Objective To explore the source of endothelial cells in tumor vessels by A 549 tumor model with GFP nude mouse.Methods To establish the A 549 lung cancer models with GFP nude mice, expression of CD 31 was determined by immunofluorescence to label tumor vessels; to observe and take a picture of the tumor frozen section by confocal microscopy and invert microscope;expression of GFP in tumor vessels was determined by immunohistochemistry. Result The results of immunofluorescence showed:Tumor interstitial vascular endothelial cells or endothelial cells clusters and micro-vascular lumen size and shape are clearly visible by immunofluorescence, and part of vessels with no obvious lumen or irregular lumen.We can see green fluorescent in tumor cells of tumor tissue and endothelial cells which form of tumor vessels.The results of immunohistochemistry showed: expression of GFP was determined in cytoplasm of tumor stromal cells and endothelial cells in tumor vessels.Conclusion The endothelial cells which formed tumor neovessels that derived from GFP nude mice partly and the other part derived from tumor cells.

Objective To explore the expression of silencer of death domains(SODD) and its clinical significance and relationship with phospho-NF-?B-p65 proteins in bone marrow cells of acute lymphoblastic leukemia(ALL)in children,and the expression of SODD and phospho-NF-?B-p65 in Jurkat cells treated with chemotherapeutic drugs in order to find a new chemotherapeutic target.Methods The expressions of SODD and phospho-NF-?B-p65 proteins in bone marrow cells were detected by immunohistochemistry in 25 children with ALL.The apoptosis incidence was measured by Annexin-V-Fluorescence/PI double-labeling flow cytometry and the expression of SODD and phospho-NF-?B-p65 proteins were determined by Western blotting in Jurkat cells.Results It was found that the expression of SODD and active p65 expression in ALL were significantly higher than those in healthy control group.The expression of SODD and phospho-NF-?B-p65 proteins in the high-risk(HR) group was significantly higher than those in standard-risk(SR) group(Pa

Activation of acid-sensing ion channels (ASICs) plays an important role in neuroinflammation.Macrophage recruitment to the sites of inflammation is an essential step in host defense.ASIC 1 and ASIC3 have been reported to mediate the endocytosis and maturation of bone marrow derived macrophages.However,the expression and inflammation-related functions of ASICs in RAW 264.7 cells,another common macrophage,are still elusive.In the present study,we first demonstrated the presence of ASIC 1,ASIC2a and ASIC3 in RAW 264.7 macrophage cell line by using reverse transcriptase polymerase chain reaction (RT-PCR),Western blotting and immunofluorescence experiments.The non-specific ASICs inhibitor amiloride and specific homomeric ASIC 1 a blocker PcTx 1 reduced the production of iNOS and COX-2 by LPS-induced activating RAW 264.7 cells.Furthermore,not only amiloride but also PcTx 1 inhibited the migration and LPS-induced apoptosis of RAW 264.7 cells.Taken together,our findings suggest that ASICs promote the inflammatory response and apoptosis of RAW 264.7 cells,and ASICs may serve as a potential novel target for immunological disease therapy.

Objective To study the effect of lipopolysaccharide(LPS)on the endo-pulmonary natrium channel(ENaC)expression in the lung of rats with acute lung injured.Method Sixteen rats were randomly divided into normal control group and LPS-group.Rats of normal control group and LPS-group were killed at 6 hours after intravenous injection of normal saline(8 ml/kg)or LPS(8 mg/kg).The extent of lung injury was assessed by arterial blood gas analysis and histological examination.At the same time,?-ENaC protein and???- ENaC mRNA expression in the lung tissue were analyzed by immunohistochemistry and RT-PCR.Results PaO_2 in LPS-group was noticeably lower than in normal control group(P

This study was aimed to investigate the changes of silencer of death domains (SODD), survivin, caspase 3, caspase 8 and caspase 9 in the apoptotic process of human leukemia cells induced by chemotherapeutic drugs in order to explore the molecular mechanism of apoptotic modulatory genes and to search for the new target of chemotherapeutic drugs. After Jurkat cells were induced by chemotherapeutic drugs, the translocated phosphatidylserine was labeled with annexin V/PI, and the apoptosis incidence was measured by FCM; The expression changes of SODD, caspase 3, caspase 8 and caspase 9 were determined by Western blot; the changes of survivin mRNA and protein were determined by RT-PCR and immunohistochemistry SABC method respectively. The results indicated that high expressions of SODD and survivin could inhibit apoptotic signaling pathway; VCR down-regulated the function of SODD protein and effectively induced the apoptosis of Jurkat cells in a time-dependent manner and activates caspase 3 through the death receptor-mediated activation of caspase 8, in which caspase 9 and survivin were not degraded. It is concluded that SODD participates in the apoptotic process induced by VCR which induces the Jurkat cell apoptosis by downregulating expression of SODD protein and priming death receptor pathway. In the apoptotic process, the mitochondrion apoptotic pathway is not trigged.
Adaptor Proteins, Signal Transducing ; metabolism ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Humans ; Inhibitor of Apoptosis Proteins ; Jurkat Cells ; Microtubule-Associated Proteins ; metabolism ; Vincristine ; pharmacology

