OBJECTIVETo observe the expression of phospholipid scramblase 1 (PLSCR1) in matrine (MAT) induced differentiation of all-trans retinoic acid (ATRA) resistant acute promyelocytic leukemia (APL) cells, and to explore its correlation to cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) signal pathway.

METHODSNB4 (an APL cell line sensitive to ATRA) and NB4-R1 (a resistant strain of ATRA) were observed as subjects in this study. Effects of combined treatment of 0.1 mmol/L MAT and 1 [mol/L ATRA on the differentiation of two cell lines were detected using nitroblue tetrazolium (NBT) reduction test and flow cytometry (CD11b). Expressions of PML/RARot and PLSCR1 protein/gene were detected using Western blot and Real-time fluorescence quantitative PCR assay. Meanwhile, H89, PKA antagonist, was used to observe cell differentiation antigen and changes of aforesaid proteins and genes.

RESULTSMAT combined ATRA could significantly elevate positive rates of NBT and CD11 b in NB4-R1 cells, and significantly down-regulate the expression of PML/RARapha-fusion protein/gene (P < 0.05, P < 0.01). ATRA used alone could obviously enhance the expression of PLSCRI in NB4 cells at protein and mRNA levels (P < 0.01). But the expression of PLSCR1 was up-regulated in NB4-R1 cells, but with statistical.difference only at the protein level (P <0. 01). In combination of MAT, PLSCR1 protein expression was further elevated in the two cell lines (P < 0.01). Besides, there was statistical difference in mRNA expressions in NB4-R1 cells (P < 0.05). All these actions could be reversed by treatment of 10 micromol/L H89 (P < 0.05, P < 0.01).

CONCLUSIONMAT combined ATRA could significantly induce the differentiation of NB4-R1 cells, and inhibit the expression of PML/RARalpha fusion gene/protein, which might be associated with up-regulating PLSCR1 expression.

Alkaloids ; Antineoplastic Agents ; Cell Differentiation ; Cell Line, Tumor ; Down-Regulation ; Humans ; Leukemia, Promyelocytic, Acute ; metabolism ; Phospholipid Transfer Proteins ; metabolism ; Quinolizines ; RNA, Messenger ; Signal Transduction ; Tretinoin ; Tumor Cells, Cultured ; Up-Regulation

Homo sapiens longevity assurance homologue 2 (LASS2) is a novel gene isolated from a human liver cDNA library by our laboratory, and it is a human homologue of the yeast longevity assurance gene LAG1 (Saccharomyces cerevisiae longevity assurance gene). According to our previous results, LASS2 could interact with subunit c of vacuolar type H(+)-ATPase (V-ATPase), and the overexpression of LASS2 could inhibit the cell growth of a human hepatocellular carcinoma (HCC) cell line, SMMC-7721. In order to understand the role of the interaction between LASS2 and V-ATPase in HCC cell growth, we transiently transfected plasmid pCMV-HA2-LASS2 into HCCLM3, a HCC cell line without the significant expression of endogenous LASS2. The pH-sensitive fluorescence probes, BCECF and BCECF-AM, were used to measure the intracellular and extracellular H(+) concentrations of HCCLM3 cells respectively. The effect of LASS2 gene on apoptosis was evaluated with Annexin-V/FITC and propidium iodide (PI) by flow cytometry. Western blot was used to detect cytochrome c (Cyt c) in the cytosol and mitochondria, as well as pro-caspase-3 in cytosol. The results showed that the cell growth of LASS2-transfected HCCLM3 cells was significantly inhibited compared with that of the mock control. LASS2 transfection increased intracellular H(+) concentration of HCCLM3 cells, while decreased extracellular H(+) concentration. Moreover, LASS2 transfection significantly enhanced the apoptosis of HCCLM3 cells. In LASS2-transfected cells, the amounts of Cyt c increased in the cytosol, while decreased in the mitochondria. Meanwhile, the expression of pro-caspase-3 in the cytosolic extracts was decreased. These results implicate that LASS2 gene might increase intracellular H(+) of HCC cells via the interaction with V-ATPase, thereby inducing cell apoptosis through mitochondrial pathway.
Apoptosis ; Carcinoma, Hepatocellular ; pathology ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Liver Neoplasms ; pathology ; Membrane Proteins ; metabolism ; RNA, Small Interfering ; Sphingosine N-Acyltransferase ; metabolism ; Transfection ; Tumor Suppressor Proteins ; metabolism ; Vacuolar Proton-Translocating ATPases ; metabolism

