BACKGROUND: Recent studies indicate that after indirect co-culture of neonate rat myocardial cells and bone marrow mesenchymal stem cells, bone marrow mesenchymal stem cells can differentiate into myocardial cells successfully. OBJECTIVE: To induce human bone marrow mesenchymal stem cells (hBMSCs) to differentiate into endothelial cells using human umbilical vein endothelial cells by indirect co-culture. DESIGN, TIME AND SETTING: In vitro study of cell engineering was done at the Medical Research Center of Guangdong Provincial People’s Hospital between January and July 2007. MATERIALS: Small quantities of bone marrow were obtained from 11 children with congenital heart disease but without hematologic diseases through manubrium of sternum puncture in the congenital heart defect corrective surgery after the permission of family member. Umbilical cord of full-term normal delivery healthy newborn was provided by the Department of Obstetrics, the First Affiliated Hospital of Sun Yat-sen University. Fresh cattle jugular vein was provided by Guangdong Dali Meat Cattle Butchery. METHODS: The hBMSCs were isolated and purified using density gradient centrifugation method and were cultured in vitro. Human umbilical vein endothelial cells were obtained from newborn umbilical cord by enzyme digestion. Cell culture insert with semipermeable membrane combined with 6-well plate was used to do indirect co-culture induction. Human umbilical vein endothelial cells were expanded in the cell culture insert, passage 3 bone marrow mesenchymal stem cells were expanded in the 6-well plate outside the culture insert at the density of 1?105 cells/well, the initial ratio of the two kinds of cells was 1:5, then low-glucose DMEM culture solution containing 10% fetal bovine serum was added, cells were cultured for 14 days. Co-culture of bone marrow mesenchymal stem cells and bone marrow mesenchymal stem cells was used as control. Introduced endothelioid cells were cultured and then seeded on the cell-free cattle jugular vein intravascular stent. MAIN OUTCOME MEASURES: Morphology changes of induced cells; introduced endothelioid cell surface antigen detected through immunocytochemical staining; the growth and adhesion condition of endothelioid cells on the intravascular stent observed under scanning electronic microsope. RESULTS: The morphologies of introduced endothelial cells were uniform, introduced endothelial cells presented a cobblestone-like appearance, they amplified fast and expressed endothelial cell-specific surface marker CD31 and vWF and the positive rate was over 99%. They also could form a continuous unicellular layer on the cell-free cattle jugular vein intravascular stent. CONCLUSION: Human umbilical vein endothelial cells can induce hBMSCs to differentiate into endothelial cells successfully and to adhere and grew on the cell-free cattle jugular vein intravascular stent through indirect co-culture method.

Objective To analyze the prevention and management of in-hospital complications to the surgical treatment of hip fractures in elder patients (≥80 years). Methods An retrospective data of 198 hip fracture patients with surgical treatment between Jan. 1996 and Mar. 2004 was reviewed. Their age ranged from 80 to 98 years with an average of 85.2 years. 93 cases were femoral neck fracture (Garden type Ⅰ 2 cases, type Ⅱ 10 cases, type Ⅲ 52 cases and type Ⅳ 29 cases) while 105 were intertrochanteric fracture ( Evan type Ⅰ 3 cases, typeⅡ 37 cases, type Ⅲ 39 cases and type Ⅳ 26 cases). Local anaesthesia, general anaesthesia and epidural anaesthesia were used in 2, 38 and 158 cases respectively. Femoral neck fractures were treated with bipolar femoral head replacement in 77 patients, total hip replacement in 13 patients, and compressive cannulated screw fixation in 3 patients respectively. Intertrochanteric fractures were treated with close reduction and dynamic hip screw (DHS) internal fixation. Results The average operation duration and blood loss were 53 min and 110 ml respectively in the bipolar femoral head replacement group, 94 min and 165 ml in the total hip replacement group, 35 min and 30 ml in the screw fixation group and 40 min and 60 ml in the DHS group. 17 cases under cemented hip replacement suffered obviously transient arterial blood pressure drop. 36 cases (18.2%) had postoperative complications, among which the mental system and circulation system were more inclined to be involved. The average in-hospital time was 18.6 days and 2 cases died in hospital. The average time from admission to operation (waiting time ) was 6 days. Among 46 cases longer than 7 days, 9 cases (19.6%) had postoperative complications. Among 152 cases no longer than 7 days, 27 cases (17.8%) had postoperative complications. It showed no statistical significant difference between the two groups in complication incidence. Conclusion To gain good results in elderly patients of hip fracture by operation, less invasive and quick anaesthetic technique and operative procedures are encouraged. Meticulous preoperative preparation, active preventing and treating the intraoperative and postoperative complications should be emphasized on.

