Acute Disease ; Child ; Gene Expression Profiling ; Humans ; Leukemia ; diagnosis ; genetics ; therapy

AIM:To differentiate bone marrow mesenchymal stem cells(BMSCs) into functional insulin-producing cells to produce sufficient pancreatic islet cells for transplantation.METHODS:Recombinant adenovirus vectors carrying PDX1 and NKX6.1 genes were constructed and the bone marrow mesenchymal stem cells were infected by the recombinant adenovirus combined with several cytokines for differentiation.The expressions of PDX1,NKX6.1 and insulin and C-peptide in the differentiated bone marrow mesenchymal stem cells were detected by RT-PCR and Western blotting.After the differentiated bone marrow mesenchymal stem cells were transplanted into subrenal capsule of diabetic mice,cell morphology of the grafts as well as their secretion of insulin and C-peptide were detected.Besides,regulating capacities of grafts on the blood glucose level of the diabetic mice were also detected.RESULTS:BMSCs induced by recombinant adenovirus(pAdxsi-CMV-PDX1/CMV-NKX6.1) and several cytokines showed positive dithizone staining and the expressions of ?-cell related molecule such as insulin and glucose transporter 2 were detected by RT-PCR,which showed a sustaining and stable expression.The similar results were showed by Western blotting,immunohistochemical staining and indirect immunofluorescence.The insulin secretions in the cells stimulated with glucose at concentrations of 5.5 and 25 mmol/L in the experimental group were(1 240.4?109.3) mU/L and(3 539.8?245.1) mU/L,respectively,and were significantly higher than those in control group.Moreover,transplantation of the cells to STZ mice in treatment group made serum glucose recover to normal level.CONCLUSION:PDX1 and NKX6.1 gene-modified bone marrow mesenchymal stem cells differentiate into insulin-producing cells in vitro.When these cells transplanted into STZ induced diabetic mice,their serum glucose could return to the normal level and they could live well.Thus this is a promising method for diabetes treatment.

To explore the expressional information of PIWIL4 in human ovarian cancer, semi-quantitative reverse transcription polymerase chain reaction(semi-qRT-PCR) was used to detect the mRNA expression level of PIWIL1, PIWIL2, PIWIL3 and PIWIL4 in human ovarian clear cell carcinoma ES-2 cells.Meanwhile, in human ovarian cancer and adjacent normal tissues, the mRNA expression of PIWIL4 was determined.Then, PIWIL4-siRNA, which was designed to target PIWIL4 and chemical synthesized, was transfected into ES-2 cells with lipofectamine.MTT assay and clone formation assay were used to investigate the effects of PIWIL4-siRNA on the cell growth activity and proliferation capacity of ES-2 cells.Experimental results showed that the mRNA expression level of PIWIL4 was the highest compared to PIWIL1, PIWIL2 and PIWIL3 in ES-2 cells(P

AIM: To construct a recombinant plasmid vector containing human pancreatic duodenal homeobox1(Pdx1) and neurogenic differentiation 1(NeuroD1) genes,and to detect its effective expression in eukaryocytes and the ability to induce differentiation of hepatocytes.METHODS: Using human embryo pancreas mRNA as template,Pdx1 and NeuroD1 genes were amplified by RT-PCR and cloned into the two different multiple cloning sites(MCSA and MCSB) of plasmid pIRES.The recombinant plasmid pI/Pdx1/NeuroD1 was transfected into L02 cells.The expression of Pdx1 and NeuroD1 in transfected cells was detected by immunocytochemistry,IFA,RT-PCR and Western blotting,respectively.RESULTS: The length and sequence of cloned segments were correct.Pdx1 and NeuroD1 were expressed in eukaryocytes.Furthermore,the hepatic cells were induced to express glucose transporter 2(GLUT2) and eukaryocyte transcription factor Nkx6.1,which were functionally correlated to ? cells.CONCLUSION: pI/Pdx1/NeuroD1 plasmid is successfully constructed and expressed in human eukaryocytes,with which the cells express the eukaryocyte transcription factor and GLUT2,indicating the transfected cells functionally correlates to ? cells.The results suggest that Pdx1 and NeuroD1 genes can induce the differentiation the cells from hepatic cells to pancreatic endocrine secretion cells.