OBJECTIVETo study the in vitro effects of low-molecular weight heparin (LMWH) and dexamethasone on the hemolysis of red blood cells from paroxysmal nocturnal hemoglobinuria (PNH) patients.

METHODSBy Ham's test and micro-complement lysis sensitive test (mCLST), the changes of hemolysis of red blood cells from 6 PNH patients were tested by adding different doses of LMWH and dexamethasone into the test mixture. The effects of LMWH and dexamethasone on the coagulation of the tested blood samples were also studied by activated partial thromboplastin time (APTT).

RESULTS(1) Either LMWH or dexamethasone could dose-dependently inhibit the hemolysis of PNH red blood cells, and the effects were synergistic when added together. The same dose of LMWH induced a less than 100% prolongation of APTT. (2) Dexamethasone could inhibit the hemolysis in Ham's test and had different effects on the hemolysis by different adding methods in mCLST. LMWH could inhibit the hemolysis in both Ham's test and mCLST.

CONCLUSIONBoth LMWH and dexamethasone could inhibit the hemolysis of PNH red cells and showed a synergistic effect. The mechanisms of the inhibition of hemolysis were different. Furthermore, a tolerable dose of LMWH induced only a limited prolongation of APTT, which might be useful for controlling acute hemolysis and reducing the dose of dexamethasone.

Anti-Inflammatory Agents ; pharmacology ; Dexamethasone ; pharmacology ; Dose-Response Relationship, Drug ; Erythrocytes ; cytology ; drug effects ; Hemoglobinuria, Paroxysmal ; blood ; Hemolysis ; drug effects ; Heparin, Low-Molecular-Weight ; pharmacology ; Humans ; Partial Thromboplastin Time

OBJECTIVETo develop a coexpression plasmid which expressing envelope protein and nucleoprotein of hepatitis B virus and know of its expressing efficiency.

METHODSThe plasmid coexpressing envelope protein and nucleoprotein of hepatitis B virus under the CMV promoter respectively was constructed by gene recombination. Cellular expression was assessed by ELISA.

RESULTSMultiple cloning site was inserted into expression vector contain hepatitis B virus PreS2-S gene. And expression unit containing hepatitis B virus PreC-C was cloned into it. HBsAg and HBeAg was detected both in the culture supernatant and in the cells.

CONCLUSIONThe coexpressing plasmid was constructed successfully and it can express effectively in vitro. This has provided a basis for further research of the therapeutic HBV DNA vaccine.

Cloning, Molecular ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Hep G2 Cells ; Hepatitis B Core Antigens ; genetics ; metabolism ; Hepatitis B Surface Antigens ; genetics ; metabolism ; Hepatitis B virus ; genetics ; Humans

Objective To understand the treatment circumstance of ST-elevation myocardial Infarction (STEMI) patients at public hospitals in Shenzhen.Methods Directed by Public Hospital Administration at Shenzhen Municipality (PHASM) and led by Chest Pain Treatment Quality Control Center at Shenzhen People's Hospital (CPTQCC-SZ),25 public hospitals in Shenzhen, including 15 PCI-capable hospitals and 10 non-PCI-capable hospitals,we investigated on the overall treatment conditions and the STEMI patient treatment situations from October to December 2015 in these hospitals. A regression analysis was performed between a few factors and the success rate of STEMI treatment was reviewed. Results 383 STEMI cases twere registered between October to December 2015 in the 25 public hospitals in Shenzhen,with 324 case treated in PCI-capable hospitals and 59 cases in non-PCI-capable hospitals. There were statistical differences between the PCI-capable hospitals and non-PCI-capable hospital in fields of total number of senior cardiologists (work year ≥ 3 year),total number of beds in general cardiology beds and number of beds in cccu(all P<0.01). There was no difference in the time of obtaining the first ECG at patient arrival between hospitals(P=0.052).Time for laboratory results availability for troporin was significantly shorter in PCI-capable hospital[(25.0±4.2)min vs.(58.0±2.8)min,P=0.002] .Among the PCI-capable hospitals,the mean D-to-B time was 320 minutes, and mean F-to-B time was 380 minutes. In non-PCI-capable hospitals,D-to-N time ranged from 20 to 350 minutes and F-to-N time ranged from 25 to 380 minutes. Conclusions There are gaps among the overall conditions of the public hospitals in Shenzhen. The overall conditions and chest pain treatment conditions of non-PCI-capable hospitals had bigger gaps with PCI-capable hospitals.

