OBJECTIVE:To establish a method for content determination of eucalyptol in Chimonanthus Zhejiangensis. METH-ODS:GC was conducted with Zebron ZB-WAX capillary column with temperature programmed;FID detector was used with the temperature of 250 ℃;the volume temperature was 220 ℃ and the carrier gas was nitrogen with high purity. RESULTS:Eucalyp-tol had a good linear relationship in the range of 0.017 69-1.415 mg/ml(r=0.999 1);the RSDs of precisions,stability and reproduc-ibility tests were no more than 1.87%and the average recovery was 99.72%(RSD=0.65%,n=6).There were big differences in dif-ferent areas of the same month and the maximum was more than one time (1.064%-0.450%);there were also big differences in the same area of different months(0.633%-1.064%). CONCLUSIONS:The method is simple,accurate,fast and reproducible and suitable for the quality control of Chimonanthus Zhejiangensis.

Objective To investigate the role and mechanism of signal transducer and activator of transcription 4 (STAT4) in homocysteine (Hcy)-induced proliferation,migration and senescence of human aorta smooth muscle cells (VSMCs).Methods After VSMCs were incubated with Hcy (0-0.50 mmol/L) for various time periods,a CCK-8 assay was performed to determine proliferation activity.Modified Boyden chamber system (Transwell apparatus) was used to examine migration ability.A SA-β-gal staining kit was used to determine senescence status.Western blotting was performed to examine the expressions of STAT4 and phosphorylated STAT4 (p-STAT4) proteins,and ELISA was performed to examine MCP-1 expression.Results Hcy (from 0.05 mmol/L) significantly increased the proliferation of VSMCs (P ＜ 0.05).Hcy (from 0.10 mmol/L) significantly increased the migration of VSMCs (P ＜ 0.05).Hcy promoted the senescence of VSMC in a concentration-dependent manner,and significantly increased VSMC senescence when the concentration of Hcy reached 0.10 mmol/L (P ＜ 0.05).Hcy (from 0.10 mmol/L) treatment significantly increased the expression of p-STAT4 (P ＜ 0.05) and MCP-1 (P ＜ 0.05).Conclusions Hcy increases the proliferation,migration and senescence of VSMCs,probably by the activation of STAT4 pathway.

Objective To investigate the effects of valproic acid ( VPA ) on SD rats with experimental autoimmune encephalomyelitis ( EAE) and its possible immunomodulatory mechanism .Meth-ods Fifty female Sprague-Dawley ( SD) rats were randomly divided into five groups by random digit table , including control group (n=10), EAE group(n=10), low dose VPA treated group (100 mg/kg, n=10), median dose VPA treated group (300 mg/kg, n=10) and high dose VPA treated group (600 mg/kg, n=10).The SD rat model of EAE was induced by immunizing with a guinea pigs′spinal cord homogenate (GPSCH).Normal saline and various doses of VPA were given to rats in according groups twice a day from day 0 to day 19 ( close to the peak stage of EAE ) .The severity of EAE was scored according to the signs and symptoms.Pathological changes were observed through Hematoxylin-Eosin staining, and then the degrees of inflammatory infiltration were evaluated .The numbers of activated neuroglia that expressed Iba-1 in cerebral and lumber cords were counted by immunohistochemistry .The expression of IFN-γ, IL-17 and IL-10 in cer-ebral and lumber cords were measured by ELISA .Results Compared with EAE group , rats in the low, me-dian and high dose VPA treated groups had lower incidence of EAE and prolonged latency , but only the me-dian dose treated group showed significant alleviation in clinical symptoms (P<0.05).Both the median and the high dose treated group showed decreased inflammatory cell infiltration in CNS (P<0.05).Immunohisto-chemistry results showed that the numbers of activated microglia were significantly inhibited in rats treated with median and high dose of VPA in comparison with those in EAE group (P<0.05).Results of ELISA demonstrated that the expression of IFN-γand IL-17 in both median and high dose VPA treated groups were significantly decreased compared with those in EAE group (P<0.05), but only the median dose treated group showed a remarkably increased expression of IL-10 (P<0.05).Conclusion VPA, especially medi-um dose of VPA ( 300 mg/kg ) , had neuroprotective effects on rats with EAE .The possible mechanism might be associated with the inhibited activation of microglia and the increased percentage of anti -inflammato-ry cytokines .

