Purpose The aim was to examine c-MET,ALK,ROS1 variants in advanced non-small cell lung cancer (NSCLC) patients,and to analysis the association of c-MET,ALK,ROS1 variants with the clinical and pathological features.Methods The c-MET,ALK,ROS1 were detected by fluorescence in situ hybridization (FISH) in the 91 cases of NSCLC specimens.The correlation of c-MET gene amplification with clinicopathological features and the ALK,ROS1 fusions was analyzed.Results The positive rate of c-MET gene amplification was 8.79％ (8/91),the positive rates on male and female were 1.82％ and 19.4％,respectively.In ＜ 60-years-old and ≥60-years-old NSCLC patients,the positive rates were 7.5％ and 8.89％,resepectively.The positive rate was higher in stage Ⅲ than stage Ⅳ (9.62％ vs 7.69％),the c-MET gene amplification was detected in 9.2％ adenocarcinoma patients but none in squamous carcinoma patients.The detection rates of ALK fusions and ROS1 fusions were 10％ and 13.3％,respectively.One patient was detected the coexistence of MET with ROS1 fusion.Conclusion The c-MET gene amplification is correlated with gender,but not with age,histological types and clinical stages.C-MET amplification,ALK fusions and ROS1 fusions are almost no coexistence,but not completely mutually exclusive.To they knowledge,this is the first case report the coexistence of MET amplification with ROS1 fusion in NSCLC.

BACKGROUNDThe vaccination of mice with DNA encoding single candidate antigens has failed to induce significant protection against Schistosoma japonicum (S. japonicum) challenge infections. In this study, we evaluated the feasibility of using a multivalent DNA vaccine which co-expressed S. japonicum integral membrane protein Sj23 and murine cytokine IL-12 to induce protective immune responses.

METHODSThe plasmid pVIVO2-IL12-Sj23, a eukaryotic expression vector expressing Sj23 and murine IL-12 simultaneously, was constructed, identified, and tested for expression in vitro. Its ability to protect against S. japonicum challenge infections was analyzed according to worm reduction rate and egg reduction rate after vaccination of BALB/c mice. The serum levels of specific IgG antibody were determined by enzyme-linked-immuno sorbent assay (ELISA) and Western blot analysis. Using cultured spleen cells, IFN-gamma and IL-4 post-stimulation were quantified by ELISA. The phenotypes of splenocyte populations were analyzed by flow cytometry (FCM).

RESULTSThe plasmid DNA pVIVO2-IL12-Sj23 was proven to express well in vitro by transient transfection of HEK-293 cells. Immunization resulted in a worm reduction rate of 45.53% and egg reduction rate of 58.35%. ELISA and Western blot analysis indicated that immunized mice generated specific IgG against Sj23. Spleen cells showed significant increases in IFN-gamma but decreases in IL-4. No significant differences in CD4+ and CD8+ subgroup ratios were observed after the challenges.

CONCLUSIONSThe multivalent DNA vaccine pVIVO2-IL12-Sj23 is sufficient to elicit moderate but highly significant levels of protective immunity against challenge infections. Cytokine IL-12, as a gene adjuvant, was able to enhance the Th1 responses and, hence, the protective immunity.

Animals ; Antibodies, Helminth ; blood ; Antigens, Helminth ; genetics ; immunology ; CD4-CD8 Ratio ; Cytokines ; biosynthesis ; Helminth Proteins ; genetics ; immunology ; Interleukin-12 ; genetics ; immunology ; Male ; Membrane Proteins ; genetics ; immunology ; Mice ; Mice, Inbred BALB C ; Schistosoma japonicum ; immunology ; Th1 Cells ; immunology ; Vaccination ; Vaccines, DNA ; immunology

BACKGROUNDRecently congenital infection with Schistosoma japonicum (S. japonicum) has been demonstrated in pigs, rabbits, mice and dogs. We explored the rabbit as an animal model for the congenital infection of schistosomiasis japonica and assessed the effect of a congenital S. japonicum infection on the resistance of rabbit kittens to a postnatal challenge infection.

