Aim To Compare the quality of Pinellia Tubers between Nanchong Sichuan and relevant areas in order to standardize plant and to provide evidence for further exploitation and application. Methods Expectorant experiment was conducted with methods of washing mice's respiratory tract with phenol red and antitussive effect, with ammomia caused mice cough. Results In the experiment of expectorant effect, oral administration of the water or the ethanol extracts of pinellia Tubers showed no obvious expectorant effect, it also showed no significant difference compared with control NS. In the antitussive effect experiment, however the water or the ethanol extracts of pinellia Tuhers prolonged the incubation period and reduced the frequency of coughing. It presented significant difference compared with the control NS and clear time-effect relationship. Conclusion Pinellia Tubers from Nanchong Sichuan and relevant areas showed no apparent expectorant effect and have destinct antitussive effect. As for antitussive effects, water extracts were stronger than those of ethanol extracts, and wild Pinellia Tubers outmatched planted ones. Antitussive effect of wild Pinellia Tubers samples was rather stronger from Nanchong, Guangan, Suining, and Liangshan of Sichuan and Xihe Gansu.

Objective To investigate the clinical effect of Levosimendan and Qiliqiangxin Capsule in congestive heart failure and influence in serum levels of NT-proBNP and Hcy.Methods 92 cases of patients with congestive heart failure in our hospital from January 2014 to June 2016 were selected and divided into observation group and control group,46 cases in each group.Patients in the control group were treated with Levosimendan on the basis of conventional treatment,and the observation group were treated with Levosimendan and Qiliqiangxin Capsule.Compared the clinical effect,safety,and the change of serum levels of NT-proBNP and Hey before and after treatment.Results The total effective rate of observation group were higher than control group,but the difference was no significant.After treatment,the left ventricular ejection fraction (LVEF),stroke volume (SV) in two groups were significantly higher than that before treatment,the left ventricular end diastolic diameter (LVEDD) were significantly lower than before treatment (P ＜ 0.05);and the LVEF and SV of the observation group were significantly higher than the control group,LVEDD was significantly lower than the control group (P ＜ 0.05).After treatment,serum levels of NT-proBNP and Hey of two groups were significantly better than before (P ＜ 0.05),and the observation group were better than control group (P ＜ 0.05).The difference in the adverse reaction rate was no significant.Conclusion The clinical curative effect of Levosimendan and Qiliqiangxin Capsule in congestive heart failure is distinct,which can effectively improve the clinical symptoms of patients,the cardiac function,serum levels of NT-proBNP and Hcy.

BACKGROUNDGlioblastoma is the most common and lethal cancer of the central nervous system. Global genomic hypomethylation and some CpG island hypermethylation are common hallmarks of these malignancies, but the effects of these methylation abnormalities on glioblastomas are still largely unclear. Methylation of the O6-methylguanine-DNA methyltransferase promoter is currently an only confirmed molecular predictor of better outcome in temozolomide treatment. To better understand the relationship between CpG island methylation status and patient outcome, this study launched DNA methylation profiles for thirty-three primary glioblastomas (pGBMs) and nine secondary glioblastomas (sGBMs) with the expectation to identify valuable prognostic and therapeutic targets.

METHODSWe evaluated the methylation status of testis derived transcript (TES) gene promoter by microarray analysis of glioblastomas and the prognostic value for TES methylation in the clinical outcome of pGBM patients. Significance analysis of microarrays was used for genes significantly differently methylated between 33 pGBM and nine sGBM. Survival curves were calculated according to the Kaplan-Meier method, and differences between curves were assessed using the log-rank test. Then, we treated glioblastoma cell lines (U87 and U251) with 5-aza-2-deoxycytidines (5-aza-dC) and detected cell biological behaviors.

RESULTSMicroarray data analysis identified TES promoter was hypermethylated in pGBMs compared with sGBMs (P < 0.05). Survival curves from the Kaplan-Meier method analysis revealed that the patients with TES hypermethylation had a short overall survival (P < 0.05). This abnormality is also confirmed in glioblastoma cell lines (U87 and U251). Treating these cells with 5-aza-dC released TES protein expression resulted in significant inhibition of cell growth (P = 0.013).

