age and gender, we found that teacher support, negative emotions, and resilience were significantly linked with mental well-being. Moreover, negative emotions and resilience were found to mediate the relationship between teacher support and adolescent mental well-being, accounting for 5.45 and 30.00% of the total effect, respectively. We also found that teacher support enhances mental well-being by decreasing negative emotions and then increasing resilience. This serial mediating effect accounted for 8.48% of the total effect. Finally, the mediating effect of resilience between teacher support and mental well-being was significantly greater than the mediating effects of the other two indirect effects (negative emotions in the relationship between teacher support and mental well-being, negative emotions and then resilience in the relationship between teacher support and mental well-being). And the parallel mediation model showed that teacher support can promote adolescent mental well-being by increasing goal planning, affect control, and help-seeking behavior, and decreasing depression. These findings suggest a process through which negative emotions can decrease resilience and identify the mediating effects of negative emotions (including the three dimensions of negative emotions) and resilience (including the five factors of resilience) in the relationship between teacher support and adolescent mental well-being.

Investigating the effects of climate variations on bacillary dysentery incidence in northeast China using ridge regression and hierarchical cluster analysis.

BACKGROUND: The effects of climate variations on bacillary dysentery incidence have gained more recent concern. However, the multi-collinearity among meteorological factors affects the accuracy of correlation with bacillary dysentery incidence. METHODS: As a remedy, a modified method to combine ridge regression and hierarchical cluster analysis was proposed for investigating the effects of climate variations on bacillary dysentery incidence in northeast China. RESULTS: All weather indicators, temperatures, precipitation, evaporation and relative humidity have shown positive correlation with the monthly incidence of bacillary dysentery, while air pressure had a negative correlation with the incidence. Ridge regression and hierarchical cluster analysis showed that during 1987-1996, relative humidity, temperatures and air pressure affected the transmission of the bacillary dysentery. During this period, all meteorological factors were divided into three categories. Relative humidity and precipitation belonged to one class, temperature indexes and evaporation belonged to another class, and air pressure was the third class. CONCLUSION: Meteorological factors have affected the transmission of bacillary dysentery in northeast China. Bacillary dysentery prevention and control would benefit from by giving more consideration to local climate variations.

Bacillary dysentery and meteorological factors in northeastern China: a historical review based on classification and regression trees.

The relationship between the incidence of bacillary dysentery and meteorological factors was investigated. Data on bacillary dysentery incidence in Shenyang from 1990 to 1996 were obtained from Liaoning Provincial Center for Disease Control and Prevention, and meteorological data such as atmospheric pressure, air temperature, precipitation, evaporation, wind speed, and the amount of solar radiation were obtained from Shenyang Meteorological Bureau. Kendall and Spearman correlations were used to analyze the relationship between bacillary dysentery and meteorological factors. The incidence of bacillary dysentery was treated as a response variable, and meteorological factors were treated as predictable variables. Software R 2.3.1 was used to execute the classification and regression trees (CART). The model improved the accuracy of the fitting results. The residual sum square error of the regression tree model was 53.9, while the residual sum square error of the multivariate linear regression model was 107.2. Among all the meteorological indexes, relative humidity, minimum temperature, and pressure one month prior were statistically influential factors in the multivariate regression tree model. CART may be a useful tool for dealing with heterogeneous data, as it can serve as a decision support tool and is notable for its simplicity and ease.

Analysis of the geographic distribution of HFRS in Liaoning Province between 2000 and 2005.

