Introduced in 2009, whole-exome sequencing (WES) is a technology in which target capture methods are used to enrich sequences of coding regions of genes from fragmented total genomic DNA, which is followed by high-throughput sequencing of the captured fragments. As reported, WES has been successfully applied for discovering genes underlying several Mendelian diseases, especially autosomal recessive types. In this review, authors have summarized the main computational strategies which have been applied to identify novel autosomal recessive diseases genes using whole-exome data.
Cloning, Molecular ; Exome ; Genes, Recessive ; Genetic Diseases, Inborn ; genetics ; Humans ; Sequence Analysis, DNA

OBJECTIVETo investigate the dynamic correlation between pre-S1 antigen, pre-S2 antigen and HBV DNA in the serum of chronic hepatitis B (CHB) patients undergoing nucleoside analogue therapy.

METHODS12 CHB patients with transient virological response after lamivudine treatment, and 20 patients treated with adefovir for 5 years were recruited in this study. Serum samples were collected at four time points when HBV DNA fluctuated sharply during lamivudine treatment, and at 0, 8, 12, 28, 52, 104, 156, 208, 260 weeks following adefovir treatment. HBV DNA was quantified by real-time PCR, pre-S1 and pre-S2 antigens were detected by ELISA.

RESULTSThe titers of pre-S1 and pre-S2 antigens were not correlated with the HBV DNA level in the serum of lamivudine treated patients. Only in one case of the adfovir treated patients, the decrease of pre-S1 and pre-S2 antigens was in parallel with the decrease of HBV DNA. Linear regression analysis indicated that neither pre-S1 antigen nor pre-S2 antigen was correlated with HBV DNA in the serum of lamivudine or adfovir treated patients (P more than 0.05).

CONCLUSIONOur results indicate that the titers of pre-S1 and pre-S2 antigens are not correlated with the serum HBV DNA in CHB patients undergoing nucleoside analogue therapy. Neither pre-S1 nor pre-S2 is a good predictor for the outcome of nucleoside analogue treatment.

DNA, Viral ; blood ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; drug therapy ; Humans ; Lamivudine ; therapeutic use ; Real-Time Polymerase Chain Reaction

OBJECTIVETo investigate the differential expressions of nucleolin in invasive cervical squamous cell carcinoma, cervical intraepithelial neoplasms (CIN) and normal cervical epithelial tissues and explore the role of nucleolin in the carcinogenesis and progression of cervical squamous cell carcinoma.

METHODSFifty specimens of invasive cervical squamous cell carcinoma, 65 specimens of CIN, and 60 adjacent normal cervical epithelial tissue specimens were examined immunohistochemically for nucleolin expression. The correlation of nucleolin expression levels with histological grades of invasive cervical squamous cell carcinoma and CIN were analyzed.

RESULTSThe specimens of invasive cervical squamous cell carcinoma showed a significantly higher positivity rate for nucleolin expression than CIN and normal cervical epithelial tissues, and the rate in CIN tissues was significantly higher than that in normal cervical epithelial tissues (P<0.01). The expression level of nucleolin was significantly higher in invasive cervical squamous cell carcinoma than in CIN and normal cervical epithelia tissues, and higher in CIN than in normal cervical epithelia tissues, whose immunostaining scores were 7.6±0.3, 6.1±0.2, and 3.0±0.2, respectively (P<0.01). The mean nucleolin immunostaining score was significantly higher in poorly and moderately differentiated than in highly differentiated cervical squamous cell carcinoma (7.9 vs 7.1, P<0.01), and higher in high grade CIN than in low grade CIN tissues (6.0 vs 4.0, P<0.01).

CONCLUSIONSOverexpression of nucleolin plays an important role during carcinogenesis of cervical squamous cell carcinoma and is positively correlated with tumor progression of CIN and cervical squamous cell carcinoma.

Carcinogenesis ; Carcinoma in Situ ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cervical Intraepithelial Neoplasia ; metabolism ; pathology ; Disease Progression ; Female ; Humans ; Phosphoproteins ; metabolism ; RNA-Binding Proteins ; metabolism ; Uterine Cervical Neoplasms ; metabolism ; pathology

OBJECTIVETo elucidate the characteristics of variation and the genetic evolution of non-structural protein (NS1, NS2) genes related to avian influenza subtype H5N1 viruses isolated from the boundary region of Yunnan province.

