Objective To investigate the effects of ursolic acid on the expression of extracellular signal regulated kinase(ERK1),C-Jun,C-Myc,Cyclin D1 in transplanted tumor of malignant glioblastoma cell line C6 in nude mice and the related mechanisms.Methods C6 glioblastoma cells were subcutaneously injected into nude mice to establish the subcutaneous model of glioblastoma in nude mice,and then the mouse models were divided into 3 groups: blank control group,ursolic acid group(50 mg?kg-1?d-1,intra-abdominal injection,20 d),PD98059 group(2 mg?kg-1?d-1,intra-abdominal injection,7 d).Survival of nude mice,growth and histopathological changes of the tumors were observed.The expressions of ERK1,C-Jun,C-Myc,and Cyclin D1 in the tumor tissues and the expression of ERK1 mRNA in the tumor tissues were detected by immunohistochemical technique(IHC) and in suit hybridization(ISH),respectively.Results Slow growth of the implanted tumor was found in ursolic acid group and PD98059 group.The mean volume and weight of tumors in the two groups were significantly smaller than those in the blank control group(P

Objective:To investigate the distribution of calcitonin gene-related peptide immunoreactive(CGRP-IR) nerve fibers in periodontal tissue of rats, providing morphological data of CGRP in periodontitis. Methods:Five adult SD rats were sacrificed by transheart perfution fixation with 4% paraformaldehyde. The mandibles were removed and decalcified. Frozen sections were made and CGRP-IR nerve fibers in periodontal tissue were observed with immunohistochemical technique. Results: CGRP-IR nerve fibers were observed in gingiva, alveolar bone .periodontal membrane; many heavily stained CGRP-IR nerve fibers were found beneath junctional epithelium, epithelium of gingival sulcus and epithelium of gingival col; some CGRP-IR nerve fibers even penetrated into junctional epithelium and epithelium of gingival col. Conclusion; CGRP-IR nerve fibers extensively disdtribute in the periodontal tissue with the densest in the initial area of periodontitis, suggesting that CGRP may play a role in the genesis and development of periodontitis.

Objective: To evaluate the impact of fine particulate matter ( PM2.5 ) on respiratory diseases and symptoms of community residents in Chun’an County, so as to provide the basis for air pollution treatment strategies. Methods: Using the cluster random sampling method, the permanent residents in Qiandaohu Town of Chun’an County were investigated from 2017 to 2018. The demographic information, respiratory diseases and symptoms were collected by using Surveillance Plan for the Impact of Air Pollution ( Haze ) on health ( 2016 Edition ). The air pollutants and meteorological data were collected through Qiandaohu station of Hangzhou Ecology and Environment Monitoring Center and Hangzhou Meteorological Information Center. The effects of PM2.5 on respiratory diseases and symptoms of residents were analyzed with generalized estimating equation. Results: Totally 1 181 people aged 6 months to 95 years were recruited, including 557 ( 47.16% ) males and 624 ( 52.84% ) females. Acute nasopharyngitis (common cold) and tracheitis/tonsillitis occurred most frequently, with 203 cases, accounting for 3.44%. The median of daily average concentration of PM2.5 was 24 μg/m3, with the standard exceeding rate of 2.80%. The results showed that PM2.5 increased the risk of acute nasopharyngitis ( common cold ), tracheitis/tonsillitis, cough, expectoration, runny nose, sore throat and nasal congestion ( lag 3 days, OR: 1.015-1.022, 95%CI: 1.001-1.037 ); the effect of PM2.5+PM10 ( OR: 1.020-1.040, 95%CI: 1.006-1.070 ) and PM2.5+O3 ( OR: 1.017-1.024, 95%CI: 1.005-1.035 ) was greater than that of PM2.5 alone on respiratory diseases and symptoms. Conclusion Atmospheric PM2.5 exposure in Chun’an County increases the risk of respiratory diseases and symptoms among community residents.

