DETECTION OF BCL-2 GENE MAJOR BREAKPOINT REGION REARRANGEMENT IN HUMAN B-CELL LYMPHOMAS

Objective To investigate the frequency of t(14; 18) in different subtypes of B-cell lymphomas and the ability or the polymerase chain reaction(PCR) to detect this rearrangement in frozen samples. Methods 1o7 cases of B-cell lymphomas were studied uslng DNA extracted from rresh-frozen tissues. The DNA samples were amplified by PCR for bcl-2 MBR/JH. The products of bcl-2/JH rearrangement were hybridized with an internal olignucleotide probe or bcl-2 MBR. Results The rearranged bcl-2MBR/JH gene was detected in 13 of the 25(52. o% ) follicular center lymphomas, according to REAL classification: 8 of 11 (72. 7%) grade 1, 2 of 5(40. 0%) grade I, and 3 of 90 (33. 3%) grade, 17 of 82(2o. 8%) cases or difruse large B-cell lymphomas were found to have detectable bel-2 MBR/J. rearrangement- Conclusion The rrequency or bcl-2 MBR/JH rearrangement in diffuse large B-cell lymphomas is significantly lower than those in follicular center lympkomas(X2= 9. 28, P <o. oo5), suggesting that bcl2/JH rearrangements occur mainly in follicular center lymphomas. in addition, the result of reconstruction experiments suggest that amplification or bcl-2 MBR/JH rearrangements by PCR is both sensitive and specific for detection of t (14; 18 ) translocation.

The Expression and Significance of CyclinD2 and Bcl-2 in Diffuse Large B-cell Lymphoma

Objective:To study the expression and significance of cyclinD2 and Bcl-2 in diffuse large B-cell lymphoma.Methods:This project used immunohistochemical methods to detect the expression of cyclinD2 and Bcl-2 in 120 cases of diffuse large B-cell lymphoma and 80 cases of reactive hyperplasia of lymphoid tissue.The materials were collected from the hospital from January 2018 to 2020.In March 2015,120 patients had lymphoma tissue removed during the month of surgery.The postoperative pathological diagnosis was DLBCL.Another 80 cases of lymphoid hyperplasia(RLH)tissues were selected as controls.Results:CyclinD2 and BCL-2 expression were not statistically different in patients with diffuse large B-cell lymphoma in different ages,genders,locations,tissue types,and degree of differentiation;but statistically significant in different Ann Arbor stages,immunotypes,IPI index and first treatment efficacy.Conclusion:This research not only has important theoretical value,but also important economic value and social significance.

湿润烧伤膏对糖尿病足创面组织中MMP-2、MMP-9、Bcl-2、Bax水平的影响

目的探讨湿润烧伤膏(MEBO)对糖尿病足创面组织中基质金属蛋白酶(MMP)-2、MMP-9、B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)水平的影响。方法选取2020年5月至2022年5月郑州大学附属郑州中心医院收治的55例糖尿病足患者作为研究对象,按照不同治疗方法将其分为MEBO组(35例)和VSD组(20例),MEBO组患者局部创面采用MEBO治疗,VSD组患者局部创面采用负压封闭引流(VSD)治疗,对比观察两组患者创面面积,创面组织中MMP-2、MMP-9、Bcl-2、Bax水平及临床疗效。结果治疗第7、21天,MEBO组患者创面面积均明显小于VSD组(t=2.719、5.268,P=0.009、P<0.001),创面组织中MMP-2、MMP-9、Bax水平均明显低于VSD组(MMP-2:t=2.138、2.202,P=0.037、0.032;MMP-9:t=2.129、2.476,P=0.038、0.017;Bax:t=3.623、3.038,P=0.001、0.004),Bcl-2水平均明显高于VSD组(t=2.040、3.054,P=0.046、0.004);治疗21 d后,MEBO组患者中显效23例、有效10例、无效2例,明显优于VSD组患者的显效8例、有效7例、无效5例(Z=-2.126,P=0.033)。结论MEBO可通过降低糖尿病足创面组织中MMP-2、MMP-9、Bax水平以及提高Bcl-2水平促进创面愈合。

