OBJECTIVE:To study the effects of Acivicin or/and cisplatin on the apoptpsis of HepG2 and expression of Bcl-2,c-myc and p53.METHODS:Acivicin and cisplatin were used to treat cultured HepG2 cell line separately or in combination.The cytotoxicity was measured by MTT assay,the apoptosis by TUNEL assay and relevant regulator genes by immunohistochemical SP method.RESULTS:Acivicin in concentrations of 0.28,0.56,0.84,1.12 and 1.40mmol/L,the apoptosis rates of HepG2 were (3?1.25)%,(3.7?2.0)%,(10?1.25)%,(17.4?4.5)%and (16.9?3.5)%respectively.Acivicin in concentration of 1.4mmol/L,the apoptosis rates at 24,48 and 72h were(16.9?3.5)%,(27.9?4.3)%and (47.2?3.0)%respectively,which was significantly different from control group.The apoptosis rate of HepG2 induced by cisplatin(67?mol/L)was (73.4?1.5)%at 24h.The apoptosis rate of HepG2 induced by Acivicin(1.4mmol/L)plus cisplatin(67?mol/L)was(94.7?0.5)%at 24h,which was markedly higher than those induced by Acivicin or cisplatin alone.The expressions of Bcl-2,c-myc and p53 in all groups were increased comparing with those in control group.CONCLUSION:This study indicates that the apoptosis of HepG2 induced by Acivicin was in a dose-dependent and time-dependent manner.When Acivicin and CDDP used simultaneously,the effect of CDDP on HepG2 apoptosis was enhanced greatly.

