High densities of serotonin and peptide YY cells in the colon of patients with lymphocytic colitis

AIM:To investigate colonic endocrine cells in lymphocytic colitis(LC) patients.METHODS:Fifty-seven patients with LC were included.These patients were 41 females and 16 males,with an average age of 49 years(range 19-84 years).Twenty-seven subjects that underwent colonoscopy with biopsies were used as controls.These subjects underwent colonoscopy because of gastrointestinal bleeding or health worries,where the source of bleeding was identified as haemorrhoids or angiodysplasia.They were 19 females and 8 males with an average age of 49 years(range 18-67 years).Biopsies from the right and left colon were obtained from both patients and controls during colonoscopy.Biopsies were fixed in 4% buffered paraformaldehyde,embedded in paraffin and cut into 5 m-thick sections.The sections immunostained by the avidin-biotin-complex method for serotonin,peptide YY(PYY),pancreatic polypeptide(PP) enteroglucagon and somatostatin cells.The cell densities were quantified by computerised image analysis using Olympus software.RESULTS:The colon of both the patient and the control subjects were macroscopically normal.Histopathological examination of colon biopsies from controls revealed normal histology.All patients fulfilled the diagnosis criteria required for of LC:an increase in intraepithelial lymphocytes( 20 lymphocytes/100 epithelial cells) and surface epithelial damage with increased lamina propria plasma cells and absent or minimal crypt architectural distribution.In the colon of both patients and control subjects,serotonin-,PYY-,PP-,enteroglucagon-and somatostatin-immunoreactive cells were primarily located in the upper part of the crypts of Lieberkühn.These cells were basket-or flask-shaped.There was no statistically significant difference between the right and left colon in controls with regards to the densities of serotonin-and PYYimmunoreactive cells(P = 0.9 and 0.1,respectively).Serotonin cell density in the right colon in controls was 28.9 ± 1.8 and in LC patients 41.6 ± 2.6(P = 0.008).In the left colon,the corresponding figures were 28.5 ± 1.9 and 42.4 ± 2.9,respectively(P = 0.009).PYY cell density in the right colon of the controls was 10.1 ± 1 and of LC patients 41 ± 4(P = 0.00006).In the left colon,PYY cell density in controls was 6.6 ± 1.2 and in LC patients 53.3 ± 4.6(P = 0.00007).CONCLUSION:The change in serotonin cells could be caused by an interaction between immune cells and serotonin cells,and that of PYY density might be secondary.     

慢性乙型肝炎病毒感染者外周血单个核细胞表达程序性死亡分子-1及其配体与血清乙型肝炎病毒DNA水平的相关性

Objective To study the relationship between programmed death-1 (PD-1)/programmed death-1 ligand (PD-L1) expressions and serum hepatitis B virus (HBV) DNA levels in chronic hepatitis B (CHB) patients. Methods A total of 137 CHB patients and 10 healthy controls were enrolled in the study. The peripheral blood mononuclear cells (PBMCs) were isolated from fresh blood samples. HBV-specific cytotoxic T lymphocyte (CTL) was expanded in vitro in 64 human leucocyte antigen (HLA)-A2 positive patients. Flow cytometry was used to detect HLA-A2 type,expressions of PD-1/PD-L1 on PBMCs and PD-1 on HBV specific CTL. Interferon gamma (IFN-γ)was measured by commercial enzyme-linked immunosorbent assay (ELISA) kits. PD-1/PD-L1expressions on PBMCs, HBV-specific CTL and IFN-γ level in PBMC culture medium were compared among patients with different baseline HBV DNA levels. Ten hepatitis B e antigen (HBeAg) positive patients were treated with telbivudine for 24 weeks. The above mentioned parameters were determined and compared before and after the antiviral treatment. Independent-samples t test were used to compare means between two groups and one-way A NOVA were used to compare means among multigroups. We used the pearson corretation test to assess corretation significance. Results The PD-1 and PD-L1 expressions on PBMCs in patients with baseline HBV DNA＜3 lg copy/mL, 3-6 lg copy/mL and ＞6 lg copy/mL were all significant higher than those in healthy control group, but no statistical differences were found. PD-1 expressions on HBV-specific CTL in the three CHB patient groups were (69.3±11.2)%, (76.5±9. 1)% and (78.0±11.7)%, respectively. However, PD-1 expression on HBV-specific CTL was higher, while the frequency of HBV-specific CTL cells was lower in HBV DNA ＞6 lg copy/mL group compared to HBV DNA＜3 lg copy/mL group. The above parameters, including expressions of PD-1 and PD-L1, the frequency of HBV-specific CTL and its PD-1 expression were not significantly different between HBeAg-positive group and HBeAg-negative group. Compared with baseline, PD-1 and PD-L1 expression decreased obviously accompanying with increase of HBV-specific CTL cells frequency and IFN-γ level after 12 weeks and 24 weeks of telbivudine treatment. Conclusions PD-1 expression on HBV-specific CTL correlates with serum HBV DNA level, but not HBeAg status in CHB patients. Suppression of HBV replication can reduce PD-1/PD-L1 expressions and partially restore HBV specific CTL function.     

