The glutamate dehydrogenase (EC.1.4.1.4) gene which amplified from the genome of Brevibacterium flavum GDK-9 by polymerase chain reaction was linked with pUCm-T for sequence alignment. Analysis of gdh sequences revealed that the whole sequence is 1927 bp, only one ORF existed, which used ATG as the initiation codon and coded a peptide of 448 amino acids with a calculated molecular weight of 48 kD. The comparability between the cloned gdh sequence to the reported sequence is high to 99.55%. Only the 1190th base mutation (C→A) lead to the change of amino acid sequence (Thr→Asn), the others are not. The recombinant plasmid pXG was then transformed into E. coli XL-Blue and Brevibacterium flavum GDK-9 which was induced by IPTG. SDS-PAGE analysis revealed that there was a clear induced protein band with molecular mass of 48.7 kD on expected position. Standard glutamate fermentations indicated that although the level of GDH increases the intracellular glutamate pool, the level of GDH has no influence on glutamate secretion.

OBJECTIVETo study modified ilioinguinal approach through the retrospective analysis on the surgical treatment of 63 patients with pelvic and acetabular fractures through anterior approach.

METHODSFrom January 2006 to January 2013, 63 patients with pelvic and acetabular fractures were treated with the ilioinguinal anterior approach, including 45 males and 18 females, ranging in age from 12 to 68 years old, with an average of (37.71 +/- 13.41) years old. All the patients were divided into two groups: standard ilioinguinal anterior approach group (group A) and modified ilioinguinal anterior approach group(group B). In group A, there were 26 males and 11 females, with an average age of (38.49 +/- 13.64) years old. In group B, there were 19 males and 7 females, with an average age of (36.62 +/- 13.29) years old. Intraoperative and postoperative indicators in group A and B were observed and compared, including operation incision exposure time (from skin incision to complete the ilioinguinal in front of three "windows"), the blood loss, incision close time and treatment effect of Majeed function score.

RESULTSCompared to group A, the incision exposure time of patients in group B was shorter, the blood loss (bleeding during exposure process) was less, and the close incision time was shorter, but the treatment effect of Majeed function score had no significant differences between two groups. All the patients were followed up, and the during ranged from 3 to 36 months, with an average of (18.6 +/- 9.2) months. According to Matta standard assessment reduction of pelvic and acetabular fracture, there were 28 patients got an excellent result, 8 good, and 1 fair in the group A; and 20 patients got an excellent result, 5 good, and 1 fair in the group B. According to Majeed function score for hip function, 20 patients got a satisfactory result, 12 good,4 fair and 1 poor in group A, and the mean score was 82.51 +/- 9.72; and 13 patients got an satisfactory result, 10 good, 3 fair and 0 poor in group B, and the mean score was 80.54 +/- 10.79.

CONCLUSIONThe modified approach has several advantages as follows: providing a good surgical exposure; preventing from the injury of femoral nerve, femoral artery and vein under the inguinal ligament; not needing to open the inguinal canal, which can avoid the occurrence of inguinal hernia, reduce operation prodedures and shorten operation time.

Acetabulum ; injuries ; surgery ; Adolescent ; Adult ; Aged ; Case-Control Studies ; Child ; Female ; Fracture Fixation, Internal ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome ; Young Adult

OBJECTIVETo explore the effects of Bisphenol A in adult rats and its possible mechanisms.

METHODSBPA (in corn oil) was administered orally to 9-week-old male Sprague-Dawley rats for 14 days (0, 1 and 5 g/kg bw), and incubated primary Sertoli cells from pubertal SD rats with 0, 10(-7), 10(-6), 10(-5), 10(-4) mol/L BPA.

RESULTSAfter oral administration, a significant decrease in right testis weight was observed in 5 g/kg dose group, but not in the 1 g/kg bw dose group. Germ cells were detached from basement membrane of seminiferous tubules and Sertoli cells in BPA-treated groups. Administration of BPA at 1 g/kg bw and 5 g/kg bw produced both nucleus pycnosis and vacuolized nucleus in germ cells and Sertoli cells. A marked loss in vimentin staining in Sertoli cells from testis of BPA-treated rats was detected. No change in levels of serum estradiol and testosterone was observed after two-week exposure to BPA. In Sertoli cell primary culture, BPA destroyed the cytoskeleton and cell-cell junctions, and elongated Sertoli cells.

CONCLUSIONThese results suggest that BPA may injure reproductive function of male rats by destroying the cytoskeleton and changing the form of Sertoli cells.

