Objective To study CD40 expression on melanocytes induced by IFN-?and its significance. Methods CD40 expression was detected by flow cytometry. The capacity of melanocytes to stimulate T lymphocytes was evaluated by mixed ly mphocyte reaction and the supernatant cytokine levels were determined by ELISA. ResultsHuman melanocytes(MC) cultured in vitro expressed low but detectable CD40 surf ace protein. The surface expression of CD40 was markedly up-regulated by stimul ation with interferon(IFN)-?with different concentrations for 24 hours,48 hour s and 72 hours, respectively. The expression of CD40 was correlated with IFN-? levels after 24 hour incubation. MC underwent a morphologic change with an incre ased capacity to stimulate allogenic lymphocytes to proliferate after IFN-?sti mulation. Optimal enhancement of stimulating index(SI) was observed at an IFN-?concentration of 300 IU/ml after 72 hour treatment. Meanwhile concentrations o f interleukin-12 but not interleukin-8 or 10 were obviously increased in the s upernatants of cultured MC. Furthermore, ligation of CD40 via soluble CD40 ligan d(SCD40L) could enhance CD80 and ICAM-1 expression, which could be blocked by s pecific monoclonal antibody to CD40L. Conclusions Since CD40-CD40L is a pair of important and special costimulating signal, it is of great value to elucidate t he fact that CD40 is functionally expressed on MC, thus for a better understandi ng of MC′s role in cellular immune responses. MC might activate cytotoxic T lym phocyte directly and not via CD4 positive lymphocyte.

OBJECTIVE: To optimize the process of extacting effective constituents from aerial part of Rheum officinale Baill by Plackett-Burman design combined with CCD response surface methodology. METHODS: In Plackett-Burman combined with CCD response surface design, the independent variables were concentration of ethanol, extraction time, and solvent fold. The dependent variables were contents of total anthraquinone and rheum emodin. RESULTS: The optimal extraction process of total anthraquinone was as follows: 67.25% ethanol, 90 min for reflux per time, 28:1 fold of solvent and 2 times of extraction; the optimal extraction process of emodin was 85% ethanol, 90 min for reflux per time, 40 fold of solvent and 2 times of extration. CONCLUSION: The optimal process is simpleand convenient for extracting aerial part of Rheum officinale Baill with high precision and predictability. Copyright 2012 by the Chinese Pharmaceutical Association.

Objective: To establish a method for the quality control of the extracts from the aerial parts of Rheum officinale (EARO) based on multi-assay depending on one determination (MDOD). Methods: The quality control of EARO was carried out through the thin-layer chromatography(TLC), discrimination of chromatographic peak in fingerprint, ultraviolet spectrophotometric, and HPLC methods. The relative correction factors of aloe-emodin, chrysophanol, and physcione for detecting emodin were set up, the factors were used to measure their contents, and the method was adopted to the quality control of EARO. Results: The experimental and control samples in TLC showed the same colored spots at the same position in EARO, five characteristic peaks among six were identified, and showed a good linear relationship at the range of 4.944-29.664 μg/mL (r = 0.9990). The method of MDOD showed the recoveries of aloe-emodin, emodin, chrysophanol, and physcione were 97.08%, 95.78%, 100.60%, and 97.47%, and the RSD values were 3.32%, 2.42%, 3.72%, and 2.67%. There were no significant differences between MDOD method and external standard method. Conclusion: TLC and fingerprint could be used to identify the chemical constituents in EARO, and ultraviolet spectrophotometry could be used to control the quantity of the total anthraquinone. The method of MDOD could be used to quantitatively control the contents of aloe-emodin, emodin, chrysophanol, and physcione, and the method is simple and accurate, which has a higher repeatability.

A novel simple,sensitive fluorescence immunosensing method based on aptamer-plasmid complex amplification was developed. This method utilized the specific recognition between antibody and antigen as well as aptamer-plasmid complex and the intercalation of fluorescence dye SYBR Green Ⅰ in the groove of duplex plasmid DNA in detection of Platelet-Derived Growth Factor BB (PDGF-BB). The immunoassay was performed in the microtiter wells in which rabbit anti PDGF-BB antibody was immobilized. The PDGF BB analyte was captured by the primary antibody and then sandwiched by the aptamer-plasmid DNA complex. The introduction of fluorescence dye SYBR Green Ⅰ allows for the detection of the sandwiched immunocomplex of antibody/anigen/aptamer-plasmid complex. Under the optimized conditions of salt concentration,ratio of aptamer to PUC19,and SYBR Green Ⅰ concentration,the proposed method offers a linear detection range from 0.2 μg/L to 200 μg/L with a detection limit of 0.1μg/L.

Objective To investigate the changes of serum soluble factor-related apoptosis (sFas) and soluble Fas ligand (sFasL) and their relations with cognition disorders in the patients with vascular dementia (VaD). Methods Serum concentrations of sFas and sFasl were detected by enzyme-linked immunosorbent assay (ELISA) and compared between 70 patients with VaD aged (72.5± 7.5)years and 50 healthy elderly people aged(72.5 ± 7.5)years.The VaD patient's cognitive functions were evaluated by activity of daily living scale (ADL),mini mental state examination (MMSE) and hachinski ischemia score (Hachinski). Results The serum levels of sFas and sFasL in VaD patients were (228.0±60.7)μg/L and (146.8±30.1)μg/L,and in the healthy elderly were (62.4±22.6)μg/L and (82.3 ± 18.7)μg/L,respectively.The serum levels of apoptosis factors in VaD patients were significantly higher than in the healthy controls (t=20.883,14.453,P＜0.01).sFas level was negatively correlated with age,the scores of ADL and Hachiuski while positively with the scores of MMSE (r=-0.956,-0.943,-0.950 and 0.904,all P＜0.01). sFasL level was negatively correlated with the scores of MMSE while positively with age,the scores of ADL and Hachinski (r=-0.899,0.963,0.948 and 0.939,a11 P＜0.01). Conclusions Apoptosis may be involved in the pathological change during VaD and the serum levels of sFas and sFasL might be related with cognition disorders.