慢性乙型肝炎病毒感染者外周血单个核细胞表达程序性死亡分子-1及其配体与血清乙型肝炎病毒DNA水平的相关性

Objective To study the relationship between programmed death-1 (PD-1)/programmed death-1 ligand (PD-L1) expressions and serum hepatitis B virus (HBV) DNA levels in chronic hepatitis B (CHB) patients. Methods A total of 137 CHB patients and 10 healthy controls were enrolled in the study. The peripheral blood mononuclear cells (PBMCs) were isolated from fresh blood samples. HBV-specific cytotoxic T lymphocyte (CTL) was expanded in vitro in 64 human leucocyte antigen (HLA)-A2 positive patients. Flow cytometry was used to detect HLA-A2 type,expressions of PD-1/PD-L1 on PBMCs and PD-1 on HBV specific CTL. Interferon gamma (IFN-γ)was measured by commercial enzyme-linked immunosorbent assay (ELISA) kits. PD-1/PD-L1expressions on PBMCs, HBV-specific CTL and IFN-γ level in PBMC culture medium were compared among patients with different baseline HBV DNA levels. Ten hepatitis B e antigen (HBeAg) positive patients were treated with telbivudine for 24 weeks. The above mentioned parameters were determined and compared before and after the antiviral treatment. Independent-samples t test were used to compare means between two groups and one-way A NOVA were used to compare means among multigroups. We used the pearson corretation test to assess corretation significance. Results The PD-1 and PD-L1 expressions on PBMCs in patients with baseline HBV DNA＜3 lg copy/mL, 3-6 lg copy/mL and ＞6 lg copy/mL were all significant higher than those in healthy control group, but no statistical differences were found. PD-1 expressions on HBV-specific CTL in the three CHB patient groups were (69.3±11.2)%, (76.5±9. 1)% and (78.0±11.7)%, respectively. However, PD-1 expression on HBV-specific CTL was higher, while the frequency of HBV-specific CTL cells was lower in HBV DNA ＞6 lg copy/mL group compared to HBV DNA＜3 lg copy/mL group. The above parameters, including expressions of PD-1 and PD-L1, the frequency of HBV-specific CTL and its PD-1 expression were not significantly different between HBeAg-positive group and HBeAg-negative group. Compared with baseline, PD-1 and PD-L1 expression decreased obviously accompanying with increase of HBV-specific CTL cells frequency and IFN-γ level after 12 weeks and 24 weeks of telbivudine treatment. Conclusions PD-1 expression on HBV-specific CTL correlates with serum HBV DNA level, but not HBeAg status in CHB patients. Suppression of HBV replication can reduce PD-1/PD-L1 expressions and partially restore HBV specific CTL function.     

慢性乙型肝炎患者树突状细胞功能与单个核细胞乙型肝炎病毒共价闭合环状DNA的关系

Objective To investigate the relationship between the maturity and function of dendritic cells (DC) and hepatitis B virus covalently closed circular DNA (HBV cccDNA) load in the peripheral blood mononuclear cells (PBMC)/monocyte-derived DC in patients with chronic hepatitis B (CHB). Methods The peripheral blood samples were collected from 29 patients with CHB and 10healthy controls. PBMC were isolated freshly and induced with granulocyte/macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4). A large amount of DC were harvested after seven days of culture. The expressions of CD209, CD80, CD86, human leucocyte antigen (HLA)-DR and CD1a of DC were analyzed by flow cytometry. The HBV cccDNA load in PBMC and DC were measured by real-time polymerase chain reaction (PCR). The interleukin-12 (IL-12) level in the culture supernatant of DC was determined by enzyme linked immunosorbent assay (ELISA). The effects on T lymphocyte proliferation induced by DC were tested by mixed lymphocyte reaction (MLR). The data was compared by t test and analysis of variance. Results HBV cccDNA could be detected in PBMC from 16 patients, but not in DC from all 29 patients. HBV cccDNA load was all negatively correlated with the expressions of CD209 (r= -0. 793, P＜0.01), CD80 (r= -0. 581,P＜0.05), CD86 (r=-0. 698, P＜0.01), HLA-DR (r=-0. 817, P＜0.01), CD1a (r=-0. 734, P＜0.01), IL-12 level (r=-0. 632, P＜0.05) and allogenic T lymphocyte proliferation induced by DC (r=-0. 617, P＜0.05). The expressions of CD209, CD80, CD86, CD1a and HLA-DR on DC,IL-12 level in culture supernatant of DC and the allogenic T lymphocyte proliferation induced by DC in patients with positive PBMC HBV cccDNA were all significantly lower compared to those in healthy controls, and the changes of the parameters mentioned above were greater in PBMC HBV cccDNA positive patients than those in PBMC HBV cccDNA negative patients (P ＜ 0. 05 or P ＜ 0. 01).Conclusions The function and maturity of DC are impaired in CHB patients. HBV cccDNA can be detected in PBMC from CHB patients. Moreover, the higher PBMC HBV cccDNA is, the worse DC function and maturity are, which could be one of the important mechanisms of HBV persistent infection.     

