2.Purification of Recombinant Fusion Protein Staphylokinase-Hirudin Expressed by Escherichia coli and Analysis of its Dimer
Gen-Shen ZHONG ; Ai-Ping YU ; Ji-De JIN ; Zhong-Hua JIANG ; Zu-Ze WU ;
China Biotechnology 2006;0(02):-
The recombinant fusion protein staphylokinase-hirudin(rSFH) was purified from the high density-fermented engineered E.coli by means of ion-exchange chromatography (IEC) and gel filtration (GF). The purity of rSFH reached to more than 98% determined by RP-HPLC and SDS-PAGE, and the yield was up to 0.7g per liter of fermentation broth. The analysis of homologous dimmer of rSFH appeared during the purification and calculation of the surface hydrophobic area had been carried out by means of hydrophobic chromatography and MALD-TOF. The influence of sodium chloride and temperature on the behavior of rSFH reversible dimerization was analyzed by high performance sized- exclusive chromatography(HPSEC). It is concluded that the hydrophobic interaction played an important role in the reversible dimerization of rSFH.
3.The simultaneous determination of laetrile, paeoniflorin and paeonol in Jingzhi Guizhi Fuling capsule by HPLC.
Zheng-gen LIAO ; Ya LING ; Yan ZHONG ; Qi-neng PING
China Journal of Chinese Materia Medica 2005;30(16):1252-1254
OBJECTIVETo establish an HPLC method for the simultaneous determination of three major bioactive components in Jingzhi Guizhi Fuling capsules namely laetrile, paeoniflorin and paeonol.
METHODA LiChrospher C18 column (4.6 mm x 250 mm, 5 microm) was used. The chromatography was carried out with a stepwise gradient programming. The mobile phase was acetonitrile-water (containing 0.1% phosphorous acid) and the flow rate was 1.0 mL x min.
RESULTThe linear range of laetrile was 12.87-102.94 micron x mL(-1), r = 0.999 9, paeoniflorin 24.84 - 198.7 microg x mL(-1), r = 0.9999 and paeonol 12.57-100.56 microg x mL(-1), r = 0.999 9. The method is accurate with variation less than 1.5 % and recovery more than 95 %.
CONCLUSIONThe method was successfully applied to analyze three major bioactive components in Jingzhi Guizhi Fuling capsules.
Acetophenones ; analysis ; Amygdalin ; analysis ; Benzoates ; analysis ; Bridged-Ring Compounds ; analysis ; Capsules ; Chromatography, High Pressure Liquid ; methods ; Cinnamomum ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; Glucosides ; analysis ; Monoterpenes ; Paeonia ; chemistry ; Plants, Medicinal ; chemistry ; Polyporales ; chemistry ; Reproducibility of Results
4.Expression of osteopontin mRNA and OCT2 mRNA in human gastric carcinoma by tissue microarray and its significance.
Qiao-ying ZHANG ; Gen-you YAO ; Yan-ping FU ; Zhong-sheng ZHAO
Chinese Journal of Pathology 2006;35(1):42-43
Adenocarcinoma
;
metabolism
;
pathology
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Adenocarcinoma, Mucinous
;
metabolism
;
pathology
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Adenocarcinoma, Papillary
;
metabolism
;
pathology
;
Carcinoma, Signet Ring Cell
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metabolism
;
pathology
;
Follow-Up Studies
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Gene Expression Regulation, Neoplastic
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Humans
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Lymphatic Metastasis
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Neoplasm Invasiveness
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Organic Cation Transport Proteins
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biosynthesis
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genetics
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Organic Cation Transporter 2
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Osteopontin
;
Prognosis
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RNA, Messenger
;
biosynthesis
;
genetics
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Sialoglycoproteins
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biosynthesis
;
genetics
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Stomach Neoplasms
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metabolism
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pathology
;
Survival Rate
5.Microsurgical treatment of obstructive azoospermia: a report of 76 cases.
Feng-bin ZHANG ; Zhong-yan LIANG ; Le-jun LI ; Jing-ping LI ; Jing-gen WU ; Fan JIN ; Yong-hong TIAN
National Journal of Andrology 2015;21(3):239-244
OBJECTIVETo investigate the clinical effect of microsurgical vasoepididymostomy and/or vasovasostomy in the treatment of obstructive azoospermia.
METHODSThis study included 76 patients with obstructive azoospermia, 53 treated by bilateral vasoepididymostomy (8 involving the epididymal head, 18 involving the epididymal body, 5 involving the epididymal tail, and 22 involving the epididymal head, body and tail), 14 by unilateral vasoepididymostomy, and the other 9 by unilateral vasoepididymostomy + unilateral vasovasostomy (including cross anastomosis). We followed up the patients for 2 to 16 months for the patency rate, routine semen parameters, and pregnancy outcomes.
