Genetic mutations are important molecular biomarkers for cancer diagnosis and surveillance.Therefore,the development of methods for mutation detection characterized with straightforward,highly specific and sensitive to low-level mutations within various sequence contexts is extremely needed.Although some of the currently available methods have shown very encouraging results,their discrimination efficiency is still very low.Herein,we demonstrate a fluorescent probe coupled with blocker and property of melting temperature discrimination,which is able to identify the presence of known or unknown single-base variations at abundances down to 0.1％ within 20 min.The discrimination factors between the perfect-match target and single-base mismatched target are determined to be 10.15-38.48.The method is sequence independent,which assures a wide range of application.The new method would be an ideal choice for high-throughput in vitro diagnosis and precise clinical treatment.

Background The domestic and international researches discovered that many proteins and enzymes of the extracellular matrix (ECM) participate in the sclera remodeling by affecting the collagen typeⅠand fibronectin.Objective This study was to investigate the effect of matrixmetalloproteinase-2 (TIMP-2) on expression of collagen typeⅠand fibronectin of ECM in the posterior sclera by injecting liposomes containing tissue inhibitor of TIMP-2 gene into suprachoroidal space of the form-deprivation myopia in guinea pig.Methods Form-deprivation myopia was induced by translucent goggles in 36 clean guinea pig for 2 weeks.Then the animals were randomly assigned to TIMP-2 group,empty plasmid group,saline group and 12 for each group.Liposomes of 5μl containing TIMP-2 gene,empty plasmid and saline were suprachoroidally injected in the right eye respectively,and the left eyes without any treatment were used as self-control group.Other 12 matched guinea pigs only covered the right eyes through out the experimental duration as model control group.The guinea pigs were sacrificed and the posterior sclera tissue of the eyeballs were collected at 2,7 and 14 days after injection of drug.The expressions of collagen typeⅠmRNA and fibronectin mRNA were detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR).This study followed the Regulation for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results The expression level of collagen type Ⅰ mRNA in the posterior sclera of guinea pig was lower but that of fibronectin mRNA was higher in TIMP-2 group than self-control group,showing significant differences between them (P＜0.05).The expression level of collagen type Ⅰ mRNA in the posterior scleral tissue began to increase from the 2nd day after drug injection and was obviously elevated at the 7th day and then gradually decreased at the 14th day.However,the expression level of fibronectin mRNA in the posterior scleral tissue showed the opposite pattern.The expression levels of collagen typeⅠmRNA and fibronectin mRNA at the 7th after drug injection were significantly lower than that at the 2nd day or 14th day (P＜0.01).Conclusion Suprachoroidal injection of TIMP-2 in form-deprivation myopia could up-regulate the expression of collagen typeⅠmRNA and down-regulate the expression of fibronectin mRNA in the posterior scleral tissue.It may slow down the sclera remodeling of form-deprivation myopia in guinea pig in the early stage.

High-speed counter-current chromatography (HSCCC) was used to high performance separate and prepare lignans from Schisandrae chinensis fructus. The solvent system is composed of n-hexane-ethyl acetate-methanol-water (9 : 1 : 5 : 5) and n-hexane-ethyl acetate-methanol-water (9 : 1 : 9 : 5), speed is at 900 r.min-1, and flow rate is at 2.0 mL.min-1. Five fractions from Schisandrae chinensis fructus extract were separated and prepared with one HSCCC process. They were identified as schisandrin, gomisin J, schisandrol B, schisantherin A and deoxyschizandrin by electrospray ionization-multiple tandem mass spectrometry (ESI-MSn), respectively. Their contents were obtained in 98.74%, 94.32%, 99.53%, 94.23% and 98.68% by ultra high performance liquid chromatography (UPLC), separately. The rapid and simple method can be applied for the preparation of lignans from Schisandrae chinensis fructus.
Countercurrent Distribution ; Cyclooctanes ; chemistry ; isolation & purification ; Dioxoles ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Fruit ; chemistry ; Lignans ; chemistry ; isolation & purification ; Molecular Structure ; Plants, Medicinal ; chemistry ; Polycyclic Compounds ; chemistry ; isolation & purification ; Schisandra ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry

OBJECTIVETo observe the effect of Chinese medicine and pharmacy (CMP) on the mortality of senile HIV/AIDS patients as adjunctive therapy.

