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Objective:To investigate the clinical value of peripheral blood T lymphocytes in the diagnosis and prognosis of patients with hepatocellular carcinoma (HCC) recurrence after liver transplantation.Methods:The clinical and laboratory data of 50 HCC patients, who received liver transplantation and were followed up in the Liver transplantation Center of Beijing Youan Hospital from January 2014 to December 2016, were retrospectively analyzed. The differences on clinical laboratory indicators and five-year survival were compared between HCC recurrence group ( n=29) and non-recurrence group ( n=21). Spearman correlate analysis was used to analyze the correlation between clinical laboratory indicators and HCC recurrence after liver transplantation. Receiver operator characteristic (ROC) curve was used to analyze the diagnostic value of CD4+T lymphocytes in HCC recurrence after liver transplantation. Kaplan-Meier survival curve was used to compare the survival time of patients with different CD4+T lymphocytes levels post liver transplantation. Results:Compared to non-recurrence group, the level of alanine aminotransferase, aspartate aminotransferase, γ-glutamyltransferase, albumin, lymphocytes, alpha-fetoprotein, protein induced by vitamin K deficiency or antagonist-Ⅱ, CD3+, CD4+and CD8+T lymphocytes were significantly different (all P<0.05). The median recurrence time after liver transplantation was 13.0 (6.0, 24.0) months, and the mortality rate was 100%. The 5-year mortality rate was 0 in the non-recurrence group. During 5-year follow-up, the median survival time of patients in the HCC recurrence group was 18.0 (9.0, 36.0) months, which was significantly lower than that of non-recurrence group [60.0 (60.0, 60.0) months, ( P<0.05)]. Compared with non-recurrence group, the CD3+, CD4+, and CD8+T lymphocytes were significantly lower in the recurrence group (all P<0.05). Spearman correlate analysis showed that HCC recurrence after liver transplantation was negatively correlated with the CD3+, CD8+and CD4+T lymphocytes ( r=-0.43, -0.38, -0.44, all P<0.05). ROC analysis showed that CD4+T lymphocytes at cutoff of≤265.50 cells/μl was valuable for the diagnosis of HCC recurrence after liver transplantation (specificity 100%, sensitivity 48.30%). Survival curve analysis showed that the survival time was significantly lower in the CD4≤265.50 cells/μl group [15.0 (10.0, 36.8) months] than that in the CD4>265.50 cells/μl group [53.0 (19.5, 60.0) months] ( P<0.05). Conclusion:There is a significant negative correlation between CD4+T lymphocytes and HCC recurrence after liver transplantation. CD4+T lymphocytes at cutoff value of≤265.50 cells/μl is valuable for the clinical diagnosis and prognosis evaluation of HCC recurrence after liver transplantation.
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Objective:To explore the clinical and laboratory characteristics of acquired immunodeficiency syndrome (AIDS) patients complicated with disseminated nontuberculous mycobacterial (NTM) disease, in order to provide basis for clinical therapy.Methods:The clinical findings, imaging and etiological data of the 71 AIDS patients complicated with disseminated NTM disease admitted to Beijing You′an Hospital from June 2016 to June 2021 were retrospectively analyzed.Results:Among 71 patients with disseminated NTM disease, the most common initial symptom was fever, followed with cough, expectoration, fatigue, poor appetite, abdominal pain and diarrhea. Seventy point four percent (50/71) of these patients had at least two comorbidities, with oral candida infection, cytomegalovirus infection, syphilis, pneumocystis pneumonia and bacterial pneumonia being the most common. Hemoglobin ((87.8±24.2) g/L) and albumin ((27.3±7.0) g/L) levels significantly decreased, while erythrocyte sedimentation rate ((59.8±28.6) mm/1 h) and C-reactive protein ((74.7±50.8) mg/L) levels increased in most cases. The median CD4 + T lymphocyte count was 7×10 6/L. The median time of positive blood culture of NTM was 260 h. Among the 71 patients, 40 cases (56.3%) were infected with Mycobacterium avium, 15 cases (21.1%) with Mycobacterium intracellulare, 10 cases (14.1%) with Mycobacterium colombiense, three cases (4.2%) with Mycobacterium marseillense and three cases (4.2%) with Mycobacterium kansasii.The frequent imaging findings were patchy and nodular shadows in lungs, and most patients had mediastinal or hilar lymph node enlargement and splenomegaly. Conclusions:AIDS complicated with disseminated NTM disease is prevalently occurred in patients with severe immune deficiency, and most of the bacteria belong to the Mycobacterium avium- intracellulare complex. Early obtaining positive etiological results of NTM is essential to guide the correct clinical diagnosis and accurate treatment.
