Significance of cytogenetic and fluorescence in situ hybridization analysis in evaluating antichronic myeloid leukemia efficacy of different immune effector cells

We report on the antileukemia effect of interleukin 2 (IL2) on different immune cells from 22 patients with chronic myeloid leukemia (CML). Bone marrow cells from these patients were first cultured in modified long-term bone marrow culture medium for several days, then separately cultured with lymphokine activated killer cells (LAK), cytokine-induced killer cells (CIK), and dendritic cell cocultured CIK (DC-CIK) for another 1-2 days. They were then detected for presence of the Philadelphia chromosome (Ph) by cytogenetic analysis and fluorescence in situ hybridization (FISH). The percentage of Ph-chromosome-positive cells in the bone marrow mononuclear cells after culturing with CIK and DC-CIK was significantly lower than that after culturing with IL2 or LAK. Our results demonstrate that cytogenetics and FISH are useful techniques for the evaluation of the anti-CML effect of immune cells and that CIK or DC-CIK can be appropriate candidates for adoptive immune cell therapy in vivo or for leukemia cell purging ex vivo.     

2型糖尿病患者垂体-肾上腺轴的变化与胰岛素抵抗

目的探讨2型糖尿病患者早期垂体-肾上腺轴的改变与胰岛素抵抗(IR)的关系。方法采用化学发光等方法测定67例新诊断的2型糖尿病患者和33名志愿者(对照组)的血皮质醇(F)和促肾上腺皮质激素(ACTH)(8Am、4Pm和地米抑制后次8Am)以及血清胰岛素(INS)、C肽(C-P)和血糖水平;按HOMA模型计算胰岛素抵抗指数HOM-IR=FINS×FBG/22.5,以此反映2型糖尿病患者IR的情况。结果①2型糖尿病患者8Am血浆F和ACTH、4Pm和地米抑制后次8Am血浆F、血糖、INS、C-P水平以及HOMA-IR均明显高于对照组(P<0.01);②4Pm和地米抑制后次8Am血浆ACTH水平显著低于对照组;③直线相关统计分析显示,皮质醇浓度的升高与空腹血糖升高呈正相关(r=0.48,P<0.01)。与HOMA-IR的升高无直线相关性。结论2型糖尿病患者早期呈现ACTH、皮质醇过度分泌和慢性垂体-肾上腺轴功能紊乱,诱发并加重了糖代谢紊乱和胰岛素抵抗。     

血管内皮损伤标志物在肾移植患者中的应用价值

目的 :通过对肾移植患者血浆可溶性血栓调节蛋白 (solublethrombomdulin ,sTM)、可溶性血管内皮细胞蛋白C受体 (endothelialproteinCreceptor ,sEPCR)、血管性血友病因子 (vonWillebrandfactor,vWF)含量水平的动态检测分析 ,探讨其在肾移植及移植排斥反应中的应用价值。方法 :采用双抗体夹心ELISA法检测 4 2例肾移植患者手术前后、排斥反应治疗过程中的sTM、sEPCR、vWF含量。结果 :肾移植术后sTM、sEPCR、vWF含量水平较术前明显增高 (P <0 0 5 )、与正常对照组比较有显著性差异 (P <0 0 1) ;发生移植排斥反应者较无排斥者有明显差异 (P <0 0 5 ) ;肾移植术前、排斥反应有效治疗后与正常对照组比较无显著差异 (P >0 0 5 ) ;肾移植患者sTM、sEPCR、vWF各指标间呈正相关 (r =0 5 95 ,P <0 0 1,r =0 5 81,P <0 0 1,r =0 6 0 6 ,P <0 0 1)。结论 :sTM、sEPCR、vWF均可作为肾移植患者血管内皮的免疫损伤标志物 ,联合动态监测sTM、sEPCR、vWF的含量在观察肾移植手术创伤程度、排斥反应的早期诊断和治疗效果的监测等方面均有重要的临床应用价值。     

骨组织工程用聚磷酸钙的结构和性能的研究

多孔聚磷酸钙（Calcium polyphosphate，CPP）生物陶瓷是一种新型的骨组织工程支架材料，国外有研究表明聚磷酸钙作为骨组织工程支架材料具有良好的生物相容性及可降解性。通过对原料煅烧过程的研究，探索了聚合度的计算方法，制备出不同聚合度的材料。通过对材料烧结温度的试验，制得了不同晶型的聚磷酸钙多孔材料。在Tris—HCl缓冲液中进行的体外降解实验表明，CPP多孔支架材料是可控降解的，不同聚合度和晶型的支架材料力学强度和降解性能不同。随着聚合度的增加，材料的力学强度增大，降解速率变小；非晶态的CPP17d就可以完全降解，7晶型的CPP25d可完全降解，β和α晶型的CPP降解缓慢，30d分别降解了大约12％和5％。因此聚磷酸钙是一种很有前途的骨组织工程支架材料，凭借其独特的无机聚合物结构及降解性能，有望实现可控速率的降解。     

Timosaponin B-II improves memory and learning dysfunction induced by cerebral ischemia in rats

Sapogenins from Anemarrhenae asphodeloides was reported to improve the learning and memory abilities. In this study, we investigated the effect of Timosaponin B-II(TB-II), a purified extract from A. asphodeloidesb on rat vascular dementia (VD) produced by transient (2 h) middle cerebral artery occlusion. The learning and memory abilities of rats were measured by water maze task and passive avoidance task. Daily oral administration of TB-II at two different dose levels of 100 and 200 mg/kg resulted in a significant improvement of the deficit in the learning of the water maze task, beginning 14 days after ischemia. Shortened mean escape latency was detected in TB-II group compared with model group during the same trial days. TB-II treatment also significantly reversed the ischemia-induced retention deficit determined by a one trial step-down type of passive avoidance task. Meanwhile, the expression of interleukin-10, an anti-inflammatory cytokine, and its receptor were significantly increased in TB-II treated VD rats. The results presented the first evidence of a neuroprotective effect of TB-II in the model of vascular dementia. We suggest that the anti-dementia effect by TB-II is derived at least in part from its anti-inflammatory properties.     

