BACKGROUNDThymokidney has been reported as an approach for a vascularized thymus for transplantation to induce donor specific tolerance. A completely thymectomized model which ensures that the obtained thymus is not injured has not been developed yet and it would be useful for evaluating autologous thymokidney function in rats.

METHODSAdult Sprague-Dawley male rats weighing 150 - 300 g (n = 30) underwent non-invasive intubation with the assistance of an improved self-made wedge-shaped cannula made from a 2-ml plastic syringe and transillumination from the anterior tracheal area by an operation spotlight. The rats then received a thoracotomy while their breathing was supported by a small animal ventilator, and both lobes of the thymus were entirely extirpated under a 10× microscope. The postoperative survival rate of the rats was recorded, and changes in the T-cell reservoir from 9 of 30 rats within 21 days after surgery were monitored using flow cytometry. The complete thymectomy rate was confirmed by autopsy and histological examination on 21 days post-operation.

RESULTSThe postoperative survival rate of rats was 100%. The exsected thymus was free of injury and the rate of complete thymectomy was 100%.

CONCLUSIONSThis model has a stable survival rate and complete thymectomy is able to be achieved. The obtained thymus tissue is free of injury and can be used for transplantation.

Animals ; Intubation, Intratracheal ; methods ; Male ; Rats ; Rats, Sprague-Dawley ; Thoracotomy ; methods ; Thymectomy ; methods ; Thymus Gland ; surgery

OBJECTIVETo study the transmission route and epidemiological features of Clonorchis sinensis infection in Shenzhen area--the biggest immigration city of Southern China.

METHODSIn this study, we examined 1473 individuals (710 males and 763 females) to determine the current status of C. sinensis infection among the people in one village in Zhujiang river region, Guangdong province, China. Blood samples were detected on antibody of C. sinensis with enzyme linked immunosorbent assay,and stool specimens from sera positive cases were examined by modified Kato-Katz thick smear to confirm the density of infection. People were interviewed on their life styles under the structured questionnaire which was administered by trained staff members. Major content of the questionnaire included eating raw fish, using the same utensils for both raw fish and cooked food, using feces of domestic animals and human feces to feed fish and so on.

RESULTSAmong 1473 people examined, 70 (4.75%) were found infected with C. sinensis. By counting eggs per gram feces (EPG), it was found that heavy intensities of infection in males was stronger than that of females,and the overall average EPG was 41.87. Of 1473 interviewees, 54% of them did not know about fluke disease or its transmission route, 12% of those who knew about the fluke but believed that the infection caused no harm or only slight harm to their health. 27% of the interviewees ate raw fish at least 1-2 times per months with 5% of the families using the same utensils for both raw fish and cooked food. 40% of the fish ponds owners fed their fish with the feces of domestic animals and human feces.

CONCLUSIONTogether with these results, unhealthy behaviors, poor knowledge, inappropriate farming/fishery practices, eating raw fish were important factors influencing the C. sinensis prevalence in humans.

Adolescent ; Adult ; Animals ; Child ; China ; epidemiology ; Clonorchiasis ; epidemiology ; parasitology ; transmission ; Clonorchis sinensis ; pathogenicity ; Feces ; parasitology ; Female ; Humans ; Male ; Middle Aged ; Parasite Egg Count ; Reverse Transcriptase Polymerase Chain Reaction ; Sex Factors ; Surveys and Questionnaires ; Young Adult

