Objective To investigate the effect of simvastatin on the mobilization and migration of endothelial progenitor cells(EPCs).Methods EPCs were harvested from the bone marrows of two rabbits,cultured with M199,and identified by immunohistochemistry.The identified EPCs were then treated with simvastatin with different concentrations(0,0.01,0.1,1.0 ?mol/L),and their migration induced by simvastatin was determined with Transwell chamber assay.Six rabbits models of cranial bone defect were established and divided into control and experiment groups(3 in each).In order to elicit the effects of simvastatin on mobilization of EPCs,simvastatin was embedded in polylactic acid compound,and implanted into the cranial bone defect area in the experiment group.Meanwhile,polylactic acid was implanted in the control animals.After 10 days,the expression rate of CD34+/CD133+ EPCs in the rabbit peripheral blood was counted by flow cytometry to determine the motivating effect of simvastatin.Results In Transwell experiment,16 hours after adding simvastatin(0,0.01,0.1 or 1.0 ?mol/L),the cell migration ability was obviously increased showing a dose-dependent trend(OD value:0.077?0.014 in control group and 0.075?0.013 in 0.01 ?mol/L group vs 0.097?0.011 in 0.1 ?mol/L group and 0.099?0.019 in 1.0 ?mol/L group,P

Objective To investigate the effect of simvastatin-polylactic acid compound on critical calvarial defects in rats.Methods Twenty male SD rats(150 g?10 g),were used to establish critical cranial defect(10 mm in diameter)model.The animals were randomly divided into control and experiment groups(10 in each).In the control group,40 mg of polylactic acid were implanted into the defect area;whereas in the experiment group,simvastatin-polylactic acid compound were used(20 mg simvastatin and 40 mg polylactic acid).Four and eight weeks after the implantation,the defect area of the rats was observed by X-ray and toluidine blue staining.Results Eight weeks after the operation,X-ray examination showed high-density regions in the defect area in the experiment group,while low-density regions in the control group.The radiopacity of cranial defects were 27.33%?2.54% in the control group,and 74.63%?2.42% in the experimental group(n=5,t=-30.148,P=0.000).Toluidine blue staining showed a few new bone tissues at 4 weeks and fully filled bone defect at 8 weeks in the experiment group.Meanwhile,in the control group,only a small quantity of new bone tissue could be seen on the edge of the cranial defects.Conclusion Locally implanted simvastain-polylactic acid compound is a promising method for the treatment of bone defect owing to its high osteogenic ability.

The enrollment scale of clinical medicine postgraduates with professional degree was expanded each year in order to meet the needs of the society.How to train high quality medical talents in line with modem medical education and people's health needs was the problem we were confronted with.The affiliated hospital of Inner Mongolia medical university lunched reform on training mode of clinical medicine postgraduates with professional degree; the concrete measures included increasing postgraduate pre-service training,implementing tutor responsible system,applying PBL teaching and focusing on the overall quality training.Students' clinical competence was improved after the reform.

Objective To report a case of psoriasis vulgaris associated with acute myelogenous leukemia(AML)(type M4EO).Methods Clinical data from the patient were collected.Histopathologic examination,and examination of bone marrow and peripheral blood smear were performed.The immunologic types of bone marrow cells were analyzed with FACS.Chromosome and G-banding analyses were carried out with cultured bone marrow cells.Results A33-year-old woman had a history of chronic plaque psoriasis for20years.Her cousin had the same disease history.The patient was treated with various therapeutic regi-mens,most of which were traditional Chinese medicines.Recently the patient suffered from myalgia and chest bone pain,periodic bleeding on gums,fever and so on.The abnormal infantile monocytes and promye-locytes were found with bone marrow smear,and crassitude basophilic granules were noticed in eosinophils.The diagnosis of acute myelogenous leukemia type M4EO was made.The diagnosis was confirmed with the immunologic analysis of born marrow cells with FACS.Chromosome and G-banding analyses revealed her karyotype of46,XX,inv(16)/47,XX,inv(16),+8(2/22).The plaque lesions of psoriasis were regressed after allogeneic bone marrow transplantation and the symptoms of AML were resolved.Conclusion It is the first case report of psoriasis vulgaris associated with acute myelogenous leukemia M4EO which responded to allogeneic bone marrow transplantation.

