目的 探讨单倍体亲缘异基因造血干细胞移植治疗重型再生障碍性贫血(SAA)的可行性。方法 对1例诊断SAA 4年余,先后经CsA治疗、脐血移植治疗均无效并反复输注红细胞、血小板的12岁男性患者进行单倍体亲缘异基因造血干细胞移植,供者为其胞兄,高分辨HLA基因型5/10相合,预处理方案为BU+CTX+ATG:BU 3.2 mg/kg×2 d,CTX 50 mg/kg×4 d,ATG 2.5 mg/kg×4 d。干细胞来源为G-CSF动员的骨髓+外周造血干细胞,共计输注单个核细胞(MNC)4.055×10⁸/kg(受者体重),CD 34 2.331×10⁶/kg。GVHD预防:-1 d采用与受者HLA部分相合的第三方脐带血细胞,术后联合应用环孢素A、短程氨甲碟呤、霉酚酸酯。结果 造血缓慢重建,术后22天(+22 d)ANC>0.5×10⁹/L,术后3月血小板脱离输注。+26天DNA指纹图全部表现为供者基因型。+40天血型转为供者型“O”型。+29 d出现急性移植物抗宿主病aGVHD(胃肠型,Ⅲ度),+31 d、+34 d及+42 d予巴利昔单抗20 mg静滴,+40 d、+44 d、+63 d输注间充质干细胞,患者急性GVHD逐渐控制。期间曾出现肺部感染、口腔黏膜炎及巨细胞病毒血症,经抗感染后可控制。现随访3年,血象正常稳定,Kamofsky评分100分。结论 单倍体亲缘异基因造血干细胞移植治疗SAA,对无相合供者(包括亲缘或非亲缘)且强效免疫抑制治疗失败的患者,可考虑进行,GVHD和感染为主要并发症,需根据患者病情采用相应措施。

Objective To investigate the feasibility of haploid genetic allogeneic hematopoietic stem cell transplantation in the treatment of severe aplastic anemia(SAA) in our hospital. Methods A 12-year-old patient with acquired SAA for 4 years showed no response to CsA and cord blood transplant treatment and was transfusion-dependent. Lacking an HLA-identical sibling donor, the patient was treated with HSCT from his brother 5/10 matched at the generic level. Theconditioning regimen was BU+CTX+ATG:BU 3.2 mg/kg×2 d,CTX 50 mg/kg×4 d,ATG 2.5 mg/kg×4 d. Stem cells were the source of G-CSF mobilization of bone marrow and peripheral blood stem cells, dose of stem cells infused: mononuclear cells (MNC) 4.055×10⁸/kg (body weight of subject), CD34 2.331×10⁶/kg. Prevention of GVHD: -1 d Third-party umbilical cord blood cells which were HLA partially matched were used. Postoperative joint use included cyclosporine A, short-course methotrexate, mycophenolate mofetil. Results Hematopoiesis was slowly rebuilding, 22 d after surgery (+22 d) ANC> 0.5×10⁹/L, after three months departing from transfusion of platelets. +26 d suggesting that the DNA fingerprints showed donor genotypes. +40 d into donor blood type “O” type. + 29 d occurred acute GVHD (GI type, Ⅲ degrees), + 31 d, + 34 d + 42 d infusion of basiliximab 20mg, + 40 d, + 44 d, + 63 d infusion of mesenchymal stem cells. Gradually acute GVHD was controlled in the patient, who had lung infections, oral mucositis and cytomegalovirus viremia, could be controlled with anti-infective. Now followed up for 3 years, hemogram change has been normal and stable. Kamofsky score was 100 points. Conclusion It may be considered to have haploid genetic allogeneic hematopoietic stem cell transplantation for treatment of SAA, for those patients who have non-matched donor (including relatives and non-relatives) and potent immunosuppressive therapy failure. GVHD and infection are major complications. Need to adopt appropriate measures in accordance with the patient's condition.