Population survey of CCR5 delta32, CCR5 m303, CCR2b 64I,and SDF1 3'A allele frequencies in indigenous Chinese healthy individuals,and in HIV-1-infected and HIV-1-uninfected individuals in HIV-1 risk groups

The aim of this study is to determine in indigenous Chinese ethnic groups the frequencies of the chemokine (SDF1 3'A) and chemokine receptors (CCR5 delta32, CCR5 m303, and CCR2b 64I) HIV-1/AIDS restriction alleles. The study includes two cohorts; the first comprised 3165 indigenous healthy subjects representing eight ethnic groups: Han (n = 1406), Uygur (n = 316), Mongolia (n = 134), Hui (n = 386), Tibetan (n = 330), Zhuang (n = 378), Dai (n = 101), and Jingbo (n =114). The second cohort consisted of 330 HIV-1-infected (86 subjects infected by sexual transmission and 198 subjects infected by HIV-1-contaminated blood or by sharing injection equipment; the remaining 46 subjects said nothing about HIV-1 transmission) and 474 HIV-1-uninfected Han Chinese belonging to one of two HIV-1 high-risk groups: intravenous drug users (n = 215) and individuals with sexually transmitted diseases (n = 259). Genotypes for the four genes were obtained using PCR (CCR5 delta32) or PCR-restriction fragment length polymorphism. Randomly selected amplified PCR products were further confirmed by direct DNA sequencing. The variant allele frequencies were determined to be 0% to 3.48% for CCR5 delta32, 0% for CCR5 m303, 16.23% to 28.79% for CCR2b 64I, and 17.70% to 27.76% for SDF1 3'A in Chinese healthy individuals from eight ethnic groups. These findings show that allele frequencies differ among the eight Chinese ethnic groups for CCR5 delta32, CCR2b 64I, and SDF1 3'A and that the CCR5 m303 and CCR5 delta32 mutant alleles were absent or infrequent in Chinese, which may be helpful for studies of specific anti-HIV-1 vaccine trials and coreceptor inhibitor drug targets in Chinese populations. Furthermore, we observed no significant differences in allele or genotypic frequencies between HIV-1-infected and HIV-1-uninfected groups from the Han ethnic group. Our finding is the first reporting that there is likely no effect of the examined polymorphisms in our study on HIV-1 transmission in the Chinese Han population, However, the genetic effects of these and other AIDS-modifying polymorphisms on the pathogenesis and clinical outcome of HIV-1/AIDS diseases is under investigation in Chinese populations.     

Organ distribution of severe acute respiratory syndrome (SARS) associated coronavirus (SARS-CoV) in SARS patients: implications for pathogenesis and virus transmission pathways

We previously identified the major pathological changes in the respiratory and immune systems of patients who died of severe acute respiratory syndrome (SARS) but gained little information on the organ distribution of SARS-associated coronavirus (SARS-CoV). In the present study, we used a murine monoclonal antibody specific for SARS-CoV nucleoprotein, and probes specific for a SARS-CoV RNA polymerase gene fragment, for immunohistochemistry and in situ hybridization, respectively, to detect SARS-CoV systematically in tissues from patients who died of SARS. SARS-CoV was found in lung, trachea/bronchus, stomach, small intestine, distal convoluted renal tubule, sweat gland, parathyroid, pituitary, pancreas, adrenal gland, liver and cerebrum, but was not detected in oesophagus, spleen, lymph node, bone marrow, heart, aorta, cerebellum, thyroid, testis, ovary, uterus or muscle. These results suggest that, in addition to the respiratory system, the gastrointestinal tract and other organs with detectable SARS-CoV may also be targets of SARS-CoV infection. The pathological changes in these organs may be caused directly by the cytopathic effect mediated by local replication of the SARS-CoV; or indirectly as a result of systemic responses to respiratory failure or the harmful immune response induced by viral infection. In addition to viral spread through a respiratory route, SARS-CoV in the intestinal tract, kidney and sweat glands may be excreted via faeces, urine and sweat, thereby leading to virus transmission. This study provides important information for understanding the pathogenesis of SARS-CoV infection and sheds light on possible virus transmission pathways. This data will be useful for designing new strategies for prevention and treatment of SARS.     

Inhibitory effect of intrathecal meptazinol on carrageenan-induced thermal hyperalgesia in rats

The effect of meptazinol in the spinal cord on carrageenan-induced hyperalgesia was investigated. The latency of paw withdrawal (PWL) to a thermal stimulus was used as an index of inflammatory hyperalgesia in awake rats. Intrathecal (i.t.) injection of 10 and 100 microg meptazinol markedly increased the PWL of the carrageenan-injected paw (P<0.01). The PWL of the non-injected paw was not detectably affected by the administration of meptazinol at the doses tested. I.t. injection of naloxone (5 microg) or atropine (1 microg) alone exhibited no effect on the PWLs of either the carrageenan-injected or non-injected paw. Pretreatment with naloxone, but not atropine, completely blocked the meptazinol-induced anti-hyperalgesia. These observations suggested that mu opioid receptor rather than muscarinic acetylcholine receptor may be involved in the anti-hyperalgesia of meptazinol in the spinal cord.