Objective Detection of S-100B protein in different degree meconium pollution level in serum of children with value.MethodsIn 2012 June to 2013 December in our department were simple meconium stained amniotic fluid in term newborns in 73 cases, and set up for the observation group, and according to the amniotic fluid pollution degree is divided into 47 cases of amniotic fluid of Ⅰ-Ⅱdegree pollution group and 26 cases in grade Ⅲmeconium group during the same period, selected 20 cases without amniotic fluid contamination in term healthy newborns for the control group, the groups were compared in S-100B protein content difference.ResultsⅢmeconium stained amniotic fluid were within 6h serum S-100B was significantly higher than that in control group(P<0.01), and the degree of contamination of amniotic fluidⅠ-Ⅱgroup compared with the control group, no significant difference(P>0.05).Ⅲ meconium stained amniotic fluid in children with 72h also increased.ConclusionThird degree meconium stained amniotic fluid but normal Apgar score of newborns may still exist in clinical brain injury, so pay close attention to.

Melittin exhibits high antibacterial potency against drug-resistant bacteria. However, the clinical utility of melittin is limited by its serious hemolytic activity. Thus, the need for developing novel melittin analogues with high antimicrobial activity and low hemolytic activity has grown. We designed, synthesized, and evaluated 20 novel melittin analogues with varying hydrophobic, polar or positively charged amino acids. The results showed that 8 compounds had antimicrobial activity (MIC: 1-4 μg·mL^-1) against gram-positive pathogens equal to or better than that of melittin, and 16 compounds had low hemolytic activity (HC₅₀ ≥ 11.9 μg·mL^-1). Compounds 13 (MIC: 2-4 μg·mL^-1) and 15 (MIC: 1-2 μg·mL^-1) showed equal or better antimicrobial activity against both susceptible and resistant strains of Staphylococcus aureus and Enterococcus faecium compared to melittin (MIC: 4 μg·mL^-1). Compound 13 (HC₅₀: 24.0 ± 4.3 μg·mL^-1) displayed noticeably decreased hemolytic activity compared to melittin (HC₅₀: 5.3 ± 0.4 μg·mL^-1). This work established a base for further study on the structure-activity relationships and structure-toxicity relationships of melittin.

OBJECTIVETo explore the effect of ligustrazine on the migration of bone marrow mesenchymal stem cells (BMSCs) and protein expressions of matrix metalloproteinase-2 and-9 (MMP-2 and MMP-9) in vitro.

METHODSBMSCs were in vitro isolated and cultured using whole bone marrow adherent method, and phenotypes [surface positive antigens (CD29 and CD90) and negative antigens (CD34 and CD45)] identified using flow cytometry. BMSCs were divided into the blank control group, 25, 50, 100 µmol/L ligustrazine group, and the GM6001 group (100 µmol/L ligustrazine +MMPs inhibitor GM6001 ). The migration of BMSCs was tested by Transwell chamber test and wound healing assay after treated with ligustrazine for 24 h. The protein expressions of MMP-2 and MMP-9 were detected by Western blot.

RESULTSThe third passage BMSCs grew well in uniform morphology. The expression rate of CD29, CD90, CD34, and CD45 was 96.9%, 97.3%, 0.2%, and 3.0%, respectively. Compared with the blank control group, the number of migrated cells and relative distance of cell invasion increased, and the protein expressions of MMP-2 and MMP-9 were elevated in each ligustrazine group (P < 0.05, P < 0.01). Compared with 100 µmol/L ligustrazine group, the number of migrated cells and relative distance of cell invasion decreased in 25 and 50 µmol/L ligustrazine groups and the GM6001 group (P < 0.01). Protein expression of MMP-2 decreased in 25 and 50 µmol/L ligustrazine groups (P < 0.01).

CONCLUSIONLigustrazine could promote the migration of BMSCs in vitro, and its mechanism might be related to up-regulating expression levels of MMP-2 and MMP-9 protein.

Cell Movement ; Cells, Cultured ; Hematopoietic Stem Cells ; cytology ; drug effects ; Humans ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Pyrazines ; pharmacology ; Up-Regulation

OBJECTIVETo improve tooth pulp electrical stimulation rat model to study analgesic effect of drugs.

METHODSExpose lower mandible and incisor and isolate them with rubber dam, two holes with distance 1.5 mm were drilled below the cemento-enamel junction by a thin diamond bur in 22 adult male rats anesthetized with sodium pentobarbital. A pair of insulated stainless stimulating electrodes were inserted through the holes and knotted to fix them. Drilled holes were filled with zinc polycarboxylate cement. Finally, the stimulating electrodes were directed to the top of the skull. Pain threshold of rats was determined by an electrical stimulator under awake, free conditions to evaluate the stability of the model and the analgesic effect of drugs.

