Objective To investigate apoptosis of HepG2 ceils after transfecfion with LIGHT gene and interferon-γ. Methods LIGHT gene and interferon-γ were transfected into HepG2 cells by liposome mediated method. The HepG2 cells were divided into group A (transfected with LIGHT gene or interferon-γ), group B (transfeeted with LIGHT gene and interferon-γ) and group C (non-transfection group). The apoptosis rate of the HepG2 cells and the expression of Bcl-2 and Caspase-8 were detected 12, 24, 48 hours after transfeetion. Results (1) The apoptosis rates of HepG2 cells at hour 12, 24 and 48 after transfeetion were 18.8% ± 3.5%, 25.7%± 2.8% and 36.4% ±3.6% in group A, 23.8% ±2.4%, 31.1% ±2.1% and42.5% ±4.5% in group B, and 8.7% ± 2.1%, 9.3% ± 1.6% and 10.9% ± 1.2% in group C. There was a significant difference in apoptosis rate among the 3 groups (F = 15.69, 53.33, 48.28, P < 0.01). (2) The expression of Bcl-2 in HepG2 cells at hour 12, 24 and 48 after transfection was 16.4% ± 5.0%, 13.4% ± 3.5% and 8.6% ± 2.3% in group A, 14.7%±3.8%, 9.1% ±2.0% and 4.6% ±2.0% in group B, and 25.3% ±6. 3%, 19.8% ±4.4% and 10.1% ±3.8% in group C. There was a significant difference in the expression of Bcl-2 among the 3 groups (F = 6.19, 12.29, 5.81, P <0.05). (3) The expression of Caspase-8 at hour 12, 24 and48 after transfection were 19.3% ±2.4%, 27.2% ±1.9% and 33.7% ±3.0% in group A, 22.7% ±2.2%, 30.9% ±3.1% and 38.2% ±3.2% in group B, and 1.2% ±0.8%, 1.8% ±0.6% and 3.2% ±1.5% in group C. There was a significant difference in the expression of Caspase-8 among the 3 groups (F =71.54, 112. 78, I01.61, P < 0.01). Condusions LIGHT gene can signiticanfly promote cell apoptosis through regulating the expression of Bcl-2 and Caspase-8. Interferon-γ enhanced the effect of LIGHT gene on the apoptosis of HepG2 cells.

Objective To evaluate the efficacy and safety of a new drug,human urinary kallidinogenase,against acute brain infarction.Method A 15-center,randomized,double-blinded and 3:1 placebo-controlled study was carried out.Acute brain infarction within 48 hours of onset in the territory of the middle cerebral artery were indicated as subjects;kallidinogenase or placebo which was dissolved in 50 ml saline,was slowly injected intraveousely within 30 minutes daily for 3 weeks.The European Stroke Scale and Barthel Index were used to evaluate the neurological deficit and the activities of daily living(ADL),followed by a follow-up at the end of the third month.Results 446 patients were enrolled,who completed ITT analysis,including 330 in kallidinogenase group and 116 in placebo group,meanwhile 421 proceeded with PP analysis(311 and 110 respectively).There were no significant differences of the baseline data between the 2 groups.At the end of treatment,the ESS scores increased by 55.1%?33.0% and 44.7%?32.8% respectively in kallidinogenase group(KG)and placebo group(PG,P=0.0022),the difference being significant.PP analysis had similar results.As for ADL,follow-up 90 days after the treatment showed 374 cases followed,280 in KG and 94 in PG;1 died in PG,while none in KG.In KG,the cases whose BI≥50 were significantly more than those in PG(P=0.0228).Adverse events possibly or definitely attributable to the drug were observed in 27 cases(7.74%),mostly were mild,such as palpitation,flush,dizziness, nausea etc,without special management needed.Only 2 died which was confirmed not correlated to kallidinogenase,and another 2 cases of sudden blood pressure drop were observed.The blood pressure drop, quickly restoring soon after the withdrawal of kallidinogenase and use of hemopiesic drugs,was considered to be caused by the combination use of anti-hypertensive drug ACEI and quick infusion speed.Conclusion Kallidinogenase is efficacious for acute brain infarction in improving the neurological deficits,which is safe in clinical use.