Prognostic value of BCL2 and TP53 genetic alterations for diffuse large B-cell lymphoma patients treated with R-CHOP

Objective:Limited data about the prognostic significance of BCL2 mutations and BCL2 copy number variations in diffuse large B-cell lymphoma(DLBCL)are available.This study aimed to comprehensively describe BCL2 genetic alterations in DLBCL patients,and examine correlation of BCL2,TP53 and other genetic alterations with outcomes in patients treated with R-CHOP.Methods:Probe capture-based high-resolution sequencing was performed on 191 patients diagnosed with de novo DLBCL.MYC,BCL2,and BCL6 protein expressions were detected by immunohistochemistry.Results:The presence of BCL2 alterations significantly correlated with poor progression-free survival(PFS)(5-year PFS:13.7%vs.40.8%;P=0.003)and overall survival(OS)(5-year OS:34.0%vs.70.9%;P=0.036).Importantly,patients who harbored BCL2 gain/amplifications(BCL2GA/AMP)also had a remarkably inferior 5-year PFS(11.1%vs.38.3%;P<0.001)and OS(22.1%vs.69.6%;P=0.009).In contrast,neither BCL2 mutations nor BCL2 translocations were significantly prognostic for survival.Multivariable analyses showed that the presence of BCL2 alterations,especially BCL2GA/AMP,TP53 mutations,and International Prognostic Index(IPI)were significantly associated with inferior PFS and OS.Novel prognostic models for OS were constructed based on 3 risk factors,including BCL2 alterations(Model 1)or BCL2GA/AMP(Model 2),TP53 mutations,and IPI,to stratify patients into 4 risk groups with different survival outcomes.Conclusions:This study showed that DLBCL patients treated with R-CHOP,BCL2 alterations,especially BCL2GA/AMP and TP53 mutations were significantly associated with inferior outcomes,which were independent of the IPI.The novel prognostic models we proposed predicted outcomes for DLBCL patients treated with R-CHOP,but further validation of the prognostic models is still warranted.

Bcl-2 GENE REARRANGEMENT DETERMINED BY PCR AS AMEAN TO DETECT MINIMAL RESIDUAL DISEASE INMALIGNANT LYMPHOMAS

Objective: To develop a sensitive method to detect minimal residual disease and to elucidate the significance of bcl-2 gene rearrangement in diagnosis and treatment of malignant lymphoma. Methods: Using polymerase chain reaction (PCR) to detect bcl-2 gene rearrangement and using serial dilution method to define the sensitivity of PCR. Results: In 9 different malignant lymphoma cell lines, Su-DHL-4 and Su-DHL-6 were shown bcl-2(MBR)/JH rearrangement, the sensitivity of PCR was 1:105. In 16 patients with follicular lymphoma, the peripheral blood and bone marrow were PCR positive in 4 cases both at initial diagnosis and after complete remission. Conclusion: Detection of bcl-2 gene rearrangement by PCR provides a sensitive and specific assay of minimal residual disease. It is helpful to improve staging of disease, prognosis and evaluation of the treatment results.

Total saponins in Rubus parvifolius L.induce lymphoma cells apoptosis through upregulated Bax/Fas and downregulated Bcl-2 in vivo and in vitro

