BACKGROUND:Tissue and cellimplantation entails high-quality seed cells. In order to satisfy this requirement, it is crucial to produce adequate wel-conditioned, high-purity and strong proliferation ability bone marrow-derived mesenchymal stem cells.
OBJECTIVE:To establish a simple, rapid and effective in vitro isolation and culture method of bone marrow-derived mesenchymal stem cells, and to define the biological features of bone marrow mesenchymal stem cells.
METHODS:Rat bone marrow mesenchymal stem cells were isolated from the bilateral tibial and femoral bones by the method of whole bone marrow, then purified and passaged by attachment method. The morphology and features of bone marrow mesenchymal stem cells were observed, the growth curve was drawn and the cellsurface antigen was detected by flow cytometry. The bone marrow mesenchymal stem cells were induced to differentiate along the osteogenic, chondrogenic and adipogenic lineages.
RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cells isolated by the whole bone marrow adherence method grew vigorously and were highly purified. The cultured cells were spindle-shaped. The growth curve was S-shaped and the population doubling time was 29 hours. The cells stil maintained a strong proliferative capacity after they were passaged for 10 generations. The surface markers such as CD44, CD29, CD90 were positive, while CD45, CD34, CD11b were negative. At the third passage, bone marrow mesenchymal stem cells were induced to differentiate along the osteogenic, chondrogenic and adipogenic lineages, respectively. Fol owing induction, Alizarin red staining, alkaline phosphatase staining, von-kossa mineralized nodules staining, toluidine blue staining, and oil red O staining were al positive. This shows that the whole bone marrow adherence method is a simple and reliable method for the in vitro isolation, culture and proliferation of bone marrow mesenchymal stem cells. Moreover, they have multi-lineage differentiation capacity under different inducers. The third passage bone marrow mesenchymal stem cells have the highest biological activity and can act as the ideal seed cells for subsequent experiments.

Objective To analyze postoperative complications of surgical therapy for deep vein thrombosis (DVT) of the lower extremity. Method From January 2001 to January 2008 vena cava filters were placed in 171 DVT cases before surgery. Fogarty catheter (73 cases), Amplatz thrombectomy device (ATD) (55 cases), Acolysis ultrasound ablation(43 cases) were used to extract the thrombi in iliac and proximal femoral vein. The full extraction of thrombi in distal end was facilitated by compressing and massaging the legs in all cases. Iliac venous stenosis or occlusion was managed by interventional therapy, and temporary femoral arteriovenous fistula were carried out routinly. Result Operations were successful in 157 cases(70 cases in Fogarty group, 52 in ATD group,35 in Acolysis ultrasound ablation group), and failed in 14 cases (3 cases in Fogarty group, 3 in ATD group, 8 in Acolysis ultrasound ablation group). Permanent,retrievable and temporary vena cava filters were placed in 51,32 and 88 cases respectively. Stenosis or occulsions of the iliac vein was found in 143 cases and stents were placed after balloon dilatation in 41 cases. During the operation, residual thrombi was found in 80 cases. Vessel perforation and rupture were complicated in 14 cases, thrombosis adhering in eava vein filters in 18 cases. There was no mortality and no pulmonary thromboembolism. During the follow-up, we found iliae vein restenosis in 21 cases, thrombosis recurrence in 36 cases, stents displacement in 6 cases, and stents fracture in 2 cases. Conclusion Surgical therapy is effective for DVT of the lower extremities.

Background Diabetic retinopathy (DR) is a chronic inflammatory disease,with pathological changes of retinal microvessels.Studies demonstrated that sericin has anti-inflammation and anti-oxidation effects,inferring that sericin might play a protective effect on diabetic microangiopathy.Objective This study was to investigate the effects of sericin on intercellular adhesion molecule-1 (ICAM-1),vascular cell adhesion molecule-1 (VCAM-1) and Cx43 expressions in retina and explore the protection of sericin to retinal microangiopathy in diabetic rats.Methods Forty-eight specific pathogen free male SD rats were randomly divided into normal control group,diabetic model group,sericin-treated group and calcium dobesilate-treated group,with 12 rats for each group.The diabetic models were established by intraperitoneal injection of streptozotocin (STZ) for 3 consecutive days and feeding up with high lipid foods.Normal saline solution,2.4 g/(kg · d) sericin solution and 0.2 g/(kg · d) calcium dobesilate were used by gavage administration in the rats of the diabetic model group,sericin-treated group and calcium dobesilate-treated for 3 months group,respectively.The rats were sacrificed and retinal sections were prepared,and the retinal morphology was examined by hematoxylin-eosin staining.The expressions of ICAM-1,VCAM-1 and Cx43 proteins and mRNA in retinas were detected by Western blot assay and reverse transcription PCR.The use and care of the rats complied with Regulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission and ARVO statement.Results The retinal structure was normal in the normal control group.The swell and rupture of inter limiting membrane (ILM),scatter vascular endothelial cell nuclei breakthrough ILM and decrease of retinal ganglion cells (RGCs) were displayed in the diabetic model group;while in the sericin-treated group and calcium dobesilate-treated group,the mild thickening of ILM and disorder of retinal cells were obtained.The relative expression levels of ICAM-1 and VCAM-1 were significantly raised and those of Cx43 were reduced in the diabetic model group,sericin-treated group and calcium dobesilate-treated group when compared with the normal control group (all at P＜0.05).Compared with the diabetic model group,the expressions of ICAM-1 and VCAM-1 proteins and mRNA in the sericin-treated group were significantly reduced (ICAM-1 protein:0.834 3±0.032 1 vs.0.918 9±0.042 4;VCAM-Ⅰ protein:0.726 4±0.011 2 vs.1.235 0±0.078 9;ICAM-1 mRNA:0.716 3±0.008 6 vs.0.956 8±0.012 5;VCAM-1 mRNA:0.393 7±0.035 0 vs.0.477 9±0.020 6) and those of Cx43 protein and mRNA were evidently elevated (Cx43 protein:0.133 1 ±0.015 3 vs.0.039 2±0.002 0;Cx43 mRNA:0.676 8 ±0.064 8 vs.0.430 8±0.111 3) (all at P＜0.05).Conclusions Sericin can relieve retinal microangiopathy and protect retina against the pathogenesis and development of DR by down-regulating the expressions of ICAM-1,VCAM-1 and upregulating the expression of Cx43 in retinas of diabetic rats.

