Objective To explore the variation of E-index and M-index of rabbits with acute pulmonary embolism (APE) under the status of pulmonary hypertension (PHT). Methods Rabbit models of APE with PHT were established. A series of parameters [including peak early diastolic mitral inflow velocity (EM), peak late diastolic mitral inflow velocity (AM) and so on] were obtained with routine echocardiography and tissue Doppler imaging (TDI);and then E-index, M-index were calculated. The parameters before and after APE were compared. Results Twenty-three rabbit models with APE were successfully established, but 3 with atrial fibrillation were excluded. After APE, pulmonary artery pressure increased significantly, EM decreased observably, whereas right ventricular myocardial performance index (RVMPI) increased more evidently than left ventricular myocardial performance index (LVMPI) (P<0.01) did and E-index decreased and M-index increased remarkably. Conclusion Changes of E-index and M-index may provide reference for quantitative assessment of early APE.

Objective To construct the eukaryotic expression vector of pprI gene from Deinococcus radiodurans R1 and investigate its radioresistant effects in eukaryotic cells.Methods A recombinant vector pEGFP-c1-pprI was constructed by DNA recombinant technique.The empty vector pEGFP-c1 and the pEGFP-c1-pprI were transferred into human lung epithelial cells Beas-2B by LipofectamineTM 2000,respectively.Then the infected cells were screened in order to develop a cell line with stable expression of pprI gene.Cell survival rate was tested by clone-forming assay.Cell cycle distribution and apoptosis were detected by a flow cytometry.The fluorescence intensity of reactive oxygen species (ROS) was observed by a fluorescent microscope.γ-H2AX foci in the irradiated cell was detected by immunofluorescence.Results The eukaryotic expression plasmid of pprI prokaryotic gene was constructed and PprI fusion protein was expressed in human lung epithelial cells successfully,and the cell line (2BG) with a stable pprI gene expression was established.After irradiation,the cell survival fraction of 2BG cells was significantly higher than Beas-2B cells so that the value of D0 、Dq and N of the survival curve were increased.Moreover,the fluorescence intensity of ROS and the number of γ-H2AX foci in 2BG cells were also lower than those of B eas-2B cells(F =16.73,19.47,6.94,P ＜ 0.05).Between these two cell lines,the apoptosis rate and cell cycle G2 arrest also had significant difference (F =139.73,237.92,P ＜ 0.05).Conclusions The pprI gene from Deinococcus radiodurans RI can be stably expressed in the eukaryotic cells and it allows the transferred cells to have a radioresistant function.

Objective To observe the immunological effect of tumor cell vaccines to mouse hepatoma A(HepA) treated by Haematoxylin. Methods HepA cell was treated by 0.1% Haematoxylin and made into three vaccines: HepA vaccine with complete Freund' s adjuvant, HepA vaccine with incomplete Freund' s adjuvant; and HepA vaccine without any adjuvants. Five times of immunization were given the grouped mice with the above three vaccines, then active HepA cells (1×105 for each mouse) were inoculated by intraperitoneal injection to attack them; reckoning the mean survival time (MST) of the grouped mice, comparing the immunoprotective action of the three vaccines to the tmnor-bearing mice. Those mice only receiving normal saline (equal volume to the vaccine) were as a control group. Results MST of control group was (23.30±1.24) day; MST of mice receiving H22 vaccine with complete adjuvant was (43.90±15.20) day (P<0.02); MST of mice receiving H22 vaccine with incomplete adjuvant was (39.60±13.77) day (P<0.05); and MST of mice receiving HepA vaccine without any adjuvant was (38.40±12.54) day (P<0.05); As compared with the control group, the three treated groups showed a life-lengthening rate (LLR) 88.41%, 69.96 % and 64.81% respectively. Conclusion The vaccines treated by Haematoxylin give a marked immunoprotective action to those tumor-bearing mice. The HepA vaccine' s immunological effect is increased by the Freund' s adjuvant (complete or incomplete).

Objective To observe the effects of modifiedDanshen Decoction on spermidine/spermine acetyltransferase (SSAT) /polyamine pathways of SD rats with IRI; To investigate its protective mechanism. Methods The model of IRI was established by ligating left anterior descending coronary artery for 30 min followed by reperfusion for 90 min. The SD rats were randomly divided into the control group, sham-operation group, model group and modifiedDanshen Decoction group, with 10 rats in each group. The myocardial infarction size was measured by using TTC staining. The contents of SSAT were measured by ELISA. The SSAT mRNA and SSAT protein expression level were detected with real-time fluorescent quantitative PCR method and Western blot, respectively. The contents of polyamines (putrescine, spermidine, spermine) in cardiac tissue were detected by HPLC. Results Compared with sham-operation group, the myocardial infarction size, the SSAT content, the SSAT mRNA and SSAT protein expression levels of model group increased significantly, the contents of polyamines decreased significantly, with statistical significance (P<0.01); Compared with model group, the myocardial infarction size of modifiedDanshen Decoction group was significantly reduced, while the SSAT content and SSAT mRNA and protein expression level decreased significantly, the contents of polyamines increased, with statistical significance (P<0.05, P<0.01).ConclusionModifiedDanshen Decoction can adjust the SSAT polyamine pathways and increase polyamine content in cardiomyocytes, and thus play a role of protection of myocardial ischemia-reperfusion injury.

