Objective To establish determination method for total polyphenols in Guhong Injection, and provide reference for internal quality control of Guhong Injection.[Methods]Using the Folin-Ciocalteu colorimetric method, according to the consistent degree of the maximum absorption wavelengths of hydroxysafflor yel ow A and kaempferol relative to Guhong Injection, we selected the appropriate reference and color reaction condition to content determination.[Rsults]With hydroxysafflor yel ow A as the standard, the content of total polyphenols in 10 batches of samples were 9.94, 9.55, 9.75, 9.67, 9.84, 10.03, 9.81, 9.52, 9.88, 10.09mg·mL-1. The average recovery of total polyphenols was 98.11% and RSD was 1.68%(n=6). [Conclusion]The established Folin-Ciocalteu method is simple, accurate, sensitive, and is reliable for detecting total polyphenols in Guhong Injection.

AIM: To decide the effect that selected siRNA degrades mRNA of IL-1? specifically and suppression of its expression after connected with target site with homology complementary sequence. METHODS: Synthesized DNA expression box aimed directly at target site through PCR reaction in vivo was purified, and transfected into lymphocytes stimulated by LPS. siRNA was transcribed by cellular endogenous RNA polymerase Ⅲ and then evoke the degradation of target mRNA. After 48 hours of transfection, the cell culture supernatant was collected and the concentration of IL-1? was assayed using ELISA. RESULTS: Compared with blank-control and negative-control, selected sequence decreased the expression of IL-1?. Rate of the suppression was about 15%. CONCLUSION: RNAi technology produces specific interference effect in mouse spleen lymphocytes in original culture and inhibits the excretion of IL-1?.

AIM: To explore the effect of nuclear factor-?B (NF-?B) on the anti-apoptosis induced by brain ischemia preconditioning (IP). METHODS: Temporary middle cerebral artery occlusion for 20 min followed three days reperfusion before 6 hours middle cerebral artery occlusion (MCAO) trancranially was used as preconditioning in Wistar rats. The protective role was evaluated by analyzing the infarct volume. The status of neuronal apoptosis was observed by TUNEL. The expression of NF?B p65 protein, the assay of SOD activity and MDA concentration were analyzed by using the methods of immunohistochemistry and cytochemistry. RESULTS: Compared to the control group, 20 min ischemic preconditioning, which did not produce neuronal damage obviously, reduced the infarct volume significantly after MCAO 6 h and obviously decreased the number of neural cell apoptosis in penumbra (P

Objective To investigate the effects of type 2 diabetes on learning and memory of APP/PS1/Tau triple transgenic (3 × Tg) mice of Alzheimer’s disease, and the protective mechanism of liraglutide (LIR) thereof. Methods One month old C57BL/6 mice were set to be control group (WT). One month old 3×Tg mice were divided into control group (Tg), liraglutide group (Tg+LIR), type 2 diabetes group (Tg+T2DM) and liraglutide treatment group (Tg+T2DM+LIR). The model of T2DM was established by feeding the high fat and sugar fodder, and then injecting streptozotocin (STZ) in mice, making sure the fasting blood glucose was more than 7 mmol/L. Then the subcutaneous injection of LIR was administered for 2 months. The values of body weight and fasting blood glucose were detected at age of 5-month. Morris water maze was applied to evaluate the spatial learning and memory ability. Western blotting assay was used to measure the levels of phosphorylated Tau, neurofilament (NFs) and insulin receptor substrates. ELISA was used to detect the human Aβ42 to evaluate the effect of LIR on-amyloid. Results LIR can reduce body weight and blood glucose, can alleviate spatial learning and memory damaging caused by T2DM, and also can improve phosphorylated Tau levels, NFs and insulin receptor substrates caused by T2DM, and finally can reduce the deposition ofβ-amyloid of 3 × Tg mice. Conclusion T2DM can aggravate symptoms of AD in 3×Tg mice, and LIR has a protective effect on it.

