1.Detection of Epstein-Barr Virus(EBV) and expression of EBV lytic genes in EBV-positive gastric carcinomas
Yun WANG ; Bing LUO ; Xiaofeng WANG
China Oncology 2001;0(03):-
Purpose:To explore the relationship between Epstein-Barr virus(EBV) infection and gastric carcinoma and the role of EBV lytic genes in the tumorigenesis of EBV-gastric carcinomas. Methods:185 gastric carcinoma tissues and 185 corresponding para-carcinoma tissues were tested for EBV genome by polymerase chain reaction (PCR)-southern analysis. EBV-encoded small RNA 1(EBER1) of the PCR positive specimens was detected by in situ hybridization (ISH). Gastric carcinoma with positive EBER1 signals was confirmed as EBV-positive gastric carcinoma. RT-PCR and Southern blotting were used to detect the expression of EBV lytic genes (immediately early genes BZLF1 and BRLF1, early genes BARF1 and BHRF1, late genes BcLF1 and BLLF1) in EBV-positive gastric carcinomas. Results:There were 13 EBV positive samples in gastric carcinomas (7.03%). No EBV positive sample was found from corresponding para-carcinomas. The difference of the EBV positivity was significant between carcinoma and corresponding adjacent carcinoma tissues(? 2= 11.0769,P=0.0009). In our series, age, pathological differentiation, clinical stages, lymph node metastasis and location of cancer were not different between EBV-positive and EBV-negative gastric carcinomas in (P=0.973, 0.141, 0.259, 0.586, 0.062, respectively), while sex was significantly different between EBV-positive and EBV-negative gastric carcinomas(? 2=5.2317,P=0.021). The EBV positivity of male was higher than that of female. Of the 13 EBV-associated samples, 7 exhibited BcLF1 transcript and 2 exhibited BHRF1 transcript. The transcripts of BZLF1 were detected in 6 cases, and those of BARF1 also in 6 cases. No BLLF1 and BRLF mRNA were detected in the 13 EBV-positive samples. Conclusions:EBV infection is associated with the development of gastric carcinoma. Lytic EBV infection occurs in part of the EBV-associated gastric carcinomas, and early genes BARF1 and BHRF1 may play an important role in the tumorigenesis of the EBV-positive gastric carcinoma.
2.Detection of antibodies to Epstein-Barr virus with ELISA based on fusion capsid protein
Qingfang QIU ; Yun WANG ; Bing LUO
Chinese Journal of Laboratory Medicine 2009;32(9):1002-1005
and 98.6% (436/442), respectively. Conclusion The ELISA based on fusion viral capsid proteins is sensitive, specific and accurate method for determining antibodies to VCA of EBV for both clinical diagnosis and epidemiology studies.
3.Study on the antiviral effects of Stichopus japonicus Selenka glycosaminoglycans (SJ-GAG) on HSV-Ⅰ
Bing LUO ; Zhongbing MA ; Xiaofeng WANG ; Yun WANG
Chinese Journal of Marine Drugs 2000;0(06):-
Objective To study the antiviral effects of SJ-GAG on herpes simplex virus Ⅰ(HSV-Ⅰ) in vitro as well as its protective effect on infected mice.Methods The Vero cells were exposed to HSV-Ⅰ SM44 and different concentrations of SJ-GAG,respectively.Cytopathic effect(CPE) was observed and qunatitative PCR was used to evaluate the antiviral effect of SJ-GAG.In vivo experiment,the mice were infected with HSV1 intracranial vaccination and followed SJ-GAG intragastric administration 4h later to test the protection of SJ-GAG on HSV1 infected mice.Results When the concentration of SJ-GAG was above 1.6mg?mL-1,it showed cytotoxicity.When the concentration was among 0.25~0.2mg?mL-1,it expressed marked antiviral effect without cytotoxicity.SJ-GAG could prolong the survival duration of infected mice and decrease the mortality rate significantly.The protection of SJ-GAG showed a dose-effect relationship.Conclusion SJ-GAG has antiviral effect and shows some protective effect on HSV-1 infected mice.
