睾丸支持细胞对异基因T淋巴细胞的毒性作用

目的:分析睾丸支持细胞(塞尔托利氏细胞)对异基因T淋巴的细胞毒性作用。方法:实验于2006-01/07在解放军广州军区武汉总医院实验科完成。睾丸支持细胞取材于二三周龄SPF级Wistar雄性大鼠,由湖北省实验动物中心提供(许可证号:SCXK(鄂)-2003-005)。T淋巴细胞取材于体质量为200-250g的SPF级SD雄性大鼠脾脏,由湖北省实验动物中心提供(许可证号:SCXK(鄂)-2003-005)。将Wistar大鼠睾丸支持细胞与异基因SD大鼠T淋巴细胞共同培养。按睾丸支持细胞含量将实验分6组:对照组(0个睾丸支持细胞 DMEM/F12培养基100μL)、睾丸支持细胞1×103组(100μL)、睾丸支持细胞1×104组(100μL)、睾丸支持细胞1×105组(100μL)、睾丸支持细胞1×106组(100μL)及FasL单抗处理的睾丸支持细胞1×106组(经0.2μg抗FasL单克隆抗体封闭过的睾丸支持细胞1×106,100μL),每组均加1×105T淋巴细胞,6复孔培养48h,应用噻唑蓝比色法测定睾丸支持细胞对T淋巴细胞的毒性作用,用酶标仪于570nm处测各孔吸光度(A值),细胞毒性百分比(%)=(对照组A值-试验孔A值)/对照组A值×100%。结果:睾丸支持细胞1×103组、睾丸支持细胞1×104组、睾丸支持细胞1×105组及睾丸支持细胞1×106组对T淋巴细胞的细胞毒性百分比显著高于对照组、FasL单抗处理的睾丸支持细胞1×106组(8.75%,20.37%,65.07%,62.96%;0,3.85%,P<0.01);睾丸支持细胞1×103组-睾丸支持细胞1×105组随睾丸支持细胞数量增加,对T淋巴细胞的毒性作用也明显增加,当睾丸支持细胞数增加至1×105,杀伤作用的增加不再显著。FasL单抗处理的睾丸支持细胞1×106组与对照组相比差异无显著性意义(P>0.05)。结论:睾丸支持细胞对异基因T淋巴细胞有明显毒性作用,随着睾丸支持细胞增加,对异基因T淋巴细胞的抑制和杀伤作用也越大;但是当睾丸支持细胞达到一定数量后,其毒性作用不再增加;这种作用也可以被抗FasL单克隆抗体阻断。表明睾丸支持细胞FasL的表达是发挥免疫豁免作用的主要作用途径。     

Estrogen receptor and HER2/neu status affect epigenetic differences of tumor-related genes in primary breast tumors

Introduction

Estrogen receptor (ER)-positive breast cancers are considered prognostically more favorable than ER-negative tumors, whereas human epidermal growth factor receptor (HER)2/neu-positive breast cancers are associated with worse prognosis. The objective of the present study was to determine whether ER-positive and ER-negative status relates to epigenetic changes in breast cancer-related genes. To evaluate epigenetic differences in tumor-related genes relating to ER and HER2/neu status of primary tumors, we examined the promoter methylation status of the promoter region CpG islands of eight major breast tumor-related genes (RASSF1A, CCND2, GSPT1, TWIST, APC, NES1, RARβ2, and CDH1).

Methods

Paired ER-positive (n = 65) and ER-negative (n = 65) primary breast tumors (n = 130) matched for prognostic factors were assessed. DNA was extracted from paraffin-embedded tumor tissue after microdissection, and methylation-specific PCR and capillary-array electrophoresis analysis were performed.