Objective: To investigate the prevalence and influencing factors of comorbidity of chronic diseases among hypertensive patients with uncontrolled blood pressure in Huzhou City, so as to provide insights into community hypertension control. Methods: Hypertensive patients with uncontrolled blood pressure at ages of 35 to 74 years were sampled using a cluster random sampling method from 5 districts (counties) of Huzhou City. Participants' demographics, living behaviors, and development of chronic diseases were collected using questionnaires, and the height, body weight, waist circumference and blood pressure were measured. Blood glucose, blood lipid and other biochemical parameters were detected, and the number and combination of comorbidity of chronic diseases were descriptively analyzed. Factors affecting the comorbidity of chronic diseases were identified using a multivariable ordinal logistic regression model. Results: A total of 1 215 respondents were included, with a mean age of (60.83±7.76) years, and including 652 men (53.66%) and 563 women (46.34%). The prevalence of dyslipidemia, diabetes, hyperuricemia and cardiac encephalopathy was 45.10%, 30.95%, 23.05% and 5.10%, respectively. The prevalence of comorbidity of chronic diseases was 69.22% among respondents, and there were 497 respondents with one comorbidity (40.91%), 272 with two comorbidities (22.39%) and 72 with three and more comorbidities (5.93%). Hypertension+dyslipidemia (20.74%), hypertension+diabetes+dyslipidemia (9.96%) and hypertension+diabetes+dyslipidemia+hyperuricemia (4.36%) were predominant comorbid combinations. Multivariable ordinal logistic regression analysis showed that participants with overweight (OR=1.782, 95%CI: 1.390-2.286), obesity (OR=2.411, 95%CI: 1.802-3.222), grade 2 hypertension (OR=1.438, 95%CI: 1.077-1.919) had a higher risk of multiple comorbidities than those with normal body mass index and controlled blood pressure, and women (OR=0.563, 95%CI: 0.456-0.696) had a lower risk of multiple comorbidities than men. Conclusions The prevalence of comorbidity of chronic diseases was 69.22% among community hypertensive patients with uncontrolled blood pressure in Huzhou City, and the comorbidity of chronic diseases mainly included dyslipidemia and diabetes. Men, overweight, obesity and hypertension resulted in a high risk of comorbidity of chronic diseases.

Objective To study the relationship of T, B and NK lymphocytes with the pathogenesis of chronic urticaria. Methods Flow cytometry was applied to assess the proportion of T, B and NK lymphocyte subgroups in the peripheral blood of 51 patients with chronic urticaria and 30 sex and age-matched human controls. The CD4:CD8 ratio was calculated. Moreover, the symptoms, disease course and response to antihistamines of these patients were evaluated by one physician. Results The percentage of CD8+ T and NK cells, CD4:CD8 ratio were (27.20±8.22)%, (21.20±10.84)% and 1.48±0.62, respectively, in these patients,(29.9±3.74)%, (17.5±3.56)%, 1.24±0.27, respectively, in the controls; the differences were significant between the two groups (all P＜0.05). Decreased levels of CD3+ T cells, CD8+ T cells and B cells were noted in patients resistant to antihistamines compared with those responsive to antihistamines[(61.81±11.70)% vs (75.74±2.36)%, (24.00±7.79)% vs (34.22±9.30)%, (10.78±2.07)% vs (15.25±4.10)%, P＜0.05, 0.01, 0.05, respectively)], while the CD4:CD8 ratio and percentage of NK cells were increased in antihistamine-resistant patients compared to those in antihistamine-sensitive patients [1.67±0.76 vs 1.17±0.41, (28.61±12.62)% vs (12.78±6.02)%, both P＜0.01 ]. In these patients with chronic urticaria, the percentages of CD3+ T and CD8+ T cells were negatively correlated with the symptom scores (R = -0.31, -0.28, respectively, both P＜0.05 ), while the percentage of B cells was positively correlated with the symptom scores and disease course (R = 0.53, 0.55, respectively, both P＜0.01 ). Conclusions There is an abnormality in the proportion of T, B and NK lymphocyte subgroups in patients with chronic urticaria,which indicates that humoral immunity may be involved in the pathogenesis of chronic urticaria and the mechanism for responsiveness to antihistamine.

