Repair of DNA lesion O 6-methylguanine in hepatocellular carcinogenesis

Evidence from both experimental carcinogenesis and studies in human cirrhotic liver suggest that defective repair of the promutagenic DNA base lesion, O ⁶-methylguanine, is a factor in the multistep process of hepatocellular carcinogenesis. Ubiquitous environmental alkylating agents such as N-nitroso compounds can produce O ⁶-methylguanine in cellular DNA. Unrepaired, O ⁶-methylguanine can lead to the formation of G ? A transition mutations, a known mechanism of human oncogene activation and tumour suppressor gene inactivation. Combined treatment of rodents with an agent producing O ⁶-methylguanine in DNA, and an agent promoting cell proliferation, leads to development of hepatic nodules and hepatocellular carcinoma (HCC), cell division, hence DNA replication, being required for the propagation of tumorigenic mutation(s) in hepatocyte DNA. The paramount importance of O ⁶-methylguanine in hepatocellular carcinogenesis is indicated by the observation that transgenic mice engineered to have increased hepatic levels of repair enzyme O ⁶-methylguanine-DNA methyltransferase (MGMT) are significantly less prone to hepatocellular carcinogenesis following alkylating agent treatment. Cirrhosis is a universal risk factor for development of human HCC, and a condition that is characterized by increased hepatocyte proliferation as a result of tissue regeneration. Levels of the human repairing enzyme for O ⁶-methylguanine were found to be significantly lower in cirrhotic liver than in normal tissue. In accord with findings from animal models, this suggested a mechanism in which persistence of O ⁶-methylguanine due to defective DNA repair by MGMT, together with increased hepatocyte proliferation, might lead to specific gene mutation(s) and hepatocellular carcinogenesis. Screening for the presence and persistence of O ⁶-methylguanine in human DNA presently involves formidable technical difficulty. Indications are that such limitations might be overcome by the use of an ultrasensitive method such as immuno-polymerase chain reaction (PCR). This approach should allow parallel measurement of DNA adduct and repair enzyme in routine liver biopsy samples. It might also enable investigation of O ⁶-methylguanine in human genes specifically associated with hepatocellular carcinogenesis. Given the wide variation in human MGMT levels observed between individuals, tissues, and cells, this technology should be adapted to permit the ultrasensitive localisation and measurement of adducts and repairing enzyme in liver biopsy tissue sections. Ability to ultrasensitively measure O ⁶-methylguanine, and its repair enzyme, should prove valuable in the risk assessment of cirrhotic patients for developing hepatocellular carcinoma. Received for publication on July 6, 1998; accepted on Aug. 12, 1998     

Comparative studies on cytotoxic and genotoxic effects of two organic mercury compounds in lymphocytes and gastric mucosa cells of sprague-dawley rats

Human lymphocytes (HL) as well as lymphocytes (RL), hepatocytes (RH), and gastric mucosa cells (GM) of Sprague-Dawley rats were treated in vitro for 1 h with methylmercury chloride (MMC, 0.5–4 μg/ml) and dimethylmercury (DMM, 5–40 μg/ml). The cytotoxicity of the two organic mercury compounds was assessed by dye exclusion, and the extent of induced DNA fragmentation was measured with a single-cell microgel electrophoresis assay. Both MMC and DMM induced DNA damage and cytotoxicity in a dose-related manner in HL, RL, and GM. MMC was more effective in causing a significant increase in median DNA migration than DMM at doses yielding approximately the same degree of cytotoxicity. In rat hepatocytes the MMC-induced DNA damage was, however, lower than in the other cells. An analysis of repair kinetics following exposure to 2 μg/ml MMC was carried out in human lymphocytes obtained from an adult male donor. The bulk of DNA repair occurred 90 min after in vitro exposure, and it was about complete by 120 min following cessation of exposure. Finally, in order to have a basis for extrapolating to the human situation, in vivo studies were performed with Sprague-Dawley rats, also assessing the DNA damage and cytotoxicity in the lymphocytes and gastric mucosa cells. These in vivo results after oral exposure may be directly compared to the in vitro data obtained in the same cells. © 1993 Wiley-Liss, Inc.     

Long-term result and current status of the Lichtenstein open tension-free hernioplasty

Summary The tension-free hernioplasty project began in 1984 at the Lichtenstein Hernia Institute. The method consists of complete reinforcement of the inguinal floor with a large sheet of mesh, with adequate mesh tissue interface beyond the boundary of the inguinal floor and creation of a new internal ring made of prosthesis. The preliminary report of this operation was published in 1989, with no recurrence at that point in time. Shortly after the submission of the report, several recurrences were encountered. Based on the lesson learned from those recurrences, the operation was slightly modified and reported in 1991 [Amid 1993]. Since then, the Lichtenstein technique has gained world-wide popularity. Outcome measures identical to ours and other authors have been achieved by even those surgeons who have no special interest or expertise in herniology. The purpose of this article is to report the current state of the open tension-free hernioplasty for the repair of primary and recurrent inguinal hernias.     

