Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Breast Neoplasms ; metabolism ; pathology ; surgery ; Carcinoma, Lobular ; metabolism ; pathology ; surgery ; Female ; Histiocytoma, Malignant Fibrous ; metabolism ; pathology ; surgery ; Humans ; Immunohistochemistry ; Middle Aged ; Receptors, Progesterone ; metabolism ; Retroperitoneal Neoplasms ; metabolism ; pathology ; surgery

OBJECTIVETo study the correlation between arachidonic acid (AA) and acute red blood cells damage in rats, and to build a model with hidden blood loss in vivo, and to explore the pathological mechenism of hidden blood loss.

METHODSA total of 50 male adult Sprague-Dawley rats weighing (200 ± 20) g were randomly divided into five groups (n = 10): control group and four experimental groups. The rats in the experimental groups were given 0.5 ml different concentrations of AA dilu- ents, 5, 10, 20, 40 mmol/L respectively. The blood samples were collected from orbital venous at the beginning and 24, 48, 72 hours after administration. Then the changes of hemoglobin (Hb) ,red blood cell count (RBC), glutathione peroxidase (GSH- PX) activity, total superoxide dismutase (T-SOD) activity and hydrogen peroxide (H202) in the blood samples were tested.

RESULTSSignificant hidden blood loss occurred when the concentration was 10 mmol/L in the experimental group, with the RBC and Hb sharply reduced in blood samples. The Hb and RBC were reduced in all the experimental groups and control group at 24 hours after administration, while in the experimental groups they changed more obviously. The GSH-PX activity, T-SOD activity and H₂O₂were also significantly reduced in all groups, and the changes showed significant differences. The Hb and RBC were relatively stable in the control group and the experimental groups at 48 hours after administration; while GSH-PX activity, T-SOD activity and H₂O₂were all significantly decreased, and the changes in the experimental groups were more notable.

CONCLUSIONElevated levels of AA in the blood causes oxidative stress in the red blood cells, leading to the damage of red blood cells and hemoglobin, which is responsible for hidden blood loss.

Animals ; Arachidonic Acid ; toxicity ; Erythrocytes ; drug effects ; metabolism ; Glutathione Peroxidase ; blood ; Hemoglobins ; analysis ; Male ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; blood

Objective: To investigate the prevalence and influencing factors of adolescent depression symptoms in Zhejiang, so as to provide reference for improving their mental health. Methods: The middle school and university students in 11 cities of Zhejiang Province were selected by stratified cluster random sampling method. The depression symptoms of the adolescents were assessed by Center for Epidemiological Survey-Depression Scale ( CES-D ) and the influencing factors were analyzed by multivariate logistic regression model. Results: A total of 25 855 students were investigated, and 25 614 ( 99.07% ) valid questionnaires were collected. The detection rate of depressive symptoms was 26.86%(6 879 cases). The detection rate of depressive symptoms in girls was 29.75%, which was higher than 24.12% in boys ( P<0.05). The detection rate of depressive symptoms in high school students was 31.74%, the highest compared with other grades. The multivariate regression analysis showed that female students ( OR=1.690, 95%CI: 1.592-1.794 ), resident students ( OR=1.071, 95%CI: 1.010-1.137 ) , internet addiction ( OR=2.948, 95%CI: 2.527-3.439 ) , attempt smoking ( OR=1.516, 95%CI: 1.359-1.690 ), drinking ( OR=1.624, 95%CI: 1.525-1.729 ), bullied in the past 30 days ( OR=3.143, 95%CI: 2.938-3.363 and having serious injuries within a year ( OR=1.369, 95%CI: 1.263-1.543 ) were associated with adolescents who had depressive symptoms. Conclusions The detection rate of depressive symptoms is relative 26.86% among adolescents of Zhejiang Province. The students who are female, live on campus, have internet addiction, have been bullied or seriously injured, smoke and drink are more likely to have depressive symptoms.