目的 探讨同步放化疗治疗晚期非小细胞肺癌(NSCLC)的临床疗效。方法 选取我院2014年收治的晚期NSCLC患者102例,随机分为观察组和对照组,对照组以紫杉醇联合顺铂化疗方案(TP)化疗治疗,观察组加用三维适形放疗同步放化疗治疗,观察两组的临床疗效和不良反应。结果 观察组和对照组患者的总有效率分别为43.14%和21.57% (P<0.05),两组患者的总控制率分别为84.31%和66.67% (P<0.05)。两组患者Ⅲ～Ⅳ级胃肠道反应、白细胞减少、骨髓抑制的发生率差异无统计学意义(P>0.05),观察组比对照组增加了放射性肺炎和放射性食管炎的发生 (P<0.05)。两组患者生活质量比较,差异具有统计学意义(P＜0.05)。结论 同步放化疗治疗晚期NSCLC可以显著提高治疗的总有效率、肿瘤的控制率及生活质量,但也使不良反应增加,选择治疗方案时应根据个体情况综合考虑。

Objective To study the clinical efficacy of concurrent chemoradiotherapy in the treatment of advanced non small cell lung cancer (NSCLC). Methods 102 cases of advanced NSCLC treated in our hospital in 2014 were selected and randomly divided into the observation group and the control group. Patients in control group were treated by chemotherapy with Paclitaxel combined Cisplatin (TP), while those in observation group were treated by concurrent chemoradiotherapy with three- dimensional conformal radiotherapy and TP. The clinical efficacy and adverse reactions of the two groups were observed. Results The total effective rate of the observation group and the control group were 43.14% and 21.57% (P<0.05), respectively. The total control rate of the two groups were 84.31% and 66.67%(P<0.05). In two groups III ~ IV gastrointestinal tract reaction, leukopenia, bone marrow suppression occurrence had no statistical significance(P>0.05). Incidence of radiation pneumonitis and radiation esophagitis increased in observation group(P<0.05). The difference of quality of life between the two groups was statistically significant(P<0.05). Conclusion Concurrent radiotherapy and chemotherapy in the treatment ofadvanced non-small cell lung cancer(NSCLC)can not only significantly improve the treatment, tumor control rate and quality of life, but also make adverse reaction increased. Treatment options should be chosen based on individual circumstances into account

目的 探讨自噬激活剂和自噬抑制剂分别对鼻咽癌细胞CNE2放疗敏感性的影响。方法 利用RNA干扰技术使atg5基因沉默,构建自噬抑制细胞模型后,与雷帕霉素、氯喹分别处理的两组细胞一起,每天以X射线5Gy照射细胞,连续8天观察各组细胞的生长状况,并设置对照组。以MTT法及克隆集落形成法检测其细胞活力,用流式细胞仪分析其细胞周期。结果 与对照组相比,其他三组细胞存活率、克隆形成率、照射后存活率均显著降低(P<0.05);细胞周期检测除对照组外其他三组细胞集中在G0/G1期,其他两个时期比G0/G1期相对较少。结论 自噬抑制剂与激活剂和atg5沉默均能为CNE2放疗增敏,然而自噬激活剂的增敏效果好于其他,为增敏放疗提供实验依据,开辟新的放疗增敏途径。

Objective This study aimed to investigate the autophagy activators and inhibitors effects in nasopharyngeal CNE2 cells radiotherapy sensitization. Methods Atg5 gene silencing by RNA interference technology, two groups of cell autophagy inhibition were built by rapamycin and chloroquine respectively. Then 5Gy x-ray irradiation of cells was taken every day, after 8 days in a row in each group of cell growth and setting a control group. The cell viability was clonaled colony formation by MTT method assay and cell cycle by flow cytometry analysis. Results The three cell group survival rate, colony-forming rate and survival after irradiation were significantly lower (P<0.05) than the control group. Detection of cell cycle in addition to control three other groups concentrated in the G0/G1 period.That of two other periods was relatively fewer than that of the G0/G1 period. Conclusion Autophagy inhibitors, activators and atg5 silence improved the radio-sensitization to CNE2. The autophagy activator group improving the sensitivity was better than the others.This study provided evidence to sensitive radiotherapy, explored a new promising radiosensitization ways.

