Objective To set up a new diagnostic platform based on microarray exon-capture and next-generation sequencing for detecting small mutations in dystrophin gene.The sensitivity and specificity of the method were assessed in clinical settings and the distribution of small mutations in Chinese Duchenne muscular dystrophy/Becker muscular dystrophy (DMD/BMD) patients were also analyzed.Methods Forty-one DMD/BMD patients diagnosed by the clinical criteria without large deletion or duplication (≥ 1exon) were recruited from Peking Union Medical College Hospital consecutively.Genomic DNA was extracted from blood samples.The libraries were prepared.Then exon and intron-exon flanking sequences of DMD gene were captured by custom microarray.Targeted next-generation sequencing and Sanger Sequencing were conducted.The patients who were not detected any disease-causing mutation were performed muscle biopsy.Results Thirty-eight subjects were detected small mutations in DMD gene.All single nucleotide variants (SNVs) and insertion & deletions (INDELs) were validated by Sanger sequencing.Twenty-one novel mutations were reported.The distribution of SNVs and INDELs was similar to other international DMD databases.Upon immunohistochemistry staining of dystrophin protein,1 of 3 mutation-undetected patients was diagnosed as DMD,2 of them were excluded.The specificity of the method was 100％,while the sensitivity was 97.4％.Conclusions Our microarray-captured next-generation sequencing assay could detect SNVs and INDELs with high sensitivity and specificity.Its advantages are economic,time-saving and stable.The platform is suitable for clinical gene diagnosis.

Background and purpose: Primary signet ring cell carcinoma(SRCC) of the bladder is rarely diagnosed in the clinic. Few cases have been reported in the literature, so there was lack of understanding of the primary bladder SRCC in terms of diagnosis and treatment. Our study was to investigate the clinical features and treatment strategy for primary SRCC of the bladder and review the status of the disease along with the literature. Methods: 3 cases of primary bladder SRCC were studied, including clinical features, treatment, follow-up and their prognosis.The literature was reviewed. Results: All cases received ultrasound, computerized tomography, cystoscopy, biopsy and other related lab tests for diagnosis and differential diagnosis. Laparoscopic radical cystectomy and orthotopic ileal neobladders were performed in 2 cases, while the other case received laparoscopic radical cystectomy and ileal conduit diversion, Chemotherapy (cisplatin and 5-fluorouracil) was delivered in one case after surgery. One patient died at 6 months postoperatively because of multiple metastasis. The other 2 cases have been followed-up only for 8 and 12 months postoperatively, and no recurrence or metastasis have been observed. Conclusion: Primary SRCC of the bladder lacks distinctive clinical and imaging manifestations. The tumor grows very invasively. Radical cystcctomy is one of the optimal approaches for treatment of SRCC of bladder.

Background and purpose: The prognostic factors on survival for the patients with prostate carcinoma are still underdeterrnined. This study was to analyze the survival of three common treatment methods for prostate carcinoma and the prognostic factors on survival. Methods: 494 male patients who were diagnosed as prostate cancer were enrolled into the retrospective study. All of the data like age, stage, grade, PSA level, ALP, Hb and treatments were collected. Overall survival and disease specific survival rates for patients were analyzed by Kaplan-Meier method. Prognostic factors on disease specific survival were also analyzed by Log-rank test and Cox proportional hazards model. Results: Disease specific survival rates at 1, 3 and 5 year were 96.0%, 89.0% and 80.0% for all 494 patients, respectively. Disease specific survival rate at 3-year was 92.4% for brachytherapy, 100.0% for radical prostatectomy and 80.6% for hormonal therapy (P=0.008). Multivariate analysis by Cox model showed that stage, PSA level and age significantly impacted on disease specific survival. Conclusion: Brachytherapy and radical prostatectomy provides longer survival time than hormonal therapy for patients with prostate cancer. Clinical stage and PSA level and age of prostate cancer are independent factors impacting on survival significantly.

