Population pharmacokinetics of mycophenolic acid in pediatric patients with primary IgA nephropathy / 中国药房

OBJECTIVE To develop a population pharmacokinetic （PPK） model for mycophenolate mofetil active metabolite mycophenolic acid （MPA） in children with primary IgA nephropathy， explore the factors affecting the pharmacokinetic parameters of MPA， and provide a basis for clinical individualized therapy. METHODS Retrospective collection was conducted on 636 concentrations and clinical data from 47 pediatric patients with primary IgA nephropathy. PPK analysis was carried out by using the nonlinear mixed-effects model； the covariates were tested with a stepwise method. Goodness-of-fit plots， Bootstrap and visual predictive check were employed to evaluate the final model. RESULTS The pharmacokinetics of MPA in children with IgA nephropathy in vivo conformed to the first-order absorption and elimination two-compartment model （objective function value of 3 276.31）. Covariate analysis suggested that body weight and albumin （ALB） levels were significant influencing factors on apparent clearance rate and apparent distribution volume. The typical values of PPK parameters of MPA in the final model were as follows： the central room had a distributed volume of 5.79 L， the clearance rate was 4.06 L/h， the volume of peripheral ventricular distribution was 430.93 L， the clearance rate between compartments was 15.40 L/h， the oral absorption rate constant was 1.29 h－1. After verification， most of the predicted corrected observed concentration points were within the 90% confidence interval of the predicted corrected simulated concentration， indicating that the MPA final model had good predictive performance. CONCLUSIONS The PPK model of MPA in children with primary IgA nephropathy is established in this study， identifying body weight and ALB levels are significant factors affecting MPA metabolism.

RITA selectively inhibits proliferation of BAP1-deficient cutaneous melanoma cells in vitro / 南方医科大学学报

OBJECTIVE@#To screen for small molecular compounds with selective inhibitory activity against cutaneous melanoma cells with BAP1 deletion.@*METHODS@#Cutaneous melanoma cells expressing wild-type BAP1 were selected to construct a BAP1 knockout cell model using CRISPR-Cas9 system, and small molecules with selective inhibitory activity against BAP1 knockout cells were screened from a compound library using MTT assay. Rescue experiment was carried out to determine whether the sensitivity of BAP1 knockout cells to the candidate compounds was directly related to BAP1 deletion. The effects of the candidate compounds on cell cycle and apoptosis were detected with flow cytometry, and the protein expressions in the cells were analyzed with Western blotting.@*RESULTS@#The p53 activator RITA from the compound library was shown to selectively inhibit the viability of BAP1 knockout cells. Overexpression of wild-type BAP1 reversed the sensitivity of BAP1 knockout cells to RITA, while overexpression of the mutant BAP1 (C91S) with inactivated ubiquitinase did not produce any rescue effect. Compared with the control cells expressing wild-type BAP1, BAP1 knockout cells were more sensitive to RITA-induced cell cycle arrest and apoptosis (P < 0.0001) and showed an increased expression of p53 protein, which was further increased by RITA treatment (P < 0.0001).@*CONCLUSION@#Loss of BAP1 results in the sensitivity of cutaneous melanoma cells to p53 activator RITA. In melanoma cells, the activity of ubiquitinase in BAP1 is directly related to their sensitivity to RITA. An increased expression of p53 protein induced by BAP1 knockout is probably a key reason for RITA sensitivity of melanoma cells, suggesting the potential of RITA as a targeted therapeutic agent for cutaneous melanoma carrying BAP1-inactivating mutations.

Bortezomib and obatoclax for dual blockade of protein degradation pathways show synergistic anti-tumor effect in human acute T lymphoblastic leukemia cells / 南方医科大学学报