OBJECTIVE: To study the suspected autosomal STR loci mutation cases. METHODS: A total of 227 suspected autosomal STR loci mutation cases were selected from Center of Forensic Sciences, Beijing Genomics Institute. The allelic mutation cases were screened and the number of mutation of each STR loci was statistically analyzed. The CPI value was calculated in order to study the characteristics and rules of the mutations. RESULTS: In the 227 suspected mutation cases, 3 cases were excluded paternity, and 228 mutations were observed at 18 STR loci in the rest of the cases. The average number of STR mutation loci was 1-2. The maximum of mutation step was 4. After using 20A amplification kit, the CPI values in 3 non-parentage cases were all less than 10(4). After using 20A and 10G amplification kits, the CPI values were all larger than 10(4) in all standard parents-child triplet cases and in 99.45% of diad cases. CONCLUSION The allelic mutation of STR loci is relatively common in forensic cases. By increasing the number of the required STR loci and supplementing the samples of the triplet, the identification errors could be decreased to a great extent when suspected autosomal STR loci mutation occurs.
DNA Fingerprinting ; Female ; Forensic Medicine ; Gene Frequency ; Genetic Loci ; Humans ; Male ; Microsatellite Repeats ; Mutation ; Paternity ; Reagent Kits, Diagnostic ; Retrospective Studies

OBJECTIVETo explore the role of sorafenib in reversing multidrug resistance (MDR) in hepatoma BEL-7402/FU cells and its possible mechanisms.

METHODSMTT colorimetric assay was used to obtain the dose-response curve of sorafenib in BEL-7402/FU cells, and flow cytometry performed to assess the effect of sorafenib on Rho123 concentration in the cells. The optimal dose of sorafenib for cell treatment was determined according to the results of MTT assay and flow cytometry. MTT assay was employed to evaluate the effect of sorfenib on the cytotoxicity of the antitumor drugs, flow cytometry performed to determine the expression of cell membrane transport protein (P-gp), and RT-PCR used to detect mdr1 gene expression in the cells treated with sorafenib at the optimal dose.

RESULTSSorafenib at the concentration of 4 micromol/L, efficiently reversed the MDR of the cells with minimal side effects. At the concentration of 4 micromol/L, sorafenib partially reversed the drug resistance of BEL-7402/FU cells to ADM, 5-FU, GEM and DDP, with reversal indexes of 2.98, 7.16, 1.99 and 10.08, respectively. Treatment of the cells with 4 micromol/L, sorafenib also partially down-regulated P-gp expression in BEL-7402/FU cells, and caused a reduction of mdr1 gene expression by 27.3% in comparison with the control cells.

CONCLUSIONSorafenib can reverse MDR in human hepatoma cells probably in association with down-regulation of mdr1 gene expression and increased accumulation of the chemotherapeutic agents in the cells.

ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Antineoplastic Agents ; pharmacology ; Benzenesulfonates ; pharmacology ; Cell Line, Tumor ; Down-Regulation ; drug effects ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Humans ; Liver Neoplasms ; genetics ; Niacinamide ; analogs & derivatives ; Phenylurea Compounds ; Pyridines ; pharmacology

OBJECTIVETo construct a database of human lung squamous carcinoma cell line NCI-H226 and to facilitate discovery of novel subtypes markers of lung cancer.

METHODProteomic technique was used to analyze human lung squamous carcinoma cell line NCI-H226. The proteins of the NCI-H226 cells were separated by two-dimensional gel electrophoresis and identified by mass spectrometry.

RESULTSThe results showed that a good reproducibility of the 2-D gel pattern was attained. The position deviation of matched spots among three 2-D gels was 1.95 +/- 0.53 mm in the isoelectric focusing direction, and 1.73 +/- 0.45 mm in the sodium dodecyl sulfate-polyacrylamide gel electrophoresis direction. One hundred and twenty-seven proteins, including enzymes, signal transduction proteins, structure proteins, transport proteins, etc. were characterized, of which, 29 identified proteins in NCI-H226 cells were reported for the first time to be involved in lung cancer carcinogenesis.

CONCLUSIONThe information obtained from this study could provide some valuable clues for further study on the carcinogenetic mechanism of different types of lung cancer, and may help us to discover some potential subtype-specific biomarkers of lung cancer.