目的 探讨初诊的弥漫大B细胞淋巴瘤(DLBCL)患者外周血Th17细胞的表达与国际预后指标(IPI)之间关系。方法 初诊DLBCL组(n=45)按照国际预后指数(IPI)积分分为4组,采用ELISA和流式细胞术检测各个DLBCL组与正常对照组(n=43)的外周血中IL-17的浓度以及Th17阳性细胞比例,比较各组数值间的差异,并分析IPI的5个指标与IL-17的浓度以及Th17阳性细胞比例的相关性。结果 DLBCL中高危组与高危组的IL-17的浓度以及Th17阳性细胞比例较正常对照组及其他IPI组降低,有显著性差异;DLBCL四组的IL-17的浓度以及Th17阳性细胞比例均低于正常对照组;且可见随着IPI分组的增高,IL-17的浓度以及Th17阳性细胞比例呈降低的趋势;IPI指标中年龄、临床分期、全身状态与Th17细胞的表达有相关性。结论 初诊DLBCL患者外周血Th17细胞的表达与国际预后指标有关系;随着IPI积分的增加,DLBCL患者Th17细胞表达下降;临床上对于年龄60岁以上、临床分期Ⅲ期以上、长期卧床及需别人照顾的患者更要注意监测其外周血Th17细胞的表达情况。

Objective To explore the relationship between the international prognosis indexes(IPI) and the Th17 cells expression in DLBCL patients. Methods DLBCL patients (n=45) were divided into 4 groups according to IPI score, peripheral blood were taken from each person in DLBCL groups and normal group. We used ELISA to test IL-17 and flow cytometry (FCM) to examine the Th17 positive cells. We compared the value of each group, and analyzed the relativity of IPI and Th17 cells' expression. Results Th17 cells' expression level in middle-high risk group and high risk group were higher than that in normal group and other IPI groups; Th17 cells' expression level in DLBCL groups were all lower than that in normal group; Th17 cells decreased while IPI score increased; Age, clinical stage and general body state have the relativity with DLBCL patients' Th17 cells expression level. Conclusion Th17 cells in DLBCL patients has the relativity with prognosis index. In our clinical diagnosis and treatment, we need to pay more attention to those patients who are over 60 years old, or whose clinical stage is above Ⅲ phase, or who need to stay on bed for a long time and need other peoples' help .

目的 构建重组pEGFP-C3-HCVc真核表达载体,并建立稳定表达HCVc基因的肝内胆管癌细胞株RBE-core。方法 采用PCR钓取目的基因HCVc,并克隆入pEGFP-C3的多克隆位点,构建pEGFP-C3-HCVc重组质粒。经过双酶切及测序验证后,采用脂质体将pEGFP-C3-HCVc质粒转染到RBE细胞中,经2周G418 (200 μg/mL) 筛选后进行单克隆挑选及扩大培养,建立稳定表达HCVc的胆管癌细胞株RBE-core。采用RT-PCR和Western blot验证HCVc在RBE-core中的表达情况。结果 PCR成功钓取HCVc基因,大小约573 bp,并插入pEGFP-C3载体HindⅢ和BamHⅠ多克隆位点;双酶切及测序证实目的基因HCVc正确连接到pEGFP-C3的多克隆位点。RT-PCR和Western blot分别在573 bp处和34 KD左右检测到相应的阳性条带。结论 成功构建重组质粒pEGFP-C3-HCVc,并在胆管癌细胞RBE中获得稳定表达。

Objective To construct a recombinant plasmid of pEGFP-C3-HCVc containing hepatitis C virus core protein, and establish the HCVc-expressing cell line RBE-core. Methods The HCVc gene was amplified by PCR and cloned into HindⅢ and BamHⅠsite of pEGFP-C3 plasmid. The recombinant plasmid of pEGFP-C3-HCVc was confirmed by sequencing. RBE cells were transfected with the recombinant plasmid by using Lipofectamine 2000, and then performed G418 (200 μg/mL) selection after 2 weeks. The expressing of HCVc gene in RBE cells was confirmed by RT-RCR and western blot. Results The recombinant plasmid of pEGFP-C3-HCVc was successfully constructed. RT-PCR and western blot detected a 573bp and 34KD bland, indicating the stably expressing of HCVc in RBE cells. Conclusion The recombinant plasmid of pEGFP-C3-HCVc is stabled expressing in RBE cells,which provides support for the further study.

