慢性乙肝患者NK,ADCC活性及其对IFN—α,IFN—γ的反应

本文用Hela细胞乳酸脱氢酶释放法测定了42例慢性乙肝病人外周血NK、ADCC活性及其对IFN-α100U/ml、500U/ml及IFN-γ500U/ml预处理4小时后的反应。结果显示,病人PBL NK,ADCC活性与正常对照有显著差别,经IFNs处理后,其NK或/和ADCC活性均有不同程度的升高反应,从而提示了IFNs在慢性肝病治疗中的重要作用。     

Inhibition of MAP kinase activity moderates changes in expression and function of Cx32 but not claudin-1 during DNA synthesis in primary cultures of rat hepatocytes

The signal transduction pathways and activation of the MAP kinase or PI3 kinase signaling cascade regulate a variety of cellular processes, including proliferation and differentiation in hepatocytes. To elucidate the mechanisms of signal transmission required for the regulation of gap and tight junctions during DNA synthesis in rat hepatocytes, we determined changes of expression and function of gap and tight junctions of cells grown in primary culture, using inhibitors of signaling pathways for MAP kinase (PD98059) and PI3 kinase (LY294002). During the stimulation of DNA synthesis induced by epidermal growth factor (EGF), immunoreactivity and mRNAs of gap junction protein Cx32 and of tight junction protein claudin-1 markedly decreased with reduction of gap junctional intercellular communication (GJIC) and the fence function of tight junctions. In Western blots, whole-cell lysate of claudin-1 protein decreased and phosphorylated Cx32 protein in the insoluble fraction of Triton X-100 increased during the stimulation of DNA synthesis. During reinhibition of DNA synthesis, the changes of Cx32 and claudin-1 returned to control levels, as did both functions. In treatment with the inhibitors before DNA synthesis, PD98059 inhibited the changes of expression and function of Cx32, but not claudin-1, without inhibition of cell growth, whereas LY294002 completely inhibited cell growth. These findings indicate that the PI3 kinase pathway rather than the MAP kinase pathway plays an important role for EGF-induced proliferation of rat hepatocytes, and that changes of Cx32 in hepatocytes during the stimulation of DNA synthesis may be in part controlled through MAP kinase. Furthermore, Cx32, but not claudin-1, protein may be a target of activated MAP kinase in hepatocytes.     

34例鹿角形肾结石手术取石的疗效观察

目的探讨肾下极肾盂肾盏联合切开术治疗复杂性鹿角形肾结石的疗效。方法回顾性分析采用肾下极肾盂肾盏联合切开术式治疗复杂性鹿角形肾结石34例临床资料。手术方法:分离肾窦内肾盂后,用1-0肠线在肾后唇中下1/3连接处紧贴肾盂肾盏表面缝合肾实质2针,中间尖刀切开肾实质,然后弧形切开肾盂及肾下盏,用神经剥离子将结石与肾盂肾盏粘膜粘连分离后,取石钳夹住结石轻轻撬出。盏颈狭窄者必要时先用气压弹道碎石器将其在分支处击断。4-0肠线缝合肾盂及肾下盏,再用2-0肠线间断全层缝合肾实质切口数针,包膜层用4-0肠线缝合。结果34例均一次性取净结石。术中平均出血量约90ml。术后无大出血病例。结论肾下极背侧肾实质与肾盂联合切开取石术具有术中出血少,无须阻断肾蒂,肾功能受损轻,便于一次取尽结石等优点。适合于肾窦内肾盂特别是肾下盏盏颈相对狭窄的复杂性鹿角形肾结石的治疗。     

Investigation of 89 candidate gene variants for effects on all-cause mortality following acute coronary syndrome

Background  

Many candidate genes have been reported to be risk factors for acute coronary syndrome (ACS), but their impact on clinical prognosis following ACS is unknown.     

甲型血友病基因诊断及产前诊断的研究进展

甲型血友病是最常见的遗传性出血性疾病,受到了国内外研究者们的极大关注.由于此病尚无彻底根治方法,所以基因诊断及产前诊断显得尤为重要.基因诊断可分为直接基因诊断及间接连锁分析的方法;产前诊断的手段也在近年飞速发展.本文就直接基因诊断、间接连锁分析及产前诊断新技术这几方面的研究进展综述如下.     

