目的 观察利妥昔单抗在治疗造血干细胞移植后血小板输注无效的有效性和安全性。方法 回顾分析我院2014年1月—2017年6月收治的11例利妥昔单抗治疗的造血干细胞移植后血小板输注无效的病例资料,其中包括重型地中海贫血8例,急性髓系白血病1例,重型再生障碍性贫血2例。结果 10例造血干细胞移植后血小板输注无效患者经利妥昔单抗治疗,375 mg/m²,每周1次,2~3次后血小板输注无效的状况明显改善;1例造血干细胞移植后血小板输注无效患者接受1次利妥昔单抗治疗,仍存在血小板输注无效,最终因颅内出血死亡。结论 利妥昔单抗是治疗造血干细胞移植后血小板输注无效的一种很有效的治疗方法。

Objective The purpose of our study was to evaluate the efficacy and safety of rituximab in the treatment of platelet transfusion refractoriness after hematopoietic stem cell transplantation. Methods We retrospectively analyzed 11paitents （8 thalassemia major,2 sever aplastic anemia,and 1 acute myeloid leukemia） with platelet transfusion refractoriness after hematopoietic stem cell transplantation. All 11 patients received treatment of rituximab. Results 10 of 11 platelet transfusion refractoriness patients after hematopoietic stem cell transplantation had improvement of platelets transfusion,1 patient of 11 platelet transfusion refractoriness patients had no response and died of intracranial hemorrhage. Conclusion Rituximab is a promising treatment in patients with platelet transfusion refractoriness after hematopoietic stem cell transplantation.

目的 在原来研究的基础上进一步研究Wnt-1信号通路蛋白-3（WISP-3）在高氧诱导肺上皮细胞凋亡中的作用。方法 通过Western blot检测和免疫组化检测不同肺上皮细胞中WISP-3的蛋白表达量。利用质粒转染和siRNA的方法在Beas-2B细胞中高表达和基因沉默WISP-3,通过细胞活性检测和流式细胞学技术检测高氧刺激后细胞的凋亡情况。结果 与空气对照相比,高氧刺激使肺上皮细胞的WISP-3蛋白表达量下降;WISP-3基因沉默或高表达使高氧诱导的肺上皮细胞凋亡增加或减少。结论 高氧刺激下,肺上皮细胞中WISP-3表达下降,WISP-3对高氧诱导的肺上皮细胞凋亡具有保护作用。

Objective To explore how Wnt-1 inducible signaling pathway protein-3 （WISP-3） participate in and play a regulatory role in the process of hyperoxia induced apoptosis in lung epithelial cells. Methods The expression of WISP-3 was detected via Western blot and immunohistochemistry. High expression and low expression of WISP-3 were performed by plasmid transfection and siRNA. Cell viability and flow cytometry were executed to detect the hyperoxia-induced apoptosis in Beas-2B. Results Compared to the group of air control,the expression of WISP-3 protein in lung epithelial cells decreased obviously after hyperoxia. Cell survival decrease and apoptosis increased after hyperoxia in Beas-2B cells with low expression of WISP-3. Vice versa. Conclusion The expression of WISP-3 decreased after hyperoxia in lung epithelial cells. The role of WISP-3 in this process may be protective.

目的 探讨妊娠期高血糖患者母体胰岛β细胞自身抗体对新生儿的影响。方法 选取2014年3月—2016年3月于我院就诊的口服75 g葡萄糖耐量试验(OGTT)异常的孕妇共276例,以60例健康孕妇作为对照组,分别在产前和OGTT试验后空腹抽取静脉血进行检测,检测指标包括谷氨酸脱羧酶抗体(GADA)、胰岛细胞抗体(ICA) )和胰岛素自身抗体(IAA),记录每个出生患儿的体重、Apgar评分、血糖指标进行分析。结果 276例GDM患者中34.41%至少存在一种相关抗体阳性,GDM组GADA阳性、IAA阳性和阴性的搏动指数异常比例均高于健康组(P<0.05),IAA阳性的胎儿生长受限比例比例较GDM组抗体阴性的比例高(P<0.05),妊娠晚期胰岛细胞抗体阳性(OR:6.41,95%CI:1.41~34.13)均为新生儿产后进入ICU进行监护的风险因素,妊娠中、晚期GADA阳性(OR:10.26,95%CI:1.42~75.14) 和妊娠晚期GADA阳性(OR:8.15,95%CI:1.43~46.83)均为新生儿窒息的风险因素 。结论 妊娠晚期胰岛细胞抗体阳性为新生儿产后进入ICU进行监护的风险因素,妊娠中、晚期GADA阳性是新生儿窒息的风险因素。

