中国人CCR5732基因突变的检测及其对HIV遗传易感性的初步评估

为了解ＣＣＲ５Δ３２ 基因突变在中国本土人群基因组中的分布,初步评估我国人群对ＨＩＶ－１ 感染的遗传易感性,用ＰＣＲ扩增、Ｓｏｕｔｈｅｒｎ 杂交和ＤＮＡ 测序等分子生物学方法对９１５ 名中国人来源的基因组ＤＮＡ中ＣＣＲ５Δ３２ 基因突变进行了检测。结果发现,所有被检测的个体绝大多数均表现为ＣＣＲ５ ｗ ｔ／ｗ ｔ纯合子等位基因型,仅检测到两例个体为突变的杂合子ＣＣＲ５ｗ ｔ／Δ３２ 基因型,而未见到有突变的ＣＣＲ５Δ３２／Δ３２ 纯合子的个体。上述初步结果提示：在我国人群中,ＣＣＲ５Δ３２ 基因突变率很低（约为０．２％ ）,所以我国绝大多数人群对ＮＳＩ嗜巨噬细胞的ＨＩＶ－１ 株感染的遗传易感性较高     

中国人CCR5Δ32基因突变的检测及其对HIV遗传易感性的初步评估

为了解ＣＣＲ５Δ３２基因突变在中国本土人群基因组中的分布,初步评估我国人群对ＨＩＶ－１感染的遗传易感性,用ＰＣＲ扩增,Ｓｏｕｔｈｅｒｎ杂交和ＤＮＡ测序等分子生物学方法对９１５名中国人来源的基因组ＤＮＡ中ＣＣＲ５Δ３２基因突变进行了检测。结果发现,所有被检测的个体绝大多数均表现为ＣＣＲ５ｗｔ／ｗｔ纯合子等位基因型,仅检测到两例个体为突变的杂合子ＣＣＲ５ｗｔ／Δ３２基因型,而未见到有突变的ＣＣＲ５Δ３     

北京地区发生庚型肝炎病毒感染的初步研究

庚型肝炎病毒（ＧＢＶ－Ｃ／ＨＧＶ）是最近被证实的一种致人类肝炎病毒，用ＲＴ－ＰＣＲ法以ＧＢＶ－Ｃ的５′端非编码区的序列为引物，对北京地区４１例非Ａ～Ｅ病毒性肝炎患者检测，发现２６．８％的ＧＢＶ－Ｃ感染，他们的临床表现为，ＡＬＴ反复低水平异常，具有轻型慢性肝炎的表观，值得注意的是，虽然ＧＢＶ－Ｃ可通过血液传播但本组大部分ＧＢＶ－Ｃ患者无输血史，说地区较高ＧＢＶ－Ｃ感染率值得重视。     

PCT检测HCV-RNA阳性与血清HCV抗体的对比研究

ＰＣＴ检测ＨＣＶ－ＲＮＡ阳性与血清ＨＣＶ抗体的对比研究武警河南总队医院病理科梁喜林，郑晓芙，彭亚丁，粟娟（郑州４５００５２）关键词ＰＣＲ检测，ＨＣＶ－ＲＮＡ，抗－ＨＣＶ采用ＰＣＲ技术检测ＨＣＶ－ＲＮＡ及酶免检测血清丙型肝炎抗体（抗－ＨＣＶ）来测定及辅...     

脑创伤后脑内IL-1β、细胞间粘附分子-1、血管细胞粘附分子-1的变化

目的探讨脑创伤后脑内白细胞介素－１β（ＩＬ－１β）、细胞间粘附分子－１（ＩＣＡＭ－１）及血管细胞粘附分子－１（ＶＣＡＭ－１）与继发性脑损伤（ＳＢＩ）发病的关系。方法用原位杂交（ＩＳＨ）、免疫细胞化学（ＩＣＣ）等方法，观察了脑创伤后脑内ＩＬ－１β、ＩＣＡＭ－１及ＶＣＡＭ－１的变化。结果侧脑室注射重组人ＩＬ－１β（ｒｈＩＬ－１β），显著增加多形核粒细胞（ＰＭＮ）在受伤脑组织中的侵入和聚集；而给予ＩＬ－１β抗体和Ｎ－乙酰半胱氨酸（ＮＡＣ），则明显减少ＰＭＮ在受伤脑组织中的侵入和聚集。提示，脑创伤后受伤脑组织、ＩＬ－１β、ＩＣＡＭ－１及ＶＣＡＭ－１的表达分泌增加与ＰＭＮ在受伤脑组织中的侵入和聚集明显相关。结论：脑创伤后脑内炎症反应，包括脑创伤后ＰＭＮ在受伤脑组织内侵入和聚集，可能在ＳＢＩ发病过程中起重要作用。     