目的 研究灵芝孢子油抑制肿瘤细胞增生的作用。方法 通过“预防性”和“治疗性”喂食方法,观察并检测喂食过程中小鼠乳腺癌细胞生长速度及处死后的瘤重量。结果 灵芝孢子油喂食组肿瘤生长速度较对照组慢,且预防性喂食组抑瘤效果好于“治疗性”喂食组。结论 灵芝孢子油具有抑制小鼠乳腺癌生长的作用。

Objective To research the effect of tumor growth inhibition by Ganoderma spore oil. Methods Murine mammary cancer cells were inoculated and Ganoderma spore oil was given by preventing and therapeutic feeding respectively. Results Tumor growth speed of Ganoderma spore oil feeding group was slower than control group, and tumor weight was lighter than control group. In addition, tumor weight of preventing feeding was lighter than therapeutic feeding. Conclusion Ganoderma spore oil has the effect of anti - mammary cancer cell growth.

目的 研究前列腺癌细胞中miR-221的表达情况及其对癌细胞增殖的影响。方法 运用实时荧光定量PCR(qRT-PCR)检测miR-221在前列腺正常细胞株与前列腺癌细胞株中表达的差异情况,利用细胞转染构建miR-221过表达LNCaP和DU145细胞株,再通过CCK8细胞增殖实验检测细胞增殖情况的变化。结果 qRT-PCR检测细胞株发现miR-221在PC3、LNCaP和DU145三种前列腺癌细胞株中表达量均比前列腺正常细胞株PrEC低 (F=254.197,P<0.001),其中两两比较差异也均有统计学意义。细胞转染技术构建的miR-221过表达LNCaP和DU145细胞株,经qRT-PCR结果显示,miR-221在LNCaP和DU145细胞株中的表达水平明显升高(LNCaP,倍数变化=2.24,t=3.46,P<0.01;Du145,倍数变化=2.24,t=4.29,P<0.01)。细胞增殖实验结果显示,过表达了miR-221的LNCaP(P<0.001)和DU145(P<0.001)细胞生长速度慢于对照组。结论 实验证明miR-221表达过度能减慢前列腺癌细胞的增殖,miR-221有可能成为前列腺肿瘤治疗的生物学标志物。

Objective To investigate miR-221 expression in prostate cancer cells and its influence on prostate cancer cell proliferation. Methods miR-221 expressions in prostate normal cell lines and cancer cell lines were measured by qRT-PCR. Overexpression of the miR-221 in LNCaP and DU145 cell lines used by cell transfection. Effects of the depletion on cell proliferation were assessed in vitro with CCK8. Results qRT-PCR showed miR-221 was lower expressed in PC3, LNCaP and DU145 than in PrEC(F=254.197, P<0.001), in which pairwise comparison also had significant differences. qRT-PCR showed miR-221 expression rose significantly in LNCaP and DU145 cell lines whose miR-221 was overexpression with cell transfection(LNCaP, Fold Change=2.24,t=3.46,P<0.001;Du145, Fold Change=2.24,t=4.29,P<0.001). Cell proliferation assay showed that growth of LNCaP(P<0.001) and DU145(P<0.001) cells whose miR-221 was overexpression was slower than the control group. Conclusion This study demonstrates miR-221 overexpression can inhibited the proliferation of prostate cancer cells for the first time, it also suggests that miR-221 has the potential to serve as a biomarker for PCa therapy.