目的 探讨精液白细胞和形态正常精子百分率的相关性。方法 随机选取2014年4月—2015年8月来生殖中心进行就诊的男性800例,分析患者精液中的白细胞数量以及分析患者的精子形态,比较二者的相关性。结果 正常形态精子百分率低于正常参考值的患者中有61.9％的白细胞数大于1×10⁶,精子形态正常的患者有38.9％患者的白细胞数大于1×10⁶,患者比例差异有统计学意义(P<0.05)。精子形态正常的患者(320例)和正常形态精子百分率低于正常参考值的患者(480例)相比,精子形态正常的比例远远大于正常形态精子百分率低于正常参考值的患者(P<0.05)。而正常形态精子百分率低于正常参考值的患者的头部异常、颈部和中间部分异常、尾部异常精子比例高于正常组(P<0.05)。正常形态精子百分率低于正常参考值的和正常形态的精子比例呈正相关;和大头精子百分率、锥形以及小头形态精子百分率呈负相关;和梨形以及其他形态的精子呈正相关。结论 精液中的白细胞可以影响精子的参数,使得患者精液中的颈部或中部异常形态、头部和尾部异常形态的精子比例提升,具体机制有待进一步研究。

目的 探讨Reversine对人肝星状细胞系LX-2凋亡的影响。方法 设对照组和Reversine干预组,其中Reversine干预组分为7个浓度,分别为1,5,10,20,40,80,120 μg/mL,CCK-8法检测Reversine对LX-2增殖的影响,选取最佳浓度。将细胞重悬在加入5 μL FITC-Annexin V和5 μL PI,用流式细胞仪进行了凋亡率分析,免疫荧光检测凋亡蛋白bcl-2及caspase 3。结果 Reversine可促进LX-2细胞凋亡,随着Reversine浓度增加,LX-2的凋亡可呈剂量依赖关系,其中10 μg/mL为最佳浓度,LX-2细胞的bcl-2蛋白的表达显著下降而cleaved-caspase 3的表达显著上升。结论 Reversine可通过促进caspase-3蛋白活化、抑制bcl-2蛋白表达的方式诱导LX-2凋亡。

目的 评价鞘内注射雷帕霉素对CCI神经病理性痛大鼠的痛阈及脊髓背角胶质细胞表达的影响。方法 健康雄性SD大鼠30只随机分为6组:①CCI组:CCI术后14天处死;②正常对照组:不做任何处理; ③前对照剂组:鞘内置管3天后行CCI术,术后4小时后鞘内给同体积生理盐水,连给3天; ④前给药组:鞘内置管3天后行CCI术,术后4小时鞘内给雷帕霉素溶液,连给3天; ⑤后对照剂组:鞘内置管3天后行CCI术,术后7天鞘内给同体积生理盐水,连给3天;⑥后给药组:鞘内置管3天后行CCI术,术后7天鞘内给雷帕霉素溶液,连给3天。各组于CCI术前1天和术后第2、4、6、8、10、12、14天测机械痛阈和热痛阈。术后14天测痛后用多聚甲醛灌注大鼠,取L4~5脊髓,免疫组化染色,星形胶质细胞标记蛋白(GFAP)检测星形胶质细胞表达变化,并定量分析。结果 与对照组相比,CCI手术组热痛阈和机械痛阈从CCI手术后第4天开始下降(P<0.05);前后给药对照剂组与CCI组相比,差别无统计学意义(P>0.05)。前给药组痛阈从CCI手术后第4天开始上升并持续至手术后第14天,与CCI组相比,差别有统计学意义 (P<0.05)。与CCI组相比,后给药组痛阈从CCI第8天开始上升并持续至手术后第14天,差别有统计学意义(P<0.05)。 与正常对照组比较,CCI组、前、后对照剂组手术侧脊髓背角GFAP染色阳性区平均光密度与阳性面积均有增加,差别有统计学意义(P<0.05)。前、后给药组手术侧GFAP染色阳性区平均光密度与阳性面积与CCI组比较,均有明显降低,差别有统计学意义(P<0.05)。结论 鞘内注射雷帕霉素可缓解大鼠神经病理性痛,并抑制脊髓背角胶质细胞的激活。