BACKGROUND: Hemorrhagic fever with renal syndrome (HFRS) is endemic in Liaoning Province, China, and this province was the most serious area affected by HFRS during 2004 to 2005. In this study, we conducted a spatial analysis of HFRS cases with the objective to determine the distribution of HFRS cases and to identify key areas for future public health planning and resource allocation in Liaoning Province. METHODS: The annual average incidence at the county level was calculated using HFRS cases reported between 2000 and 2005 in Liaoning Province. GIS-based spatial analyses were conducted to detect spatial distribution and clustering of HFRS incidence at the county level, and the difference of relative humidity and forestation between the cluster areas and non-cluster areas was analyzed. RESULTS: Spatial distribution of HFRS cases in Liaoning Province from 2000 to 2005 was mapped at the county level to show crude incidence, excess hazard, and spatial smoothed incidence. Spatial cluster analysis suggested 16 and 41 counties were at increased risk for HFRS (p < 0.01) with the maximum spatial cluster sizes at < or = 50% and < or = 30% of the total population, respectively, and the analysis showed relative humidity and forestation in the cluster areas were significantly higher than in other areas. CONCLUSION: Some clustering of HFRS cases in Liaoning Province may be etiologically linked. There was strong evidence some HFRS cases in Liaoning Province formed clusters, but the mechanism underlying it remains unknown. In this study we found the clustering was consistent with the relative humidity and amount of forestation, and showed data indicating there may be some significant relationships.

PC-SPES inhibits cell proliferation by modulating p21, cyclins D, E and B and multiple cell cycle-related genes in prostate cancer cells.

PC-SPES is an herbal mixture, with evidence of clinical efficacy against prostate cancer (CaP), recently attracting tremendous attention. Using immunoblot and cell cycle specific cDNA array analyses, we investigated effects of PC-SPES on LNCaP, a hormone-dependent prostate cancer cell line. PC-SPES inhibited expression of cyclins D and E, inhibited Rb phosphorylation, switching it to a G1-to-S inhibitory state. Moreover, cDNA array analysis showed that PC-SPES caused up-regulation of p21(WAF1/CIP1) and decreased expression of cyclin B, Nedd8, cdc2, skp1, PCNA, MAD2L1, cyclin H, CKS2, E2F, Rbx1, MCM2, MCM5, Mpp2, Cullin-Cul4A, Cks1p9 and McM7, which are involved in cell cycle progression. Taken together, our results provide a mechanistic explanation for antiproliferative and antitumor effects of PC-SPES, suggesting that induction of CDK inhibitors and downregulation of cyclins leads to dephosphorylation of Rb and growth arrest.

Comparison of Two Hybrid Models for Forecasting the Incidence of Hemorrhagic Fever with Renal Syndrome in Jiangsu Province, China.

BACKGROUND: Cases of hemorrhagic fever with renal syndrome (HFRS) are widely distributed in eastern Asia, especially in China, Russia, and Korea. It is proved to be a difficult task to eliminate HFRS completely because of the diverse animal reservoirs and effects of global warming. Reliable forecasting is useful for the prevention and control of HFRS. METHODS: Two hybrid models, one composed of nonlinear autoregressive neural network (NARNN) and autoregressive integrated moving average (ARIMA) the other composed of generalized regression neural network (GRNN) and ARIMA were constructed to predict the incidence of HFRS in the future one year. Performances of the two hybrid models were compared with ARIMA model. RESULTS: The ARIMA, ARIMA-NARNN ARIMA-GRNN model fitted and predicted the seasonal fluctuation well. Among the three models, the mean square error (MSE), mean absolute error (MAE) and mean absolute percentage error (MAPE) of ARIMA-NARNN hybrid model was the lowest both in modeling stage and forecasting stage. As for the ARIMA-GRNN hybrid model, the MSE, MAE and MAPE of modeling performance and the MSE and MAE of forecasting performance were less than the ARIMA model, but the MAPE of forecasting performance did not improve. CONCLUSION: Developing and applying the ARIMA-NARNN hybrid model is an effective method to make us better understand the epidemic characteristics of HFRS and could be helpful to the prevention and control of HFRS.

[Application of nonlinear autoregressive neural network in predicting incidence tendency of hemorrhagic fever with renal syndrome].