Objective: To compare the clinical efficacy and safety between tocilizumab (TCZ) combined with methotrexate (MTX) and TCZ used alone in the patients with active rheumatoid arthritis (RA) by meta-analysis to provide reference for the treatment adjustment for the patients with poor response to MTX.Methods: Literature searches were conducted in sinoMed, VIP, PubMed, Cochrane Library and OVID, and then identified according to the inclusion and exclusion criteria.Meta-analysis was conducted using Rev Man 5.3.5 software.Results: Totally 7 randomized controlled trials (RCTs) were included in the analysis.A total number of patients were 1534 cases.The meta-analysis showed that in the treatment of RA, TCZ combined with MTX had significantly higher odds of DAS28≤2.6 remission and CDAI≤2.8 remission when compared with TCZ alone , and the incidence of adverse events (AE), serious adverse events (SAE) and serious inection had no statistical differences.Conclusion: The efficacy of TCZ combined with MTX is higher than that of TCZ alone in RA patients, and the safety is similar.The conclusion still needs further confirmation by RCTs with large samples and high quality.

Objective To analyze the application of quality control cycle (QCC) in reducing the false negative rate of minimal residual disease (MRD) of flow cytometry in patients with acute myeloid leukemia (AML). Methods In AML patients with abnormal fusion gene detected in hematology laboratory of Changhai Hospital during the year of 2014, the prevalence of AML-MRD detected both by flow cytometry (FCM) and real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) were analyzed retrospectively. The possible causes of false negative rate of flow cytometric MRD referring to PCR were further deeply analyzed, and the improvement measures were adapted from January 2015 to December 2015 and further judged all according to the QCC methods. Results Pareto diagram showed that the dilution and coagulation of the specimen, the improper analysis strategy and the incomplete combination of the MRD index [composition ratio:83.3 % (60/72)] were the main factors leading to the leakage of FCM MRD in 2014. The QCC group devised measures to reduce the dilution probability of bone marrow and develop a standard operating procedures (SOP) for sampling and testing, strengthen the maintenance of the flow instrument and more importantly, focused on optimizing the antibody panels and gated strategies referring to the current two main kinds of MRD detection combination modes on the basis of the latest advances published in 2015. Finally, the undetected rate of AML-MRD was reduced by FCM from 14.8 % (72/486) in 2014 to 2.6 % (16/620) in 2015. Conclusions The QCC can effectively reduce the leakage rate of flow cytometric AML MRD, improve the ability of laboratory quality control and the ability to solve problems. Solving problems with QCC is thus worthy of being popularized.

AIM: To investigate the changes of the L-type calcium channel subunit expression and calcium currents (I_~Ca -L) in cultured rat ventricular myocytes infected by coxsackie virus B_3 (CVB_3). METHODS: Primary cultured neonatal rat ventricular myocytes were infected with CVB_3. The changes of L-type calcium channel subunits mRNA in normal and infected myocytes were measured by semi-quantitative reverse transcription-polymerase chain reaction. I_~Ca -L was recorded in two groups respectively using whole cell patch-clamp techniques. RESULTS: The expression of ?_1 and ? subunits of L-type calcium channel mRNA increased in the infected group compared with the normal one (4.00?0.07 vs 2.21?0.41, P0.05). The average current density of I_~Ca -L significantly increased by CVB_3 infection [(-8.66?0.99) pA/pF vs (-6.97?1.75) pA/pF, P

