1.Determination of Residual of 4 Kinds Organic Solvents in Ecabet Sodium Crude Drug by Capillary GC
Xue DI ; Yanhua LIU ; Haibo WANG ; Hui ZHANG ; Tianmin WANG
China Pharmacy 2015;(30):4286-4287,4288
OBJECTIVE:To establish a method for the determination of residuals of petroleum ether,ethanol,xylene and ace-tic acid in ecabet sodium crude drug. METHODS:Capillary GC was performed on the column of PGE-20M capillary column at the flow rate of 1.7 ml/min,detector was hydrogen flame ionization detector,carrier gas was nitrogen with high purity,column temper-ature was 45 ℃,maintaining 4 min,it increased to 80 ℃ with rate of 10 ℃/min,then increased to 135 ℃ with rate of 30 ℃/min,maintaining 3 min,the injection mode was direct injection,inlet temperature was 250 ℃,and the volume injection was 1.0 μl. RESULTS:The mass concentration was 0.050-1.952 g/L for petroleum ether,0.050-1.941 g/L for ethanol,0.024-0.948 g/L for xy-lene and 0.050-1.947g/L for acetic acid(r=0.999 1-0.999 7);RSDs of precision,stability and reproducibility tests were no more than 1.7%;recoveries were 99.3%-101.0%(RSD=0.7%,n=9),102.3%-103.7%(RSD=0.4%,n=9),101.2%-102.1%(RSD=0.3%,n=9) and 100.3%-102.2%(RSD=0.6%,n=9),respectively. CONCLUSIONS:The method is simple and accurate,and can be used for the control of residual of organic solvents in ecabet sodium crude drug.
2.Local radioimmunotherapy of breast cancer-bearing nude mice using anti-CEA monoclonal antibody C50 labeled with ~(131)I
Weiqin CHANG ; Tianmin XU ; Ying JIANG ; Xinming WANG ; Jitong ZHOU
Journal of Jilin University(Medicine Edition) 2006;0(05):-
Objective To investigate and evaluate the curative effect of radioimmunological targeting drug on nude mice bearing breast cancer. Methods The anti-CEA monoclonal antibody C50 was combined with ~ 131 I to produce radioimmunological targeting drug. 16 nude mice inoculated subcutaneously with breast cancer cell MCF-7 with tumor diameter about 0.5 cm were randomly into 4 groups(n=4): group Ⅰ, injected in part with ~ 131 I-C50 18.5 MBq; group Ⅱ, injected in part with ~ 131 I-C50 3.7 MBq; group Ⅲ, injected in part with ~ 131 I-mIgG 18.5 MBq; group Ⅳ, injected in part with C50 0.75 ?g. The size of tumor volume and inhibitory rate (IR) after treatment for six weeks were calculated and compared with the control group. Results The tumor volume and curves for tumor growth and tumor weight had significant differences between group Ⅰ and the group Ⅲ as well as group Ⅱ (P0.05). Conclusion Anti-CEA monoclonal antibody C50 labeled with radionuclide ~ 131 I could inhibit the growth of the tumor when given locally. ~ 131 I-C50 has a potential value of clinical application
3.Construction of eukaryotic expressing vector pEGFP-N1/PDGF-A for transducting Dermis-derived mesenchymal stem cells
Guohe YAN ; Yongping SU ; Junping WANG ; Daijie WANG ; Guoping AI ; Fengchao WANG ; Xinze RAN ; Tianmin CHENG
Journal of Third Military Medical University 2003;0(20):-
Objective To clone platelet-derived growth factor A chain (PDGF-A) gene and insert PDGF-A gene into. Enhanced green fluorescent protein (EGFP) vector and then transformed into dermis-drived mesenchymal stem cells (DMSCs). Methods cDNA clones encoding human PDGF-A gene were isolated from a human hepatoma cell line mRNA by reverse transcription-polymerase chain reaction (RT-PCR). The PCR amplified fragment of PDGF-A gene was cloned into pMD18-T vector. The eukaryotic expression vector pEGFP-N1/PDGF-A was constructed by subcolone PDGF-A gene into pEGFP-N1 vector. PDGF-A gene was transfected into DMSCs with the help of Fugene 6 transfection reagent. Results Full cDNA sequence encoding human PDGF-A gene had been cloned, which sequence was consistent with the reported sequence in GenBank by sequence assaying. Conclusion cDNA sequence encoding human PDGF-A gene was successfully cloned into pEGFP-N1. The transient expression of PDGF-A gene in DMSCs has been realized.
