Proliferation and Apoptosis of Leukemia Cell Line K562 Treated with HSP90 Inhibitor 17-DMAG / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the role of HSP90 in proliferation and apoptosis of leukemia cells K562 through detecting the effect of HSP90 inhibitors 17-[2-(Dimethylamino) ethyl] amino-17-desmethoxygeldanamycin(17-DMAG) on leukemia K562 cell lines.</p><p><b>METHODS</b>The K562 cells were treated with HSP90 inhibitors 17-DMAG, the semi-quantitative PCR was used to detect HSP90 gene expression, the WST was used to detect the effect 17-DMAG on cell proliferation as well as Annexin V flow cytometry was used to detect the cell apoptosis.</p><p><b>RESULTS</b>After 17-DMAG treated the K562 cells in different stage, the K562 cell growth was obviously inhibited with time dependent (48 h)(r=0.9918) and dose dependent(3.2 µmol/L) manners (r=0.9999) (P<0.01); after the K562 cells in different stage were treated with different concentrations of 17-DMAG, the K562 cells showed significant apoptosis and with dosage-dependent mauner (r=0.9903)(P<0.01); HSP90 mRNA expression decreased significantly after K562 cells were treated with different concentrations of 17-DMAG for 48 hours. 17-DAMG down-regulated the HSP90 mRNA expression in dosage-dependent mauner as well(r=0.9227) (P<0.01).</p><p><b>CONCLUSION</b>HSP90 inhibitor 17-DMAG can inhibit the proliferation of K562 cells and induce their apoptosis. This study result provides laboratory basis for the treatment of leukemia patients with 17-DMAG.</p>

Effect of Heat Shock Protein 90 Inhibitor 17-DMAG on Proliferation and Apoptosis of Acute Lymphocytic Leukemia Cell Line Jurkat / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the effect of heat shock protein 90(HSP90) inhibitor 17-DMAG, an inhibitor specific for heat shock protein 90, on the proliferation and apoptosis of acute lymphocytic leukemia cell lines Jurkat.</p><p><b>METHODS</b>Jurkat cells were collected, then were treated with 17-DMAG. The expression of HSP90 was examined by semi-quantitative RT-PCR analysis, the effect of 17-DMAG on cell proliferation were detected by using WST, and cell apoptosis were detected by using flow cytometry with Annexin V/PI double stenining.</p><p><b>RESULTS</b>After Jurkat cells were treated with different concentrations of 17-DMAG for 48 hours, the HSP90 mRNA expression decreased significantly in dose dependent manner (r=0.9530, P<0.01). The ICwas 3.17 mmol/L when the Jurkat cells were treated with 17-DMAG for 48 h; after treating Jurkat cell with 17-DMAG, the cell proliferation was inhibited(r=0.9903, P< 0.01), the cell apoptosis was increased in dose dependent manner (r=0.9876, P<0.01).</p><p><b>CONCLUSION</b>17-DMAG can inhibit the Jurkat cell proliferation and induce the Jurkat cell apoptosis.</p>

Expression of proBNP and NT-proBNP in Sudden Death of Coronary Heart Disease / 法医学杂志

Objective To study the expression change of pro-brain natriuretic peptide (proBNP) and N-terminal pro-brain natriuretic peptide (NT-proBNP) in sudden death of coronary atherosclerotic heart disease,and to explore its application in forensic diagnosis.Methods Myocardial and blood samples were collected from normal control group,sudden death of coronary atherosclerotic heart disease group and single coronary stenosis group (20 cases in each group).The expression of proBNP in myocardial samples were detected by immunohistochemical staining and Western blotting,and that of BNP mRNA were detected by reverse transcription PCR (RT-PCR).The content of NT-proBNP in plasma were detected by ELISA.Results Immunohistochemical staining showed positive expression of proBNP in both sudden death of coronary atherosclerotic heart disease group and single coronary stenosis group.There was no positive expression in normal control group.For sudden death of coronary atherosclerotic heart disease group and single coronary stenosis group,the relative expression of proBNP protein and BNP mRNA in myocardial tissue and the NT-proBNP content in plasma were higher than that of normal control group (P＜0.05).The NT-proBNP content in plasma of sudden death of coronary atherosclerotic heart disease group was higher than that of single coronary stenosis group (P＜0.05).Conclusion In myocardial ischemia condition,the higher expression of proBNP in cardiac muscle cell shows that the detection of NT-proBNP in plasma can be useful to differentially diagnose the degree of coronary atherosclerotic heart disease and determine whether the sudden death due to coronary atherosclerotic heart disease.