OBJECTIVE: To explore the prospect of nonlinear autoregressive neural network in fitting and predicting the incidence tendency of hemorrhagic fever with renal syndrome (HFRS) , in the mainland of China. METHODS: Monthly reported case series of HFRS in China from 2004 to 2013 were used to build both ARIMA and NAR neural network models, in order to predict the monthly incidence of HFRS in China in 2014. Fitness and prediction on the effects of these two models were compared. RESULTS: For the Fitting dataset, MAE, RMSE and MAPE of the ARIMA model were 148.058, 272.077 and 12.678% respectively, while the MAE, RMSE and MAPE of NAR neural network appeared as 119.436, 186.671 and 11.778% respectively. For the Predicting dataset, MAE, RMSE and MAPE of the ARIMA model appeared as 189.088, 221.133 and 21.296%, while the MAE, RMSE and MAPE of the NAR neural network as 119.733, 151.329 and 11.431% respectively. CONCLUSION: The NAR neural network showed better effects in fitting and predicting the incidence tendency of HFRS than using the traditional ARIMA model, in China. NAR neural network seemed to have strong application value in the prevention and control of HFRS.

Effects of herbal preparation Equiguard on hormone-responsive and hormone-refractory prostate carcinoma cells: mechanistic studies.

The Equiguard is a dietary supplement comprised of standardized extracts from nine herbs, respectively, Herba epimedium brevicornum Maxim (stem and leaves), Radix morindae officinalis (root), Fructus rosa laevigatae michx (fruit), Rubus chingii Hu (fruit), Schisandra chinensis (Turz.) Baill (fruit), Ligustrum lucidum Ait (fruit), Cuscuta chinensis Lam (seed), Psoralea corylifolia L. (fruit), and Astragalus membranaceus (Fisch.) Bge (root). This proprietary product, formulated according to Chinese traditional medicinal concepts, is aimed at restoring harmony in the of the kidney, an organ which Chinese medicinal principles consider to be vital for invigorating as well as maintaining balance of the entire urological system. As the prostate is an integral component of the urological system, we performed in vitro studies to test the effects of ethanol extracts of Equiguard to modulate prostate growth and gene expression. These studies used prostate cancer cells mimicking the androgen-dependent (AD) and androgen-independent (AI) states of prostate carcinogenesis. Results show that Equiguard significantly reduced cancer cell growth, induced apoptosis, suppressed expression of the androgen receptor (AR) and lowered intracellular and secreted prostate specific antigen (PSA), and almost completely abolished colony forming abilities of prostate cancer cells. These data support the interpretation that this herbal formulation contains ingredients that collectively may be efficacious in preventing or treating AD and AI prostate carcinoma. The anti-prostatic activities of Equiguard may stem from its complex composition capable of targeting multiple signal transduction/metabolic pathways, to effectively correct, counteract or circumvent the impaired or dysfunctional mechanisms accompanying different stages of prostate carcinogenesis.

Inhibition of proliferation and expression of AR/PSA by herbal supplement Equiguard in LNCaP cells cultured in androgen-proficient FBS and androgen-deficient charcoal-stripped FBS is correlated with increased serine-15 phosphorylation of the tumor suppressor gene p53.

Use of dietary supplements and botanical products is widely accepted by patients diagnosed with prostate cancer (CaP) as a primary or complementary form of treatment for their medical conditions in the U.S. Yet, the majority of these products have not been rigorously studied with regard to scientific mechanism(s). Because many of the available products are mixtures of multiple extracts derived from plants, some of which are not necessarily native to the U.S., we consider mechanistic studies under defined laboratory conditions to be valuable and essential, not only from the standpoint of standardization and possible contamination with the products, but also in providing insights and scientific evidence for the clinical efficacy some of these products purportedly demonstrate. In previous studies from this laboratory, Equiguard, a composite supplement consisting of standardized extracts from nine Chinese herbs, which was originally formulated to correct physiological decline in kidney functions associated with age, was fortuitously found to display anti-CaP properties. Using a panel of CaP cells, we showed that ethanol extracts of Equiguard significantly inhibited cancer cell growth, induced apoptosis, lowered expression of the androgen receptor (AR), decreased intracellular and secreted prostate-specific antigen (PSA) levels and completely abolished the colony forming activities of CaP cells. Since responsiveness to Equiguard was observed in cells mimicking the androgen-dependent (AD) and androgen-independent (AI) states of CaP, our results raise the interesting possibility that this herbal supplement may potentially prevent, delay or circumvent the onset of AI, and thereby induce chronic instead of terminal CaP. Since androgen ablation therapy (chemical or surgical castration) is the mainstay for localized CaP, we questioned whether Equiguard might still exert the aforementioned activities in experimental settings modeled after androgen ablation. Accordingly, we studied the effects of Equiguard in LNCaP cells, cultured in androgen-proficient (FBS) or -deficient (CS-FBS) media that simulate the hormonal status pre- and post-castration in vivo. Extracts of Equiguard were effective in reducing colony formation, proliferation and PCNA expression of cells cultured in CS-FBS. Moreover, within a concentration range of Equiguard, the prostate-specific genes, PSA and AR, were affected to a similar extent in cells cultured either in FBS or CS-FBS, and were correlated with increased phosphorylation at serine-15 of the tumor suppressor gene p53. These results are consistent with the interpretation that the anti-proliferative and gene modulatory properties of Equiguard are largely independent of the status of androgens in the culture media.