METHODSHIV/AIDS patients of a certain rural area of Hanna Province, who were recruited in national CMP HIV treatment trial program (NTCMTP) in 2004, were enrolled as the CMP treatment group. HIV/AIDS patients in the same village without recruiting in NTCMTP were enrolled as the non-CMP treatment group. Data related to subjects were collected from the database of NTCMTP and National HAART Reporting System. Multiple regression analysis under Cox proportional hazard model was applied to examine the risk factors for death of senile HIV/AIDS patients.

RESULTSA total of 436 HIV/AIDS were enrolled in this study, 204 in the CMP treatment group and 232 in the non-CMP treatment group. There were 70 AIDS-relative deaths in the CMP treatment group, with 8-year mortality rate of 37.74%. There were 111 AIDS-relative deaths in the non-CMP treatment group, with 8-year mortality rate of 48.34%. The 8-year mortality rate was higher in the non-CMP treatment group than in the CMP treatment group (chi2 = 5.136, P < 0.05). Results of univariate Cox proportional hazards regression analysis showed the hazard ratio in the non-CMP treatment group was 1.41 times that of the CMP treatment group (P < 0.05). Result of multivariate Cox proportional hazards regression analysis showed the hazard ratio in the non-CMP treatment group was 1.44 times that of the CMP treatment group (P < 0.05). Besides, gender and marital conditions were significantly associated with death of HIV/AIDS patients.

CONCLUSIONCMP treatment was favorable to lower the mortality rate of senile HIV/AIDS patients, and its objective evaluation awaits for further prospective study.

Acquired Immunodeficiency Syndrome ; drug therapy ; mortality ; Alzheimer Disease ; therapy ; Antiretroviral Therapy, Highly Active ; Communicable Diseases ; Drugs, Chinese Herbal ; therapeutic use ; HIV Infections ; drug therapy ; mortality ; Humans ; Proportional Hazards Models ; Prospective Studies ; Risk Factors

Objective To detect the expression of metallothionein-3 (MT-3)mRNA in human esophageal squamous cell carcinoma. Methods Five cell lines of human esophageal cancer,TE-1,TE-13,TTN,ECA-109 (cell lines of esophageal squamous cell carcinoma) and OE33 (cell lines of esophageal adenocarcinoma),were used in this study. RT-PCR was employed to detect the expression of MT-3 mRNA. Peripheral blood monouclear cells from normal subjects were served as controls. Results Sequencing of RT-PCR product certified the gene of MT-3 mRNA. It was revealed by gel electrophoresis that there was expression of MT-3 mRNA in each cell line. The relative expression of MT-3 mRNA was 0.230?0.023,0.516?0.020,0.140?0.009,0.176?0.015 and 0.085?0.011 in cell lines of TE-1,TE-13,TTN,ECA-109 and OE33,respectively,significantly lower than that in controls (0.762?0.026) (P

