Objective:To evaluate the antitumor activity of recombinant SEA for therapy of B16 melanoma established in C57BL/6 mice. Methods:C57BL/6 mice with melanoma were treated with the purified rSEA. The tumors were isolated and weighted. Results:Tumor growth was apparently inhibited by rSEA at high, middle, and low doses intraperitone-ally, whose inhibition ratio were 79.3% , 75.6 % and 73. 8% respectively. rSEA treatment in situ could inhibit tumor growth more effectively(90.6% ). Further study showed that numerous CD8+ and CD4+ T cell were infiltrated in tumor tissues, which were consistent with tumor growth inhibition induced by rSEA. Conclusions: rSEA could inhibit tumor growth effectively, especially the treatment in situ. This study paves the way for tumor immunotherapy with targeted SEA.

Objective:Helicobacter pylori is one of the human pathogens causing gastric ulcers and cancers.A key virulence factor of H.pylori is the Cag pathogenicity island,which encodes a type IV secretion system.HP0525 is an essential component of the Cag system and acts as an inner membrane associated ATPase.To construct recombinant plasmid containing hp0525 gene of Helicobacter pylori(H.pylori)NCTC 11637,and analysis of sequence nucleic acid,express it in E.coli BL21 and study the influence of the HP0525 protein on proliferation of SGC-7901 cells.Methods:H.pylor hp0525 gene was amplified from the genome DNA by PCR,then operated T-A cloning and sequenced.The hp0525gene fragment was inserted directionally into vector pMD18-T to construct recombinant clones of hp0525 and was sequenced.The recombinant plasmid was transformed into E.coli BL21for expressing under induction of IPTG.Purify the expressed protein by Ni2+-NTA column chromatography.Expressed product was analyzed by Western blot and MALDI-TOF.Added the purified protein into SGC-7901 cells,the effect of cell proliferation in SGC-7901 cells induced by the recombinant protein was observed by MTT.Results:A 993 base pairs long hp0525 gene,which encodes a product of 330 amino acid,was obtained using PCR method and was cloned into pMD18-T vector successfully.The sequence analysis for hp0525 showed that it shares 97%~99% homology with other strains in Gene bank.SDS-PAGE showed a protein band with a relative molecular weight of 36 000,which was consistent with the expectation.The expressed product reached a purity of 97% after Ni2+-NTA column chromatography.The protein after dialyzed and annealed,was co-cultured with SGC-7901 cells,The protien of different concentration co-cultured with SGC-7901 cells for different times,found that the protein in low concentration stimulates proliferation of cells,to achieve some concentration,it inhibits proliferation of cells along with multiplication of the concentration of the protein;The protein inhibits proliferation of cells relay on the extension of time and concentration.Conclusion:It is indicated that the correct hp0525 gene was obtained and expressed in E.coli BL21.High purified protein was obtained by Ni2+-NTA column chromatography,the protein can inhibits proliferation of SGC-7901cells,and T-ATPase activity which posed a basis for further researching on its biological function.

Objective: To optimize the extraction process of volatile oil from Fructus Litseae.Methods: Supercritical CO2 extraction (SFE-CO2) was used to extract volatile oil from Fructus Litseae, and the extraction process was optimized by a central composite design-response surface method (CCD-RSM).The kind and content of volatile oil extracted by SFE-CO2 and steam distillation (SD) were detected by gas chromatography-mass spectrometer (GC-MS).Results: The highest extraction rate of the total effective constituents was 66.26% when the optimum conditions were as follows: the extraction temperature was 44 ℃, the extraction pressure was 26 MPa, entrainer ethanol was 11 ml and the extraction time was 40 min.It was revealed that neral, geranial and limonene were the major constituents.Compared with that of SD, the kind of volatile oil extracted by SFE-CO2 was fewer while the content of citral extracted by SFE-CO2 was higher.Conclusion: The extraction process optimized by CCD-RSM provides reference for the extraction method and quality control of volatile oil from Fructus Litseae.

Objective To compare the survival rates from two conservative strategies in the treatment of malignant pleural mesothelioma (pemetrexed plus cisplatin or pemetrexed plus carboplatin ).Methods Seventeen cases diagnosed of malignant pleural mesothelioma at clinical stages of or over Butchart Ⅲ in our hospital during 2005 -2009 were treated with pemetrexed plus cisplatin (10 cases,Group A)or pemetrexed plus carboplatin (7 cases,Group B ).The difference in the survival rates between these two groups was compared.Results The early survival rate (0 -3 months)in pemetrexed plus cisplatin (group A)-treated group was more than that of pemetrexed plus carboplatin group (group B),but the difference was not statistically significant (x2 =0,1.52,1.52,respectively,P ＞ 0.05 ).For the medium-term survival rate (4 - 9 months),group A was better than group B,and the difference was statistically significant( x2 =5.21,4.41,4.41,4.50,4.50,4.55,respectively,P ＜ 0.05 ).The two groups produced comparable long-term survival rate s (10 -12months) ( x2 =1.31,0.09,0.09,respectively,P ＞ 0.05 ).Conclusion To increase the medium-term survival rate (4 -9 months)of patients with MPM,we prefer to using pemetrexed plus cisplatin regime.For the patients with more severe side effects which can be relieved by folic acid and VitB12,this regime is also recommended.Otherwise,for the patients with unrelievable side effects,especially for those with poor physical condition or a short survival expectation,pemetrexed plus carboplatin is suggested.