4.Cultivation of medical students' professional information literacy competence in combination with the reform on medial education and teaching
Tianmin SUN ; Bochun YAO ; Shunhua XIONG ; Jun WANG ; Jinyong WANG ; Wenchun LI
Chinese Journal of Medical Education Research 2011;10(9):1056-1058
It has been a major topic for the reform on higher medical education and teaching to cultivate medical students' professional information literacy so asto make them equipped with the information competence required for disease control or medical research.In this thesis,the concept of professional information literacy for medical students was introduced,and by analyzing the relationship of information literacy education and medical education modes,countermeasures and methods of cultivating medical students' professional information literacy by combining the reform on medical education and teaching were put forward.
5.Cloning and Expression of the CryIVD Gene of Bacillus thuringiensis subsp. israelensis and its Mosquito Larvicidal Activity
Xin ZHANG ; Xiangping LIU ; Ge YAN ; Tianmin ZHEN ; Xinguo WANG ; Chuanhong SUN ; Huaiwei WANG ;
Chinese Journal of Parasitology and Parasitic Diseases 1997;0(05):-
Objective To clone and express Bacillus thuringiensis subsp. isrealensis(B.t.i.) crystal protein CryIVD gene and determine its mosquito larvicidal activity. Methods The gene encoding CryIVD (2.0 kb or so) was amplified by PCR, the amplified fragment was inserted into E.coli plasmid pUC18 to construct the recombinant cloning and expression vector pUC18 CryIVD, which was named pUC18 1. The ligation was transformed into competent E.coli DH 5? and the recombinant vector pUC18 1 was confirmed by restriction enzyme digestion and DNA sequencing. After being inducted by IPTG, the expression of CryIVD gene in positive clone was detected by SDS PAGE and the mosquito larvicidal activity of CryIVD was also determined by standard bioassay. Results The results showed that the CryIVD gene was successfully cloned and expressed in E.coli DH 5? . Mosquito larvicidal activity of engineered E.coli (LC 50 ) to Cx.pipiens pallens and Ae.albopictus Ⅱ-Ⅲ instar larvae was 2.38?10 6 cells/ml and 1.6?10 7cells/ml respectively. Conclusion The CryIVD gene was successfully cloned and expressed, and a high mosquito larvicidal activity was observed.
6.Effect of compound huangjing oral liquid on myocardial myosin heavy-chain in rats with heart failure
Jinshui CHEN ; Tianmin WU ; Shengyuan LIN ; Jian DU ; Kegui WU ; Huajun WANG ; Xiaoming CHEN
Chinese Journal of Tissue Engineering Research 2005;9(39):190-192
BACKGROUND: Adriamycin is anthracycline-based drugs of anti-cancer and inhibits many malignant tumors. But due to the large toxicity, it will induce dose-dependent cardiac toxicity, resulting in heart failure in severe case. Compound huangjing oral lipid is against the injury of free radial and is expected to be applied as an assistant therapy for heart failure.OBJECTIVE: To probe into the therapeutic effect of compound huangjing oral lipid and its mechanism on heart failure.DESIGN: Randomized controlled observation was designed.SETTING: Department of Traditional Chinese Medicine , First Affiliated Hospital of Fujian University of Medical Science, Fujian College of Traditional Chinese Medicine.MATERIALS: The experiment was performed in Fujian Research Institute of Hypertension from August 2000 to May 2001, in which, 66 rats were employed and randomized into 6 groups, 11 rats in each one.METHODS: In normal group, physiological saline of equal volume was injected abdominally. In adriamycin group, adriamycin 1mg/kg was injected abdominally on the 2nd and 4th days after experiment, 2 mg/kg was injected on the 6th and 8th days, 3 mg/kg was on the 10th and 12th days and 4 mg/kg was on the 14th and 16th days. The dose was accumulated up to 20 mg/kg in 16 days. In adriamycin+compound huangjing oral liquid 2 mL (small-dose group), adriamycin +compound huangjing oral liquid 4 mL (moderate-dose group) and adriamycin+compound huangjing oral liquid 6ml (large-dose group), the oral lipid of various doses was applied for gastric perfusion everyday successively from the beginning of experiment, in which, the dose of adriamycin was same as adriamycin group. In adriamycin+tebonin group (tebonin group), tebonin 450 mg/kg was administrated once every two days, totally for 8 days, in which, the dose of adriamycin was same as adriamycin group.MAIN OUTCOME MEASURES: To observe the changes of Left ventricular weight and body weight (LVW/BW), α-MHC (myosin heavy-chain)and β-MHC.RESULTS: In the whole experiment, of 66 experimental animals, 5 rats in adriamycin group, 4 rats in small-dose group, 2 rats in moderate-dose group, 3 rats in large-dose group were died from obvious congestive heart failure, finally, 47 rats entered result analysis. Compared with normal group, in adriamycin group, α-MHC was reduced by 20.88% (P < 0.01), β-MHC was increased by 50.93% (P < 0.01) and LVW/BW was increased by 33.83% (P < 0.01). After medication, myocardial β-MHC was transformed to α-MHC; compared with adriamycin group, α-MHC in every medical group was increased (P < 0.01), β-MHC was decreased (P < 0.01) and LVW/BW was decreased of different degrees (P < 0.01 or P < 0.05); among the above-changes, the results in moderate group were the best.CONCLUSION: Compound huangjing oral liquid alleviates toxicity of heart failure induced by adriamycin, probably due to the dose-dependence improvement of the oral liquid in myocardial α-MHC transformation.
