The potamon that were sold by Mengla market were examined positive for two species of paragonimus metacercariae andexcyted metac ercariae, this is the first report in Yunnan Province． The potamon that were sold by cities market from other suburbs may cause epdemic or outbreak of paragonimiasis, there is of momentous significance in epidemiology．

Objective To explore the long-term efficacy of botulinum toxin type A injections on facial beauty and its long-term safety.Methods A total of 33 beauty-seekers with botulinum toxin type A treatment for more than five years were reviewed as observation group,using digital muscle palpation meter (Myoton PRO) for the determination of the orbicularis oculi muscle and masseter muscle tension (F),the muscle characteristic parameters,such as muscle hardness (S) using a homemade facial questionnaire test for satisfactory rate of beauty from both beauty-seekers and physicians.At sametime,33 normal adults that never accepted botulinum toxin injection with matched age and gender were collected as control group.The same-sex indicators were dtermined and compared with using statistic analysis t test.Results The pairwised parameters of the same sex and site were comparied between the two groups;average F and S values in the observation group were lower than those of the control group,but no statistically significant difference were observed between the two groups (P＞0.05);in the observation group,average appearance age was 7.3 years younger than the control group,and the facial shape improved significantly.Conclusions Long-term and repeated application of botulinum toxin A is able to remove crow's feet and decrease the masseter and so the injection is safe with high satisfaction to beauty-seekers.

ObjectiveTo study the effect of SSY-B2 on regeneration of central nervous system of rats with bilateral fornix/fimbria transaction.MethodsMale adult SD rats were divided randomly into 6 groups as sham group,model group, positive control agent piracetam group, SSY-B2 low dosage(1.5g crude drug/kg) group, medium dosage(3g crude drug/kg) group and high dosage(6g crude drug/kg) group.The bilateral fornix/fimbria transection in the rats were carried out. After operation, drugs were fed introgastrically to each group respectively for 6 weeks. The immunoreactive products of growth-associated protein-43(GAP-43 )and chondroitin sulfate proteoglycans(CSPG) in defined areas were measured using immunohistochemical methods. ResultsThere was no difference in number of cells expressing GAP-43 between the model group and sham group (P>0.05),but that in low dosage group increased compared with the model group (P<0.001). The CSPG in parietal lobes after lesion expressed,which was in sham group and model group(sham group and model group respectively, 56.43±59.6,116.36±10.561), and SSY-B2 in low and medium dosage inhibited its expression compared with the model group (P<0.05,P<0.01). Conclusions SSY-B2 can enhance the expression of GAP-43 and inhibit the expression or deposition of CSPG, promote axonal regeneration in the CNS, and thus structural repair and functional restoration in certain degree.

ObjectiveTo evaluate the possible effect of SSY-B2 on reducing the loss of neurons and enhancing the expression of nerve growth factor(NGF).MethodsMale adult SD rats were divided randomly into 6 groups as sham group,model group,positive control agent piracetam group, SSY-B2 low(1.5g crude drug/kg), medium(3g crude drug/kg)and high dosage (6g crude drug/kg)group.Bilateral fornix/fimbria transection was carried out in the rats' septohippocampal pathway and 6 weeks' drug treatment was administered with different doses of SSY-B2 and positive control agent piracetam. After behavioral tests, the numbers of neurons in medial septum and immunoreactive products of NGF in different areas were measured, using Nissle staining and immunohistochemical methods.ResultsThere was neural loss in medial septum after fornix /fimbria transection, but SSY-B2 at each dosage markedly reduced the loss(59.13±22.02,50.60±23.18,63.93±18.35,the number of neurons for three SSY-B2 dosage groups,P<0.005 for all compared with the model group 20.33±14.01).The number of NGF positive cells decreased in model group, but did not show significant statistic difference compared with the sham group (P>0.05) in the medial septum, polymorph layer of dentate gyrus and entorhinal cortex/Subiculum area. In the medial septum, all three dosage enhanced the expression of NGF positive cells(145.1±57.7,161.3±08.2,200.6±58.2,the number of neurons for three SSY-B2 dosage group,P<0.005 for all compared with the model group 50.2±48.6). SSY-B2 at low and medium dosage group also increased the number in both entorhinal cortex/Subiculum and polymorph layer of dentate gyrus.Conclusions SSY-B2 can reduce the loss of neurons in the medial septum, which may be involve in increasing expression of NGF;NGF expression in dentate gyrus, subiculum of hippocampal formation and entorhinal cortex increased by SSY-B2 may play a role in the compensation of these area for learning/memory.

