The sequence encoding an E2 main antigen glycoprotein of the C strain of classical swine fever virus (CSFV) was highly expressed in the host cell E. coli BL21-CodonPlus (DE3)-RIL using the pGEX-4T-1 expression vector and the soluble recombinant product was purified with Glutathione Sepharose TM<'4B> by centrifugation. The soluble recombinant protein showed good immune reactions and was confirmed by Western blot using anti-CSFV-specific antibodies. Then an indirect ELISA with the purified E2 protein as the coating antigen was established to detect antibody against CSFV. The result revealed that the optimal concentration of coated antigen was 0.6 μg/well and the optimal dilution of serum was 1:80. The positive cut-off value of this ELISA assay was OD<,tested serum>/OD<,negative serum>≥2.1- The E2-ELISA method was evaluated by comparison with the indirect hemagglutination test (IHAT). When a total of 100 field serum samples were tested the sensitivity and specificity were 90.3% and 94.7% respectively. Specificity analysis showed that there were no cross-reactions between BVD serum and the purified E2 protein in the E2-ELISA.

Objective To investigate the clinical characteristics between elderly and non-elderly patients with ulcerative colitis (UC) in order to provide the base for diagnosis and management of the disease.Methods Two hundred and four patients with UC admitted to Shengjing Hospital from January 2009 to December 2011 were enrolled in this study.The patients were divided into elderly group (≥60 years of age) and non-elderly group(＜ 60 years of age).The clinical manifestation,laboratory examination indices and endoscopic feature were recorded and analyzed.Results Among the 204 cases with UC,33 (16.2％,33/204) were elderly patients,and 171 (83.8％,171/204) were non-elderly patients.Onset peak of U C was ranged from 30-39 years old.There was no significant difference in terms of other indices of clinical presentations,extraintestinal manifestations,complications,clinical and endoscopic score between two groups.The rate of abdominal distension,constipation,difficult defecation,alteration of intestinal flora,hypokalemia and hypoproteinemia in elderly group were 36.4％,9.1％,6.1％,66.7％,45.5％,36.4％ respectively,higher than that in in nonelderly group (14.6％,1.2％,0.6％,48.0％,14.0％,20.5％ respectively),and there was significant differences between groups (x2 =8.808,7.260,5.724,3.876,17.660 and 3.942 respectively,P ＜ 0.05).Endoscopic examination revealed that the colonic lesion parts in elderly patients was limited in left-side colon and protosigmoid,which was different from that of no-elderly group (P =0.044).Conclusion The elderly patients showed a similar clinical manifestation with non-elderly patients.However,the colonic lesion parts in elder group was limited to left-side colon and protosigmoid.They also had the higher rate of hypoproteinemia,hypokalemia and alteration of intestinal flora,which needed to pay more attention.

Objective To investigate the effects of 15-deoxy-?12,14-prostaglandin J2(15d-PGJ2) and DDP on the growth of human pulmonary carcinoma PLA-801D cells and the mechanisms of apoptosis.Methods The human pulmonary carcinoma PLA-801D cells were selected and added to each well of 96-well place and cultivated for 24 h.Then the cells were treated with different concentrations of 15d-PGJ2(0,5,10,20,40 and 80 ?g?L-1) or 15d-PGJ2 combined with DDP(3 mg?L) for 24 h.0 ?g?L-1 15d-PGJ2 group was control group.The morphological changes of cells were observed under inverted microscope.Microculture tetrazolium(MTT)dye was applied to detect the proliferation of the human pulmonary carcinoma PLA-801D cells treated with 15d-PGJ2 and DDP.Diphenylamine assay(DPA) was used to evaluate the activation.Flow cytometry assay(FCM) was used to detect the apoptosis proportion and the changes of cell cycle.Results When the human pulmonary carcinoma PLA-801D cells were treated with low-concentration 15d-PGJ2 alone(5,10 and 20 ?g?L-1),no significant difference was observed in the inhibitory rate of cell growth and the apoptotic indexes such as the apoptosis proportion,the percent of DNA fragmentation and the activity of caspase-3 compared with control group(P