髓芯减压联合VEGF与胶原基骨移植材料对兔股骨头缺血坏死的修复作用

探讨髓芯减压联合血管内皮生长因子（VEGF）与胶原基骨移植材料对兔股骨头缺血坏死的修复作用。方法 对24只SPF级家兔股骨头内注射无水乙醇建立兔股骨头坏死模型,然后将家兔随机分为模型对照组（A组）、髓芯减压+自体松质骨组（B组）、髓芯减压+胶原基骨修复材料组（C组）和髓芯减压+胶原基骨修复材料+VEGF组（D组）,每组6只,共治疗12周。通过苏木精伊红（HE）染色观察组织切片形态。采用骨密度分析系统（QCT PRO V6.1）测量家兔股骨头骨密度(BMD)。Western blot检测VEGF、Collagen I、Runt相关转录因子2（RUNX2）、骨钙素（OCN）、Wnt-3a、β-catenin和GSK-3β的蛋白表达。结果 术后12周时,与B、C组比较,D组家兔股骨头骨小梁排列较整齐,骨髓中观察到大量微血管的形成,可见明显成骨,且坏死区基本被修复。与A组相比,B组、C组和D组家兔股骨头空骨陷窝比率均显著降低(P＜0.05);D组家兔股骨头空骨陷窝比率小于B组和C组(P＜0.05)。与A组相比,B组、C组和D组家兔股骨头的骨密度均显著升高(P＜0.05);D组骨密度显著高于B组和C组(P＜0.05)。与B组和C组相比,D组VEGF、Collagen I、RUNX2和OCN的蛋白表达水平均显著升高（P＜0.05）;与B组和C组相比,D组Wnt-3a和β-catenin的蛋白表达水平均显著升高,GSK-3β的蛋白表达水平显著降低（P＜0.05）。结论 髓芯减压联合VEGF与胶原基骨移植材料对兔股骨头缺血坏死可有效促进坏死股骨头的修复,提高骨密度及成骨蛋白的表达     

Evolution of Vertebroplasty: A Biomechanical Perspective

This paper is a collection of computational, finite element studies on vertebroplasty performed in our laboratory, which attempts to provide new biomechanical evidence and a fresh perspective into how the procedure can be implemented more effectively toward the goal of preventing osteoporosis-related fractures. The percutaneous application of a bone cement to vertebral defects associated with osteoporotic vertebral compression fracture has proven clinical successful in alleviating back pain. When the biomechanical efficacy of the procedure was examined, however, vertebroplasty was found to be limited in its ability to provide sufficient augmentation to prevent further fractures without risking complications arising from cement extravasations. The procedure may instead be more efficient biomechanically as a prophylactic treatment, to mechanically reinforce osteoporotic vertebrae at risk for fracture. Patient selection for such intervention may be reliably achieved with the more accurate fracture risk assessments based on vertebral strength, predicted using geometrically detailed, specimen-specific finite element models, rather than on bone density alone. Optimal cement volume, placement, and material properties were also recommended. The future of vertebroplasty involving biodegradable augmentation material laced with osteogenic agents that upon release will stimulate new bone growth and increase bone mass was proposed.     

脊髓损伤后神经修复过程中的细胞微环境北大核心

背景:以往的研究显示单一改变脊髓损伤区域某一基因表达或者某一细胞的状态,对脊髓损伤后功能恢复无显著影响,而大量证据表明调控脊髓损伤后紊乱的细胞微环境是神经功能恢复的关键因素。目的:对脊髓损伤前后细胞微环境的生物学特性,包括多种细胞之间的相互调控以及细胞外组分对损伤神经修复的作用和机制进行综述。方法:由第一作者检索PubMed及Web of Science数据库,英文检索词为“spinal cord injury,glial cell,neuron,immune cell,neural stem cell,extracellular matrix,cytokine,extracellular vesicle,regeneration”。文献检索的时间范围为2000年1月至2021年12月,最终筛选出64篇文献进行分析。结果与结论:①脊髓损伤后,在细胞微环境的细胞组分中,占比最高的胶质细胞间的相互作用,以及与神经元的相互调控作用最为关键。②在脊髓损伤后的细胞外组分中,利用生物相容性良好的水凝胶模仿天然细胞外基质,可有效模拟和重建损伤区域内的细胞微环境,促进轴突伸长。③在脊髓损伤后的细胞外调节因子中,促炎因子如肿瘤坏死因子α和白细胞介素1β等加剧了细胞微环境的炎症反应,应用受体抑制剂或阻断相关通路抑制上述促炎因子的表达是一种有效的治疗方法,同时在脊髓微环境中增加白细胞介素10等抗炎因子的表达,抑制损伤区域炎症发展的研究也陆续出现。④最近被重视起来的细胞外囊泡作为传递信息的载体在细胞微环境中也发挥了重要作用。⑤文章揭示了脊髓损伤后细胞微环境中的包括细胞组分和细胞外组分之间的多组相互调控关系,证实了细胞微环境中各组分之间所发挥的神经修复作用并不是孤立的。     

