Objective:To study the changes of PTEN gene expression in radiation-induced bystander cells.Methods:The irradiated cells and non-irradiated cells were co-cultured in a ratio of 11.The culture medium of irradiated cells was used to culture non-irradiated cells to create two bystander cell models. The expression of PTEN mRNA and protein in the bystander cells,irradiated cells,non-irradiated cells was compared.Results:The protein expression of PTEN in the irradiated cells was negatively associated with the irradiation dose.The down-regulation degree of PTEN protein was related to the different treatments of cells.The most prominent down-regulation was found in the bystander cells cultured with medium of irradiated cells.The down-regulation of PTEN protein in the two kinds of bystander cells was more severe than that in the irradiated cells(P

Objective To examine the expression of ATP-binding cassette transporter A1(ABCA1)in eukaryotie cells and the effect of arsenic resistance after the transfection of eukaryotic expression vector containing ABCA1 gene.Methods HeLa cells were transfected with the recombinant plasmid by lipofectaonmine 2000 (recombinant plasmid group),empty plasmid and untransfected HeLa cell as the control group.The level of the mRNA was examined by real-time PCR,and the expression of ABCA1 protein wag examined by Western blot,the change of cell survival rate was examined by methyl thiazolyl tetrazolium(MTT)after exposure in a series of arsenic [0(contro1),4,8,16,32,64,128 μmol/L]for 48 hours.Results Expression level of ABCA1 mRNA in recombinant plasmid,empty plasmid and untransfeeted groups was(2.09±0.08)×10-4,(0.09±0.02)×10-4,(0.08±0.02)×10-4,there was a significant difference between the groups(F=1499.23,P<0.01).The level of ABCA1 mRNA in recombinant plasmid group was higher than empty plasmid and untransfected group(all P<0.01).Western blot showed that specific protein straps existed at 254×103 in all the three groups,with a similar size to the ABCA1 protein.The amount of the recombinant plasmid group was higher than the other two groups.MTT shows that arsenic concentration at 4,8,16,32,64,128 μmol/L,the survival rates of recombinant plasmid group was(94.8±0.9)%,(86.5 ± 2.6)%, (77.8 ± 2.0)%, (56.0 ± 2.0)%, (23.8 ± 1.7)%, (18.6 ± 0.6)%, higher than that of empty plasmid group[ (85.3 ± 1.1)%, (78.7 ± 0.6)%, (67.8 ± 2.4)%, (43.2 ± 1.5)%, (14.5 ± 1.3)%, (8.0 ± 0.4)%], the difference of survival rate had a statistical signifieance(t = 18.985,6.689,5.922,9.504,9.481,32.634, all P < 0.01). Conclusions ABCA1 protein is over expressed in HeLa cells after transfect ABCA1 gene. ABCA1 protein increases resistance of arsenic in HeLa cells.

To establish an HPLC-MS/MS method for the analysis of quercetin, kaempferid and isorhamnetin in rats plasma and study its pharmamacokinetics after an intragastrical administration of Hippophae rhamnoides extracts. Five healthy male Sprague-Dawley (SD) rats were given single doses of H. rhamnoides extracts (quercetin 26.35 mg x kg(-1), kaempferid 4.040 mg x kg(-1), isorhamnetin 31.37 mg x kg(-1)), and then their orbital sinus blood samples were collected at different time points. The drug plasma concentration of the three flavonoids was determined by HPLC-MS/MS method. After that, the main pharmacokinetics parameters were calculated by using Kinetica 5. 0. 11 software. The methodological test showed that the linear concentration ranges of quercetin, kaempferid and isorhamnetin were 7.500-600.0 μg x L(-1) (R2 = 0.998 5), 1.000-80.00 μg x L(-1) (R2 = 0.998 5 ) and 10.00-800.0 μg x L(-1) (R2 = 0.998 0), respectively. The inner and inter-days precisions were both less than 14.0%. The plasma samples showed a good stability and consistency with the requirement of biological sample analysis after the samples were frozen once and placed at - 20 degrees C for 15 d and room temperature for 6 h and the treated analytes were placed at -20 degrees C for 24 h. For quercetin, the pharmacokinetic parameter t(½β), AUC(0-∞), MRT(0.∞), C.(max) and T(max) were (113.3 ± 19.37) min, (12 542.14 ± 3 504.05) μg x h x L(-1), (119.6 ± 13.29) h, (164.6 ± 27.33) μg x L(-1) and (5.199 ± 0.840 3) h, respectively. For kaempferid, the pharmacokinetic parameters t(½β), AUC(0-t), MRT(0-∞), C(max) and T(max) were (79.85 ± 17.15) min, (934.51 ± 94.59) μg x h x L(-1), (81.50 ± 13.75) h, (80.15 ± 14.24) μg x L(-1) and (3.827 ± 0.902 7) h, respectively. For isorhamnetin, the pharmacokinetic parameters t1,2,, AUC(0-t), MRT(0-∞), C(max) and T(max) were (118.3 ± 20.73) min, (26 067.77 ± 4 124.60) μg x h x L(-1), (129.0 ± 16.30) h, (269.6 ± 29.32) μg x L(-1) and (6.513 ± 1.450) h, respectively. The HPLC-MS/MS analysis method established in this study was proved to be sensitive and accurate and could be applied in the pharmacokinetic study of quercetin, kaempferid and isorhamnetin in rat plasma.
Animals ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; pharmacokinetics ; Hippophae ; chemistry ; Kaempferols ; blood ; pharmacokinetics ; Male ; Quercetin ; analogs & derivatives ; blood ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; methods