Objective To study whether the green fluorescent protein ( GFP) gene can be successfully expressed in green fluorescent nude mice and the tissue distribution characteristics.Methods Small animal imaging system and RT-PCR assay were used to detect the GFP tissue distribution and fluorescence expression level.Results The GFP can be expressed in multiple tissues in green fluorescent nude mice.A higher expression was observed in the pancreas, heart, brain, and skin.Conclusion Exogenous GFP can be stably expressed and inherited in green fluorescent nude mice, with the highest expression in the pancreas.

OBJECTIVESince 2011 EB-APS conference, we hypotheses that phase switching of inspiration-expiration is dominantly initiated by oscillatory information PaO2, PaCO2 and [H+] via fast peripheral chemical receptors. However, the evidence of the waveform of ABG is lack.

METHODSSix surgery patients with normal heart function and negative Allen test, had been placed the arterial catheterization directly connected to 3 x 1 000 mm pre-heparin plastic pipe for continuous collecting arterial blood. We counted the number of heart beat for the blood collecting time, and separated the blood pipe into the heart beat numbers' short pieces using haemostatic forceps, then put pipe into iced water at once fir analyzing PaO2, PaCO2, pH and SaO2 as soon as possible. We selected two breaths cycles of waveform from each patient for data calculations of magnitudes and time interval.

RESULTSThe heart beat numbers for filling blood into pipe were 16 ± 2, and all covered more than 2 breathing cycles. Each breathing cycle is cover 5 ± 0.6 heart beat. There were significant changes of PaO2, PaCO2, [H+] a and SaO2 (i.e. the highest high values compare to the next lowest values, P < 0.05). The time interval of changing PaO2, PaCO2, [H+]a and SaO2 magnitudes were 11.28 ± 1.13 mmHg, 1.77 ± 0.89 mmHg, 1.14 ± 0.35 nmol/L and 0.52% ± 0.44% respectively.

CONCLUSIONThis simple continuous beat-by-beat arterial blood sampling and ABG analyzing method is new and practicable. We obtain a clear evidence of periodic parameters ABG waveform, which following breathing cycle.

Arteries ; physiology ; Blood Gas Analysis ; Heart Rate ; Humans ; Monitoring, Physiologic ; methods ; Respiration

OBJECTIVETo analyze the cervical cancer mortality and distribution profiles by areas and population in 2004 - 2005 and the changes in the last 30 years.

METHODSThe demographic information and cervical cancer death data came from the Third National Retrospective Sampling Survey of Death Causes which including 158 sampling areas (including 61 cities and 97 villages; 52 in the east region, 49 in the middle region and 57 in the west region) covered 31 province-level regions in the interior of China by multiple stage stratified clustering sampling. The total number of the samples were 142 660 482 person years (72 970 241 person years in male and 69 690 241 person years in female). The crude and age-specific death rate was calculated. The standardized death rate was calculated by using 5 years interval from the census data of 2000. In addition, the mortality data was compared with the two previous national death surveys in 1970's and 1990's, respectively.

RESULTSIn the sampling region of the third retrospective death causes survey from 2004 to 2005, the crude death rate of cervical cancer was 2.86/100 000 (1995/69 690 241), which accounting for 2.86% (1995/69 667) and ranking 9th of all cancer death causes in female population. The crude death rates of cervical cancer were 2.88/100 000 (1326/46 091 419) and 2.83/100 000 (669/23 598 822) for rural areas and urban areas, respectively, the Chinese population adjusted death rates of rural and urban areas were 2.01/100 000 and 1.67/100 000, respectively. The standardized death rates of cervical cancer in 1973 - 1975 and 1990 - 1992 were 11.10/100 000 and 3.25/100 000, respectively, which accounting for 17.91% and 4.86%, respectively; in 2004 - 2005, the crude death rates increased by age, rose remarkably from the age of 40 and arrived the peak of 20.83/100 000 (82/393 624) at the age of 85. The mortality among 35 - 44 year-old group was higher than those in 1990's. The crude death rates of the middle, west and east areas were 3.41/100 000 (827/24 225 738), 3.25/100 000 (636/19 563 647) and 2.05/100 000 (532/25 900 856), respectively, the Chinese population adjusted death rates of the middle and west areas were about twice that of the east (2.35/100 000 in the middle, 2.38/100 000 in the west, and 1.19/100 000 in the east).

