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.

[ABSTRACT]OBJECTIVE To obser ve t he pathological changes of nasal mucosa in rats after instillation of 37℃ normal saline. METHODS Forty-eight qualified rats were given instillation of 37℃ nomal saline 30 times, once per minute. Another 2 qualified rats served as control. After instillation, in the 15, 30 minutes and 1st, 3rd, 7th, 14th, 42th, 28th day respectively, the nasal septum mucosa of 6 rats was observed by light and electron microscopy. The corresponding area of the mucosa of control group was also observed by light and electron microscope. RESULTS Under light microscope, the arrangement of cell was disordered in the 1st, 3rd, 7th days. But there is no damage of vascular gland structure. After 14 days, the disorder recovered. With electron microscope, edema of cells, expansion of perinuclear gap, disorder of cilia and microwilli was found in the 30 minutes, 1st, 3rd day. All these began recover in the 7th days, and completely restored in the 28th day. CONCLUSION 37℃ nomal saline drip can damage nasal mucosa, but the damage is light, which is characterized by disorder and shortage of cilia and microvilli, edema of the epithelial cells. And the damage can recover quickly. It began in the 30 minutes after instillation, reached its peak in the 3rd day. And the recovery began in the 7th day, and completed in the 28 day.

Objective To investigate the upper airway caliber and the corresponding pharyngeal wall in obstructive sleep apnea syndrome(OSAS) by spiral CT.Methods Seven patients with OSAS confirmed by polysomnography(PSG) underwent spiral CT scan.The thickness of posterior pharyngeal wall of retropalatal region(RP),retroglossal region(RG) and epiglottal region(EPG) were measured.Results ①Among 7 patients,5 had only RP obstruction,and 2 had RP and RG regions obstruction;②The length and width of soft palate were 44.28 mm?1.16 mm(normally 41.90 mm?1.95 mm) and 9.42 mm?0.46 mm(normally 5.84 mm?0.24 mm) respectively;③In OSAS patients,the thickness of posterior pharyngeal wall of RP,RG and EPG region were statistically larger than that of the normal group.Conclusion Spiral CT is of practical value in evaluating the pharyngeal airway obstruction of OSAS.

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 explore the value of maternal serum triple markers screening,consisted of AFP (α-fetal protein),β-human chorionic gonadotropin (β-hCG) and free estriol (uE3),for Down's syndrome,in second trimester.Methods We searched published literatures from PubMed,MEDLINE,China National Knowledge Internet (CNKI) and Wanfang Database from January 1990 to August 2014 and articles met the following criteria were collected:(1) The report was of a screening test;(2) The research purpose was to study the efficiency ofAFP,hCG and uE3 (triple markers) screening for Down's syndrome;(3) The research subjects were pregnant women in second trimester;(4) All of the studied cases were confirmed by amniocentesis and chromosome karyotyping;(5) The pregnant outcomes must be available;(6) Each report should have at least one case of Down's syndrome identified;(7) The literature could be retrieved by Science Citation Index or Chinese core periodicals;(8) When assessed by QUADAS (quality assessment of studies of diagnostic accuracy included in systematic reviews) Quality Assessment Scale,the score should be ≥ 8.Information were extracted,including name of the first author,publication time,sample size,sensitivity,specificity,maternal age,gestational age,cutoff value,β-hCG type and others.MetaDiSc 1.4 software was applied for meta-analysis.I2 was used for heterogeneity,and the fixed or random effects model for calculation of the combined sensitivity,specificity,diagnostic odds ratio (DOR) and the 95％CI.The summary receiver operating characteristic (SROC) curve was drawn.Deek's test in Stata Software was used to validate publication bias.Results A total of 49 literatures were recruited in this study with a total sample size of 960 245.Deck's funnel plot analysis showed that the correlation coefficient and the standard error of bias were-1.067 and 3.64 (t=-0.290,P=0.771).The correlation coefficient and the standard error of the slope were 3.578 and 0.26 (t=13.740,P=0.000).The random effects model showed the pooled sensitivity of the 49 literatures was 0.72 (95％CI:0.70 0.74),the pooled specificity was 0.92 (95％CI:0.92-0.93),and DOR was 33.80 (95％CI:25.03-45.65).The area under the SROC was 0.900 7.DOR of younger age group (including three literatures) was 14.38 (95％CI:3.67-56.42) and 26.64 (95％CI:19.49-36.41) for the older age group (two literatures).For two literatures determining the gestational age based on ultrasonography,the DOR was 50.22 (95％CI:26.91-93.71),and DOR was 33.09 (95％CI:17.33-63.19) for those based on last menstrual period in these two literatures.For eight literatures applied the cutoff value of 1:270,12 applied 1:250 and four applied mixed cutoff value,the DOR was 10.94 (95％CI:3.04-39.38),54.34 (95％CI:42.19-70.01) and 36.37 (95％CI:31.19-42.40),respectively.DOR of total β-hCG group (12 literatures) was 22.06 (95％CI:16.46-29.58) and that of free β-hCG group (ten literatures) was 37.15 (95％CI:30.00-46.02).Conclusions Triple regimen of second trimester maternal serum screening for Down's syndrome is much more efficient.The detection rate could be further improved by determination of the gestational age with ultrasound,and application of 1:250 as the risk cutoff value and free β hCG as a screening marker.