Molecular cloning of cDNA coding for the 68 kDa allergen of Penicillium notatum using MoAbs

To characterize the 68 kDa allergen of Penicillium notatum (also known as P. chrysogenum), a molecular antibody (MoAb) (P40) was previously generated. For cDNA cloning, three more MoAbs (3F, 5A3, 5G2) were generated in the present study. A mixture of all the four MoAbs was used in cloning of the gene coding for the 68 kDa allergen from a λgt11 cDNA library of P. chrysogenum. A cDNA clone (A6) with DNA insert of about 0.5 kb which encodes for the 3′-terminal nucleotide sequence of the 68 kDa allergen was obtained. The cloned sequence contained two putative N-glycosylation sites. The reduction in molecular weight from 68 to 62 kDa in immunoblotting after treatment of the crude extract of P. notatum with N-glycosidase F indicates that the 68 kDa allergen is a glycoprotein. Nucleotide sequence determination showed that 188 (54%) of the 348 nucleotides of the cDNA sequence obtained were identical to the same region of the nucleotide sequence of the beta-N-acetylglucosaminidase gene of Candida albicans. Although the cDNA clone obtained did not encode the full-length gene of the 68 kDa allergen, polypeptide expressed from the A6 cDNA showed positive immunological reactivities to all four MoAbs used in the cloning experiment and to IgE antibodies in sera of asthmatic patients. There was a loss of immunoblotting activity to the 68 kDa component after absorption of MoAb P40-containing culture supernatant with filters blotted on plaque lawns of cDNA clone A6. Moreover, the immunoblotting activity remained when the MoAbs affinity-purified with filters containing polypeptides encoded by the cDNA insert of clone A6 were used. These two observations indicate that clone A6, which encodes 117 amino acid residues of a putative 560-residue polypeptide, is a cDNA clone of the 68 kDa component of P. notatum. In conclusion, results obtained from cloning and characterization of a partial cDNA clone described in this study suggest that the 68 kDa allergen of P. notatum is a beta-N-acetylglucosaminidase.     

Changes of hepatic and systemic haemodynamics following somatostatin administration in patients with hepatitis B-related cirrhosis

Abstract Somatostatin has been used to effectively control acute variceal haemorrhage, with conjectured mechanisms on portal hypertension. We, therefore, evaluated the effects of somatostatin on hepatic and systemic haemodynamics in 15 patients with hepatitis B-related cirrhosis and portal hypertension. All patients received an intravenous, continuous infusion of somatostatin 250 μg/h, following a bolus injection of 250 μg. In systemic haemodynamics, the mean arterial pressure (MAP) increased ( P < 0.05), associated with a reflex bradycardia within 3 min following bolus injections, compared with basal values. The right atrial pressure, pulmonary capillary wedge pressure, inferior vena cava pressure, cardiac index, and systemic vascular resistance remained unaffected after drug infusion. In hepatic haemodynamics, the wedge hepatic vein pressure remained unchanged after drug administration. However, there was an increase in free hepatic vein pressure (FHVP; P < 0.05), and a trend toward a decrease in the hepatic vein pressure gradient (HVPG; P = 0.063), within 3 min after bolus injection. Furthermore, the hepatic blood flow decreased significantly at 10 and 30 min after somatostatin infusion ( P < 0.05). The effective sinusoidal perfusion assessed by indocyanine green infusion also decreased progressively at 10 min ( P = 0.057) and 30 min ( P < 0.05). We concluded that somatostatin, at the dose used in this study, caused a transient and bolus-related vasoconstrictive effect, resulting in increases in MAP and FHVP, a decrease in heart rate, and a trend toward lower HVPG. In addition, somatostatin reduced the hepatic blood flow and effective sinusoidal perfusion which may be hazardous to cirrhotic patients during variceal haemorrhage.     