与ColE1相容的高效表达载体的构建及用于HIV-1蛋白酶在大肠杆菌的表达

描述了两套与ＣｏｌＥ１相容的Ｅ．ｃｏｌｉ表达载体的构建过程。表达载体的复制起点来源于ｐＡＣＹＣ１７７，用此表达的蛋白可以方便地与通常使用的ＣｏｌＥ１来源的表达载体表达的蛋白共表达。这些载体带有Ｋｍ筛选标记，利用Ｔ７噬菌体的ｇ１０的先导序列增强翻译起始。已用于有活性的重组ＨＩＶ－１蛋白酶在Ｅ．ｃｏｌｉ胞质中的高效表达。重组蛋白酶与天然蛋白酶序列一致，且具有离体与在体的切割活性。从中可以纯化出大量的ＨＩＶ－１蛋白酶，满足生化分析以及抑制剂筛选的需求     

HIV—1多表位膜蛋白在E.coli中的表达及在血清检测中的应用

目的：获得具有较多保守抗原表位的ＨＩＶ－１膜重组抗原，提高ＨＩＶ筛选ＥＬＩＳＡ试剂的质量。方法：利用分子克隆与ＰＣＲ方法，获得了新的ＨＩＶ－１膜重组抗原的基因克隆，编码ｇｐ１２０Ｃ端亲水区３７个氨基酸残基（ｅｎｖ４８２～５１８）以及ｇｐ４１膜外部分的１２８个氨基酸残基（ｅｎｖ５４８～６７５）。将该基因克隆到ｐＧＥＭＥＸ－１载体上融合表达，表达蛋白经纯化后，以１μｇ／ｍｌ的浓度包被ＥＬＩＳＡ板，用来     

人源抗HIV—1整合酶单链抗体基因的构建及表达

获得人源抗ＨＩＶ－１整合酶单链抗体基因并在大肠杆菌及细胞中进行表达。方法：从人的抗ＨＩＶ－１整合酶的Ｆａｂ抗体基因片段中通过ＰＣＲ扩出此抗体的重链可变区和轻链可变区基因,经一弹性连接肽连接构建成人源抗ＨＩＶ－１整合酶的单链抗体基因。     

替尼泊苷为主的联合方案治疗难治性复发性急性白血病的疗效及多药耐药基因的表达

用替尼泊苷（威猛，ＶＭ－２６）为主的联合方案治疗了３０例难治性复发性急性白血病并用链亲和素胶体金原位杂交（ＩＳＨ－ＳＡＧ）研究了多药耐药基因（ｍｄｒ－１）表达对疗效的影响。３０例病人中完全缓解１５例（ＣＲ），部分缓解（ＰＲ）５例，ＣＲ率５０．０％，有效率６６．７％。２１例检测了ｍｄｒ－１基因表达，其中ｍｄｒ－１阳性者７１．４％，阴性者２８．６％。ｍｄｒ－１阳性组ＣＲ率２６．７％，阴性组ＣＲ率８３．３％（Ｐ＜０．０５）。１５例ｍｄｒ－１基因阳性病人中，加用环孢素Ａ逆转治疗者的ＣＲ率５０％，未用逆转治疗者的ＣＲ率１２．５％。     

低硒病毒性心肌炎鼠IL—2及SIL—2R的检测

目的：研究硒与ＩＬ－２活性及ＳＩＬ－２Ｒ水平的关系。方法：采用不同含量的硒饲料喂养小鼠,接种ＣＶＢ病毒,建立心肌炎模型后,采用生物活性法检测ＣｏｎＡ刺激下小鼠脾细胞培养上清中ＩＬ－２活性。并采用双抗体夹心法检测其ＳＩＬ－２Ｒ水平。结果：低硒条件下,病毒感染鼠ＩＬ－２Ｒ水平低于常硒病毒组,结论：低硒、ＣＶＢ３Ｍ感染可使小鼠免疫功能紊乱。     