Purpose:To investigate the effect of total saponins of Rubus parvifolius L.(TSRP)on lymphoma Raji cells and further discuss its mechanism.Methods:The model of nude mice bearing Raji cells was established,the volume,weight and inhibition rate of the transplanted tumor were analyzed and compared after different concentrations of TSRP treatment.Cell apoptosis and expression of Bcl-2,Bax,Fas proteins were detected by TUNEL and immunohistochemiscal method respectively.Effects of TSRP on cell proliferation were tested with MTT assay in vitro.Cell apoptosis and expression of Caspase-9,Caspase-3,Bcl-2,Bax and Fas proteins were tested with DAPI staining and Western blot.Results:TSRP significantly reduced the volume and tumor weight of Raji subcutaneous transplanted tumor and induced the apoptosis of Raji cells in vivo.The tumor inhibition rate of high-dose(100 mg/kg)TSRP is 90.84%.The TUNEL test results show that the fluorescence intensity of the tumor issue treated with TSRP is significantly improved.Compared with the control group,the fluorescence intensity of high-concentration TSRP is 82.43 ± 7.81,which is significantly different(P<0.001).The results of immunohistochemistry test showed that the Bcl-2 expression of Raji cell treated with TSRP is obviously reduced,and Bax expression is obviously increased.Meanwhile,compared with that of control group,Fas expression is obviously reduced.MTT assay showed that TSRP can significantly inhibit proliferation of Raji cells with dose dependence.The inhibition rate of 400 μg/mL TSRP is 53.46 ± 4.90%(P<0.001).DAPI staining results showed that TSRP can significantly induce cell apoptosis.According to Western blot results,it is found that TSRP can significantly inhibit activity of Bcl-2 and increase Bax expression,and TSRP can also inhibit Fas expression.Meanwhile,expression of Caspase-9 and Caspase-3 is also increased.Conclusion:TSRP could inhibit the proliferation of lymphoma via induction of apoptosis in a time and dose-dependent manner.Apoptotic signaling induced by TSRP was characterized by up-regulating Bax,Fas and Caspase-8 protein expression,and down-regulating of Bcl-2 protein expression.

Detection of apoptotic cells and immunohistochemical study of bcl-2 and p53 gene protein in primary gastric mucosa-associated lymphoid tissue (MALT) lymphoma

To identify the apoptotic cells in gastric MALT lymphoma and its relationship between bcl-2 and p53 gene expression. Methods: TdT-mediated dUTP biotin Nick End labeling (TUNEL) and immuno-histochemistry ABC method were used to display apoptotic cells and the gene protein expression of bcl-2 and p53 independently. Results: Apoptotic indices (AI) in high-grade MALT lymphomas were significantly higher than in mixed-grade group and low-grade group (P<0.05). Bcl-2 was expressed in 83% of low-grade tumors, 61.6% of the median-grade tumors and 43.7% of high-grade tumors. An inverse correlation was observed between the expression of bcl-2 and apoptotic indices. Only 27 cases were p53 positive. The frequency of p53 positivity was significantly increased as the histologic grade advanced (P<0.05). There was also an inverse correlation between the expression of bcl-2 and p53. Conclusion: Apoptosis may be important in tumors development and transmission. p53 and bcl-2 were important regulatory genes of apoptosis and may be associated with transformation from low- grade to high-grade lymphomas.

CHOP-like Regimen in Combination with Rituximab and Peginterferon Alpha-2b in Newly-diagnosed Diffuse Large B-cell Non-Hodgkin＇s Lymphoma： Experience in a Chinese Center

