The effect of trichosanthin on mast cell degranulation and histamine release were observed with both Alcian blue staining method and fluorescent histamine determination method. It was showed that, ① in vitro trichosanthin showed no direct effect on the mast cell degranulation and histamine release even in the presence of fresh serum; ② in vivo administration on of trichosanthin in mice could cause significant mast cell degranulation and histamine release in inflammatory situ, as shown by the decreased number of Alcian blue stained nondegranulated mast cell in the peritoneal cavities; ③ intra peritoneal injection of trichosanthin in Wistar rats could significantly increase histamine level in the whole blood, while inducing prominent exudation of protein into the peritoneal cavities; ④ the supernatant of peritoneal exudate lavaged 12 hours after trichosanthin injection could cause mast cell degranulation. These results implicate that, although it showed no direct effect on mast cells, trichosanthin could cause mast cell to degranulate and release histamine via in vivo activation of some soluble mast cell degranulating factors. It was discovered in further study that the depletion of complement by cobra venom factor injection revealed no effect on the production of mast cell degranulation factor, suggesting that, in normal animals, the trichosanthininduced complement activation had no significant influence on the mast cell degranulation and histamine release.

BACKGROUND:Human-mammal chimeric liver chimera has been a vital significance for the proliferation and differentiation of bone marrow mesenchymal stem cells.
OBJECTIVE:To establish an animal model of human-rhesus chimeric liver using adult bone marrow mesenchymal stem cells.
METHODS:Adult bone marrow mesenchymal stem cells were isolated, purified and cultured for the sixth generation. The number of bone marrow mesenchymal stem cells was no less than 5×108. Bone marrow mesenchymal stem cells labeled with green fluorescent protein were transplanted into the liver of the embryo rhesus with pregnancy of 10 weeks under guided by type-B ultrasound. At the 1st and 3rd months of birth, the liver tissue of the infant rhesus was taken for biopsy. After routine pathological section, histological specimens were observed under fluorescence microscope to confirm if there were adult bone marrow mesenchymal stem cells positive for green fluorescent protein and their distribution, and detected by immunohistochemical staining to identify if human albumin expressed in the liver of infant rhesus.
RESULTS AND CONCLUSION:Fluorescence microscope observation indicated that at the 1st and 3rd months after birth, there were surviving bone marrow mesenchymal stem cells derived from human with green fluorescence in the liver of infant rhesus, and these cells migrated to form more concentrated distribution. The immunohistochemical results demonstrated that functional liver cells expressing human albumin were observed in the liver of infant rhesus at the 1st and 3rd months after birth, and their distribution was in accordance with bone marrow mesenchymal stem cells with green fluorescence. Human-rhesus chimeric liver can be established using adult bone marrow mesenchymal stem cells, which can generate functional liver cells in the liver of infant rhesus.

Objective To explore the clinical utility of fluorescence in situ hybridization (FISH) urine test as a non-invasive method for diagnosing the upper urinary tract urothelial carcinoma (UUTUC).Methods Urine specimens from 30 consecutive patients with UUTUC were analyzed by means of FISH and cytology.Ultrasonic and CT were also used to analyze urine specimens from the study group.Urine specimens from 30 patients with other diseases were also analyzed by means of FISH and cytology in order to compare the specificity.Results The sensitivities of FISH,cytology,CT and ultrasonic were 87％,37％,90％ and 43％.The specificity of FISH and cytology were 97％ and 93％.The sensitivity between FISH and MMCT were compared and they were not statistically significantly superior to ultrasonic and cytology.FISH and cytology were compared and no specific statistical significance was found.The positive and negative predictive value for FISH was 96％ and 88％.For cytology it was 85％ and 60％.Conclusions FISH analysis is a useful ancillary test in the detection of UT-TCC with excellent sensitivity and specificity.It provides a more reliable and less invasive approach to diagnosis and postoperative follow-up for UT-TCC.

