Objective To construct an X-gene defect and IFN gene contained HBV expression plasmid and explore its expression effect in HepG2 cell line. Methods By employing pBR322-B-HBV plasmid which contained type adrⅠHBV DNA, we destroyed HBV X-gene by inserting mutation and connected the defect HBV with IFN sequence. After recombination, it was introduced into a mammalian plasmid and was constructed as the objective plasmid. At the same time, two control groups were constructed. One was pcDNA3-ES-HBV2, which contained the full length of HBV DNA. The other was pcDNA3-KN-F1F2, which contained the X-gene defect HBV DNA. Then we transfected HepG2 cells with all the plasmid. After the cells were screened by G418, the X-gene defect and IFN gene contained HBV(KN-F1F2-IFN) and IFN protein from culture medium were detected by fluorescence- quantitative-PCR and ELISA respectively. Results pcDNA3-KN-F1F2-IFN, as the objective plasmid, was constructed successfully; contrary with the control virus, the amounts of both HBV and HBsAg were lower; both IFN mRNA and IFN protein were detected.Conclusions An X-gene defect and IFN gene contained HBV expression plasmid was constructed successfully and it can be expressed in HepG2 cell line.

OBJECTIVE:To investigate the effect of STAT5 pathway inhibitor pimozide on the expressions of nitric oxide (NO)and nitric oxide synthetase(iNOS)in the model of mouse macrophage RAW264.7 inflammation induced by lipopolysaccha-ride (LPS). METHODS:RAW264.7 cells in logarithmic growth phase were divided into blank control group,drug control group (10μmol/L pimozide),model group(1μg/ml LPS)and the pimozide groups of low,middle and high doses(2.5,5 and 10μmol/L), where the corresponding cells were given pimozide 30 min before the administration of LPS,and then were cultured for 24 h. Griess method was used to determine the content of NO in the supernate of cell culture solutions of all groups,real-time quantita-tive polymerase chain reaction(RT-PCR)to determine iNOS mRNA expression,and Western blot method to determine the protein expression of iNOS and phosphorylated STAT5(p-STAT5). RESULTS:The content of NO,iNOS mRNA and protein expressions and the content of p-STAT5/STAT5 in the cells in the model group were higher than those in the blank control group,with statisti-cally difference (P<0.01). Compared to the model group,the pimozide groups of middle and high doses had lower content of NO,iNOS mRNA and protein expressions and the content of p-STAT5/STAT5 in the cells,with statistically difference(P<0.01 or P<0.05). CONCLUSIONS:STAT5 pathway inhibitor pimozide can inhibit the release of NO by inhibiting iNOS mRNA and pro-tein expressions in cells.

Objective To evaluate the efficacy of calcium(Ca)and magnesium(Mg)infusions in pre- vention of oxaliplatin-induced acute neurotoxicity.Methods Fourty-two patients with advanced colorectal carcinoma were eligible for the study:21 were assigned to the Ca/Mg ann and 21 to the control arm.The Ca/ Mg arm and the control arm were comparable for patients'characteristics.Chemotherapy regimen were al- most the same in both arms.Chemotherapy regimen consisted of oxaliplatin 130 mg/m~2 on day 1,given as a 3-hour infusion in 500 ml of 5% glucose,concurrent with rahitrexed 3 mg/m~2 as a 15-minute intravenous (Ⅳ)infusion or calcium folinate(CF)200 mg?m~(-2)?d~(-1),days 1～5,5-Fluorouracil(5-Fu)500 mg?m~(-2)?d~(-1),days 1～5.Therapy was repeated every 3 weeks.The treatment consisted of Ca gluconate and Mg sulfate,1 g each, delivered i.v.over 15 min just before the oxaliplatin infusion and repeated at the same dose after the comple- tion of the oxaliplatin infusion.A specific neurotoxicity scale was used for oxaliplatin-related neurotoxicity. Results Ten patients(47.62%)had acute neurotoxicity in the Ca/Mg arm compared with 19 patients (90.48%)in the control arm(P0.05).Conclusion Ca/Mg infusions seem to reduce incidence and intensity of oxaliplatin-induced acute neurotoxicity,and they do not reduce the clinical activity of oxaliplatin,but dosage and administration schedule could be optimized.