RESULTSA valid tooth pulp pain model were set up successfully in 22 rats. Rats regain consciousness at 1 to 2 hours after operation. The inserted electrodes keep efficiency for 6 weeks. Neither tissue morphology change nor inflammation cell infiltration can be found in tooth pulp after 6 weeks under the light microscope. Threshold voltages remained constant under the repeat electric stimulations within 300 min (P > 0.05, CV < 15%) until 4 weeks (P > 0.05, CV < 15%). The dose-dependent and time-dependent analgesic effects of capsaicin were showed in this model.

CONCLUSIONThe improved tooth pulp electrical stimulation rat model is easily performed and shows constant electrical stimulation-induced pain threshold and is suitable for research on analgesic effect of drugs.

Animals ; Dental Pulp ; Electric Stimulation ; Incisor ; Male ; Pain ; Rats

In order to find the cardiotonic constituents of lateral roots of Aconitum carmichaelii Debx., the investigation was carried out. Silica gel column chromatography, Sephadex LH-20, medium-pressure MCI and reverse phase ODS column chromatography were used to separate the 90% EtOH extract of the lateral roots of Aconitum carmichaelii Debx. The structures of the isolated compounds have been identified by chemical properties and spectroscopic analyses. Ten compounds were isolated and their structures were elucidated as benzoic acid-5-hydroxy-2-benzoyl-amino methyl ester (1), honokiol (2), pinoresinol (3), salicylic acid (4), p-hydroxy-cinnamic acid (5), songorine (6), karakoline (7), mesaconitine (8), hypaconitine (9) and 14-benzoylhypaconitine (10), separetely. Compound 1 is a new compound and its structure has been established by NMR, HR-ESI-MS, UV, IR and X-Ray. Compound 2-5 are isolated from the lateral roots of Aconitum carmichaelii Debx. for the first time.
Aconitum ; chemistry ; Cardiotonic Agents ; chemistry ; isolation & purification ; Plant Roots ; chemistry

Background The effect of chronic stress on cognitive functions has been one of the hot topic in neuroscience. But there has been much controversy over its mechanism. Such single stressor applied in the past could not simulate complicated living circumstances that people confronted with. The aim of this study was to investigate the effects of chronic multiple-stress on learning and memory as well as on the levels of calcium/calmodulin-dependent protein kinase II (CaMKII), calmodulin (CaM) mRNA, and cAMP-response element binding protein (CREB) mRNA in the hippocampus of rats. Methods The rats were divided randomly into stressed and control groups. The stressed group was given chronic multiple-stress for 6 weeks to set up a chronic multiple-stressed model. The rats' performance of spatial learning and memory was tested using Morris Water Maze (MWM) and Y-maze. Meanwhile, the expressions of CaMKII, CaM mRNA and CREB mRNA of rats' hippocampus were detected by immunohistochemistry, Western blot and reverse transcription-polymerase chain reaction (RT-PCR), respectively. In addition, the width of synaptic cleft and the thickness of post-synaptic densities (PSD) were observed in the hippocampal CA3 region of rats by electron microscopy. Results After exposure to chronic multiple-stress for 6 weeks, the ability of learning and memory of the stressed group was higher than that of the control group (P<0.05, P<0.01). The width of synaptic cleft was smaller and the thickness of PSD was larger in the hippocampal CA3 region of the stressed group than in that of the control group (P<0.01). The CaMKII immunostaining of the stressed group was stronger than that of the control group in the stratum radiatum and oriens of the hippocampal CA1 and CA3, especially in the stratum oriens. Quantitative analysis indicated that the expression of CaMKII, CaM mRNA, and CREB mRNA in the hippocampus of the stressed group was higher than that of the control group (P<0.05, P<0.01). Conclusions The capacity of learning and memory can be enhanced after chronic multiple-stress. The increased levels of CaMKII, CaM mRNA, and CREB mRNA may contribute to the enhancing effect of chronic multiple-stress on learning and memory.

