1.New porous beta-tricalcium phosphate as a scaffold for bone tissue engineering
Yong LIU ; Guoxian PEI ; Shan JIANG ; Gaohong REN
Chinese Journal of Tissue Engineering Research 2008;12(23):4563-4567
BACKGROUND: New porous β-tricalcium phosphate (TCP) was made by appropriate prescription and unique technology, with a porosity of (75±10)%, spheroidal hole>80%, micropore<20%, interlink rate between the holes of 100% and mechanical strength>2MPa.OBJECTIVE: To assess the application outcome of the new porous β-TCP as a scaffold for bone tissue engineering.DESIGN, TIME AND SETTING: The control experiment was performed at the Laboratory of Tissue Engineering of Southern Medical University, China from July 2005 to March 2006.MATERIALS: Twelve 6-month New Zealand rabbits were used to create 1.5cm large bone and periosteum defects of the left radial bone. Porous β-TCP was purchased from bio-lu, France.METHODS: Osteoblasts differentiated from rabbit bone marrow mesenchymal stem cells (BMSCs) were co-cultured with porous β-TCP. Inverted phase contrast microscope and scanning electron microscope were used to observe the growth of BMSCs. MTT assay was employed to assess cell proliferation and compatibility. Cytotoxicity was detected by analyzing the effects of different concentrations of porous β-TCP leaching liquor on cell proliferation.MAIN OUTCOME MEASURES: Cell compatibility and cytotoxicity of β-TCP were measured. The status of bone defect repair was appraised by histology, radionuclide bone scan and X-ray at 2, 6, 12 weeks after surgery.RESULTS: The new porous β-TCP had good cell adhesion and its cytotoxicity was in 0 grade. Histology, imageology and radionuclide bone scan showed the new porous β-TCP could repair large radial bone defect in rabbits. At the same time, its degradation rate was accordance with bone formation rate in vivo.CONCLUSION: The new porous β-TCP with a good compatibility is a good scaffold for bone tissue engineering, and obtains good outcomes in repairing large bone defect of rabbit radial bone.
2.Effects and Problems of Batch Processing of PIVAS in Our Hospital
Shan JIANG ; Ruoqiu FU ; Junhui REN ; Laichun LU ; Desheng MENG
China Pharmacy 2005;0(13):-
OBJECTIVE:To provide reference for the promotion of work efficiency of PIVAS.METHODS:The theory of bat-ch processing of PIVAS in our hospital was interpreted and its effect and problems were analyzed.RESULTS & CONCLUSIONS:Batch processing principal of background decision and foreground adjustment were adopted to allocate workload of different periods scientifically and improve working efficiency.Some problems existed in processing period require improvement of batch processing.
3.Safety of a novel biodegradable stent implanted in the coronary artery in a porcine model
Hu LI ; Xiaoyan LI ; Xuejun JIANG ; Xiaoxin ZHENG ; Gaoke FENG ; Suyuan HE ; Shan REN ; Xin YI
Chinese Journal of Tissue Engineering Research 2013;(38):6773-6778
BACKGROUND:At present, the main research fields about coronary stents are the whole degradation biological materials with high biocompatibility and drug control ed release systems.
OBJECTIVE: To evaluate the safety after the two novel biodegradable stents implanted in coronary arteries in porcine models.
METHODS:The normal ful y biodegradable stents were made up of the poly-L-lactide and the antiproliferative drugs paclitaxel, and the novel biodegradable stents were added in amorphic calcium phosphate at the basis of normal biodegradable stents. (1) Five normal ful y biodegradable stents were randomly implanted into the coronary arteries of five porcines, and five novel biodegradable stents were randomly implanted into the coronary arteries of the remaining five porcines by coronary angiography. The blood biochemistry and C-reactive protein levels were measured pre-operation and at 28 days after operation. Coronary angiography was utilized to observe the lumen unobstructed at 28 days after surgery. (2) Under a microscope, seven normal ful y biodegradable stents and seven novel biodegradable stents were implanted into right external iliac arteries of 14 rabbits. Blood urea nitrogen and creatinine levels were measured before surgery and at 28 days after operation.
RESULTS AND CONCLUSION:At 28 days after operation, there were no significant changes in porcine glutamic-pyruvic transaminase, aspartate aminotransferase, triacylglycerol, total cholesterol, low density lipoprotein and C-reactive protein levels compared with that before operation, but urea nitrogen and creatinine levels were significantly higher than that before operation (P<0.05). The result of coronary angiography showed that no in-stent thrombosis or stenosis was detected in either group. There was no significant difference in urea nitrogen and creatinine levels in both groups. These results suggested that it is safe and compatible after the two novel biodegradable stents implanted in coronary arteries of porcine models, and the stents had good histocompatibility.
