Objective To investigate the functions of Monocyte chemotactic protein-induced protein 1 (MCPIP1) in human breast cancer cell line MDA-MB-231.Methods MDA-MB-231 cells were transfected with GFP-tagged MCPIP1 by Tet-on inducing expression system.Endogenous MCPIP1 was knocked down by stable expressing shRNA.MTT assay was performed to measure the growth of MDA-MB-231 cells after overexpression or knockdown of MCPIP1.FACS method was used to analyze cell cycle in MDA-MB-231 cells.Real-time PCR was used to test the expression of cell cycle-related mRNAs expression and their half-lives.RNA-IP experiment was conducted to detect the mRNA directly enriched by MCPIP1.Luciferase assay was performed to determine whether the mRNA decay was mediated through 3′UTR.Results MCPIP1 overexpression significantly inhibited cell proliferation(P<0.05), while knockdown MCPIP1 promoted cell proliferation with statistical significances (P<0.05).MCPIP1 induced cell cycle arrest in MDA-MB-231 with statistical significance (P<0.01).MCPIP1 overexpression reduced the half-lives of cell cycle mRNAs (CDK2,CDK6,cyclin D1,cyclin E1,respectively) with significance (P<0.01).In addition, cell cycle-related mRNAs were able to be pulled down by GFP-MCPIP1 but not isotype IgG(P<0.05).Compared with control vector, MCPIP1 significant suppressed luciferase activities of all four 3′UTR reporters (P<0.05).Conclusions MCPIP1 functions as a tumor suppressor in human breast cancer cell line MDA-MB-231 through inducing G1 cell cycle arrest.

Zang-fu syndrome differentiation from Huangdi Neijing, laid the theoretical foundation. Hua Tuo'S Zhongzhang Jing, Sun Simiao's Qianjin-Yaofang, Zhang Yuansu Yixue-Qiyuan and other ancient physicians enriched the Zang-fu theory.Zang-fu syndrome differentiation became systematic and widely used in clinical. This paper traced the source of the theory, explored the development of syndrome differentiation theory.

This article discussed the origin and development of the traditional Chinese medicine Shiyan processing, processing history and overview of modern researches. The book of the Summary of information on Chinese herbal medicine processing in all dynasties was consulted which 167 bookson the processing methods of ancient herbal medicine. The content of ancient Shiyan processing was summarized in order to provide standardization for further studies on the optimal processing method of Shiyan, and to ensure the quality of the pieces, and the clinical curative effect.

Objective:To study the effects and mechanisms of acetyl-11-keto-β-boswellic acid ( AKBA) in myocardial ischemic model induced by isoproterenol hydrochloride ( ISO) in rats. Methods:The SD rats were randomly divided into the sham group, model group, AKBA low dose group and AKBA high dose group. Myocardial injury model was induced by subcutaneous injection of ISO (100 mg·kg-1 ) . The change of ST segment in ECG was observed. Creatine kinase ( CK-MB) , cardiac troponin I ( cTnI) , lactate dehydro-genase( LDH) , malondialdehyde( MDA) and superoxide dismutase( SOD) in the blood were detected by ELISA. The change of histo-logical tissue was determined by HE staining, and cell apoptosis was analyzed by TUNEL assay. Results: Serum CK-MB, cTnI and LDH were decreased significantly in AKBA high dose group when compared with those in the model group. Compared with that in the model group, MDA content was lowered and the SOD activity was increased in AKBA high dose group. Furthermore, AKBA high dose group improved the pathologic changes of myocardium. TUNEL assay revealed significant reduction of cardiomyocytes apoptosis in the hearts of the ischemic rats in AKBA high dose group. Conclusion:AKBA has excellent cardioprotective effect on myocardial ischemic induced by ISO and protection of myocardial cells from injury.

