河南省汉族人群人类白细胞抗原DRB1基因多态性与系统性硬化病及其肾损害的关联研究

目的 探讨河南省汉族人群人类白细胞抗原( HLA )-DRB1基因多态性与系统性硬化病(SSc)及其肾损害的关联性.方法 采用巢式聚合酶链反应-直接测序分型(PCR-SBT)对81例SSc患者[其中59例合并硬皮病肾损害(SRD)]及90名健康对照的HLA-DRB1基因型进行研究分析.组间差异比较采用x2检验或四格表确切概率法,多因素之间的分析采用Logistic回归分析.结果 共发现33种HLA-DRB1等位基因,SSc患者中有27种,SRD中有22种.SSc组HLA-DRB1*040501、*110101、*150201等位基因分布频率(分别为8.64％、11.11％、8.02％)高于健康对照组(分别为1.67％、4.44％、2.22％),Logistic回归分析显示HLA-DRB1* 040501(P=0.010,OR=5.839,95％CI:1.518～22.460)、*110101(P=0.019,OR =3.060,95％CI:1.204～7.772)、*150201(P=0.010,OR=4.780,95％CI:1.444～15.821)分别与SSc的发病独立相关.SRD患者组中HLA-DRB1* 040501、*150201等位基因分布频率(分别为9.32％、7.63％)较对照组(分别为1.67％、2.22％)升高,Logistic回归分析显示HLA-DRB1* 040501(P=0.008,OR=6.363,95％CI:1.614～25.087)、*150201(P=0.030,OR =4.043,95％CI:1.147～14.243)分别与SRD的发病独立相关.结论 在河南汉族人群中HLA-DRB1* 040501、*110101、*150201可能是SSc发病的易感基因;而HLA-DRB1* 040501、*150201可能是SRD发病的易感基因.     

艾灸预处理对大鼠应激性胃黏膜损伤增殖修复相关因子的影响

目的:观察艾灸预处理对应激性胃黏膜损伤大鼠血清和胃黏膜表皮生长因子(EGF)、转化生长因子-α(TGF-α)、胃黏膜三叶因子家族-1(TFF1)、增殖细胞核抗原(PCNA)的影响,探讨艾灸预处理促进胃黏膜损伤增殖修复的作用机制.方法:将48只健康SD大鼠随机分为4组,即空白组、模型组、艾灸穴位组、艾灸非穴组.束缚冷应激法制作大鼠应激性胃黏膜损伤模型.造模之前,艾灸组选取足三里、中脘、脾俞和胃俞等穴位行艾灸预处理8d,艾灸非穴组选取非穴对照点进行预处理.以Guth法计算胃黏膜损伤指数,光镜下观察大鼠胃黏膜组织学改变,放射免疫法测定血清EGF与TGF-α的含量,酶免法检测胃黏膜组织中EGF、TGF-α、TFF1和PCNA的含量.结果:与模型组和艾灸非穴组比较,艾灸足三里、中脘等穴位可使应激性胃黏膜损伤大鼠胃黏膜损伤指数明显下降(14.667±5.710vs27.250±7.448,24.750±7.300,P<0.01),血清EGF、TGF-α含量升高(2.167±0.756vs1.147±0.983,1.358±0.962,P<0.05;11.170±1.315vs4.585±0.720vs5.118±0.659,P<0.01),胃黏膜EGF、TGF-α和PCNA含量升高(343.560±27.644vs269.610±45.119,279.590±58.890,P<0.05;147.470±17.784vs115.530±24.319,116.620±14.908,P<0.01;191.910±37.262vs154.580±18.910,152.450±20.333,P<0.05);与模型组比较,艾灸穴位组胃黏膜TFF1含量明显升高(4.573±0.121vs3.654±0.507,P<0.05).结论:艾灸足三里、中脘等穴位预处理可减轻束缚水浸应激所造成大鼠胃黏膜损伤、促进胃黏膜损伤组织增殖修复,可能是通过上调胃黏膜损伤增殖修复相关因子(EGF、TGF-α、TFF1和PCNA)而达到其促胃黏膜损伤修复的作用.     

