Two fragments G1 and G2 of the glycoprotein G gene of bovine respiratory syncytial virus(BRSV) were selected for expression in Escherichia coli based on the analysis of glycoprotein G by DNA Star software.Then the two fragments of glycoprotein G were amplified by PCR with synthesized G gene of BRSV as the template.The amplified fragments G1 and G2 are 570bp and 308bp in length,respectively.The PCR products were cloned into pET30a vector and expressed in soluble form in E.coli after induction of cultured E.coli with IPTG.Both of the recombinant proteins G1 and G2 were purified by immobilized Ni ion affinity chromatography under native conditions.Then the purified proteins were analysed by Western blotting.The results showed that the purified recombinant protein G1 retained good antigenicity and specificity.But the purified recombinant protein G2 didn't possess biological activity.Antibodies against BRSV were detected in suspected bovine serum samples in China by using indirect ELISA and Western blotting with the purified recombinant protein G1.The purified recombinant protein G1 might be used as antigen for establishing serological methods for diagnosis of BRSV infection.And the purified recombinant protein G1 might also be used for preparing polyclonal and monoclonal antibodies for research on biological functions of glycoprotein G of BRSV.

Leaves of Chrysanthemum morifolium were potential medicinal resource. The present study aims to estimate the main bioactive components: total flavonoids (TF), galuteolin (GA), quercitrin (QU), chlorogenic acid (CA) and 3 ,5-O-caffeoylquinic acid ( CQ), which were considered to be the main effective components, in leaves of C. morfolium cultivars in China. The TF content was estimated hy UV-VIS spectrophotometry, while GA, QU, CA, and CQ were quantitatively determined by HPLC. The highest TF content (7. 13% w/w) was found in cultivar Wan Cong (Shexian county). Cultivar Da Bo ( Bozhou county) had the highest GA content (33. 45 mg - g-1); Cultivar Hong Xin (Sheyang county) contained the highest QU content (29.25 mg · g(-1)); Cultivar Chang Ban (Sheyang county) had the highest CA content (13.14 mg ·(-1)). The maximum CQ content (7.35 mg · g(-1)) was observed in culti- r Da Yang ( Tongxiang county). Different cultivars of C. morfolium had significant difference in components, but the leaf and capitulum of C. morifolium. were found to possess similar chemical compositions. The high content of bioactive components in several cultivars suggested the potential utilization of C. morifolium leaves.
China ; Chromatography, High Pressure Liquid ; Chrysanthemum ; chemistry ; growth & development ; Drugs, Chinese Herbal ; analysis ; Plant Leaves ; chemistry

OBJECTIVETo determine vacA dominant genotypes of Helicobacter pylori in patients with peptic ulcer (PU) or chronic gastritis (CG).

METHODSH.pylori strains were isolated from mucosa samples of gastric antrum and corpus of patients suffering from PU (n=29) or CG (n=34), 126 strains of H.pylori were selected for PCR to detect s and m regions in vacA gene of the isolates. Parts of the amplification products were sequenced after T A cloning. The correlation between infection or coinfection with different vacA genotypes of H.pylori and different gastroduodenal diseases was further analyzed.

RESULTSThe positive amplification products of vacA gene, s and m regions, were found in the DNA samples of all the isolates. In these products, s1a/m1, s1a/m2, s1a/m1b and s1a/m1b m2 genotypes of vacA gene were detected and s1b and s2m1a genotypes absent. Proportions of the s1a/m1, s1a/m2, s1a/m1b and s1a/m1b-m2 genotypes were 7.1% (9/126), 61.9% (78/126), 29.4% (37/126) and 1.6% (2/126), respectively. 17.5% (11/63) of the patients were confirmed to be coinfected with different genotype H.pylori strains. No statistical differences were found in the distribution of different genotype H.pylori strain infection in the gastric diseases (P>0.05). In comparison with the reported sequences of H.pylori strain 60190 with s1a genotype and strain 87-203 with m2 genotype, homologies of the nucleotide sequences of s1a PCR products from 6 strains of H.pylori isolates and m2 PCR products from 4 strains of H.pylori isolates were 93.15% approximate, equals 94.86%and 93.63% approximate, equals 97.61%, respectively.

CONCLUSIONH.pylori with s1a/m2 or s1a/m1b are the dominant genotypes in the PU or GC patients in Zhejiang area. The nucleotide sequences of partial amplification products from the vacA dominant genotypes of H.pylori show high homology compared with the reported sequences. Part of the patients may be coinfected with different vacA genotypes of H.pylori.

Adolescent ; Adult ; Aged ; Bacterial Proteins ; genetics ; Base Sequence ; Child ; Female ; Genotype ; Helicobacter pylori ; genetics ; Humans ; Male ; Middle Aged

OBJECTIVETo screen and identify proteins that interact with hepatitis C virus NS3 by means of T7-phage display system.

