Early detection of diabetic kidney disease: Present limitations and future perspectives

Diabetic kidney disease(DKD) is one of the most common diabetic complications, as well as the leading cause of chronic kidney disease and end-stage renal disease around the world. To prevent the dreadful consequence, development of new assays for diagnostic of DKD has always been the priority in the research field of diabetic complications. At present, urinary albumin-to-creatinine ratio and estimated glomerular filtration rate(eG FR) are the standard methods for assessing glomerular damage and renal function changes in clinical practice. However, due to diverse tissue involvement in different individuals, the so-called "non-albuminuric renal impairment" is not uncommon, especially in patients with type 2 diabetes. On the other hand, the precision of creatinine-based GFR estimates is limited in hyperfiltration status. These facts make albuminuria and eG FR less reliable indicators for early-stage DKD. In recent years, considerable progress has been made in the understanding of the pathogenesis of DKD, along with the elucidation of its genetic profiles and phenotypic expression of different molecules. With the help of ever-evolving technologies, it has gradually become plausible to apply the thriving information in clinical practice. The strength and weakness of several novel biomarkers, genomic, proteomic and metabolomic signatures in assisting the early diagnosis of DKD will be discussed in this article.

Effects of sex and generation on hepatitis B viral load in families with hepatocellular carcinoma

AIM To explore factors associated with persistent hepatitis B virus (HBV) infection in a cohort of hepatocellular carcinoma (HCC)-affected families and then investigate factors that correlate with individual viral load among hepatitis B surface antigen (HBsAg)-positive relatives. METHODS questionnaire. Demographics, relationship to index case, HBsAg status of mothers and index cases were evaluated for association with the HBV persistent infection or viral load by generalized estimating equation analysis. RESULTS Among 729 relatives enrolled, parent generation (P = 0.0076), index generation (P = 0.0044), mothers positive for HBsAg (P = 0.0007), and HBsAg-positive index cases (P = 5.98 x 10(-8)) were associated with persistent HBV infection. Factors associated with HBV viral load were evaluated among 303 HBsAg-positive relatives. Parent generation (P = 0.0359) and sex (P = 0.0007) were independent factors associated with HBV viral load. The intra-family HBV viral load was evaluated in families clustered with HBsAg-positive siblings. An intra-family trend of similar HBV viral load was found for 27 of 46 (58.7%) families. Male offspring of HBsAg-positive mothers (P = 0.024) and older siblings were associated with high viral load. CONCLUSION Sex and generation play important roles on HBV viral load. Maternal birth age and nutritional changes could be the reasons of viral load difference between generations.

Association of Caveolin-1 polymorphisms with colorectal cancer susceptibility in Taiwan

AIM: To investigate the association of Caveolin-1 (Cav-1) polymorphisms with colorectal cancer (CRC) risk in a central Taiwan Residents population. METHODS: Three hundred and sixty-two patients with colorectal cancer and the same number of recruited age-and gender-matched healthy controls were genotyped. And only those matches with all single nucleotide poly-morphisms data (case/control = 362/362) were selected for final analyzing. RESULTS: There were significant differences between CRC and control groups in the distributions of their genotypes (P = 1.6 × 10 -12 and 3.0 × 10 -4 ) and allelic frequencies (P = 2.3 × 10 -13 and 4.0 × 10 -5 ) in the Cav-1 G14713A (rs3807987) and T29107A (rs7804372) poly-morphisms respectively. As for the haplotype analysis,those who had GG/AT or GG/AA at Cav-1 G14713A/ T29107A showed a 0.68-fold (95% CI: 0.48-0.98) de- creased risk of CRC compared to those with GG/TT,while those of any other combinations were of increased risk. There were joint effects of Cav-1 G14713A and T29107A genotype with smoking status on individual CRC susceptibility. CONCLUSION: This is the first report providing evidence of Cav-1 being involved in CRC and it may be novel useful genomic markers for early detection of CRC.

