慢性乙肝患者生长激素-胰岛素样生长因子轴的变化及其临床意义

目的探讨慢性乙型肝炎(慢性乙肝)和肝硬化患者生长激素-胰岛素样生长因子轴(growth hormone-insulin-like growth factor axis,GH-IGF)的变化及其临床意义。方法采用酶联免疫吸附法检测39例慢性乙肝、37例乙肝肝硬化及12例健康献血者血清胰岛素样生长因子1(insulin-like growth factor-1,IGF-1)和胰岛素样生长因子结合蛋白-3(insulin-like growth factor binding protein-3,IGFBP-3),采用RIA法检测血清生长激素(growthhormone,GH)。其中26例肝组织标本行IGF-1的免疫组织化学染色,并采用多媒体彩色图文分析系统进行图像定量分析。结果①慢性乙肝和肝硬化患者血清GH水平较正常对照组显著升高(P<0.05),而血清IGF-1和IGFBP-3水平明显下降(P<0.05);②慢性肝炎组随着炎症活动度的增加血清GH水平升高,血清IGF-1和IGFBP-3水平逐渐降低(P<0.05),肝硬化组随着肝功能代偿能力的减退血清GH水平升高,血清IGF-1和IGFBP-3水平明显下降(P<0.05);③G1~G4组肝组织IGF-1的表达较G0组增加(P<0.05),但G1~G4组之间无显著性差异(P>0.05)。结论慢性乙肝和肝硬化患者GH-IGF轴发生显著性变化,存在GH抵抗现象。血清GHI、GF-1I、GFBP-3水平在一定程度上反映肝细胞的炎症坏死程度及肝脏合成储备功能。     

可溶性B、T淋巴细胞衰减因子及其配体在单纯疱疹性角膜基质炎小鼠体内的异常表达

目的 研究B、T淋巴细胞衰减因子(B and T lymphocyte attenuator,BTLA)及其配体疱疹病毒侵入介体(herpes virus entry mediator,HVEM)在单纯疱疹性角膜基质炎(herpetic stromal keratitis,HSK)小鼠角膜组织中及外周血CD4+T细胞上的表达水平,探讨共抑制信号BTLA-HVEM是否参与了CD4+T细胞介导的HSK免疫病理反应.方法 将106 PFU的单纯疱疹病毒Ⅰ型(herpes simplex virus type 1,HSV-1)KOS毒株接种于BALB/c鼠的角膜上建立HSK动物模型,分别于角膜接种病毒前(0 d),接种病毒后的第3、7、10、14、21天,用毛细管取小鼠的左眼眼眶静脉窦血1 mL,分离淋巴细胞,行荧光抗体染色,用流式细胞仪检测CD3+ CD4+ BTLA+T细胞和CD3+ CD4+ HVEM+T细胞阳性率;在裂隙灯显微镜下观察小鼠角膜变化;免疫组织化学方法检测BTLA蛋白及HVEM蛋白在角膜组织中的表达.结果 BALB/c鼠的角膜接种HSV-1后的1～5d,角膜擦拭液中均检测出HSV-1复制,表明小鼠感染了单纯疱疹病毒.裂隙灯显微镜观察显示:角膜接种HSV-1后第3天,所有小鼠均患了急性上皮性角膜炎,并于感染后1周内痊愈,自病毒接种后第8天起,小鼠出现角膜基质炎的改变,表现为角膜基质呈灰白色混浊,角膜基质混浊于病毒接种后的第10天达到高峰,持续至第14天后逐渐减轻.流式细胞仪检测显示,小鼠外周血淋巴细胞中CD3+CD4+BTLA+T细胞和CD3+ CD4+ HVEM+T细胞的阳性率,在角膜接种病毒前(0 d)分别为(3.15±0.60)％和(9.84±1.06)％,在角膜接种病毒后第10天(HSK疾病程度最严重时)分别增加到(20.47±3.15)％和(45.18±3.90)％(与0d相比差异均有显著统计学意义,均为P＜0.01).免疫组织化学方法检查HSK小鼠角膜组织中BTLA和HVEM的蛋白表达结果一致:HSK临床表现最严重时,即病毒接种后第10天时,BTLA蛋白和HVEM蛋白在角膜组织中表达最强,主要表达于角膜基质层内浸润的炎性细胞上,角膜上皮层和内皮层也有表达.结论 在HSK小鼠模型中,BTLA及其配体HVEM蛋白在角膜组织中及外周血CD4+T细胞上表达明显增强,共抑制信号BTLA-HVEM参与了CD4+T细胞介导的HSK的免疫病理过程.     

