Objective To explore the effect of progesterone on the expression of O4 and O1 in the white matter of neonatal rat model with periventricular leukomalacia (PVL). Methods 2-day-old neonatal SD rats were randomly divided into model group, experimental group, and sham operation group. Rats' left common carotid artery was ligated and exposed to hypoxia (8%O2+92%N2) for 0.5 h in both the model group and experimental group to build the PVL animal model. The rats in experimental group was injected intraperitoneally with progesterone 10 mg/(kg·d) immediately after cerebral hypoxia ischemia. In sham operation group, rats' left common carotid artery was only isolated without ligation and hypoxia. 1, 4, 7, and 14 days after operation, the pathological changes of brain tissue were compared among three groups. Immunohistochemical staining was used to detect the expression of O1 and O4 in the cerebral cortex of rats in three groups at different time points. Results There were no abnormal pathological changes in the white matter in the sham operation group at each time point. The left ventricular enlargement and periventricular leukomalacia were found in both model and experimental groups, while the pathological damages of white matter in experimental group were significantly lighter than those in model group at each time point. The integral optical density (IOD) of O1 and O4 positive cells in the cerebral cortex of the three groups was gradually increased at day 1, day 4, and day 7 after operation and reached the peak level at day 7 , then was decreased at day 14 after operation. There was statistically significant difference (P<0.01). At day 1, day 4, day 7, and day 14, the integral optical density (IOD) of O1 and O4 positive cells in the cerebral cortex of sham operation group was highest, followed by experimental group and model group, and there was significant difference (P<0.01). Conclusion Progesterone can reduce the pathological damage in the cerebral cortex in neonatal rats with PVL, and promote the expression of O1 and O4 in the periventricular white matter, which can promote the differentiation and maturation of oligodendrocytes.

Objective To determine the expression of CD4+ CD25+ FOXP3+ regulatory T cells(Treg cells) in peripheral blood of patients with hepatocellular carcinoma (HCC) and investigate the clinical significance of Treg cells determination in clinical practice. Methods Flow cytometry was employed to measure the levels of CD4+ CD25+ T cells and CD4+ CD25+ FOXP3+ T cells in peripheral blood of 18 HCC patients, 26 hospitalized patients without HCC (clinical control) as well as 24 healthy persons (healthy control). Results The percentage of CD4+ CD25+ T cells in total CD4+T cells isolated from the HCC patients(4.25% ± 3.98 % ) was elevated significantly compared to that in the clinical control group (1.34% ± 1.14%) or healthy control group (1.29% ±0.95%) (both P＜0.01). There was no difference in the percentage between the clinical control group and the healthy control group (P＞0.05). Meanwhile, the ratio between CD4+ CD25+ FOXP3+ T cells and CD4+ T cells in HCC patients (2.94%0.91%) also increased significantly compared to that in the clinical control group (0.76% ± 0.34%) or healthy control group (0.81% ± 0.29%) ( both P＜0. 001), which showed a more obvious increasing tendency than the ratio of CD4+ CD25+ T cells and CD4+ T cells. No difference in percentage of CD4+ CD25+ FOXP3+ T cells in CD4+ T cells was found between the clinical control group and the healthy control group (P＞0.05). Conclusion As the more accurate regulatory T cells, CD4+ CD25+ FOXP3+ T cells are able to detect the increase of population in HCC patients. Therefore, it is important to determine the levels of CD4+ CD25+FOXP3+ T cells in HCC patients for prevention and treatment of malignancy.