慢性乙型肝炎患者树突状细胞功能与单个核细胞乙型肝炎病毒共价闭合环状DNA的关系

Objective To investigate the relationship between the maturity and function of dendritic cells (DC) and hepatitis B virus covalently closed circular DNA (HBV cccDNA) load in the peripheral blood mononuclear cells (PBMC)/monocyte-derived DC in patients with chronic hepatitis B (CHB). Methods The peripheral blood samples were collected from 29 patients with CHB and 10healthy controls. PBMC were isolated freshly and induced with granulocyte/macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4). A large amount of DC were harvested after seven days of culture. The expressions of CD209, CD80, CD86, human leucocyte antigen (HLA)-DR and CD1a of DC were analyzed by flow cytometry. The HBV cccDNA load in PBMC and DC were measured by real-time polymerase chain reaction (PCR). The interleukin-12 (IL-12) level in the culture supernatant of DC was determined by enzyme linked immunosorbent assay (ELISA). The effects on T lymphocyte proliferation induced by DC were tested by mixed lymphocyte reaction (MLR). The data was compared by t test and analysis of variance. Results HBV cccDNA could be detected in PBMC from 16 patients, but not in DC from all 29 patients. HBV cccDNA load was all negatively correlated with the expressions of CD209 (r= -0. 793, P＜0.01), CD80 (r= -0. 581,P＜0.05), CD86 (r=-0. 698, P＜0.01), HLA-DR (r=-0. 817, P＜0.01), CD1a (r=-0. 734, P＜0.01), IL-12 level (r=-0. 632, P＜0.05) and allogenic T lymphocyte proliferation induced by DC (r=-0. 617, P＜0.05). The expressions of CD209, CD80, CD86, CD1a and HLA-DR on DC,IL-12 level in culture supernatant of DC and the allogenic T lymphocyte proliferation induced by DC in patients with positive PBMC HBV cccDNA were all significantly lower compared to those in healthy controls, and the changes of the parameters mentioned above were greater in PBMC HBV cccDNA positive patients than those in PBMC HBV cccDNA negative patients (P ＜ 0. 05 or P ＜ 0. 01).Conclusions The function and maturity of DC are impaired in CHB patients. HBV cccDNA can be detected in PBMC from CHB patients. Moreover, the higher PBMC HBV cccDNA is, the worse DC function and maturity are, which could be one of the important mechanisms of HBV persistent infection.     

Effect of Hejie decoction on T cell immune state of chronic hepatitis B patients

AIM: To explore the effect of Hejie decoction (HJD) (mediation decoction) on T cellular immune state of chronic hepatitis B patients.METHODS: Sixty-five patients with chronic hepatitis B were randomly divided into 2 groups. Forty patients in the treatment group were treated by HJD, and 25 patients in the control group were treated by routine Western medicine. The TCRVβ7 gene expression, T lymphocyte subsets (CD3^+, CD4^+, CD8^+,CD4^+/CD8^+) levels were observed before and after treatment.RESULTS: The level of CD4^+ cells was lower whereas the level of CD8^+ cells was higher in patients than in the normal group. There was no significant difference between the levels of CD3^+ cells in patients and normal persons. After 6 months of treatment, ALT, AST, TB levels of the 2 groups were obviously decreased, and the level of CD4^+ cells was increased whereas the level of CD8^+ cells was decreased in the treatment group. However, the level of CD4^+ cells and CD8^+ cells had no significant difference in the control group. TCRVβ7 expressions were detected in 6 patients of the treatment group, whose HBV-DNA and HBeAg turned negative and ALT became normal. HBeAg in another 3 patients turned negative while HBV-DNA did not, and TCRVβ7 expressions were not detectable. TCRVβ7 expression could not be detected in the control group, HBV-DNA of the control group did not turn negative. HBeAg in 1 patient turned negative while HBV-DNA did not, and TCRVβ7 expressions were not detectable. The total effective rate was not significantly different between the 2 groups and the markedly effective rate was significantly different(P<0.01).CONCLUSION: H3D is effective for treating chronic hepatitis B, and its effect seems to relate with the improvement of the TCRVβ7 expression of chronic hepatitis B patients, thus activating T cells and eliminating HBV. T cellular immune function plays an important role in HBV infection and virus elimination.     

Upregulated voltage-gated potassium channel Kv1.3 on CD4+CD28null T lymphocytes from patients with acute coronary syndrome

Objective The purpose of our study is to observe the voltage-gated potassium channel Kv1.3 expressed on CD4+ CD28null T cells from the peripheral blood of acute coronary syndrome (ACS) patients by the patch clamp technique. Methods Kvl.3 potassium channels expression from 17 patients with ACS and 11 healthy age-match controls was detected in single cell(CD4+CD28null T cells and CD4+CD28+T cells) by fluorescence microscopy and patch clamp. Results The percentage of CD4+CD28null T cells was higher in the ACS patients [(6.97±2.05)%] than that in the controls [(1.38±0.84)%, P<0.05]. The concentration of hsCRP was directly correlated with the number of the CD4+CD28null T cells in the ACS patients (r=0.52, P<0.05). The conductance (6.89±1.17ns vs 3.36±0.66ns), dens (1.95±0.80 μm2 vs 1.13±0.57 μm2) and numbers (574.5±97.6 n/cell vs. 280.3±55.3 n/cell) of the Kvl.3 channels on the CD4+CD28null T cells were significantly higher than those on the CD4+CD28+T cells (all P<0.0l) in ACS patients, but were similar on CD4+CD28+T between ACS patients and controls. Conclusion The CD4+CD28null T cells and the numbers of Kv1.3 channels on the CD4+CD28null T cells from patients with ACS are significantly upregulated and might contribute to the pathogenesis of ACS.     