OBJECTIVETo evaluate and compare the value of cardiopulmonary exercise test and conventional pulmonary function tests in the prediction of postoperative cardiopulmonary complications in high risk patients with chest malignant tumors.

METHODSFrom January 2006 to January 2009, 216 consecutive patients with thoracic malignant tumors underwent conventional pulmonary function tests (PFT, spirometry + DLCOsb for diffusion capacity) and cardiopulmonary exercise test (CPET) preoperatively. The correlation of postoperative cardiopulmonary complications with the parameters of PFT and CPET were retrospectively analyzed using Chi-square test, independent sample t-test and logistic regression analysis. The P value < 0.05 was considered as statistically significant.

RESULTSOf the 216 patients, 57 did not receive operation due to advanced stage diseases or poor cardiopulmonary function in most of them. The remaining 159 underwent different modes of operations. Thirty-six patients (22.6%) in this operated group had postoperative cardiopulmonary complications and 10 patients (6.3%) developed operation-related complications. Three patients (1.9%) died of the complications within 30 days postoperatively. The patients were stratified into groups based on V(O(2)) max/pred (≥ 65.0%, < 65.0%); V(O(2)) max×kg(-1)×min(-1) (≥ 20 ml, 15 - 19.9 ml, < 15 ml) and FEV1 (≥ 2.0 L, 1.2 - 1.99 L, < 1.2 L) according to the criteria in reported papers. There was statistically significant difference among these groups in the parameters (P < 0.05), the rates of postoperative cardiopulmonary complications were much higher in the groups with poor cardiopulmonary function (V(O(2)) max/pred < 65.0%; V(O(2)) max×kg(-1)×min(-1) < 15 ml or FEV1 < 1.2 L). It was shown by logistic regression analysis that postoperative cardiopulmonary complications were significantly correlated with age, associated diseases, poor results of PFT or CPET, operation modes and operation-related complications.

CONCLUSIONSFEV1 in spirometry, V(O(2)) max×kg(-1)×min(-1) and V(O(2)) max/pred in cardiopulmonary exercise test can be used to stratify the patients' cardiopulmonary function status and is correlated well with FEV1. V(O(2)) max×kg(-1)×min(-1) is the best parameter among these three parameters to predict the risk of postoperative cardiopulmonary complications in patients with chest malignant tumors and borderline cardiopulmonary function.

Adult ; Aged ; Aged, 80 and over ; Arrhythmias, Cardiac ; etiology ; Exercise Test ; Female ; Humans ; Male ; Middle Aged ; Pneumonectomy ; adverse effects ; Pneumonia ; etiology ; Postoperative Complications ; Predictive Value of Tests ; Respiratory Function Tests ; Respiratory Insufficiency ; etiology ; Retrospective Studies ; Spirometry ; Thoracic Neoplasms ; physiopathology ; surgery ; Young Adult