4.MTT colorimetric assay of corneal epithelial cell activity
Huanwen, QIAN ; Jie, ZHANG ; Qing, SHAN ; Hua, REN ; Yanming, JIANG ; Denglong WANG
Bulletin of The Academy of Military Medical Sciences 2001;25(2):124-125
Objective:To establish a method for measuring the activity of cornea epithelium quantitatively. Methods:Rabbit corneas were burnt either by alkali or by CO2 laser. The lamellar cornea was cut at the end of 1,2 and 3 weeks and cultured in 2 ml DMEM with 5% CO2, 37℃ for 1 h.Then 200 μl of MTT was added to the culture followed by incubation for another 4 h. The supernatant was discarded and 4 ml of DMSO was added into each culturedish for dissolving MTT completely under the condition of room temperature.200 μl of DMSO sample was added to each well of 96-well plate and each sample was triplicated. The absorbance of the plate was measured at 490 nm ultraviolet. Results:The D value of the burnt corneas was obviously lower than that of the normal ones(P<0.01). Conclusion:MTT method can be used to measure the activity of cornea epithelium quantitatively.
5.Clinical application of conventional magnetic resonance imaging and diffusion-weighted imaging in differentiating histopathological types of small hepatocellular carcinoma
Jingjing LIU ; Jin WANG ; Ronghua YAN ; Bing HU ; Bingjun HE ; Zaibo JIANG ; Bihong LIAO ; Yingying LIANG ; Linglan REN ; Hong SHAN
Chinese Journal of Hepatobiliary Surgery 2012;18(8):573-577
Objective To study the imaging apperances and the diagnostic value of conventional magnetic resonance imaging (MRI) and diffusion-weighted imaging (DWI) in differentiating histopathological types of small hepatocellular carcinoma (sHCC).Methods 40 sHCC confirmed by histopathology were classified into 4 groups according to their degree of differentiation:well (n=6),well-moderate (n=5),moderate (n=27) and moderate-poor (n =2).All patients received conventional MRI and DWI (1.5T,b =0 and 600 s/mm2) before the operation.The ADC values of the sHCC were measured and compared.Results On T1WI,32 lesions showed hypointensity,4 hyperintensity (well) and 4 isointensity (well-moderate =2,moderate =2).On T2WI,hyperintensity was observed in 39 lesions and isointensity in 1 lesion (well).Steatosis in the sHCC was seen in 17 of 40lesions (17/40,42.5 %,well=4,well-moderate=1 and moderate=12).A pseudocapsule was seen in 67.5 % sHCC (27/40,well=4,well-moderate=3,moderate=18 and moderate-poor=2).32 lesions showed hypervascularity on arterial phase,and 8 lesions showed hypovascularity (well=3,moderate =4,moderate-poor=1).On DWI,37 lesions showed hyperintensity,except for 3 lesions with welldifferentiated sHCC which showed isointensity (50%,3/6).The mean ADC values±S.D.of sHCC in the well,well-moderate,moderate and moderate-poor groups were (1.757 ± 0.337) × 10-3,(1.917±0.574)×103,(1.816±0.545)×103 and (1.723±0.217)×10-3,respectively.There were no significant differences among the 4 groups.Conclusion The imaging appearances of wellmoderate,moderate and moderate-poor sHCC on conventional MRI were classical which make diagnosis easy.Hyperintensity on DWI contributed to diagnosis.However,the imaging appearances of some well-differentiated sHCC were atypical.The lesions could be isointensity or hyperintensity on DWI.The combination of conventional MRI and DWI contributed to better diagnosis of sHCC,especial for atypical sHCC.
6.Protective effects of mixture for protecting liver and supplementing stomach (保肝益胃合剂) on rat acute liver damage induced by carbon tetrachloride
Shu-Min JIANG ; Jing-Xi SHAN ; Jiang REN ; Bing LIU ; Bao-Zhu WANG ; Ke-Feng SUN ; Yong-Qiang FU ; Hui WANG ; Ying WANG ; Chun-lai YOU
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2006;0(04):-
Objective To observe the protective effects of a mixture for protecting liver and supplementing stomach (保肝益胃合剂) on rat acute liver damage induced by carbon tetrachloride (CCl_4). Methods The model of rat acute liver damage was established by injection of CCl_4 2 ml/kg into the abdominal cavity.The rat models were treated respectively by the mixture for protecting liver and supplementing stomach 30 g?kg~(-1)?d~(-1),the polyene phosphatide acid radical choline capsule [Yi Shanfu (易善复), 180 mg?kg~(-1)?d~(-1)],the glycyrrhizic acid diaminogen capsule [Gan Lixin (甘利欣),30 mg?kg~(-1)?d~(-1)] infused into the stomach.The activities of serum alanine transaminase (ALT) and aspartate transaminase (AST) were detected.In the mean time,the liver pathological changes were observed,the degree of liver cell necrosis was evaluated,and the rat mortality was noted in various groups of treatment.Results The values of ALT,AST and the score of liver cell necrosis in the group treated with the mixture for protecting liver and supplementing stomach [(1.168?1.066) kU/L,(1.845?2.212) kU/L,(0.56?0.53) score] were significantly lower than those in the model group [(4.982?3.502) kU/L,(7.030?3.616) kU/L, (1.38?0.92) scores],and all the differences being statistically significant (all P
7.Fusion expression of human renal cell carcinoma-associated antigen G250/MN/CA IX in prokaryotic expression system.
