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Objective:To explore the application value of situational bilingual teaching in standardized residency training of gynecology.Methods:The study enrolled 64 gynecological residents of Guangzhou Women and Children's Medical Center, and they were randomized into experimental group and control group, with 32 residents in each group. The control group was taught by the traditional teaching method, using self-compiled teaching materials of our hospital. The experimental group was taught by the situational bilingual teaching mode, and the teaching materials were the same as those of the control group. The training time of the two groups was two months. After teaching, the training effect was evaluated through examination and satisfaction survey. SPSS 22.0 software was used for t test and chi-square test. Results:After training, the scores of theoretical knowledge, operation skills and case analysis in the experimental group were significantly higher than those before training and after training in the control group ( P<0.05). The learning interest, autonomous learning ability, theoretical knowledge mastery, clinical thinking ability and communication ability of the experimental group were significantly better than those of the control group ( P<0.05). The number of residents who were very satisfied with the training was significantly higher than that of the control group, and that of residents who were basically satisfied with the training were less than that of the control group ( P<0.05). Conclusion:The application of situational bilingual teaching in the standardized residency training of gynecology has good teaching effects, which can significantly improve the theoretical performance, clinical operation ability and case analysis ability of the training residents, and also improve the satisfaction of the training residents.
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Objective To observe the differential expression level of CD4+CD25+Foxp3+regulatory T cells (Treg) in liver of arsenic-exposed rats, explore the regulatory mechanisms on immunological of hepatic injury induced by arsenic, and provide a basis for prevention and treatment of the disease. Methods Thirty-two healthy Wistar rats were selected and randomly divided into control,low,medium and high arsenic dose groups by weight,8 rats per group. Rats in control group were given oral gavage of deionized water, while the other groups were given oral gavage doses of 2.00 g/L sodium arsenite(NaAsO2) according to their body weight for 6 days every week, the concentrations were 1.25, 2.50 and 5.00 ml/kg. After 4 months, liver tissue samples of rats were collected, the content of arsenic in liver was detected by inductively coupled plasma mass spectrometry (ICP-MS);the expression of Treg cells in liver was detected by immunohistochemistry; enzyme-linked immunosorbent assay (ELISA) was applied to detect the levels of interloukin-10 (IL-10),transforming growth factor beta 1 (TGF-β1), IL-6, IL-17 and IL-2. Results Compared with the control group [28.57 (17.64 - 35.64)μg/g], the content of arsenic in liver in low,medium and high arsenic exposed groups[M(P25-P75):638.30(527.91-802.58),591.64(513.82-723.16),792.55 (695.93 - 1 074.41) μg/g] increased, the differences were statistically significant(P < 0.05). Compared with low arsenic group, the content of arsenic in liver in high arsenic group increased, the difference was statistically significant (P < 0.05). Numerical density on area (NA) of positive Treg cells in medium,high arsenic exposed groups [(2.25 ± 0.50),(4.00 ± 2.16)A/cm2]was higher than that of the control group[(0.60 ± 0.54)A/cm2,P<0.05];NA of positive Treg cells in high arsenic exposed group was higher than that of the low arsenic exposed group[(1.50 ± 0.58) A/cm2, P < 0.05]. The expressions of the IL-10 in low, medium and high arsenic exposed groups [(5.58 ± 1.70), (6.78 ± 1.09),(7.18 ± 0.53)μg/L]were higher than that of the control group[(2.32 ± 0.83) μg/L,P<0.05];compared with low arsenic group, the expression of IL-10 in high arsenic group increased (P < 0.05); compared with control group [(1.46 ± 0.65) μg/L], the expression of TGF-β1 in high arsenic exposed group increased[(9.06 ± 3.60)μg/L, P<0.05];compared with control group [(2.33 ± 0.66)μg/L], the expression of IL-6 in high arsenic exposed group increased [(5.03 ± 1.39) μg/L, P < 0.05], compared with low arsenic exposed group [(2.46 ± 1.71) μg/L], the expressions of IL-6 in high arsenic exposed group increased, the difference was statistically significant (P < 0.05);the expression of IL-17 among control, low, medium and high arsenic exposed groups[(4.87 ± 1.64),(7.50 ± 2.74), (6.21 ± 1.47),(7.23 ± 2.68)μg/L]were not statistically significant (F = 1.429, P > 0.05); compared with control group [(16.30 ± 3.98) μg/L], the expression of IL-2 in high arsenic exposed group decreased[(9.93 ± 2.65) μg/L, P <0.05]. The content of arsenic in liver was positively correlated with the expression of IL-10, TGF-β1, IL-17, IL-6 (rs=0.696,0.463,0.632,0.502,P<0.05),and negatively correlated with the expression of IL-2(rs=-0.522,P<0.05). Conclusion With increasing of arsenic exposure level, the content of arsenic in liver and the expression of CD4+CD25+Foxp3+Treg have increased,the cytokines are secreted abnormally,liver immunological micro environment is disordered,immune tolerance is formed,and immune clearance is inhibited,which may play an important role in the occur and development of immunological liver damage induced by arsenic in rat.
