Evaluation of elastography combined with serological indexes for hepatic fibrosis in patients with chronic hepatitis B

AIM To investigate the value of ultrasound elastography combined with serological indexes in diagnosing liver fibrosis and assessing its severity.METHODS A total of 338 chronic hepatitis B(CHB) patients were divided into a disease group(patients with hepatic fibrosis) and control group(subjects without hepatic fibrosis). The disease group was further divided into S1-S4 according to the degree of fibrosis. Independent risk factors for hepatic fibrosis were analyzed using multivariate logistic regression. The diagnostic values of hepatic fibrosis from different indicators were compared using receiver operating characteristic(ROC) curves. The combination of elastography and serological indexes was explored to assess the severity of hepatic fibrosis.RESULTS The multivariate logistic regression analysis results revealed that shear wave velocity(SWV), hyaluronic acid(HA), type Ⅳ collagen(CⅣ) and aspartate aminotransferase-to-platelet ratio index(APRI) significantly affected the occurrence of hepatic fibrosis. The ROC curve revealed that the accuracy of the diagnosis of hepatic fibrosis for SWV and HA were 87.3% and 84.8%, respectively. The accuracy of SWV combined with HA was 88.9%. The multiple linear regression analysis revealed that SWV, aspartate aminotransferase(AST)/alanine aminotransferase(ALT), HA, CⅣ, APRI and fibrosis index based on the 4 factor(FIB-4) were screened as statistically significant independent factors. The established regression equation was: Fibrosis level =-4.046 + 1.024 × SWV + 1.170 × AST/ALT + 0.011 × HA + 0.020 × CⅣ + 0.719 × APRI + 0.379 × FIB-4. CONCLUSION SWV combined with serological indexes can improve the accuracy of diagnosis for CHB hepatic fibrosis. Serum indexes can help diagnose the degree of hepatic fibrosis.     

CD4+CD25+Foxp3+调节性T细胞在不同月龄小鼠中的变化及与肺癌关系

目的 探讨CD4+CD25+Foxp3+调节性T(Treg)细胞在衰老过程中的变化及与肺癌的关系.方法 建立Lewis肺癌模型,36只C57BL/6小鼠分成6组,青年健康组、中年健康组、老年健康组以及青年肿瘤组、中年肿瘤组、老年肿瘤组.通过流式细胞分析法测定6组小鼠脾脏细胞中CD4+CD25+Foxp3+Treg占CD4+T细胞的百分比来反映CD4+CD25+Foxp3+Treg细胞含量,通过实时荧光定量PCR法测定Foxp3mRNA的含量.结果 与健康组相比,肺肿瘤鼠脾脏中CD4+CD25+Foxp3+/CD4+T细胞和Foxp3 mRNA的含量明显增高(均P＜0.05);在健康组内,各年龄段CD4+CD25+Foxp3+/CD4+T细胞(F=47.70,P=0.000)和Foxp3mRNA(F=6.56,P=0.009)差异有统计学意义,老年鼠脾脏细胞中含有高数量的CD4+CD25+Foxp3+Treg细胞和Foxp3mRNA,最高组是老年肺肿瘤鼠.结论 CD4+CD25+Foxp3+Treg细胞和其功能基因Foxp3含量的改变与增龄和肺肿瘤的发生和发展存在密切的关系.

Abstract：

Objective To explore the change of CD4+CD25+Foxp3+ regulatory T (Treg) cells during aging and the relation with lung tumor. Methods The Lewis lung cancer model was set up in C57BL/6 female mice, and the 36 mice were divided into young health group, middle-aged health group, elderly health group, young tumor group, middle-aged tumor group and elderly tumor group. The percentages of CD4+CD25+Foxp3+ Treg in CD4+ T cells in mice spleen cells were measured by flow cytometry, for reflecting the quantity of CD4+CD25+Foxp3+ Treg cells. And the level of Foxp3 mRNA in splenocyte was tested by real-time PCR method. Results The level of CD4+CD25+Foxp3+/CD4+ T cells and the quantity of Foxp3 mRNA were higher in tumor groups than in healthy groups(both P＜0.05 ). Besides, in the healthy groups, there were statistical differences in the level of CD4+CD25+Foxp3+/CD4+ T cells (F=47.70, P=0.000) and the quantity of Foxp3 mRNA among the different months groups. Accumulation of the CD4+CD25+Foxp3+ Treg cells was accompanied with aging, the elderly mice contained a significantly larger population of CD4+CD25+Foxp3+ Treg cells in their spleen when compared with the younger counterparts, and the highest was the elderly tumor group. So it was with the functional gene Foxp3 mRNA (F=6.56, P=0.009). Conclusions The results suggest a close relationship of the change of CD4+CD25+Foxp3+Treg cells with aging and the genesis and development of lung tumor.     