Yao-dong JIANG ; Shao-bin ZHENG ; Wang-long TAN ; Shan-chao ZHAO ; Fei REN ; Bao ZHANG
Journal of Southern Medical University 2007;27(3):307-309
OBJECTIVETo achieve high expression of human renal cell carcinoma-associated antigen G250 in Escherichia coli.
METHODSThe gene fragments encoding the protein obtained by PCR was cloned into prokaryotic expression vector pET32a(+) and expressed in E. coli Rosseta. The immunogenicity of the recombinant protein was evaluated by Western blotting.
RESULTSThe plasmid pET32a(+)/G250 was constructed and expressed in E. coli Rosseta successfully. Western blot analysis showed that the recombinant protein could be specifically recognized by monoclonal antibody M75.
CONCLUSIONEfficient G250 expression is achieved in prokaryotic expression system, which may facilitate further functional study of the protein and its monoclonal antibody preparation.
Antibodies, Monoclonal ; immunology ; Antibody Specificity ; immunology ; Antigens, Neoplasm ; genetics ; immunology ; metabolism ; Biomarkers, Tumor ; genetics ; immunology ; metabolism ; Blotting, Western ; Carbonic Anhydrase IX ; Carbonic Anhydrases ; genetics ; immunology ; metabolism ; Cloning, Molecular ; Escherichia coli ; genetics ; Gene Expression ; Humans ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism
8.Small interfering RNA targeting C-erbB-2 gene increases the radiosensitivity of lung adenocarcinoma cells in vitro.
Hua-Shan SHI ; De-Lian REN ; Xin-Mei CAO ; Li HUANG ; Yao JIANG
Journal of Southern Medical University 2008;28(11):1977-1980
OBJECTIVETo investigate the effect of small interfering RNA (siRNA) targeting C-erbb-2 oncogene on the radiosensitivity of C-erbb-2-overexpressing lung adenocarcinoma cell line.
METHODSFour pairs of siRNA targeting the coding sequence of C-erbb-2 mRNA were synthesized and their interference effects were evaluated using quantitative real-time fluorescence PCR. The siRNA with the best interference effects was transfected into Calu-3 cells, which were then exposed to 2 or 5 Gy irradiation, with the cells with transfection or irradiation alone as the control groups. The cell apoptosis after the treatment was detected using annexin V-FITC Kit.
RESULTSThe apoptosis rate of the Calu cells was 7.767-/+0.551 in the blank group, 14.400-/+1.114 in the interference group, 11.867-/+0.737 in 2 Gy irradition group, 23.000-/+1.664 in 2 Gy irradiation + interference group, 16.100-/+0.624 in 5 Gy irradiation group, and 27.900-/+1.709 in 5 Gy irradiation+interference group.
CONCLUSIONThe siRNA targeting C-erbb-2 gene can enhance the radiosensitivity of Calu-3 cells to gamma-ray and increase their apoptosis rate following gamma-ray exposure in vitro.
Adenocarcinoma ; genetics ; pathology ; Apoptosis ; drug effects ; radiation effects ; Cell Line, Tumor ; Genes, erbB-2 ; genetics ; Humans ; Lung Neoplasms ; genetics ; pathology ; RNA Interference ; RNA, Small Interfering ; genetics ; Radiation Dosage ; Radiation Tolerance ; genetics
9.Hypertonic stimulation induces synthesis and release of glutamate in cultured rat hypothalamic astrocytes and C6 cells.
Rong CAO ; Shan JIANG ; Li DUAN ; Ying-Fei XIONG ; Bei GAO ; Zhi-Ren RAO
Neuroscience Bulletin 2008;24(6):359-366
OBJECTIVETo investigate whether hypertonic saline (HS) can induce the synthesis and release of glutamate in cultured hypothalamic astrocytes or C6 cell line.