CONCLUSIONThe mortality of cervical cancer was higher in rural areas than that in urban, as well as higher in the middle and west areas than that in east area in the interior of China in 2004 - 2005. Younger trend of cervical cancer death was observed; the mortality of cervical cancer and constituent ratio in the female tumor death continued to decline compared to those in 1973 - 1975 and 1990 - 1992.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Cause of Death ; trends ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Health Surveys ; Humans ; Infant ; Infant, Newborn ; Middle Aged ; Neoplasms ; Uterine Cervical Neoplasms ; epidemiology ; mortality ; Young Adult

OBJECTIVETo investigate the effects of Tounongsan () extract (TNSE) on proliferation and apoptosis of the human lymphoma cell line Raji and its possible mechanism of action.

METHODSThe viability of TNSE-treated Raji cells was measured by a 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Cell apoptosis was determined by flow cytometry. The molecular mechanisms of TNSE-mediated apoptosis were further investigated by reverse transcription-polymerase chain reaction (RT-PCR) analysis of the mRNA expression of nuclear factor κB (NF-κB), Bcl-xL, Bcl-2-associated death promoter (Bad), caspase-9 and caspase-3. Western blotting was used to detect the protein expressions of NF-κB, Bad, cleaved caspase-9 and cleaved caspase-3.

RESULTSTNSE inhibited Raji cell proliferation in dose- and time-dependent manners. After 48-h treatment with various concentrations of TNSE (125, 250 and 500 μg/mL), the apoptosis rates of Raji cell were 12.23%±1.98% (P<0.05), 20.97%±3.96% (P<0.01) and 30.4%±4.87% (P<0.01), respectively, compared with those of the control (6.02%±1.01%). RT-PCR demonstrated that NF-κB mRNA expression was significantly downregulated in Raji cells treated with 250 μg/mL TNSE for 48 h (P<0.05), while Bad, caspase-9 and caspase-3 mRNA levels were upregulated (P<0.05). Moreover, TNSE treatment resulted in downregulation of NF-κB protein expression and strikingly upregulated protein expressions of Bad, cleaved caspase-9, cleaved caspase-3 in a dose-dependent manner, as determined by Western blot.

CONCLUSIONTNSE exhibits significant anti-proliferative and apoptotic effects in Raji cells, which may be involved in regulation of NF-κB and Bad, and activation of caspase-9 and caspase-3.

Apoptosis ; drug effects ; Caspase 3 ; genetics ; metabolism ; Caspase 9 ; genetics ; metabolism ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; NF-kappa B ; genetics ; metabolism ; Plant Extracts ; pharmacology ; Tumor Cells, Cultured ; bcl-Associated Death Protein ; metabolism

OBJECTIVETo study the effect of intrahepatic cholestasis of pregnancy (ICP) on the functions of the hypothalamic-pituitary-adrenocortical (HPA) axis and adrenal cortex in normal neonates.

METHODSDemographic characteristics, prenatal anxiety and depression, and perceived stress during delivery were investigated in 32 ICP women and 32 controls. The cord blood levels of cortisal, adrenocorticotropic hormone (ACTH), and dehydroepiandrosterone sulfate (DHEAS) were measured by the radioimmunity technique in normal neonates immediately after birth.

RESULTSThe scores of prenatal anxiety and depression in ICP women were significantly higher than those in controls (p<0.05 and p<0.01, respectively). There were no significant differences in the perceived stress during delivery between the two groups. The cord blood levels of cortisol and ACTH in neonates from ICP women were significantly lower (p<0.01), while the DHEAS level was significantly higher (p<0.01) than in neonates from controls. The DHEAS/ACTH ratio was significantly higher (p<0.01), while the cortisol/DHEAS ratio was significantly lower in the ICP group (p<0.01) than in the control group. The glycocholic acid level in ICP women was positively correlated with the DHEAS level in neonatal cord blood (r=0.47, p<0.01).

CONCLUSIONSThere may be a dissociation between cortisol and DHEAS in neonates with normal birth outcome from ICP women. ICP may result in a decreased responsiveness of HPA axis and an increased secretion of DHEAS by adrenal cortex in these neonates. This suggests that there might be dysfunction of the fetal zones of the adrenal cortex.

Adrenal Cortex ; physiopathology ; Adrenocorticotropic Hormone ; blood ; Adult ; Cholestasis, Intrahepatic ; physiopathology ; Dehydroepiandrosterone Sulfate ; blood ; Female ; Humans ; Hydrocortisone ; blood ; Hypothalamo-Hypophyseal System ; physiopathology ; Infant, Newborn ; Pituitary-Adrenal System ; physiopathology ; Pregnancy ; Pregnancy Complications ; physiopathology