Objective To investigate the effects of ATP1A1 knockdown by RNA interference(RNAi) on proliferation of human U251 glioma stem cells .Methods The human U251 glioma stem cells were infected with lentivirus expressing ATP1A1-shRNA . The mRNA and protein expressions of ATP1A1 in U251 glioma stem cells were detected by RT-qPCR and Western blotting ,re-spectively .The cell cycle and apoptosis were evaluated by flow cytometry .The proliferation of U251 glioma stem cells was deter-mined by MTT assay .Results The expressions of ATP1A1 in U251 glioma stem cells transfected with ATP1A1-shRNA were in-hibited significantly at both mRNA and protein levels ,with an inhibitory rate of 84 .15% for ATP1A1 mRNA and of 55 .33% for ATP1A1 protein respectively .The proliferation of cells was inhibited ,the cell apoptotic rate was significantly increased and the cell cycle was arrested in G1 phase and S phase decreased significantly in ATP1A1-shRNA cells(P<0 .05) .Conclusion RNAi targe-ting ATP1A1 gene could down-regulates the ATP1A1 expression ,induces cell apoptosis ,regulates cell phase redistribution and in-hibits cell proliferation in U 251 glioma stem cells .

AIM:TostudythefunctionofmicroRNA(miR)-19aandmiR-92abyseed-targetinginhibitionin multiple myeloma cells and their signal pathways .METHODS:The experiments were divided into t-antimiR-19a group, t-antimiR-92a group, scramble control group and blank control group .The growth-inhibitory potencies were measured by MTT assay.The ability of cell colony formation was measured by cell colony formation assay .The ability of cell invasion was measured by Transwell experiment .The miR-19a and miR-92a target gene signal pathways were integrated by miRFo-cus software.RESULTS:MTT assay showed that t-antimiR-19a and t-antimiR-92a significantly inhibited the viability of multiple myeloma cells , and the best concentration and time were 0.5μmol/L and 48 h, respectively .The colony number in t-antimiR-19a/92a group was less than that in scramble control group .The transfection with t-antimiR-19a or t-antimiR-92a effectively decreased the cell invasion , as the relative invasion cell number was significantly decreased compared with scramble control group.miR-19a and miR-92a were involved in mTOR signaling, cell cycle and other cancer pathways . CONCLUSION:miR-19a and miR-92a cluster might be a potential target for therapeutic intervention in multiple myelo-ma.

Objective To establish abdominal aortic dissection model in ApoE-/-mice, and to evaluate the ability of 7.0T MR to detect the abdominal aortic artery aneurysms in ApoE-/-mice in vivo. Methods ApoE-/-mice aged 10 months were infused with angiotensin Ⅱ with 14 days Osmotic minipump after 10 weeks of high lipid diet. Two different doses of angiotensin Ⅱ were given to mice, i.e. 1000 ng/(kg·min) and 500 ng/(kg·min), respectively. The contrast group was infused with saline water. The abdominal aortic artery was observed in vivo with MR before and within 14 days infusion. At last, the pathological changes of the abdominal artery were compared with MRI findings. Results After 6 or 7 days higher dose of angiotensin Ⅱ infusion, aortic dissection was seen. MR T2WI showed crescent-shaped high signal in the vessel wall of one side，the pathological study identified the hematoma between media and adventitia. Abdominal aortic dissection aneurysms were also found in the mice 13 or 14 days after lower dose of angiotensin Ⅱ infusion, which were consistent with pathological studies. Besides, the signal of the vessel wall was significantly higher in both T2WI and PDWI sequences. There was excellent agreement between MR and histopathology. 〖WTHZ〗 Conclusion Abdominal aortic dissection aneurysms model can be successfully established with different doses (1000 ng/(kg·min) and 500 ng/(kg·min)) of angiotensin Ⅱ infusion into ApoE-/-mice fed with high lipid diet. High-resolution MR is able to visualize the abdominal aortic dissection aneurysm formation in vivo.

