Objective To investigate effects of Kanglaite injection on proliferation, cycle and apoptosis of human breast cancer MCF-7 cells;To discuss its relevant mechanism. Methods Logarithmic growth phase cells were divided into control group and Kanglaite-treatment group (10, 20, 40μL/mL). Cells were cultured in RPMI-1640 for 24 h before drug treatment. The inhibition rate of Kanglaite injection on proliferation of human breast cancer MCF-7 cells was detected by MTT assay. Apoptosis and cell cycle of MCF-7 cells were detected by flow cytometry. Changes in cell nucleus were determined by Hochest staining assay. Protein expressions of Bcl-2 and Bax were detected by ELISA and Western blot. Results Kanglaite injection for 12 h, 24 h or 48 h resulted in a significant inhibition of MCF-7 cells proliferation (P<0.05, P<0.01);Compared with the control group, Kanglaite injection-treated cells showed increased percentage in G2/M and G0/G1 phases (P<0.001, P<0.01), but showed decreased percentage in S phase (P<0.01), and apoptosis rate increased (P<0.05, P<0.001). Kanglaite injection significantly decreased protein expression of Bcl-2, and enhanced protein expression of Bax of MCF-7 cells (P<0.01, P<0.001). Conclusion Kanglaite injection can inhibit the proliferation of human breast cancer MCF-7 cells, decrease cell cycle and induce apoptosis, the mechanism is related with decreasing protein expression of Bcl-2 and enhance the protein expression of Bax.

Objective To investigate the effects of S100B on the expressions of dopamine receptors and the synthesis,metabolism of neurotransmitters dopamine,5-hydroxytryptamine which are related to the abnormal motor coordination of Parkinson's disease (PD).Methods The hS100B transgenic mice were established.The mice were divided into S100B transgenic group (TG,n =14),S100B knockout group (KG,n =14) and the non-transgenic control group (CG,n =14).The motor coordination ability of mice was measured by the Rota-rod test.The expressions of dopamine D1 receptor (D1DR),dopamine D2 receptor (D2DR),tyrosine hydroxylase (TH) and phosphorylated TH at Ser19,Ser31,Ser40 in brain tissue were detected by reverse transcription polymerase chain reaction and Western blotting.The levels of Tyr,levodopa,dopamine,homovanillic acid,Trp,5-hydroxytryptamine and 5-hydroxyindoleacetic acid in mesencephalon were measured by high performance liquid chromatography with fluorescence detection.Results Compared with CG,the motor coordination ability of mice (s) showed progressive decline in TG (3 months:4.60±0.30vs4.25±0.21,q =5.194;6 months:4.52±0.31 vs4.07±0.22,q =6.139;9 months:4.43 ± 0.25 vs 3.60 ± 0.18,q =13.484;all P ＜ 0.05),the expressions of D2DR mRNA and protein decreased (1.34 ± 0.13 vs 0.48 ± 0.07,q =21.578;1.05 ± 0.15 vs 0.69 ± 0.10,q =8.063,both P＜0.05) and phosphorylated TH at Serl9 and Ser40 increased (0.95 ±0.10 vs 1.14-0.13,q =4.972;0.94 ± 0.12 vs 1.17 ± 0.14,q=5.382,both P＜ 0.05),the levels of levodopa,dopamine and homovanillic acid were elevated (87.04 ± 11.77 vs 115.28 ± 16.80,q =4.764;56.66 ± 9.87 vs 72.96 ± 11.02,q=3.923;26.58 ± 8.11 vs 38.65 ± 6.67,q=3.981,all P＜ 0.05),the leve1 of 5-hydroxytryptamine was reduced (925.50 ± 74.26 vs 637.87 ± 56.76,q =11.084,P ＜ 0.05),the ratios of homovanillic acid/dopamine and 5-hydroxyindoleacetic acid/5-hydroxytryptamine increased (0.45 ± 0.05 vs 0.54±0.08,q =3.325;0.94±0.07 vs 1.42±0.12,q =12.367,both P＜0.05) in the brain of TG at the age of 9 months old.There was no significant difference of detection indexes between KG and CG.Conclusions S100B plays an important role in the development of PD and the brain-specific S100B transgenic mice can be used to investigate the function of S100B gene on the development of PD.