Objective To study the distribution and properly of the transparent globules within Hensen cells (HC) of guinea -pig Corti organ .Methods The cochlear epithelial cells were isolated from 10 guinea pigs .The cells of cochlea were marked by Bodipy493/503 ,sudan III ,oil red O ,and osmium tetroxide .Results The transpar‐ent globules within the HCs of the guinea -pigs were green staining by Bodipy493/503 ,jacinth staining by Sudan III ,ruby red by oil red O .And they were black globules stripe as post -fixed in 1% osmium tetroxide .Conclusion The results indicate that the transparent globules within guinea -pigs HCs'lipid droplets by four methods .

Familial isolated pituitary adenoma (FIPA) is an autosomal dominant disease, characterized by low penetrance, early-onset disease, more invasive tumor growth, as well as somatotroph and lactotroph adenomas in most cases. It has been indicated that the aryl hydrocarbon receptor interacting protein (AIP) gene is a tumor suppressor gene. Many heterozygous mutations have been discovered in AIP in about 20% of FIPA families. However, the exact molecular mechanism by which its disfunction promotes tumorigenesis of pituitary is unclear.
Growth Hormone-Secreting Pituitary Adenoma ; genetics ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; Mutation ; Pituitary Neoplasms ; genetics

Objective: To learn the epidemiological characteristics of fall mortality among the elderly people in Taizhou，and to provide basis for intervention strategies of fall in the elderly. Methods: Data of fall mortality among residents aged 60 years or over in Taizhou from 2016 to 2018，collected from Zhejiang Chronic Disease Surveillance Information Management System，was used for analysis of time，population and geographical characteristics of fall deaths. The epidemic trend of fall mortality in the elderly was described by annual percentage change（APC）. Results: From 2016 to 2018，3 699 cases of fall death in Taizhou were reported，the crude and standardized mortality were 116.90/100 000 and 97.88/100 000. The standardized mortality of fall in women was 106.11/100 000，which was higher than 90.13/100 000 in men（P<0.05）. The standardized mortality of fall in rural residents was 131.20/100 000，which was higher than 28.15/100 000 in urban residents（P<0.05）. The mortality of fall in residents aged 65-69 years from 2016 to 2018 showed an upward trend（APC=4.20%，P<0.05），while the mortality trend of fall in other age groups was not statistically significant（P>0.05）. Conclusion Fall was the first cause of injury death in the elderly aged 60 years or over in Taizhou. Females and rural residents have relatively higher fall mortality.

OBJECTIVETo investigate relationship between glucose metabolic disorders and expression of insulin receptor (IR) and tyrosine protein kinase (TPK) in posthepatitic cirrhosis hepatocyte and HBV DNA expression in pancreatic cells.

METHODSTo detect HBV DNA in paraffin-embedded pancreatic and hepatic tissues from 12 posthepatitic cirrhosis patients with positive serum HBV markers by using in situ hybridization (ISH) with a digoxigenin labelled probe. The amount of IR and TPK have been evaluated by immunohistochemical quantitative analysis using image analyzer in hepatocyte of 12 patients positive for HBV markers with posthepatitic cirrhosis in serum. Immunofluorescent histochemical double staining technique was used. HBsAg and IR were observed under confocal laser scanning microscope.

RESULTSEleven of 12 cirrhosis patients? hepatocytes were HBV DNA positive, including 7 patients (7/7) with impaired glucose tolerance (IGT) and 4 patients (4/5) with normal glucose tolerance (NGT). Eight of 12 pancreatic cells were HBV DNA positive, including 7 patients (7/7) with IGT, but only one patient (1/5) with NGT-HBV DNA was found positive in pancreatic cells in significantly more subjects in IGT group than in NGT group (P less than 0.01).IR and TPK amount in hepatocyte of IGT was significantly less than that of NGT patients with posthepatitic cirrhosis (P less than 0.01). IR amount was closely related to the TPK in cirrhosis hepatocyte r=0.82597(P less than 0.01). HBV DNA was mainly localized in the nuclei of hepatocyte and pancreatic acinar and islet cells. Immunofluorescent histochemical double-staining showed that HBsAg was partly localized in the IR positive areas of hepatocytes and pancreatic islet cells.

CONCLUSIONHBV can invade acinar cells of pancreas and islet cells, which might be a direct cause of insulin-dependent diabetes mellitus-like the disorder and insulin absence after HBV infection. Decrease of IR and TPK might be main cause of noninsulin-dependent diabetes mellitus-like disorder after having hepatitis or posthepatitic cirrhosis.