OBJECTIVE To evaluate the efficacy of rituximab combined with CHOP-like regimen with or without IFN in patients newly diagnosed diffuse large B-cell Non-Hodgkin＇s lymphoma （DLBCL）.METHODS From January 2003 to July 2008, 51 patients received CHOP-like chemotherapy （cyclophosphamide 750 mg/m2, epirubicin 80 mg/m2, vindesine 2.8 mg/m2 on day 1, and prednisolone 100 mg/day on day 1 to day 5）. Thirty-one patients received CHOPR-like treatment （rituximab 375 mg/m2 1 day before CHOP-like chemotherapy）. Twenty patients received CHOP-like regimen in combination with peginterferon （pegIFN） （1μg/kg on day 5） and rituximab （on day 6）.RESULTS -The CR （complete remission） rate in the CHOPR-like （with or without pegIFN） group and in the CHOP-like group was 78.4% and 45.1% （P = 0.005）, respectively. The estimated mean time of overall survival （OS） in the CHOPR-like group and CHOP- like group was 58.7 ± 2.8 and 36.4 ±3.4 months, respectively （P = 0.002）. The rates of CR and OR （overall remission） in CHOPR- like with IFN arm were 85.0% and 95.0%, and the rates of those in CHOPR-like without IFN arm were 74.2% and 87.0% （P 〉 0.05）. The estimated mean time of 4-year-PFS （progression- free survival） in CHOPR-like with IFN arm and in CHOPR-like without IFN arm was 62.9 ±3.0 months and 51.0 ± 4.6 months （P = 0.092）, respectively. In the CHOPR-like with IFN arm, no patient relapsed after achieving CR, while the estimated rate of 4-year- DFS （disease-free survival） in the patients who reached CR in the CHOPR-like without IFN arm was （63.4 ± 19.3）% （P = 0.061）. CONCLUSION Rituximab combined with CHOP-like chemotherapy improved the prognosis of DLBCL patients. The IFN may help to improve the quality and duration of response of DLBCL patients treated with rituximab and CHOP-like regimen.

Bcl-2 Small Interfering RNA Inhibits the Growth of Human Lymphoma Transplanted Subcutaneously in Nude Mice

OBJECTIVE To investigate whether small hairpin RNA (shRNA)targeting Bcl-2 mRNA could inhibit the growth of lymphomatransplanted subcutaneously in nude mice.METHODS Recombinant Bcl-2 shRNA expression vector withgreen fluorescence protein (GFP) gene was constructed andpreserved in our lab. We evaluated the antitumor effect of the Bcl-2shRNA in vivo which was the model of nude mice bearing Rajicells xenografts. Human Raji cells were injected subcutaneouslyinto nude mice to establish lymphoma models. When thediameters of tumor were above 0.5 cm after Raji cells injection,the mice bearing tumor were randomly divided into four groups:saline control group, negative shRNA group, plasmid vectorgroup, Bcl-2 shRNA group. The polyethylenimine (PEI) was usedto transfect shRNA into tumor. The mixed PEI and shRNA wasinjected into tumors. The growth and size of tumor were observed.Tissue was stained by H&E for its pathological morphology. Theexpression of Bcl-2 mRNA in the tumor mass was detected byreverse transcription polymerase chain reaction (RT-PCR).RESULTS A significant difference in median tumor weightwas observed in mice treated with Bcl-2 shRNA, compared withthose in the groups of negative shRNA or plasmid vector or salinesolution (P< 0.05). Pathological evaluation was completed in allexcised tumors from nude mice bearing Raji cells xenografts.The tumor tissue of the mice treated with Bcl-2 shRNA showedapoptosis, serious necrosis of the cells and inflammatory cellsinfiltration. There was no change in the morphology of cellsamong negative shRNA, plasmid vector and saline solution group.In the group of the Bcl-2 shRNA, the expression levels of Bcl-2mRNA of the tumor tissue were effectively inhibited (P < 0.05).CONCLUSION The shRNA targeting at the Bcl-2 mRNAcould inhibit the growth of human lymphoma transplantedsubcutaneously in nude mice.

Effect of rituximab combined with CVAD regimen on serum VEGF and β2-MG levels in patients with primary gastrointestinal B-cell lymphoma