CONCLUSIONSHypermethylation of TES gene promoter highly correlated with worse outcome in pGBM patients. TES might represent a valuable prognostic marker for glioblastoma.

Azacitidine ; analogs & derivatives ; pharmacology ; Brain Neoplasms ; drug therapy ; genetics ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cytoskeletal Proteins ; genetics ; DNA Methylation ; Glioblastoma ; drug therapy ; genetics ; pathology ; Humans ; LIM Domain Proteins ; genetics ; Promoter Regions, Genetic ; Treatment Outcome

An about 700 bp DNA fragment was amplified from genome DNA of S. aureus TSTw by PCR. This fragment was cloned into pGEM-7Zf(+) and the recombinant plasmid was transformed into E. coli DH5 alpha. The sequencing result of the recombinant plasmid demonstrated that it contains seb gene with 717 bp (without signal encoding region of 81 bp) which has the same nucleotide sequence as described in literature. The seb gene was cloned into expression vector 7ZTS and was transformed into E. coli JM109 (DE3). The expression level of SEB was as high as 33.3% of the cell total proteins.
Cloning, Molecular ; Enterotoxins ; biosynthesis ; genetics ; Escherichia coli ; genetics ; Genetic Engineering ; Recombinant Proteins ; biosynthesis

The rare codons of a fragment in staphylococcal enterotoxin A gene were turned into the most high usage frequency codons in E. coli by overlap PCR technique. Genes of sea and seam were cloned into 7ZTS expression vector and transformed into JM109(DE3), respectively. The result shows that expression level of sea gene was very low, but the expression level of seam was as high as 15% of total cell proteins. The expression product shows activity of antitumor in vivo.
Animals ; Base Sequence ; Codon ; genetics ; Electrophoresis, Polyacrylamide Gel ; Enterotoxins ; genetics ; metabolism ; pharmacology ; Escherichia coli ; genetics ; Gene Expression Regulation, Bacterial ; Male ; Mice ; Molecular Sequence Data ; Neoplasms, Experimental ; drug therapy ; pathology ; Point Mutation ; Recombinant Proteins ; genetics ; metabolism ; pharmacology

OBJECTIVETo identify whether SH3GL1 gene serves as a disease associated gene of adolescent idiopathic scoliosis (AIS).

METHODSPositioning candidate cloning: "case-sibling or case-family control design" research scheme based on family constellation was designed. Fifty-six AIS patients (15 male and 41 female, mean age 15 years old, ranged from 8 to 22 years old, Cobb angle from 25 degrees to 110 degrees , average Cobb angle of 67.5 degrees ) from November 2007 to December 2008 were recruited. In all patients, blood preparation was collected, and genome DNA was extracted. According to nucleotide sequence of gene SH3GL1, primer pair for PCR amplification, cloning, and sequencing with 10 exons as emphasis was designed. Sequence comparative analysis for exon sequencing result between sib pairs or family pairs, and that between sib pair or family pairs and NCBI (National Center for Biotechnology Information) were conducted through Vector NTI Advance 10.3 software to judge whether basic group mutation occurred or not. Amino acid sequence comparative analysis for prediction was made.

RESULTSTen exons of the candidate gene SH3GL1 were successfully amplified and cloned in genome DNA of an AIS sib pair and family pairs, and the sequencing obtained positive results. Twelve basic group mutations were found in 10 exons of the candidate gene SH3GL1 of patients with AIS. These mutations were located in the second exon (3 mutations), the fourth exon (1 mutations), the fifth exon (4 mutations), the sixth exon (1 mutations), the eighth exon (1 mutations), and the tenth exon (2 mutations, noncoding region). If basic group in 515 of mRNA was mutated to T, termination codon(TAG) came into being and open reading frame was altered. The sequence of protein showed brachytmema protein was encoded, which could cause changes of primary structure.

CONCLUSIONSH3GL1 is possibly one of the disease associated genes of AIS.

