Objective To observe the change of Toxoplasma g ondii tachyziote in pseudocyst. Methods The mice infected by RH Toxoplasma were treated with mandelic acid (200 mg?kg -1 , twice daily) orally or intravenously, then ascites was taken out to smear in 24 h, 72 h and the time of death, with Giemsa dye and transmission electronic microscope (TEM), the survival time was c alculated. Results Under the light microscope, the cell mem brane of tachyzoite was tortous and broken, the bubble and pelletish material we re observed in cytoplasm, cell nucleus was split; but the cell membrane, organs and nucleus were destroyed more obviously under the TEM than those under the lig ht microscope. Meanwhile the living time of mice treated by mandelic acid (8.0 d ays in oral administration group and 6.8 days in intravenous administration grou p) was obviously longer than that in positive control group(5.5 days, P

Objective:To simultaneously determine the contents of asarinin, prim-O-glucosylcimifugin and 5-O-methylvisammio-side in Xinqin granules. Methods:An HPLC method was used. The determination was performed on a ZORBAX Eclipse XDB-C18 col-umn(150 mm ×4.6mm,5 μm) with the mobile phase consisting of menthol (A)-water (B) with gradient elution. The flow rate was 1. 0 ml·min-1 . The column temperature was 30℃. The detection wavelength was set at 254 nm from 0 to 30 min and 287nm from 30 to 55 min. The injection volume was 10μl. Results:The linear range of prim-O-glucosylcimifugin, asarinin and 5-O-methylvisammio-side was 10.210-163.400 μg·ml-1(r=0.999 7),10.160-162.600 μg·ml-1(r=0.999 8) and 5.015-80.240 μg·ml-1(r=0. 999 8), respectively. The average recovery was 100. 30%(RSD=1. 6%, n=6),101. 53%(RSD=1. 1%,n=6) and 101. 12%(RSD=1. 2%, n=6), respectively. Conclusion: The method is simple and accurate, which can be used in the quality control of Xinqin granules.

To develop a LC-MS/MS method for the determination of five kinds of trace ginkgolic acids in diterpene ginkgolides meglumine injection materials, the column was Agilent ZORBAX Eclipse plus C18 (3.0 mm x 50 mm, 1.8 µm), and the mobile phase consisted of methanol-water (containing 0.2% formic acid) (95:5) at a flow rate of 0.5 mL · min(-1). The multiple reaction ion monitoring (MRM) with an ESI interface in the negative ion mode was selected. The results showed that the linear ranges of five kinds of ginkgolic acids were in the range of 0.2-36.0 µg · L(-1) (r ≥ 0.999 5). The lowest limit of quantification (LOQ) of ginkgo acid C13: 0, C15:1, C17:2, C15:0 and C17:1 were 0.18, 0.18, 0.21, 0.10 and 0.20 µg · L(-1), respectively. The average recovery was between 73.28% and 87.56%, and the average content of total ginkgolic acids in three batches of samples was in the range of 0.023-0.028 µg · g(-1), which was much lower than 2 µg · g(-1) prescribed in drug registration standards. This method is simple and rapid with high sensitivity, which can be used for the determination of five kinds of trace ginkgolic acids in diterpene ginkgolides meglumine injection materials.
Chromatography, Liquid ; methods ; Ginkgolides ; analysis ; Injections ; Limit of Detection ; Salicylates ; analysis ; Tandem Mass Spectrometry ; methods

The changes of microRNA expression in rat hippocampus after traumatic brain injury (TBI) were explored. Adult SD rats received a single controlled cortical impact injury, and the ipsilateral hippocampus was harvested for the subsequent microarray assay at three time points after TBI: 1st day, 3rd day and 5th day, respectively. We characterized the microRNA expression profile in rat hippocampus using the microRNA microarray analysis, and further verified microarray results of miR-142-3p and miR-221 using quantitative real-time PCR. Totally 205 microRNAs were identified and up-/down-regulated more than 1.5 times. There were significant changes in 17 microRNAs at all three time points post-TBI. The quantitative real-time PCR results of miR-142-3p and miR-221 indicated good consistency with the results of the microarray method. MicroRNAs altered at different time points post-TBI. MiR-142-3p and miR-221 may be used as potentially biological markers for TBI assessment in forensic practice.

