BACKGROUND: Studies show that riluzole can greatly inhibit protein kinase C-?Ⅱ in mediating vascular endothelial cell growth factor effects on endothelial cells, so we presume it can be used in inhibiting retinal neovascularization. OBJECTIVE: To explore the inhibitory effects of riluzole on the proliferation of human umbilical vein endothelial cells and retinal neovascularization, and investigate potential mechanism of its effects. DESIGN, TIME AND SETTING: Single-sample observation and randomized controlled animal trial were performed at the Animal Laboratory and Basic Study Laboratory of Chongqing University of Medical Sciences from August to December 2007. MATERIALS: Thirty 3-day-old healthy C57BL/6 mice (60 eyes), irrespective of gender, were selected to model proliferative retinopathy induced by hyperoxia. Raw material of riluzole was product of Venturepharm Laboratories Limited, Beijing (No. 060630, content ≥98.5%). METHODS: Human umbilical vein endothelial cells were cultured, and randomly divided into 3 groups. The mice in hyperoxia model group and riluzole group were modeled, and riluzole group was additionally intraperitoneally injected 10 mg/kg per day riluzole; control group was intraperitoneally injected normal saline. MAIN OUTCOME MEASURES: Effect of riluzole on proliferation of endothelial cell proliferation induced by vascular endothelial growth factor was observed using MTT assay; The inhibitory effects of riluzole on retinal neovascularization were evaluated by counting the endotheliocyte nuclei of new vessels beyond the inner limiting membrane in sections with HE staining. The expression of retinal protein kinase C-?Ⅱand vascular endothelial growth factor were detected with SABC immunohistochemistry. RESULTS: The riluzole of 0.1-10 ?mol/L inhibited proliferation of human umbilical vein endothelial cells in a dose-effect dependent manner. The number of the endotheliocyte nuclei of new vessels beyond the inner limiting membrane was obviously less in eyes of normal control and riluzole groups than in the hyperoxia group (P

TCM medical service trade is an important component of TCM service trade. Developing TCM medical service trade has wide market and strategic importance. Development of TCM medical service trade in China is still in the initial exploring stage with small scale and dominated by the government. Recently, foreign patients prefer TCM medical serve. TCM medical service trade in abroad shows the partial and small scaled characteristics. The number of joint venture TCM treatment centers is small, and movement of personnel is rare. In order to promote the development of TCM service trade, this article proposed the following four suggestions: conducting TCM international tele-medicine service trade, developing TCM medical travel, investing more in the development of foreign TCM medical centers, expanding the scale of exporting TCM medical talents.

Objective To investigate the effect of imaging parameters and postprocessing methods on the quality of MR imaging of short T2 components with 3D ultrashort TE (UTE) double echo pulse sequence. Methods 3D UTE double echo pulse sequence was performed on dry human femoral specimen and the tibial diaphyses, knee joints, and tendons of ankles of a group of healthy volunteers. To investigate the effect of different trajectory delays of the imaging system(-6, -3, -2, - 1,0, 1,2, 3 s), different flip angles(4°, 8°, 12°, 16°, 20°, 24°), different TEs (0. 08, 0. 16, 0. 24, 0. 35 ms)and different postprocessing methods(difference imaging of subtracted volume and non-volume UTE)on the 3D UTE MR imaging quality, the SNR and CNR were calculated and compared, and the artifacts of the images were analysed. Results The cortical bone, periosteum, tendon and meniscus showed high signal intensity on the images of UTE pulse sequence. The best SNR was acquired with 2 s trajectory delay. The best flip angle was 8° to 12° for the human UTE imaging in vivo. The highest CNR was obtained from the TE of 0. 08 ms. The longer the TE was, the more artifacts appeared. The SNR of difference imagewas improved when image subtraction was performed afer multiplanar reconstruction (MPR) of the primary double echo images.Conclusions The short T2 components show high signal intensity on the MRI of 3D UTE double echo pulse sequence. The imaging quality can be improved by shortening TE, using appropriate flip angle and performing subtraction for difference image after MPR of the primary double echo images.

An evaluation system including experiment preparation,experiment process,com-prehensive design of experiment,experiment skills and written exam was established in order to adapt to the experimental teaching reform in our school. Experimental preparation assessment was to evaluate‘preview and self-evaluation report’prepared and submitted by students. Experimental process assess-ment was to evaluate students' classroom performance and experiment reports. Assessment of compre-hensive design experiments was to evaluate the overall participation of students. Skill assessment was consisted of oral test and experiment operation test. Final written examination,mainly consisting of subjective questions,emphasized on student's flexible use of knowledge and ability to solve practical problems. The evaluation system of promoting student's learning and teacher's teaching through the examination not only fully arouse student's attention on experiment,but also make teachers more ob-jectively and really understand students' learning situation and the teaching effectiveness.

