Objective To study the influence of microenvironment created by dissoluble cytokines derived from hepatocellular carcinoma cells (HCCs) on the functional status of mitochondria in dendritic cells (DCs) in order to investigate the mechanisms of tumor immune escape.Methods CD14+ monocytes,which were isolated with immune magnetic beads from fresh peripheral blood of healthy human,were then cultured in RPMI1640/10% FBS supplemented with 100 ng/ml rhGM-CSF and 100 ng/ml rhIL-4 for 7 d to develop into immature DCs (imDCs).Maturation was induced by addition of 20 ng/ml rhTNF-? to imDCs for another 3 d of culture.Acquired imDCs and mDCs were respectively co-cultured with human HCCs for 48 h in Transwell chamber.These treated cells were investigated for mitochondrial membrane potential,enzyme activity and intracellular Ca2+ concentration ([Ca2+]in) in the cytoplasma by immune fluorescence and MTT assay.Those untreated cells served as control.Results After co-cultured with HCCs,flow cytometry showed that the mitochondrial membrane potentials of imDCs and mDCs were respectively decreased from (659.991?17.052)and(473.741?11.676)to(482.681?7.935)and(407.189?5.051)(P

Objective To study the influence of microenvironment created by dissoluble cytokines derived from hepatocellular carcinoma cells (HCCs) on the expression of IL-12 of dendritic cells (DCs) at different differentiating stage. Methods The CD14+ monocytes,isolated with immune magnetic beads from fresh peripheral blood of healthy human,were cultured in RPMI 1640/10% FBS supplemented with 100 ng/ml rhGM-CSF and 100 ng/ml rhIL-4 for 7 d to develop into immature DCs (imDCs). Maturation was induced by addition of 20 ng/ml rhTNF-? to imDCs for another 3 days' culture. Acquired imDCs and mDCs were respectively co-cultured with human HCC Bel7402 cells for 48 h in Transwell chamber. The concentrations of IL-10,IL-12,TGF-?1 and VEGF in culture supernatant were investigated by ELISA,imDCs and mDCs cultured alone served as control. Results After co-cultured with Bel7402 cells,IL-12 were decreased from (115.076?8.129) pg/ml to (17.599?1.757) pg/ml in imDCs,and from (258.346?3.609) pg/ml to (6.787?1.123) pg/ml in mDCs (P

Objective To evaluate the immune reconstruction of patients with advanced gastric cancer treated by autologous peripheral blood stem cells following chemotherapy. Methods 54 patients with advanced stage of gastric cancer were accepted arterial chemotherapy under the guidance of DSA with the regimen of high dosage of EAP (VP16 100 mg/m 2, topmycine60 mg/m 2 and carplatine200 mg/m 2). Among them 20 patients underwent autologous peripheral blood stem cell transplantation after 48 hours of the chemothrapy. Results Effective rate of EAP arterial chemotherapy combined with autologous peripheral blood stem cell transplantation was up to 85.0% (17/20) in advanced gastric cancer, 29.3% (10/34) in control group ( P

Objective To evaluate the role of mierosurgery in surgical treatment to urethrocutaneous fistula after urethroplasty in hypospadias and improve surgical results.Methods From 1999 to 2006,44 urethrocutaneous fistulae (more than 3mm in diameter) after urethroplasty for hypospadias in 33 patients were repaired with different skin flaps.For example,Thiersch technique,urthroplasty,etc.Microsurgical tech- nique was employed in every case.Results The success rates of different procedure were 84.8% (28/33) for Thiersch technique,100% (11/11) for urethroplasty respectively.The total success rate was 88.6%(39/ 44).Conclusion It's just application of skin flap for repairing of big or complex urinary fistula after hypos- padias surgery.The application of microsurgical technique can increase success rate.It is necessary to excise scar and partial urethra for hypospadias fistula combined with urethral structures,cicatricial eontracture and in- curvation of penis.Rich blood-supply,low tension and atraumatic technique are all very important to improve surgery success rates of urinary,fistula after hypospadias repair.

Objective To evaluate the therapeutic effect of pedical conjunctival flap transposition on refractory corneal ulcer. Methods The data of 56 eyes of 56 patients with refractory corneal ulcer who underwent pedical conjuctival transposition in a tertiary eye unit in China from June 2005 to June 2009 were retrospectively analyzed. There were 36 infective corneal ulcers, 9 keratoplasty graft ulcers, 4 traumatic ulcers, and 7 chemical burn ulcers. All patients were treated with focal debridement and pedical conjuctival flap transposition. Results Forty-seven out of 56 (83.9%) were cured after one surgery, and 5 (8.9%) had conjunctival flap retraction, among which 3 underwent the second surgery and 2 had the third surgery to completely heal. Four patients (7.2%) turned to lamellar keratoplasty because of fungal corneal ulcer relapse. Ulcer in 52 patients healed and the globe preserved. They had different levels of vision improvement. Conclusion Pedical conjunctival flap transposition is effective for refractory corneal ulcer.