Objective To investigate the expression and distribution of the downstream substrate of extracellular regulated protein kinase(ERK1/2) pathway, ternary complex factor phospho-Elk-1, in rat brains with chronic fluorosis, and reveal the mechanism of the impaired learning and memory ability caused by chronic fluorosis. Methods Seventy-two SD rats, weighing 100 - 120 g, were randomly divided into 3 groups, 24 in each group (half male and half female). The rats in control group were fed with tap water (fluoride < 0.5 mg/L); low- and high-dose fluoride groups were fed with tap water with different concentrations of NaF(5.0,50.0 mg/L F-, respectively). After 6 months, body weight was weighed, dental fluorosis was determined by observation and urinary fluoride and bone fluoride were detected by fluorine ion-selective electrode; the learning ability of rats was measured by navigation test of Morris water maze, and memory ability by spatial probe test in Morris water maze; the expression and distribution of phospho-Elk-1 in different brain regions were detected by immunohistochemistry method. Results In low- and high-fluoride groups, the body weight of rat[(449.2 ± 77.1), (312.8 ± 89.7)g] was significantly decreased than that of control [(635.5 ± 76.2 )g, all P< 0.05], the varying degrees of dental fluorosis were observed(x2 = 7.83, P<0.05), urinary fluoride[(2.56 ±0.91),(5.73 ±3.14)mg/L] and bone fluoride[(709.2 ± 37.4) ,(1306.3 ± 102.4) mg/kg] were significantly higher than those in controls[(0.92 ± 0.30)mg/L,(348.5 ± 89.2)mg/kg, all P< 0.05]. The escape latency of low- and high-fluoride groups[ (7.4 ± 4.1), (12.2 ± 5.7)s] was longer than that of control [(4.8 ± 2.7 )s, all P < 0.05] and the escape latency in high-fluoride group was significantly longer than that in other groups (all P < 0.05); in spatial probe test, the time of first crossing platform was longer in rats with fluorosis [(4.18 ± 1.10),(5.89 ± 0.56)s] as compared to control[(1.17 ± 0.75)s, all P< 0.05]. Expressions of phospho-Elk-1 in the hippocampus CA1(167.4 ± 8.3,163.2 ± 9.4), CA2(175.7 ± 5.0,183.3 ± 4.2), CA3(165.2 ± 11.6,162.9 ± 4.4), CA4(168.7± 6.9,169.5 ±5.3), fascia dentate (185.2 ±4.0,193.1 ±6.1) and caudate putamen( 181.4 ± 3.8, 179.8 ± 5.5) in low- and high-fluoride groups were higher than those of controls(142.4 ± 8.1,144.9 ± 8.4,143.6 ± 5.8, 116.8 ± 9.1,140.2 ± 7.8,163.1 ± 13.1, all P< 0.05). Conclusion Chronic fluorosis can cause increased expression of phospho-Elk-1 in the hippocampus and caudate putamen region of rat brains, which might be related to the mechanisms of decreased learning and memory ability of rats overexposed to fluoride.

To study the contribution of T cell subsets in the pathogenesis of Murine hepatitis virus Type3 (MHV-3) induced chronic viral hepatitis in C3H/Hej mice, ninety C3H/Hej mice were chosen to individually receive 10 plaque forming units (PFU) of MHV-3 intraperitoneally. The changes of virus titer and pathology in liver tissue were examined by standard plaque assay and by the hematoxylin/eosin (HE) staining method from 2 days post MHV-3 infection. The ratios of T cell subsets including CD3+CD4+CD8-, CD3+CD4-CD8+, CD3+CD4-CD8-, CD3+CD4+CD25+, CD3+CD4+CD25- and CD3+CD4-CD25+ T lymphocyte of total T lymphocytes in blood, spleen and liver were examined at 0, 2, 4, 6,8, 10, 12, 15, 20, 25, 30, 40 days post MHV-3 infection by flow cytosorting. We observed that the virus titer raised and showed persistent virus duplications and inflammatory changes in the livers of C3H/Hej mice from 2 days post MHV-3 infection. The double negative T cell (DN Treg cell) and CD4+CD25+ T cell ratios increased significantly from 2 days post MHV-3 infection in C3H/Hej mice, and CD3+CD4+CD8-, CD3+CD4-CD8+, CD3+CD4+CD25- and CD3+CD4-CD25+ T cell ratios decreased accordingly. In conclusion, the changes of virus titer and pathology in the livers of C3H/Hej mice post MHV-3 suggest their contribution to viral persistence. Further characterizations of DN Treg cells are that infection indicates that MHV-3 could induce the chronic inflammation in livers of C3H/Hej mice.The increase of the DN Treg cell and CD4+CD25+ T cell ratios in C3H/Hej mice post MHV-3 infection suggests that DN Treg cells and CD4+CD25+ T cells may both have important suppressive immunomodulation functions in the development of chronic viral hepatitis and have important roles in the virus persistent infection. Further characterizations of DNT cell and CD4+CD25+ T cell are under investigation.