Effects of salvianolic acid B on in vitro growth inhibition and apoptosis induction of retinoblastoma cells

AIM: To observe the effects of salvianolic acid B (SalB) on in vitro growth inhibition and apoptosis induction of retinoblastoma HXO-RB44 cells. METHODS: The effects of SalB on the HXO-RB44 cells proliferation in vitro were observed by MTT colorimetric method. The morphological changes of apoptosis before and after the treatment of SalB were observed by Hoechst 33258 fluorescent staining method. Apoptosis rate and cell cycle changes of HXO-RB44 cells were detected by flow cytometer at 48 hours after treated by SalB. The expression changes of Caspase-3 protein in HXO-RB44 cells were detected by Western Blot. RESULTS: SalB significantly inhibited the growth of HXO-RB44 cells, while the inhibition was in a concentration-and time-dependent manner. The results of fluorescent staining method indicated that HXO-RB44 cells showed significant phenomenon of apoptosis including karyorrhexis, fragmentation and the formation of apoptotic bodies, etc. after 24, 48 and 72 hours co-culturing of SalB and HXO-RB44 cells. The results of flow cytometer showed that the apoptosis rate and the proportion of cells in S phase were gradually increased at 48 hours and 72 hours after treated by different concentrations of SalB. Western Blot strip showed that the expression of Caspase-3 protein in HXO-RB44 cells was gradually increased with the increase of the concentration of SalB. CONCLUSION: SalB can significantly affect on HXO-RB44 cells growth inhibition and apoptosis induction which may be achieved through the up-regulation of Caspase-3 expression and the induction of cell cycle arrest.     

131I治疗后Graves眼病血清TNF-α、sTNFRⅡ和sICAM-1的相关性研究

目的探讨G raves病眼病(GO)伴有甲状腺功能亢进症(甲亢)患者经131I治疗后,血清肿瘤坏死因子-α(TNF-α)、可溶性肿瘤坏死因子受体Ⅱ(sTNFRⅡ)和可溶性细胞间粘附分子-1(sICAM-1)的相关性。方法 131I治疗6～12个月间,根据"NOSPECS眼部病变分级"分为无GO组14例、轻度GO组35例和中度GO组35例,按"CAS评分"分为GO静止组32例和GO活动组38例。设对照组29例。采用ELISA方法检测各组患者血清sI-CAM-1、TNF-α和sTNFRⅡ水平。结果 GO活动组血清sICAM-1(1211.9±258.6ng/m l)水平高于对照组(697.8±291.0 ng/m l)、无GO组和GO静止组(797.6±250.0 ng/m l、807.9±330.2ng/m l,P<0.01),GO静止组和GO活动组血清TNF-α(378.1±167.9ng/L、391.6±150.7ng/L)水平均高于无GO组和对照组(P<0.01);无GO组、GO静止组和GO活动组血清sTNFRⅡ(730.0±173.3ng/L、1055.1±422.9 ng/L、960.0±435.4 ng/L)水平均高于对照组(P<0.01)。轻度GO组、中度GO组血清sICAM-1(1032.4±232.3 ng/m l、407.0±158.1 ng/L)和TNF-α(1021.9±448.9 ng/m l、375.2±157.8 ng/L)水平均高于无GO组和对照组(P<0.05);而无GO组、轻度GO组和中度GO组血清sTNFRⅡ(730.0±173.3 ng/m l、1154.4±422.9 ng/L、922.1±440.6 ng/L)水平均高于对照组(P<0.05)。全部患者血清sICAM-1分别与TNF-α、sTNFRⅡ呈正相关,(r=0.377,P=0.001,r=0.434,P=0.001),TNF-α和sTNFRⅡ之间呈正相关(r=0.476,P<0.001)。结论 GO患者131I治疗的早期,通过血清sICAM-1、TNF-α和sTNFRⅡ的表达对活动性突眼、眼眶内免疫炎性反应,纤维组织变化和甲亢缓解进行综合评价,并可结合CAS评分了解GO患者预后、甲状腺及眼眶的免疫学及细胞凋亡的状态,指导有针对性治疗。     

Fetal stroke and cerebrovascular disease: Advances in understanding from lenticulostriate and venous imaging,alloimmune thrombocytopaenia and monochorionic twins

