HLA-DPB1 gene polymorphism and multiple sclerosis: a large case-control study in the southwest of France

The polymorphism at the HLA-DPB1 locus has been characterized in a large number of patients with multiple sclerosis (n = 112) and in healthy controls (n = 115). Both patients and controls lived in the southwest of France (in the Pyrénées Atlantiques) and had similar ethnic background. The typing procedure involved the selective amplification of the second exon of the DPB1 locus by polymerase chain reaction, followed by hybridization of the amplified DNA with 14 sequence-specific oligonucleotide probes. Individual alleles were identified by the pattern of hybridization of the different probes. The distribution of the DPB1 alleles was not significantly different in multiple sclerosis patients and controls (p = 0.11). This does not corroborate the reported association of multiple sclerosis with the primed lymphocyte typing (PLT)-defined DPw4 specificity and is not in favour of a role played by polymorphic residues of the DP molecule in susceptibility to multiple sclerosis.     

卵巢未成熟畸胎瘤43例综合治疗疗效分析

为探讨不同手术方式及化疗方法对卵巢未成熟畸胎瘤的疗效及相关影响因素，对43例卵巢未成熟畸胎瘤患者进行回顾性分析。手术方式分为：保守性手术（患侧附件、大网膜切除术＋盆、腹腔腹膜结节切除术＋盆腔淋巴结清扫术）及根治性手术（全子宫、双附件及大网膜切除术＿肿瘤细胞减灭术＋盆腔淋巴结清扫术）。术后采用VAC（长春新碱，阿霉素，环磷酰胺）方案、BEP（博莱霉素或平阳霉素，足叶乙叉甙，顺铂或卡铂）方案或VBP（长春新碱，博莱霉素或平阳霉素，顺铂或卡铂）方案进行联合化疗。患者5年生存率根治性手术为44．4％，保守性手术为64．0％，差异无显著性。VAC方案与BEP（或VBP）方案的5年生存率分别为33．3％和77．3％（P＜0．01）。疗程数≥4患者的5年生存率为73．9％，明显高于疗程数＜4患者的38．9％（P＜0．05）。认为对于有生育要求的卵巢未成熟畸胎瘤患者，可仅行保守性手术，术后辅以联合化疗。BEP（或VBP）方案对患者长期生存率的改善明显优于VAC方案。术后化疗的疗程数与疗效有关，对长期生存率的影响较明显。     

Differential Regulation of RGS-2 by Constant and Oscillating PTH Concentrations

PTH has diverse effects on bone metabolism: anabolic when given intermittently, catabolic when given continuously. The cellular mechanisms underlying the varying target cell response are not clear yet. PTH induces RGS-2, a member of the Regulator of G-protein Signaling protein family, via cAMP/PKA, and inactivates PKC-mediated signaling. To investigate intracellular signaling pathways with different PTH concentration-time patterns, we treated UMR 106-01 osteoblast-like cells in a perfusion system. PTH was administered intermittently (4 min/h, 10⁻⁷ M) or continuously at an equivalent cumulative dose (6.6 × 10⁻⁹ M). cAMP was measured using radioimmunoassay, mRNA levels using real-time rtPCR and ribonuclease protection assay, and protein levels using Western immunoblotting. A single PTH pulse transiently increased cAMP levels by 2000% ± 1200%. In contrast to continuous PTH exposure, cAMP induction remained unchanged with intermittent PTH, ruling out desensitization of the PTH receptor. In continuously perfused cells, RGS-2 abundance was three to five times higher than in cells intermittently exposed to PTH for up to 12 h. MKP-1 and -3 were significantly less induced with pulsatile PTH; exposure-mode-dependent differences in MMP-13 and IGFBP-5 were small. Pulsatile but not continuous PTH administration prevents PTHrP receptor desensitization and accumulation of RGS-2 in osteoblasts, which should preserve PKC-dependent signaling.     

肝癌多药耐药产生与低糖环境的关系

目的探讨局部微环境低糖与肝细胞癌多药耐药性(MDR)产生的关系及影响机制。方法低糖培养HepG2细胞,应用流式细胞术Annexin V/PI法检测低糖培养的细胞在化疗药物5-氟脲嘧啶(5-Fu)作用后的凋亡情况,分别应用荧光定量聚合酶链反应(PCR)技术和Western blot技术检测低糖培养后HepG2细胞内多药耐药相关基因mdr1、MRP1、LRP的mRNA和蛋白水平的表达。结果在低糖环境下生长时间越长的HepG2细胞对5-Fu的抵抗越强,而且随着低糖培养时间的增加,5-Fu诱导的HepG2细胞的凋亡高峰延迟。低糖培养的HepG2细胞内多药耐药相关基因mdr1、MRP1、LRP在mRNA和蛋白水平的表达随低糖培养时间的延长而升高,以LRP的改变最为显著。结论肝癌生长微环境葡萄糖耐量不足也是肝癌产生MDR的原因之一。低糖可通过上调一组多药耐药相关基因的表达而诱导肝癌的多药耐药性。     