OBJECTIVE@#To explore whether bortezomib and a Bcl-2 inhibitor exhibit synergistic anti-tumor effect in human acute T lymphoblastic leukemia cells.@*METHODS@#MTT assay was used to determine the cytotoxicity of bortezomib in the absence or presence of Bcl-2 inhibitors (obatoclax, AT-101 and ABT-199) in Jurkat cells. The effects of drug treatment on the expression of Bcl-2 family proteins, LC3B, p62, ubiquitin, BiP/Grp78, p-JNK, p-p38 and CHOP proteins were examined by Western blotting. Flow cytometry was used to determine the effects of bortezomib and Bcl-2 inhibitors (obatoclax, AT-101 and ABT-199) on cell apoptosis. Quantitative real-time PCR was used to measure the mRNA expression levels of the key regulatory factors of unfolded protein reaction (UPR). A zebrafish xenograft model was used to study the anti-tumor effect of bortezomib, obatoclax and their combination in vivo.@*RESULTS@#Bortezomib or Bcl-2 inhibitors alone inhibited the cell viability of Jurkat cells, but only obatoclax and bortezomib showed synergistic cytotoxicity and pro-apoptotic effect. Obatoclax, rather than AT-101 and ABT- 199, blocked autophagic flux in the cells evidenced by concomitant accumulation of LC3B-Ⅱ and p62. Both bortezomib and obatoclax alone caused accumulation of polyubiquinated proteins, and their combination showed a synergistic effect, which was consistent with their synergistic cytotoxicity. The dual blockade of proteasome and autophagy by the combination of bortezomib and obatoclax triggered unfolded protein response followed by cell apoptosis. Preventing UPS dysfunction by tauroursodeoxycholic acid (TUDCA) significantly attenuated the cytotoxicity and pro-apoptotic effect of bortezomib in combination with obatoclax. In zebrafish xenograft models, bortezomib combined with obatoclax significantly decreased tumor foci formation.@*CONCLUSIONS@#Bortezomib and obatoclax for dual blockade of protein degradation pathways show synergistic anti-tumor effect in human acute T lymphoblastic leukemia cells.

Effect of matrix metalloproteinase-26 on human glioma angiogenesis / 临床与实验病理学杂志

To investigate the effect of MMP-26 on human glioma angiogenesis and the possible mechanism.Methods The MMP-26 plasmid and empty plasmid pcDNA3.1 were stably transfected into U251 cells to establish a nude mice xenograft model,and then an in vitro human tumor tissue-based three-dimensional angiagenic model.Tissue disks were visually assessed over time to determine the percentage of wells that developed an angiogenic response(I％) and the density and length of neovessel growth were graded at intervals using a semiquantitative visual growth-rating scheme (angiogenic index,AI,0-16scale) in groups of MMP-26 transfected U251 cells (U251-MMP-26),pcDNA3.1 vector-transfected U251 cells (U251-pcDNA3.1) and non-transfected U251 cells (U251).RT-PCR and immunohistochemistry were used to detect the expression of mRNA and protein of MMP-26 and VEGF in groups of U251-MMP-26,U251-pcDNA3.1 and U251.Immunohistochemical localization of CD31 was determined in the endothelial tubes invading the fibrin-thrombin clot matrix.Results Immunohistochemical endothelial cell markers CD31 was positive in the vascular tubes invading the fibrin-thrombin clot matrix,confirming their endothelial origin.The angiogenesis results showed that difference of length of micro capillaries,density of branches,and the area occupied between U251-MMP-26 groups and control groups were significant.The percentage of tumor implants that developed invasion (I％) and the angiogenic index AI in U251-MMP-26 group on day 14 were higher than those of U251-pcDNA3.1 group and U251 group (P ＜ 0.05).The trends of I％ and AI in 14 days were significant compared with those in control groups.The expression of mRNA and protein of MMP-26and VEGF in U251-MMP-26 group was significantly higher in U251-MMP-26 group than those in U251-pcDNA3.1 group and U251 group(P ＜0.01).Conclusion The effect of MMP-26 on promoting glioma angiogenesis may be related to the increased expression of VEGF,which can be used as targets for anti-tumor therapy.

CispIatin inhibits survivaI of human esophageaI squamous carcinoma ceIIs via p53 activation / 中国药科大学学报

To study the mechanisms whereby cisplatin suppresses survival of human esophageal squamous cell carcinoma cells.The cytotoxicity of cisplatin in cisplatin-resistant cell line EC109 /CDDP and its parental cell line EC109 was measured by cell viability assay.Western blotting was used to investigate the protein expression of to-tal p53 and phosphorylated p53 at Ser15.Colony formation assay was employed to evaluate the ability of cells to recover from treatments and form colonies.The results indicated that EC109 /CDDP cells were more resistant to cisplatin-induced cytotoxicity than EC109 cells,with the IC50 values of (20.4 ±4.4)μmol /L and (5.7 ±0.1 )μmol /L,respectively.Although cisplatin did not alter the total protein level of p53,it obviously increased the phosphorylation of p53 at Ser15.Cisplatin inhibited survival of both EC109 /CDDP and EC109.Notably,inhibition of p53 by Pifithrin-αsignificantly promoted recovery of cisplatin-treated EC109 and EC109 /CDDP cells to differ-ent degrees.In this respect,p53 protein was found to be activated in response to cisplatin treatment in both EC109 /CDDP and EC109,which may contribute to the cytotoxic effect of cisplatin.

