Animals ; Cells, Cultured ; Female ; Granulosa Cells ; metabolism ; Pituitary Adenylate Cyclase-Activating Polypeptide ; genetics ; metabolism ; Platelet Activating Factor ; pharmacology ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley

Abdominal Cavity ; blood supply ; Adolescent ; Angioplasty, Balloon ; Child ; Female ; Humans ; Male ; Takayasu Arteritis ; therapy

BACKGROUND: Some researches suggest that placenta is regarded as a new source of mesemchymal stem cells (MSCs). Adherent cells derived from placenta tissue have similar morphological characteristics and surface markers to MSCs; meanwhile, they can differentiate into osteoblasts and nerve cells.OBJECTIVE: To find a new source and way for MSCs separation.DESIGN: Observational study.SETTING: The Third People's Hospital of Wuxi.MATERIALS: Placenta was sourced from cesarean in our department of obstetrics and gynecology and provided confirmed consent from the relatives and ethics committee. The main reagents contained culture medium, sABC kit, DAB staining kit, caprine-anti-rat FITC, recombinant human basic fibroblast growth factor (rh-bFGF), rabbit-anti-human glial fibriliary acidic protein (GFAP).METHODS: The experiment was carried out in the Laboratory of Cells & Molecular Biology, the Third People's Hospital of Wuxi from May 2005 to August 2006. Placenta entity tissue was digested, adherent cells were cultured, morphological characteristics were observed, and growth curves of placenta-derived multipotent cells (PDMCs) in various generations were drawn. On the 1st and 7th days, supernatant was derived from cells in primary culture to measure content of β-glycerophosphate disodium (β-HCG) with chemiluminescence technique. In addition, expression of surface antigen and differentiating potency were detected at the same time. At 24 hours (at phase of nerve cells) and 2 weeks (at phase of osteoblasts) after inducible differentiation, cultured cells were dealt with routine immunocytochemical stain, and then they were observed under routine microscope or fluorescence microscope.MAIN OUTCOME MEASURES: ① Morphological indexes and growth curves of PDMCs; ② measurement of β-HCG in supernatant of PDMCs and expression of antigen of PDMCs with flow cytometer; ③ immunohistochemical analysis of PDMCs during and after inducible differentiation.RESULTS: ① Primary culture of PDMCs: After digestion of placenta tissue, a few of adherent cells were obtained and gradually formed thin and flat monolayer cells two weeks later. The monolayer cells grew like whirlpool or cluster. With the increase of cell density, soma was slenderer and slenderer and like fibroblast. ② Growth curves of PDMCs: Growth latency ranged from 2 to 8 days after cell inoculation. During this period, adherence was observed gradually but not obviously amplified. Eight days later, cells entered log growth phase. During this period, proliferation was active and cell process surrounding extended under phase contrast microscope. A lot of MSCs were observed in division phase between two nuclei. In addition, density was increased and cells connected to each other. Within 11-14 days, growth curve gradually entered platform phase. MSCs covered the bottom of bottle, cells slowly expanded, and primary culture stopped. ③ Measurement of β-HCG: Expression of β-HCG was not detected in supernatant at two time points. ④ Characteristics of surface antigen of PDMCs: PDMCs could express CD29, CD44 and CD105, but not CD34, CD45,CD19 and CD106. ⑤ Inducible differentiation of PDMCs: At 24 hours after inducing to nerve cells, form of PDMCs obviously changed, soma rebounded, refraction of nucleus was partially reinforced, structure was similar to dendrite and axis-cylinder, and positive neuro-specific enolase and GFAP were observed after staining.CONCLUSION:Placenta tissue contains PDMCs whose morphological function is similar to MSCs; in addition, placenta is regarded as an effective source of MSCs.