Objective: Combining with national drug sampling program,to evaluate the current quality situation and problems of compound Yinqiao and paracetamol and chlorphenamine maleate capsules by testing and analyzing 78 batches of samples collected from the realm of drug production and circulation all over the country in 2015.Methods: As the current standard could not control the product quality, and combined the prescribed examination with exploratory research, the method of HPLC was used to determine the contents.GC and TLC was respectively used to identify peppermint oil and study on forsythin.Near infrared spectrum database was established by near-infrared spectroscopy (NIRS), which provided the basis for rapid testing.The results of the prescribed examination and exploratory research were statistically analyzed.Results: There were significant differences in the results of the prescribed examination and exploratory research.Conclusion: The results of exploratory research show that there are many defects in the statutory standards.The product quality of different manufactures was divers, and it is necessary to guide them to improve the preparation process and the quality.More exclusive, accurate and sensitive methods should be used to comprehensively control the quality.

Objective:To investigate the predictive value of gadobenate dimeglumine (GD-BOPTA) multi-phase enhanced MRI for the expression of cytokeratin19 (CK19) in hepatocellular carcinoma (HCC).Methods:A total of 153 patients of HCC confirmed by pathology from June 2016 to February 2020 in First Affiliated Hospital of Fujian Medical University were enrolled retrospectively. According to the post-operative pathology, the patients were divided into CK19-negative group ( n=122) and CK19-positive group ( n=31). All the patients underwent MRI scan and Gd-BOPTA multi-phase enhanced scan before operation. MRI features on Gd-BOPTA MRI were compared between two groups. The qualitative indicators included tumor morphology, mosaic signs, intratumoral hemorrhage, intratumoral fat, non-rim arterial phase hyper-enhancement (APHE), non-peripheral washout, targetoid manifestation, enhanced capsule, corona enhancement, DWI signal, vascular invasion and hepatobiliary phase (HBP) enhancement. The quantitative indicator of tumor-to-liver signal ratio (SR) on HBP was recorded. The χ 2 test or Fisher exact probability method was used to compare the qualitative parameters between two groups, and student′s t test or Mann -Whitney U test was used for quantitative data. Predictive parameters were identified by univariate and multivariate logistic regression analysis to predict the value of the expression of CK19. The ROC curve was used to analyze the diagnostic efficacy of MRI parameters. Results:There were statistically significant differences between CK19-positive and CK19-negative groups ( P<0.05) in alpha fetoprotein, tumor morphology, non-rim APHE, non-peripheral washout, targetoid manifestation, corona enhancement, HBP enhancement and SR. Multivariate logistic regression analysis showed tumor morphology, corona enhancement, HBP enhancement and SR were independent predictors of CK19 expression in HCC. The area under the ROC curve of the combined four indicators for predicting CK19 expression in HCC was 0.823, and the sensitivity and specificity were 80.7% and 75.4%, respectively. Conclusions:Gd-BOPTA multi-phase enhanced MRI has an important value in the prediction of the expression of CK19 in HCC. The combination of signs of HBP can improve the prediction efficiency of CK19.

Objective To observe the clinical effect of lithium carbonate on hyperthyroidism.Methods Forty-seven cases of hyperthyroidism with abnormal hepatic function and/or leucopenia were recruited from January 2010 to January 2012.Thyroid function,adverse reactions,and clinical outcome of patients treated with lithium carbonate were observed and recorded before and after treatment.Results Thyroid function,liver function,and leucopenia in all patients were improved markedly after treatment.Among the treated patients,9 cases resulted in complete remission and had the drug discontinued,11 cases received radioactive iodine therapy afterwards,6 cases underwent surgical treatment,16 cases continued to use the same drug,and 5 cases were lost during follow-up.No obvious adverse reaction appeared during the treatment.Conclusions Lithium carbonate can be effectively used in patients with hyperthyroidism complicated by leukopenia or liver damage.It is also indicated during preparation for radioactive iodine or surgical treatment in patients with thyrotoxicosis.