Effects of smoking cessation on expressions of nuclear factor-κB, matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 in airway epithelial cells of smoking rats

Objective To investigate the effects of smoking and smoking cessation on airway inflammation and remodeling in chronic obstructive pulmonary diseases through detecting mRNA and protein expressions of nuclear factor-κB (NF-κB), cell matrix metalloproteinase-9 (MMP-9) and cellular tissue inhibitor of metalloproteinase-1 (TIMP-1) in airway epithelial cells of smoking and smoking cessation rats. Methods Twenty-four male Wistar rats were randomly divided into control group, smoking group and smoking cessation group,eight in each group. Hybridization in situ and immunohistochemistry were used to detect mRNA and protein expressions of NF-κB, MMP-9 and TIMP-1 in airway epithelial cells of rats. Results ① Compared with control group (0.29 ± 0.06,0.29±0.06), mRNA and protein expressions of NF-κB in smoking group (0.45±0.04,0.41±0.03) and smoking cessation group (0.40±0.05,0.37±0.03) were higher (all P＜0.05). The mRNA and protein expressions of NF-κB in smoking cessation group were lower than those in smoking group (all P ＜0.05). ②Compared with control group (0.30±0.06,0.30±0.06) ,mRNA and protein expressions of MMP-9 in smoking group (0.52±0.03,0.51±0.07) and smoking cessation group (0.38±0.03,0.33±0.02) were higher (all P＜0.05). The mRNA and protein expressions of MMP-9 in smoking cessation group were lower than those in smoking group (all P＜0.05). ③Compared with control group (0.26±0.04, 0.26±0.04), mRNA and protein expressions of TIMP-1 in smoking group (0.49±0.05,0.37±0.03) and smoking cessation group (0.42±0.04,0.35±0.03) were higher (all P ＜0.05). The mRNA and protein expressions of TIMP-1 in smoking cessation group were lower than those in smoking group (all P ＜ 0.05). ④ Compared with control group (1.00±0.02,1.00±0.02), MMP-9/TIMP-1 mRNA and protein expressions were larger than one in smoking group (1.07±0.14, 1.37±0.19), and less than one in smoking cessation group (0.92±0.13,0.94±0.10) (all P ＜0.05). ⑤The mRNA and protein expressions of NF-κB and MMP-9in each group were positively correlation (r=0.87,0.66,all P ＜0.05). Conclusions In airway epithelial cells of smoking rats, mRNA and protein expressions of NF-κB, MMP-9 and TIMP-1 increase, and MMP-9/TIMP-1 is larger than one. After stoping smoking, mRNA and protein expressions of NF-κB,MMP-9 and TIMP-1 decrease, and MMP-9/TIMP-1 is less than one. This experiment explains that smoking can cause airway inflammation and remodeling, smoking cessation can reduce airway inflammation and remodeling.     

母牛分枝杆菌菌苗改善老年慢性阻塞性肺疾病稳定期患者免疫功能的研究

目的 探讨母牛分枝杆菌菌苗改善老年慢性阻塞性肺疾病(COPD)稳定期患者免疫功能及临床意义.方法 选择老年COPD稳定期患者100例,按随机数字表法分为母牛分枝杆菌菌苗治疗组和常规治疗组各50例,另设健康对照组50例.分别测定菌苗治疗组及常规治疗组治疗前、后和健康对照组外周血T淋巴细胞亚群(CD3+、CD4+、CD8+、CD4+/CD8+)、自然杀伤细胞(NK细胞)、免疫球蛋白(IgG、IgA、IgM)、细胞因子[白介素-6(IL-6)、白介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)]水平.菌苗治疗组及常规治疗组随访1年,同时比较两组患者人均年急性加重次数和住院次数.结果 菌苗治疗组及常规治疗治疗前与健康对照组比较,CD3+、CD4+、CD4+/CD8+、NK细胞水平降低差异有统计学意义(t值分别为8.09和8.09、3.39和2.99、2.03和2.20、3.23和3.48,P<0.05或P<0.01);而IL-6、IL-8、TNF-a、IgA则显著升高(t值分别为6.97和5.83、8.17和8.31、9.13和8.85、4.01和2.88,均P<0.01).菌苗治疗组经母牛分枝杆菌菌苗治疗后CD4+、CD4+/CD8+、NK细胞升高,与治疗前比较,差异有统计学意义(t值分别为3.09、2.07、3.17,P<0.05或P<0.01);而IL-6、IL-8、TNF-a、IgA则显著降低(t值分别为4.62、5.15、7.26、3.35,均P<0.01).常规治疗组治疗前、后各项指标差异均无统计学意义(P>0.05).随访1年中,菌苗治疗组患者人均年急性加重次数和住院次数与常规治疗组比较下降,差异均有统计学意义(t值分别为10.99和5.89,均P<0.01).结论 母牛分枝杆菌菌苗能改善COPD稳定期患者的细胞免疫功能,降低其急性加重及住院次数,具有较高的临床应用价值.     

慢性阻塞性肺疾病患者纤毛摆动时间的研究

目的 探讨慢性阻塞性肺疾病(COPD)患者纤毛摆动时间和肺功能的关系,及黏液纤毛清除功能(MCC)在COPD发病中的作用.方法 手术切除的支气管组织30例,分为COPD组、吸烟组及对照组,每组10例,光镜下观察活性纤毛细胞的百分率及纤毛摆动时间,并计数各个时间点的摆动纤毛细胞的百分率,收集临床资料包括吸烟指数、体质量指...     