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Objective To investigate the infiltration of T helper 17 (Th17) and regulatory T cells (Treg) in the liver of rats exposed to arsenic,and to investigate the roles of Th17 and Treg in infiltration of liver injury induced by arsenic.Methods Thirty-two Wistar rats,half male and half female,were randomly divided into control,low,medium and high arsenic dose groups by body weight via the random number table method,8 rats per group.Rats in control group were given oral garage of deionized water,while other groups were given oral gavage doses of 2.00 g/L sodium arsenite (NaAsO2) according to their body weight for 6 days every week,the concentrations of NaAsO2 were 1.25,2.50 and 5.00 ml/kg,respectively.After 4 months,liver tissue samples of rats were collected,the content of arsenic in liver was determined by inductively coupled plasma mass spectrometry (ICP-MS);Hematoxylin-eosin staining (HE) method was used to observe the morphological changes of liver tissue in rats;the protein expressions of interleukin-17A (IL-17A,the imflammatory factor secreted by Th17 cells) and Forkhead Box P3 (Foxp3,the lineage-specific transcription factor of Treg cells) were measured with immunohistochemistry.Results ① Arsenic content in liver of low,medium,and high arsenic exposed groups [63.83 (52.79-80.26),59.16 (51.38-76.58),79.26 (69.59-107.44) μg/g] were higher than those of the control group [2.86 (1.76-3.56)μg/g,P < 0.05],and the high arsenic dose group was higher than the medium arsenic dose group (P < 0.05).② With increasing doses of arsenic,the numbers of inflammatory cells in the liver tissue of rats were increased,and the liver tissue of the high arsenic dose group showed vacuolar degeneration and pathological changes in some areas.③ Compared with the control group,low and medium arsenic dose groups (0.001 + 0.001,0.010 ± 0.020,0.030 ± 0.080),the expression of IL-17A protein in the liver in high arsenic dose group were significantly increased (0.220 ± 0.130,P < 0.05),the differences were statistically significant between groups (F =14.776,P <0.05).The expressions of Foxp3 protein in the liver in low,medium,and high arsenic dose groups were significantly higher (0.270 ± 0.050,0.330 ± 0.040,0.320 ± 0.070) than that in the control group (0.070 ± 0.020),the differences were statistically significant between groups (F =56.990,P < 0.05).④ There was a positive correlation between hepatic arsenic levels and protein levels of IL-17A and Foxp3 in liver (r =0.48,0.81,P < 0.05).Conclusion Arsenic exposure can increase the content of arsenic in liver tissue of rats,which causes the changes of infiltration of Th17 and Treg cells,leading to the change of immune status,suggesting that Thl7 and Treg cells play an important role in the development of arsenic-induced immune injury.