Objective To evaluate the effects of intrathecal injection of rapamycin on pain threshold and spinal cord gliacyte activation in rats of neuropathic pain. Methods Healthy 30 male SD rats were randomly divided into 6 groups(n=5 in each group): ① control group without operation or intrathecal injection. ②CCI group without intrathecal injection. ③ intrathecal injection of rapamycin 10 μg(10 μL) 4 hours after CCI operation and the next 2 days once a day. ④ intrathecal injection of NS10 μL 4 hours after CCI operation and the next 2 days once a day. ⑤ intrathecal injection of rapamycin 10 μg(10 μL) 7 days after CCI operation and the next 2 days once a day.⑥ intrathecal injection of NS10 μL 7 days after CCI operation and the next 2 days once a day. Mechanical and thermal threshold were tested 1 day before the CCI operation and 2^th、4^th、6^th、8^th、10^th、12^th、14^th days after the CCI operation for all the rats. Lumbar segment of spinal cords was removed for determination of glial fibrillary acidic protein(GEAP) in spinal cord by immuohistochemistry dyeing and assay in the 14th day after CCI operation for all the rats. Results Mechanical and thermal hyperalgesia emerged on 4th day and maintained till 14th day after CCI operation(P<0.05). After intrathecal injection of rapamycin 4 hours or 7days after CCI, mechanical and thermal threshold significantly increased compared to intrathecal injection of NS(P<0.05). And the sum area of GFAP positive and the mean density of GFAP positive area in the dorsal horn of operation side greatly increased in rapamycin treated groups compared NS treated groups(P<0.05). Conclusion Intrathecal injection of rapamycin may attenuate CCI induced hyperalgesia and inhibit the activation of astrocyte.

目的 探索蟾毒灵对舌鳞状细胞癌Tca8113细胞增殖、凋亡的影响及可能作用机制。方法 以人舌鳞癌Tca8113细胞为研究对象,MTT法检测10、20、40、80、160 nmol/L浓度蟾毒灵体外抑制Tca8113细胞增殖的活性;检测蟾毒灵干预下肿瘤细胞Na⁺-K⁺-ATP酶活性的变化;Western blot发检测Bcl-2、Bax、caspase-3蛋白表达。结果 蟾毒灵有抑制Tca8113细胞的活性,且呈剂量-时间依赖性;在蟾毒灵干预下Tca8113细胞Na⁺-K⁺-ATP酶收到抑制;Western blot结果显示凋亡相关Bax、caspase-3蛋白表达上调,Bcl-2蛋白表达下调。结论 蟾毒灵通过抑制细胞膜Na⁺-K⁺-ATP酶活性,通过调节Bcl-2凋亡通路的相关蛋白,最终激活caspase-3,诱导人舌鳞癌Tca8113细胞凋亡。

Objective To explore the effects of bufalin on proliferation and apoptosis of tongue squamous cell carcinoma Tca8113 cells and its possible mechanism. Methods Tca8113 cells were treated with 10, 20, 40, 80 and 160 nmol/L Tca8113 cells in vitro. MTT assay was used to detect the inhibitory effect of bufalin on the proliferation of Tca8113 cells; And the activity of Na⁺-K⁺-ATPase in tumor cells was detected by the interference of bufalin; The expression of Bcl-2, Bax and caspase-3 protein was detected by Western blot. Results Bufalin inhibited the activity of Tca8113 cells in a dose-and time-dependent manner; Na⁺-K⁺-ATPase in Tca8113 cells was inhibited by bufalin; The results of Western blot showed that the expression of Bax and caspase-3 protein was up-regulated and the expression of Bcl-2 protein was down-regulated. Conclusion Bufalin induced the apoptosis of human tongue squamous cell carcinoma Tca8113 cells by inhibiting the activity of Na⁺-K⁺-ATPase and regulating the related proteins of Bcl-2 apoptosis pathway, finally activating caspase-3.

目的 对比分析巨幼细胞性贫血和难治性贫血患者骨髓形态的异同及其临床意义。方法 选取我院2010年4月—2016年4月收治的巨幼细胞性贫血(MA)35例,难治性贫血(RA)35例为研究对象。收集两组患者临床检验及检查资料,对比分析其临床表现、骨髓检查结果及血液检查等结果。结果 MA组患者红系病态率、粒系病态率、淋巴样小巨核率及PAS阳性率均明显低于RA组,差异明显(P<0.05);两组患者在发热、出血、消化系反应等临床表现率上相比差异不明显(P>0.05);两组患者血象检查中血红蛋白、血小板、及白细胞含量相似,两组差异不明显(P＞0.05)。结论 巨幼细胞性贫血与难治性贫血临床表现及外周血象相似,RA骨髓形态应重点观察淋巴样小巨核细胞;MA组则应强调红系巨幼变的胞体。