低位直肠癌的保肛手术

目的 探讨如何选择合适的保肛手术方法治疗低位直肠癌。方法 44例肿块下缘距肛缘5～7cm的低位直肠癌患者,对21例采用改良的结肠肛管吻合术(A组)、23例应用吻合器技术的前切除术(B组)进行治疗;并对两组疗效予以对比。结果 A组患者无远端直肠残端肿瘤残留,B组有2例。肿瘤远端直肠切除距离A组平均为(2.81±0.35)cm,B组(1.73±0.42)cm。两组比较,差异有显著性意义(t=9.083,P<0.001)。A组术后两年均无吻合口复发,B组有4例,B组吻合口复发率明显高于A组(x~2=4.234,P=0.04)。A组术后早期排便功能较差,但均能在半年内改善;B组术后排便功能良好。结论 低位直肠癌患者肿瘤下缘距肛缘5～7cm、肿瘤“T”分期为Ⅱ、Ⅲ期、术前指诊肿瘤可推动、肿瘤侵犯肠壁范围不到1周可行保肛手术。应根据患者体型、骨盆宽窄、肿瘤分化程度及其侵犯肠管的周径合理选择改良的结肠肛管吻合术或吻合器技术的前切除术。     

荷人卵巢癌严重联合免疫缺陷鼠腹腔移植模型的建立

目的 :建立荷人卵巢癌细胞株 SKOV3严重联合免疫缺陷小鼠 (SCID)动物模型。方法 :SCID鼠腹腔注射人 SKOV3卵巢癌细胞 1× 10 7个 /只 ,饲养在无特定病原体环境中 ,观察其生物学特性。结果 :实验组 3只 SCID小鼠腹腔内均见肿瘤生长 ,并形成以横膈、肠系膜、肝周、腹膜为主的转移灶 ,其中 1只肺部有镜下转移灶。移植后肿瘤细胞在形态、生长、分泌 CA12 5功能方面均与原细胞株保持一致。结论 :建立的荷人 SKOV3卵巢癌 SCID鼠腹腔移植模型能较好地模拟人卵巢癌腹腔播散的生物学特征。     

瘦蛋白的分子生物学特性研究进展

瘦蛋白 (leptin)是由肥胖基因编码、脂肪细胞分泌的一种分子量为 16kDa的蛋白质 ,是一种重要的代谢调节信号 ,对人和动物的采食量、能量代谢、繁殖性能等具有重要调节功能 ,对人肥胖病的治疗和改善动物的生产性能、胴体组成等有良好的潜在应用前景 .主要概述了瘦蛋白基因的克隆、转录、表达调控以及瘦蛋白受体与信号转导等分子生物学特性     

头颈部肿瘤CT灌注成像表现与微血管生成因子表达水平的相关性研究

目的研究头颈部肿瘤CT灌注与肿瘤血管生成因子（VEGF）的相关性。方法对85例共88个（恶性77个，良性11个）头颈部肿瘤术前行CT灌注检查。采用螺旋CT机自带软件绘制感兴趣区（ROI）的时间-密度曲线（TDC）并计算ROI强化峰值（PH）、达峰时间（PT）、平均通过时间（MTT），病灶相对强化峰值（RPH）和灌注量（PF）。其中35例切取与CT灌注靶层面相同的组织切片，行CD34、VEGF抗体免疫组织化学染色，分析肿瘤CT灌注成像表现与微血管密度（MVD）和VEGF表达的相关性。结果（1）头颈部肿瘤CT灌注成像TDC主要有3种类型，77个恶性肿瘤中53个（68．9％）表现为速升速降型；9个淋巴瘤中6个TDC表现为低平型曲线，与68个其他肿瘤中仅有9个为低平型曲线相比差异有统计学意义（P〈0．05）。（2）甲状腺癌呈高灌注，其PF（中位数为82．2ml·min^-1·100g^-1）与淋巴瘤PF（中位数为24．5ml·min^-1·100g^-1）、头颈鳞癌PF（中位数为23．8ml·min^-1·100g^-1）相比差异有统计学意义（P〈0．05）。（3）11个良性肿瘤的MVD均数为（44．7±3．4）条／高倍视野，24个恶性肿瘤的MVD为（49．6±14．8）条／高倍视野，良恶性肿瘤间差异无统计学意义（P〉0．05）；VEGF在恶性肿瘤呈强阳性者15个，弱阳性9个；在良性肿瘤呈强阳性1个，弱阳性10个，VEGF的表达强阳性率在良、恶性肿瘤差异有统计学意义（P〈0．01）。（4）MVD（中位数40．0）与PH（中位数26．9）、RPH（中位数14．5）和PF（中位数46．8）有明显相关性（r值分别为0．35、45．49和0．41），VEGF（中位数4．0）表达与MTT（中位数16．7）呈负相关（r=-0．41）。结论CT灌注成像TDC形态对头颈部肿瘤诊断及鉴别诊断有一定的帮助。MVD、VEGF与CT灌注相关，CT灌注成像可以反映肿瘤微循环情况。     