Graphene and Anthraquinone-2-sulfonic Acid Sodium Based Electrochemical Aptasensor for Pb2+ / 分析化学

An electrochemical aptasensor for Pb2+ was constructed based on the layer-by-layer assembly of graphene (GR) and anthraquinone -2-sulfonic acid sodium salt (AQMS) on the surface of Pb2+ aptamer. The mercapto-modified Pb2+ aptamer was first anchored on a gold electrode. Then the highly conductive material of GR was adsorbed on apt through the unique π-π stacking interaction, which was further used for the assembly and signal amplification of the electroactive AQMS. Upon interaction with Pb2+ ions, the apt on the aptasensor undergone conformational switch from a single-stranded form to the G-quadruplex structure, causing the GR assembled with AQMS releasing from the electrode surface into solution. As a result, the electrochemical signal of AQMS on the aptasensor was substantially reduced. Base on this concept, a useful platform for detection of Pb2+ ions was constructed. Under the optimal experimental conditions, the attenuation of peak currents (△Ipa ) showed linear relationship with the logarithm of Pb2+ concentrations (lgCPb2+) over the range from 5. 0×10-10 to 5. 0×10-8 mol/ L. The detection limit was estimated to be 6. 0×10-11 mol/ L.

Study of the regulatory mechanism of EGR-1 expression in HTLV-1 virus Tax protein positive-T cells / 中华微生物学和免疫学杂志

Objective To explore the expression of early growth response gene-1 (EGR-1) in T cells that were positive for Tax protein of human T-cell leukemia virus type 1 (HTLV-1) and its possible reg-ulatory mechanism .Methods A series of expression structures carrying the regulatory elements of EGR-1 in different length and luciferase reporter genes were constructed .TaxP cells were transfected with the con-structs containing reporter genes and cultured with 5μmol/L of NF-κB inhibitor BAY 11-7082 or equal vol-ume of DMSO.After cultured for 24 hours the cells were collected to test the luciferase activity .BAY 11-7082 or equal volume of DMSO was added into the supernatant of TaxP cell culture to test the expression of EGR-1 protein by Western blot after 24 hours of culture .Tax and its mutants M22 and M47 were transfected into 293 T cells respectively to test the expression of EGR-1 protein by Western blot after 24 hours of culture . Results The expression structures carrying the regulatory elements of EGR-1 in different length and their mutants followed by luciferase reporter genes were successfully constructed .The luciferase activity in the cells transfected with the constructs containing the elements E 1 and E2 were higher than that transfected with E3, DelE and MutE, but the reporter gene expressions were decreased with the interference of BAY 11-7082 (P<0.01).However, there were no significant changes with the luciferase activity in the cells transfected by elements E3, DelE and MutE.Western blot analysis indicated that the expression of EGR-1 protein was significantly decreased with the interference of BAY 10-7082 .The expression of EGR-1 protein in M22 mu-tants-transfected 293 T cells were decreased significantly in comparison with those by wild type tax-and M47-transfected cells .Conclusion NF-κB was the key nuclear factor in regulating the expression of EGR-1 pro-tein in Tax-positive T cells .

