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 investigate the effect of intermittent hypoxia on neuronal apoptosis and inducible nitric oxide synthase (iNOS) expression in rat hippocampus,in order to explore the potential mechanism of hippocampal neuronal apoptosis induced by intermittent hypoxia.Methods Twenty-four Wistar male rats (280-350 g) were randomly divided into three groups:the normal control,intermittent normoxia and intermittent hypoxia group (n=8 each).The animal model of intermittent hypoxia was established by an automated nitrogen/oxygen profile system.The three groups were respectively exposed to continuous normoxia (21 ％O2),cyclical normoxia (21 ％O2),and cyclical hypoxia [alternating between normoxia (21 ％ O2) and hypoxia (5 ％ O2),every 120 seconds] throughout the eight hours of light time(8:00-16:00).The rats were dissected and the hippocampus was removed at the end of the designated duration of exposures for six weeks.TdT-mediated dUTP nick end labeling (TUNEL) was used to detect the apoptotic rate of the hippocampus region in each group.Reverse transcription-PCR,immunohistochemistry (IHC) and Western blotting were used to examine mRNA and protein expressions of iNOS in the hippocampus tissue.Results The apoptotic rate of hippocampal neurons was higher in intermittent hypoxia group than in intermittent normoxia group [(28.236±0.081) ％ vs.(9.341±0.026)％,P＜0.05].The mRNA and protein expressions of iNOS were higher in intermittent hypoxia group than in intermittent normoxia group (3.394± 1.344 vs.0.125±0.040,7.793±0.052 vs.1.356±0.039,both P＜0.05].Conclusions Intermittent hypoxia can induce hippocampal neuronal apoptosis in rats,accompanied by the upregulation of iNOS gene transcription and protein expression,which indicates that iNOS may involve in the process of hippocampal neuronal apoptosis induced by intermittent hypoxia.

Objective To study the incidence frequency of prothrombin G20210A (FⅡ G20210A)mutation in the patients with pulmonary thromboembolism(PTE)in northeast China,and to clarify the predictive value of FⅡG20210A mutation detection in PTE of the population in northeast China.Methods 60 PTE patients(PTE group) and 80 sex-matched healthy controls(control group)from the same geographic area were selected.All the patients were diagnosed by lung ventilation/perfusion scan and/or multi-slice CT pulmonary angiography(CTPA)as well as medical history.The genome DNA was extracted from the whole blood using alcohol.Polymerase chain reaction (PCR),restriction fragment length polymorphisms(RFLP)analysis with HindⅢ restriction enzyme and sepharose gel electrophoresis were used to identify the F Ⅱ G20210A mutation in PTE group and control group. Results After digested by HindⅢ restriction enzyme,only the fragments of 407 and 99 bp were found in PTE group.The frequency of FⅡ G20210A mutation was 0%,there was no statistical difference compared with contol group(P>0.05).There were no heterozygote and homozygote mutation of FⅡ G202210A gene in PTE group and control group.Conclusion The incidence of FⅡ G20210A mutation in the PTE patients in northeast China is very low,and the detection of FⅡ G20210A mutation may have no predictive value in PTE of the population in northeast China.

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 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

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.