Adolescent ; Base Sequence ; Child ; Exons ; genetics ; Female ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; Male ; Scoliosis ; genetics ; Sequence Alignment ; Young Adult

OBJECTIVETo compare the tubulogenesis capability of malignant glioma-derived microvessel endothelial cells (GDMEC) from human brain with that of ECV304 cells in a three dimentional model and to explore the significance of GDMEC in the study on angiogenesis.

METHODSThe GDMEC were isolated from malignant gliomas of human brain and purified by selective binding to the monoclonal antibody against CD105 bound to the magnetic MACS MicroBeads. GDMEC and endothelial-like cell line ECV304 were compared with their capabilities of formatting tubule-like structure (TLS) in the three dimentional collagen matrix, with or without inducement by various concentration of vascular endothelial growth factor (VEGF).

RESULTSThe obtained GDMEC had a high purification (98%) and could be successfully cultured in vitro. GDMECs formed more TLS than ECV304 cells of the same number and at the same time points. VEGF could induce rapid formation of TLS in a dose-dependent manner, however, ECV304 cells were less response to VEGF stimulation.

CONCLUSIONSGDMEC could maintain their endothelial characteristics and potential capability of angiogenesis. They were more response to VEGF than ECV304, therefore, more suitable for in vitro studies on tumor angiogenesis.

Brain Neoplasms ; blood supply ; Cells, Cultured ; Dose-Response Relationship, Drug ; Endothelial Cells ; drug effects ; Endothelium, Vascular ; cytology ; Glioma ; blood supply ; Humans ; Immunomagnetic Separation ; Microcirculation ; pathology ; Neovascularization, Pathologic ; Vascular Endothelial Growth Factors ; administration & dosage ; pharmacology

OBJECTIVETo study the morphologic characteristics, immunophenotype and differential diagnosis of a case of microcystic/reticular schwannoma occurring in cervical spine.

METHODSThe pathologic features and immunophenotypic profile of a case of microcystic/reticular schwannoma were studied. Immunohistochemistry was performed using EnVision two-step method.

RESULTSThe patient was a 35-year-old male and presented with a bump over the fifth cervical spine on radiologic check up. Grossly, the bump was gray-white in color, soft, well-circumscribed but non-encapsulated. The tumor measured 3.5 cm × 3.0 cm × 1.8 cm in size. Histologically, it was composed of two distinctive components. One component resembled the conventional schwannoma but showed focally nuclear pleomorphism, reminiscent of changes in degenerating schwannoma. The other component consisted of epithelial-like cells arranged in a reticular or lace-like pattern, amongst a myxoid matrix. Immunohistochemical study showed that the tumor cells were strongly positive for vimentin, S-100 protein, glial fibrillary acidic protein and neuron-specific enolase, focally positive for CD68, CD10 and Ki-67, and negative for pan-cytokeratin, epithelial membrane antigen, neurofilament, carcinoembryonic antigen, smooth muscle actin, estrogen receptor, progesterone receptor and p53.

CONCLUSIONSMicrocystic/reticular schwannoma is a novel variant of schwannoma, arising mainly in internal viscera but seldom in bone. Awareness of this entity is helpful in distinction from chordoma, other mucoid tumors or sarcomas.

Adult ; Cervical Vertebrae ; Chondrosarcoma ; metabolism ; pathology ; Chordoma ; metabolism ; pathology ; Diagnosis, Differential ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Male ; Neurilemmoma ; metabolism ; pathology ; surgery ; Phosphopyruvate Hydratase ; metabolism ; S100 Proteins ; metabolism ; Sarcoma ; metabolism ; pathology ; Spinal Neoplasms ; metabolism ; pathology ; surgery ; Vimentin ; metabolism

OBJECTIVETo investigate the correlation of the testis volume and reproductive hormone level with the results of testicular sperm aspiration (TESA) in non-obstructive azoospermia (NOA) patients, and to explore the cut-off value of the testis volume and reproductive hormone level in predicting the results of TESA so as to provide reliable information for the diagnosis and treatment of NOA.