METHODSSixteen pregnant New Zealand white rabbits were infected with a single dose of S. japonicum cercariae. The exposed animals were divided into three groups according to the gestation age at the time of infection. Diagnosis of prenatally acquired S. japonicum infection in the rabbit kittens was primarily based on serological tests in combination with parasitological and histopathological findings. Congenitally infected kittens were challenged percutaneously with 100 S. japonicum cercariae to assess the effect of a congenital S. japonicum infection on kitten resistance to a postnatal challenge infection.

RESULTSThe overall prevalence of congenital infection in offspring of infected mothers was 20% (12/60). The congenital infection rate in group L (late gestation) was much higher than in group E (early gestation) and group M (mid-gestation) (P <0.05). After a postnatal challenge infection, prenatally infected kittens had a 54.66% worm reduction rate, 41.45% egg reduction rate, and 51.76% granuloma size reduction rate compared to naïve kittens.

CONCLUSIONSThis study demonstrates the possibility of congenital infection of S. japonicum in rabbits and the resistance of congenitally infected kittens to a postnatal challenge infection. These results have important implications not only for epidemiological investigations, but also in designing government control programs for schistosomiasis.

Animals ; Antibodies, Helminth ; blood ; Female ; Immunoglobulin M ; blood ; Infectious Disease Transmission, Vertical ; Male ; Pregnancy ; Rabbits ; Schistosomiasis japonica ; congenital ; immunology ; parasitology

In order to analyze the temporal distribution characteristics of human rabies in China and explore the incidence forecasting,R3.3.2 software were used to establish the optimal ARIMA model of monthly incidences from 2004 to 2015 in Chi-na.Then the monthly incidences from January to November 2016 were forecasted with the model and the prediction accuracy was evaluated.The annual incidences of human rabies in China were decreasing tendency since 2007,and the monthly incidence reached highly during August to October.The optimal model was ARIMA(2,1,1)(2,0,0)12and the mean absolute error (MASE)was 0.755.The mean relative error was 15.61% when predicting the incidences from January to November 2016.In conclusion,there exists seasonal variation for human rabies in China and ARIMA model can be applied for the short-term fore-casting.

Objective Toinvestigatethestatusoftick-borneRickettsiaeinfectionsamongmurine-likeanimalsin differentareasofZhejiangprovince.Methods Liverandspleensamplesofmurine-likeanimalscapturedthroughnight trapping method were collected from Anji,Jinhua and Tiantai County according to their geographic locations and historical detection of Rickettsiae .Nest-PCR tests were used to determine the presence of the 16S rRNA genes of Anaplasma and Ehrlichia ,and the heat shock protein genes (groEL)of Rickettsiae (including typhus and spotted fever group)and Orientiainthesesamples.Results Atotalof851murine-likeanimalsbelongingto14specieswerecaptured.The predominant species were Rattus confucianus (30.32%),Apodemus agrarius (18.80%) and Thallomys paedulcus (1 1.75%)and they were significantly different among three areas (P<0.05 ).48 Rickettsia positive were found in 562 tested samples with the positive rate of 8.54%,among which the percentage of Anaplasma,typhus group Rickettsia, Orientia,Ehrlichia and spotted fever group Rickettsia were 3.38%,1.78%,1.78%,1.07% and 0.53% respectively. The positive rates of Anaplasma in Jindong (4.76%)and Anji (4.27%)were significantly higher than that in Tiantai (P<0.05 )while the spotted fever group Rickettsia were found only in Tiantai County.Moreover,Rattus confucianus-the predominant species of Zhejiang Province-had the highest infection rate of tick-borne Rickettsiae up to 14.97%.Co-infections with several Rickettsiae were existed among the same species.Conclusion Rickettsiae infections exist widely among different areas of Zhejiang province and the positive rates are significantly different among species.