Objective To study the changes and the role of MCP -1,IL -8,VEGF,NO,NOS in the T2DM patients with different types of CHD.Methods According to the result of coronary arteriongraphy and clinical symp-toms,and the diagnostic code of T2DM established by Chinese Medical Association diabetology branch in 2007, 126 patients of T2DMwith CHD were chosen and divided into two groups:ACS +T2DM group (A group,74 cases) and SAP +T2DMgroup (B group,52 cases),in addition,50 healthy people were chosen as control group.The levels of MCP -1,IL -8,VEGF were measured by the method of ELISA.The level of NO was measured by the method of nitrate reductase and NOS activity was measured by the method of spectrophotometer.Then,the results were analyzed. Results The levels of MCP -1 and IL -8 in A group and B group were[(115.98 ±39.57)pg/mL,(98.76 ± 31.55)pg/mL],[(131.22 ±42.83)pg/mL,(115.75 ±40.37)pg/mL],which were all higher than those in group C [(75.63 ±23.69)pg/mL,(68.53 ±37.85)pg/mL,t =4.12,2.26,3.78,2.21,all P <0.05)],but the VEGF [(167.87 ±54.98)pg/mL,(128.38 ±36.99)pg/mL)],NO[(46.89 ±12.92)μmol/L,(51.66 ±12.49)μmol/L)]and NOS [(39.04 ±5.19)U /mL,(40.56 ±7.03)U /mL)]were lower than those in C group [(90.21 ± 32.06)pg/mL,(64.05 ±13.58)μmol/L,(47.82 ±5.93)U /mL;t =3.05,3.17,2.43,2.79,2.49,2.25,all P <0.05].The MCP -1,IL -8 levels in A group were higher than those in B group(t =3.13,2.89,all P <0.05),but the level of VEGF and NO were lower than that in B group(t =3.04,2.95,all P <0.05),NOS in A group was lower than that in B group,but there was no statistically significant difference between the two groups(t =1.06,P >0.05). MCP -1 was positively correlated with Il -8,VEGF (r =0.35,0.33,all P <0.01),and it had negative correlation with NO (r =-0.24,P <0.05).Conclusion Inflammatory factor and oxidative stress both participate in the T2DM with different types of CHD,it relates with the degree of CHD.

Objective To summarize the clinical characteristics of primary pigmented nodular adrenocortical disease (PPNAD).Methods Eight patients with PPNAD from 2001 to 2009 in PUMC Hospital were reviewed,their clinical data were collected.Results PPNAD often occurred in adolescents.62.5% of patients with PPNAD were complicated with Carney complex(CNC).In addition to general features of Cushing's syndrome,amenorrhea and growth retardation in stature were very frequent in clinical manifestations of PPNAD.Plasma ACTH was undetectable,circadian rhythm of plasma cortisol was disappeared,glucocorticoid excretion was increased paradoxically during the dexamethasone suppression test in 50% patients with PPNAD.Adrenal imaging from 75% patients revealed normal-sized adrenal glands or suspectable micronodular changes.Adrenal pathologic analysis revealed numerous brown cortical nodules containing lipofuscin pigmentation.Unilateral adrenalectomy may relieve symptoms of Cushing's syndrome,but plasma ACTH and circadian rhythm of plasma cortisol were difficult to recovere.Hypercorticoidism might re-occure after unilateral adrenalectomy.Conclusion PPNAD should be bewared in ACTH independent Cushing's syndrome patients without apparent adrenal mass,and CNC should be screened and followed up.

Objective To investigate of the risk factors for the death of severe fever with thrombocytopenia syndrome (SFTS),so as to set up SFTS critical score and evaluate its role in predicting the prognosis for patients with SFTS.Methods A total of 123 SFTS patients hospitalized in Ji′nan Hospital of Infectious Diseases affiliated to Shandong University from June 2011 to October 2014 were enrolled in this study.The univariate Logistic regression analysis was performed to analysis the risk factor for the death of SFTS.Then the SFTS critical score system was set up accordingly.The prognosis value of SFTS critical score was compared with the rapid emergency medicine score (REMS)and the acute physiology and chronic health evaluation Ⅱ (APACHEⅡ)by using receiver operator characteristic curve (ROC).Results Among all the patients,17 males and 14 females were in death group,and 45 males and 47 females were in survival group.The results of the univariate Logistic regression analyses indicated that the glasgow coma scale (GCS),lactate dehydrogenase,activated partial thromboplastin time,oxygen saturation were risk factors for the death of SFTS,with statistically significant difference (all P <0.05). All of the four parameters of SFTS critical scores in the death group were higher than those in the survival group,with statistically significant difference (all P <0.05 ).The REMS,APACHEⅡ score and SFTS critical score in the death group were significantly higher than those in the survival group (all P <0.01 ). The area under the curve (AUC)of REMS,APACHE Ⅱ scores and SFTS critical score were 0.734, 0.746 and 0.788,respectively.The Youden index of the SFTS critical scores was the highest among all three scores (P <0.01).If 15 .0 was used as the cut off value of SFTS critical score,the specificity and the sensitivity for predicting the death risk for the hospitalized patient were 74.2% and 76.1 %, respectively.Conclusion SFTS critical score,REMS and APACHEⅡ score can all effectively predict the prognosis for SFTS patients,among which,the SFTS critical score is the most convenient and has the best predictive value.