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Objective To explore the mechanism of cell apoptosis of immortalized human keratinocytes (HaCaT cells) and protein expression related to this process after long term exposure to sodium arsenite (NaAsO2,1.0 μmol/L).Methods Malignant transformation model was set up through long-term exposure of HaCaT cells to 1.0 μmol/L NaAsO2.Cell passage for 0,1,7,14,21,28 and 35 generations in the process of malignant transformation were collected for measurement of cell apoptosis rate by flow cytometry,and apoptosis related proteins by Western blotting,including activation of cysteine protease 3,8 (cleaved-caspase-3,8),C/EBP homologous protein (CHOP),B-cell leukemia/lymphoma 2 (Bcl-2),and Bcl-2 associated X protein (Bax).Results Along with the arsenite treatment,the apoptosis levels were significantly decreased (F =26.770,all P < 0.05),the apoptosis levels (0.307 ± 0.049,0.213 ± 0.055,0.163 ± 0.057,0.147 ± 0.035,0.053 ± 0.012) of the 7th,14th,21st,28th and 35thgenerations of cells after arsenite treatment were lower than that of control group of the 0th generation (0.393 ±0.021,all P < 0.05).Compared between generations,there were statistical differences of the protein expression levels of cleaved-caspase-3,Chop,Bax and Bcl-2 in arsenite group (cleaved-caspase-3:1.000 ± 0.000,1.030 ± 0.027,1.104 ± 0.069,1.016 ± 0.087,0.838 ± 0.075,0.753 ± 0.082,0.677 ± 0.073;Chop:1.000 ± 0.000,1.059 ± 0.018,0.934 ± 0.095,0.976 ± 0.216,0.793 ± 0.136,0.651 ± 0.042,0.564 ± 0.056;Bax:1.000 ± 0.000,1.069 ± 0.037,1.028 ± 0.042,0.954 ± 0.118,0.641 ± 0.135,0.531 ± 0.132,0.429 ± 0.085;Bcl-2:1.000 ± 0.000,1.072 ± 0.023,1.249 ± 0.134,1.334 ± 0.143,1.633 ± 0.221,1.507 ± 0.152,1.461 ± 0.145,F =7.730,7.355,27.802,12.438,all P < 0.05),compared with control group of the 0th generation (1.000 ± 0.000) and the same generation control group (1.000 ± 0.000),after the 21st generation,the differences were statistically significant (all P < 0.05),while there was no difference of the protein expression levels of cleaved-caspase-8 (F =0.832,P > 0.05).Conclusion In the process of malignant transformation,the apoptosis levels of HaCaT cells are inhibited after long term sodium arsenite exposure through mitochondria and endoplasmic reticulum (ER) stress signaling pathways.
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To study the effect of practice while watching (PWW) teaching method in cardiopul-monary resuscitation training of standardized training residents. 196 standardized training residents were selected, and PWW teaching method was used to carry out cardiopulmonary resuscitation training and as-sessment. The results were significantly higher than that before the training, and the satisfaction rate of the residents to the PWW teaching method was 100%.PWW teaching method could improve the effect of stan-dardized training resident doctors in cardiopulmonary resuscitation,and increase the interest in learning.
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Objective To investigate the differential expression levels and significance of regulatory T cell (Treg) and T helper 17 (Th17) related immunologic factors in peripheral blood of arsenic-exposed rats.Methods Thirty-two Wistar rats were numbered by weight,randomly divided into four groups [control,low (1.25 ml/kg),medium (2.50 ml/kg),and high (5.00 ml/kg)],8 rats per group.Rats in control group were given oral gavage of deionized water,and low,medium and high arsenic exposed groups were given oral gavage doses of 2.00 g/L sodium arsenite (NaAsO2) according to their body weight for 6 days every week.After 4 months,the urine and peripheral blood samples of rats were collected,urinary arsenic (uAs) was detected by inductively coupled plasma-mass spectrometry (ICP-MS),the results were shown in [median (minimum and maximum)],uAs was corrected by urinary creatinine (uCr),the unit was μg/g Cr;enzyme-linked immune-sorbent assay (ELISA) was applied to detect the levels of Treg,Th17,T lymphocytes activation related immunologic factors [interleukin-10 (IL-10),transforming growth factor beta1 (TGF-31),IL-17,IL-6,IL-2],the results were shown in (x) ± s.Results The uAs of the rats was compared between control,low,medium,and high arsenic exposed groups [7.50 (3.16-9.81),72.34 (62.34-106.63),209.15 (154.41-232.20),369.73 (289.50-516.55) μg/g Cr],the differences were statistically significant (F =337.55,P < 0.05).IL-10 [(85.03 ± 7.11),(93.96 ± 8.14),(97.48 ± 6.23),(93.47 ± 4.41) ng/L],TGF-β1 [(72.88 ± 2.96),(81.45 ± 8.15),(86.08 ± 7.55),(90.29 ± 5.35) ng/L],IL-17 [(18.15 ± 3.66),(25.54 ± 5.59),(31.48 ± 5.74),(37.25 ± 7.36) ng/L],IL-6 [(83.68 ± 8.48),(85.14 ± 7.11),(89.78 ± 5.36),(93.48 ± 5.77) ng/L],and IL-2 [(80.65 ± 6.90),(73.86 ± 8.00),(69.93 ± 7.77),(62.06 ± 9.82) ng/L] of the rats were compared between control,low,medium,and high arsenic exposed groups,the differences were statistically significant (F =5.094,11.054,16.249,3.474,5.119,all P < 0.05).There were positive correlations between uAs and TGF-β1,IL-17 concentration (r =0.723,0.605,all P < 0.01),while IL-2 showed a negative correlation (r =-0.484,P < 0.05).Concltsion Arsenic exposure could affect the secretion of Treg and Th17 related immunologic factors,so as to the imbalance of anti-inflammatory and pro-inflammatory,which may play a role in the formation and development of arsenic-related immune injury.