Hypersensitivity to Forcipomyia taiwana (biting midge): clinical analysis and identification of major For t 1, For t 2 and For t 3 allergens

BACKGROUND: Forcipomyia taiwana is a tiny, blood-sucking midge that cause intense pruritis and swelling in sensitive individuals. It is distributed island-wide in rural Taiwan and Southern China. Objective: This study aimed to study the allergic immune responses and identify F. taiwana allergens. METHODS: Crude whole body F. taiwana extracts were prepared with phosphate-buffered saline. The specific IgE antibody was determined by enzyme-linked immunoassay and immunoblotting. Protein was analyzed by electrospray ionization tandem mass spectrometry. RESULTS: Among the 372 subjects that were exposed to F. taiwana bites, 179 (48%) reported an immediate skin reaction with/without delay reaction and 41(11.1%) reported a solely delay reaction. The skin of 21 subjects was tested with F. taiwana extract. Of these 21 subjects, 12 (57.1%) produced immediate skin reactions and contained high levels of specific IgE antibody against F. taiwana. Immunoblotting revealed that 11 allergenic components are able to bind specific IgE. Allergens of 22, 24, 35, 36, and 64 kDa bound 50, 50, 75, 66.7, and 75% of IgE-containing sera tested, respectively. Tryptic fragments of the 24, 35, 36, and 64 kDa allergens were analyzed by ESI-MS/MS. Selected tryptic peptides of 24, 35, and 36, and 64 kDa allergens exhibited significant sequence identity with triosephosphate isomerase of Anopheles merus,Tenebrio molitor,Ochlerotatus togoi, and Chrysops vittatus, fructose 1,6-bisphosphate aldolase of Antheraea yamamai and Homalodisca coagulata, and a slow muscle myosin S1 heavy chain of Homarusamericanus and a protein with unknown function from A. gambiae, respectively. The 35 and 36 kDa proteins may represent different isoforms of the fructose 1,6-bisphosphate aldolase. CONCLUSION: We conclude that immediate reaction to F. taiwana bites is IgE mediated and the 24 (For t 1), 35 (For t 2), and 64 kDa (For t 3) proteins are candidates for major F. taiwana allergens. Further studies are needed to confirm these allergens.     

抗癌药用植物猫人参(Actinidia macrosperma)离体再生系统的建立

中药猫人参,又名大籽猕猴桃,是著名抗癌药用植物,其野生资源已近枯竭。本文选用猫人参的茎段(至少带一个节)、叶片作为外植体,筛选出了适合猫人参从启动、快繁直至生根等各个阶段的最适培养基,在实验室建立了程序简单、重复性好的再生系统,实现了猫人参组培苗的快速繁殖。以茎段作为外植体效果较好,既容易消毒,同时又能很快诱导腋芽萌发;以叶片作为外植体,能成功诱导出愈伤组织,但此愈伤组织的分化能力极低。以茎段作为外植体获得的无菌培养物在不同发育阶段适宜的培养基组成:外植体启动培养基为MS+BA 5 μmol/L+NAA 1 μmol/L;最适增殖培养基为MS+BA2 μmol/L+GA 1.5 μmol/L+ZT 4μmol/L,35 d繁殖系数为9-10;最适生根培养基为1/2MS+IBA 2μmol/L,生根率达1 00％,培养35 d,平均苗高6.28cm,平均移栽成活率为91％。     

Atrial sympathetic and parasympathetic nerve sprouting and hyperinnervation induced by subthreshold electrical stimulation of the left stellate ganglion in normal dogs

BACKGROUND: Subthreshold electrical stimulation of the left stellate ganglion (LSG) can induce nerve sprouting and sympathetic hyperinnervation in canine ventricles. It is unclear whether a similar neural plasticity involving both sympathetic and parasympathetic innervation also exists in the atria. METHODS AND RESULTS: We applied subthreshold electrical stimulation at 20 Hz (0.45 ms pulse width) or 5 Hz (1.9 ms pulse width) to the LSG in 6 normal mongrel dogs. After 41+/-9 days, the hearts were harvested and the right and left atrium stained for synaptophysin (SYN), growth-associated protein 43 (GAP43), sympathetic nerve markers tyrosine hydroxylase (TH), and parasympathetic marker choline acetyltransferase (ChAT). Tissues from 6 additional healthy dogs were used as controls. The hearts from dogs with LSG electrical stimulation had a higher density of nerve structures immunopositive to the SYN, GAP43, TH, and ChAT (P<.01) in both right and left atria. Nerve density was equal in right and left atria. There were more TH-positive nerve structures than ChAT-positive nerve structures (P<.01) for both right and left atria. No atrial arrhythmia was observed at the second surgery. CONCLUSIONS: Continuous subthreshold electrical stimulation to the LSG induces both sympathetic and parasympathetic hyperinnervation in both right and left atria in normal dogs.     

抗SARS-CoV抗原的人源Fab段噬菌体抗体库的构建

目的 :利用抗SARS冠状病毒IgG抗体阳性的SARS康复患者外周血淋巴细胞 ,构建人源Fab段抗体文库。方法 :制备外周血淋巴细胞总RNA ,逆转录成cDNA。以其为模板 ,利用针对家族特异性Ig基因的引物扩增重链Fd段和轻链基因 ,并重组到噬菌粒载体pComb3中 ,将重组噬菌粒载体电转化大肠杆菌XL 1Blue,酶切鉴定抗体库的重组率 ,并测定噬菌体抗体库的库容量。结果 :构建了源于SARS康复患者血清中抗Fab段的抗体文库 ,轻链、重链Fd段基因的重组率分别为91%和 75 % ,库容量为 7.2 3× 10 7。结论 :成功地构建了抗SARS CoV抗原的人源Fab段噬菌体抗体库