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Objective@#To understand the characteristics and clinical significance of anti-soluble liver antigen antibody (anti-SLA) in patients with liver diseases.@*Methods@#Serum samples from seventy-seven patients with anti-SLA were collected from Beijing You'An Hospital during the period between January 2010 and December 2018. Anti-SLA, anti-liver cytosol type 1 antibody (anti-LC1), anti-glycoprotein 210 antibody(anti-gp210) and anti-nuclear body protein sp100 antibody(anti-sp100) were detected by immunoblotting; indirect immunofluorescence assay used for detecting anti-nuclear antibody (ANA), anti-mitochondrial antibody (AMA), anti-smooth muscle antibody (SMA), and anti-liver kidney microsome antibody (anti-LKM). One-way analysis of variance was used to compare the ages of different anti-SLA groups. The non-parametric rank sum test was used to compare the liver function indexes and immunoglobulins in different intensity groups of anti-SLA. P<0.05 was considered statistically significant. Further comparisons were made between the two groups, the correction level α′=0.008 3, P<0.008 3 was considered statistically significant.@*Results@#The average age of 77 anti-SLA positive patients was (52.50±1.25) years old, 70 females (90.9%) and 7 males (9.1%). 80.5% of anti-SLA-positive patients (62/77 cases) were strongly positive at the time of initial diagnosis (+++ to ++++).The Alanine aminotransferase (ALT) level in the SLA++ group was higher than that in the SLA++++ group (232.7 U/L vs 65.6 U/L,χ2=7.751,P=0.005) and the immunoglobulin M(IgM) level in the SLA+++ group was lower than that in the SLA++++ group (1 270 mg/L vs 2 270 mg/L,χ2=8.337,P=0.004).There was no significant difference in age, other liver function and immunological indicators among the different groups.Seventy cases (90.9%) were both anti-SLA and ANA positive, 13 cases (16.9%) were positive with SMA, and none positive with anti-LKM and anti-LC1. Among anti-SLA positive patients, 58 cases were diagnosed with autoimmune hepatitis (AIH), 12 were AIH/primary biliary cholangitis (PBC) overlap syndrome (OS), 2 were drug-induced liver injury, 2 were chronic hepatitis B, and 3 were hepatitis A, hepatitis E and acquired immune deficiency syndrome (AIDS) with liver injury, respectively.Cases of AIH and AIH/PBC OS accounted for 90.9% (70/77 cases) of anti-SLA-positive patients, and 5 of 7 patients diagnosed with non-AIH (and OS) had elevated IgG, showing AIH feature.92.3% (12/13 cases) of anti-SLA with high titers of AMA were diagnosed as AIH/PBC overlap syndrome. Of the 77 anti-SLA-positive patients, 28 (36.4%) had advanced or end-stage liver disease, including decompensated cirrhosis (22 cases), chronic acute liver failure (4 cases), and liver transplantation (1 case) and death from liver failure (1 case).@*Conclusions@#Anti-SLA has high diagnostic specificity for AIH;anti-SLA positive in patients with PBC should be an important biomarker for the diagnosis of AIH/PBC overlap syndrome.