ObjectiveTo observe the effects of human IL-10 gene transfection on the mRNA and protein expressions of IL-1β and TNF-α in the penumbra area following focal cerebral ischemia-reperfusion injury in rats and to investigate its neuroprotective mechanism. MethodsRats were divided into four groups: normal control group, ischemic control group, empty plasmid group and human IL-10 gene transfected group. The mRNA and protein expressions of IL-1β and TNF-α in the penumbra area were detected by fluorescence real-time quantitative PCR and ELISA respectively. ResultsIn normal control group, ischemic control group, empty plasmid group and human IL-10 gene transfected group, the levels of protein expression of TNF-α in penumbra area were(0.66±0. 04) ,(1.16±0.26),(1. 155±0. 26)ng/g and(0. 84±0. 05)ng/g, and the levels of protein expression of IL-1βin penumbra area were(0.37±0.05), (1.25±0.39), (1.21±0.57) ng/g and(0.62+0.05)ng/g, respectively. Compared with normal control group, the levels of protein expression of TNF-α and 1L-1β were significantly higher in other three groups(all P＜0. 01), and lower in human IL-10 gene transfected group than in ischemic control group and empty plasmid group(all P＜0. 01). In normal control group, ischemic control group, empty plasmid group and human IL-10 gene transfectedgroup, the levels of mRNA expression of TNF-α in penumbra area were 1.00 ±0.53,9.42±1.83,9.69±1.96 and 3.53±1.09, and the levels of mRNA expression of IL-1β in penumbra area were 1.00 ±0.51,27. 81±4.84,23.96 ± 4.90 and 13.55± 4.45, respectively. Compared with normal control group, the levels of mRNA expression of TNF-α and IL-1β were significantly higher in other three groups(all P＜0. 01), and lower in human IL-10 gene transfected group than in ischemic control group and empty plasmid group(all P＜0. 01). ConclusionsThe human IL-10 gene transfection may play an protective effect on cerebral ischemia through inhibiting mRNA and protein expression of IL-1β and TNF-α in the penumbra area following focal cerebral ischemia-reperfusion in rats.

BACKGROUND:The effect of advanced glycation end products (AGEs) on bone resorption is controversial. Our previous study has shown that bone resorption is significantly inhibited when AGEs present with pre-osteoclast cells RAW 264.7, while the effect of AGEs on osteoclastic acidification remains unknown. OBJECTIVE:To investigate the effect of AGEs on osteoclastic acidification and the underlying mechanism. METHODS:RAW 264.7 cells were induced by RANKL (15μg/L;normal group) to generate osteoclasts, and AGEs (50-400 mg/L;experimental group) or bovine serum albumin (100 mg/L;control group) were added at the beginning of the induction. The effect of AGEs on bone resorption was evaIuated by anaIyzing the area of bone resorption on the Osteo Assay Surface plates, and the effect of AGEs on osteoclastic acidification was evaluated by acridine orange staining. Furthermore, the expression levels of V-ATPase a3 and CIC-7 were detected to investigate the underlying mechanism. RESULTS AND CONCLUSION:The bone resorption area in the AGEs group was significantly decreased compared with the normal group (P<0.05). Acridine orange staining reveaIed that the red fluorescence (620 nm) intensity in the AGEs group was significantly decreased compared with the normal group (P<0.05), and this inhibitory effect became obvious with the increase of AGEs concentration. Immunocytochemistry, western blot assay and PCR findings showed that the expression levels of V-ATPase a3 and CIC-7 in the AGEs group were decreased significantly compared with the normal group (P<0.05). To conclude, AGEs exert inhibitory effect on osteoclastic acidification, probably by inhibiting the expression of V-ATPase a3 and CIC-7.

Objective To evaluate the diagnostic value of the cell block (CB) with immunostaining method by endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) biopsy for pancreatic lesions.Methods A total of 72 patients with pancreatic lesions underwent EUS-FNA at the First Affiliated Hospital of Guangxi Medical University from March 2012 to June 2013.The EUS-FNA samples of all patients were processed by conventional smear cytology,liquid-based cytology (LBC) and the cell block with immunostaining.Results There were 61 pancreatic patients who were finally diagnosed as having pancreatic tumors,including 55 cases of pancreatic cancer,2 pancreatic solid pseudopapillary tumor,4 pancreatic endocrine tumors (PETs),and 11 benign lesions:4 chronic pancreatitis,2 pancreatic tuberculosis,4 pancreatitis and 1 pancreatic mucinous cystadenoma.The diagnostic sensitivity of conventional smear cytology,liquid-based cytology and cell block with immuno-staining method were 68.9％ (42/61),75.4％ (46/61) and 90.2％ (55/61),respectively.The diagnostic specificity of three methods were all 100.0％.The diagnostic accuracy rates were 73.6％ (53/72),79.2％ (57/72) and 91.7％ (66/72),respectively.The diagnostic accuracy rate of the cell block with immunostaining was higher than those of conventional smear cytology (P ＜ 0.05) and the liquid-based cytology (P ＜ 0.05).Conclusion Endoscopic ultrasound-guided fine-needle aspiration is a safe and effective method with high sensitivity and specificity in the diagnosis of pancreatic lesions.Cell block method combining immunohistochemistry helps to increase the diagnosis and histological diagnosis of pancreatic lesions.The cell block has a greater clinical value in the diagnosis of pancreatic lesions.