二甲双胍在高海拔地区糖尿病治疗中的安全性研究

高海拔地区初诊糖尿病患者61例,个体化治疗基础上全部加用二甲双胍1.5g/d治疗14 d.平均海拔2 260米和2 780米两地区(海拔差580米),氧分压相差3.5 mm Hg(1 mm Hg=0.133 kPa),初发糖尿病患者在治疗前血乳酸水平即高于正常值.高海拔地区中青年组及老年组治疗前血乳酸水平均较高[(3.90±0.85对3.65±0.70)、(4.67±0.80对3.69±0.78)mmol/L,均P＜0.05],中青年组糖尿病患者加用二甲双胍14 d后血乳酸水平未显著上升[(4.50±0.50和3.79±0.62)mmol/L,P＞0.05],老年组糖尿病患者加用二甲双胍14 d后血乳酸水平明显上升[(5.59±0.55和5.27±0.43)mmol/L,P＜0.05].因此,在高原地区的老年糖尿病患者应慎用或避免使用二甲双胍.     

慢性阻塞性肺疾病肺血管内皮细胞和肺泡上皮细胞凋亡的实验研究

目的 研究COPD患者及烟雾暴露COPD大鼠肺血管内皮细胞和肺泡上皮细胞凋亡的相互关系及其与肺功能和肺气肿的相关性.方法 采用烟雾暴露法建立COPD大鼠模型.分别采集COPD患者(13例)和对照患者(12例)及烟雾暴露80 d大鼠(11只)和对照大鼠(12只)的肺组织标本,用HE染色评估肺部病理改变,用平均内衬间隔(MLI)与平均肺泡数(MAN)评估大鼠肺气肿程度;用原位末端标记法对肺血管内皮细胞和肺泡上皮细胞凋亡进行定量检测,分析这两种细胞凋亡的相关性,及其与肺功能和肺气肿指标的相关性.正态分布的计量资料以x±s表示,采用两独立样本的t检验进行分析;非正态分布的计量资料以中位数(四分位间距)表示,采用两个独立样本比较的Wilcoxon秩和检验;采用Spearman秩相关进行相关分析.结果 COPD患者和大鼠均出现明显肺气肿病理改变.COPD患者肺血管内皮细胞和肺泡上皮细胞凋亡指数[(13.2±2.6)%和(28.9±3.1)%]明显高于对照患者[(5.6±1.5)%和(5.8±1.2)%].COPD患者肺泡上皮细胞凋亡指数[(28.9±3.1)%]明显高于肺血管内皮细胞凋亡指数[(13.2±2.6)%],二者间呈正相关(r=0.60,P<0.05);COPD患者肺血管内皮细胞和肺泡上皮细胞的凋亡指数与FEV.占预计值%呈负相关(r值分别为-0.83和-0.69,均P<0.05),与FEV1/FVC呈负相关(r值分别为-0.95和-0.71,均P<0.05),与残气容积/肺总量呈正相关(r值分别为0.93和0.70,均P<0.05).COPD大鼠肺血管内皮细胞和肺泡上皮细胞凋亡指数[(4.1±0.4)%和(10.0±1.0)%]明显高于对照大鼠[(0.2±0.1)%和(2.1±0.4)%],COPD大鼠肺组织MLI与肺泡上皮细胞凋亡指数呈正相关(r=0.59,P<0.05),MAN与肺泡上皮细胞凋亡指数呈负相关(r=-0.81,P<0.05).结论 COPD患者和大鼠肺内存在异常的细胞凋亡现象,其肺泡上皮细胞凋亡较肺血管内皮细胞凋亡更为明显,且二者呈正相关;COPD患者和大鼠肺血管内皮细胞和肺泡上皮细胞的凋亡与其肺功能及肺气肿的改变明显相关,推测肺血管内皮细胞和肺泡上皮细胞凋亡可能参与COPD的发病过程.     

硼替佐米对脑缺血再灌注损伤保护作用的研究

目的 观察硼替佐米对脑缺血再灌注后炎性反应及细胞凋亡的影响,探讨其脑保护作用机制.方法 将15只大鼠随机分为假手术组、生理盐水组、硼替佐米组各5只.脑缺血2 h再灌注即刻给予硼替佐米0.2 mg/kg尾静脉注射,对照组以等量生理盐水尾静脉注射,再灌注后24 b取材.免疫组化法测组织核因子(NF)-κBp65、白细胞介素(IL)-1β;缺口末端标记法检测神经细胞凋亡.结果假手术组偶见NF-κBp65、IL-1β免疫反应阳性细胞[分别为(1.21±0.16)个/400倍、(11.56±0.99)个/400倍],凋亡细胞亦少见[(2.88±0.27)个/400倍],生理盐水组与硼替佐米组均可见大量NF-κBp65、IL-1β免疫反应阳性细胞及凋亡细胞[分别为(56.28±1.95)个/400倍与(29.76±2.53)个/400倍、(47.64±2.06)个/400倍与(29.6±1.61)个/400倍、(51.05±4.23)个/400倍与(33.44±2.06)个/400倍],差异有统计学意义(均P＜0.05).结论 蛋白酶体抑制剂硼替佐米可通过减少NF-κB的激活,抑制炎性反应,减少细胞凋亡,对缺血脑组织有保护作用.     