Abstract: Objective - - To establish a pre column derivatization high performance liquid chromatography method for detecting Methods dimethyl sulfate (DMS) in workplace air. DMS in workplace air was collected with mercaptopyridine impregnated （ silicone tube. The derivative of DMS and mercaptopyridine was eluted by mobile phase phase A: water, phase B: acetonitrile, ∶ the volume ratio was 40 60) , and separated with a C18 column, then detected with diode array detector and quantitated by a Results - standard curve. The linear range of DMS was 0.17 40.00 mg/L, with the correlation coefficient of 0.999 95. The detection limit and the lower limit of quantitation were 0.05 and 0.17 mg/L respectively. The minimum detection concentration and minimum quantitation concentration were 0.02 and 0.04 mg/m³, respectively (air sample volume of 4.5 L, 1.0 mL sample - - - solution). The average desorption efficiency was 98.40% 102.00%. The within run and between run relative standard deviations - - were 0.61% 3.92% and 1.71% 6.00%, respectively. The samples could be stored at room temperature for at least 14 days. Conclusion This method can be used to detect DMS in workplace air.

Abstract: Nitrobenzene compounds (NBCs) are widely used in the world. It has 40 isomers such as nitrobenzene, dinitrobenzene and nitrotoluene, that are highly toxic and difficult to degrade and can cause harm to human health in different degrees. At pres⁃ ent, there is no unified standard method and occupational exposure limit for the detection of NBCs in the air. In terms of sampling medium, solid adsorption tube is mostly used for trapping vapor state NBCs, and filter membrane and solid adsorption tube are mostly used in series for sampling coexist NBCs in vapor state and aerosol state. In the detection methods, gas chromatography and liquid chromatography are common, and ultraviolet spectrophotometry, Raman spectroscopy, ion migration spectrometry and some other rapid response methods and technologies are also used in the detection of NBCs. In the detection of NBCs by gas chro⁃ matography, capillary column separation is commonly used, and the main detectors are flame ionization detector, electron capture detector and mass spectrometry detector. It is of practical significance to establish a method with high sensitivity, strong practica⁃ bility, convenient operation, and can simultaneously collect and detect a variety of NBCs in different states.

Objective To investigate the effect of chronic intermittent hypoxia(CIH)on the cardiovascular system in anesthetized rats. Methods A totally of 72 male SD rats were randomly divided into three groups(n=24),namely the ormoxia control group(control group), the normoxia anesthesia group(model group)and the chronic intermittent hypoxia group(CIH group).Rats of the control group breathe nor-mally.The model group was given intraperitoneal injection of 10%hydration with 0.3 mL/kg,and the CIH group was given chronic intermit-tent hypoxic stimulation with 8 h/d in addtion to the model group.The difference of ultrasonic echocardiography data,blood pressure,endothe-lin type-1,and endothelial nitric oxide synthase(eNOS)in rats of the three groups were compared.After 28 days,these rats were sacrificed to observe the changes of myocardial cell structure.Results In the control group,the myocardial morphology was normal,the cells arranged e-venly,and there was no swelling and inflammation.In the model group,the myocardial cells were evenly arranged without hypertrophy and in-flammatory changes.In the CIH group,the myocardial cells in the hypoxic group were not evenly arranged,and hypertrophy,swelling,deform-ation,hyperchromia,and obvious inflammatory changes of the myocardial tissue were observed.In the control group,myocardial cell nucleus and cytoplasm were uniformly arranged,and there was no obvious changes in the model group.On the contrary,myocardial cell morphology changed obviously in the CIH group,with the cell morphology and size of the inhomogeneity increased, and the color of the apoptosis cells changes from light to dark.The tail artery systolic pressure of rats in CIH group was significantly higher than that of the control group and the model group,and the LVEF of CIH group was significantly lower than that of the other two groups(P<0.05).Ultrasound detection value sug-gests that the LVID and left ventricular of rats in the CIH group were slightly larger,and the diastolic function was normal.The LVDs of the model group and the CIH group were both higher than that of the control group with statistically significant difference(P<0.05).The RBC, HCT,dp/dtmax,and -dp/dtmax in the CIH group were significantly higher than those of the control group and the model group(P<0.05). Serum levels of endothelin-1 in CIH group were significantly higher than that of control group and model group,while the summation of serum NO2-/NO3-and eNOS in CIH group were significantly lower than those of the control group and the model group(P<0.05).Conclusion Chronic intermittent hypoxia can cause cardiac dysfunction in anesthetized rats,which may lead to the imbalance of serum endothelin-1 and NO levels,leading to endothelial dysfunction and myocardial injury.