Seroprevalence of antibodies to highly pathogenic avian influenza A (H5N1) virus among close contacts exposed to H5N1 cases, China, 2005-2008.

To assess the extent of highly pathogenic avian influenza (HPAI) A (H5N1) virus transmission, we conducted sero-epidemiologic studies among close contacts exposed to H5N1 cases in mainland China during 2005-2008. Blood specimens were collected from 87 household members and 332 social contacts of 23 H5N1 index cases for HPAI H5N1 serological testing by modified horse red-blood-cell hemagglutinin inhibition and microneutralization assays. All participants were interviewed with a standardized questionnaire to collect information about the use of personal protective equipment, illness symptoms, exposure to an H5N1 case during the infectious period, and poultry exposures. Two (2.3%) household contacts tested positive for HPAI H5N1 virus antibody, and all social contacts tested negative. Both seropositive cases had prolonged, unprotected, close contact with a different H5N1 index case, including days of bed-care or sleeping together during the index case's infectious period, and did not develop any illness. None of the 419 close contacts used appropriate personal protective equipment including 17% who reported providing bedside care or having physical contact with an H5N1 case for at least 12 hours. Our findings suggest that HPAI H5N1 viruses that circulated among poultry in mainland China from 2005-2008 were not easily transmitted to close contacts of H5N1 cases.

[Genetic characterization analysis on epidemic rubella virus strains isolated in Liaoning from 2007 to 2012].

To analyze the genetic characterization of epidemic rubella virus strains isolated in Liaoning from 2007-2012, a total of 145 rubella virus strains were isolated using Vero/Slam cell line from the patients' throat swabs during rubella outbreaks and sporadics cases in Liaoning Province from 2007 to 2012. Fragments of 945 nucleotides containing 1E gene from 145 rubella virus isolates were amplified by RT-PCR, the PCR products were sequenced and analyzed. Based on the 739 nucleotides of 1E gene, the phylogenetic trees were constructed with 32 WHO rubella reference strains of 13 genotypes downloaded from GenBank and 145 rubella virus strains. The results showed that the 145 rubella virus strains in 2007 -2012 belonged to genotype 1E, nucleotide acids and amino acids similarities were 97.2%-100.0% and 97.6%-100.0%, respectively. Compared to the 1E reference strains(Rvi/ Dezhou.CHN/02, RVi/MYS/01), the nucleotide acids and amino acids similarities were 96.6%-99.2% and 98.2%-100.0%, respectively except for one amino acid change (Val246-Ala246) of RVi/Shenyang. Liaoning. CHN/13.11/13, and Asp262-Asn262 of RVi/Shenyang. Liaoning. CHN/13.11/4 and RVi/Liaoyang. Liaoning. CHN/26. 11/2. there had no change found in the important antigenic epitope sites, the hemagglutination inhibition and neutralization epitopes of the other rubella viruses. All the 145 strains isolated had the same amino acid change (Leu338--Phe338) in E1 protein. These findings suggested that genotype 1E of rubella virus was the predominant genotype in Liaoning province. the rubella prevailed in recent six years was mainly caused by rubella viruses genotype 1E with multi-transmission routes.

[SH protein genetic characterization analysis of wild-type mumps virus isolated in Liaoning province from 2008 to 2011].

