Objective To construct eukaryotic vectors expressing short hairpin RNAs ( shRNAs ) targeting at the CIP2 A gene and to explore its effects on gastric cell line BGC-823 .Methods Four oligonucleotides targeting the CIP2A gene were synthesized and cloned into the eukaryotic expression plasmid pGPU 6.The recombinant plas-mids, pGPU6/GFP/Neo-CIP2A-shRNA-1, 2, 3 and 4, were introduced into BGC-823 cells by lipofectamine-me-diated transfection and the infection rate was observed by fluorescence microscope .The gene silencing efficiency was measured by real-time PCR and Western blot .The effects on proliferation of BGC-823 cells were detected by CCK-8 .Results DNA sequencing and enzyme digestion analysis confirmed the identity of the four recombinant shRNA expression vectors .Immunofluorescsence demonstrated that transfection efficiency was above 70%.Trans-fection of shRNA-1, 2, 3 and 4, significantly knocked down the expression of CIP 2A mRNA and protein at 24, 48 and 72 h after transfection .Compared with the 2 , 3 and 4 , shRNA-1 had the more strong inhibitory effect on the expression of CIP2A mRNA and protein.The CCK-8 assay showed that the anti-proliferation effect on BGC-823 cells was significant ( P<0.05 ) .Conclusions The recombinant vector may effectively inhibit the expression of CIP2A in BGC-823 cells and depress the proliferation of BGC-823 cells.

Objective To observe the application effect of Yunnan baiyao powder and alcohol with VitB12 on exosmosis of vincristin. Methods The 40 patients with exosmosis of vincristin were randomly divided into the treatment group(22 cases)and the control group( 18 cases).The treatment group was given the Yunnan baiyao powder and alcohol with VitB12,the control group was given magnesium sulfate for hydropathic compress.The effective rate and red swelling and ulcer of the skin in the two groups were evaluated. Results The effective rate of the treatment group was higher than that of the control group,red swelling and the diameter of the ulcer of the skin on the fifth and seventh day were shorter than the control group. Conclusions The Yunnan baiyao powder and alcohol with VitB12 are effective in treatment of the exosmosis induced by vincristin.

Background and purpose:Overexpression of inhibitor of protein phosphatase 2 A-2 (I2PP2A) in many tumors including gastric cancer suggests that I2PP2A may contribute to the development of gastric cancer. To further study the biological function of I2PP2A and its role in gastric cancer, we established a BGC823 cell line for stable expression of shRNA targeting human I2PP2A gene. Methods: A double-stranded shRNA targeting the I2PP2A was designed, synthesized and was inserted into a lentivirus vector (pGLV2), and the insertion was identiifed by restriction endonuclease analysis and DNA sequencing. BGC823 cells were then transfected with the packaged recombinant lentivirus, and resistant cell clones were selected with puromycin. The expression of I2PP2A was examined using real-time PCR (RT-PCR) and Western blot. Results:Sequencing result proved that recombinant lentivirus vector pGLV2-shRNA-I2PP2A was constructed correctly. RT-PCR and Western blot results conifrmed that the expression of I2PP2A was signiifcantly down-regulated in this infected BGC823 cell line. The efifciency of siRNA interference of I2PP2A could be up to about 90%. Conclusion:A lentiviral vector carrying a shRNA targeting the I2PP2A gene is successfully constructed, and a BGC823 cell line stably expressing I2PP2A shRNA is established with this lentiviral system.

OBJECTIVETo understand the clinical feature and natural course of polycythemia vera (PV).

METHODSThe clinical symptoms, signs, laboratory examination and prognosis of 185 patients with PV were analysed.