Effect of verapamil on nitric oxide synthase in a portal veinligated rat model: Role of prostaglandin

AIM： To investigate the effects of verapamil on nitric oxide （NO） synthesis in a portal vein-ligated rat model. METHODS： Systemic and splanchnic hemodynamics were measured by radiolabeled microspheres in portal hypertensive rats after acute administration of verapamil （2 mg/kg） on chronic treatment with N^W-nitro-L-arginine （NNA）（80 mg/kg） and/or indomethacin （2 mg/kg） . RESULTS： Verapamil （2 mg/kg） caused a marked fall in both arterial pressure and cardiac output accompanied by an insignificant change in the portal pressure and no change in portal venous inflow. This result suggested that verapamil did not cause a reduction in portal vascular resistance of portal hypertensive rats, which was similar between N^w- nitro-Loarginine-treated and indomethacin-treated groups. CONCLUSION： In portal hypertensive rats pretreated with NNA and/or indomethacin, acute verapamil administration can not reduce the portal pressure, suggesting that NO and prostaglandin play an important role in the pathogenesis of splanchnic arterial vasodilation in portal hypertension.

A substrate scaffold for assessment of nerve regeneration and neurodegenerative diseases

Neuroregenerafion is a complex topic in neurosci- ence and includes 3 concepts： neurogenesis, neuro- plasticity, and neurorestoration. After injury of the nervous system, axons have the capacity for self-re- pair, regrowth or proliferation. The peripheral ner- vous system is more effective at restoring damaged axons than the central nervous system （CNS）. This is because formation of scar tissue in the CNS in- fluences neural regrowth or synthesis of growth-in- hibiting proteins, thereby preventing reconstruction of a neural circuit （Silver and Miller, 2004; Enciu et al., 2011）. Parkinson＇s disease （PD） and Alzheimer＇s disease （AD） are two most common degenerative diseases of the CNS among the elderly.

Retrospective survey of avian influenza H5N1 infection in Northern Vietnam by using a combinational serologic assay

Outbreaks of highly pathogenic avian influenza H5N1 virus have occurred in Vietnam since 2003. However, how people got avian H5N1 infection in Northern Vietnam is still unclear. We therefore performed a combination of the serologic assays H5N1 ELISA and H5 western blot to detect anti-H5N1 specific antibodies. Sera samples of 149 subjects with suspected H5N1 infection from three provinces of Northern Vietnam were collected from September 2006 to March 2007. Our results indicated that this combinational assay showed high sensitivity (100%) and specificity (95%) when compared with hemagglutinin inhibition (HI) assay. Fifty-one sera samples (34.2%) contained specific antibodies against H5N1 viruses. Poultry raisers (32/77;41.6%) showed higher H5N1 infection rates than slaughterers (12/41;29.3%) and health care workers (7/31;22.6%). Contact history with sick or dead poultry in household or slaughter-house (p < 0.05) and lack of protective equipment use when in contact with dead poultry (p < 0.05) were risk factors found to be associated with H5N1 infection. In this study, we established an alternative serologic assay for H5N1 diagnosis, and we hereby present seroepidemiologic data of H5N1 infection in Northern Vietnam.

Molecular mechanism of inflammatory pain

Chronic inflammatory pain resulting from arthritis, nerve injury and tumor growth is a serious public health issue. One of the major challenges in chronic inflammatory pain research is to develop new pharmacologic treatments with long-term efficacy and few side effects. The mediators released from inflamed sites induce complex changes in peripheral and central processing by directly acting on transducer receptors located on primary sensory neurons to transmit pain signals or indirectly modulating pain signals by activating receptors coupled with G-proteins and second messengers. High local proton concentration(acidosis) is thought to be a decisive factor in inflammatory pain and other mediators such as prostaglandin, bradykinin, and serotonin enhance proton-induced pain. Proton-sensing ion channels [transient receptor potential V1(TRPV1) and the acid-sensing ion channel(ASIC) family] are major receptors for direct excitation of nociceptive sensory neurons in response to acidosis or inflammation.G-protein-coupled receptors activated by prostaglandin, bradykinin, serotonin, and proton modulate functions of TRPV1, ASICs or other ion channels, thus leading to inflammation- or acidosis-linked hyperalgesia. Although detailed mechanisms remain unsolved, clearly different types of pain or hyperalgesia could be due to complex interactions between a distinct subset of inflammatory mediator receptors expressed in a subset of nociceptors. This review describes new directions for the development of novel therapeutic treatments in pain.