Objective To evaluate the application effect and value of four kinds of neuroendoscopic operation mode in the treat-ment of hydrocephalus .Methods The four kinds of neuroendoscopic operation mode ,including endoscopic third ventriculostomy , septostomy of the septum pellucidum ,cyst ventriculostomy and ventrideperitoneal cavity ,were selected in 58 cases of hydrocephalus according to the indications .The treatment effects were analyzed and the application value was evaluated .Results Among 58 cases of hydrocephalus ,50 cases were significantly improved after operation ,the total effective rate was 86 .21% ,2 cases appeared some complications .The four kinds of operation mode could better treat hydrocephalus .Conclusion Neuroendoscopy is an ideal method in treating hydrocephalus and has the advantages of the slight trauma and fewer complications ,which is a method deserving to be promoted in clinic .

0.05).Conclusion HV-CVVH is effective in the treatment of severe acute pancreatitis.For SAP patients who are suit for the hemofiltration treatment,the treatment of HV-CVVH for 8h and for more than 8h has nearly the same effectiveness.

Objective:To study the HPLC fingerprints of Coptidis Rhizoma- Magnoliae Officmalis Cortex formula granules and the differences of active ingredients in different proportions; To explore the content changes of key components in different proportions of Coptidis Rhizoma- Magnoliae Officmalis Cortex. Methods:HPLC was used to determine the contents of several alkaloids and total phenol of Magnolia officinalis in Coptidis Rhizoma- Magnoliae Officmalis Cortex formula granules and their fingerprints, and the similarity evaluation, cluster analysis and principal component analysis were performed. Results:The similarity of fingerprint of 10 batches of Coptidis Rhizoma- Magnoliae Officmalis Cortex was > 0.950. 17 common peaks were identified, and 6 components were identified. Compared with single medicine, the contents of alkaloids and total phenols in the Coptidis Rhizoma- Magnoliae Officmalis Cortex formula granules were significantly reduced. The contents of multiple alkaloids and total phenols in the Coptidis Rhizoma- Magnoliae Officmalis Cortex formula granules in different proportions were different, and the contents of alkaloids and total phenols were the highest when the proportion of Coptidis Rhizoma- Magnoliae Officmalis Cortex was 2∶1. Conclusion:The contents of main components of Coptidis Rhizoma- Magnoliae Officmalis Cortex formula granules with different proportions are different, which can provide a certain basis for studying the compatibility mechanism of TCM couplet medicines.

Chimeric antigen receptor T (CAR-T) cell therapy achieved advanced progress in the treatment of hematological tumors. However, the application of CAR-T cell therapy for solid tumors still faces many challenges. Competition with tumor cells for metabolic resources in an already nutrient-poor tumor microenvironment is a major contributing cause to CAR-T cell therapy’s low effectiveness. Abnormal metabolic processes are now acknowledged to shape the tumor microenvironment, which is characterized by increased interstitial fluid pressure, low pH level, hypoxia, accumulation of immunosuppressive metabolites, and mitochondrial dysfunction. These factors are important contributors to restriction of T cell proliferation, cytokine release, and suppression of tumor cell-killing ability. This review provides an overview of how different metabolites regulate T cell activity, analyzes the current dilemmas, and proposes key strategies to reestablish the CAR-T cell therapy’s effectiveness through targeting metabolism, with the aim of providing new strategies to surmount the obstacle in the way of solid tumor CAR-T cell treatment.

