[Function of MEIS1 and miR-425 to the Regulating of Chronic Myeloid Leukemia Cell Proliferation].

OBJECTIVE: To investigate the function and mechanism of transcription factor of MEIS1 and miR-425 to the proliferation of chronic myeloid leukemia cell K562. METHODS: Bioinformatic prediction was used to analyze the binding of MEIS1 in miR-425 promoter region. ChIP-qPCR coupled with dual luciferase assay was used to detect the combination of MEIS1 and the transcription activity of miR-425, and its regulative role in the transcription activity miR-425. CCK-8 was used to detect the effect of MEIS1 and miR-425 on cell proliferation. Flow cytometry with PI staining was used to detected the effect of MEIS1 and miR-425 on K562 cell cycle progression. Western blot was used to examine the effect of miR-452 on the expression level of MEIS1. RESULTS: MEIS1 could bind the promoter of miR-425 and repressed its transcription. After K562 was transfected by shRNA, the K562 cell proliferation and cell cycle progression was significantly inhibitied. Moreover, after K562 cells were transfected by miR-425 mimic, cell proliferation and cell cycle was inhibited. The expression level of MEIS1 could be inhibited by the combination of miR-425 and MEIS1 3'UTR. CONCLUSION: MEIS1 can inhibit the activity of miR-425 in transcriptional level, while the miR-425 can suppress the expression of MEIS1 protein in post-transnational level. Therefore, a regulatory circuit comprising from MEIS1 and miR-425 regulates K562 cell proliferation.

Correlation of the leucocyte count with traditional and non-traditional components of metabolic syndrome.

OBJECTIVES: To investigate correlation of the white blood cell (WBC) and its subtype count with the traditional and non-traditional components of the metabolic syndrome. METHODS: Between January 2012 and December 2013, 18,222 people were enrolled in this study. The height, weight, body mass index (BMI) and blood pressure were measured, and blood samples were tested for all subjects after an overnight fast. The count of WBC and its subtypes, total cholesterol, triglyceride, high density lipoprotein (HDL), low-density lipoprotein, aminotransferases, fibrinogen, uric acid, and fasting blood glucose were all assessed. RESULTS: Metabolic syndrome was found in 2502 of 18,222 healthy Chinese people (16.41%). The prevalence of metabolic syndrome was 22.61% for men significantly (P < 0.05) greater than for women (6.83%). The prevalence of obesity, hypertension, hyperglycemia and hyperlipidemia was significantly (P < 0.001) higher in people with than without metabolic syndrome. With increase of the WBC count, BMI, systolic and diastolic pressure, fasting blood glucose, triglyceride, glutamic-oxaloacetic transaminase, glutamic-pyruvic transaminase, glutamyltranspetidase, blood urea nitrogen fibrinogen and uric acid all went up significantly (P < 0.001) while HDL decreased significantly (P < 0.05). The creatinine remained relatively sTable After adjustment of age, sex, alcoholic drinking and education, the metabolic components of obesity, hypertension, diabetes and hyperlipidemia rose significantly (P < 0.05) positively with increased counts of the total WBC, neutrophil and lymphocyte, and the WBC and its subtypes were an independent risk factor for metabolic syndrome. CONCLUSION: Aminotransferases, fibrinogen and uric acid all significantly increase with increased WBC count in a dose-dependent manner. Increased counts of the total WBC and its subtypes are positively associated with presence of metabolic syndrome.

Management of antibiotic-associated pseudomembranous colitis in Non-hospitalized and hospitalized patients.

Proper management of antibiotic-associated pseudo membranous colitis is not clear. This article is to investigate proper treatment of antibiotic-associated pseudo membranous colitis. Data of 67 patients (aged 18-69 years, with 31 males and 46 females) with antibiotic-associated pseudo membranous colitis were retrospectively analyzed including the demography, antibiotics to induce and for treatment of the pseudo membranous colitis, and other supportive measures. All 67 patients had a positive cytotoxin test, which confirmed the pseudo membranous colitis. Antibiotics which induced the pseudo membranous colitis included clindamycin, ofloxacin, piperacillin, cefatriaxone, penbritin and ceftazidime. Once the correct diagnosis was made, the culprit antibiotics were discontinued immediately, and narrow-spectrum antibiotics like metronidazole and vancomycin were administered in combination with correction of fluid and electrolyte abnormalities, use of vitamins C and B complex to repair the intestinal mucosa, and avoidance of antispasmodic and antidiarrheal agents. After appropriate treatment for 2-20 days, all patients recovered with no sequela. Sixty-two patients were clinically cured while five (7.5%) had diarrhea recurrence within two months of the end of therapy. Retreatment with tapering and extended period of metronidazole and/or vancomycin led to complete recovery of the patients. Multiple antibiotic agents are associated with pseudo membranous colitis, and correction of fluid and electrolyte abnormalities and use of vitamins to repair the intestinal mucosa should be performed to speed up the cure process.