Objective To investigate the effect of miR-210 on angiogenesis of human renal clear cell carcinoma . Methods Detect the expression of miR-210 in 40 cases of renal clear cell carcinoma tissues , and analyze the rela-tionship between the expression of miR-210 and microvessel density ( MVD) .miR-210-ASO was lipotransfected in-to renal clear cell carcinoma cell line 786-O.RT-qPCR was used to verify the transfection effect .The effect of the supernatant of control group , negative control group and miR-210-ASO group tumor cells on the lumen formation of human umbilical vein endothelial cells ( HUVEC) was observed in a 3-D culturesystems .The transplantation tumor model of nude mice was established , and the effect of miR-210 on the formation of the transplanted tumor micro vessel was observed by endomucin and VEGF immunofluorescence staining under laser scanning confocal micro -scope.Results The expression of miR-210 was positively correlated with microvessel density in renal clear cell carcinoma ( P<0.05 ) .The expression of miR-210 was significantly decreased in 786-O cells transfected with miR-210-ASO( P<0.05 ) .The lumen formation of HUVEC cells co cultured with miR-210-ASO group cell supernatant was significantly less than that of control group and negative control group ( P<0.05 ) .The tumor volume of miR-210-ASO group was less than that of the control group , and the number of the micro vessel and the VEGF expres-sion were significantly less than that of the control group ( P<0.05 ) .Conclusions Inhibition of miR-210 can suppress blood vessel formation in renal clear cell carcinoma .

BACKGROUND: The problem to be solved firstly in adipocyte implantatior after its in vitro adherent ulture f preadipocyte with scaffold using tissue-engineered technique is the biocompatibility of scaffold and dipocyte.OBJECTIVE: To investigate the feasibility of poly-β-hydroxybutyric acid (PHB) used for readipocyte implantation.DESIGN: Randomized controlled trial.SETTING: Medical College of Wuhan University.MATERIALS: This trial was carried out in Medical College, Wuhan University during October 2003 o June 2005. Twelve male SD rats(Experimental Animal Center, Medical College, Wuhan University), weighing rom 200 to 250 g, were involved.PHB scaffold was provided by Institute of Polymer Materials, Department f hemical Engineering, Tsinghua University.METHODS: The preadipocytes of rats were isolated, purified and ultured in vitro for use. PHB scaffold was made into 0.75 cm×0.75 cm ×0.2 cm lamellar cell biological caffold, then which were soaked by DMEM/F12 culture medium containing 150 g/L fetal bovine serum and laced in culture plate, one piece each well, 12 wells totally. 1.0 mL cultured cell suspension was added n each well to prepare cell scaffold complex. Eight rats were selected. The prepared complexes were implanted into right-side back tissue of a rat and fixed with 5-0 silk, serving as adipocyte scaffold roup; the biological scaffolds without adipocyte adherence were implanted into left-side back tissue of he same rat with the same method, serving as blank scaffold group. One lamellar biological scaffold was mplanted into each side. Eight weeks later, the appearance and structural change of grafts were observed, nd the grafts were weighted and their volumes were measured. Four lamellar biological scaffolds were mplanted into the other 4 rats separately and taken out after 3 weeks, then they were fixed by 100 g/L eutral formalin, sliced, and stained by haematoxylin and eosin (HE). Their histological changes were observed.MAIN OUTCOME MEASURES: ① Gross and histological observation of grafts in adipocyte scaffold roup and blank scaffold group. ② Comparison of volume and mass of grafts in two groups.RESULTS: Twelve ets were involved in the result analysis ,without deletion. ①Gross observation: In the adipocyte scaffold group, grafts presented flesh-color appearance and peplos in the peripheral region, and newly ormed minute blood vessels ingrowing into the scaffold. In the blank scaffold group, grafts presented rey ppearance, which was coated by peplos. ② HE staining: Three weeks later, a few scattered adipocytes were ound in the peripheral region of the grafts,there were very few vessels. Eight weeks later, lamellar ibrous peplos was found around the complex. Adipocytes ingrew from the peripheral region of scaffold into caffold, partial scaffold was nearly full of adipocytes and many vessels formed;In the blank scaffold roup, only peplos and vessels were found, scaffold was full of fibrous connective tissue and adipocytes ere not found. ③ Comparison of volume and mass: adipocyte scaffold group was superior to blank group in scaffold volume and mass [(257.5±70.2)vs.(144.6±62.6)mm3,(245.6±58.2) vs.(148.7±60.3)mg, both P＜0.01].CONCLUSION: Preadipocyte can adhere to PHB scaffold, proliferate and differentiate into mature dipocytes. It is feasible for PHB material to serve as a carrier for preadipocyte.

Objective To discuss the surgical outcomes of atrial septal defects(ASD) by closing the ostium secundum via small chest incision.MethodsTotally 41 cases with ASD were treated by occlusion via a small incision(3-4 cm) at the right anterior chest.After cutting the pericardium,the right atrial was sutured with double ring and a special occluding device was inserted to close the ostium.Afterwards,under the guidance of transthoracical or transesophageal echocardiography,the satisfiable position for the occluder was confirmed by repeated testing with the protect string.After that,the right atrial and the occluder were sutured by a needle thread for fixation.ResultsOf the patients,the procedure was successfully completed in 39 cases.The mean operation time was 45 to 95 minutes(mean,60 minutes).The patients were discharged from our hospital in 3 to 6 days.No dislocation of the device or atrial shut was found in 2 to 24 months after the operation.Two patients were converted to open surgery because of failure in the occlusion.ConclusionOcclusion via small chest incision is safe,minimally invasive,and convenient procedure for ASD.

Objective To study the effect of continuous positive airway pressure(CPAP) therapy for 56 clinical cases neonatal a-cute respiration prostration. Methods Based on the traditional treatment,CPAP assistant therapy were taken on 56 cases. The pressure of CAPA were adjusted according to the variations of clinical condition and blood gas value. Results After 6 - hour assistance of CPAP performance, the symptom of all cases had been abated, pa (O2) was increased, the normal CPAP applaying time was 28 hoours. The remarkable difference had been found compared to those in no CPAP practice(P