RESULTSThere are 122 males and 63 females. The mean age was (52.7 +/- 14.1) years. The mean hemoglobin level was (208.3 +/- 21.2) g/L. Pancytosis was displayed in 74 (40%) cases, excess of red blood cells in 33 (17.8%), excess of red blood cells and granulocytes in 67 (36.2%) and excess of red blood cell and platelets in 11 (5.9%). Splenomegaly was found in 123 (66.5%) patients and hepatomegaly in 30 (16.2%). Quantitative assess of serum Epo was done in 25 patients. The level was low in 16 (64.2%) and normal in 9 (36.0%). Hematopoietic progenitor culture yields was elevated in 11 patients, endogenous erythroid colonies (EEC) formation was found in 10 cases (90.9%). Eighty two patients (44.3%) had 101 attacks of vascular thrombotic incidents, 7 patients developed myelofibrosis (MF). Secondary cancer occurred in 1 patient. Two patients died of thrombosis.

CONCLUSIONPV is an elderly adult myeloproliferative disease with a high frequency of thrombosis. EEC can be found out in PV patients. The serum Epo level is not increased in PV patients. The main sequelae of PV is MF.

Adult ; Aged ; Erythrocyte Count ; Female ; Hemoglobins ; metabolism ; Hepatomegaly ; etiology ; Humans ; Leukocyte Count ; Male ; Middle Aged ; Polycythemia Vera ; blood ; complications ; pathology ; Primary Myelofibrosis ; etiology ; Splenomegaly ; etiology ; Thrombosis ; etiology

OBJECTIVETo detect the quantity, proportion and function of producing cytokines of Th1 and Th2 cells in aplastic anemia (AA) patients and their contribution to the hematopoietic failure.

METHODS(1) Eleven patients with severe aplastic anemia (SAA) at diagnosis were observed by Marsh's method for the CFU-E, BFU-E and CFU-GM before and after depletion of CD(4)(+) T lymphocytes from bone marrow mononuclear cells (BMMNC); (2) Th1 (CD(4)(+) IFN-gamma(+)) and Th2 (CD(4)(+) IL-4(+)) cells in peripheral blood mononuclear cells (PBMNC) of 21 SAA patients and 17 normal controls were counted by FACS. (3) mRNA expression of IFN-gamma and IL-4 gene in unstimulated BMMNC from 16 SAA patients, 11 chronic aplastic anemia (CAA) patients, 26 other hematological diseases patients and 11 normal controls were measured by reverse transcriptase polymerase chain reaction (RT-PCR).

RESULT(1) CFU-E, CFU-GM and BFU-E increased significantly after depletion of CD(4)(+) T lymphocytes from BMMNC of SAA patients. (2) The percentage of IFN-gamma producing CD(4)(+) T cell (Th1) of SAA patients was significantly higher than that of controls, the percentages of IL-4 producing CD(4)(+) T cells (Th2) had no difference between SAA patients and normal controls. (3) IFN-gamma mRNA was detected in unstimulated BMMNC in 13 of 16 SAA patients, 6 of 11 CAA patients and one of 6 paroxysmal nocturnal hemoglobinuria (PNH) patients. The IFN-gamma mRNA was not detected in unstimulated BMMNC of 11 normal controls and other hematological diseases patients.

CONCLUSIONSDisbalance of CD(4)(+) T lymphocytes subsets and increases in quantity and IFN-gamma producing function of Th1 cells might be important for the development of bone marrow failure in AA and in distinguishing AA from other kinds of pancytopenic diseases.

Adolescent ; Adult ; Anemia, Aplastic ; blood ; etiology ; Colony-Forming Units Assay ; Erythroid Precursor Cells ; cytology ; Female ; Granulocytes ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans ; Interferon-gamma ; genetics ; Interleukin-4 ; genetics ; Macrophages ; cytology ; Male ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Th1 Cells ; cytology ; metabolism ; physiology ; Th2 Cells ; cytology ; metabolism

OBJECTIVETo examine the quantity and apoptosis-related protein level of B lymphocyte in the patients with immunorelated pancytopenia (IRP) and explore the role of B lymphocyte in the pathogenetic mechanism of IRP.

METHODSQuantities of all B lymphocytes and CD(5)(+) B lymphocytes and the expressions of Fas and bcl-2 on B lymphocytes in 25 patients with untreated IRP, 15 IRP patients in complete remission (CR) and 10 normal controls were assayed by FACS.