Objective To observe the in vitro inhibitory effects of brucea javanica oil on human washed platelet and explore the possible mechanism.Methods Human washed platelets were mixed withdifferent concentration of brucea javanica oil which were divided into four groups[untreated control group,negative control group,9.0% of brucea javanica oil group,and 22.5%of brueea javanica oil group].The maximal ratio of platelet aggregation induced by adenosine liphosphate(ADP),arachidonic acid(AA),collagen,and thrombin,respectively,was measured with platelet aggregation analyzer.The expressions of fibrinogen receptor(FIB-R)and P-selectin(CD_(62p))on external membrane of activated platelet were determinated with flow cytometry.The F-actin of cytoskeletal structure in activated platelet was detected by SDS-PAGE.Results At 9.0% of brucea javanica oil,the maximal ratio of platelet aggregation[(57.7±4.0)%,(62.2±3.9)%,(66.9±5.0)%and(71.8±5.1)%]induced by ADP,AA,collagen,and thrombin,was significantly lower than that[(75.3±4.1)%,(79.3±4.8)%,(80.6±5.4)%,(84.1±6.2)%]at negative control(0% of brucea javanica oil)(P<0.01),but makedly higher than that[(39.2±3.5)%,(45.8±3.4)%,(51.2±3.9)%and(56.7±4.8%)]at 22.5%,respectively(P<0.01).The inhibitory rate of platelet aggregation(47.9%,42.2%,36.5%and 32.6%)at 22.5%of brucea javanica oil was notably higher than that(23.4%,21.6%,17.0%and 14.6%)at 9.0%,respectively(P<0.001).There was a negative correlation between brucea javanica oil concentration and the aggregation ratio of platelet stimulated by the four agonists,respectively(r=-0.952,-0.961,-0.970,-0.975,P<0.001).At 9.0% of brucea javanica oil,the expression levels of FIB-R[(64.7±4.0)%]and CD_(62p) [(3.91±0.21)%] of platelet activated by ADP were significantly lower than that[(85.5±4.6)%and (5.05±0.27)%]at negative control,but remarkably higher than that[(36.2±3.9)%and(2.34±0.15)%]at 22.5%,respectively(P<0.01).There was a much higher inhibitory rate of platelet aggregation(57.7%)at 22.5%than that(24.3%)at 9.0%(P<0.01).The ratios(1.68±0.10 and 1.77±0.12)of F-actin photodensity at 22.5%and 9.0%to that in blank control were significantly lower than that(2.22±0.15)at negative control(P<0.01)but there was no statistical difference between the ratios in the group of 9.0%and 22.5%brucea javanica(P>0.05).Conclusions brucea javanica oil has special inhibitory effect on activated platelet and thrombosis in a dose-dependent manner.The mechanism is to inhibit the expression of fibrinogen receptor on external membrane of activated platelet,which is also related to the inhibition of F-actin and secretion of platelet.

To investigate the chemical constituents of Physalis minima L. , compounds were isolated by chromatographic methods from Physalis minima L. . Their structures were determined on the basis of spectral analysis. Five compounds were isolated and identified as Withaphysalin P(I), 14, 18-di-O-acetylwithaphysalin C(II), Withaphysalin Q(III), Withaphysalin 1(IV)and Withaphysalin 2(V). Compounds IV and V are new compounds, orderly named as Withaphysalin T and Withaphysalin U.

Objective To construct stable expression system of pLVX-Pro-CCR7 and study the function of CCR7 gene in metastasis of stomach cancer. Methods PCR method was used to amplify CCR7 gene and construct pLVX-Puro-CCR7 recombinant plasmid. Lipofection was used to infect SGC7901 cell strain immediately, and puromycin was used to screen the cell strain and establish a cell strain with stable expression. Western blot method was used to test the ex-pression of CCR7 in SGC7901 cells which were stably infected. Transwell method was used to analyze the effect of overly expressed CCR7 gene on in vitro migration of stomach cancer cells of SGC7901. Living imaging was used to ob-serve the metastasis ability of stomach cancer cell strain of SGC7901 which overly expressed CCR7 in nude mice. Re-sults pLVX-Puro-CCR7 recombinant plasmid was successfully constructed and SGC791 cell strain was stably infected. The cell strain was able to correctly translate and express CCR7 protein. Metastasis ability in nude mice and in vitro migration ability of SGC7901 stomach cancer cell strain which overly expressed CCR7 gene significantly improved (P<0.05). Conclusion CCR7 may be involved in the function of promoting metastasis of stomach cancer cells.