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Objectives To investigate the effects of sodium arsenite (NaAsO2) on the expression ot microRNA-191 (miR-191) and tissue inhibitor of metalloproteinase 3 (TIMP-3) in human normal hepatic cells (L-02 cells).Methods L-02 cells were exposed to different doses of NaAsO2 [0 (control group),5,25,50 and 75 μmol/L]for 24 h,or treated with 5 and 25 μmol/L NaAs02 for 0 (control group),12,24 and 48 h.The miR-191 inhibitor was used to suppress the expression of miR-191.qRT-PCR was performed to detect the expression level of miR-191 and TIMP-3 mRNA,and the protein level of TIMP-3 was analyzed by Western blotting.Results Dose-effect study:There were significant differences in the expressions of miR-191,TIMP-3 mRNA and protein between the 5 groups (F =85.674,20.952,123.393,all P < 0.05).The expressions of miR-191 in all groups (1.702 ± 0.124,2.077 ±0.234,2.145 ± 0.105,2.003 ± 0.077) were higher than that of control group (0.990 ± 0.035,all P < 0.05);the mRNA expressions of TIMP-3 in 25,50,75 μmol/L groups (0.848 ± 0.067,0.804 ± 0.081,0.813 ± 0.076) were all lower than that of control group (0.996 ± 0.007,all P < 0.05),but there was no significant difference in the mRNA expression of TIMP-3 between the 5 μmol/L group and control group (0.939 ± 0.133 vs 0.996 ± 0.007,P> 0.05),and the protein expressions of TIMP-3 in all groups (0.846 ± 0.093,0.611 ± 0.123,0.554 ± 0.098,0.529 ± 0.067) were lower than that of control group (1.006 ± 0.003,all P < 0.05).Time-effect study:there were significant differences in the expressions of miR-191,TIMP-3 mRNA and protein between the exposure groups of 5 and 25 μmol/L (For 5 μmol/L:F =86.355,16.404,22.898,all P < 0.05;For 25 μmol/L:F =104.321,20.123,52.321,all P < 0.05).The expressions of miR-191 in all exposure groups of 5 and 25 μmol/L (1.392 ± 0.152,1.691 ± 0.167,2.018 ± 0.130 and 1.456 ± 0.167,1.946 ± 0.178,2.259 ± 0.256) were higher than those of control groups (1.001 ± 0.014,1.008 ±0.027,all P < 0.05);the mRNA expressions of TIMP-3 in 48 h exposure group of 5 μmol/L and all exposure groups of 25 μmol/L (0.824 ± 0.093 and 0.897 ± 0.033,0.815 ± 0.089,0.709 ± 0.103) were lower than those of control groups (1.004 ± 0.018,0.997 ± 0.057,all P < 0.05),but there were no significant differences in the mRNA expressions of TIMP-3 between the 12,24 h exposure groups of 5 μmol/L and control group (0.952 ± 0.072,0.929 ± 0.121 vs1.004 ± 0.018,all P > 0.05);the protein expressions of TIMP-3 in all exposure groups of 5 and 25 μmol/L (0.857 ±0.068,0.832 ± 0.106,0.691 ± 0.112 and 0.785 ± 0.097,0.620 ± 0.066,0.453 ± 0.075) were lower than those of control groups (1.006 ± 0.045,1.004 ± 0.078,all P < 0.05).The treatment of miR-191 inhibitor:there were significant differences in the expressions of miR-191 and TIMP-3 protein between different groups (F =104.306,67.015,all P < 0.05).The elevated expression level of miR-191 induced by NaAsO2 was significantly suppressed after transfected with miR-191 inhibitor (0.314 ± 0.094 vs 2.051 ± 0.371,P < 0.05),which in turn up-regulated the protein expression of TIMP-3 (1.965 ± 0.277 vs 0.541 ± 0.183,P < 0.05).Conclusion The expression level of miR-191 is elevated in response to NaAsO2 exposure,and miR-191 has subsequently suppressed the expression of TIMP-3,a potential target of miR-191.