METHODSHepatitis C virus NS3 was expressed by prokaryotic expression and used as a selected molecule to biopan the T7 select human liver cDNA library; the selected positive clones were identified using DNA sequencing and analyzed with BLAST program in GenBank.

RESULTSAfter BLAST analysis in all the positive clones, the proteins which interacted with the hepatitis C virus NS3 were found to be serpin peptidase inhibitor, clade A, member 1 (SERPINA1) and cyclophilin-LC.

CONCLUSIONT7-phage display system is a convenient, rapid and effective method for screening interacting proteins. The proteins thus selected will provide an important means for studying the pathogenesis and carcinogenesis of HCV.

Cell Line ; Gene Library ; Hepacivirus ; metabolism ; Humans ; Peptide Library ; Protein Interaction Mapping ; methods ; Viral Fusion Proteins ; genetics ; isolation & purification ; metabolism ; Viral Nonstructural Proteins ; genetics ; metabolism

The study is aimed to ensure the quality and safety of medicinal plants by using ITS2 DNA barcode technology to identify Corydalis boweri, Meconopsis horridula and their close related species. The DNA of 13 herb samples including C. boweri and M. horridula from Lhasa of Tibet was extracted, ITS PCR were amplified and sequenced. Both assembled and web downloaded 71 ITS2 sequences were removed of 5. 8S and 28S. Multiple sequence alignment was completed and the intraspecific and interspecific genetic distances were calculated by MEGA 5.0, while the neighbor-joining phylogenetic trees were constructed. We also predicted the ITS2 secondary structure of C. boweri, M. horridula and their close related species. The results showed that ITS2 as DNA barcode was able to identify C. boweri, M. horridula as well as well as their close related species effectively. The established based on ITS2 barcode method provides the regular and safe detection technology for identification of C. boweri, M. horridula and their close related species, adulterants and counterfeits, in order to ensure their quality control, safe medication, reasonable development and utilization.
Base Sequence ; China ; Corydalis ; chemistry ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; chemistry ; genetics ; DNA, Ribosomal Spacer ; chemistry ; genetics ; Molecular Sequence Data ; Nucleic Acid Conformation ; Papaveraceae ; chemistry ; classification ; genetics ; Phylogeny ; Plants, Medicinal ; chemistry ; classification ; genetics

OBJECTIVETo determine cagA/vacA dominant genotypes of Helicobacter pylori in patients suffering from chronic gastritis (CG) or peptic ulcer (PU), and to understand the correlation of different genotype H. pylori infection, coinfection and the gastroduodenal diseases.

METHODSH. pylori strains were isolated from antrum and corpus samples on 42 patients with CG and 36 patients with PU. Polymerase chain reaction was used to detect cagA and the s and m regions of vacA in 156 H. pylori isolates from both antrum and corpus. The distribution of H. pylori genotypes and coinfection in CG and PU was analyzed.

RESULTSAlmost all of the isolated H. pylori strains were cagA positive. In region of vacA, only one genotype of signal region (s1a) and four genotypes of the middle region (m1, m2, m1b and m1b-m2) were found. The proportions of s1a/m1, s1a/m2, s1a/m1b, s1a/m1b-m2 and coinfection of multiple H. pylori strains in 78 isolates from antrum samples were 6.4%, 55.1%, 26.9%, 1.3% and 3.8%; and the related proportions of those from corpus samples were 6.4%, 53.8%, 25.6%, 3.8% and 5.1%, respectively. Sixteen (20.5%) patients had multiple H. pylori strains with different cagA and vacA genotypes, and multiple samples were better than single sample taken from one stomach to increase the positive proportion of coinfection.

CONCLUSIONcagA(+) s1a/m2 was the dominant genotype of H. pylori in the CG or PU patients followed by cagA(+) s1a/m1b in the Zhejiang area of China. Some of the patients were coinfected with multiple H. pylori strains of different cagA and vacA genotypes. However, there was no significant correlation between the genotypes or mixed infection with multiple strains, CG or PU.

Adolescent ; Adult ; Aged ; Antigens, Bacterial ; genetics ; Bacterial Proteins ; genetics ; Child ; China ; Female ; Genes, Dominant ; Genotype ; Helicobacter Infections ; microbiology ; Helicobacter pylori ; classification ; genetics ; isolation & purification ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction

OBJECTIVETo investigate the expressions of PTEN and Caspase-3 proteins in human breast carcinoma, and to evaluate their clinicopathological implications during the tumorigenesis and progression of breast cancer.

METHODSThe expressions of PTEN and Caspase-3 proteins in 95 cases of breast cancer and 15 cases of benign breast diseases were investigated immunohistochemically. Correlations between the expression of PTEN protein, Caspase-3 protein, and clinicopathological features of breast cancers were analyzed.