细胞色素C氧化酶在慢性阻塞性肺疾病中的表达及其与肺血管内皮细胞凋亡的关系

目的 观察COPD患者肺组织中细胞色素C氧化酶(COX)和血管内皮细胞凋亡的变化,探讨COPD的发病机制.方法 2007年9月至2008年5月在中南大学湘雅二医院行肺切除术的周围型肺癌患者20例,参照COPD的诊断标准和患者的吸烟状况分为对照组(7例)、吸烟非COPD组(7例)和吸烟并COPD组(6例),取远离病变的肺组织.采用脱氧核糖核苷酸末端转移酶介导的缺口末端标记法检测肺血管内皮细胞凋亡,采用分光光度法、半定量逆转录PCR法和Western blot法分别检测肺组织中COX活性、COXⅡ亚基(COXⅡ)mRNA和蛋白表达,免疫组织化学法检测肺血管内皮细胞COXⅡ蛋白的分布和表达.多组间比较采用单因素方差分析,用LSD-t检验进行两两比较.结果 吸烟并COPD组肺血管内皮细胞凋亡指数为(13.8±1.9)%,明显高于吸烟非COPD组[(9.6±0.8)%]和对照组[(5.9±1.0)%];吸烟并COPD组肺组织中COX活性为(4.4±0.7)×105U/kg,COXⅡmRNA相对表达量为0.76±0.17,肺血管内皮细胞中COXⅡ蛋白相对表达量为14.5±1.6,明显低于吸烟非COPD组[(6.0±0.6)×105 U/kg、0.81±0.15和18.6±2.1]和对照组[(7.6±0.4)× 105U/kg、0.86±0.20和23.8±3.4],差异均有统计学意义(t值为-13.66～13.27,均P＜0.05);COX活性与FEV1占预计值%和FEV1/FVC呈显著正相关(r值分别为0.84和0.91,均P＜0.01),与吸烟指数呈显著负相关(r=-0.78,P＜0.01);肺血管内皮细胞凋亡与COXⅡ蛋白表达呈显著负相关(r=-0.75,P＜0.01).结论 COPD患者的肺血管内皮细胞凋亡增加,COX表达和活性下降,且均与吸烟有相关性.COX可能参与介导COPD肺血管内皮细胞凋亡过程.     

老年慢性阻塞性肺病患者窦性心率震荡及心率变异性的研究

目的 探讨慢性阻塞性肺病(COPD)患者窦性心率震荡(HRT)及心率变异性(HRV)变化及临床意义.方法 临床诊断为COPD老年患者59例,选择同期健康老年人30例为对照组.24 h动态心电图检测各组HRT参数:震荡初始(TO)和震荡斜率(TS)及HRV各项指标,肺功能检测,同时超声心动图测量左心室射血分数(LVEF)、右心房内径(RAD)、右心室内径(RVD)、右心室壁厚度(RVWT)等指标;组间比较且对HRT与HRV指标进行相关性分析.结果 与对照组比较,COPD患者TO值显著增高[(-0.2±1.1)%与(-3.8±2.8)%,t=6.830,P＜0.01],TS值显著下降[(7.0±3.6)与(11.7±6.1)ms/RR,t=3.866,P＜0.01];HRV指标正常R-R间期的标准差(SDNN)、正常R-R间期的标准差的平均值(SDNNI)、SDNNI的标准差(SDANN)、相邻R-R间期之差的均方根值(rMSSD)和正常R-R间期标准差≥50 ms的百分数(PNN50)增加,且随肺动脉压力的增高而恶化.TO与SDANN,rMSDD呈负相关(r=-0.369,P＜0.05;r=-0.472,P＜0.01),TS和SDNN,SDANN,PNN50呈正相关(P＜0.05),与rMSDD无相关性(P＞0.05).结论 COPD患者HRT现象减弱,HRT和HRV变化随患者肺动脉压力的增加而恶化,联合检测对评价COPD患者自主神经功能状态及预后有较高临床价值.

Abstract：

Objective To explore the clinical significance of sinus heart rate turbulence (HRT)and heart rate variability (HRV) in patients with chronic obstructive pulmonary disease(COPD).Methods The 59 moderate to severe COPD patients and 30 healthy subjects were enrolled in this study. The 24-hour holter monitor was used to screen the HRT onset (TO), turbulence slope (TS)and HRV. Pulmonary function tests and echocardiographic examination were performed for measuring left ventricular ejection fraction (LVEF), right atrial dimension (RAD), right ventricular dimension (RVD), right ventricular wall thickness (RVWT). Then all the parameters were compared between NC group and COPD group, and the relationship between HRT and HRV was investigated. Results Compared with control group, TO was significantly increased [(-0.2±1.1) % vs.(-3.8±2.8) %, t=6. 830,P＜0.01] and TS was decreased [(7.0±3.6) ms/RR vs. (11.7±6.1) ms/RR, t =3. 866, P＜0.01] in COPD group. In time domain HRV parameters, normal RR intervallerinin standart deviation(SDNN), standard deviation of normal-to-normal beats index (SDNNi), standard deviation of the averages of normal sinus to normal sinus (SDANN), mean squared differences of the successive RR intervals (rMSDD), fraction of consecutive normal sinus intervals that differ by more than 50 ms (PNN50) were significantly lower in COPD group than in control group(P＜0. 05). TO was negatively correlated with SDANN and rMSDD (r=-0. 369, P＜0. 05; r=-0.472, P＜0.01).TS was positively correlated with SDNN, SDANN and PNN50 (all P＜0.05), but had no correlation with rMSDD (P＞0. 05). Conclusions HRT and HRV are dramatically blunted in COPD patients.Combination of HRV and prognosis. and HRT may be simple and elegant ways for evaluating cardiac autonomic functions.     