BACKGROUND:To date,no single material can completely meet the clinical requirements.However,the composite materials characterized by good biodegradability,biocompatibility and osteoconductivity have become a highlight of the artificial bone materials.OBJECTIVE:To synthesize the basic fibroblast growth factor (bFGF)-poly(lactic-co-glycolic acid) (PLGA) microspheres/hydroxyapatite (HA)/poly(I-lactic acid) (PLLA) porous bone scaffolds,and to observe the physicochemical properties and biocompatibility of the composite material.METHODS:The bFGF-PLGA microspheres were prepared by double emulsion method,and then six kinds of materials were made including PLLA,PLLNHA,PLLAJPLGA,PLLNHNPLGA,PLLNHA/bFGF,and bFGF-PLGA microspheres/PLLA/HA.The characterization of the materials were observed by particle size analyzer,transmission electron microscopy,X-ray diffraction,Fourier transform infrared spectrometer,microcomputer differential thermal balance,and scanning electron microscope.Toxicity of these materials and proliferation of bone marrow mesechymal stem cells seeded onto these materials were analyzed and compared.RESULTS AND CONCLUSION:The average particle size of bFGF-PLGA microspheres was about 250 nm,the average drug-loading capacity was (26.03±0.17)％,and the entrapment percentage was (90.65±2.68)％.The prepared bFGF-PLGA microspheres were spherical and had good dispersibility.In addition,all the six kinds of materials had a porous structure with similar pore diameter,in which the microspheres and particles exhibited a rational distribution.The toxic level of bFGF-PLGA microspheres/PLLNHA,bFGF/HNPLLA and HAP/PLLA was graded as 1 (with a relative survival rate ≥ 80％),indicating no obvious toxicity or slight toxicity.All these six kinds of composite materials can promote the proliferation of bone marrow mesenchymal stem cells,and the bFGF-PLGA microspheres/PLLNHA shows the best effects on cell proliferation and has good biocompatibility.

Objective:To investigate the potential anti-aging mechanism of 9-hydroxy-6,7-dimethoxydalbergiquinol (HDDQ) on hydrogen peroxide (H2O2)-induced oxidative stress in human dermal fibroblasts (HDFs). Methods:The effect of HDDQ on cell viability was assessed by MTT assay, and the effects of HDDQ on senescence-like phenotypes were determined by senescence-associated β-galactosidase (SA-β-gal) staining, Western blotting analysis, and a cell proliferation assay. The expression level and activity of sirtuin-1 (SIRT1) induced by HDDQ were also measured. Results:HDDQ reversed senescence-like phenotypes in the oxidant-challenged model, through reducing SA-β-gal activity and promoting cell growth. Meanwhile, decreases in ac-p53, p21Cip1/WAF1, and p16Ink4a and an increase in pRb were observed. HDDQ induced the expression of SIRT1 in a concentration- and time-dependent manner. Moreover, HDDQ inhibited H2O2-induced phosphorylation of Akt by SIRT1 up-regulation and reduced SA-β-gal staining. Conclusions:HDDQ inhibits H2O2-induced premature senescence and upregulation of SIRT1 expression plays a vital role in the inhibition of the senescence phenotype in HDFs.

Luteolin, a sort of flavonoid, has been reported to be involved in neuroprotective function via suppression of neuroinflammation. In this study, we investigated the protective effect of luteolin against oxidative stress-induced cellular senescence and its molecular mechanism using hydrogen peroxide (H2O2)-induced cellular senescence model in House Ear Institute-Organ of Corti 1 cells (HEI-OC1). Our results showed that luteolin attenuated senescent phenotypes including alterations of morphology, cell proliferation, senescence-associated β-galactosidase expression, DNA damage, as well as related molecules expression such as p53 and p21 in the oxidant challenged model. Interestingly, we found that luteolin induces expression of sirtuin 1 in dose- and time-dependent manners and it has protective role against H2O2 -induced cellular senescence by upregulation of sirtuin 1 (SIRT1). In contrast, the inhibitory effect of luteolin on cellular senescence under oxidative stress was abolished by silencing of SIRT1. This study indicates that luteolin effectively protects against oxidative stress-induced cellular senescence through p53 and SIRT1. These results suggest that luteolin possesses therapeutic potentials against age-related hearing loss that are induced by oxidative stress.