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Objective@#To analyze the serological characteristics of anti-mitochondrial antibody M2 subtype (AMA-M2) in patients with drug-induced liver injury (DILI) and primary biliary cholangitis (PBC), in order to provide reference for clinical differential diagnosis.@*Methods@#Laboratory data of 2802 DILI cases who visited the hospital between January 2011 and December 2017 were retrospectively collected. AMA-M2 positive patients were analyzed with respect to laboratorical findings, and serum data of 120 patients with primary biliary cholangitis (PBC) at the same period was taken as a control. A chi-square test was used for group comparisons. One-way ANOVA and rank sum tests was used for ALT, AST, ALP, GGT and three groups of immunoglobulin M.@*Results@#Among 2802 DILI patients, AMA-M2 positive rate was 5.1% (144/2 802), 77.1% (111/144) was DILI alone, 22.2% (32/144) was DILI with PBC, and 0.7% (1/144) was DILI with Sjogren's syndrome. An AMA-M2 level in DILI alone group was mostly mild and moderate than the PBC group and the DILI combined with the PBC group. There was significant difference between the two groups (P < 0.05).There was no significant difference in AMA-M2 levels between DILI group combined with PBC group and PBC group (P > 0.05). ALT and AST levels of DILI alone group and DILI combined with PBC were (585.92 ± 653.04) U/L, (501.45 ± 512.67) U/L and (373.47 ± 502.60) U/L, (335.97 ± 513.96) U/L, respectively, which were significantly higher than PBC group [(106.33 + 134.08) U/L, (112.59 + 152.20) U/L]. There were statistically significant differences between the two groups (P < 0.05).The ALP level of DILI alone group was (152.58 + 81.46) U/L, which was lower than PBC group (237.86 + 215.09). The difference was statistically significant (P < 0.05). The level of immunoglobulin M in the DILI alone group was (1.76 ± 1.16) g/L, which was lower than PBC group (4.74 ± 5.74) g/L and the DILI combined with the PBC group (3.31 ± 1.68) g/L. There was significant difference between the two groups. During follow-up, 2.7% of patients with DILI had cirrhosis, 42.3% had lower AMA-M2 titer, 14.4% had lower AMA-M2 titer, 13.5% had higher AMA-M2 titer and five cases developed PBC.@*Conclusion@#AMA-M2 is not only positive in patients with PBC, but also low-to-medium or even high-level AMA-M2 may be detected in DILI patients. For AMA-M2-positive DILI patients, it is necessary to identify whether they are associated with PBC. Secondly, the levels of ALT, AST and ALP should be analyzed, and the patients should be on regular follow up for early and timely detection of drug-induced PBC.
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Objective To compare the test performance of different immunoassays for the detection on autoantibodies specific to primary biliary cholangitis,including anti-mitochondrial type 2 antibody(AMA-M2),anti-glycoprotein 210(anti-gp210)and anti-nuclear body protein sp100(anti-sp100).Methods Serum samples from Primary Biliary Cholangitis(PBC, n=91), liver disease control(including viral hepatitis,autoimmune hepatitis and liver cirrhosis,n=67)and healthy individual(n=40)were collected from Beijing Youan Hospital during the period between April 2014 and April 2017.All samples were tested with chemiluminescent immunoassay(CLIA)and enzyme linked immunosorbent assay(ELISA)for AMA-M2, meanwhile the detection on anti-gp210 and anti-sp100 were compared between CLIA and Line Immunoassay(LIA).The Kappa coefficient were used to measure the level of qualitative agreement between different assays.The diagnostic accuracy of AMA-M2 detected with CLIA and ELISA were compared by receiver operating characteristic curve(ROC).Results The overall qualitative agreement between CLIA and ELISA for the detection to AMA-M2 is 88.4%(Kappa =0.765, P<0.01).Excellent qualitative agreement between CLIA and LIA for the detection to anti-gp210 and anti-sp100 was also found with overall agreement as 96.5%(Kappa=0.852,P<0.01)and 98%(Kappa=0.884,P<0.01), respectively.The ROC analysis also showed similar area under the curve(AUC)for CLIA(0.965, P<0.01)and ELISA (0.928,P<0.01)on detection to AMA-M2.Conclusions CLIA and ELISA showed excellent agreement for the detection to AMA-M2.High qualitative agreement between CLIA and LIA was also found when testing anti-gp210 and anti-sp100.
