Objective:To prepare recombinant pCDNA3.1+/Ag85A DNA vaccine encoding tuberculosis Ag85A gene mediated with LipofectamineTM 2000 and to study the effect on T cell subpopulation via oral vaccination.Methods:Recombinant plasmid containing Ag85A using liposome as a vector was constructed and administered to C57BL/6 mice via oral route.Determination of the contents of IFN-r,IL-4 in serum of C57BL/6 mice was performed by double antibody sandwich ELISA.Furthermore,the ability of splenocytes to secret IFN-? and IL-4 was tested by ELISPOT method.When the mice were vaccinated with recombinant eukaryotic expressing vector 5 weeks later,titers of serum antibody against Ag85A were detected by ELISA.The percentage of the CD4+ and CD8+ T cell subsets in the splenocytes was determined by flow cytometry.Results:Lymphocytes obtained from the spleen of liposomal pcDNA3.1+/Ag85A vaccine-immunized mice exhibited lower IFN-? production and higher IL-4 production than those of pcDNA3.1+ vector immunized mice.The number of spleen MNC secreting IFN-? stimulated by Ag85A protein in vitro was significantly lower than that of plasmid vector group.Liposomal pcDNA3.1+/Ag85A vaccine immunized mice elicited higher Ag85A-specific antibodcy titres.The percentage of the CD4+ and CD8+ T cell subsets in the splenocytes was decreased.The subset both were shown the profile of Th2 responses.Conclusion:The oral recombinant plasmid pCDNA3.1+/Ag85A mediated with LipofectamineTM 2000 It shows down-regulation effect on the subsets of CD4+ T cells and CD8+ T cells.The regimen has good immunogenecity and could induce Th2 type humoral response in C57BL/6 mice.The immunization suppresses secretion of IFN-?.It can greatly enhance the titres of Ag85A-specific antibodies.LipofectamineTM 2000 can act as an adjuvant through comparation with negative control group.

Diabetic retinopathy is a progressive vision-threatening complication of diabetes mellitus.The characters of diabetic retinopathy in clude capillary occlusion,microcirculation disturbance and retina neovascularization near ischemic areas of the retina.The exact pathogenic mechanism is still unclear.Recent studies showed that it may be related to vascular inflammation of the retina.Cytokines can cause induction of proinflammatory and adhesion molecules and thereby increase monocyte-endothelial cell adhesion,which is now accepted as the early key event in the development of diabetic retinopathy.The functions of many cytokines such as vascular endothelial growth factor,transforming growth factor-β,basic fibroblast growth factor and ideas for medicine therapeutic to diabetic retinopathy are summarized in this review.

Objective To explore the clinical characteristics of myasthenia gravis(MG)in children.Methods One hundred and six children with MG in Children′s Hospital of Chongqing Medical University from Apr. 2002 to Apr. 2007 were concluded in the study.The data of laboratory examinations,including virus antibody,autoantibody test,immunity function,chest X-ray or chest CT,MRI,ECG and repetitive nerve stimulation, and treatment of cholinesterase inhibitor,glucocorticoids and high-dose immunoglobulin were reviewed.Results Among 106 children,male-to-female ratio was 1 to 1.16,male 49 cases,female 57 cases.The course of disease was 3 days to 11 years.The age of onset of all 106 children with MG varied from 10 months to 14.3 years old and the mean age was (4.74?3.32) years old.The most common age of onset was less than 3 years old(55.7%).There were 41 cases newly diagnosed children and 65 cases received irregular treatment or relapsed after treatment.The ocular muscular symptoms were mostely found at the onset(84.0%);brainstem type was involved at the onset(2.8%),generalized type was involved at the onset(13.2%);58 out of 106 cases(54.7%) had upper respiratory tract infection before the onset with MG; 41 out of 52 cases with MG had abnormal immune function or auto antibody-positive,myocardial enzymes increased in 45 cases increased in varying degrees,thymoma were not found by use of sternum or CT or MRI in 69 cases,repetitive nerve stimulation of 7 out of 11 cases were abnormal; 17 cases (16.0%) of the children received single pyridostigmine treatment,10 cases (9.4%) received co-adrenal hormone and before steroid given IVIG therapy respectively were satisfied with the efficacy,and gravis symptoms were improved;61.3% of the cases had a relapse who underwent irregular treatment or relapse after treatment,before relapse 37 cases (56.9%) received single cholinesterase inhibitor treatment, 28 cases (43.1%)for co-adrenal hormone, after adrenocortical hormone and pyridostigmine treatment.The prognosis of 72 patients(67.9%) with MG was usually good; the causes of relapse resulted from infection (30 cases, 46.2%), irregular drug use (22 cases,33.8%), and drug withdrawal and infection (13 cases, 20.0%).Conclusions As early as age of onset,the disease easily repeated,to be early co-adrenal hormone therapy group is the children′s clinical features of MG.