4.Roles of Th_1/Th_2 type cytokines in experimental autoimmune neuritis
Yun WU ; Hua-Bing WANG ; Wei-Zhi WANG ;
Chinese Journal of Neurology 2005;0(09):-
Objective To establish the model of P2 peptide-induced experimental autoimmune neuritis(EAN)in rats and explore the roles of Th_1/Th_2 type eytokines in EAN.Methods Lewis rats were grouped into EAN rats and control rats.The EAN rats were immunized by injection into both hind footpads of inoculums containing 100 ?g or 200 ?g of P2_(57-81)peptide and FCA while the control rats were immunized with FCA only.Clinical scores were compared at the maximum of disease.Supernatant productions of IFN- ?, IL-4 and IL-10 secreted by lymphocytes and obtained on day 14 after the immunization were examined. Histopathological assessment of sciatic nerves was made.Results Peak clinical scores of P2_(57-81)200 ?g (3.6?0.3)group were significantly higher than P2_(57-81)100 ?g group(2.2?0.6,P
6.Influence of Liang-Xue-Huo-Xue (LXHX) Capsule on Apoptosis of Cultured Keratinocytes
Li-Yun SUN ; Bing-Xu DENG ; He WANG ; Kai CHEN
Chinese Journal of Dermatology 2003;0(10):-
Objective To investigate the pharmacological mechanism of LXHX capsule. Methods The skin specimens from psoriasis patients were examined with TUNEL (terminal deoxynucleotidyl trans-ferase-mediated dUTP-biotin nick end labeling) technique to detect the apoptosis of keratinocytes. PCNA (proliferating cell nuclear antigen) detecting kits was used, too. Then, the flow cytometry, with AnnexinV/PI and PI dyeing method, was used to analysis the effect of LXHX capsule on apoptosis in cultured keratinocytes. Results There was an increasing of both cell apoptosis and proliferation in psoriatic epidermis and LXHX capsule could induce apoptosis. Conclusions The keratinocyte apoptosis and proliferation both increased in psoriasis, which reach a new balance in a higher level. LXHX capsule could induce apoptosis in vitro, which may be one of the pharmacological mechanisms of LXHX in treating psoriasis.
7.Expression of recombined human endothelial monocyte-activating polypeptideⅡand determination of its activity
Yun GAO ; Liang-Hua WANG ; Na REN ; Ming-Juan SUN ; Ai-Yun GUO ; Bing-Hua JIAO ;
Chinese Journal of Cancer Biotherapy 1995;0(03):-
Objective: To chine and express the recombinant human endothelial monocyte-activating polypeptide-Ⅱ(EMAP-Ⅱ)and identify its anti-tumor biological activities.Methods: EMAP-Ⅱ_(147-312)was expressed by the expression vector pMAL-p2x and E.coli BL-21 and the product was purified.The production of tissue factor(TF)in human umbili- cal vein endothelial cell ECV-304 mediated by the recombinant EMAP-Ⅱwas determined by chemiluminescence sub- strate.The promoting effect of recombinant EMAP-Ⅱon TNF?-induced ECV-304 cell.Apoptosis was determined by flow cytometry.Its inhibitory effect on human pancreaic cancer cell SW1990 proliferation was determined by MTT method. Results:DNA sequencing verified that EMAP-Ⅱwas correctly cloned.The molecular mass of the protein identified by SDS-PAGE was consistent with the theoretic value.The productivity of recombinant EMAP-Ⅱwas 500?g per 1 g bacteria (wet mass).The purified product induced expression of tissue factor(TF)in ECV-304 cells;it also enhanced the sensi- tivity of ECV-304 cells to the apoptotic effect of TNF?([16.6?2.5]% vs[25.6?2.3]%,P
8.Study on mechanism of Salvia miltiorrhiza treating cardiovascular disease through auxiliary mechanism elucidation system for Chinese medicine.