Rational development of radiopharmaceuticals for HIV-1

The global battle against HIV-1 would benefit from a sensitive and specific radiopharmaceutical to localize HIV-infected cells. Ideally, this probe would be able to identify latently infected host cells containing replication competent HIV sequences. Clinical and research applications would include assessment of reservoirs, informing clinical management by facilitating assessment of burden of infection in different compartments, monitoring disease progression and monitoring response to therapy. A “rational” development approach could facilitate efficient identification of an appropriate targeted radiopharmaceutical. Rational development starts with understanding characteristics of the disease that can be effectively targeted and then engineering radiopharmaceuticals to hone in on an appropriate target, which in the case of HIV-1 (HIV) might be an HIV-specific product on or in the host cell, a differentially expressed gene product, an integrated DNA sequence specific enzymatic activity, part of the inflammatory response, or a combination of these. This is different from the current approach that starts with a radiopharmaceutical for a target associated with a disease, mostly from autopsy studies, without a strong rationale for the potential to impact patient care. At present, no targeted therapies are available for HIV latency, although a number of approaches are under study. Here we discuss requirements for a radiopharmaceutical useful in strategies targeting persistently infected cells. The radiopharmaceutical for HIV should be developed based on HIV biology, studied in an animal model and then in humans, and ultimately used in clinical and research settings.     

伴HIV感染的肛管尖锐湿疣激光治疗疗效观察

目的探讨C02激光治疗伴艾滋病毒（Human immunodeficiency virus,HIV）的肛管尖锐湿疣（condyloma acuminatum,CA）的临床疗效。方法回顾性分析2010年1月至2012年6月;肛管CA患者262例,其中18例患者HIV感染。将患者分为两组,肛管CA伴HIV感染者18例,单纯肛管CA感染者244例,均采用CO2激光治疗后,比较两组患者的病情严重程度和临床平均治愈次数。结果肛管CA伴HIV感染组病情程度平均计分2．77分,单纯肛管CA组病情程度平均计分2．47分,两组患者病情程度平均计分比较,差异有显著意义（P〈0．05）。两组患者治疗痊愈所用平均治疗次数,HIV感染组为5．1次,单纯肛管CA组3．0次,差异有显著意义（P〈0．05）。结论肛管CA伴HIV感染者病情严重程度和临床平均治愈次数高于不伴HIV感染者。     

Avian influenza: potential impact on sub-Saharan military populations with high rates of human immunodeficiency virus/acquired immunodeficiency syndrome

Several sub-Saharan militaries have large percentages of troops with human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome. With the arrival of avian influenza in Africa, the potential exists that some of those soldiers might also become infected with H5N1, the virus responsible for the disease. Two possible scenarios have been postulated regarding how such a coinfection of HIV and H5N1 might present. (1) Soldiers already weakened by HIV/acquired immunodeficiency syndrome rapidly succumb to H5N1. The cause of death is a "cytokine storm," essentially a runaway inflammatory response. (2) The weakened immune system prevents the cytokine storm from occurring; however, H5N1 is still present, replicating, and being shed, leading to the infection of others. A cytokine storm is particularly dangerous for individuals of military age, as evidenced by the large number of soldiers who died during the 1918 influenza epidemic. If large numbers of sub-Saharan soldiers suffer a similar fate from avian influenza, then military and political instability could develop.     

肺鳞癌PD-L1共表达基因及转录调控网络分析

目的 绘制肺鳞癌中细胞程序性死亡配体1(PD-L1)共表达基因关系网络,筛选潜在的PD-L1协同检测生物标志物,寻找可能参与PD-L1调控肿瘤免疫状态的基因及通路.方法 采用TCGA肺鳞癌数据集,利用cBioPortal数据分析平台进行共表达基因集检索,从全基因转录组水平对PD-L1的共表达基因进行Venny分析,筛选目标基因,进行多种类型的聚类和分子关系网络分析,并建立PD-L1调控网络.结果 共获得PD-L1中等程度表达相关基因126个,主要为质膜定位型基因,功能集中在免疫调节过程,其中IRF2/NFKB 1/IRF1三个转录因子参与了对PD-L1基因集30％以上基因的转录调控.经过PD-L1共表达基因集的核心分子筛选,分析网络节点连接度,得到具有最高连接度的前6个节点基因,依次为IFNG、JAK2、STAT1、CTLA4、CD80和CCR5,对这些基因不同表达水平的肺癌患者的生存情况进行分析,结果显示CCR5是一个有意义的预后指标分子.进一步对PD-L1和CCR5的表达谱数据进行相关性分析,结果显示CCR5与PD-L1表达谱水平的Pearson相关系数为0.47(P＜0.05);GO-BP聚类分析显示,CCR5的功能主要聚集在免疫调控、T细胞调控、细胞信号转导等领域,与PD-L1调控网络的功能相符.结论 PD-L1共表达基因集内的主要节点均为免疫相关分子,其中IFNG、CCR5、NFKB1分子对PD-L1共表达基因网络具有最广泛的调控作用,该发现为肺鳞癌免疫治疗的研究和应用奠定了一定基础.     