Animals ; Benzhydryl Compounds ; Cells, Cultured ; Cytoskeleton ; drug effects ; Male ; Organ Size ; drug effects ; Phenols ; toxicity ; Rats ; Rats, Sprague-Dawley ; Sertoli Cells ; cytology ; drug effects ; Testis ; anatomy & histology ; cytology ; drug effects ; Vimentin ; metabolism

There was a slow-relaxing tail of skeletal muscles in vitro upon the inhibition of Ca(2+)-pump by cyclopiazonic acid (CPA). Herein, a new linearly-combined bi-exponential model to resolve this slow-relaxing tail from the fast-relaxing phase was investigated for kinetic analysis of the isometric relaxation process of Bufo gastrocnemius in vitro, in comparison to the single exponential model and the classical bi-exponential model. During repetitive stimulations at a 2-s interval by square pulses of a 2-ms duration at 12 V direct currency (DC), the isometric tension of Bufo gastrocnemius was recorded at 100 Hz. The relaxation curve with tensions falling from 90% of the peak to the 15th datum before next stimulation was analyzed by three exponential models using a program in MATLAB 6.5. Both the goodness of fit and the distribution of the residuals for the best fitting supported the comparable validity of this new bi-exponential model for kinetic analysis of the relaxation process of the control muscles. After CPA treatment, however, this new bi-exponential model showed an obvious statistical superiority for kinetic analysis of the muscle relaxation process, and it gave the estimated rest tension consistent to that by experimentation, whereas both the classical bi-exponential model and the single exponential model gave biased rest tensions. Moreover, after the treatment of muscles by CPA, both the single exponential model and the classical bi-exponential model yielded lowered relaxation rates, nevertheless, this new bi-exponential model had relaxation rates of negligible changes except much higher rest tensions. These results suggest that this novel linearly-combined bi-exponential model is desirable for kinetic analysis of the relaxation process of muscles with altered Ca(2+)-pumping activity.
Animals ; Bufonidae ; physiology ; Electric Stimulation ; Electrophysiology ; In Vitro Techniques ; Indoles ; pharmacology ; Kinetics ; Models, Animal ; Models, Biological ; Muscle Relaxation ; drug effects ; physiology ; Muscle Tonus ; drug effects

OBJECTIVETo assess the value of transrectal ultrasonography (TRUS) in the diagnosis of midline prostatic cysts.

METHODSWe retrospectively analyzed the TRUS manifestations of 87 cases of midline prostatic cysts.

RESULTSOf the total number, 33 cases were diagnosed as Müllerian duct cysts, 21 cases ejaculatory duct cysts and the other 33 cases undifferentiated midline prostatic cysts; 19 cases had dilated seminal vesicles, 19 seminal vesicle agenesis, 9 seminal vesiculitis and 5 dilation of the ejaculatory duct.

CONCLUSIONTRUS, convenient, sensitive, safe and non-invasive, is a desirable method for the diagnosis of midline prostatic cysts.

Adolescent ; Adult ; Cysts ; diagnosis ; diagnostic imaging ; Humans ; Male ; Middle Aged ; Prostatic Diseases ; diagnosis ; diagnostic imaging ; Rectum ; Reproducibility of Results ; Sensitivity and Specificity ; Ultrasonography ; methods

OBJECTIVETo determine whether circulating level of catestatin (CST) could provide prognostic information independently of conventional risk markers for the development of in-hospital heart failure in patients with ST-segment elevation myocardial infarction (STEMI).

METHODSThe data of 120 STEMI patients (mean age: 61 years, 73% male) were collected from the Second Hospital of Shanxi Medical University and Taiyuan Central Hospital between November 2010 and September 2011.The patients were categorized into 4 groups according to CST (ng/L) quartile: ≤ 74.72, 74.73-79.67, 79.68 - 84.21 and ≥ 84.22 ng/L. Clinical features, therapeutic approaches were compared among groups. The patients were also grouped according to Killip class: Killip level I (n = 68), Killip level II (n = 23), Killip level III (n = 18), Killip level IV (n = 11). CST, NE and NT-proBNP were compared among groups. The Spearma rank correlation and multivariate logistic regression analysis were applied to determine the association between risk factors and in-hospital heart failure. Receiver-operator characteristic (ROC) curve was performed to evaluate the power of CST and NT-proBNP on predicting in-hospital heart failure.

