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.

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 establish a polarographic method for detecting fluoroacetamide.Methods On the condition of strong basic solution with high temperature fluoroacetamide was decomposed to form a decomposed product ALC-La-F - complex.In a pH4.90buffer solution containing KNO 3 and hexamethylenetetramine,the decomposed product was determined with com-plex-adsorptive wave at-0.67V by polarography.Re sults The detection limit and linear range of the method was2.0?10 -8 mol/L and5.0?10 -8 -1.2?10 -6 mol/L respectively.The regressive equation was y=3.49x+0.042with a correlation coefficient r=0.9996.The method had been applied to the determination of fluoroacetamide in wheat flour samples satisfyingly.Conclu-sion This method was simple and suitable for the determination of fluoroacetamide in flour.

Objective:To investigate the effects of different impl an t diameter on stress distribution in implant bone interface. Method: The stress and strain in implant bone interface for implants with di fferent diameter (3.0 mm,3.75 mm and 5.0 mm), and with 1.47 N loading mesiodist ally were analyzed by 3 dimesion finite element analysis method. Re sults:The Von mises stress (MPa)at cervix of the implants with the diameter (mm) of 3.0, 3.75 and 5.0 were 0.807,0.533 and 1.080;the alveolar bone displacement (?m) at the cervix of the implants 0.232,0.163 and,0.111 respe ctively. Conclusion: The implant with 3.75 mm diameter is more suitable for orthodontic anchorage.

Objective To investigate the reconstructive method with better sensory and erectile function for partial penile defect. Methods The procedure was as follows: dissecting the suspensory ligaments and separating part of crus,the local fat flap was transferred to fill the front space of symphysis ossium pubica.Various pedicle skin flaps were designed as covering materials.Penis residual stump was advanced to anterior portion of the newly reconstructed penile body as “glans”.The post-operative follow-up included taking full layer skin of penis from the patients to observe neurofibril regeneration by light microscope, electron microscope and scanning electron microscope;determining sensation recovery by physical examination;and checking erectile function with NEVA system. Results 42 patients underwent the above mentioned procedures.In these cases,the length of the penis varied from 0.5～4.0 cm in flaccid,1.5～5.0 cm in erect state before operation to 5.0～8.0 cm in flaccid,7.0～12.5 cm in erect state after operation.28 patients were followed up for more than 1 year.Of them 23 men got married with acceptable sexual activity;in them 18 had their children.The microscopic examination showed that the tenuous wavy peripheral nerves were well differentiated and no denaturation and necrosis were observed in the dermis,near subcutaneous adipose and around eccrine glands.The positive granules could be seen in the cytoplasm of regenerative neurofibrils by immunohistochemistry technique.The regenerative neurofibrils and related tissues could be seen under scanning and transmission electron microscope.The penis presented normal pain,touch feeling and 2-point identification ability in more than 12 months after surgery.Over 12 months after surgery,the patient had gained 4～9 events during 8～12 hours of sleep, and the durations were 4 to 37 min;the volume changes were from 114% to 505% over baseline. Conclusions With this method,the reconstructed penis tends to have better appearance and functions.For penile partial defect it is superior to the conventional operation.

Objective To evaluate the usefulness of imging techniques in diagnosis of corticosteroid-induced avascular necrosis of the femoral head(ANFH).Methods Twenty seven patients with avascular necrosis of the femoral head due to long-term corticosteriod treatment were analysed retrospectively.There were 16 femal and 11 male,ranged in age from 20 to 46 years(mean age 35.6 years).The course of disease was 1～5 years.The administration of corticoid orally was 60～25 mg/d taken on 6 months to 5 years.ANFH was appeared 3 months to 2 years after the administration of corticoid.The imaging examinations included radiography in 20 cases,MRI in 15 cases,both radiography and MRI in 10 cases and CT in 8 cases.8 cases were confirmed by operation and pathology.Results 7 femoral heads in 6 cases with early-ANFH were diagnosed by X-ray,21 femoral heads in 15 cases with early-ANFH were diagnosed by MRI and 11 femoral heads in 7 cases were by CT,19 femoral heads showed a typical “line sign”on MRI.Conclusion The “line sign” on MRI is regarded as the characteristic finding in early-ANFH.MRI is the most sensitive method in the early-diagnosis of the corticosteroid-induced ANFH and it can provide important evidence for clinical management.

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.