ObjectiveTo observe the effect of SSY-B2 on microglial cells in rats after bilateral fornix/fimbria transection.MethodsMale adult SD rats were divided randomly into 6 groups as sham group,model group,positive control agent piracetam group, SSY-B2 low(1.5g crude drug/kg), medium(3g crude drug/kg)and high dosage (6g crude drug/kg)groups. Half to 1 hour before operation, water or drugs were fed introgastrically to each group respectively and continued for 6 weeks.The tissues of brain was gained and the immunoreactive products of BS-I B4 (Isolectin B4 from Bandeiraea Simplifolia, a marker for microglia) in the perilesion area was measured using immunohistochemical methods.ResultsThe number of microglia of the sham and model groups was (30.3±21.8) and (114.5±102.3) respectively, P<0.05. That of three different dosage of SSY-B2 groups was(249.7±149.4), (252.0±191.7)and (244.2±154.9), P<0.05 for each group compared with the model group.ConclusionMicroglia number in the perilesion area can be increased by SSY-B2, which may contribute to the nerve repair and functional improvement after injury.

BACKGROUND:Clinical trials have shown that oral administration of valsartan can decrease in-stent restenosis after stent implantation.But whether valsartan used locally also has the sanle effect and the possible mechanism should be validated.OBJECTIVE:To observe the effect of valsartan-eluting stents on collagen deposition in neointima and AT2 receptor expression after implanting valsartan-eluting stents into rabbit abdominal orta.DESIGN:Randomized and controlled animal experiment.SETTING:Beijing Friendship Hospital.MATERIALS:The experiment was performed at the Laboratory of Beijing Friendship Hospital between October 2004 and March 2006.Fifteen New Zealand white rabbits,irrespective of gender,weighing 2.75-3.25 kg were selected(Animal Laboratory of Beijing Friendship Hospital).The rabbits were adaptively fed for one week.All the operations of rabbits during the experiment were accorded with animal ethical standards.Valsartan powder was presented as a gift by Novartis.China;Reagent of MASSON was provided by Department of Pathology of Beijing Friendship Hospital;1%picrosirius solution was provided by the Department of Pathology of China-Japan Friendship Hospital:Mice-anti-rabbit monoclonal AT2 antibody was product of Santa Cruz Biotechnology (USA);Envision reagent was purphased from Dako;primers were synthesized by SBS Genetech(SBS).METHODS:①The animals were randomized into bare-metal stent group,carrier-eluting stent group and valsartan-eluting stent group with 5 animals in each group.All rabbits were implanted with corresponding types of above-mentioned stents into abdominal aortas down below renal artery.②Quantitative angiography before,immediately after and 3 months after stent implantation were performed to compare vascular diameters of the aortas.③Three months Iater,the rabbits were executed after anaesthesia.The vessels with stents were processed with HE staining.Indices of the vascular neointimal formation,I.e. iBrier and external elastic membrane luminal area,the maximal intimal thickness,neointimal area and stenosis area percent were measured.④The collagen deposition in neointima was observed through MASSON staining,and the type of collagen was identified through picrosirius stain.⑤The expressions of AT2R mRNA and proteins were also compared by RT-PCR and immunohistochemistry among three groups.MAIN OUTCOME MEASURES:①The diameters of aorta with stent at different time;②Inner and extemal elastic membrane luminal area,the maximal intimal thickness,neointimal area and stenosis area percent;③Collagen deposition and type of collagen of the aorta with stent;④AT2R mRNA and protein expressions.RESULTS:Of 15 rabbits selected in the experiment,1 rabbit of the bare-metal stent group died during stent implanting,and 1 of the carrier-eluting stent group died during breeding after stenting.Finally,13 rabbits were included in final analysis.①There were no significant differences in the mean aortic diameters between any two of the three groups before,immediately after and 3 months after stent implantation(P＞0.05).②A larger 1uminal area and a less neointimal hyperplasia in valsartan eluting-stents group were found compared with the other two groups(P＜0.01).③MASSON staining showed that collagen deposition was rich in neointima of bare-metal stent group and carrier-eluting stent group while rare in neointima of valsartan eluting stent group.Pierosirius staining suggested that the deposited collagen was type Ⅲ collagen predominantly accompanied by type Ⅰ collagen around stents struts;the type Ⅲcollagen deposition was obviously decreased in valsartan eluting stent group.④AT2R protein only expressed in adventitia of bare-metal stet group and arrier-eluting stent group while expressed in all layers of valsartan eluting-stents group.The AT2R mRNA/a-Actin mRNA of valsartan eluting stent group was significantly higher than that in the other two groups(P＜0.01).CONCLUSION:Valsartan eluting-stents inhibits neointimal hyperplasia after stenting by decreasing collagen deposition.especially collagen Ⅲ.The mechanism may be related with the upregulation of AT2R mRNA and protein expressions by valsartan-eluting stent.