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Purpose To observe the effects of midkine ( MK) on human breast cancer cell line MDA-MB-231 angiogenesis in vitro, and to explore its mechanism. Method shRNA interference was performed to silence the expression of MK in MDA-MB-231 cells, and Western blot was used to identify the expression of MK and EPCR. After MK and EPCR knockdown, or treated with anti protease-activated receptor 1 (PAR1) antibody, the culture medium of MDA-MB-231 cells were collected and the conditioned medium were pre-pared. Human umbilical vein endothelial cells ( HUVECs) were cultured with conditioned medium, and the endothelial cells prolifera-tion was detected by CCK-8 assay, cell migration was detected by transwell method, vasculogenic activity was assessed by Matrigel-based tube formation assay. Results After knockdown of MK, the protein level of EPCR was decreased in MDA-MB-231 cells. Com-pared with control, knockdown of MK and EPCR decreased the proliferation, migration and angiogenesis ability of HUVECs significant-ly (P<0. 05), and the effect of EPCR knockdown group was stronger than MK knockdown group (P<0. 05). After treated with anti-PAR1 antibody, the proliferation, migration and angiogenesis ability of HUVECs were decreased compared with control and EPCR knockdown group (P<0. 05). Conclusion MK promotes human breast cancer cell line MDA-MB-231 angiogenesis through EPCR /PAR1 signaling pathway in vitro.
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Purpose To observe the effect of MK on proliferation, migration and angiogenesis of human umbilical vein endothelial HU-VECs cells and to explore the role of MK in tumor angiogenesis. Methods siRNA targeting MK was used to downregulate MK expres-sion in human breast cancer cell line MDA-MB-231. Then, the experiment was divided into three groups: the untreated group, the empty vector transfection group and MK gene interference group. CCK-8 assay was used to detect the endothelial cells proliferation, tr-answell method was for detection of endothelial cell migration numbers, and matrigel in vitro small tube formation assay was used to sur-vey the state of tube formation. Expression of MK in siRNA transfection was identified by RT-PCR and Western blot. Results The MK gene interference group showed lower cell proliferation activity, less number of migration of cells and tube formation than the other two groups (P<0. 05). Conclusion Low-expressed MK in human breast cancer cell line MDA-MB-231 can inhibit proliferation, mi-gration and angiogenesis of endothelial cells, which shows that MK may play an important role in tumor angiogenesis.
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Purpose To investigate the expression of VEGF, NRP1 and the cancer stem cell marker CD44 in gastric carcinoma and their correlations. Methods High throughput technology of tissue microarray and immunohistochemical SP method were employed to investigate the expression of VEGF, NRP1 and CD44 proteins in 72 gastric carcinoma specimens and 52 normal gastric tissues. Immu-nohistochemistry records were reviewed, their correlations and the clinicopathological characteristics were evaluated. Results ( 1 ) The positive rates of VEGF, NRP1 and CD44 proteins were 76. 4%, 66. 7%, and 83. 3% in gastric carcinoma, respectively, with sig-nificant differences from normal gastric tissues (P<0. 01). (2) The positive expression of VEGF and NRP1 were closely correlated with infiltration depth, lymph node metastasis and TNM stage (P<0. 05). The positive expression of CD44 was closely correlated with Lauren type, differentiation degree, lymph node metastasis and TNM stage (P<0. 05). (3) In addition, the expression of CD44 was significantly correlated with VEGF and NRP1 in gastric carcinoma (rs =0. 578, rs =0. 278, rs =0. 316, P<0. 05). Conclusions VEGF, NRP1 and the cancer stem cell marker CD44 are highly expressed in gastric cancer, which are closely related to the biological behaviors of gastric cancer. Synergistic effects are observed between VEGF and NRP1, and they are closely related to the expression of the cancer stem cell marker CD44, which remains to be further studied.