Therapeutic effects and molecular mechanisms of anti-fibrosis herbs and selenium on rats with hepatic fibrosis

AIM:To study the therapeutic effects of anti-fibrosis herbsand selenium on hepatic fibrosis induced by carbontetrachloride (CCl_4) in rats and the underlining molecularmechanisms.METHODS:Fifty-three Wistar rats were randomly dividedinto:normal control group,model control group,colchicinegroup,anti-fibrosis herbs group (AF group) and anti-fibrosisherbs plus selenium group (AS group).The last four groupswere administered with CCl_4 at the beginning of experimentto induce hepatic fibrosis.Then colchicine,anti-fibrosis herbsand selenium were used to treat them.The normal controlgroup and the model control group were given normal salineat the same time.At the end of the 6~(th) week,rats in eachgroup were sacrificed.Blood and tissue specimens weretaken.Serum indicators (ALT,AST,HA,LN) were determinedand histopathological changes were graded.Lymphocyte CD_4and CD_8 were examined by flow cytometry.Expression ofTGF-β_1 and NF-κB was detected by immunohistochemistryand expression of TGF-β_1 mRNA was detected by semi-quantified RT-PCR.RESULTS:Histological grading showed much a smallerdegree of hepatic fibrogenesis in AS group and AF groupthan that in colchicine group and model control group.Theserum content of ALT,AST,HA and LN in AF group and ASgroup were significantly lower than that in colchicine group(ALT:65.8±26.5,67.3±18.4 and 96.2±20.9 in AF,AS andcolchicine groups respectively;AST:150.8±34.0,154.6±27.3and 215.8±24.6 respectively;HA:228±83,216±58 and416±135 respectively;LN:85.9±15.0,80.6±18.6 and106.3±14.2 respectively) (P<0.05).The level of CD_4 andCD_4/CD_8 ratio in AF group and AS group was significantly higherthat those in cochicine group (CD_4:50.8±3.8,52.6±3.4 and40.2±2.1 in AF,AS and colchicine groups respectively;CD_4/CD_8 ratio:1.45,1.46 and 1.26,respectively (P<0.05).The expression level of NF-κB and TGF-β_1 in the liver tissuesof AF and AS treatment groups was markedly decreasedcompared with that in cochicine group,and TGF-β_1 mRNAwas also markedly decreased (1.07±0.31 and 0.98±0.14 vs2.34±0.43,P<0.05).CONCLUSION:Anti-fibrosis herbs and selenium havebeneficial effects on hepatic fibrosis in rats by enhancingimmunity and inhibiting NF-κB and TGF-β_1 expressions.     

T cell immune response is correlated with fibrosis and inflammatory activity in hepatitis B cirrhotics

