OBJECTIVE: To investigate the blocking effect of bergapten on cell cycle in nasopharyngeal carcinoma (NPC) cells. METHODS: The inhibition was tested by CCK-8 assay after the NPC cells (CNE-2 and HONE-1) being exposed to bergapten for 48 h. The bergapten concentrations were as follows: 1, 10 and 100 μmol·L-1. The cell cycle was assayed by PI staining and following flow cytometry analysis, and the CDK4, CDK6, CDK2, Cyclin D1, Cyclin E2, β-catenin, GSK-30 and p-GSK-3p (S9) protein levels were detected by Western blotting. RESULTS: Bergapten was able to significantly inhibit CNE-2 and HONE-1 NPC cancer cell proliferations, and the effect was dose-(CNE-2: r=0.926, P<0.05; HONE-1: r=0.959, P<0.05) and time-dependent (CNE-2: r=0.991, P<0.05; HONE-1: r=0.963, P<0.05). Drug treatment induced a block in the G0/G1 phase and decreased protein levels of CDK4, CDK6, CDK2, Cyclin D1, Cyclin E2 and β-catenin, but the protein levels of GSK-3β and the phosphorylation of p-GSK-3β (S9) didn't change. CONCLUSION: Bergapten can block cells cycle and inhibit NPC cells' proliferations, and those effects might be related to suppressing of cells cycle-related protein and interfering with the wnt/p-catenin signal pathway.

To construct the Bac-to-Bac expression system of Bombyx mori nucleopolyhedrovirus (BmNPV), a transfer vector was constructed which contained an Escherichia coli (E. coli) mini-F replicon and a lacZ: attTN7: lacZ cassette within the upstream and downstream regions of the BmNPV polyhedrin gene. B. mori larvae were cotransfected with wild-type BmNPV genomic DNA and the transfer vector through subcutaneous injection to generate recombinant viruses by homologous recombination in vivo. The genomic DNA of budded viruses extracted from the hemolymph of the transfected larvae was used to transform E. coli DH10B. Recombinant bacmids were screened by kanamycin resistance, PCR and restriction enzyme (REN) digestion. One of the bacmid colonies, BmBacJS13, which had similar REN profiles to that of wild-type BmNPV, was selected for further research. To investigate the infectivity of BmBacJS13, the polyhedrin gene was introduced into the bacmid and the resultant recombinant (BmBacJS13-ph) was transfected to BmN cells. The budded viruses were collected from the supernatant of the transfected cells and used for infecting BmN cells. Growth curve analysis indicated that BmBacJS13-ph had a similar growth curve to that of wild-type BmNPV. Bio-assays indicated that BmBacJS13-ph was also infectious to B. mori larvae.

Objective To explore the effect of recovery sleep on the executive function after 36 h of total sleep deprivation by event related potential technology.Methods Thirteen healthy male college students participated in two trials. At the first trial normal sleep as control was investigated. At the second trial participants experienced 36 h of sleep deprivation and then accepted 8 h recovery sleep. In each trial six Go/Nogo tests were employed to test the executive control function and the ERP data were recorded. Results There was no statistical difference in behavior and ERP results at each time point as the subjects had normal sleep. After 36 h of sleep deprivation, the behavior results were statistically significant when compared to the baseline. The amplitude and latency of Nogo-N2, Nogo-P3 on Fz electrode, the amplitude and latency of Nogo-P3 on Cz electrode showed statistical significance when compared to the baseline. After 8 h recovery sleep, the average correct reaction time and the Go correct reaction rate had statistical significance compared to 36 h value. The amplitude of Nogo-N2 and Nogo-P3 had no statistical significance compared to the baseline.However,it was of statistical significance[(-6.80 3.95)vs(-3.37 2.63)μV,(10.63±6.62)vs(5.63±5.45)μV,(9.49±7.37)vs(6.08±6.56)μV] compared to 36 h value. The latency of the recovery value of Nogo-N2 and Nogo-P3 was statistically significant[(254.14±15.55)vs(243.08±13.97)ms(382.14±41.07)vs(349.17±30.36)ms,(369.86±26.48)vs(347.48±29.24)ms]compared to the baseline.Conclusion As the time of sleep deprivation is prolonged, the executive function is impaired and the executive function is not completely recovered after 8 h recovery sleep.