Current challenges in diagnosis of lumbar radiculopathy

Lumbar radiculopathy(LR) is a term used to describe a pain syndrome caused by compression or irritation of nerve roots in the lower back. The surgery cost for LR increased by 23% annually during 1992-2003 in the developed country. Although it is one of most common complaints in clinical practice, the diagnosis for LR is still very challenging. Here we discuss the current tools of LR diagnosis and highlight the needs to develop new diagnosis tools for LR.

Autophagy in hepatitis C virus-host interactions:Potential roles and therapeutic targets for liver-associated diseases

Autophagy is a lysosome-associated,degradative process that catabolizes cytosolic components to recycle nutrients for further use and maintain cell homeostasis.Hepatitis C virus(HCV)is a major cause of chronic hepatitis,which often leads to end-stage liverassociated diseases and is a significant burden on worldwide public health.Emerging lines of evidence indicate that autophagy plays an important role in promoting the HCV life cycle in host cells.Moreover,the diverse impacts of autophagy on a variety of signaling pathways in HCV-infected cells suggest that the autophagic process is required for balancing HCVhost cell interactions and involved in the pathogenesis of HCV-related liver diseases.However,the detailed molecular mechanism underlying how HCV activates autophagy to benefit viral growth is still enigmatic.Additionally,how the autophagic response contributes to disease progression in HCV-infected cells remains largely unknown.Hence,in this review,we overview the interplay between autophagy and the HCV life cycle and propose possible mechanisms by which autophagy may promote the pathogenesis of HCVassociated chronic liver diseases.Moreover,we outline the related studies on how autophagy interplays with HCV replication and discuss the possible implications of autophagy and viral replication in the progression of HCV-induced liver diseases,e.g.,steatosis and hepatocellular carcinoma.Finally,we explore the potential therapeutics that target autophagy to cure HCV infection and its related liver diseases.

Target genes discovery through copy number alteration analysis in human hepatocellular carcinoma

High-throughput short-read sequencing of exomes and whole cancer genomes in multiple human hepatocellular carcinoma(HCC)cohorts confirmed previously identified frequently mutated somatic genes,such as TP53,CTNNB1 and AXIN1,and identified several novel genes with moderate mutation frequencies,including ARID1A,ARID2,MLL,MLL2,MLL3,MLL4,IRF2,ATM,CDKN2A,FGF19,PIK3CA,RPS6KA3,JAK1,KEAP1,NFE2L2,C16orf62,LEPR,RAC2,and IL6ST.Functional classification of these mutated genes suggested that alterations in pathways participating in chromatin remodeling,Wnt/β-catenin signaling,JAK/STAT signaling,and oxidative stress play critical roles in HCC tumorigenesis.Nevertheless,because there are few druggable genes used in HCC therapy,the identification of new therapeutic targets through integrated genomic approaches remains an important task.Because a large amount of HCC genomic data genotyped by high density single nucleotide polymorphism arrays is deposited in the public domain,copy number alteration(CNA)analyses of these arrays is a cost-effective way to reveal target genes through profiling of recurrent and overlapping amplicons,homozygous deletions and potentially unbalanced chromosomal translocations accumulated during HCC progression.Moreover,integration of CNAs with other high-throughput genomic data,such as aberrantly coding transcriptomes and non-coding gene expression in human HCC tissues and rodent HCC models,provides lines of evidence that can be used to facilitate the identification of novel HCC target genes with the potential of improving the survival of HCC patients.

Mechanisms inactivating the gene for E-cadherin in sporadic gastric carcinomas

AIM： To study the role of CDH1/E-cadherin （E-cad） gene alteration profiles including mutation, loss of heterozygosity （LOH）, promoter polymorphism and hypermethylation in mechanisms of CDH1 inactivation in gastric carcinoma （GC）. METHODS： Specimens were collected surgically from 70 patients with GC. Allelotyping PCR and detection of LOH, denaturing high pressure liquid chromatography and DNA sequencing, restriction fragment length polymorphism analysis, methylation specific PCR, and immunohistochemical staining were used. RESULTS： Promoter polymorphism was not a major mechanism of E-cad inactivation. Only one truncating mutation was found in a diffuse type tumor （3%）. Both LOH and promoter hypermethylation were major mechanisms of E-cad inactivation, but interestingly, there was a negative association between the fraction of allelic loss （LOH） in tumors and hypermethylation of CDH1. Therefore LOH and hypermethylation were two different tumorigenic pathways involved in GC. CONCLUSION： Given the findings that somatic mutation was extremely low and the relationship between LOH and hypermethylation was inverse, any two combinations of these three factors cannot fulfill the classical two-hit hypothesis of CDH1 inactivation. Thus, other mechanisms operating at the transcriptional level or at the post-translational level might be required to induce E-cadherin inactivation.