Objective To investigate the influence of hepatitis B virus X (HBx) protein on the activity of NLRP3 inflammasome as well as the mechanism of accelerating HBV-related hepatocellular carcinoma (HCC).Methods The HepG2 cell strains were divided into the 5 groups:blank control group (without plasmid transfection),empty vector group [transfected with pE green fluorescent protein (GFP)-N1 vector plasmid],fulllength HBx protein group (transfected with pEGFP-N1-X plasmid),HBx1-127 group (transfected with pEGFP-N1-X1-127 plasmid),HBx1-101 group (transfected with pEGFP-N1-X1-101 plasmid).(1) The expressions of HBx protein and NLRP3 inflammasome were detected by Western blot [Lipopolysaccharide (LPS) + ATP intervention was performed in the blank control group].(2) The HepG2 cells in the full-length HBx protein group were respectively intervened by glibenclamide and ammonium pyrrolidinedithiocarbamate (APDC),and the expressions of IL-1β and IL-18 were detected by enzyme-linked immunosorbent assay (ELISA).(3) The expressions of reactive oxygen were detected by flow cytometry.The measurement data with normal distribution were presented by (x) ± s.The one-way ANOVA was adopted in the comparison among groups while the t test was used in the pairwise comparison.Results (1) The results of Western blot showed:① the relative expressions of HBx recombinant plasmid fusion protein inside the HepG2 cells in the blank control group,empty vector group,full-length HBx protein group,HBx1-127 group and HBx1-101 group were 0.07 ±0.03,0.92 ±0.13,0.84 ±0.11,0.30 ±0.06 and 0.29 ± 0.05,respectively.The expressions in the HBx1-127 group and the HBx1-101 group represented the expressions of HBx1-127 protein and HBx1-101 protein.There were statistically significant differences among the 5 groups (F =61.790,P ＜ 0.05).The relative expression of full-length HBx protein group was significantly different from that of blank control group,HBx1-127 group and HBx1-101 group (t =12.070,7.465,7.801,P ＜0.05).There was no statistically significant difference between full-length HBx protein group and empty vector group (t =0.867,P ＞0.05) and between the HBx1-127group and HBx1-101 group (t =0.146,P＞0.05).② The relative expression of NLRP3 inflammasome protein inside the HepG2 cells in the blank control group,full-length HBx protein group,HBx1-127 group,HBx1-101 group and LPS + ATP group were 0.29 ±0.06,0.83 ±0.14,0.27 ±0.06,0.27 ± 0.05 and 0.90 ± 0.16,respectively,with a statistically significant difference among the 5 groups (F =29.550,P ＜ 0.05).The relative expression of NLRP3 inflammasome protein of LPS + ATP group was significantly different from that of blank control group,HBx1-127 group and HBx1-101 group,respectively (t =6.310,6.565,6.741,P ＜0.05).There were statistically significant differences between the full-length HBx group and the HBx1-127 group or HBx1-101 group (t =6.381,6.584,P ＜ 0.05) and no statistically significant difference between LPS + ATP group and full-length HBx protein group (t =0.580,P ＞ 0.05).(2) The results of ELISA showed:①) the expression of IL-1β inside the HepG2 cells in the blank control group,full-length HBx protein group,HBx1-127 group,HBx1-101 group and LPS + ATP group was (87 ± 9)pg/mL,(587 ±56)pg/mL,(125 ±12) pg/mL,(113 ± 13) pg/mL and (677 ± 74) pg/mL,respectively,with a statistically significant difference among the 5 groups (F =139.010,P ＜ 0.05).The expression of IL-1 β of LPS + ATP group was significantly different from that of blank control group,HBx1-127 group and HBx1-101 group (t =13.691,12.752,13.001,P ＜0.05).The expression of IL-1β of full-length HBx group was significantly different from that of the HBx1-127 group and the HBx1-101 group (t =14.051,14.283,P ＜ 0.05).There was no statistically significant difference between the LPS + ATP group and the full-length HBx protein group (t =1.691,P ＞0.05).The expression of IL-18 in the blank control group,full-length HBx protein group,HBx1-127 group,HBx1-101 group and LPS + ATP group was (43 ±8)pg/mL,(252 ±38)pg/mL,(70 ± 13)pg/mL,(63 ± 10)pg/mL and (263 ±48)pg/mL,respectively,with a statistically significant difference among the 5 groups (F =44.010,P ＜0.05).The expression of IL-18 of LPS + ATP group was significantly different from that of blank control group,HBx1-127 group and HBx1-101 group,respectively (t =7.848,6.722,7.065,P ＜ 0.05).The expression of IL-18 of full-length HBx group was significantly different from that of HBx1-127 group and HBx1-101 group (t =7.882,8.331,P ＜ 0.05).There was no statistically significant difference between LPS + ATP group and full-length HBx group (t =0.326,P ＞ 0.05).②The expressions of IL-1β and IL-18 in the HepG2 cells of the full-length HBx protein were (587 ± 91)pg/mL and (243 ± 22) pg/mL before the addition of glibenclamide,(115 ± 17) pg/mL and (90 ± 12) pg/mL after the addition of glibenclamide,respectively,with statistically significant differences before and after the addition of glibenclamide (t =8.800,10.566,P ＜ 0.05).The expressions of IL-1β and IL-18 in the HepG2 cells of the full-length HBx protein were (573 ± 89) pg/mL and (252 ± 24) pg/mL before the addition of APDC,(124 ±21)pg/mL and (116 ± 15)pg/mL after the addition of APDC,respectively,with statistically significant differences before and after the addition of APDC (t =8.516,8.269,P ＜ 0.05).(3) The results of flow cytometry showed that the relative expression of reactive oxygen in the HepG2 cells in blank control group,fulllength HBx protein group and LPS + ATP group were 66 ± 14,275 ± 54 and 388 ± 88,with statistically significant differences among the 3 groups (F =22.130,P ＜ 0.05) and between the full-length HBx protein group or LPS +ATP group and blank control group (t =6.489,6.256,P ＜ 0.05).There was no statistically significant difference between full-length HBx protein group and LPS + ATP group (t =1.887,P ＞ 0.05).Conclusion HBx protein may play an important role in the occurrence and development of HBV-related HCC by activating NLRP3 inflammasome through inducing reactive oxygen generation in the HepG2 cells.