AIM: To explore the relationship among interferon-γ(IFN-γ) activity, fibrogenesis, T cell immune responses and hepatic inflammatory activity. METHODS: Peripheral blood samples from a total of 43 hepatitis B cirrhotic patients (LC) and 19 healthy controls (NC) were collected to measure their serum levels of IFN-γ, interleukin-2 (IL-2), soluble interleukin-2 receptor (sIL-2R), interleukin-10 (IL-10) and three serological markers of fibrosis including hyaluronic acid (HA), procol-lagen typeⅢpeptide (PⅢP), and typeⅣcollagen were measured using a double antibody sandwich ELISA. Also, serum total bilirubin (TB) and alanine aminotransferase (ALT) were measured by routine measures. RESULTS: The concentrations of serological markers of fibrosis in patients with active cirrhosis (ALC) were significantly higher than those in stationary liver cirrhosis (SLC) or NC groups. The levels of serological markers in HBeAg-positive patients were significantly higher than those in HBeAg-negative patients. In SLC and ALC patients, a negative linear correlation was found between IFN-γlevels and the serological markers of fibrosis. IFN-γand IL-2 levels in the ALC group were significantly higher than those in the SLC and NC groups, but the statistical difference was not significant between the latter two. In contrast, IL-10 levels in the SLC group were significantly higher than that in the NC group, but no significant difference was found between SLC and ALC groups. The sIL-2R level was elevated gradually in all these groups, and the differences were significant. Positive linear correlations were seen between IFN-γactivity and ALT levels (r=0.339, P < 0.05), and IL-2 activity and TB levels (r=0.517, P < 0.05). SIL-2R expression was positively correlated with both ALT and TB levels (r=0.324, 0.455, P < 0.05), whereas there was no statistically significant correlation between IL-10 expression and serum ALT and TB levels (r = -0.102, -0.093, P > 0.05). Finally, there was a positive correlation between IFN-γand IL-2 levels. CONCLUSION: T cell immune responses are correlated with fibrosis and hepatic inflammatory activity and may play an important role in liver cirrhosis.     

Multicenter clinical study on Fuzhenghuayu capsule against liver fibrosis due to chronic hepatitis B

AIM: To study the efficacy and safety of Fuzhenghuayu capsule (FZHY capsule, a capsule for strengthening body resistance to remove blood stasis) against liver fibrosis due to chronic hepatitis B. METHODS: Multicenter, randomized, double blinded and parallel control experiment was conducted in patients (aged from 18 to 65 years) with liver fibrosis due to chronic hepatitis B. Hepatic histologic changes and HBV markers were examined at wk 0 and 24 during treatment. Serologic parameters (HA, LM, P-Ⅲ-P, Ⅳ-C) were determined and B ultrasound examination of the spleen and liver was performed at wk 0,12 and 24. Liver function (liver function and serologic parameters for liver fibrosis) was observed at wk 0, 6, 12, 18 and 24. Blood and urine routine test, renal function and ECG were examined before and after treatment. RESULTS: There was no significant difference between experimental group (110 cases) and control group (106 cases) in demographic features, vital signs, course of illness, history for drug anaphylaxis and previous therapy, liver function, serologic parameters for liver fibrosis, liver histologic examination (99 cases in experimental group, 96 cases in control group), HBV markers, and renal function. According to the criteria for liver fibrosis staging, mean score of fibrotic stage(s) in experimental group after treatment (1.80) decreased significantly compared to the previous treatment (2.33, P<0.05), but there was no significant difference in mean score of fibrotic stage(s) (2.11 and 2.14 respectively). There was a significant difference in reverse rate between experimental group (52%) and control group (23.3%) in liver biopsy. With marked effect on decreasing the mean value of inflammatory activity and score of inflammation (P<0.05), Fuzhenghuayu capsule had rather good effects on inhibiting inflammatory activity and was superior to that of Heluoshugan capsule. Compared to that of pretreatment, there was a significant decrease in HA, LM, P-Ⅲ-P and Ⅳ-C content in experimental group after 12 and 24 wk of treatment. The difference in HA, LM, P-Ⅲ-P and Ⅳ-C content between 12 and 24 wk of treatment and pretreatment in experimental group was significantly greater than that in control group (P<0.01-0.05). The effect, defined as two of four parameters lowering more than 30% of the baseline, was 72.7% in experimental group and 27.4% in control group (P<0.01). Obvious improvement in serum Alb, ALT, AST and GGT was seen in two groups. Compared to that of control group, marked improvement in GGT and Alb was seen in experimental group (P<0.05). The effective rate of improvement in serum ALT was 72.7% in experimental group and 59.4% in control group. No significant difference was seen in blood and urine routine and ECG before and after treatment. There was also no significant difference in stable rate in ALT and serologic parameters for liver fibrosis between experimental group and control group after 12 wk of withdrawal. CONCLUSION: Fuzhenghuayu capsule has good therapeutic effects on alleviating liver fibrosis due to chronic hepatitis B without any adverse effect and is superior to that of Heluoshugan capsule.     

