AIM To examine hepatitis C in hepatocellular carcinoma in most endemic area, Guangxi, China.METHODS Immunochemistry was performed on formalin fixed, paraffin embedded tissue sections. A total202 specimens were analysed from the high, intermediate and low HCC prevalent regions of Guangxi.RESULTS The distribution of positive hepatitis C cases from high, intermediate and low regions wasrespectively 24/63 (38%), 23/62 (37%) and 30/77 (39%), with a total of 77/202 (38.12%).CONCLUSION Hepatitis C virus is an important risk factor in the development of hepatocellularcarcinoma, but the regional difference in prevalence of this cancer is more likely influenced by hepatitis Bviral infection and aflatoxin B1 exposure. In Guangxi, infection of hepatitis B and C virus in thedevelopment of hepatocellular carcinoma may be greatly enhanced by exposure to aflatoxin.

mycobacterium avium; paratuberculosis; glycosyltransferases; bacillus subtilis;chromatography, affinity; antibodies, anti-idiotypic

免疫毒素2E8-去甲斑蝥素的研制及其体外靶向抑制效应研究

Objective The immunotoxins generated by conjugating monoclonal antibody (mAb) and a certain toxin play an important and promising role in treating hematopoietic malignancies. However, most of the toxins used for the conjugation are toxic proteins, which are immunogenic in the patients. Norcantharidin (NCTD) is a small molecule toxin without immunogenicity, and thus has become a potential new drug for hematopoietic cancers. In this study, we prepared immunotoxin 2E8-NCTD by using the ZCH-4-2E8 cells produced in the laboratory of our hospital, and then detected its targeting effect against CD+19 lymphoid malignant Nalm-6 cells in vitro.Methods 2E8 mAb was obtained from mouse ascites and purified by gel chromatography. After its purity was checked by SDS-PAGE, immunotoxin 2E8-NCTD was generated by conjugating CD19 mAb with NCTD using activated ester method. The binding activity of the immunoconjugate to CD19 antigens on cell surface, and the expression levels of the CD19 antigens on Nalm-6 and K562 cells were determined by flow cytometry. The inhibitory effects of PBS, purified 2E8 mAb, NCTD, and immunotoxin 2E8-NCTD on the cell growth of either Nalm-6 or K562 cells were then compared.Results The purity of the 2E8 mAb was higher than 99% demonstrated by SDS-PAGE assay. 2E8 mAb was detected on the surface of 99.34% of the Nalm-6 cells, while on only 0.98% of the K562. The newly generated immunotoxin had a positive rate of 99.90% on the Nalm-6 with slightly reduced binding activity. Both 2E8-NCTD and NCTD significantly inhibited the growth of CD+19 Nalm-6 cells (P < 0. 001 ), while the purified 2E8 mAb did not show any significant influences on the growth of the same cell line ( P > 0.05 ). Meanwhile, no significant inhibitory effects on the CD-19 K562 cells were identified in the 2E8-NCTD, 2E8 mAb, or control groups, indicating a significant targeting effect of 2E8-NCTD against Nalm-6 cells.Conclusions The immunotoxin 2E8-NCTD can be synthesized by activated ester method. It has target killing effects on CD+19 Nalm-6 leukemia cells in vitro.

Since the approval of rituximab in 1997, monoclonal antibodies (mAbs) have become an increasingly important component of therapeutic regimens in oncology. hTe success of mAbs as a therapeutic class is a result of great strides that have been made in molecular biology and in biotechnology over the past several decades. Currently, there are 14 approved mAb products for oncology indications, and there are ten additional mAbs in late stages of clinical trials. Compared to traditional chemotherapeutic agents, mAbs have several advantages, including a long circulating half-life and high target speciifcity. Antibodies can serve as cytotoxic agents when administered alone, exerting a pharmacologic effect through several mechanisms involving the antigen binding (Fab) and/or Fc domains of the molecule, and mAbs may also be utilized as drug carriers, targeting a toxic payload to cancer cells. hTe extremely high affnity of mAbs for their targets, which is desirable with respect to pharmacodynamics (i.e., contributing to the high therapeutic selectivity of mAb), otfen leads to complex, non-linear, target-mediated pharmacokinetics. In this report, we summarize the pharmacokinetic and pharmacodynamics of mAbs that have been approved and of mAbs that are nearing approval for oncology indications, with particular focus on the molecular and cellular mechanisms responsible for their disposition and effcacy.

Objective:To study the metabolism abnormality of homocysteine(Hcy) in unexplained repeated spontaneous abortion(URSA).Methods:The level of Hcy in sera was measured with hyperpressure liquid chromatography(HPLC);Folic acid and vitamin B12 were detected by radioimmune assay;anticardiolipin antibody (ACA) was detected by ELISA.Results:(1)The level of serum Hcy in URSA group was significantly higher than that in control group,showing a statistical significant difference(P＜0.01).The level of Hcy was correlated with ages,but not correlated with areas,numbers of miscarriage,gestation age,primary or secondary abortions.(2)The levels of folic acid and vitamin B12 in URSA group were significantly lower than those in control group.The levels of serum folic acid and vitamin B12 were not correlated with age,area,numbers of miscarriage and abortion periods.(3)ACA positive rate in URSA was significantly higher than that in control group.The level of Hcy in ACA(+) group was significantly higher than that in ACA(-) group among URSA patients.Conclusions:Hyperhomocysteinemia,low folic acid state,and ACA were all the independent risk factors for URSA.Lacking of folic acid and vitamin B12 is one of the important causes of hyperhomocysteinemia.ACA and hyperhomocysteinemia may have synergistic action in the occurrence of URSA.

Objective The study will explore effects of the autoantibodies against AT1 receptor and angiotensin Ⅱ on the refractory hypertension. Methods Seventy-seven patients (46 men and 31 women) with essential hypertension were divided into groups of refractory hypertension (RH) and hypertension (HT) according to the 1999 WHO -ISH Guidelines for the Management of Hypertension. Forty normotensives (22 men) were recruited as controls.The mean age was 54. 3 ± 13 years old in RH group,53.5±9 years old in HT group and 51.2±11.9years old in normotensives (NT) group. The mean blood pressure was 154.2 ± 9.4/98.4 ± 8.2 mmHg in RH group and 130.1 ±7.6/80.5 ±6.7 mmHg in HT group after combination drug therapy of hypertension for 4 weeks. Blood pressure in NT group was 120. 8 ± 11.7/76. 4 ± 7.2 mmHg. The epitope of the 2nd extracellular loops of AT1 receptor was synthesized and used as antigens to screen the autoantibodies by ELISA. Plasma angiotensin (Ang) Ⅱ were examined by a radioimmunoassay. Results The autoantibodies against AT1 receptor were positive in 18 (46. 15% ) patients with RH, in 4 (10. 5 % ) hypertension and in 3 (7.5 % ) normotensives, P ＜ 0.01. Ang Ⅱwas 57.01 ± 52.63 pmol/L in patients with RH. Both the autoantibodies positive and the Ang Ⅱ increasing were 4 (10. 3 %) cases, both normal were 7 (17.9% ) cases, the autoantibodies positive or Ang Ⅱ in creasing was all of 14 (35.9 % ) cases (χ2 =0. 09,P ＞ 0. 05) There was no relationship between the autoantibodies against AT1 receptor and the angiotensin Ⅱ in refractory hypertension. Conclusion The autoantibodies against AT1 receptor and Ang Ⅱ might be two independent factors in developing of refractory hypertension. The findings suggest that AT1 receptor antagnist used in the treatment of refractory hypertension might have an important value.