Role of OCT2 and MATE1 in renal disposition and toxicity of nitidine chloride.

BACKGROUND AND PURPOSE: Nitidine chloride (NC), a benzophenanthridine alkaloid, has various biological properties including anticancer and analgesic activities. The aim of the present study was to evaluate the role of organic cation transporter 2 (OCT2) and multidrug and toxin extrusion 1 (MATE1) in the renal disposition and nephrotoxicity of NC. EXPERIMENTAL APPROACH: MDCK cells stably expressing human OCT2 and/or hMATE1 were used to investigate the OCT2- and MATE1-mediated transport of NC. In addition, the accumulation of NC and its potential toxicity were studied in rat primary-cultured proximal tubular (rPCPT) cells and in rats in vivo. KEY RESULTS: NC was found to be a high-affinity substrate of both OCT2 and MATE1 with high cytotoxicity in MDCK-hOCT2/hMATE1 and MDCK-hOCT2 compared to mock cells. The OCT2 inhibitors, cimetidine and (+)-tetrahydropalmatine ((+)-THP), significantly reduced NC accumulation and cytotoxicity in MDCK-hOCT2, MDCK-hOCT2/hMATE1 and rPCPT cells. Severe kidney damage with high levels of blood urea nitrogen and lactate dehydrogenase (LDH), reduced levels of alkaline phosphatase (ALP) and pathological changes were found in rats after 20 days of successive i.v. doses of NC (5 mg·kg(-1) ·day(-1) ). Concomitantly, the concentration of NC in the kidney reached similar high levels at 2 h after the last dose of the 20 day treatment as those observed at 0.5 h after a single i.v. dose of 5 mg·kg(-1) . CONCLUSIONS AND IMPLICATIONS: Our data indicate that NC-induced nephrotoxicity might be mainly attributed to OCT2-mediated extensive renal uptake and weak tubular secretion by MATE1.

[Value of the albumin to globulin ratio in predicting severity and prognosis in myasthenia gravis patients].

OBJECTIVE: To assess the predictive value of the albumin to globulin ratio (AGR) in evaluation of disease severity and prognosis in myasthenia gravis patients. METHODS: A total of 135 myasthenia gravis (MG) patients were enrolled between February 2009 and March 2015. The AGR was detected on the first day of hospitalization and ranked from lowest to highest, and the patients were divided into three equal tertiles according to the AGR values, which were T1 (AGR <1.34), T2 (1.34≤AGR≤1.53) and T3 (AGR>1.53). The Kaplan-Meier curve was used to evaluate the prognostic value of AGR. Cox model analysis was used to evaluate the relevant factors. Multivariate Logistic regression analysis was used to find the predictors of myasthenia crisis during hospitalization. RESULTS: The median length of hospital stay for each tertile was: for the T1 21 days (15-35.5), T2 18 days (14-27.5), and T3 16 days (12-22.5) (P<0.01), and Kaplan-Meier curves showed significant difference among the three groups. In the univariate model, serum albumin, creatinine, AGR and MGFA clinical classification were related to prognosis of myasthenia gravis. At the multivariate Cox regression analysis, the AGR (P<0.001) and MGFA clinical classification (P<0.001) were independent predictive factors of disease severity and prognosis in myasthenia gravis patients. Respectively, the hazard ratio (HR) were 4.655 (95% CI: 2.355-9.202) and 0.596 (95% CI: 0.492-0.723). Multivariate Logistic regression analysis showed the AGR (P<0.001) and MGFA clinical classification were related to myasthenia crisis. CONCLUSION: The AGR may represent a simple, potentially useful predictive biomarker for evaluating the disease severity and prognosis of patients with myasthenia gravis.

[Dose optimization of lipid infusionin treatment of patients with acute dexmedetomidinepoisoning].