Objective To explore the direct association between metabolic syndrome (MS) and gamma glutamyltransferase (GGT) or alanine aminotransferase (ALT) in Beijing men and women.Methods We investigated the medical check-up data from the 14,204 physical examination population (male 9524,female 4680) of year 2008.A cross-sectional study was carried out to explore the direct association between MS and GGT or ALT.MS was defined by the Chinese Diabetes Society (2004).Chisquare test was performed to analyze the categorical variables.The receiver operating characteristic curve (ROC) was plotted to obtain area under curve (AUC) and the cutoff point of GGT and ALT.Results The prevalence of MS was significantly higher in subjects with the highest quartile of GGT or ALT than the subjects with the lowest quartile of GGT or ALT.For male,the prevalence of MS is 7.57％ in the lowest GGT quartile and 29.04％ in the highest GGT quartile.For female,the prevalence of MS is 1.28％ in the lowest GGT quartile and 17.86％ in the highest GGT quartile.For male,the prevalence of MS is 9.27％ in the lowest ALT quartile and 21.88％ in the highest ALT quartile.For female,the prevalence of MS is 3.52％ in the lowest ALT quartile and 14.70％ in the highest ALT quartile (P ＜0.01).The optimal cutoff points of GGT to diagnose MS were 31 U/L in men and 18 U/L in women.The optimal cutoff points of ALT to diagnose MS were 23 U/L in men and 19 U/L in women.Conclusions The prevalence of MS increases with the increase in serum levels of GGT or ALT even through the normal range of GGT or ALT in Chinese men and women.

Objective To explore the application of CO2 as an insufflation agent in patients under-going colonoscopy by comparing carbon dioxide(CO2 )and air.Methods Consecutive 681 patients who un-derwent conventional colonoscopy from January to May in 2014 at endoscopy center of our hospital were in-cluded.The patients were randomly divided into the air group and CO2 group.The gas flow,capacity,oper-ation time,PaCO2 and abdominal pain during and 20 min,40 min,1 h,2 h,12 h,24 h after colonoscopy were recorded.The safety and advantages of CO2 in the colonoscopy were analyzed.Results The intubation time of CO2 group was significantly shorter than that of air group (P <0.05),but there was no significant difference in endoscopy withdrawal time between the two groups (P >0.05).There was also no difference in air flow and the total volume between the two groups (P >0.05).PaCO2 of two groups were within the nor-mal range.PaCO2 during examination was significantly higher than that before and 20 minutes after examina-tion (P <0.05),but the PaCO2 between the two groups showed no significant difference (P >0.05).The abdominal pain scores during and 20 min,40 min,1 h,2 h after the examination of air group were signifi-cantly higher than those of CO2 group (P <0.05),but they were not 12h and 24h after the examination (P>0.05).Conclusion CO2 is safe and could reduce abdominal discomfort after colonoscopy,which is suit-able for clinical application.

BACKGROUND:Transgenosis of basic fibroblast growth factor (bFGF) gene has been successfully performed into the muscle satellite cells of rat extraocular muscles in the previous study of the research group, proving that bFGF could express in the myoblasts of extraocular muscles, also promote cell proliferation and differentiation.OBJECTIVE: To further investigate the methods for regulating the expression of the bFGF in myoblasts following transfection. METHODS: Target gene bFGF was connected with inducing expression vector pcDNA4/T0/myc-His?A, followed by masculine clone sequencing identified by colony PCR and enzyme digestion, EcoR I and Hind III restriction enzyme digestion, as well as Xho I single enzyme verification. C2C12 myoblasts antibiotics sensitivity was screened and finally defined. By use of lipofection transfection technology, cell lines where C2C12 stably expressed pcDNA6/TR were estabolishd and then identified by Western blot. The pcDNA4/TO/myc-His?A-bFGF was transfected into pcDNA6/TR- C2C12 cells. The bFGF expression and secretion in C2C12 cells following tetracycline-induced pcDNA4/TO/myc-His?A-bFGF transfection were determined by immunofluorescence and Western blot, the controls were established.RESULTS AND CONCLUSION: ① The conjunction between the bFGF and inducing expression vector pcDNA4/TO/myc-His?A was proved successfully by sequencing comparison, double digestion and single digestion. ②The minimal lethal concentration of blasticidin to C2C12 cells was 10 mg/L, while that of zeocin was 750 mg/L. ③ The pcDNA6/TR-C2C12 cell lines were established correctly. ④ The myoblasts treated by tetracycline and transfected with pcDNA4/TO/myc-His?A-bFGF were positive for gene expression, those untreated exhibited a negativity; bFGF protein could be produced in myoblasts treated by tetracycline and transfected with pcDNA4/TO/myc-His?A-bFGF, the production reached a peak at 24 hours, while those untreated can not produce bFGF protein. Results suggest that the bFGF expression in the myoblasts can be controlled by tetracycline inhibition and regulatory systems.