Objective To investigate changes of surface electromyogr8phic(sEM G)signal of pelvic floor muscles in complete spinal cord injury(cSCI)patients with neurogenic bowel dysfunction.Methods Fifteen hospitalized patients with cSCI(observation group)and fifteen normal subjects(control group)were involved in this study.The root mean square(RMS)of sEMG signals were collected at pelvic floor muscles with rectal surface electrode when subjects'pelvic floor muscles were rest(10 s),fleetly contract(2 s×3),continually contract(10 s).Both groups'data of different contracting states of pelvic floor muscles were analyzed and compared.Results The max RMS and average RMS(16.61±2.83,13.52±2.22)at pelvic floor muscles'rest in observation group were higher than that in control group(8.41±5.55,3.45±1.53).There was statistical difference between two groups(P<0.01).In the subjects of observation group max RMS and average RMS(20.24±13.99,13.36±2.39)at continual contraction and average RMS(13.40±2.31)at fleet contraction were nearly the same as RMS value at pelvic floor muscles'rest.There was no statistical difference between these two states(P>0.05).Conclusion The sEMG could be a quantitative index in assessing function of pelvic floor muscles and the neurogenic bowel dysfunction after cSCI.It can supply some clinical value in framing the training of pelvic floor muscles and improving the bowel dysfunction.

Objective To investigate the effect of HbA1c meeting the standard or not on microalbuminuria,blood lipids and liver enzymes in patients with type 2 diabetes.Methods A retrospective analysis was performed on 457 subjects who had type 2 diabetes.They were divided into substandard group and standard group according to HbA1c result.The general information and relevant laboratory indicators of patients were.collected and compared between two groups.Results The microalbuminuria,serum triglyceride and liver enzymes (glutamyl transpeptidase,alkaline phosphatase,aspertate aminotransferase) were significantly different between two groups [ (189.8 ± 235.3) mg/dl vs (38.9 ± 85.5) mg/dl,(2.64 ± 2.99) mmol/L vs (2.02 ± 1.50)mmol/L,(41.7 ±52.9)U/L vs (29.7 ±24.9)U/L,(83.6 ±28.6) U/L vs (74.3 ±25.8)U/L,(26.7 ±19.1)U/L vs (22.0 ±10.5) U/L,P ＜0.05].HbA1c level was positively correlated with microalbuminuria,glutamyl transpeptidase and alkaline phosphatase (r =0.209,0.115,0.11,P ＜0.01).The microalbuminuria was an independent risk factor of affecting HbA1c to reach the standard (OR = 1.009,P ＜0.05).Conclusions HbA1c meeting the standard or not can influence many factors except blood glucose.

Objective To evaluate the efficacy of endovascular interventional treatment for infrapopliteal arterial ischemic diseases. Methods Sixty patients(65 limbs)of infrapopliteal arterial isehernia of the lower extremities received pereutancotm transluminal angioplasty(PTA)and/or stents implantation from November 2004 to July 2007.The symptoms,changes of ankle/brachial index(ABI),limb salvage rate and immediate patent rate were observed. Results PTA/stenting procedure was successful in 51 out of 60 patients(65 limbs)with the technical success rate of 83.3%.Symptoms were improved in 53 cases(88.3%)including complete symptom remission in 40 cases(66.7%),partial remission in 13 patients(21.7%).The procedure failed in 7 cases(11.6%).In successful cases,AB1 increased from preoperative 0.40±0.18 to postoperative 0.91±0.22(P＜0.01).The amputation below knee was performed in two cases and toe apodizers in four cases.The limb salvage rate was 91% during the same hospitalization.Discharged 54 cases were followed up with 14.5±1.2 months,during this period amputation above knee was performed in 2 cases,amputation below knee in 2 cases,and toe apodizers in 2 cases,with a limb salvage rate of 88.9%(48/54).Symptoms were recurrent in five cases,with recurrence rate of 9.2%.Vascular reocclusion or restenosis were found in 10 cases.the patent rate was 81.5% and the cumulative patent rate was 57.3% in one year. Conclusions Endovascular interventional treatment for infrapopliteal arterial ischemic disease is safe and effective.