RESULTSThe success rate of bilateral vasoepididymostomy, unilateral vasoepididymostomy, and unilateral vasoepididymostomy + unilateral vasovasostomy (including cross anastomosis) were 62.26% (33/53), 35.71% (5/14), and 77.78% (7/9), respectively. The average sperm concentrations in the three groups of patients were (27.9 +/- 5.74), (11.8 +/- 8.33), and (19.9 +/- 7.53) x 10(6)/ml, the average total sperm counts were (65.6 +/- 13.71), (28.0 +/- 15.86), and (69.2 +/- 28.59) x 10(6), and the mean rates of progressively motile sperm were (22.3 +/- 3.18), (11.0 +/- 9.77), and (15.8 +/- 5.05)%, respectively. The success rates of bilateral vasoepididymostomy that involved the epididymal head, body, tail, and all the three parts were 62.5, 72.22, 60, and 54.55%, respectively. Natural pregnancy was achieved in 8 (10.53%) of the total number of cases.
CONCLUSIONMicrosurgery is effective for the treatment obstructive azoospermia. Unilateral vasoepididymostomy + unilateral vasovasostomy is superior to the other procedures, followed by bilateral vasoepididymostomy. Bilateral vasoepididymostomy involving the epididymal body may achieve a slightly better effect than that involving the other epididymal parts.
Adult ; Anastomosis, Surgical ; methods ; Azoospermia ; etiology ; surgery ; Epididymis ; surgery ; Female ; Humans ; Infertility, Male ; surgery ; Male ; Microsurgery ; Pregnancy ; Pregnancy Rate ; Sperm Count ; Treatment Outcome ; Vas Deferens ; surgery ; Vasovasostomy ; methods
6.Fas-FasL and caspase-3 signal transduction pathway and apoptosis of peripheral T lymphocytes in ITP patients.
Yong-Gen ZHONG ; Wei-Ying FENG ; Hong-Qiang LUO ; Jia-Ping FU ; Jie JIN
Chinese Journal of Hematology 2008;29(5):329-332
OBJECTIVETo explore the relationship between Fas-FasL-mediated signal transduction pathway and apoptosis of T lymphocyte subset in ITP patients.
METHODSThe expression rates of membrane Fas, FasL and intracellular activated caspase-3 in peripheral T lymphocyte subset were determined by flow cytometry. T cell subsets with caspase-3 protein expression were detected by Western blot.
RESULTSAs compared with that in healthy control group [(29.4 +/- 8.2)%], the expression rate of membrane Fas on CD4+ T cells was significantly increased in ITP patients [(42.1 +/- 9.5)%] (P < 0.05), however, that on CD8+ T cells was only slightly increased [(9.3 +/- 6.0)% vs (13.4 +/- 5.8)%] with no statistical significance (P > 0.05). The expression rate of FasL on T cell subset in ITP patients was significantly increased (P < 0.05), and that of intracellular activated caspase-3 in T cell subset in ITP patients was notably higher than that in healthy control group (P < 0.05). Western blot analysis showed that the expression of pro-caspase-3 and cleaved-caspase-3 in CD4+ T cells in patients with ITP after treatment were significantly reduced compared with those before treatment (P < 0.05).
CONCLUSIONApoptosis of T lymphocyte subset in ITP patients is accelerated. It is possible that Fas-FasL signal transduction pathway plays an important role in the induction of the apoptosis. The degree of apoptosis of T lymphocytes closely correlates with the disease's activity in ITP patients. Hormone therapy may interfere with Fas-FasL signal transduction pathway of apoptosis.
Adolescent ; Adult ; Apoptosis ; Case-Control Studies ; Caspase 3 ; metabolism ; Fas Ligand Protein ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Purpura, Thrombocytopenic, Idiopathic ; blood ; Signal Transduction ; T-Lymphocytes ; metabolism ; Young Adult ; fas Receptor ; metabolism
7.Expression of mammary serine proteinase inhibitor in oral squamous cell carcinoma and its clinical significance.
Jun LUO ; Lan ZHOU ; Gen-jian ZHENG ; Tong JI ; Lai-ping ZHONG
Chinese Journal of Stomatology 2009;44(10):606-610
OBJECTIVETo investigate the expression of mammary serine proteinase inhibitor(Maspin) in patients with oral squamous cell carcinoma and its relationship with clinicopathological features.
METHODSThe Maspin protein expression was examined in 45 patients with oral squamous cell carcinoma and in non-malignant oral epithelia by immunohistochemical staining. The relationship between the Maspin protein expression and the clinicopathological parameters was statistically analyzed.