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Objective To investigate the diagnostic value of serum FER, AFP and AFP-L3 alone or in combination for diagnosis of primary hepatic carcinoma( PHC).Methods This was a case-control study. Serum FER, AFP and AFP-L3 were determined in 212 patients with PHC ( StageⅠ45 cases, StageⅡ78 cases, StageⅢ81 cases, StageⅣ8 cases) , 127 patients with cirrhosis, 101 patients with chronic hepatitis and 98 controls in the Beijing Youan Hospital affiliated to Capital Medical University from January 2014 to December 2014.Levels of FER, AFP and AFP-L3 were measured by chemiluminescence, while serum samples were pre-treatment with affinity adsorption before AFP-L3 detection.FER, AFP and AFP-L3 levels were analyzed using the nonparametric Wilcoxon test among all groups.Diagnostic performance were analyzed among the groups with the three biomarkers independently and combined.Logistic regression, plotted ROC curve and calculated the area under ROC curve ( AUC) were applied to assess the diagnostic value of each index.Results Serum concentration of FER in PHC, cirrhosis, chronic hepatitis groups and healthy controls were 308.45 ( 148.98 -662.80 ) , 151.70 ( 51.44 -507.40 ) , 298.20 ( 157.30 -701.80 ) , 113.50( 54.98-221.38) μg/L, respectively.The concentration of AFP were 48.50(5.25 -748.40), 3.91(1.80-17.53), 4.76 (2.29-30.56), 2.57 (0.93-3.68) μg/L in each group.The serum levels of AFP-L3 in each group were 4.75(0.61-127.95), 0.61 (0.61-2.50), 0.61 (0.61-2.85), 0.61 (0.61-0.61) μg/L.The concentration of FER, AFP and AFP-L3 differs statistically in PHC, cirrhosis, chronic hepatitis group and healthy controls (χ2 =67.66,146.31,119.02,P<0.001).The content of serum FER, AFP and AFP-L3 increased gradually as the stage level aggravating ( StageⅠ-Ⅳ) , there was significant differences among groups (χ2 =21.63,22.68,21.98, P<0.001) .When using one serum marker, FER had the highest sensitivity (75.00%) , while AFP-L3 had the highest specificity (82.52%). While using two serum markers, FER/AFP had the highest sensitivity (89.15%) , FER+AFP-L3 and AFP+AFP-L3 had a higher specificity (86.20%).The combined detection of FER/AFP/AFP-L3 improved the sensitivity of the test to 89.15%, while FER+AFP+AFP-L3 had a specificity of 86.50%.The AUC of combination of FER, AFP and AFP-L3 was 0.803 ±0.019 (95% CI:0.765-0.841), which was higher than the AUC of either FER(0.748 ±0.022,95% CI:0.705-0.790, Z=4.67,P<0.001) and AFP-L3 (0.726 ±0.024,95% CI: 0.679 -0.772, Z=3.64,P<0.001).However, there was no significant difference in AUC between AFP alone ( 0.776 ±0.021, 95% CI: 0.735 -0.818 ) and the combined detection ( Z=1.34, P=0.18 ) .Conclusions FER was a potential marker for PHC diagnosis.The combination of FER, AFP and AFP-L3 has higher value of clinical applications than one of them independently.
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Objective To determine the efficacy of berberine in the treatment of non-alcoholic steatohepatitis ( NASH) , and to investigate the regulating effect on macrophage phenotype transformation in hepatic tissue on methionine -choline deficiency (MCD) diet induced NASH mice.Methods Fourty male C57BL/6 mice were randomly divided into 4 groups (10 mice per group): the normal group (fed with normal diet), the NASH model group (fed with MCD diet), rosiglitazone treatment group (30mg/kg) and berberine treatment group (150mg/kg).Drugs were adopted in the preventive intervention method for 2 weeks.The hepatic histopathological method was adopted to evaluate the drug therapeutic effect.The serum levels of tumor necrosis factor-α(TNF-α), interleukin(IL)-6, and IL-10 were examined with ELISA method.M1 and M2 phenotype were detected by flow cytometry .Results The results showed berberine improved the degree of hepatic histopathology .Berberine not only reduced the level of TNF-α, but also increased the level of IL-10 in serum on NASH mice significantly ( P <0.05 ) . Flow cytometry data indicated that berberine decreased M 1 type macrophages and increased M 2 type macrophages in liver tissue .The ratio of M1/M2 was significantly decreased in berberine and rosiglitazone treated group ( P <0.01 ) .Conclusion Berberine may improve the hepatic pathological process in MCD diet induced NASH model possibly through modulating macrophage phenotype transformation , i.e.The ratio of M2 type is more than M1 type in hepatic tissue , and increasing anti-inflammatory cytokines .