Objective - - To prepare the GDH 5 air sampling tube for simultaneous collection of eight kinds of chloro nitrobenzene （ ） ， compounds CNBs in the air of workplace and establish a matching determination method using gas chromatography. Methods - - ， Eight kinds of CNBs in vapor and aerosol state were collected by self developed GDH 5 air sampling tube desorbed ， ， ， by toluene separated by polysiloxane gas chromatography column detected by microcell electron capture detector and Results - （ - quantified by external standard method. It was determined that the air sampling tube was assembled by XAD 2 ion ） - ， exchange resin and glass fiber filter membrane. The linear range of CNBs was 0.80 240.00 mg/L and the linear correlation - - coefficients were greater than 0.999 9. The detection limit was 7.87 13.03 μg/L. The minimum detectable concentration was 0.60 3， - 3（ ） 1.33 μg/m and the minimum quantitative concentration was 2.00 4.22 μg/m sample 45.00 L . The average desorption - - （RSD） - ， - RSD efficiency was 101.2% 110.0%. The within run relative standard deviation was 0.8% 4.1% and the between run - Conclusion - was 0.3% 5.8%. The samples could be stored for more than 30 days at room temperature. GDH 5 air sampling tube and its associated determination method can be used for the collection and determination of eight kinds of CNBs in workplace air.

OBJECTIVECardiotonic steroids (CTS) can bind to Na+, K+ -ATPase to activate complex intracellular signaling cascades regulating the proliferation and apoptosis of cells. The aim of this study was to investigate the effects of ouabain at different concentrations on growth regulation in various kinds of leukemia cell lines and explore the pathogenesis of leukemia, the functions of Na+, K+ -ATPase as a signal transduction conductor and its effects on cell growth.

METHODSUsing the MTT assay, the survival rates of leukemia cell lines were observed 6, 12 and 24 hrs after treatment with 1 or 10 nmol/L ouabain. The expression of Na+, K+ -ATPase alpha1 subunit of leukemia cells was detected by Western blot.

RESULTSThe MTT results showed that ouabain at 1 nmol/L or 10 nmol/L induced proliferation of lymphocytic leukemia B95 and Jhhan cell lines, as well as megakaryocytic leukemia M07e and Meg01 cell lines. Ouabain at 1 nmol/L or 10 nmol/L increased the expression of Na+, K+ -ATPase alpha1 subunit. There were significant differences in the proliferation and the expression of Na+, K+ -ATPase alpha1 subunit of the leukemia cell lines between the ouabain treatment and the blank control groups 24 hrs after ouabain treatment (P<0.05). The proliferation effect of leukemia cell lines was in a direct proportion with the ouabain concentration and incubation time.

CONCLUSIONSNa+, K+ -ATPase plays an important role in signal transductions. Through binding to ouabain, Na+, K+ -ATPase may regulate proliferation of leukemia cell lines of different origins. Ouabain at 1 nmol/L or 10 nmol/L may induce proliferation of lymphocytic leukemia cell lines (B95, Jhhan) and megakaryocytic leukemia cell lines (M07e, Meg01), and the proliferation effect was in a direct proportion with the concentration and incubation time of ouabain.

Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Humans ; Leukemia ; pathology ; Ouabain ; pharmacology ; Signal Transduction ; Sodium-Potassium-Exchanging ATPase ; analysis ; genetics ; physiology

OBJECTIVETo observe the protective effects of Tongxinluo (TXL) on apoptosis of rat cardiac microvascular endothelial cells (RCMECs) resulting from homocysteine (Hcy) induced endoplasmic reticulum stress (ERS), and to determine the signaling pathway behind its protection.

METHODSPrimary cultured RCMECs were isolated from neonatal rats using tissue explant method. The morphology of RCMECs was observed using inverted microscope, identified and differentiated by CD31 immunofluorescence method. Selected were well growing 2nd-4th generations of RCMECs. The optimal action time was determined by detecting the expression of glucose regulated protein 78 (GRP78) using immunofluorescence method. In the next experiment RCMECs were divided into 5 groups, i.e., the blank control group, the Hcy induced group (Hcy 10 mmol/L, 10 h), the Hcy + TXL group (Hcy 10 mmol/L + TXL 400 µg/mL), the Hcy +LY294002 group (Hcy 10 mmol/L + LY294002 5 µmol/L, LY294002 as the inhibitor of PI3K), the Hcy + LY294002 + TXL group (Hcy 10 mmol/L + LY294002 5 µmol/L + TXL 400 µg/mL). The apoptosis rate of RCMECs was detected by flow cytometry. mRNA and protein expressions of GRP78, C/ EBP homologous protein (CHOP), and cysteinyl aspartate specific proteinase-12 (caspase12) were detected by real-time reverse transcription PCR (RT-PCR) and Western blot respectively. Expression levels of phosphorylation of phosphatidylinositol 3-kinase (P-PI3K), total phosphatidylinositol 3-kinase (T- P13K) , phosphorylation of kinase B (P-Akt) , and total kinase B (T-Akt) were detected by Western blot.