γ-Pyrone Compounds as Potential Anti-cancer Drugs

Abstract— The γ-pyrones, artomunoxanthotrione epoxide, cyclocommunol, cyclomulberrin, and cyclocommunin exhibited potent inhibition of human PLC/PRF/5 and KB cells in-vitro. Dihydroisocycloartomunin showed significant and potent inhibition of human PLC/PRF/5 and KB cells in-vitro, respectively. Cyclomorusin, dihydrocycloartomunin and artomunoxanthone showed significant inhibition of KB cells in-vitro. Based on the above finding and the reported antileukaemic activity of xanthone psorospermin, a series of natural γ-pyrones was prepared and the inhibition of human PLC/PRF/5 and KB cells in-vitro was measured. Structure-activity analysis indicated the epoxide group substituted at 3-hydroxyl and 2,6-; 3,6-; and 3,5-dihydroxyl xanthone enhanced the anti-tumour activity. The epoxide group substituted at the 6-hydroxyl group of 1,6-dihydroxyxanthone did not show anti-tumour activity.     

环孢素A体外诱导HL—60细胞凋亡的研究

目的：观察环孢素Ａ是否有诱导白血病细胞凋亡的作用。方法：应用细胞形态学检查、二苯胺法ＤＮＡ片段化定量，ＤＮＡ凝胶电泳及流式细胞术等方法观察细胞凋亡。结果：ＣｓＡ５０ｍｇ／Ｌ作用ＨＬ－６０细胞４ｈ，ＤＮＡ片段率显著增高，达（２８．２±５．８）％，而对照组仅为（１２．５±１．７）％（Ｐ＜０．０１，ｎ＝１０）。光镜及电镜检查见细胞核固缩，碎裂，有凋亡小体形成。ＤＮＡ电泳显示典型的ＤＮＡｌａｄｄｅｒ。流式细胞术检测发现ＣｓＡ５０ｍｇ／Ｌ作用ＨＬ－６０细胞６ｈ凋亡细胞率为４９．７％，而空白组仅为９．１％。ＣｓＡ诱导ＨＬ－６０细胞ＤＮＡ片段化呈剂量和时间依赖性。结论：ＣｓＡ在体外能诱导ＨＬ－６０细胞凋亡。     

CD3ζ链基因在脐带血T细胞及其CD4+和CD8+T细胞亚群中的表达特点

目的 了解脐带血T细胞及其CD4 和CD8 T细胞亚群的信号传导分子CD3ζ基因的表达特点.方法 采用SYBR Green Ⅰ荧光定量PCR和相对定量分析法检测60例脐带血单个核细胞和12例纯化脐带血CD4 及CD8 T细胞的CD3ζ基因的表达情况,以β2微球蛋白基因(β2M)作为内参,采用相对定量公式:2-△Ct×100%,计算CD3ζ基因相对mRNA表达量,60例健康成人作为对照.并根据荧光定量PCR熔解曲线特点和序列分析了解CD3ζ基因突变情况.结果 全部脐带血和健康成人外周血单个核细胞均表达CD3ζ基因,不同个体CD3ζ基因表达量有所差异,脐带血T细胞CD3ζ基因相对mRNA表达量为6.7%±5.56%,CD4 和CD8 T细胞的CD3ζ基因相对mRNA表达量分别为6.74%±2.0%和6.88%±1.76%,三者CD3ζ基因表达量均明显高于健康成人(P=0.000,P=0.034,P=0.000).序列分析结果显示尚未发现国外文献所报道的ζ链剪接异构体.结论 本研究率先报道了脐带血T细胞及其CD4 和CD8 T细胞亚群的CD3ζ基因mRNA的表达水平,为进一步了解脐带血T细胞免疫学特点提供新的基础资料.     

神经营养素受体在人外周血T细胞中表达的研究

目的 研究人外周血CD4^ /CD8^ T细胞4种神经营养素受体基因的转录。方法 应用尼龙手法分离出T细胞，磁式细胞分离法（MACS）分离CD4^ /CD8^ T细胞亚群，再以RT－PCR法研究4种神经营养素受体在两种T细胞亚群上的表达。结果 未经刺激的CD4^ /CD8^ T细胞亚群不表达任何神经营养素受体。经PHA或PPD刺激后，CD4^ /CD8^ T细胞亚群表达trkA,CD8^ T细胞亚群表达trkC；而在各种状态下的T细胞上均未见表达trkB及p75^NGFR。结论神经营养素受体在两种T细胞亚群中有不同的表达格局，提示不同T细胞亚群受神经营养素调节的模式可能各不相同。     

