Probable implication of mutations of the X open reading frame in the onset of fulminant hepatitis B

A pathogenic role of precore-defective mutation in the onset of fulminant hepatitis B has been suggested. However, precore-defective mutants do not always cause fulminant hepatitis B and are not always isolated from affected patients. These findings strongly suggest the presence of some additional important mutations outside the precore region in fulminant hepatitis. In the present investigation an attempt was made to sequence the X open reading frame of hepatitis B virus DNA isolated from seven patients with fulminant hepatitis B and five patients with acute hepatitis B. The latter were used as controls. Since the X open reading frame encodes the X protein and contains the core promoter/enhancer II complex, some critical mutations may enhance or disrupt the replication and expression of hepatitis B virus DNA leading to fulminant hepatitis. A C-to-T substitution was found at nucleotide (nt) 1655, an A-to-T substitution at nt 1764 and a G-to-A substitution at nt 1766 in 4, 5 and 5 patients, respectively, out of the seven with fulminant hepatitis. These substitutions were not recognized in the patients with acute hepatitis. These mutations might change the function of the X protein and core promoter/enhancer II complex. It is suggested, therefore, that these mutations, as well as the precore-defective mutation, may play an important role in the pathogenesis of fulminant hepatitis. © Wiley-Liss, Inc.     

Changes in hypothalamic temperature of rats after daily exposure to heat at a fixed time

Rats were subjected to an ambient temperature (T_a) of 33°C for ca. 5 h during the last half of the dark phase for 5, 14 or 28 consecutive days (heat-exposed rats, HE), while control rats were kept at a constant T_a of 24°C. After the heat exposure schedule, the levels of hypothalamic temperature (T_hy) as an index of body core temperature in the HE were significantly lower than those of the controls for 2–4 h in the last half of the dark phase. The low levels of T_hy persisted during the specific period for 1, 3 and 6 days after the end of the 5-, 14- and 28-day heat exposure schedules, respectively. These results confirm that, in rats subjected to daily heat exposure for ca. 5 h at a fixed time per day, their T_hy falls during the period when the rats were previously exposed to heat, and suggest that the duration of the specific T_hy change observed after completing the heat exposure schedule depends on the length of the heat exposure schedule.     

External fixation of unstable pelvic ring fractures: comparative rigidity of some current frame configurations

The effectiveness of external fixation in the stabilisation of pelvic ring fractures was studied in a laboratory cadaveric series. Shearing displacements occurring at sacroiliac joint and symphysis pubis dislocation sites, due to simplified longitudinal loading of the sacrum in an Instron unit, were monitored using variable-impedence transducers. The rigidity of fixation was compared for the Slätis and the Bonnel single anterior frames, for coupled and uncoupled double anterior frames, and for combined anterior-plus-posterior fixation achieved with separate transfixation pin clusters, with through-and-through pin clusters, or with a posterior screw plate. The data showed that the use of posterior fixation provided greatly enhanced stabilisation compared to that achieved with anterior fixation alone. The complex double anterior frames performed only slightly better than did the simpler single anterior frames. In no case, however, was it possible to recover rigidity levels approaching those of the intact pelvis. The results suggest that the transfixation pin arrangement is the most important determinant of pelvic fixation stability, and that further investigation of posterior screw-plate fixation is warranted.     

丙型肝炎病毒核心蛋白在HepG2细胞中的表达及其对端粒酶活性的影响

目的 建立HCV核心蛋白细胞表达模型，并探讨其对细胞端粒酶活性的影响。方法 用PCR法扩增出HCV核心基因cDNA，将其插入真核表达载体pBK-CMV的HindⅢ和BamHⅠ位点间，构建重组质粒pBK-HCVc。再将重组质粒pBK-HCVc和空载体分别导入肝癌细胞株HepG2中，G418筛选，RT－PCR、免疫组化和蛋白印迹鉴定HCV核心蛋白表达。PCR－ELISA法检测端粒酶活性。结果 构建的pBK-HCVc质粒在HepG2细胞中有稳定表达。表达HCV核心蛋白的细胞HepG2-C的端粒酶活性较转染空载体的细胞HepG2-CMV明显升高。结论 HCV核心蛋白上调了端粒酶活性，可能是HCV诱发肝细胞癌的一种途径。     