Slowly growing mycobacteria (SGM ) are distributed in the environment ,for example in soil and dirty water . SGM can cause human infections ,especially lung diseases .In this article ,first and second line antituberculous agents were ex‐amined in order to identify the optimum drugs for the treatment of SGM disorders .The fewest SGM in our study (4/34) were susceptible to isoniazid .Rifampicin (13/34) and ethambutol (14/34) were effective against similar numbers of strains .Ofloxa‐cin (23/34) ,kanamycin (26/34 ) , tobramycin (26/34 ) and streptomycin (27/34 ) were active against most of the tested strains .Ciprofloxacin (31/34) ,levofloxacin (31/34) ,amikacin (33/34) and capreomycin (33/34) showed an excellent range of activity .Moxifloxacin (34/34) showed the widest range of activity against the SGM species .Among the tested SGM spe‐cies ,M .simiae and M .af ricanum were resistant to the highest number of drugs .M .szulgai and M .duvalii were susceptible to all the first and second line antituberculous agents tested .Overall ,the second‐line antituberculous agents were good candi‐dates for the treatment of infection by SGM species and can be widely used in the therapy of SGM diseases .

Objective To investigate natural killer(NK) and NKT cells in acute pancreatitis(AP).Methods Changes of NK and NKT cells in peripheral blood of 86 AP cases were detected using muhiparameter flow cytometry.Results Compared with control group,the NKT cells decreased in AP patients (t =5.23,P =0.00),but NK cells didn't (t =-1.15,P =0.25).NKT cells in severe SAP and mnoderate MAP were lower than that in the control group (t =-3.92,P =0.00;t =4.84,P =0.00).There was no statistically significant difference of NK cells between MAP and the controls (t =-0.54,P =0.59),but NK cells in SAP group was obviously higher than that in control group (t =3.12,P =0.00).After one week treatment,NK cells significantly decreased (t =8.43,P =0.00).NKT cells were higher than control group (t =-4.44,P =0.00).Dynamic monitoring in AP patients found continuous declination in NK cells,and NKT cells experienced an increase before a falling.Conclusion Monitoring of NK and NKT cells can be used as an important index for the severity and response to treatment in acute pancreatitis.

The current study to separate and identify constituents from Millettia nitida var. hirsutissima. The compounds from Millettia nitida var. hirsutissima were isolated by means of various chromatographic techniques such as column chromatography over ODS and Sephadex LH-20, preparative HPLC, and the structures of these isolated compounds were identified through spectroscopic analyses. Nine isoflavonoids and two flavans were isolated and identified as 5-O-methy genistein (1), 7-hydroxy-3',4'-dimethoxyisoflavone (2), ononin (3), catechin (4), formononetin (5), genistein (6), calycosin (7), (-)-gallocatechin (8), sissotrin (9), wistin (10), daidzin (11). Compounds 1, 2, 9 are obtained from the genus Millettia for the first time,and compounds 4, 8 are isolated from this plant for the first time.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Flavones ; chemistry ; isolation & purification ; Isoflavones ; chemistry ; isolation & purification ; Mass Spectrometry ; Millettia ; chemistry ; Molecular Structure