肠内营养对慢性阻塞性肺疾病模型大鼠的肠黏膜屏障的影响

目的：探讨肠内营养（EN）能否改善COPD的肠黏膜屏障。方法采用随机表法将35只大鼠分为空白对照组（K组，n＝10只）、COPD组＋EN组（n＝25只）。对COPD组＋EN组共25只大鼠进行COPD模型制备，将COPD组＋EN组共19只大鼠再随机分为COPD组和EN组2组，第29天开始给予EN组EN混悬液喂养14d ，COPD组和K照组继续给予普通饮食，2周后测定各组大鼠的肺功能，采血检测丙二醛、二胺氧化酶，无菌条件下取肠系膜淋巴结行细菌培养，取回肠组织用光学显微镜测量黏膜厚度、隐窝深度、绒毛高度，用透射电镜观察小肠黏膜上皮细胞的微绒毛和细胞间的链接复合体，用免疫组织化学检测回肠黏膜固有层IgA＋浆细胞数量。结果 COPD组与EN组回肠黏膜厚度与 K 组相比均降低（528.92±59.32 v s 465.89±62.72、528.92±59.32 v s 393.52±72.64），而COPD组降低更明显（465.89±62.72 v s 393.52±72.64），差异有统计学意义（P＜0.05）。绒毛高度和隐窝深度在 K 组和 EN 组间比较（331.54±34.26 vs 308.76±20.08、143.17±24.29 vs 136.77±16.96），差异无统计学意义（P＞0.05），COPD组与EN组相比，绒毛高度和隐窝深度指标下降更明显（308.76±20.08 v s 249.93±33.42、114.50±11.44 v s 136.77±16.96），差异有统计学意义（P＜0.05）。EN组与K组细菌移位率差异无统计学意义（P＞0.05）， EN组比COPD发生细菌移位的概率低（ P＜0.05）。血清丙二醛及二胺氧化酶浓度在 K组与EN组之间比较（0.86±0.43vs1.318±0.29、7.46±1.78vs8.02±2.45），差异无统计学意义（P ＞0.05），而COPD组的血清丙二醛、二胺氧化酶浓度明显比EN组的浓度高（1.829±0.57vs1.318±0.29、11.10±3.23vs 8.02±2.45），差异有统计学意义（ P＜0.05）。EN组与K组相比肠黏膜固有层IgA＋浆细胞数均有所下降（40.50±3.95vs51.80±4.87），而COPD组下降更明显（34.11±6.29vs40.50±3.95），差异有统计学意义（P＜0.05）。结论 COPD会导致肠黏膜屏障的损伤， EN可以改善COPD的肠黏膜屏障。     

雷公藤甲素对2型糖尿病大鼠肾组织足细胞Nephrin、Podocin蛋白表达的影响及机制

目的 观察雷公藤甲素对2型糖尿病(T2DM)大鼠肾组织足细胞Nephrin、Podocin蛋白表达的影响及其机制探讨.方法 SPF级雄性8周龄Wistar大鼠50只,体重(200±20)g,按随机数字表法分为2组:对照组10只、模型组40只.模型组高脂高糖喂养8周联合小剂量链脲佐菌素(STZ)30mg/kg建立T2DM模型鼠,正常组大鼠喂养常规饲料.将造模成功的T2DM大鼠(28只)随机分成2组:糖尿病组(14只)和雷公藤甲素治疗组(14只),雷公藤甲素治疗8周后,检测大鼠生化指标、肾重/体重和尿白蛋白.利用光镜、电镜观察肾脏病理改变.采用实时定量PCR、免疫组化及Western blot法检测肾组织Nephrin及Podocin蛋白、骨桥蛋白、转化生长因子-β1(TGF-β1)及单核/巨噬细胞表面特异性标志抗原(ED-1)的mRNA和蛋白表达分布.采用单因素方差分析法进行统计学分析.结果 糖尿病大鼠尿白蛋白明显高于对照组(F=181.51,P＜0.01),治疗组尿白蛋白明显低于DM组(F=181.51,P＜0.01).与对照组比较,糖尿病大鼠肾组织Nephrin(F=36.82,P＜0.05)和Podocin(F=32.57,P＜0.05)蛋白表达下调,Nephrin(F=88.45,P＜0.01)、Podocin(F=55.43,P＜0.01)mRNA表达下调;雷公藤甲素组干预8周后Nephrin[(1.82±0.06)和(1.63±0.06),P＜0.05]和Podocin[(1.80±0.06)和(1.60±0.06),P＜0.05]蛋白质表达上调,肾脏组织Nephrin[(0.73±0.12)和(0.19±0.08),P＜0.01]和Podocin[(0.60±0.12)和(0.26±0.07),P＜0.01]mRNA表达上调.与对照组比较,T2DM大鼠肾组织骨桥蛋白(F=40.06,P＜0.05)和TGF-β1的(F=28.84,P＜0.05)蛋白质表达上调,肾组织骨桥蛋白(F=177.46,P＜0.01)和TGF-β1(F=161.27,P＜0.01)mRNA表达上调,雷公藤甲素治疗后肾组织骨桥蛋白[(1.27±0.03)和(1.39±0.05),P＜0.05]和TGF-β1[(1.23±0.03)和(1.44±0.02),P＜0.05]的蛋白表达下调,肾组织骨桥蛋白[(2.05±0.16)和(3.26±0.26),P＜0.01]和TGF-β1[(4.0±0.70)和(6.5±0.71),P＜0.01]mRNA表达下调.结论 雷公藤甲素对T2DM大鼠足细胞有保护作用,其机制可能与其?     