RESULTSThe percentages of B lymphocyte and CD(5)(+) B lymphocytes were significantly higher in untreated IRP patients than in CR IRP patients and normal controls (P < 0.05); there was no significant difference between the latter two groups (P > 0.05). There was no significant difference of Fas expression in B lymphocytes among the three groups (P > 0.05). The expression of bcl-2 on B lymphocytes was significantly higher in untreated patients than in CR patients or normal controls (P < 0.05), and so did in CR patients than in normal controls (P < 0.01). The apoptosis-related index was significantly lower in untreated patients than in CR patients or normal controls (P < 0.01), and was lower in CR patients than in normal controls (P < 0.05). The percentage of B lymphocyte was positively correlated with the duration from the beginning of treatment to response.

CONCLUSIONThe production of auto-antibodies in IRP patients probably has some relationships with the abnormal quantities of B lymphocyte and its subsets, and with the inhibition of B lymphocyte apoptosis.

Adolescent ; Adult ; Apoptosis ; physiology ; B-Lymphocytes ; classification ; immunology ; pathology ; Bone Marrow ; physiopathology ; CD5 Antigens ; immunology ; Cell Count ; Child ; Female ; Flow Cytometry ; Humans ; Immune System Diseases ; immunology ; metabolism ; physiopathology ; Male ; Middle Aged ; Pancytopenia ; immunology ; metabolism ; physiopathology ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; metabolism ; fas Receptor ; analysis ; immunology ; metabolism

OBJECTIVE: To detect chromosomal aberrations by using cytoplasmic light chain immunofluorescence with fluorescence in situ hybridization (cIg-FISH), and to explore the correlation of del(17p13) with clinical characteristics, drug response and prognosis among patients with newly diagnosed multiple myeloma (NDMM). METHODS: Clinical data of 198 cases of NDMM was collected. cIg-FISH and a specific probe (TP53) were used to detect karyotypic abnormalities in bone marrow samples derived from the patients. Correlation between karyotypic abnormalities and clinical data was analyzed. RESULTS: Nineteen of the 198 patients (9.6%) were found to have a karyotype involving del(17p13). The overall survival (OS) and progression-free survival (PFS) for patients with or without del(17p13) was significantly different (P<0.01). No significant difference was found in OS and PFS between patients carrying a del(17p13) on bortezomib and non-bortezomib regimen (OS: P = 0.873; PFS: P = 0.610). CONCLUSION cIg-FISH is a simple and convenient method for the detection of karyotypic anomalies in multiple myeloma. Del(17p13) is an indicator for poor prognosis for multiple myeloma patients. Bortezomib cannot improve the survival disadvantage of del(17p13).

Objective To investigate the risk factors for acute ischemic stroke in patients with lower extremity atherosclerosis (LEA).Methods The consecutive patients with acute ischemic stroke admitted to hospital within 7 d after onset were enrolled retrospectively.Color Doppler flow imaging was used to detect LEA.The demographic characteristics,vascular risk factors,and laboratory parameters were identified and analyzed.Results A total of 156 patients with acute ischemic stroke were enrolled,including 138 with LEA.Univariate analysis showed that age (69.5± 11.8 years vs.60.4± 11.5 years;t =3.063,P =0.003) and the proportion of patients with hypertension (81.1％ vs,55.6％;x2 =2.467,P =0.014) in the LEA group were significantly higher than those in the non-LEA group.Multivariate logistic regression analysis showed that after adjustment for confounders such as gender,baseline systolic blood pressure,diabetes mellitus,and ischemic heart disease,age (odds ratio [OR] 1.059,95％ confidence interval [CI] 1.016-1.105;P=0.007),and hypertension (OR 3.128,95％ CI 1.084-9.026,P =0.035) were the independent risk factors for acute ischemic stroke complicated with LEA.Conclusions Age and hypertension are associated with acute ischemic stroke complicated with LEA.