Fetal stroke is an important cause of cerebral palsy but is difficult to diagnose unless imaging is undertaken in pregnancies at risk because of known maternal or fetal disorders. Fetal ultrasound or magnetic resonance imaging may show haemorrhage or ischaemic lesions including multicystic encephalomalacia and focal porencephaly. Serial imaging has shown the development of malformations including schizencephaly and polymicrogyra after ischaemic and haemorrhagic stroke. Recognised causes of haemorrhagic fetal stroke include alloimmune and autoimmune thrombocytopaenia, maternal and fetal clotting disorders and trauma but these are relatively rare. It is likely that a significant proportion of periventricular and intraventricular haemorrhages are of venous origin. Recent evidence highlights the importance of arterial endothelial dysfunction, rather than thrombocytopaenia, in the intraparenchymal haemorrhage of alloimmune thrombocytopaenia. In the context of placental anastomoses, monochorionic diamniotic twins are at risk of twin twin transfusion syndrome (TTTS), or partial forms including Twin Oligohydramnios Polyhydramnios Sequence (TOPS), differences in estimated weight (selective Intrauterine growth Retardation; sIUGR), or in fetal haemoglobin (Twin Anaemia Polycythaemia Sequence; TAPS). There is a very wide range of ischaemic and haemorrhagic injury in a focal as well as a global distribution. Acute twin twin transfusion may account for intraventricular haemorrhage in recipients and periventricular leukomalacia in donors but there are additional risk factors for focal embolism and cerebrovascular disease. The recipient has circulatory overload, with effects on systemic and pulmonary circulations which probably lead to systemic and pulmonary hypertension and even right ventricular outflow tract obstruction as well as the polycythaemia which is a risk factor for thrombosis and vasculopathy. The donor is hypovolaemic and has a reticulocytosis in response to the anaemia while maternal hypertension and diabetes may influence stroke risk. Understanding of the mechanisms, including the role of vasculopathy, in well studied conditions such as alloimmune thrombocytopaenia and monochorionic diamniotic twinning may lead to reduction of the burden of antenatally sustained cerebral palsy.     

小儿病毒性心肌炎与柯萨奇B组病毒抗体的关系探讨

目的探讨柯萨奇 Ｂ组病毒抗体ＩｇＭ测定在心肌炎病原学诊断和临床诊断中的意义。方法１９９６～１９９８年儿科住院病人７０例,分为心肌炎组２０例,疑似心肌炎组２５例,对照组２５例。用间接酶联免疫吸附法（ＥＬＩＳＡ）测定柯萨奇Ｂ组病毒抗体ＩｇＭ和ＩｇＧ。结果柯萨奇Ｂ组病毒抗体ＩｇＭ阳性心肌炎组１１例（５５％）,疑似心肌炎组１１例（４４％）,对照组１１例（４４％）,经统计学处理,其差别无显著性意义。结论用ＥＬＩＳＡ法测定柯萨奇Ｂ组病毒抗体ＩｇＭ,阳性提示有柯萨奇Ｂ组病毒感染,对心肌炎患儿可以作出病因和病原学诊断,但不能以此作为临床诊断心肌炎的依据。     

柯萨奇B组病毒感染与小儿哮喘发作关系的探讨

目的　探讨柯萨奇 Ｂ 组病毒（ Ｃ Ｖ Ｂ） 感染与哮喘发作的关系。方法　观察急性发作期哮喘患儿１０２ 例、上呼吸道感染１８２ 例、非感染性疾病患儿７１ 例、健康儿童３０ 例。采用 Ｅ Ｌ Ｉ Ｓ Ａ 法测定静脉血 Ｃ Ｖ Ｂ 抗原（ Ｃ Ｖ Ｂ－ Ａｇ）和 Ｉｇ Ｍ 抗体（ Ｃ Ｖ Ｂ－ Ｉｇ Ｍ） 、免疫组化法测定 Ｔ 细胞亚群和［３ Ｈ］ － Ｔｄ Ｒ 标记法测定 Ｎ Ｋ 细胞活性。结果　①哮喘患儿 Ｃ Ｖ Ｂ 感染率为５７８ ％ （５９／１０２） ;②哮喘患儿 Ｃ Ｄ８ 显著升高; Ｃ Ｄ３ 、 Ｃ Ｄ４／ Ｃ Ｄ８ 及 Ｎ Ｋ 细胞活性均明显下降（ Ｐ ＜００１） ;而 Ｃ Ｖ Ｂ 阳性哮喘患儿 Ｎ Ｋ 细胞活性又低于非 Ｃ Ｖ Ｂ 阳性哮喘儿（ Ｐ ＜ ００５） , Ｃ Ｄ８ 明显高于非 Ｃ Ｖ Ｂ 阳性哮喘儿（ Ｐ ＜ ００１） 。结论　 Ｃ Ｖ Ｂ 感染与哮喘发作有密切关系。     