AIM: In order to study the relationship between mitochondrial deficiency and Alzheimer's disease(AD), we used sodium azide, a specific inhibitor of cytochrome C oxidase(COX), to develop a cell model of mitochondrial complex IV deficiency and investigated the impairment of microtubules and microtubule-associated proteins. METHODS: Primary cultured hippocampal neurons of hewborn rats were exposed to sodium azide ,then cell viability was measured by MTT method; cell morphology, immunofluorecence-stained cellular microtubules and microtubule-associated proteins were observed by confocal microscopy. RESULTS: Primary cultured hippocampal neurons were exposed to 8-128 mmol/L sodium azide for 3-24 h, MTT absorbance decreased dose-and time-dependently. Exposed to 64 mmol/L sodium azide for 6 h, the processes of cells retracted, synapses disappeared, axons were shortened under contrast microscope. Meanwhile, microtubles were disassembled and became disorderly, the expression of microtubule-associated proteins were also reduced especially in the processes observed by confocal microscopy. CONCLUSIONS: Sodium azide inhibits the assembly and polymerization of tubulin in microtubules which may be reduced by low expression of microtubule-associated proteins in nerve cells. The damage of axons induced by microtubule collapse further blocks the intercellular signal transduction and intracellular material transportation which are important causes in cell death.

Objective To measure the expressions of programmed death?1(PD?1)and programmed death ligand?1(PD?L1)on peripheral blood T lymphocytes of patients with condyloma acuminatum(CA), and to investigate their role in cellular immunity in these patients. Methods Peripheral blood samples were obtained from 30 patients with CA(CA group)and 20 healthy human controls (control group). Flow cytometry was conducted to detect the expressions of PD?1 and PD?L1 on the surfaces of peripheral blood CD4+and CD8+T lymphocytes, and to determine the counts of CD4+and CD8+T lymphocytes. Enzyme?linked immunosorbent assay(ELISA)was performed to measure the levels of serum interleukin?2(IL?2)and interferon?γ(IFN?γ). Statistical analyses were carried out to compare the above parameters between the two groups, and to assess the relationship of PD?1 and PD?L1 expressions with the counts of CD4+and CD8+T lymphocytes as well as with the serum levels of IL?2 and IFN?γ. Results There was a significant increase in the expression rates of PD?1 and PD?L1 on CD4+T lymphocytes(PD?1:9.48%± 3.31%vs. 7.12%± 2.16%, t=2.81, P<0.01;PD?L1:4.40%± 1.46%vs. 3.26%± 1.13%, t=3.16, P<0.01)and CD8+T lymphocytes(PD?1:12.52%± 3.17%vs. 9.95%± 2.17%, t=3.16, P<0.01;PD?L1:7.07%± 2.23%vs. 5.39%± 1.69%, t=2.88, P<0.01)in the CA group compared with the control group. Moreover, the CA group showed significantly lower counts of CD4+T lymphocytes(727.43 ± 138.59/μl vs. 804.25 ± 92.83/μl, t=2.17, P<0.05)and CD4/CD8 ratio(1.23±0.35 vs. 1.46 ± 0.34, t = 2.24, P < 0.05) than the control group, while no significant difference was observed in CD8 + T lymphocyte counts between the CA group and control group(613.60 ± 121.60/μl vs. 572.45 ± 103.08/μl, t=1.24, P>0.05). The levels of serum IL?2 and IFN?γwere both lower in the CA group than in the control group(t=2.12, 2.16, respectively, both P < 0.05). In the CA group, PD?1 and PD?L1 expression levels on peripheral blood CD4 + T lymphocytes were both negatively correlated with CD4+T lymphocyte counts, the CD4/CD8 ratio, as well as IL?2 and IFN?γserum levels(all P<0.05), and those on peripheral blood CD8+T lymphocytes were also negatively correlated with the CD4/CD8 ratio(all P<0.05), but uncorrelated with CD8+T lymphocyte counts(both P>0.05). Conclusion PD?1 was highly expressed on peripheral blood T lymphocytes from patients with CA, which may inhibit T lymphocyte?mediated immune response, decrease CD4+T lymphocyte counts, the CD4/CD8 ratio as well as IL?2 and IFN?γserum levels by interacting with its ligand PD?L1 and forming the PD?1/PD?L1 signaling pathway.