手术治疗46例胸腰椎爆裂骨折的临床分析

目的探讨前路、后路、前后路联合手术治疗胸腰椎爆裂骨折的特点。方法依据对患者所选择的前路、后路、前后路联合手术术式分为三组,进行影像学评价和神经功能评价。结果各组末次随访脊髓功能评价分级提高情况采用R×C表x2检查方法进行统计学分析,各组差异无显著意义。各组术后cobb氏角改善率应用秩和检验方法进行统计学分析,各组差异无显著意义。结论我们综合应用Denis和McAfee的分型,结合了骨折形态、损伤机制和稳定性评价,对胸腰椎爆裂骨折的治疗有较好的指导意义。手术方式的选择更多基于脊柱机械性稳定性、神经性稳定性评价。综合考虑骨折部位、骨折后时间、患者年龄、工种以及术者对入路的熟悉程度等。     

Molecular and serological evaluation of surface antigen negative hepatitis B virus infection in blood donors from Venezuela

Surface antigen negative hepatitis B virus (HBV) infection was evaluated in Venezuela, by molecular characterization of blood samples positive for antibodies to core antigen (anti-HBc) and negative for surface antigen (HBsAg) in blood donors (residual infections). HBV DNA was found in 11/258 samples (4.3%), and was significantly associated with high levels of anti-HBc antibodies (>25 UI/ml, P < 0.05), while no correlation was found between the presence of HBV DNA and the levels of anti-HBs. Synonymous and non-synonymous mutations were found in the HBV surface region (but not vaccine escape mutants) and in the precore/core region (precore mutants in 2/7 samples and 33-45 bp deletions near the N-terminal core region in 4/19 samples). While HBV genotype F prevails among HBsAg positive samples from blood donors in Venezuela, residual infection isolates were mainly genotypes A and D. Phylogenetic analysis of viral surface and core region revealed discrepancies in genotype designation in 6/9 samples, suggesting the presence of mixed infection or recombination. In conclusion, HBV residual infection in Venezuela does not seem to be frequently observed in HBV genotype F. This type of infection is frequently associated with variants exhibiting mutations in the surface gene that might be affecting the correct recognition by commercial tests, with precore mutants and with core internal deletions. These variants do not seem to cause severe liver disease, and on the contrary, were found circulating at low viremia.     

Transcription from the gene encoding the herpesvirus entry receptor nectin-1 (HveC) in nervous tissue of adult mouse

Hepatitis C virus core protein, in addition to being a component of the viral capsid, has a number of regulatory functions. Here we showed two bodies of evidence indicating that a fraction of the core protein species is a substrate of the ubiquitin (Ub)-proteasome pathway of targeted proteolysis. First, the core protein processing the C-terminal hydrophobic region is metabolically unstable, and incubation with a proteasome inhibitor led to a significant accumulation of the protein. Second, an in vivo ubiquitylation assay indicates conjugation of multi-Ub chain to the unstable core protein. In contrast, a stable form of core protein, p21, is also able to be ubiquitylated, but it links to a single or only a few Ub moiety. Therefore, processing event(s) at the C-terminal hydrophobic domain of HCV core protein may affect the ubiquitylation pathway, particularly the efficiency of the multi-Ub chain assembly, resulting in stable, matured core proteins.     