Helicobacter pylori infection in relation to E-cadherin gene promoter polymorphism and hypermethylation in sporadic gastric carcinomas

AIM: To study Helicobacter pylori (H pylori) infection in relation to E-cadherin (E-cad) promoter polymorphism and hypermethylation in GCs.METHODS: Specimens were taken from representative cancerous lesions and adjacent non-cancerous epithelia of 67 resected GCs. Hpyloriwas detected by real-time PCR of the cagA gene from non-neoplastic epithelium.E-cad promoter polymorphism and hypermethylation were determined by restriction fragment length polymorphism analysis and methylation-specific PCR, respectively. Expression of E-cad protein was determined by immunohistochemistry.RESULTS: Hpyloriwas found in 57% of patients with GC.H pylori infection was more frequently found in tumors with the -160C/C genotype than those with the -160C/A and -160A/A genotypes (74% vs47%, P = 0.02). Hpylori infection was associated with E-cad methylation in nonneoplastic epithelium; however, no significant difference in H pylori was observed between methylated and unmethylated cancerous lesions.CONCLUSION: Patients with the -160C/C genotype might require Hpyloriinfection to promote the inactivation of CDH1, suggesting that Hpylori infection might affect GC in an initial stage because polymorphism is germ line.Mechanism of hypermethylation of CDH1 promoter in GC is complex, and Hpyloriinfection might affect it in an initial stage.

Hepatitis B virus persistent infection-related single nucleotide polymorphisms in HLA regions are associated with viral load in hepatoma families

BACKGROUND Genome-wide association studies from Asia indicate that HLA-DP and HLA-DQ loci are important in persistent hepatitis B virus(HBV)infections.One of the key elements for HBV-related carcinogenesis is persistent viral replication and inflammation.AIM To examine genetic and nongenetic factors with persistent HBV infection and viral load in families with hepatocellular carcinoma(HCC).METHODS The HCC families included 301 hepatitis B surface antigen(HBsAg)carriers and 424 noncarriers born before the nationwide vaccination program was initiated in 1984.Five HBV-related single nucleotide polymorphisms(SNPs)—rs477515,rs9272105,rs9276370,rs7756516,and rs9277535—were genotyped.Factors associated with persistent HBV infection and viral load were analyzed by a generalized estimating equation.RESULTS In the first-stage persistent HBV study,all SNPs except rs9272105 were associated with persistent infection.A significantly higher area under the reciprocal operating characteristic curve for nongenetic factors vs genetic factors(P<0.001)suggests that the former play a major role in persistent HBV infection.In the second-stage viral load study,we added 8 HBsAg carriers born after 1984.The 309 HBsAg carriers were divided into low(n=162)and high viral load(n=147)groups with an HBV DNA cutoff of 105 cps/mL.Sex,relationship to the index case,rs477515,rs9272105,and rs7756516 were associated with viral load.Based on the receiver operating characteristic curve analysis,genetic and nongenetic factors affected viral load equally in the HCC family cohort(P=0.3117).CONCLUSION In these east Asian adults,the mechanism of persistent HBV infection-related SNPs was a prolonged viral replication phase.