Pharmacokinetic properties of IB1001, an investigational recombinant factor IX,in patients with haemophilia B: repeat pharmacokinetic evaluation and sialylation analysis

IB1001 trenacog alfa is an investigational recombinant factor IX (FIX) for the treatment and prevention of bleeding in individuals with haemophilia B. To compare the pharmacokinetics (PK) of IB1001 with nonacog alfa in individuals with haemophilia B and to assess the relationship between sialylation and PK of IB1001 (NCT00768287). A randomized, double‐blind, non‐inferiority, cross‐over study conducted in participants aged ≥12 years weighing ≥40 kg, with severe or moderately severe haemophilia B (FIX activity ≤2 IU dL ^?1). PK parameters were derived using observed FIX concentration levels and actual PK sampling times, and repeated in a subset of participants who had received IB1001 prophylaxis for 4–18 months. A retrospective analysis was conducted in subgroups according to the sialylation levels of IB1001 (50.8, 57.8–59.0%, or 71.7%). In the 32 adolescent and adult males evaluated, there were no clinically meaningful differences in PK parameters between those receiving IB1001 75 IU kg^?1 or nonacog alfa. The lower limit of the one‐sided 95% confidence interval for the ratio of AUC_0‐t and AUC_0‐∞ (IB1001/nonacog alfa) was 0.90, establishing non‐inferiority. Terminal phase half‐lives were similar (29.7 ± 18.2 h for IB1001 and 33.4 ± 21.2 h for nonacog alfa). The PK results were stable for up to 18 months of IB1001 exposure; the impact of sialylation levels was not clinically meaningful. There were no clinically meaningful PK differences between IB1001 and nonacog alfa. IB1001 was well tolerated and without safety concerns. The non‐inferiority of IB1001 to nonacog alfa supports IB1001 becoming a useful alternative recombinant agent for the management of haemophilia B.     

Prediction of response to entecavir therapy in patients with HBeAg-positive chronic hepatitis B based on on-treatment HBsAg, HBeAg and HBV DNA levels

Summary. Quantitative hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) assays are emerging as effective tools of on‐treatment predictors of response to antiviral agents, in addition to monitoring serum HBV DNA levels. However, the dynamic relationship between quantitative HBsAg, as well as HBeAg and HBV DNA, and the predictability of subsequent clinical outcomes during entecavir (ETV) therapy remain unclear. Eighty‐two patients with HBeAg‐positive chronic hepatitis B (CHB) received ETV therapy for ≥3 years. Virologic response (VR) after 3 years of ETV therapy was achieved in 73 (89.0%) patients. Among baseline and on‐treatment factors, on‐treatment HBV DNA levels performed better with respect to the prediction of response than HBsAg and HBeAg levels. Especially, the performance of absolute values of HBV DNA with respect to response was superior to HBV DNA decline from the baseline. The best predictive value was an absolute HBV DNA level of 2.3 log₁₀ IU/mL at month 6 (areas under the curve [AUROC], 0.977; 95% CI, 0.940–1.000; P < 0.001). HBeAg seroconversion after 3 years of therapy was achieved in 26 (31.7%) patients. On‐treatment HBeAg levels performed better with respect to the prediction of seroconversion than HBsAg and HBV DNA levels. The best cut‐off value for the HBeAg level at month 12 for the prediction of seroconversion was 0.62 log₁₀ PEIU/mL. Although the HBsAg level at baseline is often used to predict the antiviral potency of entecavir, on‐treatment HBV DNA and HBeAg levels are more helpful for prediction of subsequent clinical outcomes in HBeAg‐positive CHB patients with entecavir treatment.     