RESULTSGender, hospital days, past history of smoking, hypertension, myocardial infarction, CK-MB peak level, TnI peak level, heart rate, blood pressure, blood glucose, blood lipid levels on admission and early reperfusion therapy were similar among groups. Patients with higher CST values were more likely to be older, to have lower body mass index, to have higher white blood cell count, CysC, hs-CRP, NE, NT-proBNP, past history of angina, diabetes mellitus, being diuretic users, and to have a lower ejection fraction (all P < 0.05). Higher CST levels were also associated with increased risk of heart failure (P < 0.05). In proportion with the deterioration of the cardiac function, CST, NE, NT-proBNP concentration gradually increased (all P < 0.05). Spearman rank correlation analysis showed that the CST was negatively correlated with LVEF (r(s) = -0.923, P < 0.001) and positively correlated with NT-proBNP (r(s) = 0.884, P < 0.001). After multivariate adjustment, CST remained to be an independent risk factor for the development of in-hospital heart failure (OR = 1.125, 95%CI: 1.056 - 1.198;P < 0.001). The area under the ROC curve of CST and NT-proBNP was 0.777 and 0.874. Using CST = 77.29 ng/L as a cut-off value, the sensitivity was 92.8% and specificity was 70.6% for predicting the development of in-hospital heart failure.

CONCLUSIONThe plasma CST level is an independent predictor for the development of in-hospital heart failure in patients with STEMI.

Aged ; Catechols ; antagonists & inhibitors ; pharmacology ; Chromogranin A ; blood ; Female ; Heart Failure ; diagnosis ; etiology ; Humans ; Inpatients ; Male ; Middle Aged ; Myocardial Infarction ; blood ; complications ; diagnosis ; Peptide Fragments ; blood ; Prognosis ; Prospective Studies ; Risk Factors

OBJECTIVETo analyze the promoter methylation levels of p15, CDH1, DAPK and HICI genes of patients with myelodysplastic syndrome (MDS) and explore the relationship between the level of methylation and clinical features.

METHODSDNA methylation levels of p15, CDH1, DAPK and HICI in peripheral blood (PB) or bone marrow (BM) samples from 52 MDS patients were detected by real-time quantitative PCR. The correlation of the methylation level with clinical features and hematological findings was analyzed. 38 de novo AML patients and 46 normal individuals served as controls.

RESULTSThe methylation levels of p15, CDH1, DAPK and HICI were 16.23 ± 21.69, 6.59 ± 9.39, 0.14 ± 0.11 and 7.81 ± 9.70 in BM, and 14.96 ± 20.16, 6.00 ± 9.26, 0.12 ± 0.14 and 6.74 ± 9.72 in PB, respectively from 18 MDS patients, and the difference between BM and PB was not statistically significant (P > 0.05). The methylation levels of p15 (14.70 ± 18.17) and CDH1 (6.61 ± 8.79) genes in high risk (RAEBI/II) MDS were significantly higher than in low risk (RCMD/RARS/5q-, p15: 1.99 ± 1.59, CDH1: 1.23 ± 1.14 and RCMD, p15: 3.02 ± 3.42, CDH1:1.53 ± 2.06) MDS or control (p15: 1.69 ± 1.82, CDH1: 1.01 ± 1.12) (P < 0.05). The methylation levels of DAPK gene had no difference among subtypes of MDS, and that of HIC1 gene only differed between RAEB I/II (9.16 ± 11.95) and control (2.49 ± 2.26) (P = 0.042). The difference of methylation levels of p15, CDH1, DAPK and HICI in BM was statistically significant among subtypes of MDS (P = 0.001, 0.003, 0.039, 0.023, respectively). And so did of p15 and DAPK in PB (P = 0.013, 0.006, respectively). The methylation level of p15 and CDH1 was significantly correlated with IPSS classification and blasts percentage in BM.

CONCLUSIONSp15 and CDH1 genes are special hypermethylation genes in MDS. Methylation level of HIC1 gene showed an upward tendency from low risk to high risk MDS.

Adult ; Aged ; Aged, 80 and over ; Apoptosis Regulatory Proteins ; genetics ; metabolism ; Cadherins ; genetics ; metabolism ; Calcium-Calmodulin-Dependent Protein Kinases ; genetics ; metabolism ; Case-Control Studies ; Cyclin-Dependent Kinase Inhibitor p15 ; genetics ; metabolism ; DNA Methylation ; Death-Associated Protein Kinases ; Female ; Humans ; Kruppel-Like Transcription Factors ; genetics ; metabolism ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; metabolism ; Promoter Regions, Genetic ; Real-Time Polymerase Chain Reaction ; Young Adult

OBJECTIVETo assess the prognostic value of biological features and therapy-related factors in multiple myeloma (MM).

