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Objective To explore the clinical significance of the changes of platelet membrane glycoprotein levels in peripheral blood of patients with gastric cancer and its influencing factors. Methods The levels of platelet membrane glycoproteins (CD62p and CD63) in peripheral blood of 87 patients with gastric cancer were measured by flow cytometry (FCM). 40 healthy people were served as control. Statistical analysis was performed by using the SPSS 10.0 software package. Results The levels of platelet membrane glycoproteins in peripheral blood in patients with gastric cancer [CD62p: (4.15 ± 1.42) % and CD63 (4.72 ± 1.54) %] were significantly higher than controls [CD62p (1.54 ± 0.44) % and CD63 (1.23 ± 0.34) %] P < 0.01. The levels of platelets membrane glycoproteins in peripheral blood in patients with gastric cancer were significantly associated with clinical stage [CD62p (2.80 ±0.63) versus (4.76 ± 1.25); CD63 (3.28 ± 0.70) versus (5.36 ±1.37], operation [(CD62p:3.49 ± 1.21) versus (4.86 ± 1.28), (3.98 ± 1.31) versus (5.51 ± 1.38)], metastasis [CD62p (4.94 ±1.07) versus (3.31 ± 1.25); CD63 (5.55 ± 1.19) versus (3.82 ± 1.38)], while were not influenced by gender and pathological types. Conclusion Detecting the levels of platelets membrane glycoprotein in peripheral blood may dynamically monitor the progression and prognosis of gastric cancer. [ABSTRACT FROM AUTHOR]

Objective To compare the proportion and biological characteristics of side population cells and non side population cells before and after chemoradiotherapy. Methods We selected 60 cases diagnosed as cervical cancer and staged IB2-IIIB, 30 cases were given paclitaxel at a dose of 175 mg/m² combined with cisplatin at a dose of 80 mg/m², the chemotherapy was repeated every 21 days. The human cervical cancer cells were obtained from fresh human cervical cancer tissue before chemotherapy and after two chemotherapy processes respectively. Another 30 cases received radiotherapy, the human cervical cancer cells were obtained from fresh human cervical cancer tissue before radiotherapy and two weeks after radiotherapy (wide field radiation DT20Gy/10F) respectively. Flow cytometry was used to isolate and compare the proportion and biological characteristics between side population cells and non side population cells from fresh human cervical cancer tissue before and after chemoradiotherapy. Results The proportion of SP cells had statistically significant difference between before and after treatment (before and after chemotherapy 1.63 ± 0.50% vs 7.96 ± 1.39%, before and after radiotherapy 1.63 ± 0.46% vs 4.51 ± 0.72%) .The proportions of ABCG2, CD 133, CD43 and P63 of SP cells had no statistically significant difference between before and after treatment, but the proportions of Ki-67 and MDR of SP cells were significantly higher than pretherapy and statistically significant difference (Ki-67 before and after chemotherapy 14.77 ± 1.97% vs 23.49 ± 2.23%, before and after radiotherapy 14.61 ± 1.40 vs 23.10 ± 1.82%; MDR before and after chemotherapy 59.31 ± 11.13% vs 68.91 ± 9.94%, before and after radiotherapy 57.42 ± 11.22% vs 59.37 ± 10.73%) . There were no statistically significant differences in cell cycle or cell apoptosis of SP cells. The proportions of Ki-67 and MDR of non SP cells were significantly higher than pre-therapy and had statistically significant difference ( Ki-67 before and after chemotherapy 40.48 ± 3.35% vs 46.29 ± 3.71%, before and after radiotherapy 40.22 ± 4.70% vs 45.82 ± 4.55%; MDR before and after chemotherapy 17.76 ± 1.61% vs 34.36 ± 3.15%, before and after radiotherapy 17.66 ± 1.71% vs 19.21 ± 1.98%) . The proportions of non SP cells after therapy in G0/ G1 and S were significantly lower, but in G2/M and apoptosis were significantly higher. Conclusions Chemoradiotherapy might kill mainly non SP cells, the SP cells after chemoradiotherapy still keep the characteristics of cancer stem cells, and are resistant to chemoradiotherapy. This study lay the basis for further targeted therapy of cervical cancer stem cell. [ABSTRACT FROM AUTHOR]