RESULTSTen hours Hcy action time was determined. Compared with the blank control group, the apoptosis rate was increased (22.77%), mRNA and protein expressions of GRP78, CHOP, and Caspase-12 were increased, protein expressions of P-PI3K and P-Akt,ratios of P-PI3K/T-PI3K and P-Akt/T-Akt were decreased in the Hcy induced group (P < 0.05, P < 0.01). Compared with the Hcy induced group, the apoptosis rate was decreased (10.17%), mRNA and protein expressions of GRP78, CHOP, and Caspase-12 were decreased, and expression levels of P-PI3K, P-Akt, P-PI3K/T-PI3K, and P-Akt/T-Akt were increased in the Hcy + TXL group (P < 0.05, P < 0.01). Compared with the Hcy + TXL group, the apoptosis rate was increased (17.9%), mRNA and protein expressions of GRP78, CHOP, and Caspase-12 were increased, expression levels of P-PI3K and P-Akt, ratios of P-PI3K/T-PI3K and P-Akt/T-Akt were decreased in the Hcy + TXL + LY294002 group (P < 0.05, P < 0.01).

CONCLUSIONTXL could inhibit the apoptosis of RCMECs resulting from Hcy-induced ERS and its mechanism might be associated with activating PI3K/Akt signaling pathway.

Animals ; Apoptosis ; drug effects ; Caspase 12 ; metabolism ; Cells, Cultured ; Chromones ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Endoplasmic Reticulum Stress ; Endothelial Cells ; drug effects ; Morpholines ; pharmacology ; Myocardium ; cytology ; Phosphatidylinositol 3-Kinases ; metabolism ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; metabolism ; Rats ; Signal Transduction ; Transcription Factor CHOP ; metabolism

OBJECTIVETo isolate single chain antibody fragments (scFv) against cell surface molecules by pathfinder selection from an anti-KG1a cell scFv phage library.

METHODSThe anti-KG1a scFv library was enriched by KGla cell panning for three rounds, or unenriched, then processed for pathfinder selection respectively using anti-CD34 monoclonal antibody as pathfinder molecule. ScFv phage clones were randomly picked and identified by binding KG1a cells using immunofluorescein and flow cytometry. The KG1a+ clones were further identified by KG1a, HL60, U937, and CEM cell lines and ELISA. Their antigenic molecules on cell surface were digested by chymopapain and analyzed by flow cytometry. DNAs from ten positive clones were sequenced. The scFv clones with different primary structure were used to analyze the molecular weight of their antigens by Western blot.

RESULTSOne hundred and two KG1a+ scFv phage clones were isolated from 144 enriched and 96 unenriched scFv phage library respectively, among which 47 bound KG1a, HL60, U937, and CEM cells, 55 bound KG1a cells exclusively. None of 28 KG1a+, HL60-, U937-, and CEM- scFv clones bound to the CD34 antigen, as confirmed by ELISA, although most of their antigens were sensitive to chymopapain digestion. DNA sequences from ten positive clones showed that they were from four different clones. They bound antigens with different molecular weight.

CONCLUSIONSOne hundred and two scFv phage clones specific for hematopoietic stem and progenitor cells have been isolated from an anti-KG1a cell scFv phage library. The pathfinder selection has showed advantages to improve the screening efficacy of scFv phage clones against antigens, which present at very low densities on the cell surface.

Antibodies, Anti-Idiotypic ; immunology ; Antibodies, Monoclonal ; genetics ; Antibody Specificity ; Bacteriophages ; genetics ; Cloning, Molecular ; Hematopoietic Stem Cells ; immunology ; Humans ; Immunoglobulin Fc Fragments ; biosynthesis ; genetics ; immunology ; Immunoglobulin Fragments ; genetics ; immunology ; Immunoglobulin Variable Region ; genetics ; immunology ; Peptide Library ; Single-Chain Antibodies