Objective To explore the therapeutic effect and possible mechanisms of Xuebijing injection for treatment of rabbits with acute pulmonary thromboembolism (APTE). Methods Thirty-two New Zealand rabbits were randomly divided into normal control group, model group, low molecular weight heparin (LMWH) group and Xuebijing group by random number table, 8 rabbits in each group. APTE model was reproduced by re-infusing autologous blood clots, and nothing was done for the normal control group. After the model was established, intravenous injection of Xuebijing 1 mL/kg from ear marginal vein was given to the rabbits in Xuebijing group, the LMWH 100 U/kg was subcutaneously injected in LMWH group, the same amount of normal saline was given intravenously to model group and normal control group, twice daily for 7 days in all the groups. The changes of plasma D-dimer and endothelin-1 (ET-1) levels were detected by enzyme-linked immunosorbent assay (ELISA) after modeling for 8 hours and 1, 3, 5, 7 days;after 7 days the animals were sacrificed and their lung tissues were removed for the examination of ultrastructural changes under an electron microscope. Results With the prolongation of time, the D-dimer level in the model group was gradually increased, and on the 7th day after modeling, the level was significantly higher than that of the normal control, LMWH and Xuebijing groups (g/L:3.98±1.14 vs. 0.43±0.12, 1.05±0.13, 1.04±0.15, all P<0.05). After modeling for 5 days, the ET-1 level reached its peak in the model group, and it was lowered on the 7th day after modeling, but still significantly higher than that in the normal control, LMWH and Xuebijing groups (g/L:63.8±20.9 vs. 20.0±1.4, 36.5±8.3, 34.2±6.3, all P<0.05). The D-dimer and ET-1 levels in the LMWH and Xuebijing groups were significantly higher than those of the normal control group at each time point, but still obviously lower than those of the model group (all P<0.05), and the differences between the two treatment groups were not statistically significant at each time point (all P>0.05). Electron microscopic examination of lung tissue ultra-structure showed:in the model group, there were the lung tissue basement membrane rupture, the organelle structure in vascular endothelial cells and alveolar epithelial cells typeⅠandⅡnot clear, disappearance of microvilli on cell surface, cytoplasm concentration, nuclear chromatin condensation, a lot of debris due to nuclear lysis, formation of apoptotic bodies after shedding, and mitochondrial structural damage. In LMWH group, the capillary endothelial cell swelling, a lot of necrotized blood cells oozing, blockage of lumen, the cytoplasm of typeⅡalveolar epithelial cell loose, edematous and multiple lamellar bodies vacuolized were seen. In Xuebijing group, capillary endothelial cell swelling, necrotized blood cells blocking the lumen and seeping, the link basically normal, the type Ⅱ alveolar epithelial cell edematous, mitigation of lamellar bodies emptying were found. Conclusions Xuebijing can significantly reduce the plasma D-dimer and ET-1 levels, adjust the balance of coagulation and fibrinolysis, simultaneously decrease the pulmonary vascular permeability obviously in rabbits with APTE, thus it possesses certain protective effect for endothelial function leading to amelioration of lung injury and reduction of lung pathological damage in rabbits with pulmonary embolism.

Objective To analyze the dynamic changes of perinatal defects in Lishui city from 2005 to 2014.Methods According to hospital-based data,87 294 fetuses and infants aged from 28 weeks of gestation to 7 days after birth were monitored for birth defects in Lishui city from 2005 to 2014.Results Among 87 294 perinatal infants,birth defects were found in 1 914 cases with a incidence of birth defects of 219.3/ten thousand,which was on the rise year by year except 2009.The incidence of birth defects was 235.1/ten thousand (737/31 352) in urban areas,which was significantly higher than that in rural areas (210.4/ten thousand,1 177/55 942) (x2 =5.705,P ＜ 0.05).The incidence of birth defects of boys and girls was 221.1/ten thousand (1 005/45 453) and 212.8/ten thousand (890/41 822),respectively (x2 =0.707,P ＞ 0.05).Among 1 914 cases of defects,657 were detected prenatally and 1 257 case were detected within 7 days after birth.Congenital heart disease was the most common birth defect,followed by polydactyly,cleft lip,microtia and syndactyly.Birth defect accounted for 32.74％ (278/849) of all perinatal deaths and the ratio was on the rise since 2010.Conclusion There is an upward trend in incidence of birth defects in Lishui city in the last decade;the incidence of birth defects in urban areas is significantly higher than that in rural areas;the incidence of congenital heart disease is high,and birth defects are the main cause of perinatal death.