MR-revealed myelination in the cerebral corticospinal tract as a marker for Pelizaeus-Merzbacher's disease with proteolipid protein gene duplication

BACKGROUND AND PURPOSE: Pelizaeus-Merzbacher's disease (PMD) is caused by mutations in the proteolipid protein (PLP) gene. Recent studies have shown that an increased PLP dosage, resulting from total duplication of the PLP gene, invariably causes the classic form of PMD. The purpose of this study was to compare the MR findings of PMD attributable to PLP duplication with those of PMD arising from a missense mutation. METHODS: Seven patients with PMD, three with a PLP missense mutation in either exon 2 or 5 (patients 1-3), and four with PLP duplication (patient 4 having larger PLP duplication than patients 5-7) were clinically classified as having either the classic or connatal form of PMD. Cerebral MR images were obtained to analyze the presence of myelination and T1 and T2 shortening in the deep gray matter. Multiple MR studies were performed in six of the seven patients to analyze longitudinal changes. RESULTS: Four patients (patients 1-4) were classified as having connatal PMD, whereas the other three (patients 5-7) were classified as having classic PMD. Myelination in the cerebral corticospinal tract, optic radiation, and corpus callosum was observed in three cases of classic PMD with PLP duplication. In patient 4, myelination extended to the internal capsule, corona radiata, and centrum semiovale over a 3-year period. No myelination was observed in three PMD cases with a PLP point mutation. T2 shortening in the deep gray matter was recognized in all patients with PMD. CONCLUSION: The presence of myelination in the cerebral corticospinal tract with diffuse white matter hypomyelination on MR images could be a marker for PMD with PLP duplication. It is suggested that progression of myelination may be present in connatal PMD with large PLP duplication.     

1例早期HIV感染者外周血抗体变化和病毒基因序列分析

目的: 观察HIV早期感染者体内的抗体变化特征,为进一步研究HIV感染早期病毒与宿主的相互作用奠定基础.方法:对一名HIV感染者进行流行病学调查、外周血血清学检测、病毒载量和CD4细胞数检测、体外病毒分离,观察该感染者体内免疫学反应及病毒复制特点.使用巢式PCR方法扩增病毒env区C2-V3区基因,通过测序及序列分析观察病毒部分膜区基因特点.结果: 该感染者为早期感染,外周血抗体S/CO值呈逐渐增高趋势,外周血中针对HIV抗原的特异性条带呈逐渐增多趋势,感染183 d后出现针对所有HIV抗原的特异性抗体,病毒载量在感染后3个月时最高(18 000 cp/ml),随后下降,从该感染者外周血中分离的病毒毒力较强.基因序列分析显示该病毒为CRF01_AE亚型,与泰国93TH054毒株基因距离最近.结论:该感染者体内病毒可能起源于泰国毒株,对HIV早期感染的鉴定为研究我国HIV早期感染者体内病毒进化奠定了基础.     

The role of F-18 FDG PET/CT in evaluating the impact of HIV infection on tumor burden and therapy outcome in patients with Hodgkin lymphoma

Background

To evaluate the impact of HIV infection on tumor burden and therapy outcome following treatment with chemotherapy in patients with Hodgkin lymphoma.

Methods

A total of 136 patients with classical Hodgkin lymphoma were studied (mean age ± SD = 32.31 ± 1.39 years, male = 86, female = 50). Advanced disease (stage III and IV) was present in 64% of patients. HIV infection was present in 57 patients while 79 patients were HIV-negative. Baseline F-18 FDG PET/CT was obtained in all patients. SUVmax, MTV and TLG were determined on the baseline scan to evaluate for tumor burden. All patients completed a standard regimen of adriamycin, bleomycin, vinblastine and dacarbazine (ABVD). After a median period of 8 weeks (range = 6 to 17 weeks), a repeat F-18 FDG PET/CT scan was obtained to evaluate response to therapy using Deauville 5-point scoring system.