Stem cell transplantation and/or adenoviral glial cell line-derived neurotrophic factor promote functional recovery in hemiparkinsonian rats

BACKGROUND Parkinson’s disease(PD)is a neurological disorder characterized by the progressive loss of midbrain dopamine(DA)neurons.Bone marrow mesenchymal stem cells(BMSCs)can differentiate into multiple cell types including neurons and glia.Transplantation of BMSCs is regarded as a potential approach for promoting neural regeneration.Glial cell line-derived neurotrophic factor(GDNF)can induce BMSC differentiation into neuron-like cells.This work evaluated the efficacy of nigral grafts of human BMSCs(hMSCs)and/or adenoviral(Ad)GDNF gene transfer in 6-hydroxydopamine(6-OHDA)-lesioned hemiparkinsonian rats.AIM To evaluate the efficacy of nigral grafts of hMSCs and/or Ad-GDNF gene transfer in 6-OHDA-lesioned hemiparkinsonian rats.METHODS We used immortalized hMSCs,which retain their potential for neuronal differentiation.hMSCs,preinduced hMSCs,or Ad-GDNF effectively enhanced neuronal connections in cultured neurons.In vivo,preinduced hMSCs and/or Ad-GDNF were injected into the substantia nigra(SN)after induction of a unilateral 6-OHDA lesion in the nigrostriatal pathway.RESULTS Hemiparkinsonian rats that received preinduced hMSC graft and/or Ad-GDNF showed significant recovery of apomorphine-induced rotational behavior and the number of nigral DA neurons.However,DA levels in the striatum were not restored by these therapeutic treatments.Grafted hMSCs might reconstitute a niche to support tissue repair rather than contribute to the generation of new neurons in the injured SN.CONCLUSION The results suggest that preinduced hMSC grafts exert a regenerative effect and may have the potential to improve clinical outcome.

Interactions between traffic air pollution and glutathione S-transferase genes on childhood asthma

AIM: To evaluate the role of glutathione S-transferase P1(GSTP1) genetic polymorphisms potentially modifying the association between NO2 and asthma/wheeze in Taiwan Residents children. METHODS: We investigated 3714 schoolchildren in Taiwan Children Health Study from 14 communities. Children's information was measured from questionnaire by parents. The traffic air pollutant was available from Environmental Protection Administration monitoring stations. RESULTS: A two-stage hierarchical model and a multiple logistic regression model were fitted to estimate the effects of NO2 exposures and GSTs polymorphisms on the prevalence of asthma and wheeze. Among children with GSTP1 Ile/Val or Val/Val genotypes, those residing in high-NO2 communities had significantly increased risks of asthma(OR = 1.76, 95%CI: 1.15-2.70), lateonset asthma(OR = 2.59, 95%CI: 1.24-5.41), active asthma(OR = 1.93, 95%CI: 1.05-3.57), asthma under medication(OR = 2.95, 95%CI: 1.37-6.32) and wheeze(OR = 1.54, 95%CI: 1.09-2.18) when compared with children in low-NO2 communities. Significant interactions were noted between ambient NO2 and GSTP1 on asthma, late-onset asthma, asthma under medication and wheeze(P for interaction < 0.05). However, we didnot find any association with polymorphisms in GSTM1 and GSTT1. CONCLUSION: Children under high traffic air pollution exposure are more susceptible to asthma, especially among those with GSTP1 Val allele.

Emerging roles of interferon-stimulated genes in the innate mmune response to hepatitis C virus infection

Infection with hepatitis C virus （HCV）, a major viral cause of chronic liver disease, frequently progresses to steatosis and cirrhosis, which can lead to hepatocellular carcinoma. HCV infection strongly induces host responses, such as the activation of the unfolded protein response, autophagy and the innate immune response. Upon HCV infection, the host induces the interferon （IFN）-mediated frontline defense to limit virus replication. Conversely, HCV employs diverse strategies to escape host innate immune surveillance. Type I IFN elicits its antiviral actions by inducing a wide array of IFN-stimulated genes （ISGs）. Nevertheless, the mechanisms by which these ISGs participate in IFN-mediated anti-HCV actions remain largely unknown. In this review, we first outline the signaling pathways known to be involved in the production of type I IFN and ISGs and the tactics that HCV uses to subvert innate immunity. Then, we summarize the effector mechanisms of scaffold ISGs known to modulate IFN function in HCV replication. We also highlight the potential functions of emerging ISGs, which were identified from genome-wide siRNA screens, in HCV replication. Finally, we discuss the functions of several cellular determinants critical for regulating host immunity in HCV replication. This review will provide a basis for understanding the complexity and functionality of the pleiotropic IFN system in HCV infection. Elucidation of the specificity and the mode of action of these emerging ISGs will also help to identify novel cellular targets against which effective HCV therapeutics can be developed.