Objective To compare the clinical effect of endoscopic thyroidectomy and open thyroidectomy in the treat-ment of benign thyroid diseases. Methods A total of 120 patients with benign thyroid diseases were selected from March 2015 to June 2017 in the Third Affiliated Hospital of Xinxiang Medical University. The patients were divided into endoscopic opera-tion group(n = 62)and open operation group(n = 58)according to the treatment methods. The patients in the open operation group were treated with conventional open thyroidectomy,and the patients in the endoscopic operation group were treated with endoscopic thyroidectomy. The operation time,intraoperative bleeding volume,postoperative drainage volume and hospitalization time were recorded in the two groups;and the intraoperative and postoperative complications were observed. The pain of the pa-tients in the two groups was evaluated by visual analogue scale(VAS)at the time points of 6,12 and 24 hours after operation. The cosmetic satisfaction of the patients in the two groups was evaluated by numerical scoring system(NSS)at the time points of 1 and 3 months after operation. Results The operation time and hospitalization time in the endoscopic operation group were significantly shorter than those in the open operation group(P < 0. 05),and the intraoperative bleeding volume and postopera-tive drainage volume in the endoscopic operation group were significantly lower than those in the open operation group(P <0. 05). The VAS score of pain in the endoscopic operation group was significantly lower than that in the open operation group at the time points of 6,12 and 24 hours after operation(P < 0. 05). The NSS score in the endoscopic operation group was sig-nificantly higher than that in the open operation group at the time points of 1 and 3 months after operation(P < 0. 05). The in-cidence of complications in the endoscopic operation group and the open operation group was 12. 90％(8 / 62)and 39. 66％(23 / 58)respectively,the incidence of complications in the endoscopic operation group was significantly lower than that in the open operation group(χ2 = 11. 210,P < 0. 05). Conclusion Endoscopic thyroidectomy is reliable in the treatment of benign thyroid diseases. It has the advantages of small trauma,short operation time,quick recovery,light postoperative pain,small sur-gical scar and fewer complications and so on.

BACKGROUND: Acetabulum anterior column shape is complex, and closed to femoral artery, and femoral nerve. Fixed screws easily went into the acetabulum. Currently, there was less data on quantitative anatomy data of pelvic acetabulum fracture with anterior column plate internal fixation. OBJECTIVE: To compare the pelvis gender differences by analyzing digital anatomical features of normal adult acetabulum anterior column section through digital three-dimensional reconstruction and measurement. METHODS: Totally 30 normal adults (half male and half female) received pelvic CT scans, and data were obtained. Using Materialise Mimics Innovation Suite 16.0 software, the boundary of the pelvis was used as reference line for sectioning. The obturator groove, iliopubic eminence, anterior inferior iliac spine, and the anterior superior iliac spine were used as a reference mark. In 15 males and 15 females (30 sides), the corresponding boundary line from obturator groove to the anterior superior iliac spine of acetabular anterior column was sliced into 5 mm-thick sections. The tangent line was vertical to the boundary line. The tangent plane was vertical to the upper plane of the anterior column. The angle and length of each section 5, 10, 15 mm points from the boundary line to the acetabulum, and the perpendicular distance from anterior and posterior edges of the acetabulum to anterior inferior iliac spine, iliopubic eminence and the pubic tubercle. RESULTS AND CONCLUSION: (1) Acetabular anterior column from the obturator groove to the anterior superior iliac spine section was not significantly different (P > 0.05). (2) No matter in males or females, the tangent angle of the fifth layer section was minimum, and the tangent length of the sixth layer section was longest. The length and angle of the second slice at 5 mm point were not significantly different between males and females. However, above indexes in others were significantly larger in females than in males (P < 0.05). (3) Results indicated that in different fracture ranges and different fixation plate positions, the angle and length of pedicle screws are dynamic, so we only selected in accordance with above range, but cannot fix in a certain value. The design of the most accurate and effective placement angle and length should be aimed at nailing design parameters for each individual patient. The use of Mimics software can be used for three-dimensional reconstruction of CT data of adult acetabular anterior column fracture, and can measure the number of indexes, and provide a theoretical reference for the clinical diagnosis and treatment of acetabular anterior column fracture.

OBJECTIVETo evaluate the long-term outcome of immunosuppressive therapy (IST) in patients with severe aplastic anemia (SAA).