Objective ToinvestigateanimportedcaseofZikavirusdiseaseinWenzhouCity,toexplorethesourceof infection,to analyse the problems during surveillance and diagnosis,and to provide evidence for prevention and control of Zikavirusdisease.Methods Byfieldinvestigation,medicalrecordsofthepatientwerecollectedandaface-to-face interview was conducted;blood,urine and saliva samples within 48 h were collected for RNA detection;assess the risk of diseasespreadingbymosquitosurveillanceandtakecontrolmeasures.Results Theonsetofthediseasewasonthepatient's way home from aboard on February 21st,2016.Before then,She lived in Paramaribo,the capital of Suriname in South America,which is an epidemic area of Zika virus.The patient had exposure history,with red,diffuse rash on her head,face, neck and limbs,and once was diagnosed as allergic eruption because of no fever.The two blood samples were both tested negative yet We detected urine and saliva samples showed positive.With the evidence above,the patient was finally diagnosed as an imported Zika virus case.The spreading risk of this case was low due to the low temperature.Conclusion there may be cases without fever or with blood sample tested RNA negative.As the weather gets warmer,missed diagnosis should be aware of,or the risk of spreading through mosquitos would increase.Therefore,in order to further improvement should lie in mosquito control and disease surveillance.

OBJECTIVETo analyze the geographical distribution and risk factors of HIV-1 subtypes in Yunnan province.

METHODSBlood samples from 1319 HIV positives were collected in Yunnan Province from 2001 to 2006. The nested polymerase chain reaction was used to amplify the gag (p24)-protease fragments from RNA extracted from plasma or sera. The sequences were used for subtype determination by phylogenetic tree analysis.

RESULTSAmong 1319 samples studied, the subtypes has been successfully obtained from 644 samples that were constituted of seven subtypes: CRF08_BC, CRF07_BC, CRF07/08_BC, CRF01_AE, C, B' and URFB/C. C/CRF07_BC/CRF08_BC were distributed in the whole province, but CRF01_AE were mainly distributed in the boarding areas with Myanmar such as Dehong, Baoshan, Xishuangbanna and Puer. Moreover, injecting drugs users accounted for 61.6% (270/438) among C/CRF07_BC/CRF08_BC infections, while only 8.5% (15/177) among CRF01_AE infections.

CONCLUSIONOur data indicated that at least seven subtypes were identified in Yunnan province, the relationship between subtypes and transmission routes were analyzed, and the geographic difference of subtypes was also observed.

China ; DNA, Viral ; Genotype ; HIV Infections ; transmission ; virology ; HIV-1 ; classification ; isolation & purification ; Humans ; Sequence Analysis, DNA

Background/Aims: Esophagogastric junction adenocarci-noma (EJA) is a malignant tumor associated with high mor-bidity and has attracted increasing attention due to a rising incidence and low survival rate. Pathological biopsy is the gold standard for diagnosis, but noninvasive and effective tests are lacking, resulting in diagnoses at advanced stages.This study explored the diagnostic value of insulin-like growth factor binding protein 7 (IGFBP7) in EJA. Methods: A total of 120 EJA patients and 88 normal controls were recruited, and their serum levels of IGFBP7 were measured by enzymelinked immunosorbent assay. Receiver operating character-istic (ROC) curve analysis was used to assess the diagnostic value, and Pearson chi-square analysis was used to evaluate the correlation between IGFBP7 and clinical parameters. Ka-plan-Meier survival analysis was carried out to assess the ef-fect of IGFBP7 on overall survival (OS). Results: The levels of IGFBP7 were higher in both early- and late-stage EJA patients than in normal controls (p<0.001). The area under the ROC curve for EJA patients was 0.794 (95% confidence interval [CI], 0.733 to 0.854), with a cutoff value of 2.716 ng/mL, a sensitivity of 63.3% (95% CI, 54.0% to 71.8%) and a specific-ity of 90.9% (95% CI, 82.4% to 95.7%). For the diagnosis of early-stage EJA, the same cutoff value and specificity were obtained, but the sensitivity of IGFBP7 was 54.3% (95% CI, 36.9% to 70.8%). Patients with low IGFBP7 protein expres-sion had lower OS than those with high expression (p=0.034).The multivariate analysis showed that IGFBP7 is an inde-pendent prognostic factor for EJA (p=0.011). Conclusions Serum IGFBP7 acts as a potential diagnostic and prognostic marker for EJA.