目的 研究雷公藤红素对人阿霉素耐药MCF-7/ADR乳腺癌细胞生长的作用。方法 采用MTT试验检测MCF-7/ADR细胞对阿霉素的耐药情况以及雷公藤红素对MCF-7/ADR细胞生长的影响;采用Annexin V-FITC/PI双染试验分析雷公藤红素对MCF-7/ADR耐药细胞凋亡的诱导作用;应用流式细胞周期分析检测雷公藤红素对MCF-7/ADR耐药细胞周期的影响。结果 MCF-7/ADR细胞对阿霉素耐药指数达14.54;而雷公藤红素能有效抑制阿霉素耐药细胞MCF-7/ADR的生长,并呈现浓度依赖性,作用48 h的IC50是1.04 μmol/L,MCF-7/ADR对雷公藤红素的耐药指数仅为0.87。2 μmol/L雷公藤红素作用8 h后,Annexin V-FITC染色阳性的MCF-7/ADR细胞比例较对照显著升高,差异有统计学意义(P＜0.05)。在1 μmol/L雷公藤红素作用24 h后,G1期细胞比例由对照(59.22±3.78)%升高至(77.44±4.21)%,而S期细胞比例由对照(37.51±2.91)%降至(19.65±2.25)%,差异有统计学意义(P＜0.05)。结论 雷公藤红素能激活MCF-7/ADR细胞凋亡的发生,并诱导MCF-7/ADR发生 G1/S细胞周期阻滞,从而对阿霉素耐药MCF-7/ADR细胞的生长发挥高效抑制作用。

Objective To investigate the effect of celastrol on the growth of adriamycin-resistant MCF-7/ADR breast cancer cells. Methods The resistance situation of MCF-7/ADR cells to adriamycin and the effect of celastrol on the growth of adriamycin-resistant MCF-7/ADR cells were evaluated by MTT assay. The effect of celastrol on apoptosis in MCF-7/ADR breast cancer cells was determined by annexin V-FITC/PI double staining. The effect of celastrol on cell cycle progression was determined by flow cytometry analysis. Results The resistance index of MCF-7/ADR cells to adriamycin was 14.54. After treatment with celastrol for 48 h, MCF-7/ADR cells displayed markedly inhibited growth in a dose-dependent manner, and calculated IC50 was 1.04 μmol/L. Celastrol decreased the resistance index of MCF-7/ADR from 14.54 to 0.87. The numbers of apoptotic MCF-7/ADR cells, as revealed by annexin V binding, significantly increased upon celastrol treatment (P＜0.05). Celastrol treatment caused an increase of cells in G1 phase from (59.22±3.78)% to (77.44±4.21)%, while the percentage of cells in S phase was decreased from(37.51±2.91)% to(19.65±2.25)%(P＜0.05). Conclusion These data demonstrated that celastrol induced apoptosis and G1/S cell cycle arrest in MCF-7/ADR cells and consequently displayed potent cytocidal effect on adriamycin-resistant MCF-7/ADR breast cancer cells.

目的 观察紫河车提取物联合顺铂对人脑胶质瘤细胞增殖与凋亡的影响。方法 把正常培养传代后的U251胶质瘤细胞按随机分配的方法分为四组,A组仅加普通培养液,B、C、D组各加紫河车提取物(400 mg/mL)2 mL、顺铂(1 mg/mL)0.01 mL、紫河车提取物(400 mg/mL)2 mL＋顺铂(1 mg/mL)0.01 mL;MTT法观察U251细胞增殖情况,流式细胞仪检测U251细胞凋亡率。结果 培养12、24、36、48、60 h,B、C、D组细胞增殖指数逐渐下降,与A组进行比较,各组P值均小于0.05;其中,将D组与B、C组进行比较,P值小于0.05。将各组培养24 h后上机,测得A、B、C、D各组细胞的凋亡率分别为(0.3±0.2)%,(10.6±1.5)%,(35.9±2.8)%,(52.1±4.1)%。其中,B、C、D各组和A组进行比较,P值均小于0.05;将D组与B、C组两组进行比较,P值也均小于0.05。结论 紫河车提取物联合顺铂可抑制人脑胶质瘤U251细胞增殖,并诱导其凋亡。

Objective To observe the effect of cisplatin combinated with the placental immunoregulating polypeptide (PIP) on proliferation and apoptosis of glioma cells. Methods Randomly we divide the normal handed U251 glioma cells into four groups. We added ordinary nutrient solution to group A, while added activated PIP(400 mg/mL)2 mL to group B, cisplatin (1 mg/ml) 0.01 ml to group C, PIP 400 mg/mL)2 mL and cisplatin (1 mg/mL) 0.01 mL to group D. We surveyed the proliferation rate of gliobma cells by MTT experimental method and analyzed the apoptosis of U251 glioma cells by flow cytometry. Results The index of cell proliferation of group B,C,D declined gradually with the training of 12 h,24 h,36 h,48 h,60 h. Compared B, C,D group with A group, P<0.05,and compared group D with group B and group C, P< 0.05. Put groups culturing of 24 hour on flow cytometer, the glioma cells apoptosis rate of each group was 0.3%±0.2%、10.6%±1.5%、35.9%±2.8%、52.1%±4.1% respectively. Compared group B,C,D with group A, P<0.05,and compared group D with group B and group C, P<0.05. Conclusion Placental immunoregulatingpPolypeptide combined with cisplatin may restrain the proliferation of human glioma cells, meanwhile increase the apoptosis of glioma cells.