METHODSWe enlisted 121 NOA patients in this study, divided them into a sperm group and a non-sperm group based on the results of TESA, and measured their testis volumes and reproductive hormone levels.

RESULTSThe left testis volume, the right testis volume, and the levels of prolactin (PRL), follicle-stimulating hormone (FSH), luteinising hormone (LH), estradiol (E2) and total testosterone (T) in the non-sperm and sperm groups were (7.07 +/- 1.06) ml vs (11.75 +/- 1.38) ml, (7.37 +/- 1.37) ml vs (11.70 +/- 1.98) ml, (12.43 +/- 11.69) ng/ml vs (9.60 +/- 4.55) ng/ml, (15.77 +/- 10.84) mIU/ml vs (8.01 +/- 7.43) mIU/ml, (6.12 +/- 2.92) mIU/ml vs (8.11 +/- 20.11) mIU/ml, (119.36 +/- 43.52) pmol/L vs (141.12 +/- 48.33) pmol/L, and (11.43 +/- 4.05) nmol/L vs (12.46 +/- 4.60) nmol/L, respectively. The mean levels of serum FSH and PRL were significantly higher in the non-sperm than in the sperm group. Although the mean testis volume of the former was less than that of the latter, there were no significant differences between the two groups, and nor were any significant differences in age and the levels of E2 and T. The cut-off value of the testis volume was 9 ml, with sensitivity of 93.8%/89.6% (left/right) and specificity of 100%/94.3% (left/right). The area under curve (AUC) of the left testis volume was 0.984 and that of the right was 0.961, indicating a high diagnostic accuracy. The cut-off value of the serum FSH level was 8.18 mIU/ml, with a sensitivity of 71.2% and a specificity of 75.0%. The AUC of the FSH level was 0.743, suggestive of a moderate diagnostic accuracy.

CONCLUSIONThe testis volume and FSH level are important for predicting the TESA results of NOA patients, and the former has even a higher diagnostic accuracy than the latter.

Adult ; Azoospermia ; pathology ; physiopathology ; Follicle Stimulating Hormone ; analysis ; Humans ; Luteinizing Hormone ; analysis ; Male ; Organ Size ; Sperm Retrieval ; Testis ; anatomy & histology ; Young Adult

OBJECTIVETo explore the characteristics on molecular epidemiology of Mycobacterium tuberculosis isolates from hospitals in Tianjin area.

METHODSOne hundred M. tuberculosis isolated strains were collected in succession from August 16th-December 25th, 2005 in Tianjin Haihe Hospital and genotyped by spoligotyping and multiple locus variable number-tandem repeat(VNTR). Data was analyzed by cluster software. Based on the concept of Beijing lineage, it was determinate two sub-groups: atypical Beijing strains and W strain/typical family strains by multiplex and real-time PCR. The associations of subgroups with drug resistance and age were assessed by the chi2 test.

RESULTS96 M. tuberculosis strains were genotyped in which 91.7% (88/96) strains belonged to Beijing genotype (including 3 Beijing-like strains) by spoligtyping. VNTR typing could differentiate 60 genotypes among the 88 Beijing genotype strains. 93.2% of the Beijing lineage M. tuberculosis strains of this study belonged to W strain/typical Beijing family strains (82/88). No statistically significant differences were observed in the proportions of the two sub-groups in patients of different age, or drug resistance (P > 0.05).

CONCLUSIONThe M. tuberculosis Beijing genotype strains were dominated on tuberculosis hospital patients of Tianjin area. The discriminatory power of VNTR typing was higher than that of spoligtyping. The two sub-groups of Beijing lineage had been prevalent in Tianjin, however W strain/typical Beijing family strains were of preponderance.

Bacterial Typing Techniques ; China ; epidemiology ; Cluster Analysis ; DNA, Bacterial ; genetics ; Drug Resistance, Bacterial ; Genotype ; Humans ; Minisatellite Repeats ; Molecular Epidemiology ; Mycobacterium tuberculosis ; classification ; genetics ; isolation & purification ; Sequence Analysis, DNA ; Tuberculosis ; epidemiology