Analysis of class switch recombination and somatic hypermutation in patients affected with autosomal dominant hyper-IgM syndrome type 2

Autosomal recessive form of hyper-IgM syndrome type 2 (AR-HIGM2) is secondary to mutations affecting both alleles of AICDA gene encoding activation-induced cytidine deaminase, characterized by defects of immunoglobulin class switch recombination (CSR) and somatic hypermutation (SHM) in most of the patients. We herein report the immunological phenotype of seven patients carrying a single heterozygous R190X mutation in AICDA. Variable defect in in vivo CSR inherited as an autosomal dominant (AD) trait strongly suggests that this heterozygous AICDA mutation causes HIGM (AD-HIGM2). In AD-HIGM2 B cells, CSR was consistently found impaired in vitro. However, in contrast to AR-HIGM2, the CSR-induced double-stranded DNA breaks in the switch region of IgM heavy chain gene were detected. The SHM frequency in V regions of IgM heavy chain gene in B cells was normal in all (but one patient). The characteristics of the AD-HIGM2 phenotype indicate that the AID C-terminal region may be involved in DNA repair machinery required for CSR.     

Early-onset gastric carcinomas display molecular characteristics distinct from gastric carcinomas occurring at a later age

Gastric cancer is thought to result from a combination of environmental factors and accumulation of specific genetic alterations, and consequently mainly affects older patients (>50 years of age). Fewer than 10% of patients present with the disease before 45 years of age and these young patients are thought to develop carcinomas with a different molecular genetic profile from that of sporadic carcinomas occurring at a later age. Forty early-onset gastric carcinoma resection specimens were characterized for microsatellite instability (MSI) and loss of heterozygosity status using 22 polymorphic microsatellite markers. Twenty-four biopsies were additionally evaluated for the presence of MSI. No MSI was observed in any of the cases analysed. Losses were infrequent, but were most common for the D1S234 (26.1%) and D1S1676 (17.4%) markers, flanking the RUNX3 gene; for the p53ALU (23.1%) and TP53 (15.4%) markers, near the TP53 gene; and for the D16S2624 (17.2%) marker, near the E-cadherin (CDH1) gene. All cases with loss of CDH1, as well as 6/7 cases with loss of TP53, displayed aberrant staining of the corresponding proteins, pointing to a functional role for these proteins in early-onset gastric carcinogenesis. No germline CDH1, TP53 or RUNX3 mutations were detected in any of the cases analysed. No correlation was observed between non-functional E-cadherin and the histological type of the tumours analysed. Finally, Epstein-Barr virus was not detected in any of the cases analysed. On the basis of these results, early-onset gastric carcinomas appear to have characteristics distinct from gastric carcinomas occurring at a later age.     

DNA聚合酶β可能参与了更广泛的细胞反应并引起遗传不稳定

近年来有关真核细胞中ＤＮＡ聚合酶的研究迅速增多。 1985年前 ,确认的ＤＮＡ聚合酶只有 3种 :聚合酶α、β以及线粒体聚合酶γ。时至今日已至少发现了 14种ＤＮＡ聚合酶 ,包括聚合酶α、β、γ、δ、ε、ζ、η、θ、ι、κ、λ、μ、ＲＥＶ1及ＭＵＳ30 8,在真核细胞ＤＮＡ的复制、修复、跨损伤合成、ＤＮＡ重组以及细胞周期调控等多种重要的生物代谢过程中发挥重要作用[1] 。在一定程度上 ,细胞内的各种聚合酶均有其独特的功能和作用领域。如具有引物酶活性的聚合酶α的作用是启动ＤＮＡ复制并合成ＲＮＡ -ＤＮＡ引物 ;聚合酶δ和ε是主要…