Objective To observe the clinical curative effect of Annaohuoxue capsules, a prescription of promoting blood circulation and removing blood stasis, on patients with hypertensive cerebral hemorrhage and explore its safety. Methods Ninety-three patients with hypertensive cerebral hemorrhage,admitted to our hospital from June 2010 to November 201 i,were chosen and randomly divided into Arnaohuoxue capsule treatment group (group A,n=31),Tiandantongluo capsule treatment group (group B,n=32) and placebo control group (group C,n=30); After 7 d of onset,treatments were given,respectively,for 21 d.Hemorrheology,scores of neurological impairment,Glasgow coma scale (GCS) scores were recorded and compared in these groups before and after treatment. Results The plasma viscosity,and aggregation index of R BCs in patients from group A were significantly lower than those in patients from group B (P＜0.05); statistically significant difference on the clinical efficacy was noted between each 2 groups (P＜0.05):the outcome of group A was the best,that of group B was better,and that of group C was the worst.The GCS scores in patients from group A were significantly higher than those in patients from group B and group C (P＜0.05) Conclusion Annaohuoxue capsules can improve the hemorrheology indices and neurological impairment,and decrease the GCS scores in varying degrees,and the treatment efficiency of Annaohuoxue capsules is much better than placebo.

AIMTo prepare of isopropyl myristate (IPM) molecular gels and investigate of its transdermal capability.

METHODSMicrostructure of IPM gels was studied by scanning electron microscope (SEM) and optical microscope (OM). The rheology and thixotropy of IPM gels were investigated by viscosity. Triptolide was used as model drug to investigate its transdermal capability.

RESULTSThe microstructure of IPM gels was a three-dimension network formed by the aggregation of Span 60 in IPM, which was rod-like tubular aggregate. It has good rheology and thixotropy. There was a good linear correlation between the accumulative permeated amount per unit area and the time for triptolide-loaded IPM gels. The permeation process agreed with zero order pharmacokinetics. The average permeability through rat skin for triptolide was 19.26 ng x cm(-2) x h(-1), which was 2.92 times of triptolide unguents obtained commercially available.

CONCLUSIONIsopropyl myristate molercular gel can be formed by span 60 assemblies. Transdermal capability drug-loaded IPM gels was better than that of triptolide unguents.

Administration, Cutaneous ; Animals ; Anti-Inflammatory Agents, Non-Steroidal ; administration & dosage ; isolation & purification ; pharmacokinetics ; Diterpenes ; administration & dosage ; isolation & purification ; pharmacokinetics ; Drug Carriers ; Epoxy Compounds ; Male ; Mice ; Microscopy, Electron ; Myristates ; chemistry ; pharmacology ; Phenanthrenes ; administration & dosage ; isolation & purification ; pharmacokinetics ; Plants, Medicinal ; chemistry ; Rheology ; Skin Absorption ; drug effects ; Tripterygium ; chemistry ; Viscosity

OBJECTIVETo screen enhancer-like sequences from vaccinia virus genome, to construct an expression vector harboring prokaryotic enhancer-like sequence and study the effect of interferon gene expression.

METHODSEnhancer-like element from vaccinia virus genome was obtained by using the chloramphenicol acetyl-transferase cat gene as reporter gene. An expression vector harboring prokaryotic enhancer-like sequence VV1 from vaccinia virus was constructed. Interferon was expressed and assayed.

RESULTSEighteen enhancing sequences were found. From them two enhancer-like sequences with distance and orientation independence property were screened and named VV1 and VV16 respectively. Quantification test showed that the direct and reverse orientation of VV1 could increase the activity of beta-galactosidase with 10.9 and 3.8 times and those of VV16 could increase by 9.0 times and 4.1 times respectively. The enhancing activity of the element was on transcription level. An expression vector harboring prokaryotic enhancer-like sequence VV1 was constructed. Using this vector the antiviral activity of interferon alpha-2b was increased by 2.6 times in comparison with the original expression plasmid.

CONCLUSIONTwo enhancer-like sequences were screened from vaccinia virus genome. Interferon gene was highly expressed by using an expression vector harboring enhancer-like sequences.

Enhancer Elements, Genetic ; Genetic Vectors ; genetics ; Interferon-alpha ; biosynthesis ; genetics ; pharmacology ; Plasmids ; Recombinant Proteins ; Vaccinia virus ; genetics

OBJECTIVETO establish a RP-HPLC method for the determination of calycosin-7-O-beta-D-glucopyranoside in Radix Astragali, and to analyse the calycosin-7-O-beta-D-glucopyranoside content of ten samples of Radix Astragali, collected from different regions.

METHODA Polaris C18(250 mm x 4.6 mm, 5 microns) column was used and a mixture of methanol-water (30:70) was used as the mobile phase at a flow rate of 1.0 mL.min-1. The column temperature was 25 degrees C and the UV detection wavelength was 254 nm.

RESULTThe calibration curve was in good linearity over the range of 0.0106-2.12 micrograms with the regression equation Y = 3035. 97 X - 14.85(r = 0.9999). The average recovery was 95.8% (n = 5, RSD = 1.3%).

CONCLUSIONThe method is simple, quick, sensitive and reproducible. In all of the samples, the calycosin-7-O-beta-D-glucopyranoside contents differ markedly.