Background and purpose:Hepatocellular carcinoma suppressor gene-1(HCCS1)is a potential hepatocellular carcinoma supressor gene,and it also plays an important role in sorting of some cytoplasmic proteins.Its carcinoma suppressor function may be related to its sorting function.So it is important to identify the minimum region of functional sequence in HCCS1 that related to the transportation.Methods:The expression vectors containing various lengths of HCCS1 gene were constructed and transfected into HeLa cells mediated by liposomes.The localizations of different HCCS1 fragments and the colocalizations of M6PR with various truncated HCCS1 were determined by laser scanning confocal microscopy,respectively.Results:10 different expression vectors containing various lengths of HCCS1 gene were successfully constructed,it had been shown that the polar localization of HCCS1 protein and the co-localizations with M6PR disappeared when HCCS1 gene was truncated to 1 120 bp from the 3'end by laser scanning confocal microscopy.Conclusions:We identified the minimum localization of the HCCS1 functional sequence that related to the transportation.

Background and purpose:Wuxing soup is popular for it's anti-cancer effect in folk medicine and deserved to be further studied by modern scientific methods.This research aimed to explore its anti-cancer effect and to study the influence on the immunity of melanoma in mice. Methods :Inhibition of different ingredients and concentrations of Wuxing soup on the growth of the mouse melanoma B16 cells was detected by MTT in vitro.Animal experiment was performed to determine its anti-cancer effect in vivo.C57/BL6 mice were randomly divided into three groups after vaccination of B16 cell,and then given intragastrically with different soups for 25 days.All mice were killed on day 26 after inoculation.The weight of tumors were recorded.The anti-tumor immune function was measured by T lymphocyte transforming assay and NK killing assay.The different effect of different ingredients was also observed. Results :Our result showed that different ingredients soup exerted different inhibition on B16 cell growth and Wuxing soup was the strongest one of all and in dose-dependent manner in vitro.The animal experiment indicated that different ingredients soup has different inhibition on melanoma,the soup-treated mouse display improved T lymphocyte transforming assay and NK killing ability.Among the different soups,Wuxing soup showed the strongest anti-cancer effect and immune enhancement. Conclusions :Wuxing soup is an effective anti-cancer agent in melanoma mice and can enhance the immunity of the mice with melanoma.