We undertook this study in order to explore the effect of reduced glutathione (GSH) on the physicochemical indexes and non-volatile and volatile metabolites of Chardonnay wine during a five-month storage period after the end of alcoholic fermentation. Five different concentrations of GSH were added into the wine after the end of alcoholic fermentation. The changes of physicochemical indexes, total phenolic content and color during the storage period were measured. Widely targeted metabolomic analysis and volatile metabolomic analysis based on ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) and gas chromatography-mass spectrometry (GC-MS) were performed on wine samples with and without the addition of 20 mg/L GSH. The results showed that during the five-month storage period, the addition of different concentrations of GSH had no significant effect on the volatile acid content, total acid content and pH of white wine (P > 0.05) but significantly influenced its total phenol content, total flavonoid content, color and free SO2 content. The total phenol content of the wine sample added with 30 mg/L GSH was highest after storage for five months, and the color index of the wine sample added with 20 mg/L GSH was lowest. The addition of GSH could delay the decrease of free SO2 content. A total of 693 non-volatile metabolites and 816 volatile metabolites were identified. Based on variable importance in projection (VIP) greater than 1.0 and fold change (FC) equal to or greater than 1.4, a total of 33 non-volatile differential metabolites and 22 volatile differential metabolites were selected. Among the 32 non-volatile metabolites, 17 were down-regulated and the rest 16 were up-regulated. Among the 22 volatile metabolites, three were down-regulated and the other 19 were up-regulated. Some phenolic acids, fatty acids and amino acids were associated with changes in volatile metabolites.

For selective separation of 90Sr from HLLW, dodecyl benzenesulfonic acid (DBS) was applied to modified the 4′,4′(5″)-di(tert-butylcyclohexano)-18-crown-6 (DtBuCH18C6) and a novel silica-based crown ether was synthesized. The successful synthesis of adsorbent was estimated by characterization methods such as SEM, TG-DTA and FT-IR. The leakage of DtBuCH18C6 from the adsorbent is less than 2.03% in 7 mol/L HNO3, suggesting a good chemical stability of adsorbent. DBS worked as a counter ion to enhance the complex formation between DtBuCH18C6 and Sr(Ⅱ), enhancing the Kd value at 0.5 mol/L HNO3 to 10 times (355.02 cm3/g), and a synergistic effect with nitrate ions was found at higher HNO3 concentration. The selective adsorption of Sr(Ⅱ) is proved great, which follows the pseudo-second-order kinetic model and Langmuir model, and the maximum adsorption capacity reaches 42.13 mg/g. Using 1 mol/L HNO3 solution as eluent, 79% of the adsorbed Sr(Ⅱ) can be eluted from the adsorbent (ratio of eluent vs adsorbent is 0.9 L/g, contact time is 5 h), resulting in effective Sr(Ⅱ) recycle and reuse of adsorbent.

Peritoneal adhesion is one of the common complications after abdominal operation, which could seriously affect the quality of life in patients. Although the development of modern surgical technology and the improvement of doctors' operation level have reduced the incidence of peritoneal adhesion to a certain extent, due to the lack of special treatment drugs, the therapeutic effect still cannot meet the expectations and requirements of clinicians and patients. Traditional Chinese medicines(TCM) have unique advantages and remarkable curative effect in the treatment of peritoneal adhesion, and they can play an important role in regulating multiple pathological links. However, the relevant researches and product development of TCM against peritoneal adhesion have not attracted enough attention from industry scholars. As for the related work that has been carried out, most of the studies on the efficacy and mechanism are not thorough and systematic enough, seriously restricting the industrial development in this field. In this paper, the efficacy and mechanism were systematically described and summarized based on the review of papers in the recent years, so as to provide a reference for the thorough study of TCM in the prevention and treatment of peritoneal adhesions, and promote the deep development and industrialization process of related products.