AIM: Gene transduction with a recombined murine stem cell retroviral vector has been investigated to find an effective way of gene transduction and to offer theory and experimental basis for the recombined murine stem cell retroviral vector used for gene transduction. METHODS: 1. Construction of retrovirus vector: EC1-4 gene (repeats 1-4 of cadherin-5 extracellular domain) and mutant (Ser 222A) MEK1 gene were cloned into retrovirus vector pMSCV after cut by Bgl Ⅱ and EcoR 1 restriction endonuclease. 2. Obtaining CD41 + cells and cell culture: Cells expressing CD34 + from cord blood were isolated. The inducement of cells expressing CD41 from CD34 + cells was performed by using TPO and cells CD41 + were selected by FACS. NIH 3T3 cells were cultured in high sugar DMEM medium and U937 in RPMI 1640 medium. UT7 cells which is cytokine-dependent cell line were grown in Iscove's modified Dulbeco's medium supplemented by GM-CSF. 3. Determination of viral titers: Retroviral vectors were transferred to packaging cell line 293. Retroviral containing supenatant was collected after transfection. The viral titers was tested on infection of NIH 3T3 cells by FACS analysis. 4. Western blot: Transduced CD41 +,UT7,U937 and MDA-MB-435 cells were analyzed by western blot to examine expression of transduced genes. RESULTS: A packaging cell line 293 produces high-titer MEK1 pMSCV retroviruses (3.1?10 7) and EC1-4 pMSCV retroviruses (1.0?10 8). With 8-folds dilution retroviruses,60.73% GFP positive cells have been obtained in MEK1 pMSCV transduced UT7 cells,72.56% in U937 cells and 30.57% in CD41 + cells,respectively. GFP positive cells have reached up 97.54 % in EC1-4 pMSCV transducted MDA-MB-435 cells. Phosphorylated MEK1 has been decreased in experiment group when TPO has stimulated CD41 + and UT7 cells or serum has stimulated U973 cells. This indicates that is a dominant negative effect of mutation MEK gene. EC1-4 gene transduced MDA-MB-435 cells have expressed EC1-4. CONCLUSION: The recombined murine stem cell retrovirus can effectively mediate gene transduction of CD41 +,UT7,U937 and MDA-MB-435 cells,and transduced genes can be stably expressed.

Aim To investigate the effects of tyrosine kinase inhibitor A77-1726 on IL-13 induced STAT6 phosphorylation and c-fos expression in Dami cell and to provide novel experimental basis to the clinical application of A77-1726 and the study of IL-13 pathway.Methods Total RNA was extracted from Dami cells incubated with or without IL-13 and A77-1726 respectively for various time points.RT-PCR and agar gel electrophoresis were used to examine the expression of c-fos mRNA.The expression of STAT6 and c-fos proteins was detected by Western blot.A densitometric rel-ative quantitation of PCR and Western blot products was quantitated using Image Quant software.Results STAT6 was phosphorylated by treatment of 100 ?g?L-1 IL-13 in Dami cells.Phosphorylation of STAT6 induced by IL-13 was inhibited by treatment of 50?mol?L-1 A77-1726.The expression of c-fos mRNA was significantly induced by IL-13 treatment in Dami cells(P