WIF-1 promoter region hypermethylation as an adjuvant diagnostic marker for non-small cell lung cancer-related malignant pleural effusions

Purpose  Malignant pleural effusion is an important staging criterion in non-small cell lung cancer (NSCLC). Although cytologic examination remains the major diagnostic tool for NSCLC-related malignant pleural effusion, sometimes other invasive methods maybe required. Aberrant activation of Wnt signaling pathway due to Wnt inhibitory factor-1 (WIF-1) promoter region hypermethylation is common in NSCLC, and can be specifically detected by methylation-specific polymerase chain reaction (MSP). We hypothesized that WIF-1 promoter region MSP can be used to improve the diagnostic yield of NSCLC-related malignant pleural effusion. Methods  We performed WIF-1 promoter region MSP in 36 definite malignant pleural effusions from consecutive NSCLC patients and 35 pleural effusion specimens of benign origin. Pleural effusion cells were collected for DNA extraction. After bisulfite treatment, DNA was amplified by methylation-specific and unmethylation-specific primers, respectively, to identify the methylation status of WIF-1 promoter region. Results  The results of WIF-1 promoter region MSP were positive in 25 (69.4%) of 36 NSCLC patients with malignant pleural effusion. In addition, the results of WIF-1 promoter region MSP were negative in all 35 patients with pleural effusion of benign origin. The age, gender, and smoking status of patients were not correlated with the methylation status of WIF-1 promoter region in NSCLC-related malignant pleural effusion. Conclusions  WIF-1 promoter region MSP might be used as an adjuvant tool to complement cytologic examination for the diagnosis of NSCLC-related malignant pleural effusion. T.-M. Yang and S.-W. Leu are the first authors and contributed equally.     

A readapted Malone schistosome transmission index model

Schistosome transmission index can be directly used to give an overall evaluation and prediction of schistosomiasis transmission through a mathematical model. In the light of the mechanism of schistosomiasis transmission process, an improved Malone schistosome transmission index model was established based on the previous study of survivor rate of the snail under extreme temperatures by Hong Qingbiao. Meteorological data from FAO and Institute of Geographic Sciences and Resources Research, CAS was used to calculate the schistosome transmission index by the new model in the schistosomiasis epidemic region of China, and the result was re-classified into transmission index classes. The new model changed the class of “epidemic area” into that of “non-epidemic area” in the regions of the southern Henan province, the northern Anhui province and the middle Jiangsu province by transmission index threshold of 900. The geographic boundary between the epidemic and the non-epidemic regions was found fitly to coincide with the −2 °C isotherm of January in China, whereas the underpinning cause needs to be further explored in the future. Finally discussed was the calculation result of this meteorological model having some difference from actual epidemiology as more factors of relevance could be considered to be incorporated into the model, also discussed were a few questions concerning the model itself and the possible future improvement directions.     

复合髂腹下／髂腹股沟神经阻滞对小儿七氟烷麻醉后苏醒期的影响

目的观察髂腹下／髂腹股沟神经阻滞对门诊腹股沟区手术小儿七氟烷麻醉后苏醒期的影响。方法门诊拟行择期单侧腹股沟区手术的小儿60例,随机分为2组,组Ⅰ为全身麻醉组（n=30）,组Ⅱ为全身麻醉联合INB组（n=30）。两组患儿均吸入七氟烷、笑气麻醉诱导,插入喉罩并保持自主呼吸。组Ⅰ患儿静脉注射芬太尼2～4μg·kg^-1,组Ⅱ患儿未使用静脉镇痛药,由同一麻醉医生实施骼腹股沟／髂腹下神经联合阻滞。两组患儿术中均以七氟烷、笑气维持麻醉。观察并记录：麻醉时间;入PACU后10min时的疼痛评分;PACU的滞留时间;PACU内发生躁动的患儿例数;局部麻醉部位有无血肿等并发症。结果入PACU后10min时组Ⅱ患儿的疼痛评分2．4±1．8,较组Ⅰ患儿的4．3±2．2为低,差异具有统计学意义,P〈0．05。组Ⅰ有6例,组Ⅱ有2例患儿发生了的严重的疼痛。组间比较差异无统计学意义。P〉0．05。组Ⅱ有2例（6．7％）患儿发生了躁动,较组Ⅰ10例（33．3％）为少,差异具有统计学意义。组Ⅱ患儿PACU的滞留时间为25±4min较组Ⅰ的31±3min为短,组间比较差异具统计学意义,P〈0．05。结论全麻联合髂腹下／髂腹股沟神经阻滞可为门诊行腹股沟区手术的小儿提供良好的镇痛并可降低七氟烷麻醉后EA的发生率,有利于此类小儿的围术期安全。     

