2.Construction of subtracted cDNA libraries of gastrocarcinoma and normal tissue with suppression subtractive hybridization and their quality analysis
Lan-jun, WU ; Bing-zhi, MAO ; Sheng-qi, WANG
Bulletin of The Academy of Military Medical Sciences 2001;25(1):9-12,53
Objective: To construct subtracted cDNA libraries of stomach tumors and normal stomach tissue using suppression subtractive hybridization(SSH).Methods: cDNA Library subtraction was performed using the protocol described in the Clontech PCR-Select cDNA Subtraction Kit. cDNA was synthesized from 2 μg of poly A+RNA from the tumor and normal tissues using AMV reverse transcriptase. The tester and driver cDNAs were digested with RsaⅠ, a four-base-cutting restriction enzyme that yields blunt ends. The tester cDNA was then subdivided into two portions, and each was ligated with different cDNA adaptor. Two hybridizations were performed. In the first, an excess of driver was added to each sample of tester. Hybridization kinetics led to equalization and enrichment of differentially expressed sequences. During the second hybridization, the two primary hybridization samples were mixed together without denaturing and thus the templates were generated from differentially expressed sequences for PCR amplification. Using suppression PCR, only differentially expressed sequences were amplified exponentially and after second PCR amplification the background was reduced and differentially expressed sequences were further enriched. The cDNAs were then directly inserted into a T/A cloning vector to generate a stomach tumor-specific subtracted cDNA library. Results: The amplified library contained 800 positive clones. Plasmid inserts were PCR amplified and showed 250-700 bp inserts. Conclusions: The successfully constructed subtracted cDNA library of gastrocarcinoma and normal tissue enables us to compare two populations of mRNA and obtain clones of genes that expressed in one population but not in the other.The characterization of these genes will allow them to be exploited for their diagnostic and therapeutic potentials.
3.Quality evaluation and stability investigation of asarone submicro emulsion injection.
Hong-Jia LI ; Xiu-Jun LAI ; Wei LI ; Ting CHU ; Hui JIN ; Sheng-Jun MAO
China Journal of Chinese Materia Medica 2014;39(20):3945-3949
The content of the asarone submicro emulsion injection was determind by HPLC method, and thereby a quality evaluation method was established based on indexes of pH value, particle size, peroxide value, methoxy aniline values, free fatty acid, lysophosphatidylcholine, visible foreign substances, insoluble particle, sterility, bacterial endotoxin and impurities, etc. The results showed that the injection exhibited uniform physical appearance and all the products were in milkwhite liquid. The content of the three batches products were respectively 102.9%, 100.8%, 97.70% of the labeled amount, with mean particle size of 210-250 nm, and other indexes all met with the standards. The reserved samples showed no obvious change in terms of detection indexes and indicated good stability after the accelerated stability test and long-term stability for 12 months. The quality evaluation method established in this study could be applied to quality control and stability investigation of asarone submicron emulsion injection, which laid a basis for further clinical research and application.
Anisoles
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chemistry
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Chromatography, High Pressure Liquid
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Drug Stability
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Drugs, Chinese Herbal
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chemistry
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Emulsions
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chemistry
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Particle Size
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Quality Control
4.The significance of serum concentrations of transforming growth factor-β1 and bone morphogenetic protein-4 in patients with rheumatoid arthritis and interstitial lung disease
Jun SHENG ; Tihong SHAO ; Dan XUAN ; Tongjun MAO ; Zhi LI ; Jinming LU ; Liang XU
Chinese Journal of Rheumatology 2012;16(8):545-548
Objective To compare the serum levels of transforming growth factor (TGF)-beta 1,bone morphogenetic protein (BMP)-4 in patients with rheumatoid arthritis (RA) and RA with interstitial lung disease (RA-ILD).Methods Twenty-eight RA-ILD patients,32 patients with RA but without ILD and 20 normal controls were enrolled.The RA-ILD group was further divided into early group and late group.All the observed subjects were analyzed using enzyme linked immunosorbent assay (ELISA) for the determination of serum TGF-β1 and BMP-4 levels.The relationship between the serum levels of TGF-β1,BMP-4 and laboratory examinations were investigated.Comparisions between groups were tested by one-way ANOVA analysis and ttest.Correlation of indexs were observed by Spearman method.Results Patients in the RA-ILD group were older than RA group in disease onset age,in addition,patients with ILD had better joint function and higher serum rheumatoid factor titers.The occurrence time of interstitial lung disease was 2-6 years after the onset of arthritis,with an average time of (3.0±1.2) years.The TGF-β1 levels in the RA-ILD group were slightly higher,but not statistically significant than other groups (P>0.05).The TGF-β1 serum levels in the early RA-ILD patients were significantly increased than those of the late RA-ILD group and the RA group.BMP-4 levels in patients with RA-ILD group were less than RA without ILD group and healthy control group,and the difference was statistically significant (P<0.05).BMP-4 level in early RA-ILD group was significantly decreased than those of the late RA-ILD group and RA group.No correlation between the serum BMP-4,TGF-β1 level (P>0.05) no assay result correlated with laboratory parameters including ESR,CRP,RF and anti-CCP antibodies (P>0.05).Conclusion TGF-β1 serum levels are increased and BMP-4 levels are decreased in early RA-ILD patients.The serum levels of TGF-β1 and BMP-4 may be indicatior for asymptomatic ILD and reflect disease progression.