Results

In early stages of tumor progression (T1 and N0), RASSF1A and CCND2 were significantly (P < 0.05) more methylated in ER-positive than in ER-negative tumors. GSTP1 hypermethylation was more frequent in the lymph node metastasis positive group than in the negative group. Double negative (ER-negative, HER2/neu-negative) breast cancers had significantly lesser frequencies of RASSF1A, GSTP1, and APC methylation (P < 0.0001, P < 0.0001, and P = 0.0035, respectively). Both ER and HER2/neu status correlated independently with these epigenetic alterations.

Conclusion

We demonstrated significant differences in tumor-related gene methylation patterns relevant to ER and HER2/neu status of breast tumors. This may be of significance in the assessment of targeted therapy resistance related to ER and HER2/neu status in breast cancer patients.     

Expression of Cell-Cycle Mediators in Ovarian Cancer Cells after Transfection with p16, p21, and p53

Objective.The purpose of this study was to determine whether transfection of ovarian cancer cell lines with recombinant adenoviral vectors containing wild-type p16^INK4a, p21^WAF1/Cip-1, and p53 caused growth inhibition and induction of apoptosis. We also measured the expression of the cell-cycle mediators Bax, Bcl-2, pRb, and mdm-2.Methods. We introduced the wild-type p16^INK4a, p21^WAF1/Cip-1, and p53 genes into the ovarian cancer cell lines SK-OV-3 (p16^INK4a and p53 null) and OVCA-420 (p16^INK4a and p53 wild-type) by adenoviral transfection. Cell growth inhibition was measured over a 10-day period. Induction of apoptosis was tested for both cell lines 48 h after cell transfection. Expression of cell-cycle mediators was evaluated by Western blot analysis and densitometry.Results. Growth inhibition was documented after transfection with p16^INK4a, p21^WAF1/Cip-1, and p53 in both SK-OV-3 cells and OVCA-420 cells. Apoptosis was greatest in SKOV-3 cells after transfection with p53. A significant expression of Bax was only seen in the SKOV-3 cells transfected with p53. The bcl-2 protein was poorly expressed in both cell lines. Expression of pRb was suppressed in OVCA-420 cells transfected with p16^INK4a and p21^WAF1/Cip-1. Infection with Adp16^INK4a and Adp53 led to an increase in the level of mdm-2 in the SK-OV-3 cell line only.Conclusions. In the ovarian cancer cell lines studied, cell growth was inhibited after transfection with p16^INK4a, p21^WAF1/Cip-1, and p53. Cell cycle arrest was highest with p53 transfection. The expression of pro-apoptosis proteins was primarily a function of p53 expression.     

Hyperfractionated split-course whole abdominal radiotherapy for ovarian carcinoma: tolerance and toxicity

Whole abdominal irradiation after chemotherapy and second look laparotomy for advanced ovarian carcinoma is poorly tolerated because of hematologic toxicity that frequently necessitates interruption or abandonment of treatment. A new treatment strategy using a hyperfractionated split course schedule to deliver a total of 30 Gy in 30 fractions over 6 weeks was designed in an attempt to overcome this problem, while not compromising the tolerance of late reacting normal tissues. Of 23 patients treated between August 1984 and June 1986, only one failed to complete therapy as scheduled. Six patients with gross residual disease also received a limited field boost of 15 Gy in 15 fractions after completion of treatment to the whole abdomen. None of these six patients achieved disease control, and five required surgery for intestinal obstruction with pathologic evidence of radiation bowel injury. Of the 17 patients who received no boost, five developed gut obstructions associated with tumor recurrence and not attributable to irradiation. We conclude that whole abdominal irradiation using the hyperfractionated split course schedule without a boost is safe and feasible but its therapeutic efficacy appears confined to subsets of patients with no visible residual disease at the time of second look laparotomy, or in whom all visible residual tumor can be resected.     