Aim: To induce the expression of structure protein VP1 and its C terminus of foot-and-mouth disease virus (FMDV) serotype SAT2 in E.coli and analyze their reactivities with FMDV positive antiserum., Methods: The plasmid pGEM-SAT2P1 carrying the VP1 coding region of FMDV serotype SAT2 isolated from South African was used as template for RT-PCR to get the coding fragment of VP1 and its C terminus. The fragments were then cloned into expression vector pET-30a(+) to get recombinant plasmids pET-SAT2VP1 and pET-SAT2VP1C. The recombinant plasmids were transformed into E.coli BL21(DE3)pLysS and induced by IPTG to express VP1 and its C terminus protein. The expressed VP1 and its C terminus were then purified by Ni-NTA His Bind Resin affinity chromatography and analyzed by Western blot. New Zealand rabbits were immunized to prepare polyclonal antibodies against VP1 and VP1C. The antisera were obtained and polyclonal antibody was characterized by ELISA., Results: SDS-PAGE demonstrated that VP1and its C terminus expressed in the E.coli transformants had a molecular weight of 33000 and 19000 and contained in the inclusion body. Purified VP1 and its C terminus was obtained by Ni-NTA His Bind Resin affinity chromatography and a single clear band appeared in the SDS-PAGE gel. Western blot analysis showed that the purified VP1 and VP1 C terminus could react with bovine antiserum against the same serotype FMDV without cross-reactivity with the negative bovine serum., Conclusion: Rabbit polyclonal antibodies against VP1 and VP1C were successfully prepared, the titers of which were above 1:12800 and had obvious specificity.

A study of the continuous precipitation in Guiyang has been taken during October 30 and November 7. The results showed that the range of pH was between 3.65 and 7.20, and volume-weighted mean was 4.24. SO4(2-) was the main anion and volume-weighted mean was 119.06 microeq x L(-1), accounting for 80.63% of total anions. While Ca2+ was the main cation and volume-weighted mean was 48.87 microeq x L(-1), accounting for 48.16% of total cation and NH4+ was 38.38 microeq x L(-1), 37.82%. Good correlations between Mg2+, Ca2+, NH4+ and SO4(2-) are observed, which may exist as MgSO4, CaSO4 and (NH4)2SO4 in the rainwater in Guiyang City. Different ions of the atmosphere had different scavenged ways and rate in the precipitation process. In the initial stage of precipitation, the ions such as Mg2+, which were rich in coarse particles, were quickly removed from atmosphere. However the ions such as NH4+ and SO4(2-), which were rich in fine particles, could remain in the atmosphere for a longer time.

Aim: To clone and express the ligand binding domain (LBD) cDNA of porcine integrin beta3 as foot-and-mouth disease virus (FMDV) receptor and prepare its polyclonal antibody., Methods: The LBD cDNA of porcine beta3 was obtained from the lung tissue of pig infected with FMDV by RT-PCR, and the recombinant plasmid pGEM/beta3LBD was constructed. After digested with BamH I/Xho I, the beta3LBD fragment was subcloned into prokaryotic expression vector pGEX 4T-1. The recombinant expression plasmid pGEX/beta3LBD was constructed and transformed into E.coli BL21(DE3). The recombinant porcine beta3LBD protein was expressed after IPTG induction and purified from total protein of BL21(DE3). The rabbits from New Zealand were immunized with the purified fusion protein to prepare polyclonal antibody, which was identified by Western blot and ELISA., Results: The 507 bp cDNA of porcine beta3LBD encoded a polypeptide of 169 amino acids. The similarity of nucleotide sequence beta3LBD between pigs and cattle, human being, chimpanzees, rhesus monkeys, horses, dogs, Norway rats, mice, chickens was 90.3%, 92.3%, 92.1%, 91.3%, 90.5%, 90.3%, 87.8%, 85.2%, 79.5%, respectively. The beta3LBD gene of mammals exhibited high sequence homology. The recombinant beta3LBD protein was expressed efficiently as inclusion body after IPTG induction and was approximately 44000. The titer of the polyclonal antibody against the purified beta3LBD protein was about 1:12 800 by ELISA., Conclusion: The gene cloning and expression of beta3LBD and the preparation of its polyclonal antibody lay a foundation for further research into the interaction of FMDV with beta3 subunit of porcine integrin.

Aim: To prepare the monoclonal antibodies against VP1 protein of type AsiaI foot-and mouth disease virus (FMDV) and identify the characterization., Methods: Three cell of hybridization that strains secreted monoclonal antibody(mAb) against type AsiaI FMDV were produced by fusing mouse myeloma cells (Sp2/0) with spleen cells from BALB/c immunized with the purified recombinant VP1 protein., Results: The three hybridized cell lines reacted with cattle type AsiaI FMDV, the titres of ascetic fluids of the three mAbs ranged from 1:10(5) to 1:10(6) indirect ELISA showed. Among the three mAbs,1B8 belonged to IgG1, 5E1 and 5E2 pertained to IgG2a. Besides, the three strain antibodies stably excreted antibodies and reacted with type AsiaI FMDV., Conclusion: VP1 protein of FMDV could replace FMDV can be use to replace FMDV to prepare mAbs.