7.Randomized controlled study of flexible ureteroscopic lithotripsy and mini-percutaneous nephrolithotomy for treating upper ureteral calculi
Tianmin LI ; Luofu WANG ; Weihua LAN ; Jianxin JIANG ; Jianghua WAN ; Zhilin NIE ; Jun JIANG ; Keqin ZHANG
Chongqing Medicine 2014;(11):1296-1298
Objective To compare the safety and efficacy of flexible ureteroscopic lithotripsy (F-URS) and mini-percutaneous nephrolithotomy(MPCNL) for treating upper ureteral calculi by the randomized controlled clinical trial .Methods 120 cases of up-per ureteral calculi treated in this hospital from May to September 2013 were randomized into the F-URS group and the MPCNL group with 60 cases in each group .The postoperative clinical curative effects were compared between the two groups .Results The initial stone clearance rates of the F-URS group and the MPCNL group were 68 .33% and 95 .00% respectively ,and the difference was statistically significant (P< 0 .05) .The clearance rate of the two groups after postoperative 3 months were 96 .67% and 98 .33% respectively ,and the difference between the two groups had no statistical significance (P>0 .05) .The operating time of the F-URS group and the MPCNL group was (35 .33 ± 11 .57)min and (53 .75 ± 17 .31)min respectively ,the average hospital stay after operation was(3 .05 ± 0 .62)d and (5 .43 ± 1 .84)d respectively ,and the average visual analogue scale(VAS) score on first postoper-ative day was 1 .70 ± 1 .37 and 3 .68 ± 1 .57 respectively .These indicators had statistically significant differences between the two groups(P<0 .05) .The average dropped amount of hemoglobin and the occurrence rate of complications after operation had no sta-tistically significant differences between the F-URS group and the MPCNL group(P>0 .05) .Conclusion For upper ureteral calculi with surgical indication ,the effect of F-URS is equivalence to MPCNL in the aspect of the stone clearance rate ,but F-URS has high-er security and shorter postoperative hospital stay .In the department with the condition to conduct F-URS ,F-URS should be ranked as the preferred treatment method for upper ureteral calculi .
8.MPS in diagnosing and treating gastrointestinal submucosal protrusive lesions
Junbo QIAN ; Huasheng LU ; Tianmin CHEN ; Yamin WANG ; Zhenming GE ; Yuning LI
Chinese Journal of Digestive Endoscopy 1996;0(06):-
Objective To evaluate the value of miniprobe sonography (MPS) in diagnosing gastrointestinal submucosal protrusive lesions and selecting the indicated manual for treatment. Methods According to the sizes, properties, and depth of SMTs in the gastrointestinal tract detected by the MPS, different methods of resection were performed. Results Of 24 cases, 11 SMTs lying in submucosa under 2cm in diameter (2 benign gastrointestinal stromal tumors, 3 lipomas, 5 cysts, 1 granular cell tumor) were attempted with EMR or argon plasma coagulation (APC) ; there were no complications of hemorrhage or perforation. Thirteen SMTs lying in muscularis propria or with size of SMTs above 2 cm in diameter (4 malignant GISTs, 6 benign GISTs, 1 lipoma, 2 aberrant pancreas) were performed by surgical resection. Preoperative diagnoses of SMTs by MPS were consentient with their histological diagnoses. Conclusion MPS may detect the size, property, and depth of SMTs in the gastrointestinal tract and is helpful in selecting indicated cases for endoscopic resection. Endoscopic therapy of SMTs lying in mucosa or submucosa under 2cm in diameter is a safe and effective procedure.