Synergistic effect of low dose Cyclosporine A and human interleukin 10 overexpression on acute rejection in rat lung allotransplantation

Objective: Electroporation mediated transfer of plasmid DNA into peripheral muscle results in high transfection efficiency. The aim of this study was to investigate the effect of gene transfer of human IL-10 (hIL-10) into the tibialis anterior muscle (MTA) in combination with low dose Cyclosporine A (CsA) on acute rejection of lung allografts in the rat. Methods: Lung allotransplantation was performed from male BN donor to male Fisher F344 rats. Gene transfer was achieved by intramuscular injection into the MTA of the recipient followed by electroporation (4×20 ms impulses at 200 V/cm) 24 h prior to the transplantation. Group A (n=5) received CsA (2.5 mg/kg bw ip) for 5 days post-transplant and group B (n=5) 2.5 μg of PCIK hIL-10 (plasmid expression vector containing human CMV immediate early gene promoter and enhancer) and a low dose CsA (2.5 mg/kg bw i.p.). Graft function was assessed by blood gas at day 5 after exclusion of the native lung. Animals were sacrificed and blood was drawn to measure serum hIL-10 levels (ELISA) and tissue was sampled for histological grading of rejection. Results: Local expression of hIL-10 was confirmed at the mRNA level by in situ hybridization. All group A control animals showed severe signs of rejection. At day 5 all grafts in group B showed good gas exchange mean PaO2 233±123 mmHg, vs 44±8 mmHg in group A. Histological examination revealed moderate to severe rejection in all animals in group A (IIIB, ISHLT) in contrast to low moderate rejection in group B (II–IIIA). hIL-10 serum levels on day 5 were 14±7 pg/ml in group B vs. 0 in group A. Conclusions: Electroporation mediated hIL-10 overexpression in a peripheral muscle of the recipient in combination with low dose CsA reduces acute rejection in this model of rat lung allotransplantation.     

性别差异对脓毒症大鼠肝脏Toll样受体4和髓样分化蛋白-2基因表达的影响

目的探讨性别差异对脓毒症大鼠肝脏组织Toll样受体4(TLR4)和髓样分化蛋白- 2(MD-2)基因表达的影响。方法以脂多糖(LPS)按5 mg/kg体重由大鼠腹腔注射制作脓毒症动物模型,注射后2 h留取肝脏组织检测TLR4、MD-2和肿瘤坏死因子-α(TNF-α)基因表达,同时测定各组大鼠血浆中丙氨酸氨基转移酶(ALT)及雌二醇含量。结果正常雌雄性大鼠肝脏组织均可表达少量TLR4、MD-2、TNF-α基因,其中雌性组分别为0．175±0．034、0．211±0．044、0．201±0．068; 雄性组分别为0．205±0．061、0．243±0．049、0．243±0．063,两组数据差异无统计学意义(P> 0．05),但LPS刺激后雌性大鼠肝脏组织上述指标分别为0．615±0．089、0．708±0．181、0．730± 0．118,血浆中ALT含量为(81．07±10．72)U／L;雄性组分别为0．723±0．091、1．123±0．272、 0．881±0．156,ALT含量为(106．39±14．21)U／L,雌性组各项指标均明显低于雄性大鼠(P< 0．05)。相关分析表明雌性及雄性脓毒症大鼠肝脏组织TLR4及TNF-α基因表达与相应性别大鼠血浆中雌二醇含量呈显著负相关(P<0．05)。结论 LPS刺激后大鼠肝脏组织TLR4、MD-2及 TNF-α基因表达存在性别差异,内源性雌激素的作用可能导致雌性脓毒症大鼠肝脏组织损伤较雄性轻。