Objective To evaluate the role of JNK signal pathway in brain injury after resuscitation in a rat model of asphyxia cardiac arrest.Methods Forty healthy male SD rats 'weighing 300-350 g were randomly divided into 4 groups ( n =10 each):sham operation group (group SH) ; cardiac arrest group (group CA) ; group SP600125-JNK inhibitor (group SP) and dimethyl sulfexide (DMSO) group.The rats were anesthetized with intraperitoneal pentobarbital 45 mg/kg,tracheostomized and mechanically ventilated.PETCO2 was maintained at 35-45 mm Hg.Femoral artery and vein were cannulated for BP monitoring and fluid infusion.Cardiac arrest was induced by clamping tracheal tube until ECG activity disappeared and MAP ＜ 10 mm Hg.Resuscitation was started at 3 min after cardiac arrest.MAP ＞ 60 mm Hg and HR ＞ 250 bpm were considered to be signs of successful resuscitation.SP600125 20 mg/kg and DMSO 0.2 ml were injected iv as soon as chest compression was started in groups SP and DMSO respectively.The animals were sacrificed at 5 h after successful resuscitation and their brains were removed for determination of wet/dry (W/D) weight ratio and microscopic examination of hippocampus.Neuronal apoptosis was detected by TUNEL.Results Cardiac arrest significantly increased W/D ratio and the number of apoptotic cells in group CA.SP600125 iv significantly attenuated the cardiac arrest-induced increase in W/D ratio and the number of apoptotic cells but DMSO did not.Conclusion JNK signal pathway is involved in the brain injury after resuscitation in a rat model of asphyxia cardiac arrest.

A sensitive determination method for sulfur mustard ( HD ) metabolites thiodiglycol ( TDG ) in rabbit urine was established and validated using isotope dilution negative ion chemical ionization ( NICI) gas chromatography-mass spectrometry ( GC-MS ) , in which deuterated thiodiglycol ( TDG-d8 ) was used as internal standard. Two solid-phase extraction ( SPE) steps were established and optimized in order to reduce the interfering backgrounds, one was used to extract thiodiglycol ( TDG ) from urine with self-assemblied Florisil SPE cartridges, another cleaning treatment of the by-products after pentafluorobenzoyl chloride (PFBZ) derivatization. The results showed that the limits of detection quantitation of this method were 0. 1 and 0. 3 μg/L, respectively. The exposure time-response relationship and exposure dose-response relationship of TDG in rabbit urine were studied after rabbit skin exposure to sulfur mustard (HD, 0. 02-0. 15 LD50). The TDG levels in the rabbit urine increased rapidly during the first day after application and then decreased over time for all dosage groups. A secondary release was also noted for the high-dose group, and the duration of high TDG excretion levels was correlated positively with the HD dosage levels. We thus concluded that abnormally high levels of TDG in urine could be used as a clear diagnostic indicator of HD exposure.

Objective To analyze the investigation results of an acute respiratory infection outbreak caused by adenovirus and provide scientific information for the prevention and control of congener public health emergencies .Methods A case‐control study was performed with grades and gender as matching factor ,all cases and selected controls were investigated with the same question‐naire .Results A totul of 47 cases were diagnosed in the outbreak ,no death ,the attack rate was 8 .88% ;the main clinical symptom was fever and 27 .7% of the cases became pneumonia .The case‐control study analysis demonstrated that with close contact to cases or not(χ2 =7 .96 ,P<0 .05) ,contact time (χ2 =7 .95 ,P<0 .05) ,hand washing habits (χ2 =25 .92 ,P<0 .05) and with or without the habit of cleaning snivel by hand directly (χ2 =22 .78 ,P<0 .05) were statistically different between cases and controls .Conclu‐sion long‐time contact to cases maybe the main risk factor for the adenovirus infection ,especially the contact manner were sharing the same desk or playing together .A good health habit of washing hands often and no cleaning snivel by hand directly were impor‐tant protective factors .Thus ,strengthening the training of health habit and awareness is the important preventive measure for re‐spiratory infectious diseases .