Risk factors analysis of postsurgical gastroparesis syndrome and its impact on the survival of gastric cancer after subtotal gastrectomy / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the risk factors of postsurgical gastroparesis syndrome (PGS) after subtotal gastrectomy in gastric cancer and the impact of PGS on prognosis.</p><p><b>METHODS</b>Clinical data of 422 patients who underwent subtotal gastrectomy for gastric cancer in the Central Hospital of Huzhou Sity from January 2004 to May 2010 were analyzed retrospectively. Risk factors of PGS were indentified and the recurrence-free survival was compared between the patients with and without PGS.</p><p><b>RESULTS</b>PGS occurred in 42 patients (9.5%). Univariate analysis showed that: age over 65, combination of anxiety disorder, low-albuminemia in perioperative period, pyloric obstruction in preoperative period, high serume glucose level (≥ 11.2 mmol/L) in postoperative period, Billroth II (gastroenterostomy, operation time over 4 hours, using patient-controlled analgesia, or intravenous fluid over 3500 ml/d (all P<0.05) were prone to develop PGS. These might be potential clinical risk factors associated to PGS. Correlation analysis showed the number of clinical risk factors was positively correlated with the incidence of PGS (r=0.967, P<0.05). A total of 215 cases (50.9%) were followed up for 3-60 months. The mean recurrence-free survival time of patients with PGS was 26.1 months, which was shorter than that of those without PGS (33.4 months, P=0.029).</p><p><b>CONCLUSIONS</b>Gastric cancer patients with the clinical risk factors mentioned above are prone to develop PGS after subtotal gastrectomy. PGS is associated with poor prognosis.</p>

Chi-square analysis of the reduction of ATP levels in L-02 hepatocytes by hexavalent chromium

This study explored the reduction of adenosine triphosphate (ATP) levels in L-02 hepatocytes by hexavalent chromium (Cr(VI)) using chi-square analysis. Cells were treated with 2, 4, 8, 16, or 32 μM Cr(VI) for 12, 24, or 36 h. Methyl thiazolyl tetrazolium (MTT) experiments and measurements of intracellular ATP levels were performed by spectrophotometry or bioluminescence assays following Cr(VI) treatment. The chi-square test was used to determine the difference between cell survival rate and ATP levels. For the chi-square analysis, the results of the MTT or ATP experiments were transformed into a relative ratio with respect to the control (%). The relative ATP levels increased at 12 h, decreased at 24 h, and increased slightly again at 36 h following 4, 8, 16, 32 μM Cr(VI) treatment, corresponding to a "V-shaped" curve. Furthermore, the results of the chi-square analysis demonstrated a significant difference of the ATP level in the 32-μM Cr(VI) group (P < 0.05). The results suggest that the chi-square test can be applied to analyze the interference effects of Cr(VI) on ATP levels in L-02 hepatocytes. The decreased ATP levels at 24 h indicated disruption of mitochondrial energy metabolism and the slight increase of ATP levels at 36 h indicated partial recovery of mitochondrial function or activated glycolysis in L-02 hepatocytes.

Influence of extracellular HMGB1 on the virus replication of HTLV-1 infected T cells / 中华微生物学和免疫学杂志

Objective To investigate the influence of extracellular high mobility group box 1 (HMGB1) on viral replication in HTLV-1 infected T cells.Methods HMGB1 in culture supernatants of adult T-cell leukemia virus 1 (HTLV-1) virus-negative cell:Jurkat,MOLT4 cells and HTLV-1 virus-positive cells:MT2,MT4,was detected by ELISA;The HTLV-1 long terminal repeat reporter gene (pHTLV-1-LTR-luc) was transfected into MT2 cells by Tfx-50-mediated transfection,and 0.25,0.50,0.75 μg/ml of HMGB1 polyclonal antibody(HMGB1 PcAb) and its isotype control rabbit IgG antibodies,0.03,0.1,0.3 μg/ml rhHMGB1 and its control PBS,were added into culture supernatant respectively,then luciferase activity was detected after 48 h;Similarly,0.25 μg/ml HMGB1 PcAb and the isotype control antibody,0.3 μg/ml rhH-MGB1 and the control PBS were added to the culture supernatant of MT2 cell,the viral gene,pol1,pol2,gag,env,etc,were performed by real-time PCR.Results Culture supernatant HMGB1 levels has no significant difference between HTLV-1 positive cells MT2 and MT4 and the other two virus-negative T cell lines;Compared with isotype control antibody group,the culture supernatant,to which is added 0.25 μg/ml HMGB1 PcAb,can significantly inhibit the HTLV-1-LTR transcriptional activity and suppress the expressions of the viral gene pol1,pol2,gag,env.Compared with the control PBS,0.3 μg/ml rhHMGB1 significantly promotes the transcriptional activity of the HTLV-1-LTR and the expressions of the viral gene pol1,pol2,gag,env.Conclusion The extracellular HMGB1 can promote viral replication of HTLV-1 infected T cells.