Objective:To investigate the clinical effect of rituximab combined with CVAD regimen in patients with primary gastrointestinal B-cell lymphoma and serum vascular endothelial growth factor (VEGF) andβ2 microglobulin (β2-MG) The impact of the level.Methods:Eighty-four patients with primary gastrointestinal B-cell lymphoma treated from May 2014 to December 2015 were enrolled. Based on the random number table, all the patients were divided into a control group (n=42) and an observation group (n=42). The control group was treated with CVAD. The observation group was treated with rituximab on the basis of the control group. The effect of the patients was evaluated after 3 courses of treatment. The patients were followed up for 3 years after treatment. US RECIST 1.1 was used to evaluate the short-term efficacy on the patients;VEGF, TNF receptor-associated factor 6 (TRAF6) and B-cell lymphoma factor-6 (Bcl-6) levels were measured by enzyme-linked immunosorbent assay;β2-MG level test was implemented to compare the short-term efficacy, biochemical indicators, incidence of toxic side effects and long-term survival rate of the two groups. Results: The short-term efficacy rate of the observation group was 76.19%, which was higher than that of the control group (50.00%) (P<0.05). The levels of VEGF, TRAF6, Bcl-6, andβ2-MG were lower in the observation group after 3 courses of treatment than that in the control group (P<0.05);there was no significant difference in the incidence of neutropenia, gastrointestinal reactions, sepsis, infection, infusion-related reactions and cardiovascular events between the observation group and the control group (P>0.05);The 1-year long-term survival rate after treatment was not statistically significant (P>0.05). The long-term survival rate of the observation group was higher than that of the control group at 2 and 3 years after treatment (P<0.05).Conclusion: The combination of rituximab and CVAD in patients with primary gastrointestinal B-cell lymphoma can improve short-term efficacy, lower VEGF andβ2-MG levels, and lower incidence of side effects. It can improve the long-term survival rate of patients and is worthy of promotion and application.

高血脂大鼠脑缺血再灌注损伤后Bcl-2和Bax的表达变化及灯盏乙素的影响

目的观察高血脂大鼠缺血侧大脑皮质额叶内B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)的表达变化,探讨灯盏乙素对高血脂大鼠脑缺血再灌注损伤的保护作用。方法大鼠建立高血脂模型后,线栓法阻塞大脑中动脉建立脑缺血再灌注损伤模型,HE染色观察大脑皮质额叶的病理变化,免疫组织化学和免疫印迹法观察Bcl-2、Bax蛋白的表达变化,同时结合神经行为学评分和2,3,5-氯化三苯基四氮唑(TTC)染色观察大鼠的神经行为及脑梗死灶体积的变化。结果与假手术组相比,生理盐水组大鼠脑组织损伤加重,Bcl-2的表达明显减少,Bax的表达明显增高,同时大鼠的神经行为学评分和脑组织梗死灶体积明显增加(P<0. 05)。与生理盐水组相比,灯盏乙素治疗组脑组织损伤减轻,Bcl-2的表达明显增多,Bax的表达明显减少,同时大鼠的神经行为学评分和脑组织梗死灶体积明显降低(P<0. 05)。结论灯盏乙素对高血脂大鼠脑缺血再灌注损伤的保护作用可能是通过促进Bcl-2的表达,抑制Bax的表达实现的。

昆母汤对类风湿关节炎成纤维样滑膜细胞bax、bcl-2的影响

目的观察昆母汤醇提液(KMT)对人类风湿关节炎成纤维样滑膜细胞(RA-FLS)B淋巴细胞瘤-2基因(bcl-2)、bcl-2相关X蛋白(bax)m RNA及蛋白的影响,探讨昆母汤治疗RA的部分作用机制。方法滑膜组织取自行关节置换术或关节镜检查术的8例活动性类风湿关节炎(RA)患者,分离出FLS并进行原代培养,用逆转录-聚合酶链反应(RT-PCR)观察bax、bcl-2 m RNA表达的变化,采用酶联免疫吸附测定(ELISA)法检测bax、bcl-2蛋白含量的变化。结果与肿瘤坏死因子-α(TNF-α)组比较,KMT高剂量组可以明显促进bax m RNA表达,明显抑制bcl-2 m RNA表达,且bax m RNA/bcl-2 m RNA比值明显提高(P<0.05,P<0.01);KMT中、高剂量组可以提高bax蛋白含量,各剂量组均可降低bcl-2蛋白含量,bax/bcl-2的蛋白比值明显提高(P<0.05,P<0.01),且有一定的剂量依赖性。结论昆母汤可能通过抑制bcl-2的表达,同时促进bax的表达,调节bax/bcl-2比值来发挥对RA-FLS的促凋亡作用,具有治疗RA的潜在价值。

