Humans ; T-Lymphocytes, Cytotoxic ; immunology ; virology

Objective To explore the effect of perforin(PFP) antibody on the expression of Granzyme B (GzmB) in viral myocarditis.Methods Forty-five 4-week-old BALB/c male mice were randomly divided into 3 groups,including normal control group (n=15),viral control group (n=15) and PFP antibody therapy group (n=15).The mice in normal control group were inoculated with 0.15 mL Eagle reagent,and the mice in viral control group and PFP antibody therapy group were moculated with 0.15 mL TCID501012 L-1 coxsackievirus B3.The mice in PFP antibody therapy group were inoculated with PFP antibody (0.1 mg/kg) on 6 h and 3 d post inoculation.The mice in 3 groups were sacrificed on day 10 post inoculation.Extracted their hearts,then fixed,dehydrated,embeded and sliced the myocardial tissues.The expressions of GzmB proteins in myocardium were determined by immunohistochemistry.The areas of GzmB were measured by the Leica Qwin V3 system.SPSS 11.5 software was used to analyze the data.Results There were 4 mouse dead in viral control group,and 2 mouse dead in PFP autibody therapy group,no mice dead in normal contol group.The expressions of GzmB were not found in myocardial tissues of normal control group,but there were lots of expressions of GzmB in myocardial tissues of viral control group[(67.13?1.82)%].Athough the expressions of GzmB in PFP antibody therapy group[(6.98?1.34)%] were significantly decreased compared with those in myocardial tissues of viral control group(q=66.180 P

Objective To introduce the research progress of the modern methods in evaluating de qi sensation in acupuncture and moxibustion, and to analyze the current research situation and major problems. Method The objective evaluation methods majorly used to estimate qi sensation were summarized. Result The currently used evaluation methods basically include scales and cerebral function imaging, while the rest methods are still in the beginning stage. Conclusion As a subjective feeling, de qi sensation is difficult to determine and quantify objectively, and there lacks an objective standard. The study on the mechanism of de qi sensation is relatively insufficient and superficial.

According to foreign and domestic standards of clinical quality and the author team’ experience summary of long-term clinical study, it has been found that quality control in clinical acupuncture operators is closely related to operator’s ability, good communication between doctors and patients, signature for informed consent, implementation of inclusion and exclusion criteria, practice of blind method and random concealment, patient’s compliance, standardized writing and keeping of CRF, management of instruments and consumables, and coordination and association between researchers. The implementation method of quality control in clinical acupuncture operators is explored by the clinical practice of the author team in “973” project, and analysis and discussion of the above key points.

Objective:To establish an LC-MS/MS method for the simultaneous determination of acetaminophen and its five metabolites in mice plasma,and investigate the metabolism of acetaminophen by using the method.Methods:Para aminobenzoic acid was used as the internal standard.The plasma samples were precipitated by methanol,and then separated on a C18 column with the mobile phase of methanol and 5 mmol·L-1 ammonium acetate buffer solution containing 0.1% formic acid (55∶45).The flow rate was 0.5 ml·min-1,and the column temperature was 25℃.An electrospray ionization source was applied and operated in a positive ion mode using MRM:APAP,-,m/z 152.0→110.0;APAP-cys,-,m/z 271.2→140.1;APAP-glut,-,m/z 457.0→328.0;APAP-NAC,-,m/z 313.4 →208.0;APAP-sulf,-,m/z 232.4→152.1;APAP-gluc,-,m/z 328.2→152.1;IS,-,m/z 138.2→120.0.Results:The method exhibited good linearity over the concentration range of 0.2-10 μg·ml-1for APAP,1.0-20 μg·ml-1 for APAP-gluc,1.0-20 μg·ml-1 for APAP-sulf,1.0-20 μg·ml-1 for APAP-glut,0.4-15 μg·ml-1 for APAP-NAC and 0.2-10 μg·ml-1 for APAP-cys (r≥0.990 0).The inter-day accuracy and precision of acetaminophen and its five metabolites were all below 15%.The average recovery was between 85% and 115%,and RSDs were all below 15%.Conclusion:The LC-MS/MS method is proved to be quick,sensitive and accurate,and suitable for the determination of acetaminophen and its five metabolites in mice plasma.