Expression of FHIT, FN and PTEN in hepatocellular carcinomas / 中华普通外科杂志

Objective To investigate the expression of fragile histidine triad(FHIT),fibronectin (FN) and phosphatase and tensin homology deleted on chromosome ten( PTEN )in human hepatocellular carcinomas (HCC) and their relationship with pathological characteristics and prognosis of HCC.Methods Immunohistochemistry was used to detect expression of FHIT,FN and PTEN in cancerous tissues from 138 HCC patients.The correlation between their expression and clinicpathological features and prognosis were analyzed.Results FHIT,FN,PTEN protein expressed differently between HCC and adjacent mucosa ( respectively x2 =5.968,7.380,4.962,all P ＜ 0.05 ),and the expression level was different with tumor size,tumor number,grades and tumor lymph node metastasis( P ＜ 0.05 ).In FHIT and FN positive group,the recurrence free survival rates were lower than those in negative groups( respectively x2 =4.443,9.867,all P ＜0.05),while in PTEN positive group patients' recurrence free survival rates were higher than those with PTEN negative expression ( x2 =4.199,P ＜ 0.05 ).Conclusions FHIT,FN,PTEN were abnormally expressed in HCC.Positive expression of FHIT and FN predicts poor prognosis,while positive PTEN indicates fair prognosis.

Study on the intelligence quotient characteristics of mild iodine deficiency disorders children and sociocultural condition abnormal children / 中国地方病学杂志

Objective To investigate the intelligence standard for diagnose the sub-cretin children and children with mental retardation of socio-cultural type.Methods The full intelligence quotient(IQ),verbal intelligence quotient(VIQ)and performance intelligence quotient(PIQ)was tested by Wechsler scale(C-WISC)for mild iodine deficiency disordem children,children living in abnormal socio-cultural condition and normal children aged 7～14 years old in Qinba mountain area.The test results had been compared between the groups.Results There were no significant difference between psychomotor functioning well children and children living normal sociocuhural condition in VIQ,PIQ and full IQ(89.24±18.44 vs 90.75±17.58,87.58±15.78 vs 88.95±15.56,87.42±17.84 vs 89.02±17.18,t=1.14,1.19 and 1.24,respectively,all P＞O.05).PIQ and full IQ were significantly lower in mild iodine deficiency disorders children than in children with abnormal socio-cultural background (65.81±10.22 vs 72.33±13.23,62.42±12.31 vs 68.13±14.54,t=3.26,2.55,P＜0.01 or＜0.05,respectively).But the VIQ was not significantly different between these two groups.The average difference of VIQ and PIQ among mild iodine deficiency disorders children wag-0.32 without significant difierence(t=0.28,P＞0.05),however it was-2.91 among children under abnormal socio-cultural condition with significant difierenee(t=-3.59,P＜0.01).Conclusions IQ for iodine deficiency disorders children is characterized by that VIQ is damaged in parallel with PIQ,while that in children under abnormal soeio-cuhural condition is marked by that VIQ is retarded more severely than PIQ,which ean be used as an intelligence standard for differentiating the sub-cretin children from children wjth socio-cuhural mental retardation.

Experimental Study of Antitoxic Effect of Sodium Selenite on Guinea Pig Cochlea Injury Induced by Gentamicin in Vivo / 中国康复理论与实践