OBJECTIVE To strictly manage the whole process for cleaning the laminar air flow(LAF) operating center to achieve the modernized hospital environment′s standard requests.METHODS Right before the use of the LAF operating center,scientific management was conducted strictly according the regulations and standards issued by state Ministry of Health.RESULTS Fulfilling the standards of the process for cleaning operating center was all for the goal of improving the efficiency of management.CONCLUSIONS A management is made efficient from checking all the things before use,monitoring them,and training people so that they can efficiently carry out their tasks.The purpose of the scientific management is to reach the expected quality.

OBJECTIVETo investigate the association of Coxsackie B virus (CBV) with habitual abortion.

METHODSCBV IgM antibody, viral RNA and virions were detected in 86 women with habitual abortion and 40 with induced abortion by means of enzyme-linked immunosorbent assay (ELISA), RT-PCR and virus isolation, respectively.

RESULTSThe positivity rate of CBV IgM were 87.2% and 35% in the two groups, respectively, and the detection rate of the viral RNA was 53.5% and 17.5% in blood lymphocytes, and 59.3% and 17.5% in the placentas. The virions were found in the placentas in 41.9% and 6.9% of the women, respectively. The positivity rates of CBV IgM, viral DNA and virions showed significant difference between the two groups (P<0. 01).

CONCLUSIONCBV might be one of the causes responsible for habitual abortion.

Abortion, Habitual ; blood ; etiology ; Adult ; Antibodies, Viral ; blood ; Coxsackievirus Infections ; complications ; virology ; Enterovirus B, Human ; immunology ; isolation & purification ; Female ; Humans ; Immunoglobulin M ; blood ; Lymphocytes ; ultrastructure ; virology ; Microscopy, Electron ; Pregnancy ; Pregnancy Complications, Infectious ; blood ; virology ; RNA, Viral ; blood

OBJECTIVETo determine the physiological role of D-bifunctional protein (DBP) in bile acid biosynthesis through investigating the effect of increasing activity of DBP on bile acid biosynthesis.

METHODSTwenty male Wistar rats were divided into two groups: diethylhexyl phthalate (DEHP) group (n = 10) and control group (n = 10). Serum triglyceride, total cholesterol, hepatic DBP activity, and fecal bile acids were assayed. The mRNA levels of hepatic peroxisome proliferator-activated receptor alpha (PPARalpha), DBP, and cholesterol 7alpha-hydroxylase (CYP7A1) were detected by RT-PCR.

RESULTSCompared with control group, serum triglyceride level was decreased significantly and PPARalphamRNA level was increased significantly in DEHP group (P < 0.01). Together with a sharp induction of DBP mRNA expression and DBP activity in DEHP group (P < 0.01), the levels of CYP7A1 mRNA and fecal bile acids were significantly increased by 1.9 times and 1.6 times respectively compared to control group (P < 0.01). There was a significantly positive correlation between DBP mRNA level or DBP activity and CYP7A1 mRNA level (r = 0.89, P < 0.01; r = 0.95, P < 0.01).