The authors of this paper confirm the goal, factors and principles in arrangement of clinical practice, design a transplacement algorithm to solve the model, and after estimating the capacity to interns of affiliated hospital, make the schedule by that algorithm so as to relieve the relative short supply of good clinical education resourses.

Objective To investigate the effects of lipid-associated membrane proteins ( LAMPs) derived from Mycoplasma pneumoniae ( M.pneumoniae) strains on the expression of heme oxygenase 1 ( HO-1) in a human monocyte cell line (THP-1).Methods THP-1 cells were in vitro cultured with different concentrations of LAMPs for different times.The cytotoxicity of LAMPs to THP-1 cells was analyzed by using lactate dehydrogenase ( LDH) releasing test.The expression of HO-1 at protein and mRNA levels were de-tected by Western blot and real-time RT-PCR, respectively.The enzymatic activity of HO-1 protein was ex-amined by colorimetric assay.THP-1 cells stimulated with PBS and LPS were set up as the negative and pos-itive controls, respectively.Results A significantly enhanced LDH releasing rate was observed in THP-1 cells treated with 10 μg/ml of LAMPs.The expression of HO-1 at protein and mRNA levels in THP-1 cells were induced by LAMPs in a dose-dependent and time-dependent manner.The highest level of HO-1 protein was detected in THP-1 cells treated with 5.0 μg/ml of LAMPs.The transcriptional levels of HO-1 induced by LAMPs were significantly elevated at 3 h, peaked at 9 h and were decreased at 12 h.The expression of HO-1 protein in THP-1 cells was enhanced after 8 h of treatment with LAMPs and a significant decrease was observed at 20 h after reaching peaks at 12 h and 16 h.The activity of HO-1 protein was significantly en-hanced along with the increased expression of HO-1 protein.Conclusion The LAMPs derived from M.pneumoniae strains induced the expression of HO-1 at mRNA and protein levels.Moreover, the enzyme activity of HO-1 protein was enhanced in LAMPs treated THP-1 cells.

Objective To establish a CVS-11 pseudovirus particles ( pp)-based assay for detec-tion of neutralizing antibody against rabies virus. Methods An improved rapid fluorescence focus inhibition test ( RFFIT) for detection of neutralizing antibody against rabies virus ( RVNA) was established based on the CVS-11 pseudovirus expressing a luciferase reporter gene. Forty-six human serum samples were analyzed with the improved RFFIT and the results were compared with those by using standard RFFIT. Moreover, the improved RFFIT was used to detect the titers of RVNA in 91 serum samples collected from pet dogs and pet-breeders in Beijing. Results The coincidence rate of the improved RFFIT and the standard RFFIT was 100% regarding to the analysis of 46 human serum samples and 5 negative reference serum samples. Moreo-ver, the RVNA titers of all serum samples obtained with CVS-11 pseudovirus-based assay showed a signifi-cant high correlation with those obtained with standard RFFIT (n=46, r=0. 94, P<0. 000 1). All of the 91 serum samples collected from pet dogs and pet-breeders in Beijing were positive for RVNA as indicated by the improved RFFIT with a mean titer of 33. 01 IU/ml. Conclusion We established an improved RFFIT based on the CVS-11 pp expressing luciferase reporter gene, which might be used as a reliable alternative RFFIT for measuring RVNA titer. Analysis of the 91 serum samples collected in Beijing with the improved RFFIT showed that all samples were positive for RVNA.

Objective To explore the factors of relapse in alcohol-induced psychiatric and behavioral disorders.Methods103 male inpatients met with the diagnostic criteria of alcohol-induced psychiatric and behavioral disorders according to ICD-10 were enrolled.All patients were hospitalized from Wuxi Mental Center from January 2013 to August 2015.As baseline,information was obtained within all patients,and relapse was evaluated one year after discharge.The Kaplan-Meier method was used to statistically analyze the alcohol relapse time of patients with different length of hospitalization.Cox regression was used to explore the risk factors for alcohol relapse,including age,education level,marital status,family history,smoking,fixed income,number of hospitalizations,duration of alcohol intake,average daily alcohol intake,time of psychosis,psychiatric symptoms,length of hospitalization,physical condition and mental condition.Results(1)The analysis (log rank) showed that the length of hospitalization had no significant statistical differences with relapse(χ2=0.069,P=0.966).(2) The number of hospitalizations (RR=1.074,95%CI=1.002-1.151,P=0.042) and average daily alcohol intake(RR=1.035,95%CI=1.012-1.059,P=0.003) were the risk factors for relapse.ConclusionThe number of hospitalizations and average daily alcohol intake are risk factors for relapse within a year in male inpatients with alcohol-induced psychiatric and behavioral disorders.Prolonged hospital stay has no help to reduce relapse in those people.

OBJECTIVEThis study is to establish the fingerprint and find out the common chromatographic peaks of Inula cappa by HPLC.

METHODThe HPLC analysis was performed on an Agilent Eclipse Plus C18 column (2.1 mm x 150 mm, 1.8 μm) with 0.1% fomic acid aqueous solution-0.1% fomic acid acetonitrile solution as mobile phase at a flow rate of 0.3 · mL(-1) · min(-1); The detective wavelength is 325 nm; The column temperature is 45 °C.

RESULTThe results indicated that 5 of 17 common peaks were identified . The similarity about 10 groups of Inulacappais is over 0.95.

CONCLUSIONThis method is able to be a scientific basis of quality assessment according to its convenient and reliable.