An epidemiological survey of recurrent abdominal pain in a rural Malay school

OBJECTIVE: To study the prevalence of complaints of recurrent abdominal pain (RAP) among school children aged 11-12 years in a rural setting in Malaysia. METHODOLOGY: Questionnaires were distributed to all parents and teachers of children aged 11-12 years who attended a small rural school in which all the children were Malays. Complaints of RAP were defined as at least three such complaints occurring over a period of at least 3 months. RESULTS: One hundred and sixty questionnaires were distributed, of which 148 were returned, giving a response rate of 92.5%. Sixty-one children (41.2%) had RAP. Approximately 45.2% of girls and 35.9% of boys reported having RAP. Compared with children without RAP, there was a significantly larger number of children with RAP (85.2%) who had at least one stress factor (P = 0.0109). There were no significant associations between RAP and total family income (P = 0.0573), a history of abdominal pain in at least one parent (P = 0.1686), a history of abdominal pain in at least one sibling (P = 0.0617), academic performance (P = 0.9967) or the degree of sports participation (P = 0.8469). There was an increased incidence of other systemic complaints in children with RAP when compared with children without RAP. CONCLUSION: Recurrent abdominal pain was found to be common among 11- to 12-year-old children in a rural Malay school. There was a significant association found between RAP and the presence of stressful events, as well as with the presence of other systemic complaints.     

Emergence and maintenance of actionable genetic drivers at medulloblastoma relapse

BackgroundLess than 5% of medulloblastoma (MB) patients survive following failure of contemporary radiation-based therapies. Understanding the molecular drivers of medulloblastoma relapse (rMB) will be essential to improve outcomes. Initial genome-wide investigations have suggested significant genetic divergence of the relapsed disease.MethodsWe undertook large-scale integrated characterization of the molecular features of rMB—molecular subgroup, novel subtypes, copy number variation (CNV), and driver gene mutation. 119 rMBs were assessed in comparison with their paired diagnostic samples (n = 107), alongside an independent reference cohort sampled at diagnosis (n = 282). rMB events were investigated for association with outcome post-relapse in clinically annotated patients (n = 54).ResultsSignificant genetic evolution occurred over disease-course; 40% of putative rMB drivers emerged at relapse and differed significantly between molecular subgroups. Non-infant MBSHH displayed significantly more chromosomal CNVs at relapse (TP53 mutation-associated). Relapsed MB_Group4 demonstrated the greatest genetic divergence, enriched for targetable (eg, CDK amplifications) and novel (eg, USH2A mutations) events. Importantly, many hallmark features of MB were stable over time; novel subtypes (>90% of tumors) and established genetic drivers (eg, SHH/WNT/P53 mutations; 60% of rMB events) were maintained from diagnosis. Critically, acquired and maintained rMB events converged on targetable pathways which were significantly enriched at relapse (eg, DNA damage signaling) and specific events (eg, 3p loss) predicted survival post-relapse.ConclusionsrMB is characterised by the emergence of novel events and pathways, in concert with selective maintenance of established genetic drivers. Together, these define the actionable genetic landscape of rMB and provide a basis for improved clinical management and development of stratified therapeutics, across disease-course.     

长期应用埃索美拉唑是维持已愈合糜烂性食管炎疗效及控制胃食管反流症状的有效方法

要确保糜烂性食管炎的治愈效果 ,长期抑制胃酸是一项必要措施。在不使用维持治疗时 ,已治愈的反流性或溃疡性食管炎 ,在 1年左右有近 80 %的病人会再出现食管黏膜的破损。 1年中奥美拉唑 2 0ｍｇ或兰索拉唑 30ｍｇ每天 1次治疗 ,可使复发率降至 30 %或更低。Ｈ2 受体阻滞剂 (Ｈ2 ＲＡ)维持治疗的效果不及ＰＰＩ。Ｈ2 ＲＡ 1年维持治疗的效果约为 50 %。奥美拉唑的Ｓ 异构体———埃索美拉唑是第一个发展为同分光学异构体的质子泵抑制剂 (ＰＰＩ)。埃索美拉唑 2 0或40ｍｇ剂量的血浆浓度时间曲线下面积 (ＡＵＣ值 )明显大于奥美拉唑 2 0ｍｇ …