9.Morphological study on the growth of human amniotic membrane loaded with porcine bone marrow-derived mesenchymal stem cells
Guohe YAN ; Guoping AI ; Daijie WANG ; Zhongmin ZOU ; Xinze RAN ; Junping WANG ; Rong LI ; Yongping SU ; Tianmin CHENG
Chinese Journal of Tissue Engineering Research 2007;11(15):2985-2989
BACKGROUND: Human amniotic membrane (HAM) contains various ingredents such as collagen, glycoprotein,proteoglycan, integrin and laminated body, and so on, and expresses many kinds of growth factors and mRNA-associated proteins. And these ingredents can supply abundant nutriments for cellular proliferation and differentiation, and benefit cells to grow and propagate. Whether or not HAM can load porcine bone marrow-derived mesenchymal stem cells (BMSCs) to well grow on it deserves to be further investigated.OBJECTIVE: To set up a method of tissue engineering of human amniotic membrane loading porcine BMSCs and observe the morphological characteristics of growth and proliferation of BMSCs seeded on HAM.DESIGN: Randomized controlled observation.SETTING: State Key Laboratory of Trauma, Burn and Combined Injury, General Institute of Combined Injuries, Academy of Preventive Medicine, Third Military Medical University of Chinese PLA.MATERIALS: This experiment was carried out in the State Key Laboratory of Trauma, Burn and Combined Injury,General Institute of Combined Injuries, Academy of Preventive Medicine, Third Military Medical University of Chinese PLA between January and November 2003. Three Guizhou minipigs of either gender, aged 2 to 3 months, weighing from 6 to 8 kg, were provided by the Experimental Animal Center, Third Military Medical University of Chinese PLA. Main reagent:ISCOVE'S modified DULBECCO'S medium (IMDM) culture medium (Hyclone, USA); high-quality fetal bovine serum PAA (Germany); haematoxylin (China); Eosin B (Sigma, USA) and OCT embedding medium (USA). Main instruments: BX51 stereoscopic fluorescence microscope (Olympus, JaPan); IX70 inverted fluorescence microscope (Olympus, Japan);cryostat (2700-Frigcut, Germany); myeloid puncture needle (Jiangsu); superclean bench (Sujing Bloc Antai Company);CO2 constant-temperature incubator (QUEUE, USA).METHODS: HAM was prepared as previously described. The BMSCs of Guizhou minipigs isolated and cultured according to method described previously were primarily cultured and passaged, then they were inoculated to the stromal surface of HAM at different densities (0.84×105 cells/cm2,1.54×105 cells/cm2,2.75×105 cells/cm2); The growth and proliferation of BMSCs of different densities were observed under an inverted microscope and scanning electron microscope; BMSCs of the second or the third passages were inoculated on HAM held with tissue-holding device at a density of 1.54×105 cells/cm2, and they were cultured for 18 days at most. The HAM was daily rolled, sliced and stained by HE for observing the growth of BMSCs loaded on HAM under the light, scanning and transmission electron microscopes.MAIN OUTCOME MEASURES: The growth of BMSCs on HAM was examined at different densities and different time points.RESULTS: ① Comparison of growths of BMSCs promoted by different densities of HAM: BMSCs,which were planted on HAM at the density of 0.84×105 cells/cm2 were irregular and scattered under an invert microscope. Distances between BMSCs were biggish. BMSCs seeded on HAM at the density of 1.54×105 cells/cm2 were regular in arrangement and moderate in density, with clear cell outline and good cell activity before 24 hours, and seeded at the density of 2.75×105 cells/cm2 were congested with many nonattached cells and the longer the growing time of the cells was, the more the cellular debris were observed. BMSCs,which were planted on HAM at the density of 0.84×105 cells/cm2 under the scanning electron microscope, scatted on HAM presented in shapes of irregular, long, thin and flat polygon. Their membrane protuberances presented in shapes of thick and thin, and the distances between cells were biggish. BMSCs,which were planted on HAM at the density of 1.54×105 cells/cm2 have similar appearance of their bodies and membrane protuberances, and the membrane protuberances were more compared with the BMSCs planted at the density of 0.84×105 cells/cm2. Their membrane protuberances intercrossed each other, and the margin of some BMSCs overlapped each other. BMSCs planted at the density of 2.75×105 cells/cm2, arraved on HAM crowdedly and overlappedly with many debris. Their membrane protuberances were not obviously. The margin of some BMSCs was overlapped.