针刺介入时机对缺血性中风大鼠神经功能评分和Bcl-2、Bax表达的影响

目的研究不同时间点介入针刺治疗对缺血性中风大鼠神经功能评分和对缺血脑组织中B细胞淋巴瘤/白血病-2(B cell lymphoma/lewkmia-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)阳性表达的细胞数量的影响,探讨针刺治疗机制及介入时机。方法将200只雄性SD大鼠随机分为假手术组、模型组、2小时针刺组、72小时针刺组和168小时针刺组5组,除假手术组外,均采用线栓法制备大鼠局灶性永久性脑缺血模型,在术后1、3、7、14天,4个时间点采用Garcia复合评分法评价大鼠神经功能,用酶联免疫分析法探查缺血脑组织中的Bcl-2、Bax蛋白表达情况,并在显微镜下记录阳性细胞数目进行比较。结果针刺各组缺血性中风大鼠神经功能评分得到明显改善,优于模型组(P<0.001);2小时针刺组分别优于72小时、168小时组(P<0.01),差异有统计学意义。缺血性中风大鼠术后不同时间点脑组织中的Bcl-2,Bax表达均增高,1天达到高峰,随后下降,与假手术组比较有统计学意义(P<0.001,P<0.01)。术后2小时与模型组、72小时、168小时针刺组比较,可明显提高Bcl-2蛋白的表达、抑制Bax蛋白的表达,差异有统计学意义(P<0.001);72小时与168小时针刺组比较可明显提高Bcl-2蛋白的表达、抑制Bax蛋白的表达,差异有统计学意义(P<0.01)。结论针刺可明显提高缺血性中风大鼠的Garcia评分,改善其运动、感觉和协调功能;针刺可通过调节Bcl-2、Bax蛋白表达来抑制神经细胞凋亡,挽救缺血半暗带神经细胞的功能,而且在缺血性中风后越早介入针刺治疗疗效越好,存在一个最佳治疗时间窗。

甘氨鹅脱氧胆酸钠通过MAPK/ERK1/2介导Bcl-2在Ser70位点的磷酸化引起人肝细胞癌的抗药

B细胞淋巴瘤-2(Bcl-2)是一种重要的抗凋亡蛋白质,在多种人类肿瘤中普遍过表达。甘氨鹅脱氧胆酸钠(GCDA)与消化道肿瘤的发生发展密切相关,并能介导肝癌细胞对化疗药物的抵抗。本文旨在探讨在GCDA介导的人肝细胞癌(HCC)耐药性中Bcl-2的作用及其机制。本研究以肝癌细胞系为研究对象,Western印迹结果显示,Bcl-2在多种肝癌细胞系中均有表达。设计靶向Bcl-2的siRNA沉默HCC细胞系内源性Bcl-2的表达,发现Bcl-2沉默之后促进了化疗药物5-FU介导的HCC细胞凋亡。机制上,GCDA可介导Bcl-2在Ser70位点的磷酸化,而Ser70位点的磷酸化能够被PD98059(MAPK/ERK1/2抑制剂)所抑制。构建huBcl2-WT和huBcl2-S70A真核表达载体,脂质体转染HCC细胞系。用Annexin V-FITC/PI流式细胞术检测凋亡细胞。结果显示,huBcl2-WT过表达能抑制5-FU介导的凋亡,S70位点失活突变成A后,Bcl-2的过表达不能抑制5-FU介导的凋亡。本研究提示,GCDA通过MAPK/ERK1/2通路介导的Bcl-2 Ser70位点的磷酸化,在肝癌细胞的存活和抗药中发挥重要作用。抑制Bcl-2能够促进化疗药物5-FU介导的HCC细胞凋亡,该结果为治疗GCDA介导的耐药性肝癌提供新的思路。