Effects of MSCs on the progression of atherosclerosis plaque in ApoE-knockout miceA

Background Immune inflammatory response is throughout the entire process of atherosclerosis(AS).It was unclear whether the mechanism of mesenchymal stem cells(MSCs)transplantation for treatment of AS is involved with inflammation regulation in the plaque area.The aim of this study was to explore the effects of MSCs in the formation of atherosclerosis plaque in hypercholesterolemic apoliprotein(apo)E/ mice.Methods ApoE/ mice MSCs were isolated and identified.At 8 weeks of age,30 male ApoE/ mice were randomly divided into negative control group(Neg,n = 10),positive control group(Pos,n = 10)and mesenchymal stem cells group(MSCs,n = 10).MSCs were injected through caudal vein into the body of Pos and MSCs group.The plaque area of all subjects were compared,the percentage of CD4 + CD25 + Tregs in different tissues were analyzed by fluorescence activated cell sorter(FACS),proliferation response of splenocytes to MSCs was detected and cytokines in the supernatant were determined by enzyme linked immunosorbent assay(ELISA).Results Compared with controls,MSCs resulted in a significant decrease of 1atherosclerotic plaques size(P < 0.05),and a significant increase of CD4 + CD25 + regulatory T cells in spleen(P < 0.05).Specific proliferation response of CD4 + CD25 + regulatory T cells in splenocytes to MCSs was significantly suppressed,the superanant level of TGF-β and IL-10 in MSCs group were increased while IFN-γ decreased significantly.Conclusion MSCs play an important role in regulating the inflammatory response and significantly inhibit the formation of the atherosclerosis plaque in ApoE/ mice.     

Effect of Chinese medicine Qinggan Huoxuefang on inducing HSC apoptosis in alcoholic liver fibrosis rats

AIM: To investigate the effect of Qinggan Huoxuefang (QGHXF) on improvement of liver function and pathology in rats, and to analyze the mechanism. METHODS: Wistar rats were divided into three groups at random: normal control group (12), micro-amount carbon tetrachloride group (CCl4)(12) and model group A (60). The model group A was ingested with the mixture (500 mL/L alcohol, 8 mL/kg per day; corn oil, 2 mL/kg per day; pyrazole, 24 mg/kg per day) once a day and intraperitoneal injections of 0.25 mL/kg of a 250 mL/L solution of CCl4 in olive oil twice a week for 12 wk. The CCl4 group received intraperitoneal injections only. At the end of 8 wk the model group A (60) was divided into 5 subgroups: model group, Xiaochaihu Chongji (XCH) group, QGHXF high dose group, moderate dose group and low dose group, and were given the drugs respectively. At the end of 12 wk, all the rats were killed and blood samples collected, as well as liver tissue. Blood samples were used for evaluation of alanine transaminase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma-glutamyltransferase (γ-GT). Liver specimens were obtained for routine HE, apoptosis gene array and flow cytometry analysis. RESULTS: A liver fibrosis animal model was successfully established. Fibrosis was obviously reduced in QGHXF high dose group, and no fibrosis formed in CCl4 group. Compared with model group the QGHXF group and XCH group could obviously decrease the level of ALT, AST, ALP, and GGT (P<0.05). QGHXF high dose group was better than XCH group in ALT (615±190 vs 867±115), and AST(1972±366 vs 2777±608). Moreover, QGHXF could reduce liver inflammation, fibrosis-induced hepatic stellate cell (HSC) apoptosis and regulate apoptosis gene expression. The HSC apoptosis rates of QGHXF groups were 22.4±3.13, 13.79±2.26 and 10.07±1.14, higher than model group, 6.58±1.04 (P< 0.05). Compared to model group, 39 genes were up-regulated, 11 solely expressed and 17 down-regulated in high dose group. CONCLUSION: QGHXF can improve liver fibrosis and induce HSC apoptosis.     