Astragalus membranaceus ; chemistry ; classification ; China ; Chromatography, High Pressure Liquid ; Ecosystem ; Glucosides ; analysis ; Isoflavones ; analysis ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Species Specificity

The purpose of this study is to investigate the Sal I, Nru I and Mse I restriction fragment length polymorphisms (RFLPs) of factor IX gene in Chinese Han people. The frequencies of FIX-192 and FIX-793 for A and G, and FIX-698 for T and C were analyzed by polymerase chain reaction (PCR) in unrelated normal Chinese Han people. A sample of 214, 210 and 206 unrelated X chromosomes were analyzed for FIX-192 and FIX-793 and FIX-698, respectively. The results showed that the frequencies for FIX-192 were 0.878 for A and 0.122 for G, with a heterozygosity rate of 0.213, and the frequencies for FIX-793 were 0.552 for A and 0.448 for G, with a heterozygosity rate of 0.494, the frequencies for FIX-698 were 0.311 for T and 0.689 for C, with a heterozygosity rate of 0.429. It was concluded that the SalIand NruI and MseI RFLPs of FIX gene may be useful markers for carrier detection and prenatal diagnosis in Chinese families with hemophilia B patients.
China ; DNA ; genetics ; metabolism ; Deoxyribonucleases, Type II Site-Specific ; metabolism ; Factor IX ; genetics ; Female ; Gene Frequency ; Heterozygote ; Humans ; Male ; Polymorphism, Restriction Fragment Length

OBJECTIVETo obtain allelic loss mapping and define the minimal lost region on chromosome 1q21 in gastric carcinomas, and explore role of 1q21 loss of heterozygosity (LOH) in the development and progression of gastric carcinogenesis.

METHODSUsing 7 high-density microsatellite markers and PCR method, lq21 LOH was analyzed in 30 paired specimens of fresh gastric carcinoma, and the relation between 1q21 LOH and the clinicopathological features of the malignancy was tested.

RESULTSThe LOH frequency on chromosome 1q21 from these gastric carcinoma tissues reached 60% (18/30). The LOH frequencies of the microsatellite markers D1S514, D1S2696, D1S498, D1S305, D1S2624, D1S2635 and D1S2702 were 13.3%, 10%, 20%, 23.3%, 33.3%, 40% and 23.3%, respectively. The minimal lost region on 1q21 LOH in the gastric carcinoma tissues was located in the region between D1S2624 and D1S2707, in the vicinity of D1S2635. No significant relations of 1q21 LOH to the patients' age, gender, location of the primary foci, clinical staging, or the tumor differentiation were noted (P>0.05), but 1q21 LOH was correlated to lymph node metastases of the malignancy (P<0.05).

CONCLUSIONHigher frequency of 1q21 LOH occurs in gastric carcinoma cells, suggesting the presence of potential tumor suppressor genes closely associated with gastric carcinogenesis near the region of D1S2635.

Adenocarcinoma ; genetics ; Chromosome Mapping ; Chromosomes, Human, Pair 1 ; genetics ; Female ; Humans ; Loss of Heterozygosity ; Lymphatic Metastasis ; Male ; Microsatellite Repeats ; Middle Aged ; Stomach Neoplasms ; genetics

Objective To investigate the quality of life (QOL) and mental health status of patients with HIV/AIDS in Minhang District of Shanghai,and to explore the factors that affect their quality of life.Methods A cross-sectional study was conducted in Minhang District among HIV/AIDS patients.All subjects finished general situation questionnaire,Beck depression inventory (BDI),self rating anxiety scale (SAS),social support scale (SSS) and generic quality of life inventory-74 (GQOLI-74) survey.Results A total of 294 patients were recruited in this study with mean age of (39.6 ± 12.6) years old.Among the subjects,mean score of SAS was 40.5 ± 8.8,higher than national normative score (t =20.8,P＜0.001).The prevalence of anxiety was 13.9％.The mean score of BDI was 8.90 ± 8.59,and the prevalence of depression was 28.6％.The mean score of quality of life (QOL) was 66.6 ± 10.9.Multivariate logistic regression analysis showed that QOL of patients with high school or secondary school was lower than those with college and above education (OR =0.34,95 ％ CI:0.12-0.95).QOL scores of patients with moderate or severe depression were lower than those with less depressed patients,OR values were 0.14(95 ％ CI:0.06-0.34) and 0.07(95 CI:0.03-0.20)respectively.QOL of patients with anxiety was lower than the patients without anxiety (OR =0.10,95 ％CI:0.04-0.27).QOL of patients with high scores of social support was higher than the patients with low scores of social support (OR =3.95,95 ％ CI:1.82-8.59).Conclusions We should pay more attention to the psychological state of patients with HIV/AIDS.The quality of life can be improved by improving social support and reducing the occurrence of anxiety and depression.