Objective To evaluate the sensitivity and specificity of enzyme assays,and to provide disease spectrum of lysosomal storage diseases (LSDs).Methods Three thousand three hundred and sixty-four high risk individuals were screened for 24 LSDs at Guangzhou Women and Children's Medical Center between January 2009 and December 2016.Twenty-two kinds of enzyme activities from peripheral blood leucocytes or plasma were measured by using the fluorometry or colorimetry of corresponding artificial substrates,screening for 24 LSDs diseases.Measurement data were represented by (x) ± s,and count data were expressed as a percentage or composition ratio.Results A total of 283 subjects were diagnosed with 18 different kinds of LSDs,and the positive rate of high-risk screening was 8.4％.Among the identified patients,172 cases (60.8％) were mucopolysaccharidosis (MPS),79 cases (27.9％) were sphingolipidoses,18 cases (6.4％) were Pompe diseases,10 cases (3.5％) were affected with mucolipidoses,3 cases (1.1％) were glycoprotein storage diseases,and 1 case(0.4％) was Wolman disease.Of the MPS cases,there were 75 cases of MPS Ⅱ (43.6％),45 cases of MP5 ⅣA (26.2％),24 cases of MPS Ⅵ (14.0％) and 20 cases of MPS Ⅰ (11.6％).Gaucher disease (23/79 cases,29.1％) and metachromatic leukodystrophy (MLD) (21/79 cases,26.6％) were common in sphingolipidoses group.Both the sensitivity and specificity of enzyme assays on peripheral blood leucocytes for LSDs were 100％.Conclusions The most common kinds of LSDs are MPS Ⅱ,MPS Ⅳ A,MPS Ⅵ,Gaucher disease,MLD and Pompe disease.Leukocyte enzymology analysis of high-risk screening LSDs has high sensitivity and specificity.

【Objective】 To develop a novel screening reagent for -D- phenotype preliminary screening based on the difference in RhD antigen expression level of -D- phenotype and normal RhD phenotype. 【Methods】 RhD antigen expression of -D-phenotype and Rh D-- gene carrier were detected by flow cytometry. By adjusting the concentration of polybrene in the screening system, the red blood cells with high RhD antigen expression level agglutinated, and the preliminary screening of the -D-phenotype and its gene carriers was realized. 【Results】 According to the quantitative results of immunofluorescence intensity (MFI) analysis by flow cytometry, the expression level of RhD antigen in -D- phenotype cells (284 360±16 698, n=3) was about 3 times normal RhD positive cells (98 642±35 908, n=9)(P<0.01), while RhD antigen expression level of RhD-- gene carrier (181 109±39 455, n=4) was about 2 times normal RhD positive cells(P<0.01). RhD antigen expression (144 538±227 445, n=7) of the positive cells screened by 15 μL 3% fresh red blood cell suspension and screening system 35 μL (1 μL IgG anti-D, 29 μL polybrene polybrene, and 5 μL low ionic strength solution) was about 1.5 times normal RhD positive cells. 【Conclusion】 The polybrene preliminary screening system, which can be used for high-throughput screening of -D- phenotype, is a reliable technical method for frequency study of this phenotype.

【Objective】 To construct an in-vitro model of erythrocyte antibody-mediated complement activation, and establish quantitative detection methods based on flow cytometry and spectrophotometry, so as to explore the correlation of anti-body titers and complement activation speed, and provide a methodological basis for studying the adverse transfusion reactions of anti-body mediated complement hemolysis. 【Methods】 Mouse monoclonal antibody that recognized human C3b and fluorescent secondary antibody were used to label C3b fragments on erythrocytes, and the deposition of C3b fragments after complement activation was detected by flow cytometry. The absorbance at 540 nm of the supernatant in the complement activation reaction system was measured by spectrophotometry as the amount of hemoglobin released was related to the absorbance. 【Results】 The complement activation system was constructed according to the ratio of 3% red blood cell suspension (mixed for 6 people) 1∶anti-Tj^a 1∶complement 2. The repeatability was good (P value>0.05) as different red blood cell mixtures had been used to repeat the detection reaction system. When using 32×, 64× and 128× dilutions of anti-Tj^a mediated complement activation, the deposition of C3b fragments has been detected by flow cytometry at 30 s, 1 min and 2 min, respectively, and MFI peaked at 5 min, 10 min and 30 min, respectively. No obvious hemolysis has been observed within 1.5 h. 【Conclusion】 In vitro model of anti-Tj^a-mediated complement activation demonstrates the speed of complement activation is related to the concentration of antibody. At a certain antibody concentration, the speed of complement activation has been slowed down, and no obvious hemolysis observed.