4-1BB单克隆抗体对免疫性肝损伤治疗及CD4+CD25+T淋巴细胞影响研究

目的 探讨4-1BB单克隆抗体(4-1BBmAb)对免疫性肝损伤治疗及对CD4+CD25+T淋巴细胞影响.方法 建立小鼠刀豆蛋白A(ConA)肝损伤模型,检测4-1BB表达,ConA注射后2 h给予4-1BBmAb(100μg/只),观察肝功能及病理学变化,流式细胞仪检测CD4+CD25+T淋巴细胞.结果 丙氨酸转氨酶(ALT)模型组(139±22)U/L,对照组(32±12)U/L,天冬氨酸转氨酶(AST)分别(130±16)U/L及(29±11)U/L,两组差异有统计学意义(P<0.01);模型组4-1BB表达(8.1±2.6)比对照组(5.3±2.6)升高(P<0.01).4-1BBmAb治疗后ALT(98±14)U/L,AST(89±11)U/L降低(P<0.01).模型组CD4+CD25+T淋巴细胞(2.9±0.8)低于对照组(3.6±1.2)(P<0.05),4-1BBmAb组(8.3±3.0)高于生理盐水组(3.0±0.8)(P<0.01).结论 4-1BBmAb对免疫性肝损伤有治疗作用,影响CD4+CD25+T淋巴细胞达到治疗肝损伤目的 .     

阿苯达唑联合黄芪对华支睾吸虫感染大鼠肝功能异常的疗效观察

目的 观察阿苯达唑联合黄芪对华支睾吸虫感染大鼠肝功能异常的疗效.方法 健康成年Wistar大鼠32只,按体质量随机分为对照组、未治疗组、阿苯达唑组和阿苯达唑+黄芪组.每组8只.未治疗组、阿苯达唑组和阿苯达唑+黄芪组大鼠给华支睾吸虫囊蚴50个/只灌胃,对照组大鼠给生理盐水灌胃:阿苯达唑组大鼠在感染后10周每日每只给阿苯达唑50 mg/kg灌胃,连续5 d;阿苯达唑+黄芪组大鼠在给阿苯达唑的同时腹腔注射黄芪注射液800 mg/kg,连续30 d.于感染后14周采集血样,检测血清丙氨酸转氨酶(ALT)、总胆红素(TBIL)、透明质酸酶(HA)、层粘连蛋白(LN)和Ⅲ型前胶原(PCⅢP)水平.结果 大鼠血清ALT和LN水平组间比较,差异有统计学意义(F值分别为31.40、11.82,P<0.01).血清ALT和LN水平,未治疗组[(85.50±9.65)U/L、(64.20±4.18)μg/L]、阿苯达唑组[(65.29±7.78)U/L、(58.23±2.55)μg/L]较对照组[(47.88±4.88)U/L、(51.20±4.12)μg/L]明显升高(P<0.05);阿苯达唑组和阿苯达唑+黄芪组[(50.25±9.29)U/L、(53.68±5.63)μg/L]较未治疗组明显下降(P<0.05),阿苯达唑+黄芪组下降较阿苯达唑组更明显(P<0.05).大鼠血清TBIL、PⅢNP和HA组间比较差异有统计学意义(χ2值分别是15.309、21.418、19.759,P<0.01).血清TBIL、PⅢNP、HA水平,未治疗组(2.400 μmol/L、46.220 μg/L、310.885 μg/L)、阿苯达唑组(1.200 μmol/L、36.540 μg/,L、178.010 μg/L)较对照组(0.700 μmol/L、26.085 μg/L、81.935 μg/L)明显升高(P<0.05).大鼠血清TBIL水平,阿苯达唑+黄芪组(0.750 μmol/L)较未治疗组明显降低(P<0.05):大鼠血清PⅢNP和HA水平,阿苯达唑组和阿苯达唑+黄芪组(30.470 μg/L、100.240 μg/L)较未治疗组明显降低(P<0.05),与阿苯达唑组比较,阿苯达唑+黄芪组大鼠血清TBIL、pⅢNP和HA水平下降更明显(P<0.05).结论 大鼠感染华支睾吸虫后肝功能异常,经阿苯达唑联合黄芪治疗后肝功能得到改善,肝纤维化减轻,阿苯达唑联合黄芪治疗效果好于单独应用阿苯达唑治疗.     