Luteolin, a sort of flavonoid, has been reported to be involved in neuroprotective function via suppression of neuroinflammation. In this study, we investigated the protective effect of luteolin against oxidative stress-induced cellular senescence and its molecular mechanism using hydrogen peroxide (H2O2)-induced cellular senescence model in House Ear Institute-Organ of Corti 1 cells (HEI-OC1). Our results showed that luteolin attenuated senescent phenotypes including alterations of morphology, cell proliferation, senescence-associated β-galactosidase expression, DNA damage, as well as related molecules expression such as p53 and p21 in the oxidant challenged model. Interestingly, we found that luteolin induces expression of sirtuin 1 in dose- and time-dependent manners and it has protective role against H2O2 -induced cellular senescence by upregulation of sirtuin 1 (SIRT1). In contrast, the inhibitory effect of luteolin on cellular senescence under oxidative stress was abolished by silencing of SIRT1. This study indicates that luteolin effectively protects against oxidative stress-induced cellular senescence through p53 and SIRT1. These results suggest that luteolin possesses therapeutic potentials against age-related hearing loss that are induced by oxidative stress.

Objective To explore the effect of propofol on the apoptosis of PC12 cell induced by glutamic acid.Methods PC12 cells were in-duced 10 mmol · L-1 glutamate for 48 h, and were divided into model group , propofol low , medium and high dose groups.The normal cells were used as control group.Control group and model group were received culture medium without any drugs for 48 h.Propofol low , medium and high dose groups were given 12.5 , 25.0 , 50.0 μmol · L-1 propofol for 48 h.The viability of PC12 cell was measured by MTT assay.PC12 cell apoptosis was measured by flow cytometry.The activity of Caspase -3 was determined by spectrophotometric method.The expression of FBJ osteosarcoma oncogene ( c -fos ) and early growth response protein 1 ( Egr -1 ) were detected by Realtime -PCR and western blot.Results Compared with the normal group , the cell apoptosis rate , the activity of Caspase -3 , the expression of c -fos mRNA and protein increased , and the cell viability , the expression of Egr -1 mRNA and protein decreased in the model group ( P<0.01 ).Compared with the model group, the acitivity of Caspase -3, the cell apoptosis rate , the activity of Caspase -3, the expression of c-fos mRNA and protein decreased , and the cell viability , the expression of Egr-1 mRNA and protein increased in propofol low -dose , medium -dose and high -dose group ( P <0.05 ).Conclusion Propofol suppressed the apoptosis of PC 12 cell induced by glutamic acid , which was related with the expression of c-fos and Egr-1.

OBJECTIVETo establish a cohort of human immunodeficiency virus (HIV) discordant couples for follow-up studies and to collect data on frequency of HIV heterosexual transmission and related factors.

METHODSA total of 52 HIV discordant couples were identified by face to face interview and serological testing, in which the HIV negative individuals had no HIV infection behaviors including injecting drug use, blood transfusion or having sexual partners other than his/her own wife/husband. Three times of follows-up studies were carried out in 0.5 year, 1 year and 2.5 years to collect information on their sexual practices and condom use through face to face interview together with 20 ml whole blood collected to test HIV antibody, CD4+ T cell count and viral load.

RESULTS(1) In the period of 2.5 years follow-up, no HIV seroconversion and HIV transmission was found. (2) The frequencies of sexual intercourse between once per month to once per week were 65.4%, 72.9%, 71.7% and 80.0% at the time of cohort setup: 0.5 year, 1 year and 2.5 years of follow-up respectively. The rates of "occasional use" to "never use" condoms were 76.9%, 66.6%, 69.1% and 60.0% at the time of cohort setup as: 0.5 year, 1 year and 2.5 years of follow-up, respectively. No significant difference between different times of follow-up for sexual intercourse or condom use. (3) 85.4%, 66.6% and 60.0% of the HIV positive individuals kept their CD4+ T cell count stabilized or raised during the 0.5 year, 1 year and 2.5 years follow-up period, respectively. However, 66.7% of them showed stable or declined viral load in the period of 2.5 years follow-up. It appeared that stable or raised CD4+ T cell and the stable/declined viral load happened simultaneously.

CONCLUSIONNo transmission was identified in this study. The stabilized CD4+ T cell count and viral load might be account for the reason of no transmission while the biological factors from host and virus related with transmission need to be further studied.

CD4 Lymphocyte Count ; China ; epidemiology ; Cohort Studies ; Coitus ; Condoms ; Contraception Behavior ; Female ; HIV ; physiology ; HIV Infections ; epidemiology ; immunology ; transmission ; virology ; Humans ; Male ; Rural Health ; Spouses ; Viral Load