Objective To synthesize 99Tcm- (hydrazinonictinamide- [Lys3] -bombesin) (tricine)(trisodium triphenylphosphine-3,3',3"-trisulfonate) ((HYNIC-[Lys3]-BBS) (tricine) (TPPTS)) and evaluate its biodistribution and binding capability with tumor tissue in Balb/c nude mice bearing human pancreatic cancer xenografts. Methods HYNIC was conjugated to the [Lys3] -BBS at pH = 9.0 with SnCl2 as reducing agent and both tricine and TPPTS as coligands for 99Tcm-labeling. 99Tcm-HYNIC-[Lys3]-BBS)(tricine) (TPPTS) was purified by Sep-Pak C18 cartridge and was analysed by HPLC. The radiochemical purity and radiolabeling yield were measured. The stability of 99Tcm-(HYNIC-[Lys3]-BBS) (tricine)(TPPTS) in serum, biodistribution (% ID/g) in the normal mice and imaging of the Balb/c nude mice bearing human pancreatic cancer xenografts in vivo were studied. Results The radiolabeling yield was (90 ±2)% and the radiochemical purity was over 95%. The radiochemical purity after 4 h in serum was over 85%. The distribution in normal mice showed rapid clearance from blood (the uptake was (0.07 ±0.01) %ID/g at 2 h postinjection). 99Tcm-(HYNIC-[Lys3]-BBS) (tricine) (TPPTS) was excreted mainly via the kidney with little radioactivity accumulation in the liver and gastrointestinal tract (the uptake of liver, stomach, intestine was (0.27 ±0.03), (0.06 ±0.03), (0.04 ±0.00) %ID/g at 2 h postinjection). Marked uptake of radioactivity was found in tumor tissue of the Balb/c nude mice bearing human pancreatic cancer with maximum T/NT ratio of 3.71 ± 0.57 at 2 h postinjection. Conclusions 99Tcm-(HYNIC-[Lys3]-BBS)(tricine) (TPPTS) can be easily prepared with high radiolabeling yield and radiochemical purity. The stability in serum and good biodistribution charateristics make it useful for the diagnosis of human pancreatic cancer with over-expression of the gastric-releasing peptide(GRP) receptor.

Animals ; Insecticides ; toxicity ; Nitric Oxide ; blood ; Organophosphorus Compounds ; Rabbits ; Superoxide Dismutase ; blood ; Vitamin E ; blood

Objective To explore the effect of sleep deprivation (SD) on cognitive evaluation of affective picture. Methods Korty-three undergraduates were recruited and assigned to sleep deprivation group (n 23) and sleep control group (n=20). Because two students in the sleep control group did not participate in retest task and one student data of the test were bst. 17 people was eventually included in the sleep control group. The students in the sleep deprivation group received sleep deprivation at the end of the test (from 22:00 to next day 8:00). The sleep control group had no intervention. A total of 206 affective pictures (108 test and 108 retest) were selected from International Affective Picture System (1APS) and categorized as positive, neutral and negative pictures. The Positive and Negative Affect Scale ( PANAS) was used to investigate the subjective mood ratings of participants at test and retest in two groups and to evaluate the effect of SD on cognitive evaluation of affective pictures. Results SD showed no significant effect on the evaluation of positive and negative pictures, but it showed a negative bias in valence ratings for neutral pictures, with significant difference found for neutral pictures between test and retest in sleep deprivation group (P<0. 01). but not in the sleep control group (P= 0.12). After controlling covariance subjective emotion, the negative bias caused by SD still existed for the neutral pictures. The arousal ratings for affective pictures in sleep deprivation group was significantly higher than that in sleep control group (P<0.05). Conclusion Our results indicate that sleep is important in emotional evaluation, and SD can lead to a negative bias for neutral stimuli.

Sleep is very important for maintaining normal emotional function. Sleep deprivation has been shown to decrease the individual positive emotion and enhance the level of negative ones. This paper reviews the current findings with regard to the effects of sleep deprivation on emotional state, hoping to better understand the underlying mechanisms by which sleep deprivation affects individual emotion from the following perspectives:emotional brain networks, rapid eye movement sleep, emotional information processing, energy supplying and emotional background.

Objective To investigate the expression of peroxisome proliferators-activated receptor ? (PPAR?) in trophoblast and relation between PPAR? ligands and trophoblast invasion.Methods We examined the expression of PPAR? by immunohistochemistry,immunocytochemistry and real time quantitative PCR.We next examined,using the cytotrophoblast culture model,the biological role of PPAR? ligands in vitro.Results PPAR? was mainly localized in the nuclei of villous cytotrophoblast and extravillous cytotrophoblast of cell islands and cell columns.In villous tissue and cultured trophoblast from early first trimester,the level of expression of PPAR? mRNA and protein was 36.0?5.1,13.4?3.1 and 1.35?0.08,1.13?0.11;from late first trimester it was 23.3?5.5,6.1?1.3 and 1.17?0.03,0.86 ?0.05,and the expression of PPAR? was obviously decreased (P