MicroRNA-10b is upregulated and has an invasive role in colorectal cancer through enhanced Rhoc expression.

We found that the difference in miR­10b expression between the tumor tissue and adjacent non­tumor tissue was significant. Outer periphery and portal vein serum miR­10b concentrations were significantly higher than those of the control. However, the outer periphery vein miR­10b concentrations were not significant when compared with the portal vein serum concentration in colorectal cancer. The expression levels of miR­10b were associated with higher­grade colorectal cancer. MiR­10b levels were markedly elevated in lymph node metastasis-positive tumor tissue compared with those in lymph node metastasis-free tumor tissue, and were correlated with a downregulation in Hoxd10 expression. Rhoc protein expression in tumor tissue was significantly amplified when compared to that of the control tissue group. An inverse correlation between Hoxd10 and Rhoc in immunohistochemistry and western blot analysis was observed (P<0.05). MiR-10b expression was also inversely correlated with Hoxd10 protein expression (P<0.05). Thus, miR­10b is potentially involved in the invasion of colorectal cancer.

Combined detection tumor markers for diagnosis and prognosis of gallbladder cancer.

AIM: To clarify the value of combined use of markers for the diagnosis of gallbladder cancer and prediction of its prognosis. METHODS: Serum cancer antigens (CA)199, CA242, carcinoembryonic antigen (CEA), and CA125 levels were measured in 78 patients with gallbladder cancer (GBC), 78 patients with benign gallbladder diseases, and 78 healthy controls using electrochemiluminescence. CA199, CA242, CEA, and CA125 levels and positive rates were analyzed and evaluated pre- and post-operatively. Receiver operator characteristic curves were used to determine diagnostic sensitivity and specificity of GBC. Survival time analysis, including survival curves, and multivariate survival analysis of a Cox proportional hazards model was performed to evaluate independent prognostic factors. RESULTS: Serum CA242, CA125, and CA199 levels in the GBC group were significantly higher when compared with those in the benign gallbladder disease and healthy control groups (P < 0.01). With a single tumor marker for GBC diagnosis, the sensitivity of CA199 was the highest (71.7%), with the highest specificity being in CA242 (98.7%). Diagnostic accuracy was highest with a combination of CA199, CA242, and CA125 (69.2%). CA242 could be regarded as a tumor marker of GBC infiltration in the early stage. The sensitivity of CA199 and CA242 increased with progression of GBC and advanced lymph node metastasis (P < 0.05). The 78 GBC patients were followed up for 6-12 mo (mean: 8 mo), during which time serum CA199, CA125, and CA242 levels in the recurrence group were significantly higher than in patients without recurrence (P < 0.01). The post-operative serum CA199, CA125, and CA242 levels in the non-recurrence group were significantly lower than those in the GBC group (P < 0.01). Multivariate survival analysis using a Cox proportional hazards model showed that cancer of the gallbladder neck and CA199 expression level were independent prognostic factors. CONCLUSION: CA242 is a marker of GBC infiltration in the early stage. CA199 and cancer of the gallbladder neck are therapeutic and prognostic markers.

Impact of direct cell co-cultures on human adipose-derived stromal cells and nucleus pulposus cells.

Biologic and cellular treatment strategies aiming for curing intervertebral disc degeneration (IDD) have been proposed recently. Given the convenient availability and expansion potential, adipose-derived stromal cells (ADSCs) might be an ideal cell candidate. However, the interaction between ADSCs and nucleus pulposus (NP) cells still remains ambiguous, especially in direct co-cultures of the two types of cells. Nevertheless, NP markers in ADSCs after co-cultures were unidentified. Here, we addressed the interaction of human ADSCs and NP cells in a direct co-culture system for the first time. As a result, ADSCs could differentiate to the NP cell phenotype with a significant up-regulated expression of multiple genes and proteins in extracellular matrix (ECM) (SOX9, COL2A1, ACAN, and COL6A2), relative NP markers (FOXF1, PAX1, CA12, and HBB) and pertinent growth factors (CDMP-1, TGF-ß1, IGF-1, and CTGF). Moreover, the gene expression of COL2A1, ACAN, and COL6A2 of degenerate NP cells was also up-regulated. Collectively, these results suggest that direct co-cultures of ADSCs and NP cells may exert a reciprocal impact, that is, both stimulating ADSCs differentiation to the NP cell phenotype and inducing NP cells to regain functional phenotype. Accordingly, ADSCs might be a potential candidate in the development of cellular treatment strategies for IDD.

Immune cascades in human intervertebral disc: the pros and cons.