内毒素血症幼年大鼠小肠Bcl-2、Bax mRNA的变化

目的:通过内毒素血症幼年大鼠小肠Bc l-2及Bax mRNA表达的变化,探讨小儿胃肠功能障碍的机制。方法:腹腔注射内毒素制备18 d龄大鼠内毒素血症模型,注射同等量生理盐水为对照组,分别取全部回肠,半定量RT-PCR法检测Bc l-2、Bax mRNA的表达。结果:正常对照和内毒素组小肠各时间点的Bc l-2 mRNA均为阴性表达,正常对照Bax mRNA表达较弱,内毒素组小肠各时间点的Bax mRNA表达均明显增强(P<0.01),死亡组与24 h之内其他各组的表达无显著差异,比72 h亚组表达明显增高(P<0.05)。内毒素组Bax与Bc l-2 mRNA表达的比值明显高于对照组。结论:内毒素通过调节基因Bc l-2、Bax mRNA表达的变化促进幼年大鼠小肠细胞凋亡可能为严重感染时胃肠功能障碍机制之一。

聚肌苷酸聚胞苷酸对高血脂大鼠脑缺血/再灌注后Bcl-2和Bax表达的影响及神经保护作用

目的观察聚肌苷酸聚胞苷酸(Poly-IC)处理对高血脂大鼠脑缺血/再灌注(I/R)损伤后Bcl-2和Bax表达的影响,同时检测脑梗死灶、血脑屏障通透性和行为损伤症状,探讨其对高血脂合并脑I/R损伤是否发挥神经保护作用。方法高血脂大鼠随机分为脑I/R组、Poly-IC预处理组、Poly-IC后处理组和假手术(Sham)组,每组20只。0~4分神经行为评分记录各组大鼠的神经行为表现,伊文思蓝染色法检测各组大鼠血脑屏障通透性,TTC染色法观察各组大鼠脑组织梗死情况,TUNEL染色法测神经细胞的凋亡情况,Western blotting观察Bcl-2和Bax相对表达水平。结果与sham组比较,I/R组神经行为损伤症状严重,评分明显升高(P<0.05)。而Poly-IC预处理和后处理组评分较I/R组均明显降低(P<0.05)。伊文思蓝染色结果显示,I/R组较sham组血脑屏障通透性明显增高(P<0.05),而Poly-IC预处理或后处理可明显改善血脑屏障通透性(P<0.05)。脑梗死体积检测结果显示,sham组未见梗死灶均匀红染,而I/R组脑组织可见白色梗死区。与I/R组比较,Poly-IC预处理和后处理组梗死灶体积均明显缩小(P<0.05)。I/R组大鼠大脑皮质内细胞凋亡指数较sham组显著升高(P<0.05),而Poly-IC预处理和后处理组大鼠大脑皮质内细胞凋亡指数较I/R组均明显降低(P<0.05)。与sham组比较,I/R组Bax表达明显升高(P<0.05),Bcl-2表达明显降低(P<0.05)。与I/R组比较,Poly-IC预处理和后处理组Bax表达均明显降低(P<0.05),Bcl-2表达明显升高(P<0.05)。结论Poly-IC预处理或后处理可改善神经行为损伤症状,减轻血脑屏障损害,缩小脑梗死体积,降低神经细胞凋亡指数,对高血脂大鼠脑I/R损伤起到神经保护作用;该保护作用可能与Bcl-2和Bax表达水平改变有关。