Forty two resident general practitioners (GPs) attending obstetrics and gynecology course in Beijing Hospital were divided into trial group and control group with 21 in each group. The 2-hour study content uterine fibroids was taught by the same teacher. For control group the traditional classroom teaching mode was used, while for the trial group the WeChat-based flipped classroom model was used. The teaching effectiveness was evaluated by means of paper examination, critical examination and questionnaire survey. The average scores and the clinical reasoning scores of trial were significantly higher than those of control group [(90.5±3.4) vs. (82.5±4.2) points and (94.5±1.5) vs. (84.5±2.0) points; t=4.28, P<0.05 and t=5.01, P<0.05, respectively]. Compared with the control group, the trial group presented more learning interest, higher learning efficiency and self-learning ability; and also the ability of expression and communication was improved, the ability of teamwork and the ability of analyzing and solving problems was enhanced more markedly in trial group (all P<0.05). The result indicates that WeChat-based flipped classroom teaching model can improve the learning efficiency in general practice residency training.

Objective To explore the expression of tankyrase (TNKS) and its relationship with WNT/β-catenin signal?ing pathway in lung acinar adenocarcinoma. Methods Seventy-two samples of single subtype alveolar like lung adenocarci?noma (lung adenocarcinoma group) and 67 specimens of normal lung tissue adjacent to carcinoma (adjacent to carcinoma group) were collected. Immunohistochemical method was used to detect expressions of TNKS, beta-catenin (β-catenin) and c-myc protein. The correlation of each protein expression in lung adenocarcinoma tissues was analyzed. The differential ex?pression of TNKS was detected by Western blot assay in two groups. Results Tankyrase protein was mainly expressed in cy?toplasm. The expression ofβ-catenin protein was mainly in cytoplasm and nuclear of lung adenocarcinoma. The expression ofβ-catenin was mainly in cytoplasm, and a small amount was in nuclear of the adjacent group. The c-myc protein was ex?pressed mainly in the nucleus. The positive expression rates of TNKS,β-catenin and c-myc protein were significantly high?er in lung adenocarcinoma group than those of adjacent to carcinoma group (P<0.05). The expression ofβ-catenin in cyto?plasm and nucleus was positively correlated with the expression of TNKS and c-myc (P<0.05). Western blot analysis showed that the relative expression level of TNKS was significantly higher in lung adenocarcinoma group than that of adja?cent to carcinoma group (0.497 ± 0.021 vs. 0.237 ± 0.015, t=13.00, P<0.01). Conclusion Abnormally high expression of TNKS in lung adenocarcinoma may promote the occurrence of lung cancer by regulating the WNT signaling pathways. Inhib?iting TNKS expression may become a new target to treat lung adenocarcinoma.

Background Primary conjunctival lymphoid hyperplasia exhibit atypical clinical symptoms and easy to cause misdiagnosis and mistreatment.Understanding the clinical characteristics of primary conjunctival lymphoid hyperplasia is of an important clinical significance.Objective This study was to analyze the clinical characteristics and therapeutic outcome of patients with primary conjunctival lymphoid hyperplasia.Methods The medical records of 20 patients with primary conjunctival lymphoid hyperplasia who was treated in Beijing Tongren Hospital from January 2012 to December 2014 were retrospectively analyed.The age,symptoms,therapy,prognosis,histopathological features and immonochemistry results were reviewed,and the treating approach targeting to different types of conjunctival lymphoid hyperplasia was evaluated.The fellow-up duration was 1-2 years.Results Age of the patients ranged from 27 to 83 years old.The lesions were classified as benign lymphocytic hyperplasia,atypical lymphoid hyperplasia and lymphoma.Majority of lesions located in fornix conjunctiva tissue (90％).Follicle-like appearance was seen in the benign lymphocytic hyperplasia,and pink elavation was exhibited at the bulbar conjunctiva in the atypical lymphoid hyperplasia.A diffuse salmon fish like appearance in conjunctiva was the primary feature of conjunctival lymphoma.Histopathological examination showed that conjunctival benign lymphocytic hyperplasia had follicle-like tissue in lesions,atypical lymphoid hyperplasia exhibited diffuse lymphocyte filtration,and conjunctival lymphoma appeared monocyte filtration.The 3 tpyes of lesions also could be differentiated by immunochemistry.The medicine was applied in the eyes with benign lymphocytic hyperplasia,and the combination of resection of lesions with conjunctival tissue or amniotic membrane transplantation was used for the atypical lymphoid hyperplasia or lymphoma eyes.No recurrence of the lesions was found during fellow-up duration.Conclusions The clinical manifestations of conjunctival lymphoid hyperplasia lesions are varied,and it is easy to be confused with chronic ocular surface diseases.Pathology and immunochemistry are helpful for the differential diagnosis.The treating regimen is dependent on the lesion type.Most patients have a favorable prognosis with treatment.