Background Retinopathy of prematurity is mainly due to retinal neovascularization.Objective This laboratory work was to evaluate the efficacy of different dosage of avastin for inhibiting retinal neovascularization.Methods Ninety 7-day-old clean C57BL/J6 mice were randomized into six groups as follows:air control group,hyperxia control group,hyperxia BSS group and avastin groups.C57BL/J6 mice in air control group were raised in regular air environments.The fifty mice were fed under the environment with 75％ ±2％ oxygen for 5 days to establish the retinal neovascularization models.The 1.25,2.50 and 5.00 g/L avastin (0.5 μl) were injected inteavtreally in forty-five mice models as low,moderate and high dosage avastin groups respectively,and 0.5 μl BSS was used at the same way in fifteen models as hyperxia BSS group.The mice were sacrificed in the 17-day-old age using excessive anesthesia method and the retina sections were prepared for the calculation of the numbers of vascular endothelial cell nuclei broken retinal inner membrane after hemotoxylin and eosin staining.The expression of CD34 in the retina was detected by immunochemistry.The morphology and distribution of retinal neovascular vessel in various groups were observed using retinal flat.The use of the animals followed the Regulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results The numbers of cell nuclei broken the inner limiting membrane was significant increased in the hyperxia group compared with the air control group( P＜0.01 ),and those in difference doses of avastin were considerably reduced in comparison with hyperxia BSS group (P＜0.01) and hyperxia group (P＜0.01 ).The decrease of numbers of cell nuclei broken the inner limiting membrane was obvious in low dose of high dose of avastin compared with low dose of avastin (P＜0.05 ).CD34 was positively expressed in retina internal membrane of hyperxia group.Retinal flat revealed the regular distribution and normal structure of retinal vessels in air control group and avastin groups.However,retinal and vitreous cavity neovascularization,leakage and enlarged non-perfusion regions in the perimeter of the retina were seen in hyperxia group and hyperxia BSS group. Conclusions Intravitreal injection of avastin can arrest retinal angiogenesis in oxygen-induced retinal neovascularization models in a dose-dependent manner.

The uhrastructure and atrial natriuretic peptide(ANP)mRNA in myocardium of streptozotocin-induced diabetic rats were studied along with the effect of rosiglitazone.It was found that the myocardial ultrastructure was demaged in diabetic rats.Rosiglitazone may ameliorate the lesion in myocardium,and lower the ANP mRNA in diabetic rats(1.14?0.05 at 6 weeks and 1.12?0.09 at 10 weeks vs 0.97?0.14,both P

Objective To explore the influence of G4 cyberknife treatment of large hepatocellular car-cinoma on liver function,and to evaluate its treatment safety.Methods Sixty-three large liver cancer patients treated with routine G4 cyberknife treatment were retrospectively analyzed,and then statistical analysis of the difference in liver function before and after treatment was conducted.Results After G4 cyberknife treatment of 1 2 months,the levels of ALT,ALP,TBIL,PA were respectively 23.00 U /L,1 1 1 .00 U /L,1 3.70 μmol/L, (81 .87 ±1 3.94)%.Compared with the levels before treatment [28.00 U /L,32.00 U /L,1 1 .30 μmol/L, (86.07 ±1 4.07)%],there were no signi-ficant differences found (Z =-1 .677,P =0.094;Z =-0.504, P =0.61 4;Z =-1 .945,P =0.053;t =1 .271 ,P =0.21 3).The level of ambumin was (34.84 ±4.75)g/L at 1 2 months after treatment,which decreased and the difference compared with the level before treatment [(37.45 ±4.1 4)g/L]was significant (t =3.357,P =0.002).The Child-Pugh grade was 5.80 ±1 .1 7 respectively at the time points of 1 2 months after treatment,and no significant difference was found compared with the Child-Pugh grade before treatment (5.48 ±0.81 ,t =-1 .668,P =0.1 06).Conclusion G4 cyberknife treatment does not cause liver injury.It is safe and reliable in large liver cancer treatment.So,it is worth widely clinical popularizing.

Background and purpose:TrkB,a neurotrophic tyrosine kinase receptor,serves as a potent and specific suppressor of anoikis of non-malignant epithelial cells,and induces high invasion capacity in these cells.TrkB over-expression has been associated with chemotherapy resistance and poor survival in neuroblastoma and some other highly aggressive cancers.However,the relationship of the expression of TrkB to anoikis resistance in ovarian cancer cells has rarely been reported in literature.This study investigated the expression and significance of anoikis- suppressor TrkB in OVCAR-3 ovarian cancer cells.Methods:The expression of TrkB and its ligand BDNF was evaluated in OVCAR-3 ovarian cancer cells under different culture conditions by RT-PCR and Western blot.Results: TrkB mRNA was overexpressed in multicellular spheroids(cell-spheroids,anchorage-independent culture,AIC)as compared to that in OVCAR-3 cells(adhesive-cells,adhesive culture,AC),(35.3?0.7)% versus(23.5?0.5)%;but BDNF mRNA expression was the opposite to the above situation,(41.4?0.6)% versus(32.24?0.7)%(P