AIM: To elucidate the effect of ginsenoside Rb1 (Gs-Rb1) on the glucose metabolism to improve the viability of the cardiomyocytes under hypoxia, and whether hypoxia-inducible factor 1α(HIF-1α) and/or AMPKαare involved in the process.METHODS: The neonatal rat cardiomyocytes were cultured, and randomly divided into control group, hypoxia (1% O2 , 94% N2 and 5% CO2 ) group, Gs-Rb1 (200 μmol/L) group, Ara-A (500 μmol/L) group, Gs-Rb1 +Ara-A group, YC-1 (5 μmol/L) group, Gs-Rb1 +YC-1 group, Ara-A +YC-1 group and Gs-Rb1 +YC-1 +Ara-A group.After the intervention for 8 h, the cell viability was analyzed by MTT assay.The protein levels of AMPK, HIF-1αand glucose transporter-4 (GLUT-4) were determined by Western blot.The activities of heterophosphatase (HK), phos-phofructokinase (PFK) and lactic dehydrogenase (LDH) were measured by ELISA.RESULTS: Gs-Rb1 significantly im-proved the viability of hypoxic cardiomyocytes, which was significantly inhibited by YC-1 and Ara-A.In addition, YC-1 and Ara-A had a synergistic effect.Gs-Rb1 increased the protein levels of AMPK and HIF-1αin the hypoxic cardiomyo-cytes, which was significantly inhibited by Ara-A and YC-1.Gs-Rb1 significantly increased the expression of GLUT-4 on the cytomembrane of hypoxic cardiomyocytes, which was significantly inhibited by YC-1 or Ara-A, especially Ara-A +YC-1.Gs-Rb1 significantly increased the activities of HK, PFK and LDH, all those were significantly inhibited by YC-1 or Ara-A.Besides, YC-1 and Ara-A had a synergistic effect.CONCLUSION: Gs-Rb1 improves the viability of hypoxic car-diomyocytes, which may be related to the regulation of glucose uptake and enhancement of glycolysis by synergy of both
HIF-1αand AMPK.

A 73-year-old man developed a skin-colored granule-sized papule on the scrotum 10 years prior to the presentation,which had increased in size and number without obvious symptoms.Histopathological examination showed multiple enlarged intradermal cysts.The cyst wall consisted of one or two layers of flat or columnar cells.Decapitation secretion was seen in the inner layer cells,and the out layer of cysts was formed by flat or cubic myoepithelial cells.A diagnosis of apocrine hidrocystoma of the scrotum is made.

Objective To investigate the effects of empathy on the insight of first-episode schizophrenia.Methods Empathy with tbe interpersonal reactivity index-C (IRI-C) was applied to 71 patients with first-episode schizophrenia and 69 healthy controls.In addition,schizophrenic patients were evaluated with scale of unawareness of mental disorder (SUMD) for the scores of clinical insight and positive and negative syndrome scale (PANSS) for clinical symptoms.Results Compared with the control group,the schizophrenic patients showed lower scores in the perspective taking ((13.85±1.86) vs (6.74±2.56),P＜0.01),empathic concern ((14.28±2.16) vs (17.49± 2.73),P＜0.01) of IRI-C,but higher subscale score ((10.28±2.78) vs (8.93±1.93),P＜0.01) in personal distress.Pearson correlation analysis showed that,there were negatively significant correlations between insight score and perspective taking (r=-0.429,P＜0.01),empathic concern (r=-0.709,P＜0.01) subscores of IRI-C respectively.In addition,the insight total score was positively correlated with the positive symptoms subscores (r=0.545,P＜0.01) of PANSS.Conclusion Patients with first-episode schizophrenia have significant empathy defects.And the empathic deficits are associated with the insight impairment among the schizophrenic patients,suggesting that patients' insight level can change by improving their empathy ability.