The property theory of Chinese herbal medicine (CHM) is regarded as the core and basic of Chinese medical theory, however, the underlying mechanism of the properties in CHMs remains unclear, which impedes a barrier for the modernization of Chinese herbal medicine. The properties of CHM are often categorized into cold and heat according to the theory of Chinese medicine, which are essential to guide the clinical application of CHMs. There is an urgent demand to build a cold/heat property classification model to facilitate the property theory of Chinese herbal medicine, as well as to clarify the controversial properties of some herbs. Based on previous studies on the cold/heat properties of CHM, in this paper, we described a novel strategy on building a cold/heat property classification model based on herbal bio-effect. The interdisciplinary cooperation of systems biology, pharmacological network, and pattern recognition technique might lighten the study on cold/heat property theory, provide a scientific model for determination the cold/heat property of herbal medicines, and a new strategy for expanding the Chinese herbal medicine resources as well.
Drugs, Chinese Herbal ; chemistry ; therapeutic use ; Herbal Medicine ; Humans ; Phytotherapy ; Plants, Medicinal ; chemistry

Objective:To prepare a colloid gold kit for the qualitative detection of the dopes met-amphetamine and morphine by one-step chromatography immunoassay in human samples based on the principle of the highly specific immunochemical reactions between the antigens and their antibodies which were used for the analysis of specific compounds in human urine samples at ideal cut-off concentrations.Methods:The monoclonal antibodies(McAb) of either mouse anti-morphine or mouse met-amphetamine were prepared by hybridomas and the limiting dilution respectively. The titers of McAb were tested by ELISA.The kit was assembled and the specificity,sensitivity,and stability were investigated.In addition,these characters were compared with those of issued Pan Probe single test strip.Results:①The titer of the first McAb against morphine was 1∶3.2?103, as for another, 1∶1.6?103. ②The colloid gold kit was found with following cut-off for the dope concentrations,morphine at 300 ng/ml and met-amphetamine at 1 000 ng/ml. ③The colloid gold kit showed fine specificity without cross-reaction with routine medicines for cold treatment or with the solf-drinks of cokocola or with tobacco for the individuals who did not take the dopes the 7 days.④The colloid gold kit was even more sensitive than the issued strip,because urine sample of the junkies were still positive 4-5 days post-intake.Conclusion:The colloid gold kit for indentifying the drug-users is a rapid,specific, sensitive immunoassay suitable for the simultaneous, qualitative detection of drug abuse.

Objective To observe inhibitory effect of recombinant human endostatin on the proliferation of vascular tumor endothelial cells and to investigate its possible mechanism.Methods Hemangioma endothelial cells were cultured in vitro and different concentrations of endostatin on hemangioma endothelial cell proliferation inhibition were detected by MTT method.Effect of recombinant human endostatin on endothelial cell cycle was detected by flow cytometry.The expression of VEGF, KDR mRNA in hemangioma endothelial cell were detected by Real-time RT PCR. Results Recombinant human endostatin concentrations in 24 h, 48 h and 72 h after three period during which the hemangioma endothelial cells inhibited significantly( P <0.01 ) , and there was a clear dose dependence, IC50 was 355 μg/mL.Recombinant human vascular endostatin ( 250 μg/mL ) intervented for 24 hours, the proportion of cells in G0/G1 phase(94.23 ±1.66)%, compared with control group (90.63 ±1.14)%, had significantly difference (P<0.05).Compared with the control group, the difference of the expression of vascular endothelial growth factor (VEGF) was statistically significant (P<0.05) as well as Flk-1 (P <0.05).Conclusion Recombinant human endostatin does not only have inhibitory effect of hemangioma endothelial cell cycle, but also can inhibit the expression of VEGF and FLK-1.