Objective To analyze the influence factors for complete embolization of intracranial aneurysms. Methods The clinical data of 546 inpatients with single intracranial aneurysm underwent interventional embolization at the Department of Neurosurgery,Anhui Provincial Hospital Affiliated to Anhui Medical University from January 2013 to January 2017 were analyzed retrospectively. They were divided into either a complete embolization group (n=255) or a incomplete embolization group (n=291) according to the immediate embolism degree of aneurysms. Single factor,multiple factors logistic regression analyses were used to analyze the factors associated with complete embolization of intracranial aneurysms. Results Univariate analysis showed that there were significant differences in the rupture status,anatomical morphology,Hunt-Hess grade, aneurysm size and neck width, different treatment regimens, and aneurysm angle between the patients in the complete embolism group and the incomplete embolism group ( all P<0. 05). The results of multivariate regression analysis showed that aneurysm size ( OR,0. 344,95%CI 0. 204-0. 578,P<0. 01),aneurysm rupture status (OR,0. 568,95%CI 0. 314-0. 947,P=0. 030), embolism ways (OR,3. 699,95%CI 2. 223-6. 153,P<0. 01),neck width of aneurysm (OR,0. 326, 95%CI 0. 198-0. 539,P=0. 003),aneurysm angle (OR,0. 647,95%CI 0. 451-0. 928,P=0. 018),and aneurysm morphology (OR,1. 689,95%CI 1. 118-2. 552,P =0. 013) were the independent factors of affecting the complete embolization of intracranial aneurysms. Conclusion Tiny, unruptured, narrow-neck, small inclination angle,regular-shaped aneurysms,stent-assisted or balloon-assisted embolization of intracranial aneurysms are easier to embolize the aneurysms completely.

Objective To establish and assess the new method of APTT assay based on the combinations of Mg2+and Ca2+for lupus anticoagulants(LA)measurements.Methods This prospective study included 309 trisodium citrate anticoagulated plasma samples from 244 random patients and 65 patients with different autoimmune diseases(AID)to establish and assess the method of LA measurement, respectively.Final concentrations of 0,2.0, 4.0, 8.0,16.0 mmol/L Mg2+were added into 25 mmol/L Ca2+solution, and Actin reagent was used to measured plasma APTT of 94 patients.The applied concentration of Mg2+-Ca2+solution was confirmed through the special and significant alteration of APTT from LA-positive and -negative plasma observed in the presence of Mg 2+(test solution).Based on Actin reagent use,the test solution and 25 mmol/L Ca2+solution were applied to measure APTT of patients and normal individuals, respectively, and the ratio of Mg2+-Ca2+-APTT to Ca2+-APTT(Mg2+-Ca2+-APTT indices)and normalized Mg2+-Ca2+-APTT indices(NAR)were calculated, respectively.Mixed plasma NAR was measured,and CV%was calculated to evaluate the repeatability and stability of Mg 2+-Ca2+-APTT method.APTT of 150 patients was measured with the test solution and Actin reagent to calculate Mg 2+-Ca2+-APTT indices, and normalized LA ratio was determined with dRVVT method.The applicability of Mg2+-Ca2+-APTT assay was assessed through comparisons of the results from the two methods.Finally, NAR and NLR of 65 patients with AID(including 26 SLE patients)were measured with Mg2+-Ca2+-APTT assay and dRVVT method, respectively, and ROC curve was also used to assess the efficacy of the two methods for LA measurements.Results In all LA-negative plasma,APTT increased from 28.1 ±4.5 s to 61.2 ±7.9 s in normal APTT group,47.2 ±8.9 s to 97.5 ±10.3 s in increased APTT group,and 27.6 ± 5.1 s to 61.2 ±7.9 s in ACA-positive group when Mg2+increased from 0 to 8 mmol/L in Mg2+-Ca2+solution(F=34.12, 38.9 and 28.35,P<0.01).Following increased Mg2+concentration, APTT shortened from 0 to 4.0 mmol/L, but simultaneously prolonged from 4.0 to 16.0 mmol/L in LA-positive plasma with prolonged or normal APTT(F=31.55 and 39.51, P<0.01), and APTT was significantly higher in 8.0 mmol/L than that in 4.0 mmol/L(P<0.001).The test concentration of Mg 2+/Ca2+solution was 4.0 mmol/L.The within, inter-day CV% of NAR was 1.39%,2.30%, and 3.44%, respectively. According to the judging criteria of <0.966 and >1.034 of Mg2+/Ca2+indices, there was 141 patients with increased indices and NLR <1.20, and 9 patients with decreased ones and NLR≥1.20 in all 150 patients.The area under ROC curve of NAR and NLR for LA detection was 0.913(95%CI:0.848-0.978) and 0.892(95%CI:0.817-0.966), respectively, and the cut-off value was 0.87 and 1.13, respectively. The sensitivity and specificity of NAR(85% and 77%)was higher than that of NLR(81% and 74%), respectively.The accordant rate of positive,negative,and total results between NAR and NLR was 94.4%, 98.5%,and 98%,respectively.Conclusion The method of APTT assay based on Mg2+combining Ca2+for LA measurements is feasible,and can be used to detect plasma LA of patients.