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<p><b>OBJECTIVE</b>To investigate liver fibrosis-related changes of CD4⁺CD25⁺Foxp3+ regulatory T cells (Tregs) in peripheral blood and in liver-infiltrating lymphocytes (LILs) using a mouse model.</p><p><b>METHODS</b>C57BL/6 mice were randomly divided into a model group and a control group. The model group received intraperitoneal injection of carbon tetrachloride (CC1₄) to induce liver fibrosis, and the control group received an equal volume of physiological saline. Serum was collected for detection of alanine aminotransferase level. Histopathological changes in liver were assessed by microscopic observation of tissues stained by hematoxylineosin and Masson. Frequencies of peripheral and intrahepatic Tregs, NK1.1⁺ cells, CD4⁺ and CD8⁺ cells were analyzed by flow cytometry. Intrahepatic Foxp3+ cells were detected by immunofluorescence. Liver expression of IL-6, IL-10, TGFb and Foxp3 was measured by RT-PCR detection of mRNA .Inter-group differences were evaluated by t-test.</p><p><b>RESULTS</b>The model group showed a significantly higher frequency of intrahepatic CD25⁺Foxp3⁺/CD4⁺ Tregs (10.63% ± 1.50% vs. control group:1.80% ± 0.66%; P less than 0.01) but only slightly higher frequency of peripheral Tregs (6.00% ± 0.62% vs. 5.33% ± 2.86%). The model group also showed significantly higher levels of intrahepatic Foxp3+ cells and of Foxp3 mRNA (both P less than 0.05), but significantly lower frequencies of NK1.1 cells in LILs (9.53% ± 2.25% vs. 19.80% ± 5.97%; P less than 0.05) and in peripheral blood (0.38% ± 0.13% vs. 1.06% ± 0.63%; P less than 0.05). The CD4⁺ cell frequency and the CD4⁺/CD8⁺ ratio were lower in LILs and peripheral blood of the model group, but none differed significantly from the control group. The intrahepatic expression of TGFb mRNA was significantly higher in the model group (P less than 0.01).</p><p><b>CONCLUSION</b>In liver fibrosis, intrahepatic CD4⁺CD25⁺Foxp3+ Tregs are increased while NK1.1+ cells are decreased. Tregs may suppress NK1.1+ cells through a mechanism involving TGFb.</p>
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Animals , Male , Mice , Liver , Metabolism , Pathology , Liver Cirrhosis, Experimental , Blood , Metabolism , Pathology , Mice, Inbred C57BL , T-Lymphocytes, RegulatoryABSTRACT
Objective To investigate the effect of blockading OX40-OX40L co-stimulatory signaling on the survival time of liver allograft in rat.Methods siRNA-expression vectors were constructed to targeting OX40.3～5 minutes before DA to Lewis orthotopic liver transplantation was performed,5×109 pfu of targeting OX40 siRNA plasmid DNA were diluted in 5 ml of phosphate buffered saline(PBS)and inlected intravenously into recipient Lewis rat over a period of 10 seconds.Serum IL-2 and IFN-γ levels were assayed by ELISA,and mix lymphocyte response(MLR)were tested by 3H-thymidine.Results The survival time of recipients in siRNA treatment group(74.0±9.3)was significantly longer than that in control group[(7.3±0.5)days].In experiment group,the inflammatory cell infihration and liver tissue structure destruction were very slight.The concentration of serum IL-2 was much lower in siRNA treatment group[(46±8.4)pg/ml]than that in control group[(286.5±14.6)pg/ml].Meanwhile,the concentration of serum IFN-γ was much lower in siRNA treatment group [(202.7±14.6)pg/ml]than that in control group[(1682.7±87.9)pg/ml].Conclusion Administration of OX40-siRNA can blockade OX40-OX40L co-stimulatory signaling pathway.hence inhibit the rejection of liver allograft.
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OBJECTIVE: To observe the effects of Tribulus terrestris L. saponion (TTLS) on apoptosis in cortical neurons induced by hypoxia-reoxygenation in rats. METHODS: Primary culture of rat cortical neurons was performed in vitro. A model of apoptosis of cortical neurons was established by hypoxia and reoxygenation. Hypoxia for 3 h in neural cells was induced with mixture of 95% N(2) and 5% CO(2), and then reoxygenation in neural cells was induced with mixture of 95% O(2) and 5% CO(2) for 12 h. Different concentrations of TTLS were administered to traditional Chinese herbal medicine-treated group separately during hypoxia and reoxygenation. The apoptosis rate was analyzed quantitatively by flow cytometry with Annexin V-FITC and propidium iodide staining. Mitochondria membrane potential was observed by a confocal laser-scanning microscope with JC-1 fluorescence. Caspase-3/7 activity in cytoplasm was measured by fluorescent plate reader. Bax protein expression was observed by immunohistochemical technique. RESULTS: The percentage of apoptosis was significantly increased, mitochondria membrane potential was obviously decreased, fluorescence of caspase-3/7 activity was increased, and Bax protein was abundantly expressed followed by 3 h of hypoxia and 12 h of reoxygenation (P<0.01). TTLS could inhibit the depression of membrane potential induced by hypoxia and reoxygenation, decrease the activity of caspase-3/7, reduce the expression of Bax protein, and inhibit the apoptosis of the cortical neurons. CONCLUSION: Hypoxia and reoxygenation can induce apoptosis of rat cortical neurons. TTLS can decrease the apoptosis induced by hypoxia and reoxygenation. The mechanism might be related to stabilization of mitochondria membrane potential, inhibition of caspase activity and reduction of Bax protein expression.