Objective To explore the physiological and psychosocial risk factors of the patients with somatoform disorders and to provide references for clinical psychological intervention.Methods Totally 60 patients with somatoform disorder and 60 normal controls were tested with biochemical analyzer,electrocardiogram machine,electroencephalogram machine,computed tomographic scanner,general situation questionnaire,minnesota multiphasic personality inventory(MMPI),toronto alexithymia scale(TAS),life event scale(LES),defense style questionnaire(DSQ) and social support rating scale(SSRS) to explore the physiological and psychosocial risk factors of the patients with somatoform disorders.Results The score of immature defense styles of the study group(4.04±0.89) were higher than those in the control group(3.43 ± 0.65)with significant difference (P＜0.05).The total and factor scores of TAS of study group((86.20±11.15),(21.12±3.28),(26.08±3.86),(18.21±4.69),(23.44±5.60)) were higher than control group((65.00±10.12),(12.43±5.18),(15.64±5.57),(15.56±3.16),(18.71 ±4.30)) with significant difference (P＜0.05).The MMPI scale scores of Hs (hypochondriasis),D (depression),Hy (hysteria),Men-Nancy (Mf),Paranoid (Pa),Psychotic (Pt) were higher in patients with somatoform disorder ((72.79± 10.50),(68.46±13.63),(79.03±12.12),(51.72±7.74),(57.98±10.60),(54.98±8.83)) than those in normal controls((51.66±10.11),(47.96±9.42),(51.47±11.90),(43.66±8.77),(47.67±6.69),(47.48±8.67)) with significant difference(P＜0.05).The total life event score and the negative life event score of LES of the study group ((89.66±68.60),(82.65±65.16)) were higher than those in the control group((56.00±56.79),(39.14± 46.05)) with significant difference (P＜ 0.05).Multiple factors logistic regression analysis showed immature defense styles,higher alexithymia score,higher negative life event score,higher HS,D,Hy of MMPI may increase the risk of somatoform disorders(OR=6.84,9.12,5.83,2.69,1.81,11.20,P＜0.01).Conclusion Patients with somatoform disorder are almost in normal physiological condition.They have the personality characteristics and significant alexithymia.Their social support are almost normal,but they have more negative life events and their defense styles are almost immature.

Objective Through constructing prokaryotic expression vector pET-30a-GPI-B7-1, to gain purified GPI-B7-1 fusion protein so as to confirm the tumor immune effect. Methods The DNA fragment encoding the signal for GPI-anchor attachment of hPLAP-1 and the cDNA encoding the human costimulatory molecule CD80 ( BT-1 ) were cloned from fresh placenta and human peripheral blood monocytes (PBMC) respectively. The two fragment were annealed to form a fusion gene (GPI-BT-1) by PCR. Then the fusion gene was inserted into the prokaryotic expression vector pET-30a, resulting in pET-30a-GPI-BT-1. Transfer to E. coli BL21, purified fusion protein were analysed by SDS-PAGE and Western blot. Results Agarose gel electrophoresis map of GPI and BT-1 PCR products show that GPI goal gene strap was seen at 133bp region and BT-1 goal gene strap at 792 region. Identification of recombinant pET-30a-GPI-B7-1 by restriction enzyme and PCR illustrate two goal fragment for 5000 bp and 900 bp, to realize the expression of fusion gene ( GPI-B7-1 ) at the E. coli BL21. The fusion protein was successfully produced in the pET expression system induced by IPTG and purified by Ni2 + -NTA agarose column. By SDS-PAGE and Western blot analysis, the observed molecular weight of the fusion protein was 38 kDa. Conclusion The purified GPI-B7-1 fusion protein can be obtained from E. coli BL21 transfered by prokaryotic expression vector pET-30a-GPI-B7-1, which will prove useful tool for the study of tumor immune therapy.

Objective To investigate the clinical value of sB7-H3 in predicting the severity of acute pancreatitis at early stage. Methods By using the double antibody sandwich ELISA method, the level of plasma sB7-H3 was measured at 24h after onset of abdominal pain in 75 patients with acute pancreatitis (MAP30, MSAP20, SAP25), and 20 healthy persons were enrolled in the controlgroup.The sensitivity and specificity correlations of sB7-H3 in acute pancreatitis with severity degree , as well as with the clinical detection index , were also evaluated. Results The level of plasma sB7-H3 at 24 h in the AP group was significantly higher than that in the healthy control ( HC) group (t = 3.925, P = 0.0002), however, no significant difference was found between the MAP groep and the HC group (P>0.05). The levels of plasma sB7-H3 in the MSAP and the SAP group were significantly higher than that in the HC group (P<0.05和P<0.001)or the MAP group (P<0.05 和P<0.01);The level of plasma sB7-H3 in the SAP group was also markedly higher than that in the MSAP group (P < 0.01). sB7-H3 had a linear positive correlation with LDH、hs-CRP、WBC(P<0.05). ALB had a linear negative correlation with and Ca (S)(P<0.05). By the cutoff of sB7-H3, the sensitivity and specificity to judgethe above moderate pancreatitis were 88.9%and 83.3%,and to judgethe SAP were 96%and 96%. Conclusion sB7-H3 has important clinical value to judge the severity of acute pancreatitis at early time with high sensitivity and specificity , with a linear correlation with the clinical severity index.