METHODSSwab samples were collected from foreign poultry and wild birds in the boundary regions of Yunnan province and screened by H5/N1 subtype-specific multiplex RT-PCR. The NS segment of H5N1 virus from the positive samples were amplified by RT-PCR and cloned into pMD18-T vectors for sequencing. The alignment and phylogenetic analysis on those available NS1, NS2 genes were performed with sequences of the known reference strains.

RESULTS71 positive samples were identified from 1240 samples, with the positive rate as 5.72%. Fourteen different NS segment sequences were obtained from 30 representative positive samples and could be divided into 3 distinct clades or sub-clades (I-1, I-2 and II), by phylogenetic analysis. The NS1/NS2 genes and Hemagglutinin (HA) genes of H5N1 viruses from the boundary regions of Yunnan province showed different relationships regarding the characteristics on genetic evolution. The substitution or mutation of key amino acids sites had been noticed in the nuclear location signal domains, effect domain, and other pathogenicity markers.

CONCLUSIONNS genes of H5N1 subtype viruses in boundary region of Yunnan province showed genetic divergence and the virus of clade I-2 and II had become dominant epidemic strains in this region since 2010.

Amino Acid Sequence ; Animals ; Animals, Wild ; Birds ; virology ; China ; epidemiology ; Evolution, Molecular ; Genome, Viral ; Influenza A Virus, H5N1 Subtype ; genetics ; isolation & purification ; Influenza in Birds ; epidemiology ; virology ; Phylogeny

OBJECTIVETo assess the clinic efficacy and safety in the treatment of chronic abacterial prostatitis/chronic pelvic pain syndrome (CABP/CPPS) after intraprostatic injection of Chuanshentong.

METHODSFive milliliter of solution blending Chuanshentong and lidocaine was transperineally injected into one lobe of prostate, once a day for 6 days, for a total of 98 cases of patients who had been diagnosed as chronic abacterial prostatitis/chronic pelvic pain syndrome. The efficacy was evaluated by the NIH Chronic Prostatitis Symptom Index (CPSI) after a 12 week follow-up.

RESULTSAll cases had completed the follow-up. Fifty-three cases (54.08%) were completely cured, accompanied by remarkable effective in 17 cases (17.35%) and improved in 23 cases (23.47%). The total remarkable efficacy was 71.43% and improved 94. 90%. Scores of CPSI decreased significantly compared with pre-treatment (95% confidence interval 12.85 approximately 17.91).

CONCLUSIONIt is suggested that transperineally intraprostatic injection of Chuanshentong may be a useful method for the treatment of CABP/CPPS.

Adult ; Chronic Disease ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Injections, Intralesional ; Male ; Middle Aged ; Pelvic Pain ; drug therapy ; Phytotherapy ; Prostatitis ; drug therapy

OBJECTIVEAlthough most of nerve injuries associated with Monteggia fracture-dislocation in children are neurapraxias and will recover spontaneously after conservative treatment, surgical exploration of the involved nerve is always required in the cases with the entrapment of posterior interosseous nerve (PIN). However, the necessity and time frame for surgical intervention for specific patterns of nerve dysfunction remains controversial. The aim of the report is to observe and understand the pathology of PIN injury associated with Monteggia fracture-dislocation in children, and to propose the possible indication for the exploration of nerve.

METHODSEight cases, six boys and two girls, with Monteggia fracture-dislocation complicated by PIN injury, managed operatively at the authors?Hospital from 2007 to 2008 were retrospectively reviewed. All the patients underwent the attempted closed reduction before they received exploration of PIN, with open reduction and internal fixation or successful closed reduction.

RESULTSThe PIN was found to be trapped acutely posterior to the radiocapitellar joint in 4 out of 5 Type III Bado's Monteggia fractures. In the remaining cases, since there were longer time intervals from injury to operation, chronic compressive changes and epineural fibrosis of radial nerve were visualized. After a microsurgical neurolysis performed, the complete recovery in the nerve function was obtained in all the cases during the follow-up.