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Purpose To construct a lentiviral ventor-mediated RNA interference of human midkine ( MK) gene and to provide the ba-sis for further experiment in vivo and in vitro. Methods Four RNAi sequences targeting human MK gene were designed, and cloned into the lentiviral vector to construct lentiviral vectors:GV115-MK-1, GV115-MK-2, GV115-MK-3 and GV115-MK-4. After transfec-tion into competent E. Coli bacteria, the candidate clones were identified by PCR and DNA sequencing. The titer of lentiviruses was determined after 293 T cells were co-transfected with GV115-MK, pHelper 1. 0 and pHelper 2. 0. The four kinds of recombinant lenti-viruses were used to infect human breast cancer cell MDA-MB-231, and the expression levels of MK mRNA were detected by real-time PCR. Results PCR analysis and DNA sequencing confirmed that the four inserted MK RNAi sequences were corrected. Strong green fluorescence was observed in the 293 T cells under the fluorescent microscope after co-transfection. The titer of the concentrated virus was 6 × 108 , 5 × 108 , 5 × 108 and 6 × 108 TU/ml, respectively. The MK expression in MDA-MB-231 cells infected with lentiviral vec-tors GV115-MK-1 was decreased by 87. 2% (P<0. 05). Conclusion Lentiviral RNAi vector for MK gene is successfully construc-ted, and it could effectively inhibit the expression of MK gene in MDA-MB-231 cells in vitro.
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ObjectiveTo assess the psychosomatic health status of patients with facial steroiddependent dermatitis by using Pittsburgh sleep quality index (PSQI),self-rating anxiety scale (SAS) and selfrating depression scale(SDS).MethodsPsychosomatic health status was evaluated by using PSQI,SDS and SAS in 92 patients with facial steroid-dependent dermatitis and 50 healthy controls.ResultsThe scores on depression and anxiety in patients with steroid-dependent dermatitis were higher than those in the controls (63.11 ± 10.29 vs.36.78 ± 2.46,53.24 ± 9.98 vs.31.29 ± 9.45,both P < 0.01).There were significant differences between the two groups in the quality of sleep.The general and mean PSQI global scores on subjective daytime function,sleep disturbances,time needed to fall asleep and sleep duration were significantly higher in the patients than in the controls.ConclusionsPatients with steroid-dependent dermatitis have a poor sleep quality with the presence of anxiety and depresssion.Psychosocial interventions are warranted for the treatment of patients with steroid-dependent dermatitis.
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ObjectiveTo investigate the intervention of level of atorvastatin calcium tablets (lipitor) on soluble CD40 (CD40L) and high sensitivity C-reactive protein (hs-CRP) in patients with angina.Methods 65 patients with coronary heart disease were divided into stable angina (SA group,n= 30) and unstable angina (UA group,n= 35).The vein blood was collected in the hospitalized morning,2,4,6 weeks after treatment and serum levels of hs-CRP and CD40L were measured and compared between the two groups.ResultsThe levels of soluable CD40L [(20.52± 2.91)μg/L] and CRP [(7.96±1.69) mg/L] in UA group were higher than SA group [(7.96±-1.35) tg/L and ( 1.58 ± 0.91 ) mg/L] (t = 21.705、18.493,both P< 0.05) before treatment,and after treatment,their levels in the two groups were significantly lower than pre-treatment (P<0.01).ConclusionsCD40L and hs-CRP may involved in the pathophysiology of unstable angina process and can be used as an indicator reflecting vulnerable plaque.Lipitor might enhance stability of atherosclerotic plaque and prevent acute coronary events by reducing levels of CD40L and hs-CRP.
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Objective: To study the effects of physical and chemical factors on hairy root of Panax ginseng and its glycoside production. Methods: The cell growth index and its glycoside yield in different culture condition such as different plant hormones mixed, mediums and time were compared. Results: After hairy root of Panax ginseng was precultivated on the MS medium with IBA (0.5%) at first 72 hours. the culture was cultured on the MS medium. During 6 weeks, dry weight of hairy root increased by 98.74 times, and the content of glycoside synthesized by hairy root is 5.190% and Re+Rg1 is 0.3271%, respectively. Conclusion:We have achieved the best condition of hairy root of Panax ginseng.