Objective To investigate the expression of cathepsin B (CB) in human gastric carcinoma tissue.Methods The expression of CB in human gastric tissue was studied by using monospecific polyclonal rabbit antibody raised against human liver CB for immunohistochemistry, and full length cDNA of CB for in situ hybridization and dot blot.Results CB overexpression in gastric carcinoma was found when compared with non-neoplastic gastric tissue at both mRNA and protein levels. Diffuse cytoplasmic CB staining of mRNA and protein were identified in malignant cells of 53.3% and 69.1% of gastric adenocarcinoma respectively. The increased staining of CB in malignant cells was associated with the depth of the invasiveness and growth pattern as well as metastasis of lymph nodes, but not with the histological classification. It was also found that there were the expression of CB in stromal cells of the tumor and the expression localized mainly in the endothelial cells of the microvessels which correlated with angiogenesis. Conclusion These results indicate that the expression of CB in gastric carcinoma is related to tumor progression, and leads to development of the invasive phenotype.

UNLABELLEDTo screen and analyze the apoptosis- and proliferation-related genes in human nasopharyngeal carcinoma (NPC).

METHODSAccording to gene ontology classification, the abnormal expressions of the genes related to cell apoptosis and proliferation were identified in the NPC gene chip data. The cell apoptosis- and proliferation-related genes expressed in each of the 3 stages, as defined by the tree model for the pathogenesis and progression of NPC, were screened, and with literature review, their distribution in the tree model were analyzed.

RESULTSNineteen genes related to cell apoptosis were found in NPC, among which 9 were down-regulated (such as DNASE1L3) and located in the chromosome deletion regions, and 10 were up-regulated (such as DEDD) in the chromosome amplification regions. Twenty-one cell proliferation-related genes were identified, including 8 down-regulated genes (such as TUSC2) in the chromosome deletion regions and 13 up-regulated ones (such as EMP1) in the chromosome amplification regions. In the chromosome deletion regions, the down-regulated cell apoptosis-related genes participated mostly in inducing and regulating cell apoptosis, and the up-regulated cell proliferation-related genes in the chromosome amplification regions were mostly associated with the positive regulation of cell proliferation.

CONCLUSIONNPC occurs possibly through two pathways by inhibiting cell apoptosis or by promoting excessive cell proliferation.

Apoptosis ; genetics ; Cell Proliferation ; Chromosome Deletion ; Down-Regulation ; Gene Expression Profiling ; Humans ; Nasopharyngeal Neoplasms ; genetics ; pathology ; Oligonucleotide Array Sequence Analysis ; Up-Regulation

OBJECTIVEThe purpose was to analyze the effects of three sterilization methods (dry heat sterilization, steam sterilization, and chemical sterilization) on the corrosion of dental fissure bur.

METHODS200 dental fissure burs were distributed to 10 groups. Bending strength, elastic modulus, and torsional strength were measured by bending and torsional instrument and calculated with special designed software. Among the three sterilization methods, the steam sterilization group showed the most evident.

RESULTSThe corrosion was most severe in steam sterilization group, followed by chemical sterilization, dry heat sterilization. With the sterilization time increased, bending strength, elastic modulus, and torsional strength decreased respectively. Of the three sterilization methods, the mechanical properties were decreased most evidently by steam sterilization, followed by chemical sterilization and dry heat sterilization.

CONCLUSIONIt is proved that the bending strength, elastic modulus and torsional strength have a tight relationship with the corrosion of dental fissure burs. The corrosion was most severe in steam sterilization group, followed by chemical sterilization, dry heat sterilization. In regards of the corrosive effect, the dry heat sterilization might be the best way to sterilize the dental fissure burs.

Dental Fissures ; Dental High-Speed Equipment ; Dental Instruments ; Steam ; Sterilization

OBJECTIVETo explore whether bone marrow stem cells (MSCs) from adult mice can be induced to differentiate into hepatocytes by hepatocyte growth factor (HGF) alone and the time phase characteristics in the differentiation progress.

METHODSAdult mouse MSCs were treated with or without 100 ng/ml HGF, on days 0, 7, 14, 21, and 28. The morphologic characteristics of the cells were examined; the albumin (ALB), AFP mRNA was analyzed sub-quantively using reverse transcription polymerase chain reaction (RT-PCR) and immumohistochemistry techniques. The expression of ALB, AFP and CK19 were detected by using anti-ALB, AFP and CK19 antibodies.