Shuai-bing HE ; Bai-xia ZHANG ; Hui-hui WANG ; Yun WANG ; Yan-jiang QIAO
China Journal of Chinese Materia Medica 2015;40(19):3713-3717
Salvia miltiorrhiza is a traditional Chinese medicine (TCM) and is widely used as a clinically medication for its efficiency in treating cardiovascular disease. Due to TCM is a comprehensive system, the mechanism of S. miltiorrhiza treating cardiovascular disease through integrated multiple pathways are still unclear in some aspects. With the rapid progress of bioinformatics and systems biology, network pharmacology is considered as a promising approach toward reveal the underlying complex relationship between an herb and the disease. In order to discover the mechanism of S. miltiorrhiza treating cardiovascular disease systematically, we use the auxiliary mechanism elucidation system for Chinese medicine, built up a molecule interaction network on the active component targets of S. miltiorrhiza and the therapeutic targets of cardiovascular disease to offer an opportunity for deep understanding the mechanism of S. miltiorrhiza treating cardiovascular disease from the perspective of network pharmacology. The results showed that S. miltiorrhiza treating cardiovascular disease through ten pathways as follows: improve lipid metabolism, anti-inflammation, regulate blood pressure, negatively regulates blood coagulation factor and antithrombotic, regulate cell proliferation, anti-stress injury, promoting angiogenesis, inhibited apoptosis, adjust vascular systolic and diastolic, promoting wound repair. The results of this paper provide theoretical guidance for the development of new drugs to treat cardiovascular disease and the discovery of new drugs through component compatibility.
Animals
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Cardiovascular Diseases
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drug therapy
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genetics
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metabolism
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Databases, Factual
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Drugs, Chinese Herbal
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administration & dosage
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chemistry
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Gene Regulatory Networks
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drug effects
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Humans
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Salvia miltiorrhiza
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chemistry
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Signal Transduction
9.Mesenchymal stem cells reduce the irradiation induced lung injury
Ge SHEN ; Fang WANG ; Weijing ZHANG ; Bing LIU ; Shunzong YUAN ; Li WANG ; Yun SHAO
Chinese Journal of Radiological Medicine and Protection 2010;30(6):670-673
Objective To evaluate the role of mesenchymal stem cells (MSCs) derived from mouse bone and embryo dorsal aorta(DA) area in the treatment of irradiation induced lung injury of mouse model. Methods The mice were divided into four groups as normal control group, irradiation group,bone MSCs treatment group and DA MSCs treatment group. Immunohistochemical Analysis of lung tissue was observed after 9 months of treatment. Results Fibrosis and alveolar infiltration were scored in each group. The score for fibrosis and alveolar is 0. 17 in normal control group, 2 in irradiation group, 1 in bone MSCs treat group and 1.38 in DA MSCs treat group. Conclusion The extent of irradiation Induced Lung Injury could be reduced thorough the treatment of MSCs derived from mouse bone and embryos dorsal aorta ( DA ) area.
10.Purification technology of manninotirose in Rehmanniae Radix Praeparata by D-101 microporous adsorption resin.
Xing-xing WANG ; Jun QIAN ; Yun WU ; Bing-peng YAN ; Zhen-zhong WANG ; Wei XIAO
China Journal of Chinese Materia Medica 2015;40(12):2340-2344
This paper was aim to optimize the purification technology of Rehmanniae Radix Praeparata extract with macroporous adsorption resin. With the content of manninotriose as index, the absorptive flow and time were investigated, as well as kinds, amount, flow of eluent. D-101 type macroporous adsorption resin was the best choice for the purification of manninotriose. The optimized parameters were as follows: the content of manninotriose at 161.16-53.72 mg x g(-1), absorption time 240 min, eluting solvent of purified water, volume flow at 1.5 BV x h(-1), and eluant volume at 6 BV. D-101 type macroporous adsorption resin could significantly increase the purity of Rehmanniae Radix Praeparata extract with the advantage of high absorption, remove most part of impurity, and the effect of semi-works production was better.
Adsorption
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Chemical Fractionation
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Chromatography, Liquid
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instrumentation
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methods
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Drugs, Chinese Herbal
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chemistry
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isolation & purification
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Rehmannia
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chemistry
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Resins, Synthetic
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chemistry