OBJECTIVETo investigate the effect of paraquat (PQ) on reactive oxygen species (ROS) and neutrophil apoptosis and its possible signal transduction pathways.

METHODSCultured neutrophils were treated with different concentrations of PQ for 6-24 h. The apoptosis rate of neutrophils and ROS content were determined by flow cytometry. The exoressions of nuclear factor kappa B (NF-κB) and Caspase 3 were detected by Western blot. These parameters were checked again after NF-κB and Caspase 3 antagonist were applied.

RESULTSPQ could boost ROS generation and depress neutrophil apoptosis significantly. At the same time PQ could enhance the expression of NF-κB and inhibit the expression of Caspase 3. These effects could be reversed by ROS inhibitor diphenyleneiodonium (DPI) and NF-κB inhibitor pyrrolidinedithiocarbamate (PDTC).

CONCLUSIONPQ is a potent inducer of ROS and can inhibit neutrophil apoptosis by activating NF-κB and surpressing Caspase 3 activity.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cells, Cultured ; NF-kappa B ; antagonists & inhibitors ; metabolism ; Neutrophils ; cytology ; drug effects ; Paraquat ; toxicity ; Pyrrolidines ; pharmacology ; Reactive Oxygen Species ; metabolism ; Signal Transduction ; Thiocarbamates ; pharmacology

Objective To evaluate the role of c-Jun N-terminal kinase (JNK) in lipopolysaccharide (LPS)-induced acute lung injury ( ALI) in rats.Methods Eighty male SD rats weighing 250-300 g were randomly divided into 4 groups ( n = 20 each) : control group (group C) ; ALI group; LPS + SP600125 (JNK inhibitor)group (group S) and LPS+ DMSO (the solvent) group (group DMSO) . ALI was induced by intravenous LPS 5mg/kg. In S and DMSO groups, SP600125 30 mg/kg and DMSO 0.2 ml were injected intravenously after LPS administration respectively. Ten animals were sacrificed by exsanguinafions at 4 h after LPS administration in each group. The broncho-alveolar lavage fluid (BALF) was colleted. The TNF-α and IL-1β concentrations in BALF were measured. The lungs were removed for microscopic examination and determination of W/D lung weight ratio. The other 10 animals in each group were observed for 48 h survival rate. Results Intravenous LPS significantly increased TNF-α and IL-1β concentrations in BALF and W/D lung weight ratio, decreased 48 h survival rate and induced histologic damage. Intravenous SP600125 30 mg/kg significantly attenuated the above-mentioned LPS-induced changes. Conclusion Activation of JNK is involved in the development of endotoxin-induced ALI in rats.

Objective To investigate the diagnostic value of the T-SPOT.TB test for diagnosing tuberculous meningitis(TBM) by meta-analysis.Methods A systematic retrieval from the databases of PubMed,EMBASE,etc.was performed.The literature on the T-SPOT.TB test for diagnosing TBM was collected.Two reviewers independently screened the literature,extracted the data and judged the quality.The meta analysis was conducted by the Meta-Disc 1.4 software.Results 8 articles were included,involving 425 patients including 232 cases of TBM.In the peripheral blood group,the combined sensitivity was 80%(95%CI:0.74-0.85),the combined specificity was 74%(95%CI:0.67-0.80),the area under the curve(AUC)of summary receiver operating characteristic (SROC)was 0.858 7;the diagnostic odds ratio(DOR)was 15.50.In the CSF group,the combined sensitivity was 76%(95%CI:0.70-0.82),the combined specificity was 83%(95%CI:0.77-0.88),AUC was 0.892 7;DOR was 22.62.Conclusion Adopting the T-SPOT.TB test conduces to increase the diagnostic rate of TBM.The diagnostic accuracy of the T-SPOT.TB test for CSF may be higher than that for peripheral blood.