Objective To analyze effects of maternal passive smoking at different exposure level in different period of pregnancy on fetal birth weight and to reveal the influence of different measurement methods on the effects of maternal passive smoking.Methods Meta analysis was applied.NCBI,OVIDC-MEDLINE,CNKI,VIP and CBM were searched,all searched studies were retrieved,and their references were checked for other relevant publications,with the language limited to Chinese and English.The search was finished by April 2008.Pooled mean differences with 95% confidence interval were estimated using data extracted from papers.Results A total of 38 papers were searched,and 19 of them were prospective studies and 19 were retrospective studies,respectively.Maternal passive smoking was associated with a reduction of 67.62 g(95% CI:-90.15～-45.09)in mean birth-weight,and the adjusted reduction was 44.92 g(95% CI:-67.07～-22.77).Based on biochemical markers measurement,the pooled effect size was-73.87 g(95% CI:-113.41～-34.34),based on self-report measurement,the pooled effect size was-62.93 g(95% CI:-84.49～-41.37).The lowest and highest level exposure's effect sizes were-44.61 g(95% CI:-78.36～-10.87)and-116.37 g(95% CI:-180.74～-52.01),respectively.Maternal passive smoking in early pregnancy can reduce the birth-weight by 2.70 g(95% CI:-37.74～32.33),however,it was not statistically significant.Conclusions Maternal passive smoking during pregnancy could reduce infant birth-weight.The middle or late period of pregnancy might be the sensitive period for passive smoking's effect.There is no clear threshold value for passive smoking causing low birth weight.

Objective To study the clinical effect of escitalopram oxalate in treating patients with depression accompanied with anxiety disorder.Methods Sixty-four patients with depression accompanied with anxiety disorder (met DSM-IV diagnostic criteria)were randomly divided into study group (n =32)who were treated with escitalo-pram oxalate 5-20mg/d and control group (n =32)with mirtazapine 15-45 for eight weeks.Clinical effect was evalua-ted with the Hamilton Depression Scale (HAMD)and the Hamilton Anxiety Scale (HAMA)as well as the adverse reactions with the Treatment Emergent Symptom Scale (TESS).Results There were no statistically significant differ-ences (P >0.05)in HAMD and HAMA scores between the study group and control group before treatment,but after treatment for 8 weeks,HAMD score showed statistically significant difference (P =0.000)and the difference of HAMA score was statistically significant (P =0.010).The total effective rate of the study group was 93.8%,which of the control group was 87.5%,the clinical effect had no statistically significant difference (P =0.391 ).The study group and the control group had equal effect,and there were no serious adverse reactions.Conclusion Escitalopram oxalate has high anti-depressant and anxiolytic effect,fewer adverse reactions,good tolerability and compliance,there-fore,escitalopram oxalate can be popularized in treating patients with depression accompanied with anxiety disorder.

OBJECTIVE： To determine the gentamycin sulphate,ephedrine hydrochloride and dexamethasone sodium phosphate in nasal drops without need of separation.METHODS： Gentamycin was determined via the dihydrolutidine derivatives produced by Hantzsh reaction using UV-spectrophotometry.The detecting wavelengths was 330nm.A dual-wavelength spectrophotometry was used for determination of ephedrine hydrochloride and dexamethasone sodium phosphate.The detecting wavelengths were 256.5nm and 241.6nm and the reference wavelengths were 228.4nm and 266.4nm for ephedrine hydrochloride and dexamethasone sodium phosphate,respectively.RESULTS： The average recovery rates of gentamycin sulphate,ephedrine hydrochloride and dexamethasone sodium phosphate were 100.74％ (CV=0.2％ ,r=0.9 999,n=5),100.15％ (CV=0.66％ ,r=0.9 997,n=5)and 99.46％ (CV=0.35,r=0.9 996,n=5)respectively.CONCLUSION： This method is simple,rapid,accurate and stable and suitable for rapid quality control of compound gentamycin sulphate nasal drops.

Objective:To investigate the molecular mechanism underlying apoptosis of lung adenocarcinoma cell line A549 induced by rapamycin, a type of mammalian target of rapamycin (mTOR) signal pathway inhibitor, combined with hypoxia treatment. Methods:Rapamycin was applied to treat A549 cell line under hypoxia for 16 h. Then the discrepancy of cell apoptosis between treated and untreated control was compared by Hochest 33258 staining. After being treated with rapamycin combined with hypoxia, the mRNA and protein expressions of survivin, a kind of anti-apoptotic molecule, were tested by real-time PCR and Western blotting. Western blotting was employed to test the difference of hypoxia-inducible factor (HIF)-1α expression level before and after rapamycin was added. In additon, chromatin immunoprecipitation(ChIP) assay was further applied to explore the mechanism underlying the effects of rapamycin under hypoxia in inducing apoptosis of A549 cells. Results:Rapamycin induced apoptosis of A549 cell line under hypxia in vitro. In A549 cell line, the expressions of survivin at mRNA (P<0.01) and protein levels, were both decreased by rapamycin combined with hypoxia treatment. Besides, ChIP assay revealed that rapamycin inhibited the expression of survivin at the transcriptional level through decreasing the hypoxia-induced up-regulation of HIF-1α and its binding with the promoter of survivin gene. Conclusion:Rapamycin exerted anti-tumor effects by inhibiting the expression of HIF-1α and decreasing the binding of HIF-1α with the promoter of survivin gene, thereby down-regulating the increase in the expression of apoptosis inhibiting gene survivin.