Objective To investigate effect of secretin-enhanced magnetic resonance cholangiopancreatography (S-MRCP) on visualization improvement of main and branch pancreatic duct and its diagnostic value of grading chronic pancreatitis (CP) .Methods The 3 .0 T magnetic resonance (MR) imaging data of 88 subjects (23 health volunteers and 65 CP patients) were prospectively analyzed .All the subjects received coronal T2-weighted sequences scan ,thick slab 2-dimensional MRCP sequences scan , routine pancreas plain scan and enhanced scan before and after 0 .1 mL/kg secretin intravenous injection . Sixty-five CP patients were grouped according to Cambridge classification and also according to normal and abnormal of duodenal filling (DF) .The images of main pancreatic duct (MPD) ,branch pancreatic duct (BPD) and DF were compared before and after secretin injection .The result was compared with that of endoscopic retrograde cholangio pancreatography (ERCP) which was the gold standard .One-way analysis of variance test was performed for comparison between healthy control group and CP group .Chi-square test was used for comparison between normal DF group and abnormal DF group .The consistence between the diagnostic results of MRCP ,S-MRCP and the results of ERCP were analyzed by Kappa test .Results Compare with MRCP ,after secretin injection the indexes of MPD ,the sensitivity ,specificity ,positive predictive value and negative predictive value of BPD were all improved ,and the rates of misdiagnosis and missed diagnosis all decreased .Using ERCP as gold standard of CP grade diagnosis ,the consistence of S-MRCP in CP grade diagnosis increased more significantly (Kappa=0 .77 ,P=0 .000 1) compared with that of MRCP (Kappa=0 .55 ,P=0 .000 1) .Among 43 normal DF cases ,the number of MPD dilatation , filling defect and branch duct in pancreatic head was 28 ,23 and 30 ,respectively ;among 22 abnormal DF cases which was 20 ,19 and 21 ,respectively ,and the differences were statistically significant (χ2=5 .01 , 3 .91 ,6 .88 and 4 .26 ,all P<0 .05) .Taking 4 min as the cutoff value of peak time ,the sensitivity and specificity of which in CP diagnosis were 86 .0% and 100 .0% , respectively . Conclusion S-MRCP improves the visualization of MPD and BPD ,and also increases the accuracy of CP grade diagnosis .

AIM: To construct a knock-out vector for ?~(maj) & ?~(min) gene of ?-globin gene and establish embryonic stem cell lines by gene knock-out in ES level for generating gene knock-out animal model. METHODS: Two homologous sequences 1.7 kb and 7.0 kb were inserted into basic plasmic vector pNTK, which contains positive and negative selection system of the neomycin resistance (Neo) and herpes simplex virus thymidine kinase (TK) resistant gene to construct a gene knock-out recombinant plasmic vector nominated ?-pNTK, which was certified by polymerase chain reaction, enzyme digested and DNA sequencing. The ?-pNTK that contains two homologous sequence was linearized with Sal I and introduced into the D3 line of ES cells by electroporation. Positive cloning of ES cells was selected with positive and negative selection system of G418 600 mg/L and 2.5 mol/L Gancyclovir for four weeks. ES cells clone were picked, explanded and certified by polymerase chain reaction, Southern blotting. Examination of the undifferentiated state and pluripotential characteristics of the cell lines were made with the alkaline phosphatase (AKP) staining and embryoid bodies formation test and teratomas formation in nude mice. RESULTS: A knock-out recombinant plasmic vector for ? maj & ? min gene of ?-globin gene, ?-pNTK, was constructed in C129 mouse, which contains 8.7 kb length inserted homologous sequence and deleted most part of the ?~(maj) & ?~(min) gene. 8 strains of positive ES cells cloning were obtained, all of them were testified positive with polymerase chain reaction and Southern blotting. The results of AKP staining, embryoid bodies formation in vitro and teratomas formation in nude mice showed undifferentiated state and pluripotential characteristic of cell line. CONCLUSIONS: A knock-out recombinant plasmic vector of ?~(maj) & ?~(min) of ?-globin gene in C129 mouse was constructed, in which a knock-out ?~(maj) & ?~(min) gene of ?-globin gene ES-D3 cell line was established by electroporation, characterization of these cells show a prospect utility in producing gene knock-out mice in blastocysts stages.

Objective:To study the chemical constituents in Yinqiaosan Powder that have antiviral action on influenza.Method:Isolation of the constituents by different kinds of column chromatography and elucidation of their structures by chemical and spectral methods.Result:Seven flavonoids were obtained,and six of them were identified as linarin，acacetin，hesperidin，isoquiritigenin，isoquiritin and hyperoside respectively.Conclusion:Linarin and acacetin were isolated from Yinqiaosan Powder for the first time.

An extremely thermoacidophilic isolate K4-1 was obtained from an acidic hot spring in Teng- chong Rehai, Yunnan province. Morphology, growth characteristics, utilization of carbon compounds, en- ergy sources and 16S rRNA gene sequence of K4-1 were studied. Cells of K4-1 are irregular cocci with monotrichous flagella. The strain grew aerobically in either a lithotrophic or a heterotrophic mode. Growth on elemental sulfur occurred through oxidation of sulfur. It grew optimally at 75?C and pH 3.5. On the basis of 16S rRNA gene sequence similarity, strain K4-1 was shown to belong to genus Sulfolobus, being related to the type strains of genus Sulfolobus (86.6%~94.3% similarity), and being most closely related to strain Sulfolobus tengchongensis RT8-4 (98.9% similarity). The GenBank accession number of strain K4-1 16S rRNA gene sequence is EU729124.