RESULTSThe Maspin protein expression in the cancerous tissues of oral squamous cell carcinoma was lower than that in the non-malignant oral epithelia (P = 0.001). Negative correlation was found between the Maspin protein expression in cancerous tissue and the status of lymph node metastasis (P = 0.038). Positive Maspin protein expression indicated negative lymph node metastasis, and negative correlation was also found between the Maspin protein expression and the postoperative metastasis (P = 0.004). Positive correlation was found between the Maspin expression and the patients' survival rate (P = 0.014), patients with positive Maspin protein expression having higher survival rate than those with negative Maspin expression.
CONCLUSIONSMaspin might be useful as a potential prognostic marker for oral squamous cell carcinoma.
Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Female ; Humans ; Male ; Middle Aged ; Mouth Neoplasms ; metabolism ; pathology ; Serine Proteinase Inhibitors ; metabolism ; Serpins ; metabolism
8.Establishment and application of fluorescent quantitative PCR method for detecting human herpes virus type 6.
Xue-jun DONG ; Chang-gen SHI ; Shi-wen QU ; Zhong-min LIU ; Jia-ping FU
Chinese Journal of Experimental and Clinical Virology 2005;19(1):74-76
OBJECTIVETo establish fluorescent quantitative PCR method for detecting human herpes virus type 6 (HHV6).
METHODSAccording to the specific sequence of human herpes virus type 6 genes, the primers and the fluorescent probe (TaqMan) were designed and synthesized. The fragment generated from HHV 6 gene as template was cloned into the pMD18-T vector which was constructed from the pUC 18. And the positive recombinant plasmid was 1:10 diluted and used as standard quantitative template to make the standard curve for sample detection.
RESULTSThe standard curve indicated the linear relationship between Ct (cycle threshold) and template concentration. The clinical samples from 135 cases were detected by this system, 16 cases among 135 were positive.
CONCLUSIONThe fluorescent quantitative PCR method for the detection of human herpes virus type 6 is simple and accurate, and this method may be helpful to clinical diagnosis.
Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; DNA Primers ; DNA Probes ; DNA, Viral ; genetics ; Female ; Herpesvirus 6, Human ; genetics ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; methods ; Purpura, Thrombocytopenic, Idiopathic ; diagnosis ; virology ; Reproducibility of Results ; Roseolovirus Infections ; diagnosis ; virology ; Sensitivity and Specificity ; Young Adult
9.Inheritance and innovation of traditional Chinese medicinal authentication.
Zhong-zhen ZHAO ; Hu-biao CHEN ; Pei-gen XIAO ; Ping GUO ; Zhi-tao LIANG ; Fanny HUNG ; Lai-lai WONG ; Eric BRAND ; Jing LIU
China Journal of Chinese Materia Medica 2015;40(17):3385-3390
Chinese medicinal authentication is fundamental for the standardization and globalization of Chinese medicine. The discipline of authentication addresses difficult issues that have remained unresolved for thousands of years, and is essential for preserving safety. Chinese medicinal authentication has both scientific and traditional cultural connotations; the use of scientific methods to elucidate traditional experience-based differentiation carries the legacy of Chinese medicine forward, and offers immediate practical significance and long-term scientific value. In this paper, a path of inheritance and innovation is explored through the scientific exposition of Chinese medicinal authentication, featuring a review of specialized publications, the establishment of a Chinese medicine specimen center and Chinese medicinal image databases, the expansion of authentication technologies, and the formation of a cultural project dedicated to the Compedium of Materia Medica.
Drug Contamination
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prevention & control
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Drugs, Chinese Herbal
;
chemistry
;
standards
;
Humans
;
Materia Medica
;
chemistry
;
standards
;
Medicine, Chinese Traditional
;
standards
;
Reference Standards
10.0258-879X(2001)04-0306-02 Effect of dobutamine on dog myocardium integrated backscatter
Wen-Feng XIONG ; Bao-Zhen ZHAO ; Shi-Ping LU ; Jun QIAN ; Gen-Bao ZHAO ; Bo YANG ; Zhong WANG
Academic Journal of Second Military Medical University 2001;22(4):306-307
Objective: To understand the effect of dobutamine on the integrated backscatter of myocardium. Methods: Nineteen open-chest dogs were detected by HP Sonos 5500 with S8 transducer, and the WT, AIBS and CVIB were measured before and after the dobutamine stress. Results: During the dobutamine stress, following the increase of WT, the transmural, subepicardial and subendocardial CVIB all increased. And the increase of subepicardial CVIB was greater than that of subendocardial CVIB, leading to the decrease of TGI. But the AIBS didn't change significantly in the whole process. Conclusion: Dobutamine can enhance myocardium CVIB but has no obvious infleunce on AIBS.