② Comparisonof growths of BMSCs promoted by HAM at different time points: Under the inverted microscope, the BMSCs adhered quickly to HAM after being incubated for about 30 minutes. All of BMSCs adhered to HAM within 24 hours, and formed monolayer on it within 48 hours, and grew densely on HAM after being cultured for 4 to18 days. Under the light and electron microscopes, HE results revealed that BMSCs adhered tightly and grew on HAM in different arrays, such as emitting, whirlpool or parallel,and their nuclei located in middle, dense in staining, were big and clear. The shapes of BMSCs were comparatively consistent on HAM. HAM loaded with BMSCs grew 4 days, and BMSCs covered HAM completely. The densities of BMSCs on HAM were suitable, and their bodies were large, and presented irregular, long,thin and flat polygon under the scanning electron microscope. The margin of some BMSCs overlapped each other. The protuberances of cellular membrane of BMSCs were abundant in the shapes of thick and thin. Some protuberances intercrossed each other in the shape of net. BMSCs adhered tightly to HAM through these protuberances. HAM loading BMSCs grow 4 days; most of BMSCs grew on HAM in double layers with the shapes of cambiform under the transmission electron microscope, Their nucleoli were clear. The protuberances of cellular membrane of BMSCs, which situated at two sides of nuclei and overlapped each other, were long. Most of chromatins of BMSCs were autosome.Abundant organell such as rough endoplasmic reticulum (RER),mitochondria could be observed in BMSCs.CONCLUSION:HAM is able to promote the proliferation of BMSCs significantly. BMSCs may be cultured on HAM ex vivo.HAM is a good carrier of BMSCs.
10.Human amniotic membrane loaded with porcine keratinocytes for constructing epidermal substitute of skin
Guohe YAN ; Yongping SU ; Feng WANG ; Guoping AI ; Tianmin CHENG ; Huaien ZHENG ; Xinze RAN ; Hong XIAO ; Chongfu TAN
Chinese Journal of Tissue Engineering Research 2005;9(22):245-247
BACKGROUND: As a kind of semitransparent membrane, human amniotic membrance contains many kinds of nutrients, which is a good biological material loaded with keratinocytes.OBJECTIVE: To construct epidermal substitute of the skin from human amniotic membrane loaded with porcine keratinocytes and examine the morphological characteristics of the growth and proliferation of keratinocytes seeded on human amniotic membrane.DESIGN: Single sample study and repetitive measured observation based on the cells.SETTING: Institute of Combined Injuries of Chinese PLA, Academy of Preventive Medicine, Third Military Medical University of Chinese PLA.MATERIALS: The experiment was completed in the State Key Laboratory of Trauma, Burn and Combined Injury and Institute of Combined Injuries of Chinese PLA, Academy of Preventive Medicine, Third Military Medical University of Chinese PLA from January to November 2001. Porcine keratinocytes was collected from Guizhou minipigs aged 3 weeks.METHODS: The primarily cultured keratinocytes of Guizhou minipigs were subcuhured, expanded and bred on the stroma surface of human amniotic membrance at the density of 1.63 × 105/cm2. The growth and proliferation of keratinocytes were observed under inverted microscope every day. From the 3rd day and the 15th day after being cultured, the growth of keratinocytes on human amniotic membrane was examined under light microscope and electron microscope.MAIN OUTCOME MEASURES: The growth of keratinocytes on human amniotic membrane was examined RESULTS: Keratinocytes evidently adhered to the stroma surface of human amniotic membrane about 30 minutes after being cultured, which was observed under inverted microscope. Most keratinocytes grew and adhered to the stroma surface of human amniotic membrane within 24 hours. Monolayer of keratinocytes formed and completely covered human amniotic membrane within 3 days. It was observed under the light microscope that the monolayer of keratinocytes adhered to human amniotic membrane and arrayed tightly. The keratinocytes presented in the shape of polygon, and plasmalemmas of keratinocytes formed many pseudopods under the observation with scanning electron microscope. Keratinocytes adhered to human amniotic membrane well and with many keratinofilaments in them under the observation with transmission electron microscope. Keratinocytes arrayed on human amniotic membrane densely with many cellular debris and some keratinocytes formed cavitations in them due to aging after growth for 15 days under the observation with inverted microscope.CONCLUSION: Human amitotic membrane is a good carrier of keratinocytes cultured on it in vitro, and is able to promote the proliferation of keratinocytes significantly. However, when keratinocytes were loaded on the human amniotic membrane for 15 days, some keratinocytes formed cavitations in them due to aging.