A new teaching reform project in experiment of Human Parasitology is performed by improving experiment content and adopting a new teaching method.The project elucidates that teaching reform in experiment can enhance students'abilities in the experiment,self-teaching and problem-solving and problem-analyzing,and creative ability.

Objective To investigate the effect of microbubbles mediated ultrasound insonation on proliferation and apoptosis of vascular smooth muscle cells (VSMCs) in different phase of cell cycle. Methods Rat thoracic aortic VSMCs were cultured in vitro by the method of tissue adherence. The cells were synchronized by the methods of serum starvation and double thymidine block. The synchronization results were detected by flow eytometer. VSMCs in different phases of cell cycle were exposed to 1 MHz continuous waves ultrasound for 120 s at intensity 0.3 W/cm2 in the presence of lipid-coated microbubbles (1 ml/L). Apoptosis of VSMCs was analyzed by AnnexinV/PI staining using flow eytometry. The proliferation and the proliferating cell nuclear antigen(PCNA) protein expression of VSMCs were detected by MTT assay and immunoeytochemistry, respectively. Results The synchronized G0/G1 and S phase VSMCs were achieved, with synchronized rates to 89.53 % and 66.87 %, respectively. Ultrasound sonication for 120 s with microbubbles could significantly inhibit the proliferation and downregulate the PCNA expression of S phase VSMCs,but the proliferation and PCNA expression of G0/G1 phase VSMCs were not affected. After treatment of ultrasound with microbubbles, the apoptotic ratio were found to reach (7.05 ± 2.04)% in G0/G1 phase VSMCs and (27.01 ±3.87)% in S phase VSMCs. Conclusions Microbubbles mediated ultrasound insonation can significantly inhabit the proliferation and induce apoptosis in VSMCs at proliferation stage.

Objective To investigate the effect of ultrasound irradiation combined with liposome membrane microbubbles on the reorgnization of cytoskeleton in vascular smooth muscle cells (VSMCs). Methods Rat thoracic aortic VSMCs were cultured in vitro. VSMCs were exposed to 1 MHz continuous waves ultrasound radiation for 120 s at intensity 0.3 W/cm2in the presence of liposome membrane microbubbles (1 μl/ml) after treated with platelet derived growth factor-BB (PDGF-BB). The reorganizations of microfilaments, microtubules and intermediate filaments were examined by using immunofluorescence and fluorocytochemistry techniques. Results There was a substantial increase in the expression of F-actin and assembly of long bundles of stress fibers in the transversed cell body when treated with PDGF-BB. Neither alterations of β-tubulin nor of vimentin cytoskeletal protein organization were observed in PDGF-BB treated cells as compared to those of the contol group. After ultrasound irradiation combined with liposome membrane microbubbles, the expression of F-actin, β-tubulin and vimentin were reduced along with the simultaneous changes in microfilaments, microtubles and intermediate filaments array. Conclusions Ultrasound irradiation combined with liposome membrane microbubbles can induce significant changes in cytoskeleton structure of VSMCs cultured in vitro.

0.05) was found between the dechlorinated water group and the extract of A.philoxeroides group in the activities of LDH and SDH after 12 h or 20 h of exposure.Conclusion The extract of A.philoxeroides rapidly inhibits ChE,and then the activity of Mg2+-ATPase,which may suppress the release and utilization of ATP in the Oncomelania snails and finally causes death of snails.

This study is aimed to establish a high-performance liquid chromatography (HPLC) method for simultaneous determination of skimmin, scopolin and umbelliferone in Saussurea hieracioides. Samples were analyzed on a Wondasil C18-WR column (4.6 mm x 250 mm, 5 microm) with methanol (A) and water containing 0.1% phosphate (B) as mobile phases for gradient elution at a flow rate of 1.0 mL x min(-1). The detection wavelength and column temperature were set at 325 nm and 35 degrees C, respectively, and the sample size was 10 microL. The results showed that skimmin, scopolin and umbelliferone were simultaneously achieved within 40 min under the above conditions. A good linearity was observed in the range of 0.18-5.6 microg (r = 1.000 0), 0.060-1.8 microg (r = 0.999 9), 0.032-0.97 microg (r = 0.999 8) for skimmin, scopolin and umbelliferone, respectively, with the average recoveries of 99.16% (RSD = 0.41%), 100.3% (RSD = 0.79%), 102.2% (RSD = 0.87%). The method is simple, accurate and reproducible and can be used for the quality control of S. hieracioides.
Chromatography, High Pressure Liquid ; Coumarins ; analysis ; Glucosides ; analysis ; Medicine, Tibetan Traditional ; Reproducibility of Results ; Saussurea ; chemistry ; Umbelliferones ; analysis