携带人B型利钠肽基因重组腺病毒对慢性心力衰竭大鼠心功能的影响

目的 观察外源基因人B型利钠肽对慢性心力衰竭(心衰)大鼠心功能的影响.方法 30只入选心衰大鼠,随机分为携带人B型利钠肽基因重组腺病毒组(Ad-hBNP组)、重组空白腺病毒组(Ad-Track组)、生理盐水组(NS组),另设不予任何治疗的假手术组作为对照;分别经腹腔注射予以相应治疗,每周1次,共4周.4周后实验动物行超声心动图、血流动力学检测,酶联免疫吸附试验检测血清外源基因人B型利钠肽水平,全心质量指数检测.结果 间断Ad-hBNP治疗后,Ad-hBNP组心衰大鼠室间隔厚度、左室后壁厚度、左室舒张末径、左室收缩末径[(2.34±0.29)mm、(2.28±0.18)mm、(6.50±0.42)mm、(3.54±0.59)mm]显著低于Ad-Track组[(2.71±0.35)mm、(3.02±0.85)mm、(7.71±0.83)mm、(4.72±0.80)mm,均为P＜0.05]和NS组[(2.78±0.23)mm、(2.83±0.53)mm、(7.34±0.97)mm、(4.55±0.77)mm,均为P＜0.05],而左室射血分数、左室短轴缩短率[(79.27±7.01)%、(43.38±6.73)%]显著高于Ad-Track组[(70.85±4.81)%、(35.72±3.68)%,均为P＜0.01]和NS组[(69.67±6.90)%、(34.91±5.10)%,均为P＜0.01].Ad-hBNP组与Ad-Track组和NS组比较:心率显著降低,左室收缩压显著升高[为(131.79±15.76)mm Hg(1 mm Hg=0.133 kPa)、(112.99±32.35)mm Hg、(117.13±15.26)mm Hg],左室内压最大上升速率显著升高[分别为(5037.20±430.41)mm Hg/s、(4217.40±1354.15)mm Hg/s、(4310.50±1293.97)mm Hg/s;P＜0.05];左室内压最大下降速率显著升高[分别为(-4382.00±1304.79)mm Hg/s、(-3725.00±791.34)mm Hg/s、(-3890.00±1043.73)mm Hg/s,均为P＜0.05];左室舒张末压降低[分别为(-4.24±4.00)mm Hg、(21.99±6.80)mm Hg、(18.00±12.25)mm Hg,均为P＜0.01];心脏质量及全心质量指数均降低.结论 间断给予Ad-hBNP能够有效地改善心衰大鼠心脏结构和功能.

Abstract：

Objective To evaluate the therapeutic effect of hBNP on rats with chronic heart failure (CHF). Methods Thirty CHF rats defined by echocardiography at 12 weeks post abdominal aortic constriction were randomly divided into Ad-hBNP group (2. 5 × 1010 VP/ml NS Ad-hBNP 1 ml/week ×4,n = 14), Ad-Track group ( n = 8 ), placebo group ( NS, n = 8 ), 10 sham-operated rats served as control group. After 4 weeks treatment, cardiac function was evaluated by echocardiography and hemodynamic measurements. Heart weight (HW) and HW/body weight (BW) ratio were determined. Results IVS,LVPW, LVEDD and LVESD were significantly reduced in the Ad-hBNP group [(2. 34 ±0. 29)mm, (2. 28 ± 0. 18)mm, (6. 50 ±0. 42)mm, (3.54 ±0. 59) mm] than those in the Ad-Track group[(2. 71 ±0. 35) mm,(3.02 ±0.85)mm, (7.71 ±0.83)mm, (4.72 ±0.80)mm] and in the NS group [(2.78 ±0.23)mm,(2. 83 ± 0. 53 ) mm, (7. 34 ± 0. 97 ) mm, (4. 55 ± 0. 77 ) mm, all P ＜ 0. 05]. The LVEF and LVFS of the Ad-hBNP group [(79. 27 ±7.01 )%, (43.38 ±6. 73)%] were significantly higher than in the Ad-Track group[(70.85±4.81)%, (35.72 ±3.68)%] and in the NS group[(69.67 ±6.90)%, (34.91 ±5.10)%, all P ＜0. 01]. HR[(417.48 ±32. 57) beats/min, (446. 85 ±61.49) beats/min, P ＜0. 05;(440. 83 ±32. 18) beats/min , P ＜0. 05], LVEDP[( - 4. 24 ±4. 00) mm Hg( 1 mm Hg =0. 133 kPa);(21.99 ±6. 80) mm Hg, P ＜0. 01; ( 18.00 ± 12. 25)mm Hg, P＜0. 01] were significantly decreased and while LVSP[(131.79 ±15.76) mm Hg; (112.99 ±32.35) mm Hg, P＜0.05; (117.13 ±15.26)mmHg], +dP/dtmax[(5037.20 ±430.41) mm Hg/s; (4217.40 ± 1354. 15)mm Hg/s, P ＜0.05;(4310. 50 ± 1293.97 ) mm Hg/s, P ＜ 0. 05] and - dP/dtmax [( - 4382. 00 ± 1304. 79 ) mm Hg/s;(-3725.00±791.34) mm Hg/s, P ＜ 0.05; ( - 3890.00 ± 1043.73) mm Hg/s, P ＜ 0.05] were significantly increased in Ad-hBNP group than in Ad-Track group and NS group ( all P ＜ 0. 05 ). HW and HW/BW were also decreased in Ad-hBNP group than in the Ad-Track group and the NS group. Conclusion Exogenous hBNP improved the cardiac function and attenuated remodeling in CHF rats.