@#ObjectiveTo explore the effect of sodium selenite on gentamicin-induced ototoxicity.MethodsTwenty guinea pigs were randomly divided into the experimental group and control group with 10 animals in each group. The animals were treated with i.m injection of gentamicin (200 mg/kg/d for 8 consecutive days, and at same time, the experimental group was added with p.o tablet of sodium selenite (1 mg/kg/d). Before and after the administration, hearing function was evaluated by examination for auditory brain stem responses (ABR). The cochlear outer haircells were observed by scanning electronic microscope (SEM) and transmission electronic microscope (TEM).ResultsABR threshold of the control group was about 30 dB above that of the experimental group (P<0.01). The latency of waveⅠof the control group was about 0.23 ms above that of the experimental group (P<0.01). Under SEM, the cilia of the majority outer hair cells of the control group lodged even disappeared, while that of the experimental remained regular. Under TEM, in outer hair cells of the control group, mitochondrial crests were obscure, out-membrane was damaged and local protruding, the number of secondary lysosomes was increased, myeloid bodies appeared, but in the experimental group, outer hair cells basically remained normal.ConclusionSodium selenite has antitoxic effect on guinea pig cochlea injury induced by gentamicin in vivo.

Study on sexual behavior and HIV/STIs among miners in Yunnan province / 中华流行病学杂志

<p><b>OBJECTIVE</b>This paper aims to describe human immunodeficiency virus/sexual transmitted infections (HIV/STIs) related knowledge, attitudes, practice and the prevalence of HIV/STIs amongst miners.</p><p><b>METHODS</b>Two focus-group related discussions with a total number of 13 members including Community Advisory Boards (CAB) and 12 miners were conducted in a mining township in Yunnan province. Questionnaire surveys and HIV/STIs tests were conducted among 233 miners recruited by cluster sampling in two towns where the mines were located.</p><p><b>RESULTS</b>The average age of respondents was 28 year old with 82.8% of them younger than 35 year old. 95.3% of the respondents attended the education level of junior middle school. AIDS related knowledge among miners was low. The percentage of right answers to the routes of transmission was only 54.4%. The ratio of self-reported prostitutes visits was 9.0%. The prevalence rates of Neisseria gonrrhoeae, HIV and Chlamydia trachomatis were 0.4%, 0.4%and 8.2% respectively. The correlation between Chlamydia trachomatis infection and education (P = 0.0347) was significant, and so was that between Chlamydia trachomatis infection and marriage status (P = 0.032).</p><p><b>CONCLUSION</b>This study showed that the awareness of HIV/STIs prevention was limited and the rate of condom use was low, suggesting that miners needed to be viewed as a key population in HIV/STIs prevention and control.</p>

Protein kinase C is partly involved in c-fos protein expression of nocuously-activated neurons but may not in concomitant modulatory action through opioid receptors at the spinal level in rats / 生理学报

The present study was aimed to examine if protein kinase C (PKC) activation is necessarily involved in both the c-fos protein expression in the nocuously-activated c-fos protein-like immunoreactive (Fos-LI) neurons and the concomitant opioid receptor-mediated modulation in the dorsal horn circuitry of the spinal cord. Formalin was injected into a hindpaw of rats 5 min after the rats were pretreated with intrathecal (i.t.) administration of chelerythrine (Chel), an inhibitor of PKC, naloxone (Nal), combined administration of these two (Chel + Nal), or vehicle (n=5 in each group),respectively. By using immunocytochemical techniques, the formalin-induced Fos-LI neurons in the lumbar dorsal horn were calculated 1 h after formalin injection. The results showed that: (1) i.t. Chel significantly reduced the number of Fos-LI neurons in the dorsal horn of the spinal cord on the side ipsilateral to the formalin injection, showing a decrease by 60.3% (P<0.001) as compared to that observed in the i.t.vehicle group; (2) i.t. Nal significantly increased the number of Fos-LI neurons in the ipsilateral dorsal horn, with an increase of 46.0% (P<0.01) as compared to that in the i.t.vehicle group, the highest percentage increase being found in the deeper laminae of the dorsal horn; and (3) i.t. Chel + Nal also exhibited a significant decrease in Fos-LI neurons in the ipsilateral dorsal horn as compared to i.t. Nal group, showing a reduction of 53.2%, a value similar to that in the i.t. Chel group. These results suggest that: (1) PKC plays a role in the c-fos protein expression only in nearly one half of the Fos-LI neurons in the dorsal horn; and (2) PKC is possibly not involved in the concomitant modulation on the nociception mediated by micro- (and also partly delta-) opioid receptors in the spinal cord.