A total of 13 mumps virus strains were isolated using Vero/Slam cell line from the patients' throat swabs and urines during mumps outbreaks and sporadics in liaoning Province from 2008 to 2011. Fragments of 316 nucleotides containing SH genes from 13 mumps virus isolates were amplified by RT-PCR, the PCR products were sequenced and analyzed. Based on the 316 nucleotides of SH gene, the phylogenetic trees were constructed with WHO mumps reference strains downloaded from GenBank and 13 mumps virus strains. It showed that the 12 mumps virus strains in 2008-2011 belonged to F genotype, nucleotide acids and amino acids similarities were 94.9%-100.0% and 83.3%-100.0%. Compared to the F reference strains, nucleotide acids and amino acids similarities were 92.4%-97.2% and 96.5% 84.2%, indicating that liaoning mumps strains had large genotype variation. Furthermore, compared to other genotype mumps strains, 6 nucleotides mutations (C(Nt65), C(Nt105), G(Nt137), C(Nt192), C(Nt239), G(NT262)) in SH gene were shared in all of liaoning F genotype mumps viruses. However these mutations were not found in other genotypes of mumps viruses. Conserved amino acids in SH protein of some liaoning mumps strains had changed (S to P at position 2 P to L at position 6 T to N at positon 23 L to P/R at position 48). The genotype specific triplet IML (at position 28-30) was changed to TMP in strain 2008-001-007 2011-015 s, nucleotide acids and amino acids similarities of strain 2008-001-07 were 87.5% and 79.8%, compared to F reference strains respectively, and 96.8% and 97.4% respectively compared to the G reference strains, in dicating strain 2008-001-07 belonged to G genotype,which was first found in mainland China.

[Isolation of the Culex flavivirus from mosquitoes in Liaoning Province, China].

A flavivirus, Culex flavivirus, was first isolated from Chinese mosquitoes with high sequences similarities to those of flaviviruses found in America and Japan. In this study, a total of 48 pools of field-collected mosquitoes were sampled from Dandong of Liaoning Province, China during July to September of 2011. Six isolated viruses showing cytopathic effect (CPE) in C6/C36 cells were tested by reverse transcription polymerase chain reaction(RT-PCR)using Flavivirus genus--specific primers and Culex flavivirus-specific primers and the positive PCR-product was sequenced and compared with the sequences of 10 isolates from GenBank. Phylogenetic tree of NS5 and enevelop genes of flavivirus were constructed. The GenBank accession numbers of NS5 gene were JQ409188, JQ409186, JQ409187, JQ409191, JQ409189 and JQ409190. The GenBank accession numbers of envelope gene were JQ065883, JQ065882, JQ065881, JQ065879,JQ065877 and JQ065878.

[Genetic characterization of wild-type mumps virus isolated in Liaoning Province].

Three mumps virus strains were isolated using Vero/Slam cell line from the patients' throat swabs and urines during mumps outbreaks and sporadic period in Liaoning province from 2008. Fragments of 1028 nucleotides including SH genes from 3 mumps virus isolates were amplified by RT-PCR, the PCR products were ligated into pMD19-T vector and cloned to JM109 cell. By blue-white selection, the positive white clones were sequenced and analyzed. Based on the 316 nucleotides of SH gene, the phylogenetic analyses were processed with WHO mumps reference strains downloaded from GenBank and 3 mumps viruses strains. It wan shown that the 3 mumps virus strains isolated in 2008 belonged to F genotype, 3 strains (LN-2008-001-06, LN-2008-001-07 and LN-2008-001-10) showed a nucleotide and amino acid similarity of 98.7%-100% and 94.7%-100% respectively. Two strains (LN-2008-001-06 and LN-2008-001-10) had the same sequence completely. Comparing to the F reference strains, the 3 mumps virus strains' nucleotide and amino acid similarity of 92.4%-96.2% and 84.2%-94.7% respectively. Due to the limited strain numbers, whether the F genotype was the predominant circulating genotype can not be determined. The surveillance on the mumps virus in Liaoning should be therefore strengthened.