AIMTo study the binding of sibutramine hydrochloride (SH) and bovine serum albumin (BSA) in physiological condition by spectroscopic method.

METHODSThe quenching mechanism of the fluorescence of bovine serum albumin by sibutramine hydrochloride was studied with the fluorescence and the absorption spectroscopy. The binding constants K and the number of binding sites were determined at different temperatures according to Scatchard equation and the main binding force was discussed by thermodynamic equations. The effect of the drug on bovine serum albumin conformation was also studied by using synchronous fluorescence spectroscopy.

RESULTSThe quenching mechanism of sibutramine hydrochloride to bovine serum albumin was static quenching. The binding constants K at 8 degrees C, 25 degrees C, 37 degrees C were 1.21 x 10(5), 8.31 x 10(4), 6.97 x 10(4) L x mol(-1) with one binding site, respectively. The thermodynamic parameters of the reaction were deltaH = -9.70 kJ x mol(-1), deltaS = 56.41 J x mol(-1) x K(-1).

CONCLUSIONThe binding force is electrostatic interaction. Sibutramine hydrochloride can be deposited and transported by serum protein in vivo. Sibutramine hydrochloride has nearly no effect on the serum protein conformation.

Animals ; Binding Sites ; Cattle ; Cyclobutanes ; metabolism ; Protein Binding ; Serum Albumin, Bovine ; metabolism ; Spectrometry, Fluorescence ; methods ; Spectrophotometry, Ultraviolet ; methods

Objective To evaluate the efficacy and safety of oxycodone hydrochloride for postoperative analgesia in patients who received spin surgery.Methods Sixty -nine patients treated with spin surgery were included in this study and randomly divided into treatment group ( n=34 ) and control group ( n=35).Half an hour before the end of operation ,the patients of the two group were given oxycodone hydrochloride and morphine 0.07 mg · kg-1 , respectively.After extubation , patient -controlled analgesia pump was given with the background dosage of 0.5 mg · h -1 and one time push 1 mg of oxycodone hydrochloride or morphine.The mean arterial pressure ( MAP) , and heart rate were com-pared in the time points of entering the operation room ( T0 ) ,tracheal in-tubation ( T1 ) , 5 min after tracheal intubation ( T2 ) , and 15 min after tra-cheal intubation(T3) between the two groups.The recovery time, extu-bation time ,visual analogue scale ( VAS) ,observer assessment of sedation ( OAA/S ) score and safety were compared between the two groups . Results The MAP and heart rate were not statistically different between the two groups and among the time points ( P>0.05 ).The recovery time , extubation time , VAS, OAA/S score in treatment group were significant lower than those of control group( P<0.05 ).The adverse drug reaction rates were 17.14%and 8.82% in the control group and treatment group respectively with statistical difference ( P<0.05 ).Conclusion Compared with morphine , oxycodone hydrochloride is more effective and safe for postoperative analgesia in patients who received spin surgery.

OBJECTIVETo study the chemical constituents with antiviral action from Yinqiaosan on influenza virus.

METHODConstituents were isolated by different kinds of column chromatography and their structures were elucidated with chemical and spectral methods.

RESULTEleven chemical constituents were isolated and elucidated as arctiin, phillyrin, forsythiaside, liquiritigenin, liquiritin, genistein, formononetin, daidzein, glycitrin, 3,3',4-tri-omethlellagic acid and chlorogenic acid.

CONCLUSIONGenistein, daidzein, glycitrin and 3,3',4-tri-omethlellagic acid were isolated from Yinqiaosan for the first time.

Antiviral Agents ; chemistry ; isolation & purification ; Arctium ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Forsythia ; chemistry ; Furans ; chemistry ; isolation & purification ; Genistein ; chemistry ; isolation & purification ; pharmacology ; Glucosides ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Isoflavones ; chemistry ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry

OBJECTIVETo look for and determine the distinctive compound in Pinellia ternata.

METHODWith TLC, HPLC, the distinctive compound was found and obtained by using the method of HPLC preparation. Its structure was elucidated by spectral analysis and physico-chemical properties.

RESULTThe compound was identified as inosine.

CONCLUSIONInosine is the distinctive compound in Rhizome of P. ternata, and it was isolated from Banxia for the first time.

Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Inosine ; chemistry ; isolation & purification ; Molecular Structure ; Pinellia ; chemistry ; Plants, Medicinal ; chemistry ; Rhizome ; chemistry