Objective To find out the expression relation between TP53 and NOTCH1,and to explore their effects in head and neck squamous cell carcinoma.Methods Obtained the differentially expressed genes data of head and neck squamous cell carcinoma from 279 samples in TCGA database.Analyzed the co-expression relation between TP53 and NOTCH1 through Pearson and Spearman method.Cbioportal was used to analyze their co-expressed genes.Establish the co-expression network of TP53 and NOTCH1 with String database.The pathway and function of co-expression network was identified through KEGG and DAVID database respectively.Results Among the 279 samples,TP53 and NOTCH1 was co-expressed in head and neck squamous cell carcinoma.(Pearson score =0.45;Spearman score =0.41) There were 182 interaction pairs of TP53 and NOTCH1 related co-expressed gene according to the String database.(Pearson and Spearman score ＞ 0.3)These genes were enriched in some pathways such as T cell receptor signaling pathway,cell cycle,cell adhesion molecules and so on.These genes were enriched in some tumor related function including immune response,regulation of transposition,regulation of apoptotic process,cell cycle,regulation of GTPase activity and so on.Conclusion TP53 and NOTCH1 was co-expressed.Through establishing co-expressed network of TP53 and NOTCH1 and bioinformatics analysis,their function and signaling pathway were explored.The data generated from this study could provide a new reference in mechanism research of head and neck squamous cell carcinoma.

Objective:To confirm the potential of growth-related gene productβ(GROβ) as a biomarker for colorectal cancer. Methods:Serum GROβlevels in 123 subjects with colorectal cancer, 88 healthy controls, and 125 subjects with other diseases were measured using enzyme-linked immunosorbent assay. Serum levels of carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA19-9) in all subjects were measured using immunoluminometric assay. Statistical analyses were conducted to determine the associa-tions between serum GROβlevels and clinical parameters for colorectal cancer. The receiver operating characteristic (ROC) curves of GROβ, CEA, and CA19-9 were analyzed. Results:The serum GROβlevels were higher in patients with colorectal cancer (median=96.15 pg/mL) than in the healthy controls (median=43.28 pg/mL, P<0.01) and in patients with other diseases (median=57.30 pg/mL, P<0.01). The serum GROβlevels in patients with colorectal cancer were positively correlated with the tumor-node-metastasis staging (P<0.01) and depth of infiltration (P<0.05), but not with the histological grade, tumor embolus, lymph node metastasis, gross pathologic tu-mor type, or gender of the patients. The sensitivity and specificity of the assay for serum GROβwere 56.1%(69/123) and 95.31%(203/213), respectively. The diagnostic sensitivity was 22.2%(4/18) for stage I and 66.7%(26/39) for stage II when the data of GROβwere combined with the data of CEA and CA19-9. The ROC curve constructed with the data of GROβ(0.834) was larger than that construct-ed with the data of CEA (0.739) or CA19-9 (0.676) for discriminating colorectal cancer from the matched controls. Conclusion:These preliminary results indicated that the serum GROβlevel could be a useful biomarker for colorectal cancer diagnoses.

Objective:To explore the proportions of Th17 cells in the peripheral blood and levels of IL-17,IL-23 in the serum of patients with Graves′disease ( GD ) and their clinical significance.Methods: We studied 29 patients with GD ( GD group ) , and reevaluated the GD group after therapy ( euthyroid GD group ).29 gender-and age-matched volunteers were selected as the normal control ( NC group).The proportions of Th17 cells were investigated by flow cytometry.The levels of IL-23,IL-17 in the serums were measured by ELISA.The levels of FT3,FT4,TSH were determined by ECLIA and the levels of TrAb were tested by RRA.Results:There were no significant difference among 3 study groups in sex and age match ( F=0.0075 , P>0.05;χ2=0.4213 , P>0.05 ).The proportions of Th17 cells and the levels of IL-17 , IL-23 were increased in the GD and euthyroid GD patients compared with the control group (respectively,P<0.05).The proportions of Th17 cells and the concentrations of IL-23 in euthyroid GD group were significantly lower than those of GD group ( respectively , P<0.05 ) , but there were no significance in the concentrations of IL-17 between euthyroid GD group and GD group(P>0.05).Correlation analysis revealed that the proportions of Th17 cells ,and the levels of IL-17,IL-23 were positively correlated with the levels of FT3,FT4,TrAb(r=0.588 2,0.337 2,0.371 0;0.549 6,0.287 5,0.342 7;0.361 0,0.420 8, 0.330 8;P<0.05 ,for all parameters ) ,and were negatively correlated with the levels of TSH ( r=-0.319 7 ,-0.472 8 ,-0.428 2;P<0.05,for all parameters).Conclusion:Th17 cells and their related cytokines IL-17,IL-23 are highly expressed in the serum of patients with GD.Th17 cells and their relative cytokines have certain relevance with 4 thyroid function parameters of the patients with GD , which can be used as biological markers for GD.