乳腺癌根治术后局部区域复发再手术26例体会

目的乳腺癌根治术后局部或邻近区域复发转移经再次手术治疗26例取得较好的临床效果。方法 26例乳腺癌患者中原改良根治20例,标准根治6例。结果随访26例胸部及邻近区域复发再手术病例中,两年内死亡6例,生存3～5年9例,生存5年以上11例,对乳腺癌术后局部或邻近区域复发的再手术治疗,能明显提高生存年限。结论作者认为局部＂扩大手术切除＋综合辅助治疗＂对局部复发病情控制优于其他治疗方法。     

Brain distribution of genes related to changes in locomotor activity

The relationship between genes and behavior, and particularly the hyperactive behavior, is clearly not linear nor monotonic. To address this problem, a database of the locomotor behavior obtained from thousands of mutant mice has been previously retrieved from the literature. Data showed that the percent of genes in the genome related to locomotor hyperactivity is probably more than 1.56%. These genes do not belong to a single neurotransmitter system or biochemical pathway. Indeed, they are probably required for the correct development of a specific neuronal network necessary to decrease locomotor activity. The present paper analyzes the brain expression pattern of the genes whose deletion is accompanied by changes in locomotor behavior. Using literature data concerning knockout mice, 46 genes whose deletion was accompanied by increased locomotor behavior, 24 genes related to decreased locomotor behavior and 23 genes not involved in locomotor behavior (but important for other brain functions) have been identified.These three groups of genes belonged to overlapping neurotransmitter systems or cellular functions. Therefore, we postulated that a better predictor of the locomotor behavior resulting from gene deletion might be the brain expression pattern. To this aim we correlated the brain expression of the genes and the locomotor activity resulting from the deletion of the same genes, using two databases (Allen Brain Atlas and SymAtlas). The results showed that the deletion of genes with higher expression level in the brain had higher probability to be accompanied by increased behavioral activity. Moreover the genes that were accompanied by locomotor hyperactivity when deleted, were more expressed in the cerebral cortex, amygdala and hippocampus compared to the genes unrelated to locomotor activity. Therefore, the prediction of the behavioral effect of a gene should take into consideration its brain distribution. Moreover, data confirmed that genes highly expressed in the brain are more likely to induce hyperactivity when deleted. Finally, it is suggested that gene mutations linked to specific behavioral abnormalities (e.g. inattention) might probably be associated to hyperactivity if the same gene has elevated brain expression.     

In vivo analysis of a new R5 tropic SHIV generated from the highly pathogenic SHIV-KS661, a derivative of SHIV-89.6

Although X4 tropic SHIVs have been studied extensively, they show distinct infection phenotypes from those of R5 tropic viruses, which play an important role in HIV-1 transmission and pathogenesis. To augment the variety of R5 tropic SHIVs, we generated a new R5 tropic SHIV from the highly pathogenic X4 tropic SHIV-KS661, a derivative of SHIV-89.6. Based on consensus amino acid alignment analyses of subtype B R5 tropic HIV-1, five amino acid substitutions in the third variable region successfully changed the secondary receptor preference from X4 to R5. Improvements in viral replication were observed in infected rhesus macaques after two passages, and reisolated virus was designated SHIV-MK38. SHIV-MK38 maintained R5 tropism through in vivo passages and showed robust replication in infected monkeys. Our study clearly demonstrates that a minimal number of amino acid substitutions in the V3 region can alter secondary receptor preference and increase the variety of R5 tropic SHIVs.     