Cellular response to altered autophagy activity in human fibroblast cells overexpressing Ha-RasV12 / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of oncogenic Ras overexpression on autophagic activity in human fibroblast cells in vitro.</p><p><b>METHODS</b>BJ cells were transfected with H-RasV12 or control vector and treated with chloroquine, small interfering RNA (siRNA) for ATG7, or rapamycin. The cellular responses were analyzed by monitoring the parameters and biomarkers for cell growth, senescence and cell death.</p><p><b>RESULTS</b>In BJ cells overexpressing H-RasV12, chloroquine treatment resulted in more prominent cell senescence and a significantly increased cell death rate. Suppression of ATG7 mediated by siRNA also promoted cell senescence. Rapamycin treatment also caused an increased cell death rate but attenuated senescence in surviving cells. In control BJ cells, the cellular response to chloroquine included senescence and cell death, which occurred slowly. Rapamycin treatment and siRNA suppression of ATG7 had no obvious effect on control BJ cells.</p><p><b>CONCLUSION</b>Stable cellular overexpression of oncogenic Ras causes tightly controlled suppression of the autophagic activity of human fibroblast cells, and such changes produce significant effect on cell senescence and survival.</p>

Optimization of a compound prescription for treating liver fibrosis / 南方医科大学学报

<p><b>OBJECTIVE</b>To optimize a compound prescription for treatment of liver fibrosis with an improved therapeutic effect and low toxicity.</p><p><b>METHODS</b>In rat models of liver fibrosis induced by thioacetamide (TAA), the optimized prescription was screened based on a uniform design with 2-factor 5-level table using Uniform Design 3.0 software and tested using liver content of Hyp as the screening index. To verify the efficacy of the optimized prescription, the rat models of liver fibrosis were randomized into normal control group, model group, colchicine group and optimized prescription group, and the changes of hepatic Hyp content, serum HA, ALT, AST, and ALB levels, and the pathology liver fibrosis were observed after corresponding treatments.</p><p><b>RESULTS</b>The optimized prescription, which contained 70 mg/kg glycyrrhizin and 70 mg/kg matrine, showed a significant therapeutic effect against liver fibrosis in rats (Plt;0.05), and the effect was equivalent to that of colchicine (P>0.05).</p><p><b>CONCLUSION</b>Uniform design is a valuable method in prescription optimization. The optimized compound prescription of matrine and glycyrrhizin has a significant effect in inhibiting liver fibrosis.</p>

Expression of tumor suppressor gene product Rb protein in breast cancer and its clinical significance / 吉林大学学报(医学版)

Objective To study the expression of retinoblastoma(Rb) protein in patients with breast cancer and its significance.Methods The expressions of Rb protein were detected in 52 patients with breast cancer,20 patients with breast fibroadenoma or breast cystic hyperplasia(control group) by S-P immunohistochemical method.(Results The negative) rate of Rb expression in breast cancer(76.91%) was higher than that in control group(15.00%)(P0.05).Of all 46 follow-up patients with breast cancer,the peritoneal recurrence and metastatic rate was 10.53% in 38 cases of the negative expression of Rb protein,and while the rate was 0 in 8 cases of the positive expression of Rb protein,there was significant difference(P

Human bone marrow mesenchymal stem cells differentiate toward endothelial lineage in vitro / 中国病理生理杂志

AIM: To investigate the cytological basis and differentiating conditions of human bone marrow mesenchymal stem cells (hMSCs) differentiated into cells of the endothelial lineage in vitro. METHODS: hMSCs were isolated by density gradient centrifugation and fractionated on a 1 073 g/L Percoll. The combination of VEGF165 and various matrix proteins including fibronectin (FN) and type I collagen (Col) was used to induce hMSCs in vitro. Cells were characterized by immunohistochemistry, cytochemistry, FACS and ultrastructure to identify and detect the differentiated population and markers. RESULTS: hMSCs was positive for KDR. PAS reaction was positive and ultrastructure of hMSCs showed glycogen- pool in ectoplasm. Glycogen reducing or disappear suggested that stem cells have occurred differentiation. Induction of hMSCs resulted in the increase of KDR, β1 integrin and CD34. CONCLUSION: hMSCs were induced to a transit population (TP( )) that differentiated toward the endothelial progenitor cells ( EPC), but not a really EPC. hMSCs pedigree diagram of differentiation was hMSCs→TP→EPC→endothelial cells (Ecs).