Objective To investigatethe significance of serum carbohydrate antigen (CA) 125,car cinoembryonic antigen (CEA),cytokeratin 19 fragment (CYFRA21-1),neuron-specific enolase (NSE),and squamous cell carcinoma antigen (SCCA) in the diagnosis of bone metastasis from lung cancer.Methods A total of 222 patients,including 91 lung cancer patients with bone metastasis (49 males,42 females,average age (60.07± 10.60) years;group A),75 lung cancerpatientswithout bone metastasis (57 males,18 females,average age (62.20± 12.63) years;group B),56 patients with benign lung diseases (34 males,22 females,average age (61.45± 10.66) years;group C) were recruited from January 2015 to January 2016.The electrochemiluminescence was applied to detect serum levels of CA125,CEA,CYFRA21-1,NSE and SCCA.Kruskal-Wallis,Wilcoxon rank sum test,x2 test and receiver operating characteristic (ROC) curve analysis were used to analyze data.Results The levels of serum CA125,CEA,CYFRA21-1,NSE in group A were higher than those in group B and C (H values:13.45-44.96,all P＜0.05);while SCCA was not significantly different among the 3 groups (H=2.56,P＞0.05).The areas under ROC curves for CA125,CEA,CYFRA21-1,NSE,SCCA were 0.667,0.702,0.602,0.664,0.440,respectively.The positive rate of serum NSE in small cell lung cancer (SCLC) patients was higher than that in non-small-cell lung cancer (NSCLC) patients (17/18 vs 32.88％ (24/73);x2=22.11,P＜0.05);CEA was highly expressed in adenocarcinoma,and SCCA was highly expressed in squamous cell carcinoma.Patients with grade Ⅲ+ Ⅳ metas tasis (n =52) had higher levels of CA 125,CEA,NSE compared to patients with Ⅰ + Ⅱ metastasis (n =39;z values:from-2.54 to-0.32,all P＜0.05).The sensitivity of combined detection of 5 tumor markers was 97.80％ (89/91),which was significantly higher than that of single tumor marker (x2 values:35.46-138.23,all P＜0.05).Conclusion Serum levels of CA125,CEA,CYFRA21-1,NSE and SCCA play a role in the diagnosis of bone metastasis from lung cancer,and the combined detection of the 5 tumor markers contribute to the early detection of bone metastases.

Objective Using Z-score to assess the prevalence of proximal aorta dilatation in middle-aged and aged individuals during routine transthoracic echocardiogram examinations and to identify its risk factors. Methods A total of 823 middle-aged or elderly patients on routine transthoracic echocardiogram examinations were consecutively enrolled. The internal diameters of the sinus of Valsalva (SoV ) and the ascending aorta (AA ) were measured. Z-scores were calculated according to the proposed equation for SoV and AA. A dilated aortic root was defined as a Z-score ≥1.96 or the diameter of SoV or AA ≥ 40 mm. The prevalence of proximal aorta dilatation and associated factors were analyzed. Results The prevalences of proximal aorta dilatation ,SoV dilatation ,and AA dilatation were 26.1%(25/823 ) ,6.0%(49/823 ) ,and 23.7%(195/823 ) , respectively.In the aortic root dilatation group ,age and the proportion of obesity were higher (both P＜0.05) ,and there were more female subjects (30.5% or 117/384 vs.22.3% or 98/439 ,P＜0.01) . The incidences of left atrial dilation ,left ventricular dilation ,left ventricular hypertrophy ,and aortic regurgitation in the aortic root dilatation group were higher than those in the non-aortic root dilatation group(P＜0.05 ) .Logistic regression analysis demonstrated that sex (OR＝ 1.827 ,95% CI :1.248-2.673 ,P＝0.002) ,hypertension (OR＝1.441 ,95% CI :1.000-2.075 ,P＝0.050)and left ventricular hypertrophy (OR＝1.827 ,95% CI :1.248-2.673 ,P＝0.002)were independently correlated with aortic root dilatation. Conclusions The prevalence of proximal aorta dilatation is high in middle-aged and aged individuals. Proximal aorta dilatation is related to sex ,age ,and body size ,and it is often accompanied by structural abnormalities of the heart.