Receptors play a crucial role in determining the pathogenesis and tissue tropism of virus. Foot-and-mouth disease virus (FMDV) has been showed to use four integrins, alphavbeta1, alphavbeta3, alphavbeta6 and alphavbeta8 as receptors to initiate infection. In this study, the porcine integrin alphav gene was cloned by RT-PCR from the lung tissue of healed pig infected experimently with FMDV, and compared its nucleotide and deduced amino acid sequence with the av gene of other animals. The 3141bp cDNA of bovine integrin alphav encodes a polypeptide of 1046 amino acids consisting of a 30-residue putative signal peptide, a 955-residue ectodomain, a 29-residue transmembrane domain, and a 32-residue cytoplasmic domain. The ectodomain contains 11 potential N-linked glycosylation sites (NXT/NXS), 2 calcium binding domains (DX[D/N] XDGXXD) and 18 cysteine residues. The nucleotide sequence similarities of integrin alphav between pig and cattle, human, rheses monkey, house mouse, chicken, dog are 93.3%, 91.5%, 91.4%, 85.6%, 73.2% and 89.9% respectively; and the amino acid sequence similarities are 96.3%, 94.6%, 94.1%, 90.8%, 81.6% and 93.8%, respectively. The alphav gene of cattle and pig exhibited the highest sequence homology. It is possible that host tropism of FMDV may related to divergence in receptors among different species.

The full-length genomic sequence of foot-and-mouth disease virus (FMDV) Asia1/YNBS/58 strain was determined by RT-PCR and compared with other 17 reference strains. The results showed that the complete genome of Asia1/YNBS/58 was 8164nt long including a 1061-nt 5' untranslated region (UTR), a 6990-nt open reading frame (ORF), and a 113-nt 3'UTR. The homology analysis indicated that the UTR regions and non-structural proteins were more conserved than the structural proteins in FMDV. VP1 exhibited the lowest conservation and VP4 was exceptionally conserved. The VP1-, VP2-, and VP3-based phylogenetic trees were divided into distinct clusters according to different serotypes, while the other gene-based phylogenetic trees exhibited some degree of intercross among serotypes. This study is the first description of the full-length genomic sequence of FMDV Chinese serotype Asia1.

In order to study the roles of integrin beta6 in Foot-and-Mouth Disease Virus infection, pig integrin beta6 was firstly molecularly cloned from RNA of the tongue and lung of recovered pig infected experimentally with foot-and-mouth-disease virus (FMDV), and was compared with the beta6 gene of other animals available in GenBank at nucleotide and amino acid leves. GeneBank association number of the beta6 gene is EF432729. Pig integrin beta6 gene (2367bp) encodes a polypeptide of 788 amino acids consisting of 9 potential N-linked glycosylation sites, 3 Glycosaminoglycan attachment sites, a cGMP-dependent protein kinase phosphorylation site, 10 Protein kinase C phosphorylation sites, 2 EGF-like domains and 2 cysteine-rich regions. Pig integrin beta6 subunit has a 26-residue putative signal peptide, a 681-residue ectodomain, a 29-residue transmembrane domain, and a 52-residue cytoplasmic domain. 11 mutant nucleotides were found in beta6 gene coding region and 9 amino acids were changed. The nucleotide sequence similarity of integrin beta6 gene between rheses monkey, mouse, Norway rat, dog, guinea pig, human, bovine, sheep is 79.5%, 84.9%, 85.4%, 85.2%, 88.7%, 90.1%, 91.9% and 91.9%, and the amino acid sequence similarity is 93.5%, 88.2%, 88.5%, 88.3%, 91.0%, 92.8%, 93.3% and 93.4% respectively. This study will lay a foundation for understanding the interactions of FMDV with receptors.

The complete gene encoding the structural protein of FMDV(VP1) was subcloned into expression vector pPROex-HT, resulting in the fusion expression plasmid pPROexHT-VP1. After transformed into E. coli BL21(DE3) and induced by IPTG, the fusion protein was expressed in high level. Western blot was performed to confirm that the expressed fusion protein could specifically react with antiserum against FMDV. Based on the fusion protein further purified, a novel indirect ELISA (VP1-ELISA) was developed to detect FMDV antibody in pigs. Comparison between VPl-ELISA and the government standard kit (liquid phase block ELISA) showed the two methods had 96.25 percent agreement by detecting 80 serum samples, indicating that the indirect VP1-ELISA was specific and sensitive.