Objective:This studu aims to investigate the effect of aqueous extract of modified Xiao Xianxiongtang on the epithelial mesenchymal transition（EMT） and the change of its invasion and migration ability of human gastric cancer MGC-803 cells mediated by transforming growth factor-β₁（TGF-β₁），and to explore the mechanism of regulating Wnt5a/Ca²⁺/ activated T-cell nuclear factor（NFAT） signaling pathway to inhibit EMT and invasion and metastasis of MGC-803 cells. Method:TGF-β₁（10 μg·L^-1）was used to induce EMT and the invasion and metastasis model of human gastric cancer MGC-803 cells. Transwell chamber experiment， scratchhealing experiment， Western blot and immunofluorescence assay were used to detect cell invasion and migration ability， expression of EMT marker protein and key protein expression of Wnt5a/Ca²⁺/NFAT signaling pathway， and intracellular Ca²⁺ concentration. Result:Compared with the blank group， TGF-β₁ could significantly enhance the invasion and migration ability of MGC-803 cells（P<0.01）， down-regulate the level of E-cadherin（P<0.01）， up-regulate protein expressions of N-cadherin， Snail and Vimentin（P<0.01）， and induce cell Wnt5a， calcineurin （CaN）， total protein of activated T-cell nuclear factor 1（NFAT1）， up-regulation of phosphorylated proteins p-NFAT1 and NFAT1 nucleoprotein and intracellular accumulation of Ca²⁺（P<0.01）. Compared with the TGF-β₁ group， modified Xiao Xianxiongtang （10， 20， 40 mg·L^-1） could significantly inhibit this phenomenon，and 40 mg·L^-1 had the best effect（P<0.05，P<0.01）.The specific inhibitors of Wnt5a/Ca²⁺/NFAT signaling pathway （R）-（+）-Bay-K-8644 and modified Xiao Xianxiongtang （40 mg·L^-1） could significantly inhibit theinvasion and migration of MGC-803 cells mediated by TGF-β₁， up-regulate the level of E-cadherin， and down-regulate expressions of N-cadherin， Snail， Vimentin， Wnt5a， CaN and NFAT1 proteins and reduce the intracellular accumulation of Ca²⁺（P<0.05，P<0.01）.Moreover， （R）-（+）-Bay-K-8644 combined with modified Xiao Xianxiongtang （40 mg·L^-1） had stronger inhibitory effect（P<0.05，P<0.01）. Conclusion:These results suggest that modified Xiao Xianxiongtang can inhibit the EMT mediated by TGF-β₁ via Wnt5a/Ca²⁺/NFAT signaling pathway，thereby reducing the invasion and migration ability of MGC-803 cells.

In order to comprehensively monitor the dynamic change of Paeonia lactiflora planting area,the investigation of P. lactiflora planting area in Dangshan was carried out. It can provide reference for the planting detection of P. lactiflora in Huaibei Plain.Based on remote sensing technology,this paper extracts the planting area of P. lactiflora in Dangshan in 2018 by using the minimum distance method,maximum likelihood method,parallel hexahedron method and Mahalanobis distance method,using the remote sensing image of ZY-3 Satellite as the data source,and makes a comparative analysis with the results. The results show that the maximum likelihood method is better than the other three methods. This method can provide reference for remote sensing monitoring of P. lactiflora planting area in China.
China ; Paeonia ; Remote Sensing Technology