Objective To observe the behavioral effects of acute administration of dizocilpine (MK-801) in rats,and to determine an appropriate doses of the drug to mimicking memory deficits of schizophrenia.Methods 192 rats were randomly divided into model groups and control group,and given different doses of MK-801 or saline.Locomotor activity,pre-pulse inhibition (PPI) test and novel object recognition test were studied respectively.Results The spontaneous activity increased in a dose-dependent manner after the treatment of MK-801.The total distances of locomotor activity in 0.3 mg/kg group((127.04 ± 32.35) m) exhibited a significant difference compared with control group((35.34 ± 12.81) m,P ＜ 0.05).MK-801 decreased PPI in dose-dependent manner(P ＜ 0.01).MK-801 0.1 mg/kg((103.45 ± 68.04) ％) and 0.3 mg/kg ((41.55 ± 62.93) ％) groups showed significant differences compared to control group ((200.39 ± 30.97) ％) in PPI.The discrimination index of 0.03 mg/kg group((15.78 ± 6.23) ％) and 0.1 mg/kg group((22.42 ± 3.85) ％) were lower than that in control group((39.42 ±3.86)％,P＜0.05).Conclusion It is necessary for select right doses for model different endophenotypes of schizophrenia by MK-801.0.03 mg/kg of MK-801 is a relatively appropriate dosage to cause recognition memory damage.

Objective To investigate the effects of acivicin on HepG 2 cell apoptosis and the effects of acivcin with cis diaminedichloroplatinum (CDDP) on HepG 2 cell apoptosis and on production of intracellular ROS. Methods HepG 2 cells were treated with acivcin, CDDP, and acivicin combined with CDDP. The cytotoxicity was measured by MTT assay. Flow cytometry was used to detect the intracellular ROS, and the rate of apoptosis was determined by TUNEL assay. Results The concentration of acivicin to cause HepG 2 IC 50 was 1.4 mmol/L, and CDDP was 67 ?mol/L. Significant increases in intracellular ROS content were observed in HepG 2 cells 24 h after treatment with acivicin (1.4 mmol/L), CDDP (67 ?mol/L), or acivicin (1 4 mmol/L) plus CDDP (67 ?mol/L), especially with acivicin (1 4 mmol/L) plus CDDP (67 ?mol/L). HepG 2 cell apoptotic rates induced by acivicin (1 4 mmol/L) at 24, 48, and 72 h were (16 3?3 5), (27 9?4 3), (47 2?3 0), respectively, and a significant difference was observed compared to that of control group ( P

Objective To observe the dynamic changes of P 53 , C myc, Bax, Bcl 2, and NF ?B in surviving HepG 2 cells under the continuous stress of cisplatin for the investigation of the potential role in the resistance of HepG 2 cells to cisplatin. Methods The apoptosis of HepG 2 cells and changes of the apoptosis associated factors including P 53 , C myc, Bax, Bcl 2, and NF ?B in the surviving HepG 2 cells were detected by immunohistochemical and RT PCR assays. Results No obvious change of P 53 and C myc was found in the surviving HepG 2 cells under cisplatin stress, but persistent increase of Bcl 2 and decrease of Bax and a continuous increase of Bcl 2/Bax ratio were found. Markedly positive expression of NF ?B mRNA was found at 7 d after the stress of cisplatin. Conclusion The mechanisms of the surviving HepG 2 cells under continuous stress of cisplatin may be associated with the increase in Bcl 2 and decrease in Bax, specially the continuous increase in Bcl 2/Bax ratio, while the positive NF ?B mRNA expression may be associated with the later events after HepG 2 cell survive.