Biomarkers, Tumor ; analysis ; Carcinoma, Squamous Cell ; chemistry ; Cell Line, Tumor ; Databases, Factual ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Lung Neoplasms ; chemistry ; Neoplasm Proteins ; analysis ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Tandem Mass Spectrometry

OBJECTIVETo investigate the role of Th17 cells in immune thrombocytopenia (ITP) mice model.

METHODITP was induced by daily intraperitoneal injection of anti-platelet membrane CD41 antibody (MWReg30) into BALB/c mice, the mRNA expressions of Th17 cell associated transcription factors and cytokines in peripheral blood and spleen mononuclear cells were measured by real-time PCR, and the proportion of Th17 cells by FCM analysis.

RESULTSThe percentage of Th17 cell was significantly elevated in ITP mice both in splenocyte and peripheral blood as compared with that in normal controls (P<0.01). ITP mice had elevated mRNA expressions of IL-17F, IL-17A and IL-6 in splenocyte (P<0.05), and of IL-21 in peripheral blood (P<0.05). There was a positive correlation between IL-17A and IL-17F (r = 0.934, P = 0.000), and between IL-17A/IL-17F and IL-6 (r = 0.598, P = 0.001; r = 0. 619, P = 0.000).

CONCLUSIONSTh17 cell might play an important role in the pathogenesis of ITP, at least involving in the clearance of platelets.

Animals ; Disease Models, Animal ; Female ; Mice ; Mice, Inbred BALB C ; Th17 Cells ; immunology ; Thrombocytopenia ; immunology

OBJECTIVETo review the progress of cardiac differentiation and electrophysiological characteristics of bone marrow mesenchymal stem cells.

DATA SOURCESThe databases of PubMed, Springer Link, Science Direct and CNKI were retrieved for papers published from January 2000 to January 2012 with the key words of "bone marrow mesenchymal stem cells, cardiac or heart, electrophysiology or electrophysiological characteristics".

STUDY SELECTIONThe articles concerned cardiac differentiation and electrophysiological characteristics of bone marrow mesenchymal stem cells were collected. After excluding papers that study purposes are not coincident with this review or contents duplicated, 56 papers were internalized at last.

RESULTSFor the treatment of myocardial infarction and myocardiac disease, the therapeutic effects of transplantation of bone marrow mesenchymal stem cells which have the ability to develop into functional myocardial cells by lots of methods have been proved by many researches. But the arrhythmogenic effect on ventricles after transplantation of bone marrow mesenchymal stem cells derived myocardial cells is still controversial in animal models. Certainly, the low differentiation efficiency and heterogeneous development of electrical function could be the most important risk for proarrhythmia.

CONCLUSIONMany studies of cardiac differentiation of bone marrow mesenchymal stem cells have paid attention to improve the cardiac differentiation rate, and the electrophysiology characteristics of the differentiated cells should be concerned for the risk for proarrhythmia as well.

Bone Marrow Cells ; cytology ; Cell Differentiation ; physiology ; Electrophysiology ; Humans ; Mesenchymal Stromal Cells ; cytology ; Myocardial Infarction ; therapy ; Myocytes, Cardiac ; cytology ; physiology

This study investigated the role of 15-hydroxyeicosatetraenoic acid (15-HETE) in rabbit pulmonary arterial smooth muscle cells (PASMCs) under hypoxia by using organ bath and whole cell patch-clamp techniques. Neonatal rabbits born into normoxic environment were transferred after first feeding into normal and hypoxic environments with respectively 0.21 and 0.12 fractional inspired oxygen (FiO2). Pulmonary arteries were extracted after 9 d and cut into rings 1.0 approximately 1.5 mm in length for organ bath experiments. Whole cell patch-clamp technique was used to measure the potassium current in the freshly dispersed rabbit PASMCs. The results showed that 15-HETE-induced vasoconstriction was blocked by 4-aminopyridine (5 mmol/L), a Kv channel blocker. The K(ATP) channel blocker glyburide (1 micromol/L) and the BKCa channel blocker tetraethylammonium (10 mmol/L) did not abolish this vasoconstriction. 15-HETE decreased the whole-cell voltage-gated K+ current in the PASMCs. These findings demonstrate that hypoxia blocks Kv channels through a 15-HETE mediated mechanism, leading to PA vasoconstriction.
Animals ; Animals, Newborn ; Cell Hypoxia ; Cells, Cultured ; Female ; Hydroxyeicosatetraenoic Acids ; pharmacology ; Muscle Contraction ; drug effects ; Muscle, Smooth, Vascular ; cytology ; metabolism ; physiology ; Potassium Channel Blockers ; pharmacology ; Potassium Channels, Voltage-Gated ; drug effects ; Pregnancy ; Pulmonary Artery ; cytology ; metabolism ; physiology ; Rabbits ; Vasoconstrictor Agents ; pharmacology