This study was aimed to identify and distinguish Metacordyceps liangshanensis recorded by the Sichuan Province Traditional Chinese Medicine Standard from its adulterants and its relative species by combining ITS and COI barcode sequences in order to study the feasibility of this new method. After extracting DNA of 28 species of Cordyceps samples, DNA were amplified and sequenced. And then, ITS and COI sequences were received. Codon-Code Aligner V3.7.1 and Mega 5.0 were used to analyze the variable site and construct the N-J tree. The results showed that the minimum ITS inter-specific K-2P distance was relatively higher than the maximum intra-specific K-2P distance. The inter-specific sequence divergence between M. liangshanensis and its adulterants exhibited high while intra-specific sequence divergence exhibited low. And COI one was the same case. N-J tree of both ITS and COI indicated that same genus belonged together and each species belonged to relatively independent branch. It was concluded that based on the ITS and COI gene, the technology of DNA barcode can be an excellent identification of M. liangshanensis, its adulterants and its relative species. It provided technical support for the further research on species molecular identification and phylogenetics of Cordyceps .

Using in vitro everted gut to investigate the intestinal absorption of the extracts from Polygonum orientale at different concentration. UPLC-MS/MS was used to detect the content of protocatechuic acid, isoorientin, orientin, vitexin, cynaroside, quercitrin, kaempferol-rhamnoside in different intestinal segments, then compared the results with the absorption of chemical components of extractive P. orientale in each intestinal segments, and calculated the absorption parameter. We took the statistic analysis with SPSS statistic software. The influence significance of each factors were analyzed to describe the character of absorption. The absorption of each component is linearity in different intestinal segments and different dose, and the square of coeficient correlation exceed 0.95, which consistent with zero order rate process. The K(a) increase along with the raised dosage of the extractive P. orientale (R2 > 0.95), indicated it is the passive absorption; different intestinal segments have different absorption. And the absorption trend in intestinal is duodenum, jejunum, ileum are greater than the colon. As ingredients are selectively absorbed in intestinal sac, the everted intestinal sac method is selected to assess the intestinal absorption charcteristics of ingredients of extractive P. orientale.
Animals ; Drugs, Chinese Herbal ; pharmacokinetics ; Gastrointestinal Tract ; metabolism ; Intestinal Absorption ; Male ; Polygonum ; chemistry ; metabolism ; Rats ; Rats, Sprague-Dawley

In this study,a synthesized quadruple antigenic epitope gene region of the classical swine fever virus (CSFV)E2 glycoprotein was expressed in E.coli to a obtain target protein.This target protein was used as a coating antigen to establish an indirect ELISA for specifically detecting anti-CSFV antibodies in serum samples from pigs.The P/N cut-off value of this assay was 1.92 by receiver operating characteristic curve(ROC)analysis based on 30 negative sera and 80 positive samples.The test gave 97.5% sensitivity and 96.7% specificity compared with the indirect hemagglutination(IHA)test.The inter-assay and intra-assay coefficients of variation (CVs)for 16 sera were both ≤6.8%.No cross-reactivity between the coating antigen and anti-bovine viral diarrhoea virus(BVDV)antibodies was observed.

Purpose To study the malignant transformation after treating rat oval cell line ( WB-F344 ) with chemical carcinogen N-methyl-N′-nitro-N-nitrosoguanidine ( MNNG) . Methods WB-F344 cells were cultured with MNNG for severe times. The biological characteristics of induced cells were detected through the following methods:to check proliferation activity by flow cytometry analysis, to examine malignant transformation degree of induced cells by soft agar assay and tumor formation in NOD/SCID mice, and to investi-gate the transcriptional and protein levels of hepatocellular carcinoma marker GGT, GST-P by real time-PCR. Results Oval cells in-duced by MNNG showed changes in biological characteristics and malignant molecular markers. Conclusion Hepatic oval cells model is successfully established, which can be confirmed by tumor formation in NOD/SCID mice.