Membrane type-1 matrix metalloproteinase (MT1-MMP or MMP14) plays the pivotal role in tumor development and metastasis, so it is a promising drug target in malignancy. To acquire MT1-MMP specific binding peptides, we first analyzed MMPs sequences to find the divergent and specific sequence of MT1-MMP by bioinformatics approach, then set the specific sequence as the sense peptide target and designed antisense peptide library. Finally, by means of molecular docking, molecular dynamics simulation and in vitro cell assays, we screened the antisense peptide library against MT1-MMP and further studied the obtained specific peptides. Here, we identified the divergent and specific sequence of AYIREGHE (Named MT1-loop) located in MT1-MMP loop by multiple sequence alignment and established the antisense peptides library with capacity of 1 536 sequences. After two rounds of virtual screening, we obtained five antisense peptides with Rerankscores in the top for further screening. They all interacted with MT1-MMP, and docked well at the active site composed of MT1-loop sequence. Analysis of the affinities of these five antisense peptides to other MMPs (MMP1-3, MMP7-13, MMP14 HPX, MMP16) revealed that the peptide FVTFPYIR was more specific to MT1-MMP. Molecular dynamics simulation showed that the peptide FVTFPYIR might affect the stability of MT1-MMP and thus have effects on its activities. Meanwhile, the peptide FVTFPYIR could specifically inhibit the growth of MG63 and MDA-MB-231 tumor cells both of which expressed MT1-MMP. The work provides a new insight and way for the development of antitumor lead peptides targeting MT1-MMP.
Amino Acid Sequence ; Humans ; Matrix Metalloproteinase 14 ; chemistry ; Molecular Dynamics Simulation ; Neoplasms ; Peptide Library ; Peptides ; chemistry

A label-free electrochemical immunosensor using hollow structure nanomaterials based on its ordered porous and big surface area was designed. Au nanocage, with good conductivity, catalysis, and biocompatibility, was prepared and modified on the surface of glassy carbon electrode with graphene to immobilize antibody of microcystin directly. In the absence of microcystin, biosensor can obtain high current response signal of electrochemical probe ( [ Fe( CN) 6 ] 3-/4-. When microcystin was combined with its antibody specifically, the charge density and mass transfer resistance on the surface of electrode increased, resulting in a decrease of the corresponding peak current of [ Fe ( CN ) 6 ] 3-/4-. This change was in proportion to the concentration of microcystin indirectly. Experiment conditions such as cultivation time of antigen and concentration of antibody were optimized. The results showed wide linear range of 0. 05 μg/L-1. 0 mg/L and the detection limit of 0. 017 ng/mL. This sensor has good stability and simple production procedure. This sensor provides a new and simple means for the ultrasensitive determination of microcystins in real water samples.

Aim To explore the anti-proliferation effects of curcumin trinicotinate ( CurTn ) on vascular smooth muscle cell ( VSMC ) and its mechanism. Methods The cells were cultured in DMEM supple-mented with 10% fetal bovine serum. MTT assay was used to examine cell proliferation. FCM was used to observe cell cycle. The expressions of PCNA, Cy-clinD1 and p-ERK1/2 were analyzed using Western blot. Results CurTn could inhibit the proliferation of VSMC and showed a certain amount-time relationship. What’ s more, CurTn could increase the G1 phase pro-portion of cell, decrease the S phase proportion and the expression level of PCNA protein. It was also found that CurTn significantly inhibit the protein expression of p-ERK1/2 and Cyclin D1 . Conclusion CurTn may inhibit the proliferation of VSMC via downregulating the expression of CyclinD1 and p-ERK1/2 .

OBJECTIVETo assess therapeutic effect of combined treatment of Chinese medicine and western medicine on optic atrophy complicated by cerebral palsy.

METHODSOne hundred and seventeen cases were divided into an observation group (n = 79) and a control group (n = 38). The control group were treated with routine western medicine treatment including neurotrophic drugs and high pressure oxygen, etc. and the observation group with acupuncture at Ganshu (BL 18), Pishu (BL 20), Chengqi (ST 1), etc. and injection of 0.2-0.3 mL Compound Danshen Injectio into Qiuhou (EX-HN 7), on the basis of the same treatment of western medicine as that in the control group. Fundus examination and the tracing body angle detection were conducted before and after treatment and the therapeutic effects were assessed in the two groups.