Aim ToinvestigatetheeffectsofCa2+/calmodulin-dependent protein kinase Ⅱ inhibitor KN-93 on calcium overload-induced heart injury.Methods Thirty-twoisolatedratheartswererandomlydivided into the control group,KN-93 control group,calcium paradox group,and calcium paradox with KN-93 treat-ment group.Left ventricular pressure were recorded, and the heart function was evaluated by the left ventric-ular end-diastolic pressure (LVEDP ) and developed pressure (LVDP).Coronary flow (CF)were collect-ed,and lactate dehydrogenase (LDH)content was de-termined.Triphenyltetrazolium chloride staining was usedtomeasuretheinfarctsize.Results Compared with the control group,KN-93 at 2. 5 μmol·L-1 had no effects on coronary flow,cardiac performance and cell death at the end of perfusion in normal rats (P>0. 05 );The hearts of calcium paradox exhibited a de-crease in LVDP and CF,meanwhile an increase in LV-EDP,LDH,and infarct size of 18 ±7. 2% (P <0. 01).2. 5 μmol·L-1 KN-93 further increased the levels of LVEDP,LDH and infarct size (P<0. 01)in Ca2+paradoxical hearts,while it provoked the decline intheCFandLVDP(P<0.01).Conclusion The data demonstrates that KN-93 aggravates heart injury in calcium paradox,it also suggests that CaMKⅡ is in-volved in the Ca2+overload-induced heart injury.

Objective To investigate the quality of concentrated platelet with less WBC by improved white membrane method,so as to provide the basis for preparation of the concentrated platelet in the future.Methods Testing the quality indicators of 56 cases of concentrated platelet with less white blood cells by improved white membrane method,and comparing the quality indicators of 56 cases of collected platelet at random by machine.Testing the expression rate of platelet membrane surface glycoprotein molecules CD62P and PAC-1 before and after filtering in different storage time,and comparing the expression rate of 37 cases of collected platelets at random by machine.Results The differences of quality indicators between concentrated platelet with less white blood cells by improved white membrane methods and collected platelet at random by machine were not statistically significant (P＞0.05),such as volume,platelet content,Ph,amount of RBC mixed within.But The differences of amount of WBC mixed with were statistically significant (P＜ 0.01),both of sterility tests were negative.The differences of the expression rate of platelet membrane surface glycoprotein molecules CD62P and PAC-1 during storage period within 3 days were not statistically significant (P＞0.05).Conclusion The various quality indicators of concentrated platelet with less white blood cells by improved white membrane methods attain mixed concentrated platelet and platelet national standards,and in vitro platelet activity kept well.Platelet special leucocyte filter does not cause obvious platelet activation and damage within three days of storage period of concentrated platelet in filter processing.Collected platelet by machine is being activated and damaged continuously when kept in platelet oscillation device 22±2℃ for 3 days,but activation and damage does not significantly change the platelet activity in vitro.

Objective To investigate the effects of four tyrosine metabolites on the proliferation, cell cycle and chemosensitivity of lung cancer cells.Methods A549 cells were cultured with different concentrations of tyrosine metabolites:tyrosine, 4-hydroxyphenylpyruvate(HPPA), homogentisic acid(HGA) and fumaric acid (FA) in vitro.High content imaging analysis system was used to monitor cell proliferation.Flow cytometry was used to detect cell cycle and apoptosis.Protein and mRNA level of Bax and Bcl-2 were measured by Western blot.Results Tyrosine metabolites had no effect on cell cycle and proliferation of A549 cells.HGA decreased the cell apoptosis of A549 cells induced by Gefitinib with depressed Bax expression and elevated Bcl-2 expression.However, FA increased the apoptosis of A549 cells induced by Gefitinib with up-regulation of Bax and down-regulation of Bcl-2.No obvious effect of tyrosine and HPPA was detected on Gefitinib induced cell apoptosis.Conclusion HGA and FA may play an important role in drug sensitivity of lung cancer cells, indicating a critical role of tyrosine metabolism in lung cancer.

OBJECTIVETo construct a GFP/Puro double-labeled lentiviral expression vector for CK8 silencing and assess the effects of CK8 silencing on cell apoptosis.

METHODSThe siRNA sequences of CK8 were inserted into the lentiviral expression vector GV248 and transfected into 293T cells with the packaging plasmids PMD and SPA. The lentivirus was collected at 24 and 36 h post-transfection. Flow cytometry was used to detect the virus titer and the positive cells were selected with puromycin. The knockdown of CK8 was examined by Western blotting. The effect of CK8 down-regulation on cell apoptosis induced by cisplatin was detected with Annexin V/PI staining.

RESULTS AND CONCLUSIONWe successfully constructed CK8 interference lentiviral vector and obtained a stable cell line with CK8 knock-down that was sensitive to cisplatin-induced apoptosis.

Apoptosis ; Cell Line ; Down-Regulation ; Genetic Vectors ; Humans ; Keratin-8 ; genetics ; Lentivirus ; Plasmids ; RNA Interference ; RNA, Small Interfering ; Transfection