Trap1a is an X-linked and cell-intrinsic regulator of thymocyte development

The X-linked Trap1a gene encodes the tumor rejection antigen P1A,which is expressed in fetal tissues and multiple lineages of tumor cells.The function of this gene remains unknown.Using chimeric mice with wild-type(WT)and Trap1a^(−/y)bone marrow,we show that Trap1a^(−/y)donor cells are capable of generating most lineages of hematopoietic cells,with the notable exception of T cells.Deletion of Trap1a selectively arrests T-cell development at double-negative stage 1(DN1,with a CD4^(−)CD8^(−)CD25^(−)CD44^(+)phenotype).Because Trap1a is expressed in Lin^(−)Sca-1^(+)c-Kit^(+)and common lymphoid progenitors but not in immature thymocytes(DN1-DN4),Trap1a mutations affect the differentiation potential of progenitor cells without directly acting on T cells.Despite a similarity in the blockade of DN1 to DN2 transition,the Trap1a^(−/y)DN1 cells have normal expression of c-Kit,in contrast to what was reported in the Notch1^(−/−)DN1.Complementary DNA profiling of Trap1a^(−/y)and WT embryonic stem cells shows that Trap1a does not regulate the Notch pathway.Our data reveal that Trap1a is an X-linked regulator that affects the differentiation potential of progenitor cells into T cells through a Notch-independent mechanism and identify an important function for the Trap1a gene.

‘Hide-then-hit’to explain the importance of genotypic polymorphism of DNA repair genes in determining susceptibility to cancer

have been suggested to account for different risk of developing cancers.In this review article,on the basis of breast cancer formation as a model,we propose a‘hide-then-hit’hypothesis indicating the importance of escaping checkpoint surveillance for sub-optimal DNA repair variants to cause cancer.Therefore,only cells with subtle defects in repair capacity arising from low-penetrance variants of DNA repair genes would have the opportunity to grow and accumulate the genetic changes needed for cancer formation,without triggering cell-cycle checkpoint surveillance.Furthermore,distinct from high-penetrance alleles,these polymorphic alleles of DNA repair genes would predispose carriers to a higher risk of developing cancer but would not necessarily cause cancer.To examine this,we simultaneously genotyped multiple SNPs of cell-cycle checkpoint genes and the DNA repair genes.Support for the hypothesis came from observations that breast cancer risk associated with variant genotypes of DNA repair genes became more significant in the subgroups of women with specific genotypic statuses of checkpoint genes.This‘hide-then-hit’hypothesis is certainly needed to be confirmed by biological evidence in which a cause–effect relationship has to be established.However,based on this,possible gene–gene interaction is considered to play an important role in modifying the cancer risk associated with genotypic polymorphism of DNA repair gene in different study populations.

The Study of IgG Subclass Profiles of Anti-HBc in Populations with Different Status of HBV Infection

To study IgG subclasses for the hepatitis B virus （HBV） core antigen （anti-HBc） in different populations, a comparison was made between 104 chronic carriers （60 male and 44 female） and 434 recovered individuals （247 male and 192 female）. Biochemistry analyses of AST （aspartate aminotransferase） and ALT （alanine aminotransferase） were also performed. Among the 104 chronic carriers, 21 patients were found to be ALT and AST abnormal （〉 25 IU/ml）. After comparing these ALT and AST abnormal patients with other ALT and AST normal chronic carriers, no statistical difference was observed in the OD values of the anti-HBe （p 〉 0.05）. The ELISA results showed the anti-HBc IgG subclass pattern was IgG1 〉 IgG3 〉 IgG4 in chronic carriers and IgG3 〉 IgG1 〉 IgG4 in recovered individuals （p 〈 0.05）. This result suggests the IgG1/IgG3 ratio may be related with HBV status. However, in spite of the different anti-HBc IgG1/IgG3 patterns demonstrated in different populations, both anti-HBc IgG1 and IgG3 concentrations were significantly higher in chronic carriers （p 〈 0.05）. Therefore, both the anti-HBc IgG1/IgG3 ratio and their amounts differed. They may play a significant role in chronic carriers and recovered individuals. The anti-HBc IgG subclass profiles of chronic carriers were not changed regardless of liver inflammation, and were independent of sex and age.