Objective To investigate the efficacy of retroperitoneal laparoscopic heminephroureterectomy for duplex kidney anomalies.Methods Retroperitoneoscopic heminephroureterectomy was performed on nine patients, six males and three females.The average age of the study group was 37 years ( range 13 to 58).Seven cases had anomalies on the upper kidney pole, two cases had anomalies on the lower kidney pole.Five anomalies were on the left side, two were on the right side and two were in bilateral sides (one special case had three ureters on the left side and two ureters on the right side ).Three cases complained of flank pain; two cases were found hydronephrosis by physical routine examination;Three cases complained of flank pain and fever; one cases complained of hematuria and kidney atones.All the cases were preoperatively diagnosed by color doppler ultrasound, MRU, IVP or CTU.Retroperitoneal laparoscopic heminephroureterectomy was performed on all patients.The operation time, blood loss, hospital stay, intraoperative and postoperative complications and efficacy were observed.Results All the retroperitoneal laparoscopic procedures were successfully completed.No intraoperative complications were found.The average operation time was 87 min (range, 65 to 125).The average blood loss was 112 ml (range, 30 to 600).The recovery times of intestinal function was 1.6 days ( range, 1 to 3 ).The average postoperative hospital stay was 7 days (range, 5 to 12).The syndrome disappeared and kidney function was normal at a mean followup of 18 monthes.Conclusions Retroperitoneal laparoscopic surgeries for duplex kidney has the benefits of being minimally invasive, fewer complications, quick recovery and certainty of efficacy.Retroperitoneal laparoscopic surgeries can be considered as a first operation method to treat duplex kidney anomalies.

ObjectiveTo investigate the efficacy of transurethral resection and ball pouch dilatation for treatment of ureterostenosis.MethodsThe clinical data of 49 cases of ureteral stricture were analyzed retrospective analysis from June 2008 to June 2011 with 20 cases of male patients and 29 cases of female patients.The age was 15 to 56 years,with a mean age of 40 years.Ipsilateral renal function were mild impairment in 4 cases,moderate impairment in 35 cases,and severe damage in 10 cases.There were ureteropelvic junction etenosis in 11 cases,upper ureteral stricture in 13 cases,and lower segment stenosis in 25 cases.The ureteral stricture length was 0.3 to 2.0 cm,with a mean of 1.2 cm.Seventeen patients were treated with transurethral resection and ball pouch dilatation by minimally invasive percutaneous nephrostomy,and 31 cases were completed by ureteroscopy.The ureteral stents were removed by ureteroscope after 3 - 6 months.45 cases were followed up for 12 -43 months,with a mean of 24 months. ResultsForty-eight cases were completed smoothly with 1 case converted to open surgery.The surgical time was 25 to 95 min with a mean of 42 min.The postoperative hospital stay was 2 to 6 d with a mean of 4 d.In the follow-up of 45 cases,B ultrasound and CT scan showed hydronephrosis reduced significantly in 39 patients,IVU showed unobstructed ureter without significant stenosis.And 6 cases showed no significant changes in hydronephrosis. Conc(t)usionThe method of transurethral resection and ball pouch dilatation has good clinical effect,less pain and shorter hospital stays,which provides a new and effective treatment for patients with ureteral stricture.

Objective:To analyse the change of morphology and internal air flow in upper airway by the use of oral appliance(OA)in patients with obstructive sleep apnea hypopnea syndrome (OSAHS).Methods:A 46-year-old male patient with OSAHS accepted CT scan before and three months after use of OA.Computational fluid dynamics(CFD)model was built on the base of CT scans by Mimics 10.01 and ANSYS ICEMCFD14.0.The internal flow of upper respiratory tract was simulated by ANSYS-FLUENT 14.0 and the re-sults was analyzed by ANSYS-CFD-Post14.0.Results:The most narrow area of upper airway was located in the lower bound of pha-ryngopalatiae,and it augmented from 0.119 4 cm2 to 0.409 9 cm2 after wearing OA;the maximum air velocity was decreased from 11 . 087 m/s to 8.204 m/s,the minimum negative pressure was decreased from -83 Pa to -59 Pa,the resistance of cavum pharyngis de-creased from 250 Pas/L to 145 Pas/L.Conclusion:Application of OA may expanse the upper respiratory tract,decrease the negative pressure and resistance of the upper respiratory tract in narrow area,reduce the collapsibility of the upper airway and maintain the pa-tency of the airflow.

Objective To investigate the effect of transforming growth factorβreceptor typeⅠ(TGFBRⅠ) on adipocyte differentiation by using a small interference RNA (siRNA). Methods The siRNA targeting TGFBRⅠwas synthesized as experimental group, and negative control siRNA was used as control group. The efficiency of TGFBRⅠdepletion and the expression levels of adipocyte-specific transcription factors CCAAT enhancer binding protein α (C/EBPα),peroxisome proliferator-activated receptor gamma (PPARγ) and adipocyte marker gene fatty acid binding protein 4 (FABP4) were detected by quantitative real-time PCR. After treating with adipocyte differentiation agents for 5 days, the cells were stained with oil red O, and the staining of adipocyte was observed and photographed by laser confocal microscope. In addition, with isopropanol extracted oil red O, optical density values of oil red O were measured at a wavelength of 520, and which were compared between groups. Results After transfection of TGFBRⅠ siRNA, gene expression levels of TGFBRⅠwere significantly reduced in ST2 cells, the number of differentiated adipocytes was significantly increased, and the mRNA levels of adipocyte specific transcription factor C/EBPαand PPARγand adipocyte marker gene FABP4 were enhanced compared with those of control group. After treating with adipocyte differentiation agents for 5 days,the number of lipid droplets of cells with transfection of TGFBRⅠsiRNA was increased than that of cells with transfection of control siRNA. The value of optical density was higher in cells with transfection of TGFBRⅠsiRNA than that of control siRNA group. Conclusion TGFBRⅠsiRNA can effectively facilitate adipocyte formation, which suggests that TGFBRⅠis an important regulator of adipogenic differentiation from progenitor cells.