Prevalence of hepatitis B virus MHR mutations and their correlation with genotypes and antiviral therapy in chronically infected patients in Serbia

Understanding the prevalence and diversity of HBsAg variants in a population is fundamental to assay design and planning vaccination programs. It has been shown that mutations within the S gene, caused by selection or natural variation, can lead to false‐negative results in assays for HBsAg, or have clinical implications, such as evading anti‐HBV immunoglobulin therapy or vaccine‐induced immunity. The region of HBsAg where most of these mutations occur is known as the major hydrophilic region (MHR). The aim of this study was to determine the prevalence and mutational patterns of MHR mutations in patients with chronic hepatitis B, and their correlation with patient characteristics, viral factors and antiviral therapy. The study comprised 164 plasma samples from patients with chronic hepatitis B, of which, 34.8% were on long‐term lamivudine monotherapy. Direct sequencing of part of the S/pol gene was used for identification of HBsAg mutations, HBV genotypes, subgenotypes and HBsAg subtypes. The overall frequency of MHR mutations was 22.6%, but it varied significantly between untreated and treated patients (16.8% vs. 33.3%). The most frequent substitution was at position 120 (9.1%) whereas the most common vaccine‐escape position, 145, was affected in 1.8% of isolates. The presence of MHR mutations was correlated with genotype D, subgenotype D3, and ayw2/ayw3 HBsAg subtypes and to older age (>40 years). It is concluded that natural viral variability present in a geographical region, duration of infection, and antiviral therapy are among the major factors associated with the occurrence of MHR mutations. J. Med. Virol. 82: 1160–1167, 2010. © 2010 Wiley‐Liss, Inc.     

Hepatitis C infection among intravenous drug users attending therapy programs in Cyprus

The most high‐risk population for HCV transmission worldwide today are intravenous drug users. HCV genotypes in the general population in Cyprus demonstrate a polyphyletic infection and include subtypes associated with intravenous drug users. The prevalence of HCV, HBV, and HIV infection, HCV genotypes and risk factors among intravenous drug users in Cyprus were investigated here for the first time. Blood samples and interviews were obtained from 40 consenting users in treatment centers, and were tested for HCV, HBV, and HIV antibodies. On the HCV‐positive samples, viral RNA extraction, RT‐PCR and sequencing were performed. Phylogenetic analysis determined subtype and any relationships with database sequences and statistical analysis determined any correlation of risk factors with HCV infection. The prevalence of HCV infection was 50%, but no HBV or HIV infections were found. Of the PCR‐positive samples, eight (57%) were genotype 3a, and six (43%) were 1b. No other subtypes, recombinant strains or mixed infections were observed. The phylogenetic analysis of the injecting drug users' strains against database sequences observed no clustering, which does not allow determination of transmission route, possibly due to a limitation of sequences in the database. However, three clusters were discovered among the drug users' sequences, revealing small groups who possibly share injecting equipment. Statistical analysis showed the risk factor associated with HCV infection is drug use duration. Overall, the polyphyletic nature of HCV infection in Cyprus is confirmed, but the transmission route remains unknown. These findings highlight the need for harm‐reduction strategies to reduce HCV transmission. J. Med. Virol. 82:263–270, 2010. © 2009 Wiley‐Liss, Inc.     

The ADAMs family: coordinators of nervous system development, plasticity and repair

A disintegrin and metalloprotease (ADAM) transmembrane proteins have metalloprotease, integrin-binding, intracellular signaling and cell adhesion activities. In contrast to other metalloproteases, ADAMs are particularly important for cleavage-dependent activation of proteins such as Notch, amyloid precursor protein (APP) and transforming growth factor alpha (TGFalpha), and can bind integrins. Not surprisingly, ADAMs have been shown or suggested to play important roles in the development of the nervous system, where they regulate proliferation, migration, differentiation and survival of various cells, as well as axonal growth and myelination. On the eleventh anniversary of the naming of this family of proteins, the relatively unknown ADAMs are emerging as potential therapeutic targets for neural repair. For example, over-expression of ADAM10, one of the alpha-secretases for APP, can prevent amyloid formation and hippocampal defects in an Alzheimer mouse model. Another example of this potential neural repair role is the finding that ADAM21 is uniquely associated with neurogenesis and growing axons of the adult brain. This comprehensive review will discuss the growing literature about the roles of ADAMs in the developing and adult nervous system, and their potential roles in neurological disorders. Most excitingly, the expanding understanding of their normal roles suggests that they can be manipulated to promote neural repair in the degenerating and injured adult nervous system.