Objective To analyze the factors that might affect the recurrence after endoscopic resection of colorectal adenoma,we aim to provide a reference for the regular follow-up of colorectal adenoma after endoscopic resection.Methods In the Department of Gastroenterology,Huadong Hospital from Jan.,2013 to Dec.,2014,patients received endoscopic resection during hospital stay and postoperative follow-up colonoscopy.We collected their information of gender,age,smoking history,drinking history,hypertension,diabetes,fatty liver,cholecystectomy history,Helicobacter pylori (H.pylori) infection,related metabolic indicators,number of adenoma,size of adenoma,location of adenoma and pathology of adenoma.Multivariate and univariate analysis were performed by Logistic regression analysis to explore factors influencing the recurrence risk of colorectal adenoma.Results A total of 283 cases were enrolled.The recurrence rate of low-risk and high-risk adenoma was 39.3 ％ and 56.3 ％,respectively,and therate of all adenoma was 52.7％.Multivariate and univariate analysis both found that H.pylori infection (P＜0.001,OR:3.316 and 2.802,95％CI:1.869-5.884 and 1.660-4.728) and adenoma number (P＜0.001,OR:2.799 and 2.789,95％ CI:1.578-4.963 and 1.667-4.668) were risk factors for colorectal adenoma recurrence after endoscopic excision.Conclusions H.pylori infection and the number of base colorectal adenoma maybe associated with colorectal adenoma recurrence.

Objective To detect the rate of Angiostrongylus cantonensis infection and to study the effects of treatment so as to prepare monoclonal antibodies (McAbs) ,and gold immunochroma-tography assay (GICA) with 12D5 and 21B7 McAbs could be prepared in advance. Methods Two McAbs (12D5 and 21B7) were applied to detect the circulating antigen (CAg) in the sera of rats infected with A. cantonensis and angiostrongyliasis patients respectively by double antibody sandwich ELISA. Either 12D5 or 21B7 McAbs was used as antibody and protein A was conjugated with colloid gold as the detection marker. A special pad for GICA was designed according to the reaction procedure, and CAg were detected by GICA in the sera of rats infected with A. cantonensis and angiostrongyliasis patients respectively. Results 12D5 McAb was identified as IgG1 and 21B7 McAb was IgM. Results from Western blotting showed that two McAbs could be used to identified 55 KD protein of adult worms of A. cantonensis. The detection rates of CAg in the sera of infected rats was 100% (48/48) and the detection rates of CAg in the sera of angiostrongyliasis patients was 100% (32/32). No cross-reaction to sera of patients with other infection of parasites, such as clonochiasis, fasiolopsiasis, ancylostotniasis, ancylostomiasis, anisakiasis as well as schsitosomiasis wee seen and normal sera did not react with 12D5 and 21B7 McAbs. Conclusion Results from sandwich ELISA and GICA with 12D5 and 21B7 McAbs showed high specificity and acting as detecting CAg of A. cantonensis in sera of infected animals and patients. We noticed that GICA with 12D5 and 21B7 was not only rapid and simple that without requirement of special instrument, but also rather sensitive and specific for the detection of current infection with A.cantonensis.