Four-Dimensional Computerized Tomography （4D-CT） Reconstruction Based on the Similarity Measure of Spatial Adjacent Images

Objective：To investigate the feasibility of a 4D-CT reconstruction method based on the similarity principle of spatial adjacent images and mutual information measure. Methods：A motor driven sinusoidal motion platform made in house was used to create one-dimensional periodical motion that was along the longitudinal axis of the CT couch. The amplitude of sinusoidal motion was set to an amplitude of ±1 cm. The period of the motion was adjustable and set to 3.5 s. Phantom objects of two eggs were placed in a Styrofoam block, which in turn were placed on the motion platform. These objects were used to simulate volumes of interest Undergoing ideal periodic motion. CT data of static phantom were acquired using a multi-slice general electric （GE） LightSpeed 16-slice CT scanner in an axial mode. And the CT data of periodical motion phantom were acquired in an axial and cine-mode scan. A software program was developed by using VC ＋ ＋ and VTK software tools to resort the CT data and reconstruct the 4D-CT. Then all of the CT data with same phase were sorted by the program into the same series based on the similarity principle of spatial adjacent images and mutual information measure among them, and 3D reconstruction of different phase CT data were completed by using the software. Results：All of the CT data were sorted accurately into different series based on the similarity principle of spatial adjacent images and mutual information measures among them. Compared with the unsorted CT data, the motion artifacts in the 3 D reconstruction of sorted CT data were re- duced significantly, and all of the sorted CT series result in a 4D-CT that reflected the characteristic of the periodical motion phantom. Conclusion：Time-resolved 4D-CT reconstruction can be implemented with any general multi-slice CT scanners based on the similarity principle of spatial adjacent images and mutual information measure. The process of the 4D-CT data acquisition and reconstruction were not restricted to the hardware or software of the C     

rhGM-CSF作为成人乙肝疫苗无(弱)应答者免疫佐剂初探

Objective To investigate the effect of recombinant human granulocyte macrophage colony stimulating factor (rhGM-CSF) as adjuvant on immune response in adults of non-and hyporesponders to hepatitis B vaccine. Methods Those who were once immunized with recombined yeast gene hepatitis B vaccine more than one standard scheme in two years and negative for hepatitis B markers were randomly sorted as group A and group B. 33 adults of group A were given hepatitis B vaccine 10 μg each time. The immune procedure was O, 1 and 6month. 34 adults of group B were given rhGM-CSF 300 μg for the first day, then 10 μg each time for routine immune. The blood samples were collected before the first injection and in 1, 2 and 8 months (T1, T2, TS)following the first injection to test Anti-HBs. Results Anti-HBs positive conversion rates of group A and B at T8 was 39.39% and 64.71% respectively(P=0.038). Anti-HBs levels of group B at TI, T2, T8 were(113.85±198.56) mIU/ml, (312.40±349.44) mIU/ml, (427.74±411. 58) mIU/ml (P=0.001). There was significant difference between group A and B in T8 Anti-HBs levels(P=0.010). Conclusion Better immune response was found in the group of rhGM-CSF with hepatitis B vaccine. So rhGM-CSF can induce the immune respond to hepatitis B vaccine.     

A COMPARISON OF ATRACURIUM AND ALCURONIUM DURING HALOTHANE ANAESTHESIA BY MEASUREMENT OF THE TRAIN-OF-FOUR RESPONSE OF THE ADDUCTOR POLLICIS MUSCLE AND CLINICAL OBSERVATION

Atracurium and alcuronium have been compared during halothaneanaesthesia, by measurement of the mechanical response of theadductor pollicis muscles to train-of-four stimulation and byclinical observation. Atracurium appeared significantly shorter-actingthan alcuronium. However, results suggested that the actionof alcuronium may not be of "medium duration ". A comparisonof three indices of muscle twitch response to the train-of-fournerve stimulation, seemed to indicate that the D'/D ratio gavethe best overall index of neuromuscular blockade in this study.