OBJECTIVE: To optimize the dose of lipid infusion in treatment of patients with acute dexmedetomidinepoisoning, in order to further guide the rational use of medication in clinical practice. METHODS: A total of 80 patients with acute dexmedetomidinepoisoning were admitted in this study from January 2012 to October 2014 at our hospital and divided into three groups based on the intensity of poisoning, including: slight poisoning (28 cases), moderate poisoning (32 cases) and severe poisoning (20 cases). Patients in each group were given 10% lipid infusion or 20% lipid infusion for treatment.Stable blood dexmedetomidineconcentrations of patients in pre-treatment and at different time points after treatment (pre-treatment and 0.5, 1, 2, 5, 10, 20 h after treatment) and the length of hospital stay, awake time in each group were investigated and compared.Ramsay sedation scores were recorded and compared in different time points (0.5 h before treatment and 2, 5, 20 h after treatment) in each group for different treatments.Side effects and complications were recorded, and follow-up was conducted during 1-3 d post discharge to record the recovery condition in patients. RESULTS: In each group, patients receiving 20% lipid infusion waked earlier than those receiving 10% lipid infusion.And the hospitalization duration for patients receiving 20% lipid infusion was significantly shorter than those receiving 10% lipid infusion [(4.6±1.6) h vs (6.7±2.0) h, (2.6±0.4) d vs (4.0±0.6) d, P<0.05]. The Ramsay sedation scores were significantly lower for patients receiving 20% lipid infusion than those receiving 10% lipid infusionat 2 h and 5 h after treatment in each group [(3.4±0.3) vs (4.7±0.4), (2.6±0.3) h vs (3.5±0.3) h, P<0.05]. The stable plasma concentrations of dexmedetomidine were gradually reduced after the treatment, and which were lower when compared with the theoretical metabolic concentration.What's more, the plasma concentrationsat 1 h, 2 h and 5 h after treatment were significantly lower for patientsreceiving 20% lipid infusion than those receiving 10% lipid infusion in each group (P>0.05). All patients in our study were cured and discharged without severe side effects and complications, and follow-ups showed that no patients showed evidence of rebound phenomenon. CONCLUSIONS: Different concentrations of lipid infusionare safe and effective in relieving the intensity of dexmedetomidinepoisoning, and promoting the clinical recovery.What's more, the therapeutic efficacy of 20% lipid infusion is greater than 10% lipid infusion.

[Application of rabbit monoclonal antibody GCET2 in diagnosis of diffuse large B cell lymphoma].

Objective: To prepare a rabbit monoclonal antibody GCET2 and to investigate its diagnostic value in the workup of diffuse large B-cell lymphoma (DLBCL). Methods: GCET2 rabbit monoclonal antibody was developed by using RabMAb® technology, and its specificity was confirmed by ELISA, Western blot, immunohistochemistry (IHC) and flow cytometry. A panel of immunomarkers including GCET2, CD10, bcl-6, MUM1, GCET1, FOXP1, Ki-67 and CMYC was evaluated in 81 cases of DLBCLs, 5 cases of follicular lymphomas (FL) and 2 cases of Burkitt's lymphomas. Results: Rabbit monoclonal GCET2 antibody (clone EP316) was developed with specificity for normal germinal center B-cells (GCB) and GCB origined lymphomas. In 81 cases of DLBCL, the positive rate of GCET2 was 43.2%(35/81), which was significantly higher than that of other germinal center markers. Moreover, among 81 DLBCLs, the proportions of high CMYC expression in GCET2 negative and positive groups were 15.2% (7/46) and 2.8% (1/35), respectively. Conclusion: GCET2 is a sensitive immunohistochemical marker for GCB derived lymphomas. Combined with other biomarkers, it may improve the diagnostic sensitivity of GCB-DLBCL.

Absence of mutations in human ubiquitin fusion-degradation protein gene in tetralogy of Fallot.

Congenital defects in human chromosome 22q11 deletion syndromes are associated with the 3rd and 4th pharyngeal pouch during fetal development. In the cardiovascular system, these disorders are usually apparent as conotruncal heart defects and aortic arch anomalies. UFD1L, a gene that is downregulated in dHAND-deficient mice, expressed in the mouse embryo at the branchial arch and mapped to human chromosome 22q11, has recently been strongly suspected to be responsible for the phenotypes expressed in 22q11 deletion syndromes. Its putative causal role in relevant congenital cardiovascular malformations was studied by gene dosage analysis, mutation screening and sequence analyses. Sixty cases of tetralogy of Fallot with no detectable chromosome deletion at 22q11 or 10p13 were examined, including 51 cases of simple tetralogy of Fallot, and 9 cases of tetralogy of Fallot with pulmonary atresia. None of these patients revealed deletion limited to a portion of the UFD1L gene. Although mobility shift was found by heteroduplex analysis in 24 cases at exon 4 and flanking sequences, further sequence analysis demonstrated only two silent nucleotide variations and a single nucleotide polymorphism in intron 4. Our data suggest that, although the UFD1L gene is mapped to 22q11 and is expressed during early murine development at both cardiac and cranial neural crests, it is not responsible for the majority of tetralogy of Fallot cases in humans.