BACKGROUND:Retinoid X receptor (RXR) plays a central role in the regulation of intracellular receptor signaling pathways. The activation of RXR has protective effect on H2O2-induced apoptosis of H9c2 ventricular cells in rats. But the protective effect and mechanism of activating RXR in cardiomyocytes against hypoxia/reoxygenation (H/R)-induced oxidative iniury are stillunclear. METHODS:The model of H/R injury was established through hypoxia for 2 hours and reoxygenation for 4 hours in H9c2 cardiomyocytes of rats. 9-cis-retinoic acid (9-cis RA) was obtained as an RXR agonist, and HX531 as an RXR antagonist. Cultured cardiomyocytes were randomly divided into four groups:sham group, H/R group, H/R+9-cis RA -pretreated group (100 nmol/L 9-cis RA), and H/R+9-cis RA+HX531-pretreated group (2.5 μmol/L HX531). The cellviability was measured by MTT, apoptosis rate of cardiomyocytes by flow cytometry analysis, and mitochondrial membrane potential (ΔΨm) by JC-1 fluorescent probe, and protein expressions of Bcl-2, Bax and cleaved caspase-9 with Western blotting. Allmeasurement data were expressed as mean±standard deviation, and analyzed using one-way ANOVA and the Dunnett test. Differences were considered significant whenP was <0.05. RESULTS:Pretreatment with RXR agonist enhanced cellviability, reduced apoptosis ratio, and stabled ΔΨm. Dot blotting experiments showed that under H/R stress conditions, Bcl-2 protein level decreased, while Bax and cleaved caspase-9 were increased. 9-cis RA administration before H/R stress prevented these effects, but the protective effects of activating RXR on cardiomyocytes against H/R induced oxidative injury were abolished when pretreated with RXR pan-antagonist HX531. CONCLUSION:The activation of RXR has protective effects against H/R injury in H9c2 cardiomyocytes of rats through attenuating signaling pathway of mitochondria apoptosis.

Adult ; Device Removal ; methods ; Ductus Arteriosus, Patent ; Heart Septal Defects, Atrial ; therapy ; Humans ; Male

Fura—2 was used as a Ca~(2+) indicator to determine the intracellular calcium ion concentra-tion(〔Ca~(2+)〕i)of rat peritoneal macrophages(RPM?s),and APAAP enzyme immnoassay was ap-plied to detect the expression of Ia antigens on RPM?s.The results showed that norepinephrine(NE,10~(-9)mol/L)could markedly increase the〔Ca~(2+)〕i of the RPM?s(p

Objective:To investigate the regulation mechanisms of the bone marrow mesenchymal stem cells on lymphocyte pro -liferation of type I diabetic rats .Methods:The rat bone marrow mesenchymal stem cells were isolated , cultured and identified and the effect on lymphocyte proliferation of type Ⅰdiabetic rat was observed by MTT assay , and analyze the CD 4 +CD25 +regulatory T cell ra-tio, cell cycle and apoptosis of type I diabetes rat by flow cytometric .Results:B and C groups was significantly lower than the absor-bance values of group A,the differences between the data were statistically significant (P<0.05), C group was significantly lower than group B absorbance values, the difference was significant (P<0.05);the CD4 +CD25 +regulatory T cells of B and C groups were sig-nificantly higher than group A, the differences of the data were statistically significant (P<0.05), the CD4 +CD25 +regulatory T cell ratio of C group significantly higher than that group B , the differences were statistically significant (P<0.05);the apoptosis levels of B and C groups were significantly higher than group A , the differences were statistically significant (P<0.05), the apoptosis levels of C group were significantly higher in group B , the differences were statistically significant (P<0.05).Conclusion:Bone marrow mesen-chymal stem cells can significantly inhibit lymphocyte proliferation of type Ⅰdiabetic rats, and it may regulate CD4 +CD25 +regulatory T cells, promote apoptosis, thereby affecting the immune function of T lymphocytes , and play its rejection.