OBJECTIVE:To investigate the protective effect of Tibetan medicine Compound Duoxuekang capsules on the acute lung injury in mice induced by lipopolysaccharide. METHODS:60 KM mice were randomly divided into blank group,model group,dexamethasone group (positive control,1 mg/kg) and Compound Duoxuekang high-dose,medium-dose,low-dose groups (3.6,1.8,0.9 g/kg,calculated by crude drug),10 in each group. Intragastrically administrated,mice in blank group and model group intragastrically administrated equal volume of normal saline,once a day. After 7 d of administration,except for blank group, mice in other groups intraperitoneally injected lipopolysaccharide to induce acute lung injury. After 6 h modeling,pathological changes in the lung tissue was observed,lung tissue water content was measured,superoxide dismutase(SOD),glutathione peroxi-dase(GSH-Px),malondialdehyde(MDA)levels in lung tissue and IL-6,TNF-α levels in serum were detected. RESULTS:Com-pared with blank group,mice in model group showed pathological changes in congestion and edema,inflammatory cell infiltra-tion,obvious widened alveolar septum and alveolar wall;water content in lung tissue and IL-6,TNF-αlevels in serum were signifi-cantly increased (P<0.01);SOD,GSH-Px levels in lung tissue were significantly decreased (P<0.01). Compared with model group,injury degree of lung tissue reduced to varying degrees in each treatment group,except for the water content,MDA level in lung tissue and TNF-α level in serum in Compound Duoxuekang capsules low-dose group;the above-mentioned indexes in other groups were significantly improved (P<0.05 or P<0.01). CONCLUSIONS:Compound Duoxuekang capsules can obviously re-duce mice's oxidative stress and inflammatory response,and has certain protective effect on acute lung injury in mice.

The effects of Traditional Chinese medicine (TCM) nursing operation in cervical spondylitis have been generally recognized. This paper reviewed the applications and improvements of the major three TCM nursing operations, cupping, Gua-Sha and acupressure. The future developments in moxibustion, auricular point sticking, fumigation and washing therapy and moisten compress therapy were also prospected.

Objective:To analyze the value of pepsinⅠ( PGⅠ) , pepsinⅡ( PGⅡ) , gastrin-17 ( G-17 ) diagnostic and Helicobacter pylori( Hp IgG) antibody in screening chronic atrophic gastritis and gastric cancer.Methods:90 patients with an upset stomach in our hospital from May 2014 to May 2015 were selected for the study, according to the pathological diagnosis which were divided into normal control group( including chronic non-atrophic gastritis) ,chronic atrophic gastritis group and gastric cancer,30 cases in each group.The level of PGⅠ, PGⅡ, G-17 and Hp IgG antibody positive rate in the three groups of patients were compared.Results:The level of PGⅠ,PGⅡin gastric cancer patients were lower than the control group and chronic atrophic gastritis group.And the index of chronic atrophic gastritis patients were lower than control group.The level of G-17 in gastric cancer group was higher than chronic atrophic gastritis group and the control group, and chronic atrophic gastritis group and the control group had no significant difference.The Hp IgG antibody positive rate in gastric cancer group was significantly higher than other two grups.The level of PGⅠand PGⅡin Hp-infected patients were lower than uninfected Hp patients,and the level of G-17 was higher than uninfected Hp patients.The level of PGⅠ, PGⅡ were significantly negatively correlated with age, pathological stage and metastasis, and positive related with the degree of differentiation;but the level of G-17 and Hp IgG antibody rate were positive related with age,pathological stage and metastasis, and negatively correlated with the degree of differentiation.Conclusion: PGⅠ, PGⅡ and Hp IgG antibody screening have a good diagnostic value in chronic atrophic gastritis and gastric cancer,and better diagnostic value of gastric cancer,G-17 diagnostic value of gastric cancer is much better than the chronic atrophic gastritis;and the level of PGⅠ,PGⅡ,G-17 and IgG antibody positive rate are closely related to the clinicopathological features of gastric cancer patients.