BACKGROUND: Multipotential mesenchymal stem cells (MSCs) can be differentiated into bone, fat, cartilage, muscle and endothelial cell at the specific conditions, so it draws great attentions of the related study.OBJ ECTTVE: To establish the method of in vitro culture and expansion of human umbilical blood-derived MSCs and investigate their influencing factors.DESTGN: Randomized and controlled trials.SETTING: Department of Tissue Engineering and Cell Biology in Wuxi Third People's Hospital.MATERTALS: Seventy cases of human umbilical cord blood (HUCB) of healthy neonates (selected from Parturient Room, Department of Gynecology and Obstetrics, Wuxi Third People's Hospital, with the informed consents of the puerpera and their relatives), 40 mL each; dulbecco's minimal essential medium (DMEM) of low glucose (LG) at the dose of 50 g/L, fetal bovine serum (FBS), mycillin and trypsin (all were from Gibco), FITC-labeled mice anti-human CD29, CD105 and CD106 (Ancell) monoclonal fluorescent antibody, PE-labeled mice anti-human CD34, CD44, CD45, CD19, HLA-DR (Immunotech)monoclonal fluorescent antibody, Ficoll separation medium (Pharmacia).METHODS: The experiment was carried out in Department of Tissue Engineering and Cell Biology in Wuxi Third neonatal cord blood were anticoagulated by 25 u/mL heparin, and 60 cases of isolated HUCB mononuclear cell were precipitated and suspended by cell culture fluid (50 g/L DMEM-LG + 50 g/L FBS + 10 g/L mycillin), while other 10 cases were purified with DMEM of high glucose (100 g/L) to produce adherent layer. Once the cells reached fluences of FBS coating on adhesive rats of MSCs: Some of the purified HUCB MSCs at logarithmic phase were growth curve of MSCs: The growth of cells were observed by phase contrast microscope (OLYMPUS CK40), and were detected by EPICS-ALTRA flow cytometry.FBS coating and without FBS coating at the different time points, and their adhesive rates.RESULTS: All 70 cases of HUCB MSCs were adopted in this study, however, 10 samples failed to obtain the ideal MSCs cord blood MSCs after culture. During 2 weeks of culture, primary cells began to double for 3-4 days and reached conand CD105, but did not express antigens CD34, CD45, CD106 and HLA-DR, which was identical to surface labeling of those without FBS coating (P＜0.01).CONCLUSrON: MSCs in HUCB can be cultured and expanded in vitro, and are regarded as an alternative source of MSCs for experimental and clinical applications. Moreover, high glucose can depress the growth and proliferation of MSCs in HUCB.

Objective To investigate the effect of cholecystokinin (CCK) on colon motility and its mechanism in development of irritable bowel syndrome via recording ionic channels currents and contraction of guinea-pig proximal colon. Methods The guinea-pigs (body weight ranged from 200 g to 250 g) were deprived of food, but not water, for 12 hours before experiment. The animal was sacrificed and 6 cm of proximal colon was obtained. The contractile activity of isolated proximal colon in 1 × 10-7 ,5 × 10-7 or 1 × 10-6 mol/L of CCK-8 solution was recorded. The impact of 1 × 10-7 , 5 × 10-7 and 1 × 10-6 mol/L of CCK-8 and 1 × 10-6 mol/L CCK-8 nifidipin on current of calcium activated potassium channel (IBKac) was detected with an EPC-10 amplifier and an image analysis software.Results In comparison with blank [(0. 68 ±0. 12) g], the amplitude of colon contraction in 1 × 10-7 ,5×10-7 and 1×10-6 mol/L of CCK-8 was increased by (15. 0±1.5)%,(28. 0±1.4)%, and (36.0±1.6) %, respectively ( n = 7, P = 0. 023,0. 005 and 0. 01 ), but there was no significant change of frequency. When exogenous stimulation at +60 mV, the current of IBKac was enhanced to (117. 45 ± 3.60)%, (125.42± 5. 30)% or (136. 98±6. 80)% in 10-7 ,5 × 10-7 or 10-6 mol/L of CCK-8,respectively, compared with controls (n= 7, P＜0.01 ). However, after adding nifidipin, the current of IBKca was reduced to (102.23±5.60)% in 10-6mol/L of CCK-8 at +60 mV when compared with controls (n=7, P= 1. 491 ). Conclusion CCK enhances proximal colonic motility by increasing Ca2+ influx and IBKac current, which is characterized by enhancement of amplitude of contraction.

The effect of RNA interference-mediated adiponectin gene silence on glucose-lipid metabolism in apolipoprotein E gene knock-out(ApoE~(-/-)) mice was studied. The plasma adiponectin level in ApoE~(-/-) mice with high-fat diet and adiponectin short hairpin RNA (shRNA) adenovirus injection (ADI group) was significantly lower than that in ApoE~(-/-)mice with high-fat diet(HF group), normal-chow(NF group)and naked adenovirus injection (GF, both P<0.01). Fasting blood glucose, fasting plasma insulin, free fatty acids, total cholesterol, triglycerides, low-density lipoprotein-cholesterol (LDL-C), high-density lipoprotein-cholesterol (HDL-C), and body weight were significandy elevated in three high-fat diet groups compared with in NF group, (P<0.01). However, fasting plasma insulin, free fatty acids, total cholesterol, triglycerides, LDL-C, and HDL-C in ADI group were significantly higher than those in HF and GF groups(P<0.01 or P<0. 05). A typical clinical phenotype of glucose-lipid metabolism and insulin resistance in ApoE~(-/-) mice can be induced by high-fat feeding and adiponecting gene silencing.