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AIM: To investigate the anti - tumor effect of Weimaining (WMN) on a murine Lewis lung carcinoma (3LL) and the influence on the cell cycle. METHODS: The inhibitory rate of WMN in 3LL growth was detected by replicating the model of 3LL. The effect of the drug on 3LL cell cycle and the influence of the drug on the expression of cy clin D1 protein were investigated by flow cytometry and immunohistochemical staining. RESULTS: The results showed that the inhibitory rate of drug in 3LL is 19. 14%, 33.59%, 40. 63% and 51.56% respectively at dosage ranging from 100,cells in G0 -G1 phase and decreases the expression of cyclin D1 protein. CONCLUSION: WMN inhibits the growth of 3LL cells in vivo by decreasing the expression of cyclin D1, blocking the cells in G0 - G1 phase and preventing the cells transition from G1 to S phase while DNA is replicated.
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AIM: To investigate the effects of Chinese medicine, Jinan injection, on ultrastructure and mitochondria in cultured lung cancer cell lines. METHODS: The cultured lung cancer cell lines PG and PAa were used and divided into 4 groups: control (C), cisplatin (DDP), Jinan (JA) and Jinan in combination with cisplatin (DJ), respectively. The changes of morphology and mitochondria membrane potential, intracellular Ca~(2+) and pH in every group were observed by inverted microscope and electronic microscope as well as by using flow cytometry, staining by rhodamine, Fluo-3 and BCECF, respectively. RESULTS: Degeneration cells showed chromatin condensation and peripheral congregation. In cytoplasm autophage lysosome increased and myelinoid body was seen easily. In mitochondria structure, where the space between the inner and outer membranes of these organelles expanded as the matrix was compressed. The electron-dense or swelled was observed as vacuole degeneration and its matrix showed electron-lucent. Compared to control, mitochondria membrane potential increased in every group after 24 h and 48 h treatment. DDP increased intracellular calcium ion in PG cells, however, in PAa cells, JA and DJ decreased it. Intracellular pH got lower at 24 h and higher at 48 h in PG and PAa cells. There were significance in every group vs control in PG and PAa by statistic t-test (P
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AIM: To investigate the effect of Qingjiening(QJN) on cytokines in sheep red blood cell(SRBC)-immunized mice. METHODS: After immunization of mice with SRBC, the effect of QJN o n IL-1、IFN-?、IL-2 in mice was observed, the IFN-? level was measured by macrophage NO - 2-release assay, the IL-1 level was measured by thymocyte a ssay, the IL-2 level was measured by mitogen activated lymphocytoblast assay. RESULTS: QJN can significantly inhibit mice to secrete IL-1、IFN- ? and IL-2. CONCLUSION: The immunosuppressive activity of QJN may be associate d with inhibition of immunocyte to secret IL-1、IFN-? and IL-2.