RESULTSThe total effective rate was 91.1% in the observation group and 60.5% in the control group with a significant difference between the two groups (P < 0.001); after treatment the angle of tracing body significantly increased in the two groups (P < 0.01) with the observation group better than the control group (P < 0.01).

CONCLUSIONThe combined treatment of Chinese medicine and western medicine is an effective therapy for optic atrophy complicated by cerebral palsy.

Acupuncture Therapy ; Cerebral Palsy ; complications ; Combined Modality Therapy ; Drug Therapy ; Female ; Humans ; Infant ; Male ; Optic Atrophy ; therapy

OBJECTIVETo investigate the effect of ginsenoside Rg1 on the microenvironment dependent differentiation of human mesenchymal stem cells (hMSCs) to vaso-endothelioid cells (VECs) in vitro.

METHODSThe in vitro differentiation of hMSCs to VECs were established adopting the in vivo environment simulated semi-permeable membrane separated non-contact co-culturing method. The mRNA expressions of endothelial markers, such as platelet endothelial adhesive factor-1 (CD31), vascular hemophillia factor (vWF) and vascular endothelial cadherin (VE-cadherin) were analyzed by RT-PCR; the protein expressions of CD31 and vascular endothelial adhesive factor-1 (VCAM1) were detected by fluorescence immunohistochemistry; structural identification for the endothelial characteristics of differentiated hMSCs were made under electron microscopy; and the percentage of CD31 expression in differentiated hMSCs was determined by flow cytometry to explore the effect of ginsenoside Rg1 on the differentiation.

RESULTSThe bone marrow mesenchymal stem cells co-cultured with mature endothelial membrane showed a microenvironment dependent capacity for differentiating to endothelium, with the morphological changes revealed starting from the 2nd week, showing cell body contraction, polygonal-shaped change; and at the 3rd week, the markedly speedily cell proliferation with elliptic or slabstone-like change of cells. High levels of classic endothelial cell markers, such as mRNA expressions of CD31, vWF, VE-cadherin, and protein expressions of CD31 and VCAM1, were shown; the typical weibel-palade body of endothelial cell was found in the differentiated cells. Moreover, percentage of CD31 expression in the differentiated hMSCs was increased after Rg1 treatment dose-dependently.

CONCLUSIONUnder the microenvironment of co-culture, hMSCs could differentiate into cells presenting the characteristics of endothelial cell in aspects of the morphology and ultrastructure of cells, as well as the gene and protein expressions of cell markers; ginsenoside Rg1 can promote the microenvironment dependent differentiation of hMSCs to VECs system in vitro.

Bone Marrow Cells ; cytology ; Cadherins ; metabolism ; Cell Differentiation ; drug effects ; Cells, Cultured ; Cellular Microenvironment ; drug effects ; Coculture Techniques ; Endothelium, Vascular ; cytology ; Ginsenosides ; pharmacology ; Humans ; Mesenchymal Stromal Cells ; cytology ; Panax ; chemistry ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; Vascular Cell Adhesion Molecule-1 ; metabolism ; von Willebrand Factor ; metabolism

Objective To establish a method for studying molecular mechanism of Rhubarb inhibiting anti-Yersinia pesti based on DNA microarray.Methods A whole genome DN A microarray containing 4005 annotated genes of Yersiniapesti Was used.The minimal inhibitory concentration(MIC)of Rhubarb to Yersiniapestiwas determined by liquid dilution method.The gene expression profile of Yersinia pesti was performed after the exposure to Rhubarb at a concentration of 10×MIC for 30 minutes.The total RNA extracted and purified from Yersinia pesti Was reversely transfected to cDNA and labeled by Cy3-Cy5 dye.The labeled probes were hybridized to the microarray anti the results were obtained by a laser scanner and the microarray data was confirmed by real-time quantitative RT-PCR.Results The platform of the DNA microarray-based bacteria transcriptional profile was established.A total of 498 genes of Yersinia pesti changed significantly in response to Rhubarb.Among them.358 genes were up-regulated,140 down-reguated.Conclusions The whole genome DNA microarray can be used in the studying of molecular anti-Yersinia pesti mechanism of Rhubarb.