METHODSHematopoietic recovery (peripheral blood cell counts, bone marrow aspirates, bone marrow biopsy, in vitro culture of hematopoietic progenitors), immunity of T lymphocyte, quality of life and side-effects of the therapy were assessed in 50 SAA patients who have survived more than 3 years after IST.

RESULTSAt 3 years, 4 years and 5 years follow-up, 81.5% (13 cases), 86.7% (13 cases) and 89.5% (17 cases) of the SAA patients reached and maintained normal peripheral blood cell counts, 93.4% (15 cases), 93.3% (14 cases) and 94.7% (18 cases) showed normal bone marrow pictures, and 37.5% (6 cases), 40.0% (6 cases) and 73.7% (14 cases) had normal yields of bone marrow cell culture, respectively. Overall, 86.0% (43 cases), 94.0% (47 cases) and 52.0% (26 cases) of the total SAA patients were normalized in peripheral blood counts, bone marrow picture and culture of hematopoietic progenitor yields, respectively. During the follow-up, 88.0% (44 cases) of the patients achieved 100 of Karnofsky scores; 26 of the 31 patients (83.9%) who received bone marrow biopsy showed normal histological pictures, and 29 of 37 patients (78.4%) tested had normal subsets of T lymphocytes. No clonal disease was found. The late side-effects of IST were mild. All of the parameters tested were normal in 24 patients.

CONCLUSIONAfter IST, the hematopoietic function of bone marrow, the immunity of the T lymphocyte and the life quality were normalized with few side-effects in patients with SAA. These patients would probably be cured.

Adolescent ; Adult ; Anemia, Aplastic ; drug therapy ; mortality ; Blood Cell Count ; Bone Marrow Examination ; Child ; Disease-Free Survival ; Female ; Follow-Up Studies ; Hematopoietic Stem Cells ; cytology ; physiology ; Humans ; Immunosuppressive Agents ; therapeutic use ; Karnofsky Performance Status ; standards ; Male ; Middle Aged ; Recovery of Function ; physiology ; Survival Rate ; Treatment Outcome

OBJECTIVETo assess the safety and effects of 40 mg atorvastatin on serum lipids, inflammatory markers and clinical events in ACS patients post PCI.

METHODSA total of 92 patients with ACS post successful PCI were randomly divided into atorvastatin 10 mg/d (group A) and atorvastatin 40 mg/d (group B) on top of the standard medical therapy. Blood were taken at baseline, 4, 12 and 24 weeks for serum alanine aminotransferase (ALT), lipids, high-sensitive C-reactive protein (hs-CRP) and matrix metalloprotease-9 (MMP-9) measurements. The major adverse cardiac events (MACE) were also observed.

RESULTSThere was no significant difference in medication withdrawn (2 vs. 3 cases) due to increased ALT (3 times higher than normal) and incidence of MACE (5 vs. 7 cases) between the groups. TC and LDL were significantly reduced in both groups 4 weeks and thereafter post medication compared to pre-treatment (P < 0.05) and the reduction was more significant in group B than that in group A at 24 weeks post medication (P < 0.05) while TG and HDL remained unchanged. hs-CRP was significantly reduced at 12 and 24 weeks in both groups compared to baseline and the reduction was more significant in group B than that in group A at 24 weeks. MMP-9 was significantly reduced in both groups 4 weeks and thereafter post medication compared to pre-treatment (P < 0.05) and the reduction was more significant in group B than that in group A at 12 weeks post medication (P < 0.05).

CONCLUSIONBoth atorvastatin doses significantly reduced TC, LDL, hs-CRP and MMP-9 in ACS patients post PCI and the reduction was more significant in high dose atorvastatin group at 24 weeks while the MACE and drug withdraw rates were similar between the groups.

Acute Coronary Syndrome ; blood ; drug therapy ; Alanine Transaminase ; blood ; Angioplasty, Balloon, Coronary ; Atorvastatin Calcium ; C-Reactive Protein ; metabolism ; Heptanoic Acids ; therapeutic use ; Humans ; Hypolipidemic Agents ; therapeutic use ; Matrix Metalloproteinase 9 ; blood ; Prospective Studies ; Pyrroles ; therapeutic use