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AIM: To study the effect of Tribulus terrestris L. saponin (TTLS) on apoptosis and changes in cytosolic calcium concentration induced by hypoxia/re-oxygenation in rat cortical neurons. METHODS: Rat cortical neurons in primary culture were used, and a apoptosis model was induced by hypoxia/reoxygenation. LDH releasing rate was detected by spectrophotometry. The apoptosis rate of cortical neurons was analyzed quantitatively by flow cytometry with Annexin V-FITC and PI staining. Intracellular free Ca2+([Ca2+]i) was observed with a confocal laser-scanning microscope and determined by mean fluorescent value with Fluo-3 fluometry. RESULTS: Compared to control group, three hours of hypoxia and twelve hours of reoxygenation group induced cortical neuronal apoptosis and significantly increased the intracellular free Ca2+ concentration(P
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AIM: To observe the dynamic alteration of myristoylated alanine-rich C kinase substrate(MARCKS) mRNA expression in rat hippocampus with acute multi-cerebral infarction,and discuss the relationship between the alteration of hippocampus MARCKS gene and ischemia damage.METHODS: The acute multi-cerebral infarction model was established by method of Kaneko.Neurological function deficits were evaluated in the behavior test.The consequences of cerebral ischemic damage were examined by histopathological analyses.The MARCKS mRNA expression was measured by semi-quantitative PCR.RESULTS: The rats in acute multi-cerebral infarction group showed different level changes of neurological function deficits.The hippocampus damage of histopathology became significant 24h after ischemia.At the same time,the MARCKS mRNA expression was upregulated at the area of rats hippocampus during ischemia,and its overexpression started 1h after ischemia,and reached maximum7d after ischemia.CONCLUSION: MARCKS mRNA of rat hippocampus overexpresses during acute cerebral ischemia.This MARCKS mRNA overexpression is related with hippocampus ischemia damage.
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AIM: To observe the effect of puerarin on myocardial injury according to the time of occurrence of myocardial injury in the development of type 2 diabetic mice.METHODS: The serum levels of glucose(GLU),triglyceride(TG),total cholesterin(TC),low density lipoprotein-cholesterol(LDL-C) and high density lipoprotein-cholesterol(HDL-C) in 17-week,20-week,24-week,28-week KKAy mice were detected by automatic biochemical methods.The apoptotic percentage of cardiomyocytes was examined by flow cytometry.The expressions of bax and bcl-2 mRNA in cardiomyocytes were detected by RT-PCR.Caspase-3 expression in cardiomyocytes was determined by immunohistochemical staining.RESULTS: Compared to normal control mice,not only GLU level increased,but also the levels of TG,TC,LDL-C,HDL-C in 20-week,24-week and 28-week KKAy mice increased apparently(P
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AIM: To observe the effect of Shenmai injection,a chinese medicine,on apoptosis of cardiac myocytes after hypoxia.METHODS: Cardiac myocytes were separated from neonate rat heart and cultivated in vitro.Hypoxia condition was induced by mixture of 95%N2 and 5%CO2.Cells were exposed to hypoxia for 6 h or 12 h and treated with Shenmai injection(5 mL/L) from 24 h before hypoxia until the end of hypoxia.First,apoptosis was detected with Annexin V-FITC and PI staining by flowcytometry.Then,the activity of cardiac myocyte mitochondria was observed by MTT method.Mitochondria membrane potential and the activity of caspase 3,7 were also measured by laser scan microscopy and multi-detection microplate reader,respectively.RESULTS: The apoptotic cells became more and more with prolonged hypoxia.Shenmai injection enhanced mitochondria activity,kept membrane potential,inhibited the activation of caspase3,7 and then decreased apoptotic cells(P
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AIM: To investigate the anti-metastasis effect of weimaining, extracted from fragopyrum cymosum meissn, a Chinese medicine, on murine Lewis lung carcinoma (3LL). METHODS: The anti-metastasis effect of weimaining in vivo was detected in the grafting lung metastasis model of murine Lewis lung carcinoma. The effects of the drug on the expression of CD34 and E-cadherin were investigated by immunohistochemical staining and RT-PCR. RESULTS: Weimaining effectively inhibited the lung metastasis of 3LL at a concentration of 250 mg?kg -1?d -1, significantly suppressed the expression of CD34 and increased the expression levels of E-cadherin protein and mRNA in 3LL cells. CONCLUSIONS: Weimaining inhibits the metastasis of murine Lewis lung carcinoma (3LL) in vivo via increasing the expression of E-cadherin and decreasing microvessel density of tumor tissue.
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Objective:Based on the model of human umbilical vein endothelial cell(HUVEC) treated by tumor necrosis factor-alfa(TNF),We investigate the effects of muscone on polymophonulear leukocytes(PMN)-HUVEC adhesion and its adhesion molecules(CAMs) expression.Methods:Confocal system was used for identifying cultured HUVEC,MTT assay for its activity,Rose Bengal Staining for PMN-HUVEC adhesion,and fluorescent-immunocytochemistry techniques for CAMs expression.Results:After HUVEC treated by TNF,the adhesion between PMN and HUVEC increased dramatically(P