OBJECTIVE:To observe clinical efficacy and safety of tanshinone combined with limbal stem cells transplantation in the treatment of pterygium. METHODS:Totally 97 cases (118 eyes) of primary pterygium admitted into our hospital during Feb. 2010-Sept. 2014 were analyzed retrospectively,and divided into observation group (48 cases,57 eyes) and control group (49 cases,61 eyes). Both groups received autologous limbal stem cell transplantation. The control began to give Tobramycin and dexamethasone eye drops 1-2 drop 1 week before surgery,every 4-6 h one times. Observation group was given Tanshinone cap-sules 0.5 g,po,tid,one week before surgery,for 3 months. Repair time of corneal epithelium and local symptom regression time were compared between 2 groups. Corneal astigmatism and corrected visual acuity were observed in 2 groups before and 1,3 months after surgery. The occurrence of recurrence and ADR was analyzed statistically in 2 groups. RESULTS:The repairing time of corneal pithelial and local symptom regression time in observation group were significantly shorter than control group,with statis-tical significance (P<0.05). There was no statistical significance in corneal astigmatism and corrected visual acuity between 2 groups before and one month after surgery (P>0.05). 1,3 months after surgery,corneal astigmatism of 2 groups was decreased significantly and corrected visual acuity was increased significantly than before surgery,and 3 months after surgery the observation group was significantly better than the control group,with statistical significance (P<0.05). The recurrence rate of observation group was 3.51%,which was significantly lower than 14.75% of control group,with statistical significance(P<0.05). There was no statistical significance in the incidence of ADR between 2 groups(P>0.05). CONCLUSIONS:Tanshinone combined with autol-ogous limbal stem cell transplantation in the treatment of pterygium can shorten the time of corneal epithelial repair and local symp-toms,restore the visual function of patients and reduce recurrence rate with good safety.

Objective To assess the clinical diagnosis value and treatment effect of anti-Müllerian hormone(AMH)and inhibin B(INHB)in polycystic ovary syndrome(PCOS)patients.Methods Total of 300 cases of PCOS patients were enrolled in this study from January 2014 to January 2016 in the First Affiliated Hospital,Hunan University of Chinese Medicine,and those patients were randomly divided into group A,group B and group C.There were 100 patients in every group.The patients in group A were interfered by traditional Chinese medicine.The patients in group B were treated with Western medicine and those in group C were treated with traditional Chinese medicine combined with western medicine.Total of 264 cases health volunteers were enrolled as the control group.The effect was evaluated.The level of AMH and INHB in serum of PCOS patients were detected by chemiluminescent assay before treatment and three months after treatment.Results The cutoffs of AMH and INHB were 6.98 ng/ml and 150 pg/ml,respectively.The AUC of AMH combined with INHB was significantly larger than that of AMH or INHB(0.945 vs.0.859,0.945 vs.0.784).In the PCOS group,the positive PCOS rate of AMH combined with INHB was significantly larger than that of AMH or INHB[87.00％(261/300)vs.83.33％(250/300)vs.93.67％(281/300),x2=15.593,P=0.000].The sensitivity[93.67％(281/300)],specificity[92.42％(244/264)],positive predictive value[93.36％(281/288)],negative predictive value[92.78％(244/264)]and Jordanian index(0.659)of AMH combined with INHB was significantly larger than that of AMH[87.00％(261/300),87.88％(232/264),89.08％(261/293),85.61％(232/271)and 0.612]or INHB[83.33％(250/300),90.15％(238/264),90.58％(250/276),82.64％(238/301)and 0.571].After treatment,AMH[(9.06±2.13)ng/ml vs.(6.34±1.12)ng/ml,t=10.595,P=0.000;(9.08±2.08)ng/ml vs.(6.02±1.02)ng/ml,t=13.209,P=0.000;(9.13±2.31)ng/ml vs.(3.53±0.83)ng/ml,t=22.814,P=0.000]and INHB[(173.13±14.22)pg/ml vs.(145.26±13.05)pg/ml,t=14.440,P=0.000;(174.28±13.82)pg/ml vs.(145.39±12.98)pg/ml,t=15.238,P=0.000;(174.98±13.77)pg/ml vs.(133.15±12.04)pg/ml,t=22.869,P=0.000]in 3 groups had decreased.After treatment,the AMH of group C [(3.53±0.83)ng/ml] was significantly lower than that of group A and B[(6.34±1.12)ng/ml and(6.02±1.02)ng/ml,F=237.936,P=0.000],and the level of AMH in group C [(133.15±12.04)pg/ml] was significantly lower than that in both group A and group B[(145.26±13.05)pg/ml and(145.39±12.98)pg/ml,F=30.645,P=0.000].Conclusions AMH combined with INHB can be used to diagnose PCOS.AMH and INHB can be used to evaluate PCOS efficacy.