Aim To detect the expression of hippocam-pus NGF gene in the mouse model of Parkinson 's dis-ease. Methods By intraperitoneal injection of 1-meth-yl-4-phenyl-1,2, 3, 6-tetrahydropyridine ( MPTP), a mice model of Parkinson ’ s disease was established. The success of PD model was identified by detecting the expression of mesencephalic tyrosine hydroxylase ( TH) with Western blot and immunofluorescence. The movement and cognitive ability of mice were evaluated by foot print analysis and step-through passive avoid-ance test. The expression of NGF gene in hippocampus of mice was detected with RT-PCR and in situ hybrid-ization. Results Compared with the control group, the levels of TH and NGF were reduced in the experi-mental group and the behavioral indexes were deflected significantly. Conclusion NGF may play an impor-tant role in the occurrence and development of the PD cognitive disorder.

Objective:To explore the role of cytokines in the pathogenesis of Graves′disease(GD),by detecting the levels of IFN-γ,IL-6,IL-17 and TGF-β1 in GD patients who were newly diagnosed.Methods:A total of 23 patients with new onset GD and 23 gender-and age-matched healthy controls were examined.The levels of serum IFN-γ, IL-6, IL-17 and TGF-β1 were measured by ELISA,FT3,FT4 and TSH levels were determined by ECL IA;TrAb levels were tested by RRA.Results: There were no significant difference among GD and NC group in sex and age match ( t=0.334 8 ,P>0.05;χ2=0.410 7 ,P>0.05 ).The levels of serum IFN-γ,IL-6,IL-17 and TGF-β1 in the GD group were significantly higher than the control group ( P<0.05 ) .Correlation analysis revealed that IFN-γ,IL-6,IL-17 and TGF-β1 were positively correlated with FT3,FT4(r=0.324 6,0.453 2,0.431 0,0.463 8;0.413 2,0.441 5, 0.436 2,0.467 1;P<0.05 ).Conclusion: IFN-γ, IL-6, IL-17 and TGF-β1 are highly expressed in the newly diagnosed GD patients.They play an important role in the pathogenesis of GD ,and provide helpful evaluation indices of immune dysfunction to Graves disease.

Objective To investigate the gene mutations of human immunodeficiency virus (HIV)‐1 drug resistance among anti‐retrovirus (ARV ) treated‐naive men who have sex with men (MSM ) in Shanghai to provide evidence‐based data for optimized treatment .Methods All 669 treatment‐nave cases of HIV‐1 infection identified among MSM in 2013 were recruited and their plasma was collected .RNA was extracted and amplified by nest reverse transcription‐polymerase chain reaction ,and DNA was sequenced and then phylogenetically analyzed .Finally ,subtypes were identified and drug resistance was analyzed in comparison with International HIV Drug Resistance Database .Results The pol gene fragments of 645 cases were obtained .Primary drug‐resistance rate was 2 .48% (16/645) ,including mutations conferring resistance to protease inhibitor (PI) (0 .31% ,2/645) ,nucleoside reverse transcriptase inhibitors (NRTI) (0 .16% ,1/645) ,non‐nucleoside reverse transcriptase inhibitors (NNRTI) (1 .70% ,11/645) and both NRTI and NNRTI (0 .31% ,2/645) ,respectively .Mutations conferring resistance to CRF01_AE were 12 cases (2 .99% ) ,while mutations conferring resistance to CRF07_BC and CRF_01B were 0 .61%(1/163) and 4 .65% (2/43 ) including 1 case of CRF52_01B and unidentified CRF_01B , respectively . Resistance to NNRTI in B subtype were 2 .70% (1/37) .Conclusion The prevalence of HIV‐1 drug resistance‐associated mutations among MSM in Shanghai ,2013 is still low ,but resistance to NNRTI is relatively high .CRF01_AE is the major subtype of drug resistance .It is necessary to strengthen the HIV drug resistance surveillance in MSM group in Shanghai .