OBJECTIVETo study the chemical constituents of Abelmoschus esculentus.

METHODThe chemical constituents were isolated and purified by chromatography on silica gel and recrystallization. The chemical structures were elucidated on the basis of physicochemical properties and spectral data.

RESULTFourteen compounds were isolated and identified as 6-hydroxy-stigmasta-4-en-3-one(1), 6-hydroxy-stigmasta4,22-dien-3-one(2), stigmasta-5-en-3-ol-7-one(3), stigmasta-5, 22-dien-3-ol-7-one(4), stigmast-5-en-3, 7-diol(5), stigmast-5, 22-dien-3, 7-diol(6), stigmast-4, 22-dien-3, 6-dione(7), stigmasta-4, 22-dien-3-one(8), ergosta-7, 22-dien-3-ol(9), cycloart-25-en-3,24-diol(10), lupeol(11), aurantiamide acetate (12), stigmasterol(13), hexadecanoic acid (14).

CONCLUSIONCompounds 1-12 are obtained from the genus Abelmoschus plant for the first time and also from the Malvaceae for the first time.

Secretory anti-gp96 scFv fragment was expressed in Pichia pastoris to obtain a small molecule antibody that specifically recognizes heat shock protein gp96. The gp96-scFv fragment gene was synthesized and cloned to Pichia pastoris expression plasmid pPICZa-A. Pichia pastoris X33 was electroporated with the linearized recombinant expression vector, and expression of gp96-scFv fragment was induced by methanol. The His-tagged recombinant protein was then purified by affinity chromatography and analyzed with SDS-PAGE and Western blotting assays. The biological activities of recombinant gp96-scFv fragment were determined by Western blotting, Immunofluorescence, ELISA and FACS assays. The gp96-scFv fragment was expressed successfully in Pichia pastoris. About 50 mg of recombinant protein could be purified from 1 liter of the Pichia pastoris culture supernatant. Its molecular weight was about 15 kDa. The gp96-scFv fragment could specifically bind to gp96 protein by Western blotting, immunofluorescence, ELISA and FACS analyses. Pichia pastoris-expressed gp96-scFv fragment specifically recognizes gp96 protein, which could be used for Western blotting, Immunofluorescence, ELISA and FACS analyses.
Blotting, Western ; Chromatography, Affinity ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Membrane Glycoproteins ; immunology ; Pichia ; metabolism ; Plasmids ; Recombinant Proteins ; biosynthesis ; Single-Chain Antibodies ; biosynthesis

Objective To investigate insulin resistance in type 1 diabetes(T1DM)with euglycemic-hyperinsulinemic clamp.Methods Eight cases of newly diagnosed T1DM and 8 cases of newly diagnosed type 2 diabetes(T2DM)were selected.Their insulin sensitivity index(ISI)was evaluated with euglycemic-hyperinsulinemic clamp after 2 week insulin intensive treatment and compared with that of 10 heMthy volunteers(normal control group,NC group).Results Age,BMI,fasting insulin(Fins),fasting C-peptide in the TI DM group were significantly lower than those in the NC group.while waist-to-hip ratio (WHR),systolic blood pressure(SBP),diastolic blood pressure(DBP),TC,TG,LDL-C,HDL-C were not significantly different between the T1DM and NC groups.Age,BMI,WHR,Fins,fasting C-peptide,SBP,TC,TG in the T1DM group were significantly lower than those in the T2DM group.The ISI of the NC,TlDM and T2DM groups were 12.83±1.09,9.95±0.50,3.80±0.20,respectively.There was significant difference among the three groups(P＜0.05).Conclusion The ISI in T1DM Was significantly lower than that in NC group,but higher than that in T2DM.