Combination of hepatitis B viral antigens and DNA for prediction of relapse after discontinuation of nucleos(t)ide analogs in patients with chronic hepatitis B

Aim: The factors associated with hepatitis recurrence after discontinuation of nucleos(t)ide analogs (NAs) in patients with chronic hepatitis B were analyzed to predict the risk of relapse more accurately. Methods: A total of 126 patients who discontinued NA therapy were recruited retrospectively. The clinical conditions of a successful discontinuation were set as alanine aminotransferase (ALT) below 30 IU/L and serum hepatitis B virus (HBV) DNA below 4.0 log copies/mL. Results: Relapse of hepatitis B were judged to occur when maximal serum ALT became higher than 79 IU/L or when maximal serum HBV DNA surpassed 5.7 log copies/mL following NA discontinuation since these values corresponded with mean values of ALT (30 IU/L) and HBV DNA (4.0 log copies/mL), respectively. At least 90% of patients with either detectable hepatitis B e antigen or serum HBV DNA higher than 3.0 log copies/mL at the time of NA discontinuation relapsed within one year. In the remaining patients, higher levels of both hepatitis B surface and core‐related antigens at the time of discontinuation, as well as a shorter course of NA treatment, were significantly associated with relapse by multivariate analysis. Conclusions: It appears that negative results for hepatitis B e antigen and serum HBV DNA lower than 3.0 log copies/mL are essential for successful NA discontinuation, which may be attained by a longer treatment period. Levels of hepatitis B surface and core‐related antigens are also significant factors independently associated with relapse of hepatitis.     

Sunitinib对支气管哮喘气道重塑小鼠模型Erk通路和cyclin D1表达的影响

目的 探讨sunitinib对支气管哮喘(简称哮喘)气道重塑的干预作用及可能的作用机制.方法 18只BALB/c小鼠随机分为对照组、哮喘组以及sunitinib组,每组6只;以卵清蛋白(OVA)致敏、激发,建立慢性哮喘气道重塑模型.Sunitinib组每次雾化吸人前半小时给予sunitinib(40 mg/kg)灌胃给药.OVA末次激发结束后24 h处死小鼠,HE染色观察气道炎症及形态学改变,采用ELISA法检测支气管肺泡灌洗液(BALF)中IL-4、IL-13和血清总IgE的表达,免疫组织化学法观察肺组织增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、α平滑肌肌动蛋白(a smooth muscle actin,α SMA)表达水平.蛋白印记法(Western blot)测定细胞外信号调节蛋白激酶(extraeellular signal-regulated kinase,Erk)蛋白磷酸化水平及细胞周期蛋白D1(cyclin D1)的表达.结果 HE染色示哮喘组小鼠黏膜下层和平滑肌层增厚,管腔狭窄,大量炎性细胞浸润,sunitinib组上述改变较哮喘组为轻;sunitinib干预后哮喘小鼠BALF中Th2细胞因子IL4、IL-13和血清总IgE以及肺组织羟脯氨酸含量显著降低(P＜0.01).哮喘组小鼠气道PCNA阳性细胞百分比、α-SMA表达及肺组织Erk磷酸化水平、cyclin D1蛋白表达较对照组明显升高(P＜0.01),sunitinib干预后其表达均降低(P＜0.01).结论 Sunitinib可能通过抑制Erk途径影响cyclin D1的表达,抑制了慢性哮喘模型中气道平滑肌的增殖,发挥抗气道重塑作用.     