蛋白激酶R与丙型肝炎病毒核心蛋白相互作用区域定位

目的： 观察丙型肝炎病毒（HCV）核心蛋白（CP）对蛋白激酶R(PKR)表达的影响；定位PKR与CP直接结合的区域。方法: 对Huh-7、转染表达CP的Huh-7及含有全长HCV复制子(replicon) Huh-7细胞株的PKR表达水平及干扰素（IFN）诱导前后replicon Huh-7细胞中HCV结构蛋白和非结构蛋白表达水平作比较；对CP与PKR进行免疫共沉淀试验、谷胱苷肽S转移酶（GST）结合试验。结果: Replicon Huh-7中PKR表达水平高于Huh-7及转染表达CP的Huh-7；IFN诱导后PKR表达增加，且明显抑制HCV结构和非结构蛋白的表达；PKR能与CP直接结合，依赖于PKR的N端1-180氨基酸（aa）。结论: CP能直接作用于PKR N端1-180 aa，导致PKR组成性激活，从而干扰PKR介导的相关信号转导通路。CP与PKR的相互作用是HCV病毒蛋白与细胞蛋白相互作用又一新的模式，在HCV持续感染及肝癌2者发病机制方面可能起重要作用。     

Significance of the latissimus dorsi for shoulder instability. I. Variations in its anatomy around the humerus and scapula

In a cadaveric instability model that leaves all muscles intact initially, we studied anteroinferior glenohumeral dislocation behavior after section of the ligaments on the humeral side of the joint. In this study, the latissimus dorsi seemed to play a role when complete section did not result in a locked anteroinferior dislocation. We therefore initiated a study to test the hypothesis that the latissimus dorsi may, in certain circumstances, depending on variations in its anatomy, influence dislocation behavior. Here, in Part I, we present the results of the anatomic study of latissimus dorsi and its tendons. The anatomy of the latissimus dorsi pertaining to the scapula and humerus was studied in 100 cadaver specimens. The distance between the uppermost part of the tendon of both the latissimus dorsi and the teres major and the edge of the articular cartilage of the humeral head (tendon-cartilage distance, TCD) as well as the width and length of the tendons were measured. Furthermore, the relationship between latissimus dorsi and the inferior angle of the scapula was studied. The tendon of the latissimus dorsi inserted at a variable distance from the cartilage of the humeral head: the TCD ranged from 12.6 to 31.6 mm (mean 21.06 mm+/-5.11 mm). The latissimus dorsi can have muscular fibers arising from the inferior angle of the scapula (type 1 scapular connection, 43%). Alternatively, there may be only a few fibrous strands between the muscle and the scapula or there may be an intervening bursa (type 2 scapular connection, 57%). This variability in the morphology of the latissimus dorsi may be a factor explaining the differences observed in a study of humerus-based sequential cutting of the glenohumeral capsule. This possibility is explored in Part II of the study. The latissimus dorsi may also complete the tendinous protection of the humeral side of the capsule generally provided by the rotator cuff.     

Cardiac ankyrin repeat protein is preferentially induced in atrophic myofibers of congenital myopathy and spinal muscular atrophy

Cardiac ankyrin repeat protein (CARP), which is structurally characterized by the presence of four ankyrin repeat motifs in its central region, is believed to be localized in the nucleus and to participate in the regulation of cardiac-specific gene expression in cardiomyocytes. However, we recently found that CARP was induced in skeletal muscle by denervation, leading us to speculate that CARP may be induced under some pathological conditions. In the present study, we immunohistochemically analyzed the expression of CARP in 11 cases of spinal muscular atrophy (SMA) and 14 cases of congenital myopathy. In SMA, CARP was expressed selectively in severely atrophic myofibers, suggesting that CARP expression may reflect the status of muscle atrophy. Furthermore, in the congenital myopathies, the expression patterns of CARP were distinct among the subtypes, which included nemaline myopathy, myotubular myopathy, central core disease, and congenital fiber type disproportion. Although CARP was preferentially expressed in severely damaged myofibers in nemaline myopathy, it was not detected in central core disease. These findings suggest that immunohistochemical evaluation of CARP may be helpful in the diagnosis of SMA and the congenital myopathies.