Objective To investigate the possible role of interstitial cell of Cajal (ICE)in the pathogenesis of diabetic gastroparesis and the protective effect of"Xiaopi prescription (消痞方)".Methods Fifty healthy male Spregue-Dawley (SD) rats were randomly divided into 5 groups (each n=10):normal, model,and 3 Xiaopi prescription groups:low,middle and high dosages.Diabetes was induced by intravenous injection of alloxan,and equal amount of normal saline was intravenously injected in the normal group.Gastric lavage method was used to administer the traditional Chinese medicine decoction of Xiaopi prescription in corresponding amount (5,10 and 20 g?kg~(-1)?d~(-1)) in respective low,middle and high dosage groups.In the normal control group and diabetic model group,only equal amount of normal saline was administered into the stomach.Gastric emptying rate was measured by method of nutritious semisolid paste;c-kit positive cells of ICC were quantitatively measured with immunohistochemistry assay and computer image analysis system;c-kit mRNA positive cells were quantitatively measured with in situ hybridization and computer image analysis system.Results①Gastric emptying rate:The rate was significantly lower in the model group than that in the normal control group (P0.05),but higher than those in the model group and the low dosage group (all P0.05).②c-kit immmunohistochemistry:c-kit positive cell presented yellow in color,and its membrane was stained yellow,this kind of cells primarily were distributed around the neural plexus in the inter-space between the circular and longitudinal muscular fibers, and around the ganglionic cells forming"sheath-like"structure.The results of numbers of c-kit positive cells in the various groups:the number of the cells in the model group was significantly lower than that in the normal group (P0.05),but the numbers in the former two dosage groups were obviously higher than those in the model group (all P0.05),being significantly lower than that in the normal group,middle and high dosage groups (all P0.05),but obviously higher than those in the model group (all P0.05),being significantly lower than that in the normal,middle and high dosage groups (all P

Objective To investigate the risk factors of perivascular space change after brain tumor surgery.Methods According to the occurrence of postoperative perivascular space change,80 cases of glioma patients were divided into perivascular space and reconstruction group(observation group,n=38)and normal postoperative perivascular space group(control group,n=42).Compared the general data,sur-gery,tumor related indicators and postoperative complications of the two groups,and analyzed the influencing factors of the perivascular space changes after brain tumor surgery.Results In the observation group,the operation time of the patients was(95.38 ±9.21)min,which was significantly longer than(75.36 ±9.05)min in the control group.The intraoperative blood loss was(290.32 ±45.47)mL in the observation group,which was significantly more than(247.19 ±36.75)mL in the control group,and the difference was statistically significant (P<0.05).The tumor site located in the left hemisphere,tumor volume more than 40.0 cm3,high grade glioma,and proportion of patients with postoperative complications in the observation group were all higher than those of the control group,and the difference between the two groups was statistically significant(P<0.05).Multivariate Logistic regression analysis showed that age,tumor location,tumor volume,patho-logical grade and complications were significantly correlated with the changes of perivascular space after surgery(P<0.05).Conclusion Advanced age,tumor located in the left side of the brain,large tumor volume,severe pathology,postoperative epilepsy,chronic intracranial hy-pertension and other complications were the risk factors affecting the changes of perivascular space in patients with glioma.

OBJECTIVETo identify the epidemical characteristics of suicidal tendency among middle-school students in cities of China and to explore the main factors leading to suicidal tendency in adolescents.

METHODSMulti-stage cluster sampling method was used to select 9015 students in grades 1, 2, 3 and 4 respectively from 25 general middle schools in Beijing, Hangzhou, Wuhan and Urumqi of China in June 2006 and field investigation was carried out through "China Global School-based Student Health Survey (GSHS) Questionnaire".

RESULTSAmong the students in the four cities, the incidence rates of suicidal ideation were from 14.4% to 20.8% with an average of 17.4%. The incidence rates of suicidal plan were from 6.8% to 9.7% with an average of 8.2% and were different among cities. 15.0% of the boys had suicidal ideation and 6.7% of them made a suicidal plan comparing to 19.7% of girls having had suicidal ideation and 9.5% of them made a suicidal plan. The two kinds of suicidal tendency in girls were all higher than those in boys. City, age, gender, grade, days and type of being bullied, depression, close friends and having received health education on coping with stresses were factors influencing suicidal tendency of students. Days of being bullied and suicidal tendency showed a dose-response relation.

CONCLUSIONSuicidal tendency seemed common in middle-school students. Training on 'coping the issue' should be strengthened and harmonious environment should be improved in middle-schools.

Adolescent ; China ; epidemiology ; Data Collection ; Female ; Humans ; Incidence ; Male ; Students ; psychology ; Suicide ; psychology ; statistics & numerical data ; Urban Population

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Chromosome Aberrations ; Chromosomes, Human, Pair 8 ; Female ; Fibromatosis, Aggressive ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Middle Aged