Results

The HIV-positive and HIV-negative groups were similar with regards to age and disease stage. The groups were heterogeneous with respect to gender (p = 0.029). The SUVmax, MTV and TLG of lesions were not significant different between the two groups. Complete response was seen in 72.8% of the study population. Presence of HIV infection was associated with higher rate of treatment failure with 40.4% of the HIV-positive patients having treatment failure while only 17.7% of the HIV-negative patients had treatment failure (p = 0.0034). HIV infection was a significant predictor of response to chemotherapy. Effects of SUVmax, MTV, TLG and Ann Arbor stage of the disease were not statistically significant as predictors of therapy outcome. In a multiple logistic regression, presence of HIV infection still remained an independent predictor of therapy outcome in the presence of other factors such as SUVmax, MTV, TLG and the Ann Arbor stage of the disease.

Conclusions

HIV infection is not associated with a higher tumor burden in patients with Hodgkin lymphoma. HIV infection is, however, a strong predictor of poor therapy outcome in patients treated with standard regimen of ABVD.

     

Aerobic exercise training as a potential source of natural antibodies protective against human immunodeficiency virus‐1

Despite the effectiveness of HAART in controlling HIV‐1 replication, the emergence of drug‐resistant viruses in infected patients and the severe side effects caused by the currently used drug regimens and the lack of an effective vaccine necessitate the continued search for new therapeutic strategies for prevention and therapy of HIV disease. Previously we reported that natural autoantibodies, recognizing peptide FTDNAKTI (peptide NTM1) derived from the C2 domain of HIV‐1 gp120, contribute to the control of HIV disease. Here we demonstrated that sera from well‐trained athletic (HIV‐negative) subjects showed high reactivity with peptide NTM1. This result confirms that aerobic exercise training stimulates production of natural autoantibodies, which recognize peptide NTM1. Bioinformatics analysis indicates that these natural autoantibodies could slow down disease progression by blocking the superantigenic site on HIV‐1 gp120. The results suggest that aerobic exercise training may be a promising non‐toxic and inexpensive adjunctive anti‐HIV therapy.     

Knowledge and risks of human immunodeficiency virus transmission among veterans with severe mental illness

This study is among the first to examine knowledge about human immunodeficiency virus (HIV) and behavioral risks for HIV transmission among veterans with severe mental illness (SMI), a group at high risk for HIV infection. This study examined associations between accuracy of HIV knowledge, risk behaviors, and clinical and demographic characteristics in a sample of male veteran psychiatric inpatients diagnosed with SMI (N = 353). Results showed high rates of inaccurate HIV knowledge, with > 40% of patients demonstrating some inaccuracies, particularly those related to the progression and symptoms of acquired immunodeficiency syndrome. Inaccurate HIV knowledge was associated with older age, minority status, education level, marital status, no homelessness within the previous 6 months, and no reported history of illicit intranasal drug use. There is a need for more effective HIV prevention interventions for persons with SMI.     

Colorectal carcinoma metastases: detection with In-111-labeled monoclonal antibody CCR 086

A phase I/II clinical trial with indium-111-labeled antimucin murine monoclonal antibody (MoAb) CCR 086 was conducted. Seventeen patients with histologically proved colorectal carcinoma and known metastatic disease underwent external scintigraphy after administration of 5.5 mCi (203.5 MBq) of In-111 CCR 086 at doses of 5 and 20 mg. Of 25 known lesions, 17 were detected (sensitivity, 68%). The smallest detected lesion in the lung was 1 cm and in the liver was 1.5 cm. The serum half-life of In-111-labeled CCR 086 MoAb was approximately 64 hours. The formation of human antimouse antibody (HAMA) was detected in the serum of four of five patients who received 20 mg of MoAb. No HAMAs were detected in four patients receiving 5 mg of MoAb. No side effects were encountered. Because of effective detection of liver and lung metastases with lower doses (5-20 mg) of CCR 086 conjugated with In-111, further investigations are warranted to assess clinical and therapeutic potentials of CCR 086 in the management of colorectal cancer.