日本血吸虫可溶性虫卵抗原免疫小鼠CD11c~+CD8_α~-DC对过敏性哮喘的抑制作用

目的研究日本血吸虫可溶性虫卵抗原(SEA)免疫小鼠CD11c+CD8α-树突状细胞(DC)亚群对卵清白蛋白(OVA)诱发的过敏性哮喘的抑制作用。方法 BALB/c小鼠经腹腔及足垫注射SEA 50μg/只,每周1次,共4次。用抗体包被的免疫磁珠分离小鼠脾CD11c+CD8α+DC与CD11c+CD8α-DC亚群。另取18只BALB/c小鼠,随机分为4组,A组为健康对照组,B组为OVA致敏单纯哮喘组,C组为过继转移SEA免疫CD11c+CD8α+DC组,D组为过继转移SEA免疫CD11c+CD8α-DC组。C、D组小鼠分别经尾静脉过继转移5×105个CD11c+CD8α+DC和5×105个CD11c+CD8α-DC,1h后B、C、D组小鼠同时用OVA诱发哮喘,4周后剖杀,取肺组织,做病理切片,经苏木素-伊红(HE)染色后,光镜下观察肺部炎症变化。结果 A组小鼠肺组织无炎症反应;B组小鼠肺组织炎症反应广泛且严重,在支气管及肺泡周围有大量炎性细胞浸润;C组小鼠肺组织炎症反应仍较明显;D组小鼠肺组织炎症反应较B、C组显著减轻,仅有少量炎细胞浸润。4组小鼠按照Underwood标准进行病理评分,总分分别为0、14.00±1.00、12.33±0.58和7.20±1.30,差异有统计学意义(P<0.05)。结论日本血吸虫SEA免疫小鼠CD11c+CD8α-DC亚群对过敏性哮喘有抑制作用。     

良性前列腺增生与血生化代谢指标的关系分析

目的 探讨BPH与血生化代谢指标的关系. 方法 回顾性分析2008年1月至2010年10月收治的73例术后病理确诊为BPH的患者资料.年龄55 ～86岁,平均70岁.其中术前3个月内未服用影响血糖、血脂等代谢水平药物的患者65例,分析患者前列腺总体积、年均增长率与血糖、血脂、血前列腺特异性抗原(PSA)之间的关系.73例患者按有无合并代谢综合征(MS)分为两组,比较两组间前列腺总体积、年均增长率、血PSA等临床指标. 结果 65例BPH患者前列腺总体积为(58.76±25.96) ml,年均增长率为(1.32±0.89)ml,空腹血糖为(4.99±0.73) mmol/L,高密度脂蛋白为(1.36±0.39) mmol/L,血tPSA值为(5.99 ±8.30)μg/L.按患者前列腺总体积分组后统计分析显示空腹血糖(F=4.63,P=0.0060)、血tPSA值(F=4.83,P=0.0044)、体积年均增长率(F=62.98,P=0.0001)随前列腺总体积增加而升高,而高密度脂蛋白降低(F=5.32,P=0.0025).对前列腺总体积控制后仍显示前列腺总体积与空腹血糖呈正相关(r =0.48,P=0.0005),与高密度脂蛋白呈负相关(r=-0.26,P=0.0353).73例患者中合并MS者20例,血tPSA(6.52±5.28) μg/L、前列腺总体积(79.41±38.24) ml、年均增长率(1.86±1.14)ml,无合并MS者分别为(3.95±3.77) μg/L、(54.98±20.94)ml及(1.23 ±0.80) ml,组间差异均有统计学意义(t=2.29,P=0.0253;t =2.65,P=0.0147;t=2.22,P=0.0360). 结论 BPH患者前列腺总体积及年均增长率、血tPSA与血糖、血脂等代谢水平有关,与空腹血糖呈正相关,与高密度脂蛋白呈负相关.合并MS者前列腺临床进展特性更明显.     

血管内皮生长因子和骨形态发生蛋白7在细胞分化中的协同作用

目的：研究rhVEGF和rhBMP-7对成骨细胞分化能力的影响.方法：取大鼠颅盖骨组织块,采用改良组织块混合酶消化法培养成骨细胞,将不同浓度的rhVEGF和rhBMP-7加入第四代成骨细胞的培养基继续进行细胞培养,用碱性磷酸酶试剂盒检测定细胞在第1、3、5天的磷酸酶的活性,分析不同浓度的rhVEGF和rhBMP-7对成骨细胞分化的影响.结果：rhVEGF和rhBMP-7能促进成骨细胞的分化,其中,3.125ng/mlrhVEGF和50.000ng/ml rhBMP-7单独或者两者联合作用并于第3天时,对成骨细胞碱性磷酸酶活性的影响最明显.结论：一定剂量的rhVEGF和rhBMP-7可明显促进成骨细胞的分化.