Experience of Clinical Pharmaceutical Care / 中国药房

OBJECTIVE:To discuss the contents and importance of clinical pharmaceutical care.METHODS:Our experiences in carrying out clinical pharmaceutical care in our hospital were analyzed through exemplification.RESULTS & CONCLUSION:Clinical pharmaceutical care can help improve the medical quality and reduce medical risks.Pharmacist system should be established in hospital to support the work of clinical pharmacists.

Ganglioside inhibit the antigen-presenting capability of epidermal Langerhans cells / 中国免疫学杂志

Objective:To study whether gangliosides inhibit the antigen-presenting capability of epidermal Langerhans cells.Methods:In the vitro test, the purified Langerhans cells were exposed to increasing concentration of gangliosides for 5 hours at 37℃,then KLH was added and incubated for 2 hours at 37℃.At last we added HDK1 and after 72 hours of coculture, levels of IFN-? in culture supernatants were measured by ELISA. In the vivo test, immunity to the S1509a spindle cell carcinoma was induced by s.c. inoculation at weekly intervals into naive syngeneic(CAF1) for a total of three immunizations. Delayed-type hypersensitivity(DTH) was elicited 1 week after the last immunization by injection a hind footpad with TAA-pulsed Langerhans cells incubated with or without gangliosides. Footpad swelling was assessed at 24 and 48 hours with a spring-loaded engineer′s micrometer.Results:The presence of gangliosides during HDK1 activation reduced the expression of IFN-? in vitro test. Gangliosides suppressed Langerhans cells to elicit DTH against TAA in vivo test.Conclusion:Ganglioside inhibit the antigen-presenting capability of epidermal Langerhans cells.

Human bone marrow mesenchymal stem cells differentiate toward endothelial lineagein vitro / 中国病理生理杂志

AIM:To investigate the cytological basis and differentiating conditions of human bone marrowmes-enchymal stem cells(hMSCs) differentiated into cells of the endothelial lineagein vitro.METHODS:hMSCs were isolatedby density gradient centrifugation and fractionated on a 1 073 g/L Percoll.The combination of VEGF165and various matrixproteins including fibronectin(FN) and typeⅠ collagen(Col) was used to induce hMSCsin vitro.Cells were character-ized by immunohistochemistry,cytochemistry,FACS and ultrastructure to identify and detect the differentiated populationand markers.RESULTS:hMSCs was positive for KDR.PAS reaction was positive and ultrastructure of hMSCs showedglycogen-pool in ectoplasm.Glycogen reducing or disappear suggested that stem cells have occurred differentiation.In-duction of hMSCs resulted in the increase of KDR,?1integrin and CD34.CONCLUSION:hMSCs were induced to atransit population(TP) that differentiated toward the endothelial progenitor cells(EPC),but not a really EPC.hMSCspedigree diagram of differentiation was hMSCs→TP→EPC→endothelial cells(ECs).

Different response of antisense IRAK-2 oligonucleotides to PGI2 release induced byIL-1 and TNF / 中国免疫学杂志

To explore the effects of artisense IRAK-2 oligonucleotide on the prostacyclin (PGI2) synthesis in human umbilicalvein endothelial cells (HUVEC) induced by interleukin-1 (IL-1) and tumor necrosis factor (TNF).Methods:The HUVECs were transfectedwith antisense interleulkin-1 receptor associated kinase-2 oligonucleotide (IRAK-20DN) and stimulated with IL-1 and TNF. The levels of PGI2release were analyzed by competitive ELISA. Results: Pre-transfection with antisense IRAK-20DN could remarkably decrease the levels ofPGI2 synthesis induced by IL-1 in time- and dose-dependent manner, whereas it could not attenuate the one stimulated by TNF. Conclusion:The response of antisense IRAK-2 ODN to IL-1 and TNF-stimulated PGI2 release were different. IRAK-2 plays a key role in the IL-1 signalingevents leading to PGI2 release.