Objective:To explore the histogenesis and differentiation of dermatofibroma (DF) and dermatofibrosarcoma protuberans (DFSP). Methods: Clinical information and microscopic characteristics of 26 cases of DF and 26 cases of DFSP were investigated. The immunohistochemical study was performed on microarray sections by a panel of antibodies including FactorⅩⅢa, HLA-DR, CD34, CD14, S-100, MSA, and Ki67. Probe was labeled by in vitro transcription. The mRNA expression levels of TGF-? and bFGF were investigated by in situ hybridization. Results: All cases showed positive for Factor ⅩⅢa,HLA-DR and CD34 to different extent. The medians of positive rates in DF were FactorⅩⅢa 90%, HLA-DR 70%, and CD34 5%, and in DFSP were FactorⅩⅢa 10%, HLA-DR 5%, and CD34 80%. CD14 was positive in 3 cases of DF and 1 case of DFSP. S-100 was positive in 6 cases of DFSP and 2 cases of DF. MSA was positive in 5 cases of DFSP and 3 cases of DF. In all cases, positive rate of Ki67 was less than 5%. The mRNA expression levels of TGF-? was elevated in DF in comparison with DFSP. Conclusion: Both DF and DFSP can differentiate to dendritic cells (DC) in different degree. Considering the character of microscopic features and immunohistochemical phenotype, cells of DF are much similar to mature DC, while those of DFSP much similar to immature dermal reserve cell (DRC). The differences of cell differentiation between DF and DFSP result in different prognosis. DF is a benign tumor, while DFSP a low grade malignant tumor. The different expression of FactorⅩⅢa and CD34 may be helpful to differential diagnosis of DF and DFSP.

According to the characteristic of teaching medicinal organic chemistry,it is helpful to discuss and advocate the application of ChemOffice software in teaching Medicinal Organic Chemistry so as to visualize the class teaching and enhance the efficiency of teaching.

In order to urge students majoring in medicine to know the basic knowledge of organic chemistry experiment in less lessons, enhance their abilities of self-study and train their abilities to think and solve problems, we tried the "PBL teaching method" flexibly during the experimental course teaching of "Medical Organic Chemistry". The result was better than usual.

To explore the inhibitory effect of hydroxyapatite (HA) particles with different sizes on malignant melanoma A375 cells in vitro, we synthesized 4 short rod-like HA particles using TIPS. Their mean diameters were 998.0 nm (HA1), 511.0 nm (HA2), 244.0 nm (HA3), and 71.6 nm (HA4), respectively. Malignant melanoma A375 cells were co-cultured with HA particles in vitro. Results showed that HA particles smaller than 511.0 nm in mean diameter could always inhibit proliferation of A375 cells, and nanometer-HA particles (HA4) had the strongest inhibitory effect on A375 cell proliferation and the strongest inducing effect on apoptosis. HA particles were distributed in plasma of A375 cells. The ultrastructure changes of A375 cells were found most significant in nanometer-HA particles (HA4) group. We conclude that particle size is a very important influencing factor on anti-tumor effects of HA and that nanometer-HA particle has the strongest inhibitory effect on tumor cell proliferation.
Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Durapatite ; chemistry ; Humans ; Melanoma ; Nanotubes ; chemistry ; Particle Size

Background Ginkgolide B (GB) has been proved to have neuroprotective and anti-apoptotic effects and can effectively inhibit apoptosis of retinal photoreceptor cells.But the high hydrophobic feature and low bioavailability of GB limit its clinical application.Self microemulsifying drug delivery system (SMEDDS) can effectively improve the infusibility drug dissolution and bioavailability in the retina.Objective This study was to investigate the pharmacokinetics and drug-time change of GB-loaded SMEDDS in retina.Methods Eighty SD rats were randomized into 2 groups,2.5％ GB(40 mg/kg) of SMEDDS or GB suspension(0.1％ DMSO dissolve) were gastrically given respectively in two groups.The rats were sacrificed and retinas were isolated 15,30,45 minutes and 1 hour,2,4,8,12 hours to prepare the retinal suspension.The content of GB in retina was assayed with high performance liquid chromatography-electrospray ionization-(1) (1)ss spectrum (HPLC-ESI-MS) and contrasted with standard curve.Practical drug dynamics program 3p87 was used to detect the pharmacokinetics parameters.The maximal content(Cmax,mg/g),time to peak (Tmax,h),clearance ratio (Ke/h),high-life period (t1/2) and area under the concentration-time curve(AUC0-∞,mg/(g · h)) of GB in various time points in retina after a single oral dose were calculated and compared between two groups.Results The standard curve was obtained over the concentration range of 1-32 mg/L with a linear regression equation,Y =0.0732X + 0.056 (r =0.992).A similar content-time curve was seen between GB suspension group and GB-SMEDDS group.The GB content was higher in GB-SMEDDS group than that in GB suspension group from 30 minutes through 12 hours after administration of drugs.The Cmax of GB-SMEDDS group and GB suspension group were(15.83±1.84) mg/g and(2.65±0.10) mg/g,the AUC0-∞ were(15.30±0.11)mg/(g· h)and(6.42±0.19)mg/(g · h).Conclusions HPLC-ESI-MS is proved to be a rapid,accurate,sensitive and suitable method for pharmocokinetic study of GB.SMEDDS can raise the concent of GB in retina,and it probably improve the bioavailability of GB.