Objective:To explore the effects and mechanism of anti IL-9 antibody on malignant ascites ( MA) of hepatic carci-noma in mice.Methods:A mouse model of MA was established by mouse H 22 cell line.45 mice were divided randomly into experi-mental group,negative control group and blank control group at 24 hours after modeling,with 15 mice in each group.The experimental group was injected intraperitoneally with anti IL-9 antibody;the negative control group was injected with isotype IgG antibody;the blank control group was injected with normal saline .The weight and behavior of the mice were measured before each injection .Five mice of each group was sacrificed at 24 hours after the last injection to measure the volume of MA .The level of VEGF,MMP-2,IL-9 and IFN-γin MA were determined with ELISA assay;the survival time of rest mice were recorded and compared .Results:The mean volume of MA of experimental group,negative control group and blank control group were (6.70±1.52)ml,(10.28±1.75)ml,(10.36±2.30) ml,respectively,the MA volume of experimental group were lower as compared to negative control group and blank control group ( P<0.05).The mean survival time of experimental group was (17.2±2.1)d,which was significantly prolonged compared with the negative control group (14.5±1.2)d and the blank control group (14.8±1.4)d (P<0.05).The levels of VEGF of MA in experimental group was significantly lower compared to the negative control group and blank control group (P<0.05).The levels of IL-9 of MA in experi-mental group was significantly lower compared to the negative control group (P<0.05).The levels of MMP-2 and IFN-γin experimental group had no significant difference compared with the negative control group and blank control group ( P>0.05 ) .Conclusion:Intraper-itoneal injection anti IL-9 antibody on H22 ascites-bearing mice can effectively inhibit the generation of the MA .The mechanism may be related to the inhibition of the expression of the VEGF and IL-9.

Objective:To investigate the effects and mechanisms of interleukin-22(IL-22) on inhibiting liver fibrosis induced by HSC,and explore the role of Wnt/β-catenin pathway in the activation of hepatic stellate cells(HSC).Methods:Rat HSC was activated by TGF-β1,and the mRNA and protein levels of β-catenin and α-SMA were detected by q-PCR and Western blot,respectively.HSC was treated with different hours and concentration of recombinant rat protein IL-22.The cell proliferation rates were detected by CCK8,cell apoptosis rates were tested by flow cytometry.HSC were treated with optimal concentration of IL-22 after activated by TGF-β1,the cell proliferation rates,mRNA and protein levels of β-catenin and α-SMA were compared of before and after intervention.Results:The mRNA and the protein levels of β-catenin and α-SMA were significantly increased after activated by TGF-β1(P<0.05).IL-22 inhibiting the proliferation of HSC in a dose-and time-dependent manner (P<0.05) and decreased the mRNA and the protein expression level of β-catenin and α-SMA(P<0.05),but had no significant effect on apoptosis rates(P>0.05).IL-22 significantly inhibited the activation of HSC induced by TGF-β1 and remarkably decreased the mRNA and the protein expression level of β-catenin and α-SMA(P<0.05).Conclusion:The Wnt/β-catenin pathway may participates in the process of HSC activation and α-SMA secretion,and IL-22 inhibits biological function of HSC in a dose-and time-dependent manner.This effect probably via inhibited the Wnt/β-catenin signal pathway.

Objective To study the immune tolerance effect after renal transplantation,an animal model of tolerant dentritic cell adaptive transfer renal transplantation was constructed.Methods Tolerant Dentritic cells from SD rats were separated and purification in vitro,after a series of procedures,the Dentritic cells were administrated in the portal vein of Wistar rats,then conduct the operation of renal transplantation for Wistar rats.Renal tissue pathology and splenic lymphocyte reaction were test in 3、5、7、9、11、和13d after renal transplantation.Results After the renal transplantation,time of acute rejection happened in control group and experimental group is 4.69?1.26d and 10.69?1.63d,respectively.The mean survive time between two group has significant difference.Conclusion Renal transplantation immune tolerance could be induced by the tolerant dentritic cell of donor,by the mean of administrating the cell into host portal vein.The immune tolerance has specificity.