CD4+ CD25+ CD127dim/-调节性T淋巴细胞对肝星状细胞增殖及功能的影响

目的 了解CD4+ CD25+ CD127dim/-调节性T淋巴细胞在体外对肝星状细胞(HSC)增殖以及功能的影响,初步探讨调节性T淋巴细胞促肝纤维化的机制。方法传代培养HSC LX-2,将免疫磁珠细胞分选(MASC)法分离所得慢性乙型肝炎患者调节性T淋巴细胞与HSC LX-2按不同方式共培养5d,以单独培养的HSC作为对照,细胞计数试剂盒-8（CCK-8）法检测共培养HSC增殖情况,ELISA法检测上清液中转化生长因子(TGF)β1含量,放射免疫法检测HSC分泌HA、PCⅢ水平。统计学处理采用LSD-t检验。结果 5例调节性T淋巴细胞与HSC比例为1.5∶1时HSC增殖最为明显,10例直接接触共培养与使用Transwell系统共培养调节性T淋巴细胞与HSC吸光度值分别为(0.713±0.032)、(0.735±0.028) cpm,均较对照组的（0.677±0.029）cpm增殖明显(t=5.4003,8.7878;均P＜0.01)。10例直接接触共培养与Transwell组细胞上清液中TGFβ1浓度分别为(781.59±76.45)、（813.53±60.62）pg/mL,显著高于对照组的(722.51±59.66) pg/mL（t=4.0014,6.1653;均P＜0.01）;HA浓度分别为（433.57±27.90）、（445.40±23.73）ng/mL,显著高于对照组的（415.83±19.44）ng/mL（t=3.3124,5.5231;均P＜0.01）;PCⅢ浓度分别为(21.93±1.71)、(23.12±1.87) ng/mL,显著高于对照组的（20.10±1.49）ng/mL(f=4.8082,7.9436;均P＜0.01)。且Transwell组各项结果均显著高于直接接触组(t=3.3875,2.1639,2.2107,3.1354;均P＜0.05)。结论CD4+ CD25+ CD127dim调节性T淋巴细胞可促进共培养的HSC增殖及其HA、PCⅢ的分泌。体外实验证明,CD4+ CD25+ CD127dim/-调节性T淋巴细胞具有促进肝纤维化的重要作用。     

晚期血吸虫病肝纤维化患者血清维生素D水平与免疫失衡的关系

目的 探索晚期血吸虫病肝纤维化患者血清维生素D水平与免疫失衡间的关系.方法 选择2016年5月至2018年9月就诊于嘉兴市第一医院血吸虫病科的120例晚期血吸虫病肝纤维化患者作为观察组,选取50例同期该院健康体检者作为对照组,比较两组血清中IgG抗体、IgA抗体、C3补体、C4补体、CD4+细胞比例、CD8+细胞比例、...     

愈肝龙胶囊联合恩替卡韦治疗乙型肝炎肝纤维化临床观察

目的:研究愈肝龙胶囊联合恩替卡韦治疗慢性乙型肝炎肝纤维化的临床疗效。方法:将符合纳入标准的116例慢性乙型肝炎肝纤维化患者随机分为观察组64例和对照组52例,两组患者均给予恩替卡韦抗病毒治疗,观察组加用愈肝龙胶囊。观察两组患者治疗前与治疗36个月后肝硬度值(LSM)、超声影像学指标(门静脉内径、脾脏厚度)、血清肝纤维化指标4项(HA、LN、C-Ⅳ、PC-Ⅲ)、肝功能(ALT、AST、PLT、Alb、TBil)、APRI指数的变化。结果:两组患者治疗36个月后,观察组的LSM值、肝纤维化指标(C-Ⅳ)、肝功能(ALT、AST、PLT、Alb、TBil)以及APRI指数优于治疗前(P<0.05),且与对照组比较差异有显著性意义(P<0.05)。结论:复方中成药愈肝龙胶囊联合恩替卡韦改善慢性乙型肝炎肝患者肝纤维化与肝功能的效果优于单用恩替卡韦。     

慢性乙肝患者HBV-DNA、肝功能指标与肝纤维化的关系

[目的]探究慢性乙型病毒性肝炎患者乙肝病毒脱氧核糖核酸(HBV-DNA)、肝功能指标与肝纤维化的关系.[方法]入选65例慢性乙肝患者为患者组;同期选取肝功能合格的体检者65例作为对照组;检测2组肝纤维化指标[血清透明质酸(HA)、层粘连蛋白(LN)、Ⅲ型胶原(PCⅢ)、Ⅳ型胶原(Ⅳ-C)],HBV-DNA,肝功能指标[...     