Objective To explore the expression and significance of hedgehog signal molecules (Ptch,Smo and Gli1 ) in chronic pancreatitis tissues in rats.MethodsSixty SD rats were randomly divided into CP group (n =50) and control group (n =10).DBTC solvent (8 mg · ml-1 · kg-1 ) was injected into the rat via tall vein in CP group.In control group,rats were treated only with the solvent at a dose of 1ml/kg body weight.All rats were sacrificed 6 weeks later to observe the pancreatic pathologic changes.Collagen accumulation in pancreatic sections was determined by staining for Sirius red.Expressions of Ptch,Smo,Gli1 mRNA and protein in pancreatic tissues were assessed by RT-PCR and immunohistochemistry.ResultsThe rate of chronic pancreatitis development in rats in CP group within six weeks was 73.9％.Collagen content was markedly higher in CP group than that in control group [ ( 38.52 ± 6.49 ) ％ ~s (7.37 ± 2.28 ) ％,P ＜ 0.05 ].No Path,Smo,Gli1 protein expression was observed in normal pancreatic tissues in control group.The positive rate of Ptch,Smo,Gli 1 expression was 73.5％,64.7％ and 52.9％ in CP group,and the difference between the two groups was statistically significant (P ＜ 0.05).The expressions of Ptch,Smo,Gli1 mRNA were 2.38 ±0.42,3.85 ± 1.03,4.63 ± 1.49 in CP group,which were significantly higher than those in control group (0.23 ±0.16,0.14 ±0.05,0.57 ±0.12,P ＜0.05).ConclusionsThe Ptch,Smo,Gli1 was highly expressed in pancreatic tissues in CP rats,suggests hedgehog messenger pathway may play an important role in the chronic inflammation and fibrosis of chronic pancreatitis.

AIM:To investigate the expressions of BAG-1 and NF-?B P65 in the multiple myeloma(MM)and approach the effects of BAG-1 and NF-?B P65 in the pathogenesis of MM and their clinical significance,which would be useful to find a theoretical basis for the treatment of MM.METHODS:The patients diagnosed MM in Yijishan hospital of wannan medical college from 2006.10 to 2008.10 as the experimental group,while the bone trauma patients selected as the control group.The mononuclearcells of bone marrow in the patients were extracted and the BAG-1and NF-?B P65 mRNA RT-PCR were detected.RESULTS:The expression of BAG-1 mRNA(71.4%)in the MM patients was higher than that in the control group(P=0.007).The expression of NF-?B P65 mRNA(75.0%)in the MM patients was higher than that in the control group(P=0.004).There was a positive correlation between the BAG-1 and NF-?B P65 mRNA.The tumor load was high in the MM patients with overexpression of BAG-1 and NF-?B P65,which was ralated to a number of clinical parameters indexes.BAG-1 and NF-?B P65 were positive expressed of MM tumor load,relatived with a number of clinical indicators.CONCLUSION:The expressions of BAG-1 and NF-?B P65 in MM are high,there are no difference between the experimental group and control group.BAG-1 and NF-?B P65 were related to a number of clinical indexes.BAG-1 and NF-?B P65 may be involved in the occurrence and the development of disease,and they can be used as one of the prognosis indexes.

The purpose of quality control is to form a standard,scientific,and unified control system for PET-CT.The improvement of regulations is the base.The training of technicians is the key.The establishment of radiological protection rules is the assurance.So it is an important link to improve the quality control of PET-CT for better sensitivity and accuracy of PET-CT.

Objective To assess the mortality,complications and major morbidity of pneumonectomy for non -small cell lung cancer(NSCLC)and to establish the importance of various prognostic factors.Methods The data of 64 consecutive patients who underwent pneumonectomy for NSCLC were retrospectively reviewed.Results The 30 -day mortality rate was 7.8%(5 /64).Complications developed in 29.7%(1 9 /64)and overall 5 -year survival was 1 9.0%.Pathological stage Ⅲ(P =0.030)and right pneumonectomy(P =0.01 0)were independent risk factors of an adverse outcome.Survival was not significantly influenced by histological types (P =0.1 25)or curability (P =0.587).Conclusion Pneumonectomy is associated with acceptable overall morbidity and mortality.However,the patients with pathological stage Ⅲ or right pneumonectomy require special consideration.Pneumonectomy should be performed only in selected patients.