Objective To investigate the protective effect of brain-derived neurotrophic factor (BDNF) on neuronal injury induced by ropivacaine (Rop) and its mechanism.Methods (1) Experiment one:0,1,2,3,4 and 5 mmol/L Rop was used to stimulate SH-SY5Y cells for 48 h to induce neuronal injury;the morphological changes of the cells were observed under microscope;MTT assay was used to detect the cell activity;flow cytometry was used to detect the cell apoptosis,and immunohistochemistry was used to detect the expressions of protein kinase B (Akt) and proliferating cell nuclear antigen (PCNA).(2) Experiment two:SH-SY5Y cells were treated with 0,1,3,5,7 mmol/L Rop,respectively;the cell activity was measured 48 h after Rop treatment;the semi inhibitory concentration (IC50) of Rop was calculated by MTT assay;the SH-SY5Y cells were divided into control group (PBS for 48 h),Rop group (Rop at IC50 for 48 h),BDNF+Rop group (20 μg/L BDNF for 2 h,and Rop at IC50 for 48 h),Akt pathway activator SC79+Rop group (5 mg/L SC79 for 2 h,and Rop at IC50 for 48 h),and BDNF+Akt pathway inhibitor API-2+Rop group (20 μg/L BDNF+10 μmol/L API-2 for 2 h,Rop at IC50 for 48 h);the morphological changes of the cells were observed under microscope;MTT assay was used to detect the cell activity;flow cytometry was used to detect the cell apoptosis,and immunohistochemistry was used to detect the expressions of Akt and PCNA;the expressions of B lymphoma-2 (Bcl-2),Bcl-2-associated X (Bax) and cysteine protease-3 (Caspase-3) were detected by reverse transcription (RT)-PCR and Western blotting.Western blotting was used to detect the expressions of Akt and phosphatidylinositol 3-kinase (PI3K).Results (1) As compared with the 0 mmol/L Rop group,the 1,2,3,4,and 5 mmol/L Rop group had significantly decreased cell activity,significantly increased apoptosis rate,and statistically smaller number of Akt and PCNA positive cells (P＜0.05).(2) As compared with the control group,the Rop group had significantly decreased cell activity,statistically increased apoptosis rate,significantly smaller number of Akt and PCNA positive cells,significantly decreased Bcl-2 mRNA and protein expressions,significantly increased Bax and caspase-3 mRNA and protein expressions,and significantly decreased phosphorylated-(p-) Akt and p-PI3K protein expressions;as compared with the Rop group,the BDNF+Rop group and SC79+Rop group had significantly higher cell activity,significantly decreased apoptosis rate,significantly larger number of Akt and PCNA positive cells,significantly increased Bcl-2 mRNA and protein expressions,statistically decreased mRNA and protein expressions of Bax and Caspase-3,and significantly increased p-Akt and p-PI3K protein expressions (P＜0.05);as compared with the BDNF+Rop group and SC79+Rop group,the BDNF+API-2+Rop group had significantly lower cell activity,significantly increased apoptosis rate,significantly smaller number of Akt and PCNA positive ceils,significantly decreased Bcl-2 mRNA and protein expressions,statistically increased mRNA and protein expressions of Bax and Caspase-3,and significantly decreased p-Akt and p-PI3K protein expressions (P＜0.05).Conclusion BDNF can alleviate ropivacaine-induced neuronal injury by activating Akt signaling pathway,consequently modulating the proliferation and apoptosis of neurons.

Objective: To establish a liquid chromatography-tandem mass spectrometry (LC/MS/MS) method for the determination of microcystin-LR (MC-LR) in drinking water, investigate its removal efficiency during tap water advanced treatment process and analyze its degradation products in the tap water. Methods: Two parallel water samples were collected from each point of tap water advanced treatment process in September 2015, November 2015 and January 2016, respectively, and treated by mixing, filtration, concentration, elution, nitrogen blow and re-dissolvement. The samples were analyzed by LC/MS/MS to determine the MC-LR concentration and its removal efficiency during treatment process. The combination of actual water enrichment (including source water enrichment of 50 times and 1 500 times concentrated, finished water enrichment of 50 times and 2 500 times concentrated) and laboratory simulated water (including the mixture of MC-LR and liquid chlorine in the mass ratio of 1∶10, 1∶20, 1∶100 and 1∶1 000, respectively) were used to qualitative analyze the MC-LR degradation products by Orbitrap mass spectrometry. Results: The linearity of MC-LR ranged from 2 to 200 μg/L with the detection limit of 0.007 9 μg/L and the limit of quantification of 0.026 3 μg/L. The recovery rate of MC-LR from different contration in drinking water were from 94.88%-101.47%. The intra-day precision was 2.51%-7.93% and the intra-day precision was 3.24%-8.41%. The average concentration of MC-LR in source water was (0.631±0.262) μg/L, 94.0% of which can be removed by ozone exposure while the concentrate was (0.038±0.016) μg/L, biological pre-treatment and chlorination. The remaining can hardly be removed by sand filtration, ozone exposure, activated carbon, ultrafiltration and other processes. The MC-LR average concentration in the finished water maintained at about (0.036±0.016) μg/L. Degradation products including hydroxy-microcystin, methyl-hydroxy-microcystin, methyl-microcystin were identified in the laboratory simulated water of the mixture of MC-LR and liquid chlorine in the mass ratio of 1∶10. Conclusion The established MC-LR detection method can be well applied to the monitoring of MC-LR in drinking water due to its simple pre-treatment process and good methodological validation parameters. The degradation products of treatment processes was different.