Objective To systematically evaluate the efficacy of alendronate on the primary and secondary preventions of osteoporotic fractures in postmenopausal women.Methods Literatures and related references were searched from MEDLINE (from 1966 to April 2011),Embase (from 1966to April 2011),Cochrane Library (the second Issue,2011),CBM disc,CNKI,WANFANG and VIP database by Cochrane systematical evaluation.All related randomized controlled trials were collected,and Meta analysis was performed by RevMan 5.0 soft ware provided by Cochrane collaboration.Results Totally 11 randomized controlled trials were evaluated.Meta-analysis showed that 10 mg oral alendronate daily for secondary prevention could reduce the incidence of spine,hip,wrist and non vertebra fractures (RR 0.57,95％ CI:0.49-0.67),but for the primary prevention,alendronate could not reduce the fracture incidence except for spine fracture (RR=0.55,95％CI:0.38-0.80).Adverse events associated with alendronate were not increased,and the sensitivity analysis results were not changed (RR =0.95,95％ CI:0.83-1.09).Conclusions Alendronate can reduce osteoporotic fractures in postmenopausal women for the secondary prevention,but the effect of alendronate on osteoporotic fractures for the primary prevention needs further study.

Saliva and samples containing saliva were regarded as a kind of potential biological trace evidencefor individual identification in forensic science practice. In this paper, the authors have studied theDNA analysis of saliva and saliva containing samples. The results demonstrated that DNA can be extracted from saliva and saliva-containing samples. RFLPs analysis by DNA fingerprinting and PCRbased typing can be performed according to the amount of DNA extracted from these samples.

Objective To investigate the glucocorticoid treatment of nephrotic syndrome curative effect and the effect of psychological intervention on the patients. Methods From June 2015 to May 2016 in our hospital treated 110 cases of patients with nephrotic syndrome, patients were randomly divided into two groups, control group patients with routine treatment and nursing measures, observation group of patients on the basis of conventional treatment nursing glucocorticoid treatment and nursing intervention measures for nursing, completion of treatment, compared two groups of patients with therapeutic effect and recurrence. Results In terms of treatment, observation group of patients treatment effectiveness 94.55％, control group patients treatment effective rate was 81.82％, the observation group of patients treatment effectiveness is significantly higher than patients with the control group (P<0.05); In terms of relapse, observation group after treatment in patients with recurrence rate was 3.64％, the control group after treatment in patients with recurrence rate 9.09％, observation group of patients with recurrence rate was significantly lower than patients with the control group (P<0.05). Conclusion By glucocorticoid treatment of nephrotic syndrome curative effect and psychological intervention on the patients, can significantly improve the effect of treatment of nephrotic syndrome patients, reduce the occurrence of patients relapse, help patients recover better, worthy of popularization and application in clinic.

Objective To investigate the food intake of one meal of the hotel customers and evaluate it rationality ,then provide dietary suggestions .Methods Six hundreds hotel customers were investigated in 2 four‐star hotel and 2 five‐star hotel of Nanjing , who were selected with a stratified sampling method .Their 5 days diet were investigated and then the dietary intake ,the percentage of energy from protein ,fat and carbohydrate and the percentage of protein resource were calculated based on the food composition table .Results It was shown that the daily energy intake of one meal was appropriate among the surveyed hotel customers ,and the energy proportion from 3 major nutrients were not reasonable ,and the energy proportion from fat were higher than other 2 major nutrients ;the proportion from the source of good proteins was reasonable ,but the intake of protein was too much;the customers fell short on their daily serving of grains ,eggs ,beans and milk products and the intake of cooking oil and salt was excessive in diets . Their intake of fat ,protein ,retinol equivalent ,niacin ,vitamin E ,sodium and iron were too much .Instead ,they ate less ascorbic acid;only the intake of energy ,riboflavin and thiamine were appropriate .Conclusion The one meal dietary structure of hotel customers is not appropriate ,while nutrition collocation is not reasonable .It is necessary to promote nutrition and health education and some intervention measures must be taken .