Aim To explore the therapeutical effect and mechanism of baicalein on two 6-hydroxydopamine (6-OHDA ) induced Parkinson′s disease (PD ) rat models,which received unilateral lesions of the left medial forebrain bundle (MFB ) or caudate putamen (CPu ) made by stereotaxic injection of 6-OHDA (MFB-M,CPu-M).Methods PD rat models were established by microinjection of 6-OHDA into MFB or CPu.The anti-tremor effect of baicalein on PD rat models was examined.Spontaneous activity was recor-ded. Dopamine (DA ), dihydroxyphenylacetic acid (DOPAC)and homovanilic acid (HVA)in striatum were quantified by HPLC-ECD.The tyrosine hydroxy-lase (TH)and OX-42 positive cells were detected by immunohistochemical method.The morphological vari-ation of the neurons was confirmed by analysis at an ul-trastructural level.Results Baicalein significantly in-creased the spontaneous activity in CPu-M.The elec-tromyography (EMG ) recordings revealed that com-pared with 6-OHDA group,the tremor frequency in ba-icalein group was decreased by 55% in MFB-M,and by 60% in CPu-M.6-OHDA treatment decreased DA levels in the striatum,while treatment with baicalein attenuated the DA decreases in CPu-M.Moreover,ba-icalein treatment could increase TH-positive neurons and decrease OX-42-postive microglia compared with 6-OHDA group in both MFB-M and CPu-M.Conclu-sions In the present study,it is illustrated that ①microinjection of 6-OHDA into the MFB and the CPu could cause different pathological changes of PD, which is important for efficacy evaluation;②baicalein showed the ability to alleviate the behavior symptoms in PD-rats at different stages by improving motor function and attenuating muscle tremor;③therapeutic effect of baicalein was produced by inhibiting the inflammatory medium production and release,anti-apoptosis,chan-ging dopamine catabolism, and inhibiting dopamine turnover.

OBJECTIVE:To improve the quality standard of Jianpi zhixiening granules. METHODS:TLC was applied for qual-itative identification of Scutellaria baicalensis,Crategi Fructus,Lonicerae japonicae,Codonopsis Radix,Nelumbo nucifera,Copti-dis Rhizoma. The contents of berberine hydrochloride and baicalin were determined by HPLC. The determination was performed on Shimadzu VP-ODS column with mobile phase consisted of methanol-acetonitrile-water (46:30:42,V/V/V,berberine hydrochlo-ride))(0.1% sodium dodecylsulphate,0.1% phosphoric acid,methanol-0.4% phosphoric acid(50:50,V/V,baicalin)at the flow rate of 1.0 mL/min. The detection wavelengths were set at 265 nm (berberine hydrochloride) and 280 nm (baicalin). The column temperature was 30 ℃,and sample size was 10 μL. RESULTS:TLC spots of S. baicalensis,Crategi Fructus,L. Japonicae,Co-donopsis Radix,N. nucifera,Coptidis Rhizoma were clear and well-separated without negative interference. The linear ranges of berberine hydrochloride and baicalin were 6.67-33.34 μg/mL(r=0.9998)and 7.7-38.7 μg/mL(r=0.9999). RSDs of precision,sta-bility and reproducibility tests were all lower than 1.0% . The recoveries were 96.5% -99.9%(RSD=1.2% ,n=6)and 101.1%-102.9%(RSD=0.6%,n=6). CONCLUSIONS:Improved standard can be used for quality control of Jianpi zhixiening granules.