Objective To evaluate the prevention and management for the complications in interventional therapy for the lower extremity arteriosclerosis obliterans (ASO). Methods In this study 207 ASO cases received interventional therapy. The relationship between complication rates and different TASC type and the influence of diabetes mellitus (DM) with coronary heart disease (CHD) on critical organs complication rates was analyzed. Results The intervention succeeded in 190 cases. There was 17 failures, including 13 procedure failures and 4 deaths due to postoperative critical organ complications. Among the complications there were puncture site hemorrhage in 12, pseudoaneurysm in 4, alimentary tract hemorrhage in 2, arteriorrhexis in 6, cerebral infarction in 8, acute heart failure in 9, respiratory failure in 13, renal failure in 6, thrombosis in 5 and blue-toe syndrome in 1. The intraoperative complication rate of femoral-political type (39.84%, 51/128) was high than that of aorta-iliac type (18.99%, 15/79) (P< 0.05). Those critical organ complication rates of patients with DM and patients with DM&CHD were 27.66% (13/47)and 24.49% (12/49)respectively, they were higher than that of patients without (5%, 2/40) (P<0.05). Conclusions The high complication rates during interventional therapy of ASO were because of misoperation and complicated pathological type and together with DM and CHD. These rates could be decreased when we treated DM and CHD before operation and chosen a reasonable method and shorten the duration of operation.

Objective To evaluate clinical outcome of patients treated with endovascular dilation and stent placement for ilio-femoral venous postthrombotic syndrome (PTS).Methods 42 PTS cases were enrolled in our group from June 2003 to December 2011.Dilation and stent placement in stenosis/ occlusion of ilio-femoral veins were performed in all cases,temporary femoral arteriovenous fistula was established in 24 cases.Results These were not severe perioperative complications.All patients were followed up for 8-75 months.Preoperative limb ulcers in 5 cases were healed.Early thrombosis in stents found in 3 cases was cleared by catheter directed thrombolysis (CDT).Severe in-stent restenosis (＞ 50％)was found in 10 cases,treated with dialation or re-stenting.Primary patency,assisted primary patency and secondary patency rates were 75％,78％,80.1％;64.1％,67.1％,69.6％;63.8％,65.3％,66％;61.7％,65％,65.2％,respectively,at 6 months,12 months,24 months and 36 months.Postoperative Venous Clinical Severrity Score (VCSS) decreased (P =0.000).There was statistical significant improvement in every dimension evaluated by the MOS item short from health survey(SF-36) after operation (P =0.000).Conclusion Ilio-femoral venous stenting is a safe and effective treatment for PTS and with good clinical midterm results.

Objective To study the diagnostic value of thin-slice reconstruction of multi-slice spiral CT(MSCT) in solitary pulmonary nodule(SPN).Methods 55 cases of SPN confirmed by pathology were analysed retrospectively.All cases underwent chest MSCT scan and thin-slice reconstruction images were obtained at 10.0 mm,5.0 mm,2.5 mm and 1.25 mm thickness.The detecting rate of CT findings of SPN on different thickness CT images was evaluated,and the effect of thin-slice images and traditional CT scan in diagnosing benign and malignant pulmonary nodules ?2 test.Results(1)In detecting rate of CT findings of SPN,the thin-slice(1.25 mm) reconstruction was better than traditional CT(P

Objective To explore the impact and value of 11C-MET PET/CT for high-grade glioma in making conformal radiotherapeutic planning compared with MRI.Methods From October 2011 to September 2013,11C-MET PET/CT and MRI were performed in 25 initial glioma cases and 9 recurrent cases.All 34 cases (19 males,15 females) were classified into initial group (n=17),initial surgery group (n=8) and recurrent group (n=9).GTV was outlined using PET/CT images combined with T1 weighted MRI images.Results 11 C-MET PET/CT optimized radiotherapy targets in 22 cases,including 7 initial cases,8 initial surgery cases and 7 recurrent cases.GTV was consistent with the volume of abnormally enhanced region on T1 weighted MRI (VMRI) in 10 initial cases,GTV＞VMRI in 5 cases,and GTV＜VMRI in 2 cases.GTV＜VMR1 in the initial surgery group.GTV was consistent with VMRI in 2 recurrent cases,GTV＞VMRI in 2 cases and GTV＜VMRI in 4 cases; 1 recurrent case's GTV located largely outside the VMRI.Conclusions There are some differences on locating the lesions of high-grade gliomas between 11C-MET PET/CT and MRI.Using 11CMET PET/CT in sketching radiotherapy target could significantly optimize the high grade glioma's radiotherapeutic planning,which may be more valuable in recurrent or operative cases.