Objective To compare the curative effect of perindopril and metoprolol in the treatment of diabetic patients with asymptomatic cardiac insufficiency.Methods According to the method of random numbers, 152 diabetic patients with asymptomatic cardiac insufficiency were divided into the observation group and control group,with 76 cases in each group.The observation group was given with perindopril treatment,while control group was given with metoprolol treatment.The cardiac function index,glycosylated hemoglobin,BNP levels,the incidence of cardiac insufficiency and the occurrence of adverse drug reactions before and after treatment were compared.Results There were no significant differences in LVEF,Tei index and E/A between two groups before treatment (all P >0.05 ).Compared with before treatment,the cardiac function indexes of the two groups were significantly improved after treatment(t=5.96,4.83,4.91,10.22,5.68,5.18,all P<0.05),but the observation group were improved more significantly(t=5.64,4.67,5.03,all P<0.05).Before and after treatment,there was no significant difference in the levels of glycosylated hemoglobin(all P>0.05),and the difference between the two groups was not statistically significant(all P>0.05).The BNP levels of the observation group before and after the treatment were (515.43 ± 58.85)pg/mL and (214.43 ±32.42)pg/mL respectively,of which the control group were (513.94 ±59.11)pg/mL and (263.32 ±43.31)pg/mL respectively,and after treatment,the BNP levels of the two groups were significantly decreased(t=23.45,17.68,all P<0.05),of which the observation group decreased more obvious(t=6.97,P<0.05).The cardiac insufficiency incidence of the observation group was 3.95%,which was significantly lower than 14.47% of the control group(χ2 =5.03,P<0.05).The incidence of adverse drug reactions had no obvious differ-ences in both groups(χ2 =0.08,P>0.05).Conclusion Using perindopril in treatment of diabetic patients with asymptomatic heart function can obviously improve cardiac function,reduce the incidence of BNP and cardiac insuffi-ciency.It has the clinical curative effect and high safety.

Many Chinese medicinal materials are difficult to be distinguished in characters resulting in easy confusion. In the pa-per, the basic concepts of three groups of easily confused Chinese medicinal materials ( schisandrae chinensis fructus and schisandrae sphenantherae fructus, phellodendri cortex and phellodendri amurensis cortex, akebiae caulis and clematidis armandii caulis) were illus-trated and the development of verification methods were reviewed in the respect of modern instrumental analysis technology. With the improvement of analysis technology, the identification of Chinese medicinal materials relies on experimental data instead of artificial ex-perience, which makes it more objective and accurate.

Objective：To investigate the changes of calcitonin gene-related peptide immunoreactive (CGRP-IR)nerve fibers in rat dental pulps during acute inflammation. Methods： Rat acute pulpitis model was established by silk thread ligation and the change of CGRP-IR nerve fibers was observed with immunohistochemical method.Results： In radical pulp,the CGRP-IR nerve fibers became denser and more heavily stained；in the coronal pulp,the number of CGRP-IR nerve fibers decreased,but the background staining was heavier. Conclusion： During acute inflammation,the amount of CGRP increases in dental pulps, and is released into the surronding tissue in a large scale in the coronal region.

Objective To reseach the time point of the highest percentage of neural precursor cells derived from adipose stromal cells (ADSCs) in vitro, and to observe the ultrastructure features of neural precursor cells. Methods Used the β-mercaptoethanol to induce ADSCs to differentiate into neural precursor cells and neuron-like cells. The morphology of the uninductedcells and inducted cells were observed with inverted phase contrast microscope. The expression of nestin which was the marker of neural precursor cell in each group was detected using immunofluorescence staining method. The ultrastructural feature of cells which was induced for 3 hours were observed. Results The highest ratio of positive expression of nestin was 3 hours following induction,with the ratio ( 86.25 ± 4.82) %. There were many protuberance on the cell membrane under transmission electron microscopy.There were plenty of organelles in the neural precursor cells. The neural precursor cells had a large size nucleus,large nucleoplasmic index, much extended chromatin,and less condensed chromatin. The nucleus had double-layer nuclear envelope, more nuclear pore on the nuclear envelope. Conclusion The time point of the highest percentage of neural precursor cells derived from ADSCs is 3 hours,and the ultrastructral feature of induced neural precursor cells confirm that cells at this time point are in a state of split active period.

