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.

Familial isolated pituitary adenoma (FIPA) is an autosomal dominant disease, characterized by low penetrance, early-onset disease, more invasive tumor growth, as well as somatotroph and lactotroph adenomas in most cases. It has been indicated that the aryl hydrocarbon receptor interacting protein (AIP) gene is a tumor suppressor gene. Many heterozygous mutations have been discovered in AIP in about 20% of FIPA families. However, the exact molecular mechanism by which its disfunction promotes tumorigenesis of pituitary is unclear.
Growth Hormone-Secreting Pituitary Adenoma ; genetics ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; Mutation ; Pituitary Neoplasms ; genetics

OBJECTIVETo investigate the effects of two fluid resuscitations on the bacterial translocation and the inflammatory factors of small intestine in rats with hemorrhagic shock.

METHODSFifty SD healthy male rats were randomly divided into 5 groups (n equal to 10 per group): Group A (Sham group), Group B (Ringer's solution for 1 h), Group C (Ringer's solution for 24 h), Group D (hydroxyethyl starch for 1 h) and Group E ((hydroxyethyl starch for 24 h). A model of rats with hemorrhagic shock was established. The bacterial translocation in liver, content of tumor necrosis factor-alpha (TNF-alpha) and changes of myeloperoxidase enzyme (MPO) activities in small intestine were pathologically investigated after these two fluid resuscitations, respectively.

RESULTSThe bacterial translocation and the expression of TNF-alpha in the small intestine were detected at 1 h and 24 h after fluid resuscitation. There were significant increase in the number of translocated bacteria, TNF-alpha and MPO activities in Group C compared with Group B, significant decrease in Group E compared with Group D and in Group B compared with Group D. The number of translocated bacteria and TNF-alpha expression significantly decreased in Group E as compared with Group C.

CONCLUSIONSThe bacterial translocation and the expression of TNF-alpha in the small intestine exist 24 h after fluid resuscitation. 6% hydroxyethyl starch can improve the intestinal mucosa barrier function better than the Ringer's solution.

Animals ; Bacterial Translocation ; drug effects ; Fluid Therapy ; Hydroxyethyl Starch Derivatives ; administration & dosage ; pharmacology ; Intestine, Small ; metabolism ; Isotonic Solutions ; administration & dosage ; pharmacology ; Male ; Peroxidase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Shock, Hemorrhagic ; therapy ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo investigate the relationship between abnormal ECG and pathologic changes in the cardiac conduction system (CCS).

METHODPathological changes of the CCS in 12 cases with abnormal ECG out of 16 pre-death ECG were observed.

RESULTS(1) Among 7 cases of sudden cardiac death, ECG monitoring recorded bradyarrhythmia in 6 cases, tachyarrhythmia 6 cases, bradycardia-tachycardia syndrome 2 cases, conduction block 6 cases, atrial premature beats 6 cases, ventricular premature beats 6 cases, and ST-T changes 4 cases. (2) The histopathological findings in the CCS were noted in all cases. Of these 12 cases, three had signs of fatty infiltration, and/or fibrous 4 cases, three of amyloidosis, one of chronic inflammatory changes, two of acute inflammatory changes, two of developmental anomalies, two of hemorrhages and one of LAD stenosis. (3) Acute inflammation changes in the CCS corresponded to tachyarrhythmia and multiple ventricular premature beats, whereas chronic inflammation and degenerative changes in the CCS were often related to bradyarrhythmia, bradycardia-tachycardia syndrome and conduction block. (4) The CCS changes alone could lead to ST-T changes in ECG.

CONCLUSIONThe pathological changes in the CCS are related to ECG changes, and attributed to the pathological bases of arrhythmia.

Adolescent ; Adult ; Aged ; Arrhythmias, Cardiac ; pathology ; physiopathology ; Child ; Electrocardiography ; Female ; Heart Conduction System ; pathology ; physiopathology ; Humans ; Male ; Middle Aged ; Young Adult

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

Adolescent ; Adult ; Ear Diseases ; congenital ; diagnosis ; surgery ; Female ; Fistula ; diagnosis ; surgery ; Humans ; Male ; Neck ; Young Adult

Objective To assess the efficacy and safety of mifepristone combined with oral or vaginal misoprostol for termination of pregnancy between 8 and 16 weeks of gestation. Methods This was a randomized, multi-center, open clinical trial. A total of 625 women at 8-16 weeks of gestation were randomized to receive 200 mg oral mifepristone followed by either oral misoprostol 400 μg every 3 hours or vaginal misoprostol 400μg every 6 hours for a maximum of 4 doses 36-48 hours later. There were 417 women in oral group with 198 at 8-9 weeks and 219 at 10-16 weeks, while 208 women in vaginal group with 99 at 8-9 weeks and 109 at 10-16 weeks. The outcome measures were the success abortion rate, induction to abortion interval, the amount of bleeding, reoccurrence of menstruation and adverse events. Results Abortion rate was significantly higher in vaginal group [98.1% (202/206)] than that in oral group [94.0%(390/415), P=0.023]; concerning termination of pregnancy at 8-9 weeks and 10-16 weeks respectively, there were no significant differences between oral and vaginal groups (P=0.156, P=0.073). The induction to abortion interval was no significant difference in oral and vaginal group in different gestational weeks ( P=0.238, P=0.273). The average induction to abortion interval was (4.1 ± 6.6) hours and (6.0 ± 4.5) hours respectively in terminating 8-9 weeks and 10-16 weeks of gestation. Concerning the amount of bleeding within 2 hours of placenta expulsion, there was significant difference between oral group [(63±46) ml] and vaginal group [(55 ± 45) ml] in terminating 8-9 weeks of gestation (P=0.047), while there was no significant difference between groups in terminating 10-16 weeks of gestation [oral group (76 ± 52) ml versus vaginal group (76 ± 61) ml, P=0.507]. The reoccurrence of menstruation was about 37 days in both oral and vaginal groups. Two cases of incomplete abortion were serious adverse events (SAE) relating to treatment. The common adverse events (AE) of nausea and vomiting were significantly higher in oral group [57.2% (239/417), 36.3% (151/417)] than those in vaginal group [45.4% (94/208), 26.1% (54/208); P=0.005, 0.011]. Conclusion Oral or vaginal misoprostol combined with mifepristone, is effective and safe for termination of pregnancy between 8 and 16 weeks of gestation.