RESULTSThe loss expression rate of PTEN protein in tumor tissues was significantly higher than that in benign breast diseases (33.7% vs. 0, P < 0.01). Analysis of the clinicopathological features showed that PTEN expression level was negatively correlated with TNM stage, histological grade, axillary lymph node status, recurrence, and metastasis (P < 0.05). The positive expression level of Caspase-3 was negatively correlated with TNM stage (P < 0.01), but not related with histological grade, axillary lymph node status, recurrence, or metastasis (P > 0.05). In addition, the expression of PTEN protein had significantly positive correlation with the expression of Caspase-3 protein in breast cancer (P < 0.01).

CONCLUSIONThe combination detection of PTEN and Caspase-3 may serve as an important index to estimate the pathobiological behavior and prognosis of breast cancer.

Adult ; Aged ; Animals ; Breast Neoplasms ; metabolism ; pathology ; Caspase 3 ; metabolism ; Female ; Humans ; Kaplan-Meier Estimate ; Middle Aged ; PTEN Phosphohydrolase ; metabolism ; Young Adult

Current studies have demonstrated that SLC38A1 proteins play a causal role in neoplastic cell transformation.The twofold aim of this study was to provide insight into whether a variance in the expression of SLC38A1 exists between human colorectal cancer and healthy human tissues and to determine how silencing or overexpressing the SLC38A1 gene could affect the proliferation,viability and migration of colorectal cancer cells.Immunohistochemical staining was used to analyze the expression of SLC38A1 in colorectal cancer tissues and adjacent normal mucosa in 77 patients who underwent surgical resection.The expression of SLC38A1 in colorectal cancer tissues and cell lines was detected using RT-PCR and Western blotting.Two colorectal cancer cell lines SW480 and HCT116 were used to examine whether silencing SLC38A1 with siRNA and overexpressing SLC38A1 with shRNA could affect cell viability and migration.As a result,the SLC38A1 protein was very low or undetectable in the normal colon mucosa.In contrast,strong staining of SLC38A1 protein was found in the cytoplasm in 79.2％ colorectal cancer samples.More pronounced SLC38A1 expression in colorectal cancer tissues was significantly associated with tumor node metastasis (TNM) stage.Inhibition of SLC38A1 reduced tumour growth and suppressed proliferation and migration of SW480 cells.In contrast,overexpression of SLC38A1 had the opposite effects on HCT116 cells.S LC38A1 is overexpressed in colorectal cancer,which suggests that it is associated with tumour progression.These results encourage the exploration of SLC38A1 as a target for intervention in colorectal cancer.

On the basis of the chromogenic reaction between Hg and CuI, a semi-quantitative solid sampling Hg analyzer comprising the catalytic furance, Hg testing tube, air pump and smart cellphone was developed. White carrier 101 was chosen as the adsorbent for CuI to react with Hg from the catalytic furnace. The established Hg analyzer can not only visually recognize the coloration when Hg exceeding the limit standard, but also semi-quantitatively detect the Hg content in cosmetics fast using a smart cellphone and RGB analysis software, after direct solid sampling introduction of cosmetics sample. The instrumental detection limit ( LOD) of mercury was 50 ng, the linearity ranged from 50 ng to 2500 ng, the linear regression coefficient ( R2) was higher than 0. 97, and the RSD of the corresponding RGB values was 6% ( n=11 ) . Nine real cosmetics samples were measured by the established method, whose relative differences of Hg contents with that by the standard method (Safety Technical Specification for Cosmetics, 2015 edition) were less than 10% . The whole analytical time can be controlled within 5 min. The established instrumental method is simple, fast, accurate and visual, and extremely suitable to fast and on-site monitoring of Hg in cosmetics samples.

AIM To evaluate saponins in Paris Polyphylla Smith var.chinensis (Franch.) Hara from twentyone growing areas.METHODS HPLC was adopted in the content determination of pennogenins { pennogenin-3-O-β-D-glu (1→3) [α-L-rha (1→2)]-β-D-glu (PGGR),polyphyllinsⅥ,Ⅶ,H} and dioscins (gracillin,polyphyllins Ⅰ,Ⅱ,Ⅴ),then SPSS 22.0 software was used for principal component analysis and cluster analysis.RESULTS Pennogenin was the main kind of Paris Polyphylla var.chinensis saponins,and PGGR,polyphyllins Ⅰ,Ⅴ,Ⅶ,H were the main effective constituents of this medicinal material.The comprehension scores of seven growing areas were more than zero,which was in sequence of Bazhong ＞ Qingchuan ＞ Yanjin ＞ Tongjiang ＞ Nanchuan ＞ Chongzhou ＞ Hongya.Twenty-one batches of samples were divided into four types,which was similar to cluster analysis results.CONCLUSION Bazhong,Qingchuan,Chongzhou,Hongya,Tongjiang in Sichuan,Yanjin in Yunnan,and Nanchuan in Chongqing are suitable for cultivating Paris Polyphylla var.chinensis,which can guarantee the stability of saponins.