Objective:To analyze the correlation between the glycosylated hemoglobin and immunologic function in patients with type 2 diabetes.Methods:Clinical data of patients with type 2 diabetes received treatment at our hospital from 2011 to 2014 was analyzed (Group A) and also included the healthy control as Group B.Results:A total of 75 patients were analyzed,Group A 45 cases and Group B 30 cases.Group A had a higher level of glycosylated hemoglobin than that of Group B , the difference was statistically significant(8.12±3.45 vs 4.87±1.65;t=4.472,P<0.001).Group A patients had lower levels of IgA,IgM and CD4/CD8 when compared with Group B ,the difference was statistically significant ( P<0.05 ).Correlation analysis shown the glycosylated hemoglobin had significantly negative correlation with IgA ,IgM and CD4/CD8 ( P<0.05 ).Conclusion: Patients with type 2 diabetes have a low immune function which are related to high level of Glycosylated hemoglobin.

Objective To investigate the effect of insulin-like growth factor-1(IGF-1) and basic fibroblast growth factor (bFGF) on the differentiation of human dental pulp stem cells in vitro. Methods Mono-cell suspension was separated from the human adult dental pulp with collagenase Ⅰ and dispase digestion. Colony-forming efficiency of cells was calculated. Cells were observed under an invert microscope. Dentin sialoprotein (DSP), dentin matrix protein 1 (DMP-1) and dentin sialophosphoprotein (DSPP) mRNA of the 3rd passage cells induced with IGF-1 and bFGF in vitro were detected by immunohistochemistry and RT-PCR. Results Clonogenic cells were obtained from dental pulp and colony-forming efficiency of cells was 2.6-3.5 colonies/10 4 cells. Human specific DSP, DMP-1 protein and DSPP mRNA expressed in colony-forming cells after 7 d of induction with IGF-1 and bFGF in vitro. Conclusion Adult stem cells might exist in human dental pulp. Dental pulp stem cells could differentiate into odontoblast under the induction of IGF-1 and bFGF.

Objective To investigate the effects of tazarotene on the proliferation of cultured human keratinocytes and IFN-?-induced expression of HLA-DR in those cells.Methods Keratinocytes were cul-tured from normal human skin in vitro,and were treated with various concentrations of tazarotene(10 -5 ,10 -6 ,10 -7 mol/L).At24h,48h after treatment,the effects on cell proliferation were assessed by MTT method.The expression of HLA-DR was determined using immunocytochemistry techniques in cultured human ker-atinocytes incubated with tazarotene,IFN-?or both for24h.Results①The proliferation of keratinocytes was decreased when exposed to10 -7 -10 -5 of tazarotene as compared to non-exposed keratinocytes after24h and48h.Moreover,the effects on cell proliferation by tazarotene were dose-dependent;②There was rare expression of HLA-DR in normal human keratinocytes.③HLA-DR expression was inducible significantly with500u/mL of IFN-?,but failed to be induced with10 -6 mol/L of tazarotene,in keratinocytes at24h af-ter treatment.④After24h combined treatment of10 -7 -10 -5 mol/L of tazarotene and IFN-?,the induction of HLA-DR expression was significantly stronger,in a dose-dependent manner,than IFN-?alone(P