氦氖激光穴位照射对兔颞下颌骨关节病软骨细胞Bcl-2与Bax的影响

目的:通过研究氦氖激光对退变颞下颌关节细胞凋亡相关蛋白Bcl-2与Bax表达的影响,明确氦氖激光对颞下颌关节退变的保护作用机制,为临床运用氦氖激光治疗颞下颌关节骨关节病提供依据。方法:将骨骼发育成熟的健康成年雄性新西兰大白兔随机分为正常组、假模型组、模型组和治疗组,每组10只,组内又分为造模成功后第1d、11d两个取材时间点。参照常嘉等的造模方法,建立颞下颌关节骨关节病动物模型并行CT检查证实造模成功后纳入。治疗组采用氦氖激光照射患侧颊车、上关、下关、听宫、翳风穴,5min/穴/天,共10d;假模型组仅作关节囊切开后即缝合组织。运用q PCR检测蛋白多糖4(PRG-4)及骨形态发生蛋白2(BMP-2)m RNA水平,Western Blot方法检测各组织Bcl-2、Bax的蛋白表达水平。结果:正常组及假模型组不同时间点之间软骨基质基因表达无明显差异,建立TMJOA模型后,基因表达降低,基质逐渐降解,氦氖激光治疗10天后,PRG-4及BMP-2表达有所上升;凋亡相关蛋白Bcl-2、Bax的表达在正常组及假模型组之间无显著性差异,造模后抗凋亡蛋白Bcl-2表达减少,而促凋亡蛋白Bax增强;氦氖激光干预10天后Bcl-2的表达较前明显增强,而Bax的表达显著下降。结论:凋亡调节蛋白参与了氦氖激光穴位照射改善颞下颌关节软骨基质的合成与代谢。

PTEN与Bcl-2在鼻咽癌中的表达及意义

目的观察PTEN与Bcl-2在鼻咽癌(NPC)中的表达情况,探讨两者的表达及其联合表达在NPC发生、发展、侵袭和转移过程中的可能作用。方法采用免疫组织化学染色法,观察50例NPC石蜡标本和15例正常鼻咽黏膜标本中PTEN与Bcl-2的蛋白表达情况。结果正常鼻咽组织中,PTEN呈强阳性表达,而Bcl-2的表达仅局限在上皮层的基底细胞层。NPC中,PTEN阳性表达率为22.0%(11/50),Bcl-2为82.0%(41/50),与正常组比较,差异均有统计学意义。两者在NPC的表达强度无明显相关(r=-0.109,P=0.453)。结论在NPC组织中,PTEN的低表达或表达缺失和Bcl-2强阳性表达可能协同参与了NPC的发生,而在NPC的发展、浸润和转移过程中,两者未能表现出明显的相关性。

PCNA和Bcl-2蛋白在牙龈组织中的表达

给实验动物狗分别行金合金、不含Be的镍铬合金、含Be的镍铬合金全冠修复,建立实验动物模型.采用ICP-MS(电感耦合等离子体质谱仪)检测戴冠前后牙龈组织中金属离子的质量分数,同时应用免疫组化EnVision二步法检测PCNA(增殖细胞核抗原)、Bcl-2(B细胞白血病/淋巴瘤基因2)蛋白在牙龈组织中的表达.结果表明,镍铬合金全冠戴用后,牙龈组织PCNA,Bcl-2蛋白表达明显,而金合金全冠戴用后表达不明显.PCNA蛋白表达率与Ni,Cr,Si及Be离子质量分数正相关,而Bcl-2蛋白表达率与离子质量分数无明显的相关性.

血管内皮生长因子和bcl-2在肾母细胞瘤中的表达及意义

目的:探讨细胞凋亡抑制基因B-淋巴瘤细胞-2(B-cell lymphoma/leuke mia-2,bcl-2)和血管内皮生长因子(Vascular endothelial growth factor,VEGF)在肾母细胞瘤中的表达及其相互关系。方法:用免疫组织化学和原位杂交方法研究bcl-2和VEGF蛋白及mRNA在17例肾母细胞瘤和7例瘤旁正常肾组织的表达。结果:bcl-2蛋白在肾母细胞瘤组织中的表达(82.4%)高于瘤旁正常肾组织(28.5%);VEGF蛋白在肾母细胞瘤组织中的表达(58.8%)高于瘤旁正常肾组织(14.3%);bcl-2 mRNA和VEGF mRNA在肾母细胞瘤的表达率分别为88.2%和82.4%;bcl-2和VEGF的表达之间存在正相关关系(rs=0.633,P<0.01)。结论:bcl-2和VEGF在小儿肾母细胞瘤发生发展中起重要作用。