甘草酸二铵治疗肝纤维化的动态观察

目的 :观察甘草酸二铵治疗肝纤维化的疗效。方法 :2 0 0例 [男性 135例 ,女性 6 5例 ,年龄 (37±s 2 1)a]慢性肝炎病人分为A组 4 5例 ,B组 15 5例。A组用门冬氨酸钾镁 2 0mL ,B组用甘草酸二铵 30 0mg ,均加入 5 %葡萄糖注射液 5 0 0mL ,ivgtt ,qd ,连用 6wk后改为口服药治疗 8wk。观察肝功能、血清透明质酸 (HA)、Ⅲ型前胶原 (PCⅢ )及肝组织学变化。结果 :治疗 6wk时 ,B组的ALT复常率为 74 .2 % ,明显优于A组 (4 6 % ) ,P <0 .0 1。治疗结束后 ,B组的肝纤维化程度明显减轻 ,与A组相比 ,HA和PCⅢ水平明显降低。结论 :甘草酸二铵不但有良好的改善肝功能作用 ,还具有抗肝纤维化作用     

长期应用恩替卡韦治疗的慢性乙型肝炎患者耐药发生情况及原因分析:8年随访研究

目的:了解慢性乙型肝炎患者长期应用恩替卡韦(ETV)治疗的耐药发生情况并探讨耐药原因。方法:对2003-2007年3月参加ETV剂量和疗效的随机双盲Ⅱ期临床试验、ETV与拉米夫定(LAM)治疗慢性乙型肝炎的随机双盲对照Ⅲ期临床试验、ETV治疗LAM失效的慢性乙型肝炎Ⅲ期临床试验及其后续ETV开放治疗临床试验的63例慢性乙型肝炎患者继续给予治疗和随访,计算截至2011年3月的累积耐药发生率,对临床耐药患者进行乙型肝炎病毒(HBV)逆转录酶区(RT)序列突变检测,并分析耐药原因。结果:63例患者中3例治疗2年后失访;1例治疗2.5年时转其他中心治疗;其他59例中16例用药4～6年,43例用药7～8年,随访4～8年,中位随访时间7年。13例患者发生临床耐药,累积耐药发生率为20.5%。9例耐药患者进行了HBV RT序列突变检测,其中5例检测出LAM耐药位点L180M和M204V突变和1～2个ETV耐药位点突变,符合ETV耐药特点;3例仅检出LAM耐药位点突变;1例检出2个ETV耐药位点突变和ADF耐药位点A181T突变。LAM经治、治疗前高病毒载量、治疗6个月病毒学应答不佳、治疗依从性差可能与耐药的发生有关。结论:长期应用恩替卡韦治疗慢性乙型肝炎耐药率较低。对治疗初期应答不佳患者,加强监测和提高治疗依从性可减少耐药的发生。     

恩替卡韦联合六味五灵片治疗乙型肝炎病毒e抗原阳性慢性乙型肝炎的临床观察

目的探讨恩替韦卡（ETV）联合六味五灵片治疗乙型肝炎病毒e抗原阳性慢性乙型肝炎的疗效和安全性。方法将124例乙型肝炎病毒e抗原阳性慢性乙型肝炎患者采用随机数字表法分为两组,分别采用ETV联合六味五灵片治疗和单纯ETV治疗,比较两组肝功能及病毒复制指标。结果观察组有效率91．94％,对照组有效率82．26％,两组差异有统计学意义（Ridit=32．251,P〈0．05）;治疗24周、48周,观察组HBVDNA低于对照组,差异有统计学意义（t=2．365、4．595,均P〈0．05）;治疗后,观察组ALT、AST下降程度显著大于对照组（t=78．512、62．145,均P〈0．05）;治疗后两组ALT复常率、总有效率和反跳率差异均有统计学意义（x^2=25．621、46．241、11．512,均P〈0．05）,HBVDNA和HBeAg阳性低于检测下限例数差异具有统计学意义（x^2=17．265、21．264,均P〈0．05）。结论恩替韦卡联合六味五灵片治疗乙型肝炎病毒e抗原阳性慢性乙型肝炎能够减轻肝脏纤维化．并具有明显解毒和抗病毒作用。