支气管哮喘大鼠CD4+CD25+T细胞的变化及地塞米松干预作用的研究

目的 研究支气管哮喘(简称哮喘)大鼠模型支气管肺泡灌洗液(BALF)、血液、脾脏CD4+CD25+T细胞的变化,及地塞米松对CD4+CD25+T细胞的影响.方法 50只SD大鼠随机分为5组,空白对照(A)组,哮喘(B)组,地塞米松1(C)组、地塞米松2(D)组,地塞米松3(E)组.A组第l天给予腹腔注射生理盐水l ml,第15～21天每天给予生理盐水雾化.B、C、D、E组用卵蛋白建立哮喘大鼠模型,第1天,每只大鼠腹腔注射抗原l ml(卵蛋白1 mg+灭活百日咳杆菌9×106个+氢氧化铝干粉100 mg)混悬液,第15～21天给予1%的卵蛋白雾化30 min,C、D、E组于雾化后分别给予腹腔注射地塞米松0.2 mg/kg、1 mg/kg、2 mg/kg.采用流式细胞仪检测的方法 ,观察大鼠体内BALF、外周血、脾脏CD4+CD25+T细胞的变化及使用不同剂量地塞米松后对其的影响.结果 B组BALF、外周血、脾脏CD4+CD25+T细胞表达占CD4+T细胞的百分比分别是(42.21±5.62)%、(12.69±2.70)%、(11.15±1.05)%,A组结果 分别是(18.76±5.85)%、(6.21±1.73)%、(7.85±2.13)%.B组与A组比较,差异均具有统计学意义(P＜0.01,P＜0.01,P＜0.05);C组、D组、E组BALF中CD4+CD25+T细胞占CD4+T细胞的百分比表达分别是(10.49±4.03)%、(13.28±5.12)%、(7.51±5.39)%,显著低于A组和B组,(P＜0.05,P＜0.01);外周血中,C组(6.03±1.43)%、D组(4.88±0.95)%与A组(6.21±1.73)%比较,差异无统计学意义,E组(3.49±0.62)%与C组、A组比较,差异有统计学意义(P＜0.05).脾脏中,C组(7.25±1.82)%、D组(8.63±3.18)%与A组(7.85±2.13)%比较,差异无统计学意义,E组(3.38±1.37)%与C组、D组、A组比较,差异有统计学意义(P＜0.05).结论 CD4+CD25+T细胞在哮喘大鼠体内有明显的优势表达,可能是哮喘发病的机制之一.地塞米松可以抑制CD4+CD25+T细胞的表达.BALF内CD4+CD25+T细胞的变化与外周血和脾脏的变化具有一致性,监测外周血或脾脏CD4+CD25+T细胞变化可了解肺部情况.     

扩张型心肌病患者CD4~+CD25~+Foxp3~+ T细胞的检测及意义

目的:探讨扩张型心肌病(DCM)患者外周血CD4+CD25+Foxp3+T细胞的水平及意义。方法:采用流式细胞术检测DCM患者30例及健康对照组20例外周血CD4+CD25+T细胞和CD4+CD25+Foxp3+T细胞的比例。结果:DCM患者外周血CD4+CD25+T细胞占CD4+T细胞的比例为(8.53±1.64)%,显著低于健康对照组的(11.4±2.17)%,P0.01;DCM患者CD4+CD25+Foxp3+T细胞占CD4+T细胞比例为(0.99±0.54)%,显著低于健康对照组的(1.55±0.55)%,P0.01;且DCM患者心功能越差,CD4+CD25+Foxp3+T细胞占CD4+T细胞的比例越低。结论:DCM患者调节性T细胞比例的减少,可能打破了自身免疫耐受,发生了针对心肌抗原的自身免疫反应,参与了DCM的发病。     