This study aimed at optimizing the extraction process of Bi Xie Fen Qing (BXFQ) drink using ultrasound-microwave cooperation with central composite design-response surface method in comparison with the traditional decoction process.Taking microwave frequency,amount of water,extraction time as the main detection factors,diosgenin,glycyrrhizic acid and the extract yield were tested as the evaluation indexes;and taking an integrated score as response value for response surface and contour,predictive analysis was carried out and the optimum extraction conditions were achieved.It was found that the optimum extraction process of BXFQ drink was identified:the microwave frequency was 434 W,water addition was 18.4 times,extraction time was 9.3 mins and the ultrasonic frequency was fixed at 50 W.Under the optimum process,the diosgenin extraction yield rate was 23.17％ (mg· g-1),extraction yield rate of glycyrrhizic acid was 0.64％ (g·g-1),and the extraction yield rate of extractum was 34.12％ (g·g-1).All the indexes were superior to those of the traditional method.It is concluded that the composite design-response surface method is suitable for the extraction optimization of BXFQ drink with favorable predictability of the mathematical model.The optimized ultrasoundmicrowave cooperation was easy to operate with high extraction efficiency.It is suitable for industrialized production with the provision of a scientific reference for the modern formulation development of BXFQ drink.

Objective To induce adult adult adipose-derived stem cells(ADSCs) in vitro to differentiate into neuronal-like cells,and to analyze the features of their cell morphology and ultrastructure. Methods Adipose stromal cells were obtained and amplified in vitro. Then make use of chemical induction to induce them. Observed ADSC and differentiation of cells in morphology and ultrastructure under inverted microscope and transmission electron microscopy. Immunocytochemistry to detection of Nestin, Neuron Specific Endolase( NSE) ,and glial fibrillary acidic protein(GFAP) expression in cells. Used Real-time PCR to detection of Nestin,GFAP gene mRNA expression before and after induction in ADSC. To observe the morphology and ultrastructure of the cells prior to and after induction under microscope and electron microscope. Results The morphology of ADSC was similar to fibroblasts ,and could be amplified stability within 10 passages in vitro. Some of the cells induced display a typical astrocyte-like cells in ultrastructure. Followed neuronal induction,astrocyte-like cells began to stain brightly for CFAP, Nestin. GFAP stained in both the cytoplasm and nuclei of astrocyte-like cells, but Nestin only stained in the cytoplasm. The peak positive expression rate within 14d following neuronal induction. The rate of positive expression cells was( 14.4 ± 3. 6) % for Nestin, (87. 3 ± 5. 3 ) % for GFAP. Then two kinds of protein expression remained the similar rate. The average relative concentration of GFAP and Nestin gene mRNA have significant statistical difference between ADSC and differented cells analyzed by Real-time PCR (P<0.05).The peak concentration of GFAP was within 20 d after induction,and GFAP was within 14 d after induction. Conclusion In the cytoplasm of adult adipose-derived cells possess Nestin genetic material,which is the marker of neural stem cell. The differential astrocyte-like cells have the typical morphology, ultrastructure and GFAP phenotype of mature astrocytes.

Objective To observe nitric oxide synthase (NOS) activity and the expresssion of inducible NOS (iNOS) mRNA in cultured calf aortic endothelial cells treated with various concentrations of insulin. Methods Cultured calf aortic endothelial cells were incubated with different concentrations of insulin for 24 h. NOS activity was determined by colorimetry and iNOS mRNA was measured by semi-quantitative RT-PCR. Results NOS activity and the expression of mRNA in calf aortic endothelial cells at pharmacological concentration (10 -7 mol/L) of insulin were significantly increased than those at physiological concentration (10 -10 mol/L) of insulin (P