Objective To study the thrombolysis effect and safety of UK combined with tirofiban in young patients with ST segement elevation acute myocardial infarction.Methods 76 young patients with ST segement elva-tion acute myocardial infarction were selected as research objects,and they were divided into UK group(control group, n =34)and UK combined with tirofiban group(observation group,n =42)by random number table,then the recanali-zation rate of infarct -related artery and incidence of complications were compared.Results The recanalization rate of infarct -related artery in the observation group was 97.6%,which was 91.2% in the control group,there was no significant difference between the two groups(χ2 =1.564,P >0.05);The vascular infarction related to blood flowⅡlevel of the patients was 90.5% in the observation group,which was significantly higher than 70.6% in the control group(χ2 =4.945,P <0.05);The death,complications incidences had not significant differences between the two groups(χ2 =1.252,2.837,2.837,all P >0.05).Conclusion The thrombolysis effect of UK has a high recanaliza-tion rate of infarct -related artery in young patients with ST segement elevation acute myocardial infarction,and the thrombolysis effect of UK combined with tirofiban is better,and it does not increase the incidence of complications.

Objective:To study dynamic changes of fragile histidine triad(FHIT) gene expression in ionizing radiation injury and radiation carcinogenesis. Methods: BEAS-2B cells were divided into 0.5,1,2,4,8,16 Gy irradiation groups and control group. In 24 h, 72 h and 10 d after irradiation, the expression of FHIT gene was studied with single-cell RT-PCR and DNA sequencing separately. Results: Different types of FHIT gene mutations occured in different phases after irradiation with different doses (All mutations were exon deletion mutations by DNA sequencing), while abnormal FHIT gene was not detected in control group. The percentage of mutation in 0.5,1,2,4,8,16 Gy dose groups was 52.6%,66.7%,57.9%,76.5%,64.7% and 81.3% respectively 24 h after irradiation;17.6%,22.2%,50.0%,47.4%,47.1% and 68.4% respectively 72 h after irradiation;and 21.1%,25.0%,60.0%,57.9%,61.1% and 68.4% respectively 10 d after irradiation. Conclusion: These results suggest that ionizing radiation can cause deletion of FHIT gene.

Objective Establish a laboratory quality control system to ensure accurate and reliable test data and to contrapose the influence of error factors in current detection methods for urinary iodine measurement. Methods The results of reagent blank absorbance value and uric iodine standard materials were collected, then their relevant technical indexes such as mean, standard deviation, control limit, auxiliary line were worked out. Then the quality control chart of blank test and the relative error control chart were made base on these technical indexes. And different iodine concentrations (high, middle and low concentration) were tested and their mean,relative reduction difference value, weighted mean value and critical limit Rc value were calculated, and then critical limit Rc value precision control chart was made. Results The range of absorbance of blank control test was 1.183 to 1.553. And the limit of the accuracy control Rc value was 0.0883, 0.0572, respectively, when the concentrations of urinary iodine was 0～ < 150 μg/L and 150 ～ 300 μg/L. The control limit of the relative error was 9.3%. Conclusions The method of quality control chart could be satisfactorily applied to identify the quality of the analytical results of urine iodine, and ensure the results of the urine iodine reliable and authentic.

Objective To analyze the antibiotic resistance of Branhemella catarrhalis strains isolated from sputum specimens of patients with lower respiratory tract infections from Linyi, Shandong Province, and to explore the relationship between bro genotypes of the strains and their resistance to antibiotic agents.Methods Sputum specimens were colleted from the patients with lower respiratory tract infections in Linyi People ’ s Hospital from the January 2010 to December 2014.The specimens were inoculated into 4 different disks for bacterial isolation and cultivation.β-lactamase detection and drug sensitivity tests were performed, and PCR coupled with restriction endonuclease analysis was employed for bro genotyping.χ2 test was used to compare drug resistance of strains with different bro genotypes.Results A total of 497 Branhemella catarrhalis strains were isolated in five years, among which 221 strains were isolated in winter.All strains were sensitive to ertapenem and chloramphenicol, and the resistance rates to amoxicillin/clavulanate and cefaclor were low (≤2.8%).The strains were highly resistant to compound sulfamethoxazole, erythromycin and ampicillin (47.6%-89.8%), and there was a trend of increasing resistance rates with the year, but no statistically significant difference was observed ( P >0.05 ) .β-lactamases was positive in 412 strains (82.9%), and all of these strains were positive for bro gene, and the resistances to erythromycin, compound sulfamethoxazole, levofloxacin and ampicillin were higher in bro positive strains than those in bro negative strains (χ2 =12.16, 16.18, 8.41 and 200.00,P<0.05).Among bro positive strains, 391 (94.9%) were of genotype bro-1, 21 (5.1%) were of genotype bro-2, and their resistance to antibiotic agents was not of statistical difference ( P >0.05 ).Conclusions Most of Branhemella catarrhalis clinical isolates are β-lactamase producing strains, and bro-1 is the most common genotype.Strains are highly sensitive to carbapenems, cephalosporins andβ-Lactamaseinhibitors, which can be recommended for the treatment of Branhemella catarrhalis-related respiratory tract infections.