DNA, Viral ; analysis ; Female ; Glucose Metabolism Disorders ; complications ; metabolism ; virology ; Hepatitis B virus ; genetics ; Hepatocytes ; metabolism ; virology ; Humans ; In Situ Hybridization ; Liver Cirrhosis ; complications ; metabolism ; virology ; Male ; Middle Aged ; Pancreas ; cytology ; virology ; Protein-Tyrosine Kinases ; metabolism ; Receptor, Insulin ; metabolism

OBJECTIVETo evaluate the clinical characteristics and risk factors of clonal evolution after immunosuppressive therapy (IST) in children with severe/very severe aplastic anemia (SAA/VSAA).

METHODSThe clinical data of 231 children with newly-diagnosed SAA/VSAA who received IST were retrospectively studied. The incidence and risk factors of clonal evolution after IST were analyzed.

RESULTSThe 5-year overall survival rate of the 231 patients was 82.7%. Except for 18 cases of early deaths, 213 patients were evaluated for IST efficacy. Among the 231 patients, cytogenetic abnormalities for at least two chromosome metaphase were detectable in 14 (7.4%) patients, and PNH clones were detectable in either peripheral red blood cells or neutrophils for 95 patients. Among the 213 patients evaluated for IST efficacy, 15 patients experienced clonal evolution after IST. Five patients had PNH and trisomy 8 which were defined as favorable progressions, and ten patients experienced monosomy 7 and MDS/AML as unfavorable progressions. The 5-year accumulative incidence of favorable and unfavorable progression were (2.2±2.2)% and (4.8±3.3)%, respectively. Until the last follow-up, 100% (5/5) of patients with favorable progressions and 50% (5/10) of patients with unfavorable progressions survived. WBC>3.5×10/L, CD3T cell percentage>80%, dosage of antithymocyte globulin >3.0 mg/(kg·d) and no response to IST were related to unfavorable progressions by univariate analysis. Cox multivariate analysis revealed that an increased CD3T cell percentage (>80%) and no response to IST were independent risk factors for unfavorable progressions.

CONCLUSIONSThe children with SAA/VSAA who have an increased CD3T cell percentage at diagnosis or have no response to IST are in high risks of unfavorable progressions.

Adolescent ; Anemia, Aplastic ; drug therapy ; genetics ; immunology ; mortality ; Child ; Child, Preschool ; Chromosome Aberrations ; Clonal Evolution ; Female ; Humans ; Immunosuppressive Agents ; therapeutic use ; Infant ; Male ; Proportional Hazards Models ; Retrospective Studies

OBJECTIVESTo dynamically observe how glioma stem cells promote the tumor formation and angiogenesis, and to study the correlation between the distribution of glioma stem cells and microvessels within different growth stages of subcutaneous tumor.

METHODSStem cell medium culture and magnetic activated cell sorting were carried out to obtain CD133+ cells from C6 rat glioma cell line. Sprague Dawley (SD) rat ears model were established to observe glioma stem cells promoting blood vessel formation. Subcutaneous glioma model of C6 and immunohistochemical staining of hypoxia inducible factor-1α (HIF-1α) and CD133 were used to investigate the relationship between distribution of glioma stem cells and microvessels. Expressions of CD133 protein in each stage of the subcutaneous tumor were detected by Western blot.

RESULTSIsolation and identification of glioma stem cells deprived from C6 glioma cell line successfully, the establishment of ears model showed real-time dynamic observation of CD133+ cells involved in angiogenesis and tumor formation. SD rat model of subcutaneous glioma showed the initial of tumor formation, CD133+ cells scattered. With tumor growth, CD133+ cells began to tend to capillaries, in late distributed clusters in perivascular. Meanwhile as tumor growth, CD133 protein expression was gradually increased: the values of Western blot analysis of CD133 expression on 6, 9, 12, 15, 20 d were 0.208±0.004, 0.282±0.003, 0.360±0.004, 0.564±0.135, 0.756±0.007, the differences were significant between different groups (F=2601.681, P<0.01). At a high magnification, the CD133 scores with immunohistochemical staining on 6, 9, 12, 15 d were 0.8±0.4, 2.4±0.5, 4.0 ± 0.7, 6.0±0.7; HIF-1α scores were 0.8±0.4, 2.8±0.8, 5.0±0.7, 6.8±0.4. By Spearman rank correlation analysis found that the relationship between CD133 and HIF-1α expression was positively correlated (r=0.921, P<0.01).

CONCLUSIONSGlioma stem cells promote angiogenesis more than non-stem cells; HIF-1α and its downstream gene product might mediate the distribution of glioma stem cells around the perivascular.

Animals ; Cell Line, Tumor ; Glioma ; blood supply ; metabolism ; pathology ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Microvessels ; pathology ; Neoplasm Transplantation ; Neoplastic Stem Cells ; pathology ; Neovascularization, Pathologic ; pathology ; Rats ; Rats, Sprague-Dawley