CONCLUSIONThe findings from this study suggest that every case of type III Monteggia fracture-dislocation with decreased or absent function of muscles innervated by PIN and an irreducible radial head in children should be viewed as an indication for immediate surgical exploration of the involved nerve to exclude a potential PIN entrapment.

Female ; Fingers ; innervation ; Fracture Fixation, Internal ; Humans ; Male ; Monteggia's Fracture ; complications ; Muscle, Skeletal ; innervation ; Nerve Compression Syndromes ; etiology ; surgery ; Recovery of Function ; Retrospective Studies ; Thumb ; innervation ; Wrist ; innervation

BACKGROUNDPancreatic cancer is a highly malignant tumor affecting an ever increasing number of patients with a mean 5-year survival rate below 4%. Therefore, gene therapy for cancer has become a potential novel therapeutic modality. In this study we sought to determine the inhibitory effects of adenovirus-mediated human interleukin-24 (AdhIL-24) on pancreatic cancer.

METHODSHuman interleukin-24 gene was cloned into replication-defective adenovirus specific for patu8988 tumor cells by virus recombination technology. Reverse transcription-polymerase chain reaction and Western blotting analysis were used to determine the expression of human interleukin-24 mRNA in patu8988 cells in vitro. Induction of apoptosis by overexpression of human interleukin-24 in patu8988 cells was determined by flow cytometry. In vivo efficacy of adenoviral delivery of human interleukin-24 was assessed in nude mice (n = 10 for each group) bearing patu8988 pancreatic cancer cell lines by determining inhibition of tumor growth, endothelial growth factor and CD34 expression, and intratumoral microvessel density (MVD).

RESULTSThe recombinant adenovirus vector AdVGFP/IL-24 was constructed with a packaged recombinant retrovirus titer of 1.0 x 10(10) pfu/ml and successfully expressed of both mRNA and protein in patu8988 cells. The AdVGFP/IL-24 induced apoptosis of patu8988 tumor cells in vitro and significantly inhibited tumor growth in vivo (P < 0.05). The intratumoral MVD decreased significantly in the treated tumors (P < 0.05).

CONCLUSIONThe recombinant adenovirus AdGFP/IL-24 can effectively express biologically active human interleukin-24, which results in inhibition of pancreatic cancer growth.

Adenoviridae ; genetics ; Animals ; Antigens, CD34 ; analysis ; Blotting, Western ; Flow Cytometry ; Genetic Therapy ; Genetic Vectors ; Humans ; Interleukins ; genetics ; Male ; Mice ; Mice, Inbred BALB C ; Pancreatic Neoplasms ; pathology ; therapy ; Transfection ; Vascular Endothelial Growth Factor A ; analysis

OBJECTIVETo elucidate the variation in characterizations and genetic evolution of the matrix protein 2 or ion channel protein(M2) genes of avian influenza subtype H5N1 viruses in the boundary region of Yunnan province from 2008 to 2012.

METHODSA total of swab samples were collected from foreign poultry such as the junction between Yunnan and Vietnam, Laos,myanmar and wild birds in boundary region of Yunnan province from 2008 to 2012 and screened by H5N1 subtype-specific multiplex RT-PCR. The M genes of H5N1 virus from the positive samples were amplified by RT-PCR and cloned into pMD18-T vectors for sequencing. The alignment and phylogenetic analysis of M2 genes were performed with sequences of the known reference strains.

RESULTSA total of 71 positive samples were found out of 1240 samples and the positive rate was 5.72%. A total of 14 different M2 sequences were obtained from 30 positive samples and were divided into 3 distinct clades or sub-clades(1.2.1, 1.2.2 and 2) by phylogenetic analysis, 5, 7 and 2, respectively. The M2 genes and Hemagglutinin(HA) genes of H5N1 viruses from the boundary region of Yunnan province had showed different relationship of genetic evolution. The substitution or mutation of key amino acids sites had been found among the domains of epitope, adamantane-resistance, and poultry or human original viral strains.