The unique structural hallmark of the intervertebral disc has made its central composition, the nucleus pulposus (NP), excluded from the immunologic tolerance. Consequently, the intervertebral disc is identified as an immune-privileged organ. Traditionally, local detrimental immune activities caused by NP at the lesion sites of the disc are noted as a significant factor contributing to disc degeneration. However, given the beneficial activities of immune cells in other immune-privileged sites on basis of current evidence, the degenerate disc might need the assistance of a subpopulation of immune cells to restore its structure and lessen inflammation. In addition, the beneficial impact of immune cells can be seen in the absorption of the herniated NP, which is an important factor causes the mechanical compression of nerve roots. Consequently, a modulated immune network in degenerate disc is essential for the restoration of this immune-privileged organ. Until now, the understandings of immune response in disc degeneration still rest on the harmful aspect. Further studies are needed to explore its beneficial influence. Accordingly, there are no absolutely the pros and cons in terms of immune reactions caused by NP.

Identification of proteins differentially expressed in adriamycin-resistant (pumc-91/ADM) and parental (pumc-91) human bladder cancer cell lines by proteome analysis.

PURPOSE: Resistance to chemotherapy drugs remains a difficult problem in bladder cancer treatment. Protein expression is an important factor underlying multidrug resistance (MDR) in bladder cancer. The aim of the study was to explore differentially expressed proteins responsible for MDR between an adriamycin-resistant human bladder cancer cell line (pumc-91/ADM) and its parental cell line (pumc-91). METHODS: Two-dimensional gel electrophoresis (2-DE) combining image analysis was used to screen the differentially expressed protein spots between the pumc-91/ADM and pumc-91 cell lines. Then, the protein spots were identified using MALDI-TOF/TOF mass spectrometry. Among the identified proteins, annexin A2 (ANXA2) and nucleophosmin (NPM1) were then further verified using RT-PCR and Western blot analysis. RESULTS: A total of 30 proteins, including 19 up-regulated and 11 down-regulated proteins, were successfully identified in pumc-91/ADM. According to their different functions, these 30 proteins were classified into 12 categories. Annexin A2 (ANXA2) and nucleophosmin (NPM1) were up-regulated in pumc-91/ADM compared with pumc-91. CONCLUSION: The proteins identified may have an important clinical significance in MDR, and ANXA2 and NPM1 may take part in mechanism of MDR in bladder cancer.

[Comparison and evaluation of testing results for two different coagulation analyzers].

OBJECTIVE: To evaluate the performance of a newly installed fully automatic coagulation analyzer and compare the consistency of its testing results with the confirmed clinical automatic coagulation analyzer at our department. METHODS: Precision, linearity, carryover and accuracy of the newly installed coagulation analyzer were evaluated according to the national required standards. Then the testing results were analyzed between the newly installed and confirmed coagulation analyzers according to the EP-5 and EP-9 documents of national committee for clinical laboratory standards (NCCLS). RESULTS: For the newly installed coagulation analyzer, the low, median and high values of relative intra-precision were: 0.93%, 1.32% and 1.27% for prothrombin time (PT); 1.42%, 0.84% and 1.17% for activated partial thromboplastin time (APTT); 1.82%, 3.13% and 3.19% for fibrinogen (FIB); 1.78%, 1.76% and 1.38% for thrombin time (TT) respectively. The linear regression equation of FIB actual and theoretical values was y = 1.012x + 0.0219 (P > 0.05). There was no significant statistical difference between the intercept and 0 (t = 0.2287, P > 0.05) and between linear slope and 1 (t = 0.3221, P > 0.05). The carryover was -2.33%. The testing results of defined acceptable bias of PT and FIB in CLIA'88 for two analyzers were within the acceptable 95% confidence interval of bias. CONCLUSION: The precision, linearity, carryover and accuracy of the newly installed coagulation analyzer meet the requirements of instrument user manual. The performance and the testing results of the same sample from two coagulation analyzers are consistent.

Vibrio vulnificus cytolysin induces apoptosis in HUVEC, SGC-7901 and SMMC-7721 cells via caspase-9/3-dependent pathway.

Vibrio vulnificus cytolysin (VVC) is known to be a pore-forming toxin which shows cytotoxicity for mammalian cells in culture and induces apoptosis in endothelial cells. In order to determine whether VVC induces apoptosis in vascular endothelial cells and tumor cells, the cytotoxicity induced by recombinant VVC (rVVC) and its potential mechanism in HUVEC, SGC-7901 and SMMC-7721 cells were investigated. Our study demonstrated that rVVC induced the release of intracellular K(+) from all the target cells, yet lactate dehydrogenase was not released by rVVC. It indicates that osmotic lysis might not contribute to the cytolysin-induced cytotoxicity. The study also demonstrated that rVVC induced apoptosis in HUVEC, SGC-7901 and SMMC-7721 cells in time- and dosage-dependent manners, which was associated with the activation of caspase-9 and -3, but not caspase-8. During the apoptotic process of the target cells, rVVC labeled with FITC was monitored to attach initially to the surface of the cells and entered the cytoplasma subsequently. These findings suggest that VVC may be not only a pore-forming toxin, but also a transmembrane toxin with powerful ability to induce apoptosis in human vascular endothelial cells and tumor cells.