CD34+细胞水平变化对老年人不同程度慢性左心衰竭的影响

目的 观察循环CD34+细胞(CD34+)水平变化对不同程度老年慢性左心衰竭的影响.方法 根据美国纽约心脏病学会(NYHA)分级临床标准和左室射血分数(LVEF)将所有入选者分为Ⅰ级组23例,Ⅱ级组27例,Ⅲ级组20例,Ⅳ级组16例,同期健康对照组41例.测定不同程度老年慢性左心衰竭患者CD34+的水平,并与肿瘤坏死因子-α(TNF-α)、可溶性肿瘤坏死因子受体1、2(sTNFR-1 sTNFR一2)和血管内皮生长因子(VEGF)水平进行对照分析.结果 慢性左心衰竭的早期CD34+水平升高,随着慢性左心衰竭程度的加重CD34+的水平下降,对照组、NYHA I组、NYHAⅡ组、NYHAⅢ组和NYHAⅣ组分别为(0.6±0.2)×108/L、(2.4±0.4)×109/L、(1.9±0.2)×109/L、(1.3±0.1)×109/L和(0.5±0.2)×109/L,两两比较差异有统计学意义(均P<0.01).TNF-α,、sTNFR-1、sTNFR-2和VEGF水平明显增高,NYHA Ⅰ组与NYHAⅣ组TNF-α[分别为(28.4±10.8)ng/L与(61.4±15.7)ng/L]、sTNFR-1[(690.8±62.7)ng/L与(2820.9±1282.8)ng/L]、sTNFR_2[(740.8±112.3)ng/L与(4113.1±1102.2)ng/L]、VEGF [(423.3±147.9)ng/L与(996.3±487.1)ng/L]比较,差异有统计学意义(均P<0.01).结论 CD34+水平的变化可能预测老年慢性左心衰竭的发生、发展及严重程度.     

外源性尿激酶抗肝纤维化的作用机制

目的 探讨外源性尿激酶抗大鼠肝纤维化的作用机制.方法 采用复合致病因子复制肝纤维化大鼠模型.大鼠随机分为对照组(正常饮食)、肝纤维化组(复合致病因子饲养6周)和尿激酶预防组(复合致病因子+尿激酶饲养6周).检测并比较各组大鼠血透明质酸含量、肝组织内羟脯氨酸、α-平滑肌肌动蛋白(α-SMA)、金属蛋白酶组织抑制因子-1(TIMP-1)、尿激酶型纤溶酶原激活物(μPA).尿激酶型纤溶酶原激活物抑制剂-1(PAI-1)、转化生长因子β1(TGF β 1)、Ⅰ型胶原蛋白和Ⅲ型胶原蛋白表达量以及PAI-1 mRNA和TGF β 1 mRNA的相对表达量.多组间用单因素方差分析q检验,两组间比较采用t检验,多组间等级资料比较采用Kruskal Wallis H检验,多样本间两两比较采用扩展的t检验. 结果血浆ALT、AST、总胆红素、透明质酸和肝组织内羟脯氨酸含量在尿激酶预防组大鼠分别为(46.66±6.30)U/L、(126.26±31.65)U/L、(31.11±4.20)μmol/L、(109.70±18.81)μg/L和(0.98±0.09)mg/g,较肝纤维化组的(101.57±11.97)U/L、(205.89±56.26)U/L、(67.75±2.75)μmol/L、(184.43±32.36)μg/L和(1.65±0.16)mg/g均明显降低(q值分别为3.3801～20.0061,P值均<0.01).尿激酶预防组α-SMA、Ⅰ型胶原蛋白、Ⅲ型胶原蛋白、TIMP-1、PAI-1及TGF β 1蛋白相对表达量分别为299.27±37.36、210.05±27.17、192.94±24.48、213.70±32.21、204.25±17.92和205.97±23.81,较肝纤维化组的418.83±30.21、323.77±21.53、302.37±31.43、376.63±25.19、313.53±26.67和327.42±36.75均明显减少,而uPA蛋白表达增加.尿激酶预防组PAI-1 mRNA,TGF β 1 mRNA表达减少,肝纤维化程度明显减轻. 结论 预防性使用外源性尿激酶可减轻肝损伤,降低血浆转氨酶、胆红素,减少肝星状细胞活化,降低TIMP-1、PAI-1蛋白表达,增加uPA蛋白表达,加速组织损伤修复,延缓肝纤维化的发生.