METHODSAstrocytes were isolated, cultured, purified and identified from the hypothalamus of newborn rat (1 day). The astrocytes were randomly divided into five groups: isotonic (IS) and HS groups, astrocytes were incubated by IS and HS (320 mosM NaCl) medium, respectively, for 1, 3, 5, 10 or 15 min; carbenoxolone (CBX)+IS and CBX+HS groups, astrocytes were pre-treated with CBX (100 mmol/L) for 1 h at 37 degrees C in a 5% CO(2) / 95% atmosphere, then removed to IS and HS medium, respectively, for 1, 3, 5, 10 or 15 min; Ca(2+)+HS group, astrocytes were pre-incubated with Ca Ca(2+) (1,000 micromol/L) for 1 h at 37 degrees C in a 5% CO(2) / 95% atmosphere, followed by a wash with isotonic FBS/DMEM, and then removed to hypertonic saline for 1, 3, 5, 10 or 15 min. The media of five groups were collected to analyze the medium glutamate concentration with high performance liquid chromatography. The astrocytes were fixed and double immunofluorescent stained with anti-glial fibrillary acidic protein (GFAP) and anti-glutamate. The C6 cells were divided into four groups: IS, HS, CBX+IS and CBX+HS groups, and used for quantitative measurement of glutamate in cells by flow cytometry (FCM).
RESULTS(1) Anti-GFAP immunofluorescent signal revealed no significant difference among various time points in each group, or among the five groups. (2) The anti-glutamate immunofluorescent signal was increased in HS group and peaked at 5 min, and decreased and returned to the level of IS group at 15 min (P < 0.01 vs the 5 min of HS group). In CBX+HS group, the glutamate intensity was higher than that in CBX+IS and HS groups. (3) The medium glutamate concentration had no change after treatment with HS for 1 and 3 min, while increased markedly after treatment for 5 min to 15 min (P< 0.01 vs 1 min and 3 min). On the contrary, the medium glutamate concentrations in the CBX+HS or Ca(2+)+HS group were significant lower than that in the HS group (P < 0.01). (4) FCM showed HS and CBX+HS induced glutamate increase in C6 cells.
CONCLUSIONHS induced cultured rat hypothalamic astrocytes or C6 cells to synthesize and release glutamate; CBX could block glutamate release, but could not disrupt glutamate synthesis.
Analysis of Variance ; Animals ; Animals, Newborn ; Astrocytes ; drug effects ; metabolism ; Calcium ; pharmacology ; Carbenoxolone ; pharmacology ; Cells, Cultured ; Chromatography, High Pressure Liquid ; methods ; Flow Cytometry ; Gene Expression Regulation ; drug effects ; Glial Fibrillary Acidic Protein ; metabolism ; Glutamic Acid ; metabolism ; Hypothalamus ; cytology ; Rats ; Saline Solution, Hypertonic ; pharmacology ; Time Factors
10.Enhanced nutritional therapy may promote wound healing after endoscopic therapy in patients with liver cirrhosis and esophageal varices.
Chang-zheng LI ; Qing-shan LI ; Xue LI ; Jun-hong YAN ; Rui-ling WANG ; Ren-xiu JIANG
Chinese Journal of Hepatology 2013;21(10):739-742
OBJECTIVETo investigate the effect of enhanced nutritional therapy on wound healing after endoscopic therapy in patients with liver cirrhosis and esophageal varices.
METHODSFifty patients with liver cirrhosis and esophageal varices were randomly divided into an enhanced nutritional therapy group (n = 25) and a control group (n = 25). The enhanced nutritional therapy group received one week of enhanced nutritional supplementation, including liver nutritional elements, prior to routine endoscopic therapy. The routine without any change to their diet. The rate of transformation and status of wound healing of esophageal varices were compared between the two groups.
RESULTSThe ratio of ulcers occurring at the injection site was lower in the enhanced nutrition group than in the control group (16/25 vs. 23/25; x2 = 5.711, P = 0.017). The enhanced nutrition group had only one case of minimal bleeding occurring during endoscopy as compared to the seven cases of bleeding in the control group (x2 = 5.357, P = 0.021). On average, the enhanced nutrition group required less sessions of endoscopic treatment to achieve eradication of esophageal varices than the control group (3.8 vs. 4.1; t = 2.069, P = 0.044).
CONCLUSIONPre-endoscopic enhanced nutritional therapy may benefit patients with liver cirrhosis and esophageal varices by promoting recovery of procedure-related local tissue injury and occlusion of varices.
Adult ; Endoscopy ; Esophageal and Gastric Varices ; etiology ; therapy ; Female ; Humans ; Liver Cirrhosis ; complications ; therapy ; Male ; Middle Aged ; Nutritional Support ; Wound Healing