目的 氧化苦参碱对视网膜母细胞瘤细胞SM-106凋亡的诱导作用及机制。方法 以不同作用时间(24 h、48 h、72 h)和不同作用浓度(12.5 μl/mL、25 μl/mL、50 μl/mL、100 μl/mL)氧化苦参碱处理视网膜母细胞瘤细胞SM-106,分别采用流式细胞仪及western blot检测视网膜母细胞瘤细胞SM-106细胞凋亡及其凋亡因子(Bax、Bcl-2)蛋白表达。结果 氧化苦参碱可促进SM-106细胞体外凋亡,上调Bax蛋白表达及Bax/Bcl-2蛋白表达比值,下调Bcl-2蛋白表达,并呈现剂量及时间依赖性。结论 氧化苦参碱可诱导视网膜母细胞瘤细胞SM-106凋亡,调控凋亡因子Bax、Bcl-2的表达是其可能作用机制。

Objective To evaluate the apoptosis and its mechanism of retinoblastoma cells SM-106 induced by oxymatrine. Methods Retinoblastoma cells SM-106 were treated with different time(24 h、48 h、72 h)and different concentrations(12.5 μl/mL, 25 μl/mL, 50 μl/mL or 100 μl/mL) of oxymatrine. The apoptosis and protein expression of apoptosis factors (Bax and Bcl-2) were respectively determined by flow cytometry and western blot. Results Oxymatrine significantly promoted the SM-106 cells apoptosis in vitro, raised Bax protein expression and Bax/Bcl-2 protein expression ratio, reduced the Bcl-2 protein expression, and showed the dose and time dependent. Conclusion Oxymatrine is able to induce the apoptosis in retinoblastoma cells SM-106. Regulating apoptosis related gene Bax and Bcl-2 expression may be the mechanism of apoptosis.

目的 调查I-IIa期宫颈鳞状细胞癌患者术后随诊和复发情况,并探索随诊和复发的相关因素。方法 分析2011年1月—2012年6月中山市妇幼保健院收治的128例宫颈鳞状细胞癌患者的临床资料,分析他们的术后随诊情况及其影响因素。将有液基薄层细胞学(TCT)及盆腔检查者视为有效随诊的患者,分析肿瘤复发情况及其影响因素。结果 128例患者中,有效随诊104例(占总人数的81.25%)。年龄组越小,随诊率越高(P=0.034);城市患者较农村患者,随诊率高(P=0.028);文化程度越高,随诊率越高(P=0.028);职业为公务员、教师、公司职员或自由职业、个体,随诊率比职业为家庭妇女和工人更高(P=0.030)。在104例随诊者中,2年复发8例(占7.7%)。临床分期较晚,肿瘤直径>4cm,病理类型,脉管内瘤栓阳性,淋巴结转移阳性是影响患者2年复发的危险因素(P<0.05)。结论 临床分期,肿瘤直径,脉管内瘤栓阳性,淋巴结转移等临床病理特征与患者复发密切相关。应针对宫颈癌患者社会人口学特点制定措施,提高随诊率。

Objective To investigate the follow-up rate after surgical treatment and the recurrence rate among patients with cervical squamous cell carcinoma in I-IIa stage, and identify associated factors. Methods The clinical data of 128 cases of cervical squamous cell carcinoma from January 2011 to June 2012 in Zhongshan maternal and child health hospital was collected. Follow up record of these cases was analyzed and factors associated with follow up were identified. Patients having taken thinprep liquid-based cytology test (TCT) and pelvioscopy were treated as cases being followed effectively, and their recurrence and associated factors were analyzed. Results Among the 128 patients, 104(81.25%) were effectively followed. Patients with younger age(P=0.034), living in city (P=0.028), with higher education level(P=0.028) had higher follow-up rate. Civil servants, teachers and freelancers had higher follow-up rate than housewives and workers (P=0.030).Eight of the 104 effectively followed cases recurred within two years after surgery, accounting for 7.7%. The associated factors of recurrence included clinical stage, tumor diameter, larger than 4cm, the depth of myometrial invasion, deeper than 2/3, vascular tumor thrombus positive, and lymph node metastasis positive (P<0.05). Conclusion Clinical stage, tumor size, vascular tumor thrombus positive, lymph node metastasis positive were associated with recurrence. Tailored strategies should be developed to improve the follow up rate based on the social and demographic characteristics of patients.