CONCLUSIONThe up-regulation of DBP mRNA and activity in liver can result in the increase in CYP7A1 mRNA expression and bile acid biosynthesis, suggesting that DBP may be involved in bile acid biosynthesis together with CYP7A1.

17-Hydroxysteroid Dehydrogenases ; metabolism ; Animals ; Bile Acids and Salts ; biosynthesis ; Cholesterol 7-alpha-Hydroxylase ; analysis ; Enoyl-CoA Hydratase ; metabolism ; Liver ; metabolism ; Male ; Multienzyme Complexes ; metabolism ; PPAR alpha ; analysis ; Peroxisomal Multifunctional Protein-2 ; RNA, Messenger ; analysis ; Random Allocation ; Rats ; Rats, Wistar

Objective To investigate the expression of FHIT gene in the 60Co gamma-ray irradiated human lymphocytoblast(AHH-1) cell and the bystander effect cell,and to explore the function of FHIT gene in the bystander effect of ionizing radiation.Method Preparation of bystander effect cell model:after irradiated with different dose of 60Co gamma-ray(0,2,5 Gy),the directly irradiated AHH-1 ceils were collected immediately by centfifugation and co-cultivated with non-irradiated cells in Transwell.forming the bystander effect group P1.In addition,some culture media supernatant of direcfly irradiated cells were transfefred to the non- irradiated cells culture medium,forming the group P2.Then cells were collected at 0,6,12,and 24 h after irradiation and the total RNA and protein were extracted.RT-PcR and Western blot were performed to determine the FHIT mRNA and protein level.respectively.Flow cytometry assay and cell counting were conducted to detect the alteration of cell cycle and cell proliferation,respectively at 0,24 h after irradiation.Results The mRNA level of FHIT gene among control cells,directly irradiated cells and bystander cells showed no obvious difference. while the FHIT protein level of the directly irradiated ceils and bystander cells was siguificandy down-regulated compared with the control cells(F=102.45,P<0.001).Moreover,the directly irradiated cells and bystander cells showed significant G2 phase arrest and obviously inhibited the proliferation ability.Conclusions 2 and 5 Gy of 60Co γ-ray irradiated AHH-1 cells can result in down regulation of the FHIT protein expression,which suggests that FHIT gene is involved in the process of bvstander effect induced by irradiation.

Objective: To observe the influence of moxibustion sertma of mice on the expression of Fas, bcl-2 mRNA and protein of EL-4 lymphoma cells in vitro. Method: The EL-4 lymphoma cells were cultivated for 24 h in the moxibustion serum of mice. The expression of Fas and bcl-2 mRNA of EL-4 lymphoma cells were detected by in-situ hybridization method, and the protein expression of Fas and bcl-2 were observed by the immuocytochemistry method. Results: The expression of bcl-2 mRNA and protein decreased, and the expression of Fas rnRNA and protein increased significantly in EL-4 cells, which were cultivated in the moxibustion serum compared those cultivated in normal mice serum (P<0.05). Conclusion: Moxibustion serum could down-regulate the bcl-2 mRNA and protein and up-regulate the Fas mRNA and protein of EL-4 cells.

Objective To detect the levels of cytokine IL-2,IL-12,IFN-?and IL-4 secreted by peripheral CD8~+ T lymphocytes in patients with condyloma acuminatum (CA).Methods Flow cytometry was employed to study the expression of cytokines IL-2,-12,INF-?and IL-4 in CD8~+ T lymphocytes in the peripheral blood of 60 patients with CA and 20 healthy controls.Results The percentage of CD8~+ T lym- phocytes producing IL-2,IL-12 and IFN-?were significantly lower in CA patients than that in healthy con- trols (P

Neuritin is a new neurotrophic factor found recently. In order to identify the function of Neuritin clearly, the coding sequence of human neuritin was amplified by PCR from neuritin cDNA , this fragment digested by NocI and NotI was inserted into pET32a by T4 ligase and transformed into E. coli BL21 then the recombinant plasmid named pET32a-neuritin was constructed successfully . Neuritin was expressed distinctly after inducing by EPTG. The product was identified as neuritin by SDS-PAGE and Western blot analysis . The expression production was purified on Ni2+-NTA column.