5.Study on the professional assessment standards of clinical pharmacy
Yan LIU ; Xiao-feng LIU ; Jun-hao JIANG ; Qin-geng LI ; Mao-sheng YANG ;
Chinese Journal of Medical Education Research 2011;10(8):939-941
Professional assessment in Chinese Higher Education has made great progress in three stages: the sporadic practice, trial and promotion. The authors present several comments on the characteristics and the professional assessment standards of clinical pharmacy in China, and focus on the scientific system of professional assessment.
6.Development of biphasic drug-loading lipid emulsion of Salvia miltiorrhiza and its quality evaluation.
Yin-Yan WANG ; Xi LI ; Xiu-Jun LAI ; Wei LI ; Ya-Jing YANG ; Ting CHU ; Sheng-Jun MAO
China Journal of Chinese Materia Medica 2014;39(19):3748-3752
The feasibility of simultaneously loading both liposoluble and water-soluble components of Salvia miltiorrhiza in emulsion was discussed, in order to provide new ideas in comprehensive application of effective components in S. miltiorrhiza in terms of technology of pharmaceutics. With tanshinone II (A) and salvianolic acid B as raw materials, soybean phospholipid and poloxamer 188 as emulsifiers, and glycerin as isoosmotic regulator, the central composite design-response surface method was employed to optimize the prescription. The coarse emulsion was prepared with the high-speed shearing method and then homogenized in the high pressure homogenizer. The biphasic drug-loading intravenous emulsion was prepared to investigate its pharmaceutical properties and stability. The prepared emulsion is orange-yellow, with the average diameter of 241 nm and Zeta potential of -35.3 mV. Specifically, the drug loading capacity of tanshinone II (A) and salvianolic acid B were 0.5 g x L(-1) and 1 g x L(-1), respectively, with a good stability among long-term retention samples. According to the results, the prepared emulsion could load liposoluble tanshinone II (A) and water-soluble salvianolic acid B simultaneously, which lays a pharmaceutical foundation for giving full play to the efficacy of S. miltiorrhiza.
Chemistry, Pharmaceutical
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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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Emulsions
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chemistry
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Quality Control
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Salvia miltiorrhiza
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chemistry
7.Analysis of clinical features of 56 cases with Terrien's marginal degeneration
Zhen-Jun, FANG ; Kang-Sheng, LUO ; Zhi-Yong, WU ; Wei, WEI ; Mao-Jun, YANG ; Xiao-Li, ZHAO
International Eye Science 2016;16(8):1548-1550
?AIM:To analyze the clinical features of Terrien marginal degeneration ( TMD) .?METHODS:Fifty-six patients(90 eyes) admitted in our hospital from January 2013 to January 2015 were selected as observation group.At the same period, 56 healthy (88 eyes) for corneal examination were as control group to analyze the clinical features of TMD. With immunohistochemistry and enzyme linked immunoassay method ( ELISA), the levels of HLA-DR, HLA-DQ and tumor necrosis factor-α( TNF-α) in blood samples of TMD patients were tested.?RESULTS:The transparent degree of the eye, lipid deposition in TMD patients with early, advanced, swelling and hole-wearing period were significantly different (χ2=10.85,χ2=65.32, P<0.05).Astigmatism in TMD patients with early and advanced, swelling and hole -wearing period were significantly different (P<0.05).The levels of HLA-DR, HLA-DQ and TNF-αin blood samples between the two groups were significantly different ( t=45.326, t=23.564, t=19.86, P<0.05).?CONCLUSION: Terrien's marginal degenerative is an inflammatory disease characterized by increased levels of TNF-α, HLA-DQ, and HLA-DR in peripheral blood, decreased corneal transparency, astigmatism and lipid deposition.This research provides experiment evidence for the mechanism of TMD.
8.Application of side to side anastomosis between the lesser curvature of stomach and jejunum in laparoscopic Roux-en-Y gastric bypass.
Ri-xing BAI ; You-guo LI ; Jun XU ; Zhi-qiang ZHONG ; Wen-mao YAN ; Hui-sheng YUAN ; Mao-min SONG
Chinese Journal of Gastrointestinal Surgery 2013;16(7):648-650
OBJECTIVETo evaluate the application of side-to-side anastomosis of the lesser curvature of stomach and jejunum in laparoscopic Roux-en-Y gastric bypass (LRYGB).
METHODSClinical data of 29 patients with type 2 diabetes mellitus (T2DM) undergoing side to side anastomosis of the lesser curvature of stomach and jejunum in LRYGB from May 2012 to November 2012 in Department of General Surgery, Beijing Tiantan Hospital, Capital Medical University were analyzed retrospectively.
RESULTSAll the procedures were successfully completed without conversion to laparotomy. The side-to-side anastomosis of the lesser curvature of stomach and jejunum avoided the laparoscopic suture. No gastrojejunostomy anastomotic bleeding, fistula, obstruction and other complications occurred after operation and no complications of gastrojejunostomy anastomosis were found during a follow up of 1 to 7 months.
CONCLUSIONSSide-to-side anastomosis of the lesser curvature of stomach and jejunum in LRYGB can manipulate the size of anastomosis accurately and avoid the laparoscopic suturing. It is simple and easy to learn.
Adult ; Aged ; Diabetes Mellitus, Type 2 ; surgery ; Female ; Follow-Up Studies ; Gastric Bypass ; methods ; Humans ; Jejunum ; surgery ; Laparoscopy ; methods ; Male ; Middle Aged ; Retrospective Studies ; Stomach ; surgery ; Treatment Outcome