METHODS123 patients with newly diagnosed MM between January 1998 and May 2005 were enrolled in this retrospective study. Biological features at presentation and therapy-related factors were analysed. The overall survival (OS) and time to progression (TTP) were estimated by Kaplan-Meier analysis and the distribution of OS and TTP were compared using log-rank test. Cox regression was used to identify the independent prognostic factors.

RESULTS(1) The univariate analysis indicated that more immature plasma cells in bone marrow biopsy, C-reactive protein >8. Omg/L, CD117 expression, serum beta2-microglobulin (beta2-MG) (3.5 approximately 5.5 mg/L), abnormal cytogenetics aberration of chromosome 13 (Delta13), hypodiploid, poor response to chemotherapy, interferon(IFN) therapy less than 6 months were associated with shorter OS(P <0.05). Lytic bone lesions at presentation, more immature plasma cells in bone marrow biopsy, serum beta2-MG (3.5 approximately 5.5 mg/L), poor response to chemotherapy, and IFN therapy less than 6 months as well as abnormal cytogenetics, hypodiploid and Delta13 were associated with shorter TTP (P <0.05). (2) Multivariable COX analysis indicated IFN therapy more than 6 months was a protective factor for OS and TTP, and more immature plasma cells in bone marrow biopsy was an independent poor prognostic factor for TTP.

CONCLUSIONThe morphology of myeloma cells is useful for assessing the prognosis. And IFN therapy more than 6 months could lengthen OS and TTP.

Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; diagnosis ; pathology ; therapy ; Prognosis ; Retrospective Studies ; Risk Factors ; Survival Analysis

The discovery of induced pluripotent stem cells(iPSCs) is a promising advancement in the field of regenerative medicine. Previous studies have indicated that the teratoma-forming propensity of iPSCs is variable; however, the relationship between tumorigenic potential and genomic instability in human iPSCs (HiPSCs) remains to be fully elucidated. Here, we evaluated the malignant potential of HiPSCs by using both colony formation assays and tumorigenicity tests. We demonstrated that HiPSCs formed tumorigenic colonies when grown in cancer cell culture medium and produced malignancies in immunodeficient mice. Furthermore, we analyzed genomic instability in HiPSCs using whole-genome copy number variation analysis and determined that the extent of genomic instability was related with both the cells' propensity to form colonies and their potential for tumorigenesis. These findings indicate a risk for potential malignancy of HiPSCs derived from genomic instability and suggest that quality control tests, including comprehensive tumorigenicity assays and genomic integrity validation, should be rigorously executed before the clinical application of HiPSCs. In addition, HiPSCs should be generated through the use of combined factors or other approaches that decrease the likelihood of genomic instability.
Animals ; Carcinogenesis ; Cells, Cultured ; DNA Copy Number Variations ; Genomic Instability ; Humans ; Induced Pluripotent Stem Cells ; cytology ; metabolism ; transplantation ; Mice ; Mice, SCID ; NIH 3T3 Cells ; Octamer Transcription Factor-3 ; metabolism ; Teratocarcinoma ; etiology ; Teratoma ; etiology ; Tumor Stem Cell Assay

With the increasing cost of drug development and clinical trials, it is of great value to make full use of all kinds of data to improve the efficiency of drug development and to provide valid information for medication guidelines. Model-based meta-analysis (MBMA) combines mathematical models with meta-analysis to integrate information from multiple sources (preclinical and clinical data, etc.) and multiple dimensions (targets/mechanisms, pharmacokinetics/pharmacodynamics, diseases/indications, populations, regimens, biomarkers/efficacy/safety, etc.), which not only provides decision-making for all key points of drug development, but also provides effective information for rational drug use and cost-effectiveness analysis. The classical meta-analysis requires high homogeneity of the data, while MBMA can combine and analyze the heterogeneous data of different doses, different time courses, and different populations through modeling, so as to quantify the dose-effect relationship, time-effect relationship, and the relevant impact factors, and thus the efficacy or safety features at the level of dose, time and covariable that have not been involved in previous studies. Although the modeling and simulation methods of MBMA are similar to population pharmacokinetics/pharmacodynamics (Pop PK/PD), compared with Pop PK/PD, the advantage of MBMA is that it can make full use of literature data, which not only improves the strength of evidence, but also can answer the questions that have not been proved or can not be answered by a single study. At present, MBMA has become one of the important methods in the strategy of model-informed drug development (MIDD). This paper will focus on the application value, data analysis plan, data acquisition and processing, data analysis and reporting of MBMA, in order to provide reference for the application of MBMA in drug development and clinical practice.