AIM: To determine the association between subfoveal choroidal thickness before therapy and therapeutic activity in diabetic macular edema.
METHODS: The current study was a retrospective study, which included 32 patients ( 32 eyes ) diagnosed with diabetic retinopathy and macular edema. All the patients were firstly treated with intravitreal injections of ranibizumab. Main outcome measures were included the subfoveal choroidal thickness, central macular thickness and best-corrected visual acuity ( BCVA) at preoperation and postoperative visit at 3mo.
RESULTS: After 3 monthly intravitreal injections of ranibizumab, the BCVA was significantly higher than that before therapy and accompanied with significantly reduced thickness of subfoveal choroid and central fovea of macula. Spearman analysis was revealed that a greater baseline subfoveal choroidal thickness was associated with a better BCVA (rs=0. 544, P=0. 036).
CONCLUSION:In the therapy of intravitreal injections of ranibizumab on diabetic macular edema, there seems to be a better BCVA in the patients with a greater baseline subfoveal choroidal thickness. Therefore, baseline subfoveal choroidal thickness may be a useful predictor for the therapy of diabetic macular edema.

AIM:To assess changes of tear film function in patients after pterygium excision combined with amniotic membrane transplantation.
METHODS:Totally 126 patients with pterygium excision with amniotic membrane transplantation from January 2011 to November 2013 were entered in the study. The tear breakup time ( BUT) , the Schirmer I test ( SⅠt) and tear ferning test ( TFT ) were elevated in the patients before and after pterygium excision combined with amniotic membrane transplantation. The examnation times were 1d before surgey, 1wk, 1, 2mo after surgery. Operation eyes were studied group, while opposite healthy eyes as control group.
RESULTS: Compared with the control group, BUT and TFT were significantly different in the eyes with pterygium (P<0. 05); However, no obvious difference was detected in the results of SⅠt (P>0. 05). The results of BUT and TFT at 1mo after surgery in study group were significantly better than 1wk (P<0. 05), while no significant difference compared with 2mo (P>0. 05); The tear film stability in the study group at 1wk after surgery was still inferior to the control group (P<0. 05) and there was no significant difference at 1, 2mo after surgery (P all>0. 05). SⅠt results did not differ between the different examination times(P>0. 05).
CONCLUSION:Tear film stability was broken in the eyes with pterygium. Pterygium excision combined with amniotic membrane transplantation can obviously restore the tear film function into normal state, and the tear film function could reach steady-state 1mo after surgery.

OBJECTIVETo observe the efficacy of Zaozhu Yinchen Recipe (ZZYCR) on non-alcoholic steatohepatitis (NASH) patients, and to explore its effect on serum free fatty acid (FFA) and tumor necrosis factor alpha (TNF-alpha).

METHODSTotally 120 patients with NASH were randomly assigned to the treatment group (60 cases, treated with ZZYCR, one dose per day) and the control group (60 cases, treated with Silibin Meglumine Tablets, 20 mg each time, thrice per day). The therapeutic course for all was 24 weeks. Serum levels of ALT and AST activities, TC and TG levels were detected before and after treatment. Peritoneal CT was performed in all patients, and CT ratios of liver and spleen calculated. NAFLD activity score (NAS) and degree of hepatic fibrosis were assessed using pathological examinations of liver tissue, and efficacy also evaluated. Serum contents of FFA and TNF-alpha were also detected.

RESULTSCompared with before treatment in the same group, activities of ALT and AST, serum levels of TC, TG, FFA, and TNF-alpha, NAS, scores of symptoms and signs all obviously decreased, degree of hepatic fibrosis was obviously improved in the two groups (P < 0.05, P < 0.01). These changes were more obviously seen in the treatment group (P < 0.05). After 24-week treatment, the total effective rate and total clinical efficacy were 80.00% (48/60 cases) and 85.00% (51/60 cases) in the treatment group, obviously higher than those in the control group [60.00% (36/60 cases) and 73.33% (44/60 cases) respectively], with significant difference (P < 0.05, P < 0.01).

CONCLUSIONZZYCR could improve the clinical efficacy of NASH patients, and its mechanism might be associated with inhibiting serum levels of FFA and TNF-alpha.