Objective To evaluate the changes in the expression of annexin A2 (ANXA2) in lung tissues in rats with hepato-pulmonary syndrome.Methods Healthy 3-4-month-old Sprague-Dawley rats of both sexes,weighing 220-250 g,were randomly divided into 3 groups:control group (group C,n =10),sham operation group (group S,n =10) and hepatopulmonary syndrome group (group HPS,n =20).The rats were anesthetized with intraperitoneal 3％ pentobarbital sodium 0.1 ml/100 g.Hepatopulmonary syndrome was produced by chronic ligation of the common bile duct.After 5 weeks,the rats were sacrificed and lungs were removed for determination of ANXA2 and smooth muscle actin α (SM-α-actin) mRNA and protein expression in lung tissues by RT-PCR and Western blot,respectively.Results There was no significant difference in the expression of ANXA2 and SM-α-actin mRNA and protein between groups C and S (P ＞ 0.05).The expression of ANXA2 and SM-α-actin mRNA and protein was significantly higher in group HPS than in groups C and S (P ＜ 0.05).Conclusion The expression of ANXA2 in lung tissues is up-regulated in rats with hepato-pulmonary syndrome.

Objective To establish a method for culture of rat pulmonary capillary pericytes.Methods Six male Sprague-Dawley rats,aged 6-7 weeks,weighing 200-220 g,were anesthetized and the chest was opened.The pulmonary capillary was isolated by type Ⅰ collagenase digestion and micropore filtration and cultured in highglucose DMEM/F12 1∶1 containing 10％ fetal bovine serum and 0.5％ mixture of penicillin and streptomycin.The morphology and growth of cells were observed with inverted phase contrast microscope.The positive cells of αsmooth muscle actin (α-SMA),desmin,neuron-glial antigen 2 (NG2),cluster of differentiation 31 (CD31) were counted by immunofluorescence.The percentage of positive cells was calculated.Results The microscopic examination showed cells of shuttle shape or star shape,mononuclear cells,binuclear cells occasionally,oval nucleus,rich cell plasma,growth in the shape of vortex or fence,and no contact inhibition.The percentage of positive cells ofα-SMA,desmin,NG2,and CD31 was (99.0± 1.2)％,(96.0±2.1)％,(99.0±0.7)％ and0,respectively.Conclusion The culture method for rat pulmonary capillary pericytes is successfully established.

Objective To investigate the role of serine threonine protein kinase-1 (Akt1) and Smad3 in lung tissues in development of hepatopulmonary syndrome in rats.Methods Seventy-two healthy male SpragueDawley rats,aged 3-4 months,weighing 200-250 g,were randomly divided into 3 groups (n=24 each):control group (C group),sham operation group (S group) and common bile duct ligation (CBDL) group.The rats were anesthetized with 3％ pentobarbital sodium 45 mg/kg.In group CBDL,laparotomy was performed,the common bile duct was ligated and then the abdomen was closed,while the common bile duct was only exposed,but not ligated and then the abdomen was closed in group S.At 1st,3rd and 5th weeks (T1-3),8 rats were chosen randomly in each group and blood samples were obtained from the abdominal aorta for blood gas analysis.The rats were then sacrificed and lungs were isolated to detect the expression of Aktl and Smad3 mRNA and protein in lung tissues (by RT-PCR and Western blot).The lung tissues were sliced and stained with hematoxylin eosin for examination of the pathological changes of pulmonary capillaries.Results Compared with C and S groups,the expression of Akt1 and Smad3 mRNA and protein in lung tissues was significantly up-regulated at T2,3,and alveolar-arterial oxygen tension difference was increased at T3 in CBDL group (P ＜ 0.05).The pulmonary capillary was obviously dilated at T3 in CBDL group.Conclusion The expression of Akt1 and Smad3 in lung tissues is up-regulated,which may be one of the mechanisms of development of hepatopulmonary syndrome in rats.