CONCLUSIONThe M2 genes of H5N1 subtype viruses in boundary region of Yunnan province from 2008 to 2012 showed genetic divergence and the virus of clade 1.2.2 had become dominant epidemic strain in this region.

Animals ; Birds ; virology ; Chickens ; virology ; China ; Evolution, Molecular ; Influenza A Virus, H5N1 Subtype ; classification ; genetics ; Influenza in Birds ; virology ; Phylogeny ; Poultry ; virology ; Viral Matrix Proteins ; genetics

BACKGROUNDPatients with the genotypes of both CYP2C9*3/*3 and VKORC1-1639 A/A are expected to require the lowest dose of warfarin, and to have a greatly increased risk of bleeding. The experience for the dosing of warfarin in such extremely rare cases has been seldom reported.

METHODSDemographic and clinical data from two cases with stable low dose of warfarin in China were studied by resequencing the corresponding gene segments in their whole blood DNA. The potential clinical value of the pharmacogenetic algorithm for them was evaluated by calculating the stable dose of warfarin in pharmacogenetic algorithm developed by International Warfarin Pharmacogenetics Consortium.

RESULTSBoth cases (68-year-old female and 50-year-old male) were diagnosed as chronic nonvalvular atrial fibrillation needing warfarin treatment, with target international normalized ratio (INR) 2 to 3. Case 1 had stable warfarin dose of 0.625 mg/d and case 2 1.25 mg/d. They needed more than 1 month to stabilize their anticoagulation. Exceeding INR values were recorded for them when the dose of warfarin was no more than 2 mg/d. Hemorrhagic complication appeared in case 1 when the dose was titrated from 2.5 to 1.25 mg/d. No concomitant medicine to increase or decrease the INR value was recorded for them. Genotyping CYP2C9 and VKORC1 showed both patients were the carriers of the homozygous alleles -CYP2C9*3/*3 and VKORC1-1639 A/A. Their stable doses of warfarin calculated by the pharmacogenetic dose algorithm (0.672 mg/d for case 1 and 1.16 mg/d for case 2) were comparable with their actual stable therapeutic doses.

CONCLUSIONSTwo Chinese with the rare genotypes of both CYP2C9*3/*3 and VKORC1-1639 A/A were found to require the extremely low dose of warfarin. The pharmacogenetic algorithm incorporating the variances of VKORC1 and CYP2C9 genotypes, as well as the non-genetic factors could predict their stable dose of warfarin with high accuracy.

Aged ; Anticoagulants ; adverse effects ; Aryl Hydrocarbon Hydroxylases ; genetics ; Cytochrome P-450 CYP2C9 ; Female ; Genotype ; Hemorrhage ; chemically induced ; Humans ; Male ; Middle Aged ; Mixed Function Oxygenases ; genetics ; Pharmacogenetics ; Vitamin K Epoxide Reductases ; Warfarin ; adverse effects

The changes of microRNA expression in rat hippocampus after traumatic brain injury (TBI) were explored. Adult SD rats received a single controlled cortical impact injury, and the ipsilateral hippocampus was harvested for the subsequent microarray assay at three time points after TBI: 1st day, 3rd day and 5th day, respectively. We characterized the microRNA expression profile in rat hippocampus using the microRNA microarray analysis, and further verified microarray results of miR-142-3p and miR-221 using quantitative real-time PCR. Totally 205 microRNAs were identified and up-/down-regulated more than 1.5 times. There were significant changes in 17 microRNAs at all three time points post-TBI. The quantitative real-time PCR results of miR-142-3p and miR-221 indicated good consistency with the results of the microarray method. MicroRNAs altered at different time points post-TBI. MiR-142-3p and miR-221 may be used as potentially biological markers for TBI assessment in forensic practice.
Animals ; Biomarkers ; metabolism ; Brain Injuries ; metabolism ; pathology ; Female ; Forensic Genetics ; Gene Expression Profiling ; Gene Expression Regulation ; Hippocampus ; metabolism ; pathology ; Male ; MicroRNAs ; biosynthesis ; Rats ; Rats, Sprague-Dawley