RESULTSFreshly isolated adult mouse MSCs expressed ALB and AFP mRNA weakly; in the group without HGF, no ALB mRNA was detected on day 7. The expression of AFP mRNA was reduced significantly on day 7, and could not be detected anymore after day 14. In the HGF treated group, ALB mRNA was not detected on day 7, but the positive lane appeared again on day 14, and the expression of ALB mRNA was increased on day 21 but reduced in the following days. The AFP mRNA was positive at all times, however it tended to decrease after day 14 in the HGF treated groups. The result of immumohistochemistry was consistent with that of RT-PCR, and CK19 was always negative.

CONCLUSIONAdult mouse MSCs can be induced into hepatocyte differentiation in vitro. The optimal time for the induction was 2 to 3 weeks.

Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Hepatocyte Growth Factor ; pharmacology ; Hepatocytes ; cytology ; Male ; Mice ; Stem Cells ; cytology ; Time Factors

Computer Graphics ; Computer-Assisted Instruction ; Forensic Pathology ; Humans ; Imaging, Three-Dimensional ; Pathology ; education ; Pathology, Clinical ; Specimen Handling ; methods

OBJECTIVETo evaluate the therapeutic effect of laser photocoagulation of ciliary processes after pars plana vitrectomy in aphakic glaucoma.

METHODSTwenty patients (20 eyes) of aphakic glaucoma underwent vitrectomy plus laser treatment. During the surgery, after conventional 3-incision pars plana vitrectomy, a probe of semi-conductor diode laser (532 nm) was inserted through the sclera incision, the ciliary processes were then photocoagulated under direct visualization for 180 degree range. Before and after the surgery, the visual acuity, the intraocular pressure (IOP) as well as the outer appearance of the anterior segment were evaluated. The mean follow-up period was 13 months.

RESULTSThe IOP at the last visit was (21.35 +/-2.52) mmHg, which was significantly lower than that before the surgery [(39.75 +/-6.27) mmHg, P=0.000]. Atrophy of the ciliary processes was observed 1-3 months after the surgery.

CONCLUSIONPars plana vitrectomy combined with laser coagulation of ciliary processes reduces the IOP in patients with aphakic glaucoma effectively.

Adult ; Aphakia ; complications ; Ciliary Body ; surgery ; Female ; Glaucoma ; complications ; physiopathology ; surgery ; Humans ; Intraocular Pressure ; Laser Coagulation ; methods ; Male ; Middle Aged ; Treatment Outcome ; Visual Acuity ; Vitrectomy ; methods

OBJECTIVETo characterize the role of Toll-like receptor 4 (TLR4) in silica-induced production of tumor necrosis factor alpha (TNFalpha) from macrophage cell line.

METHODSThe human macrophage cell line THP-1 was incubated with silica suspension. Cell media were collected and TNFalpha levels in the supernatants measured with ELISA. To examine the involvement of TLR4 in silica-induced TNFalpha release, the neutralizing antibody (HTA125) against human TLR4 receptor was employed to pretreat THP-1 cells prior to silica treatment. Moreover, murine macrophages expressing wild type or mutated TLR4 were also treated with silica to verify the effect of TLR4 in silica-induced TNFalpha release.

RESULTSCompared with the control group [(3.18 +/- 0.41) pg/ml], the TNFalpha release in cells exposed to 100 microg/ml silica for 4 h and 8 h [(4.71 +/- 0.84), (6.22 +/- 0.58) pg/ml, respectively] increased 1.48 and 1.96 fold, respectively. Pretreatment of THP-1 cells with 20 microg/ml HTA125 antibody significantly blocked silica-induced TNFalpha release by 27%. Furthermore, the TNFalpha content released from cells expressing mutated TLR4 reduced by 30% in compared with that from the cells expressing wild type TLR4 after silica stimulation.

CONCLUSIONTLR4 mediates silica-induced TNFalpha release from macrophages.

Antibodies ; pharmacology ; Cell Line ; Humans ; Macrophages ; drug effects ; metabolism ; Silicon Dioxide ; toxicity ; Toll-Like Receptor 4 ; immunology ; Tumor Necrosis Factor-alpha ; metabolism