The objective was to identify some biochemical and physical properties for fusion protein IL6D24-PE40KDEL. Edman degradation, SDS-PAGE, peptide mass fingerprinting, Western blot and MTT were used for identification of the protein. The results showed that the sequence of N-terminus is Met-Ile-Asp-Lys-Gln-Ile, Met was added because of prokaryotic expression system; Western blot revealed that the purified protein could react with IL6 and PEA antibody. The purified protein IL6D24-PE40KDEL could kill the multiple myeloma cell lines U266 expressing high affinity IL6R, but it could not kill the cell lines CEM which not expressed IL6R; The molecular weight was 58.7 kD measuring by SDS-PAGE; peptide mass fingerprinting (PMF) confirmed that the construction of IL6D24-PE40KDEL was correct. A novel protein by Peptident database in EXPASY web site was identified. In conclusion, IL6D24-PE40KDEL is a new targeting protein with bioactivity of specific killing effect.
ADP Ribose Transferases ; chemistry ; metabolism ; pharmacology ; Amino Acid Sequence ; Blotting, Western ; Cell Line, Tumor ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Exotoxins ; chemistry ; metabolism ; pharmacology ; Humans ; Interleukin-6 ; chemistry ; metabolism ; pharmacology ; Molecular Sequence Data ; Pseudomonas aeruginosa ; genetics ; metabolism ; Recombinant Fusion Proteins ; chemistry ; metabolism ; pharmacology ; Sequence Analysis, Protein ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVETo study the inhibitory effects of celecoxib, a cyclooxygenase-2 inhibitor, on the proliferation of hepatocellular carcinoma cells.

METHODSThe in vitro inhibitory effects of celecoxib at different concentrations and for different treatment time lengths on human liver cancer cell line SMMC-7,721 were observed with MTT assay, and flow cytometry was performed to detect the cell cycle changes. The in vivo tumor inhibition effect of celecoxib was evaluated in Kunming mice bearing transplanted tumor derived from liver cancer cell line H22 transplantation.

RESULTCelecoxib significantly inhibited the in vitro growth of human liver cancer cell line SMMC-7721 in a time- and dose-dependent manner. A 36-hour celecoxib treatment (40 micromol/L) resulted in decreased SMMC-7721 cell proliferation and an increase of the cell percentage in G1 phase from 44.7% to 49.9% with decreased cell percentage in S and G(2)/M phases from 55.4% to 50.1%. In the mice bearing H22 transplanted tumor, celecoxib showed significant inhibitory effect on the growth and local metastasis of the transplanted tumor.

CONCLUSIONCelecoxib can inhibit the proliferation of different liver cancer cell lines both in vitro and in vivo, and therefore may serve as an important candidate drug for prevention and treatment of hepatocellular carcinoma.

Animals ; Carcinoma, Hepatocellular ; drug therapy ; physiopathology ; Celecoxib ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclooxygenase 2 Inhibitors ; pharmacology ; therapeutic use ; Female ; Humans ; Liver Neoplasms ; drug therapy ; physiopathology ; Mice ; Neoplasm Transplantation ; Pyrazoles ; pharmacology ; therapeutic use ; Sulfonamides ; pharmacology ; therapeutic use

OBJECTIVETo explore the relationship between CD8+ T-cell CD28 molecular expression in peripheral blood and TCM type in patients with chronic aplastic anemia (CAA).

METHODSUsing flow cytometry to detect the CD28 expression in 45 in-patients or out-patients and 24 healthy subjects for control. And the relation with TCM type was analyzed from the immunological aspect.

RESULTS(1) The levels of CD8, CD28, CD8+ CD28+ expression and CD8+ CD28+/CD8+ CD28- were all higher in the CAA patients than those in the healthy subjects (P < 0.05 or P < 0.01). (2) The levels of CD28, CD8+ CD28+ expression and CD8+ CD28+/CD8+ CD28- were all higher in the CAA patients of Shen-Yin deficiency type than those in the CAA patients of Shen-Yang deficiency type (P < 0.05 or P < 0.01).

CONCLUSION(1) The abnormal high expression of peripheral blood co-stimulatory molecules CD28 suggested CD28 disorder may play an important role in immuno-pathogenesis of CAA. (2) The levels of peripheral CD28, CD8+ CD28+ expression and CD8+ CD28+/CD8+ CD28- can be taken as an objective indexes for TCM typing of CAA, which was disordered more severe in patients of Shen-Yin deficiency type than in those of Shen-Yang deficiency type.

Adult ; Anemia, Aplastic ; classification ; diagnosis ; immunology ; CD28 Antigens ; biosynthesis ; CD8-Positive T-Lymphocytes ; immunology ; Chronic Disease ; Diagnosis, Differential ; Humans ; Kidney Diseases ; immunology ; Male ; Medicine, Chinese Traditional ; Yang Deficiency ; immunology ; Yin Deficiency ; immunology

OBJECTIVETo determine whether long-term treatment of attention deficit hyperactivity disorder (ADHD) with methylphenidate influences the growth in height and weight of children.

METHODSAnalyses were performed on 146 school age children (126 boys) diagnosed as ADHD and treated with methylphenidate [0.27-0.64 mg/(kg.day)] for methylphenidate group and 29 children with ADHD who did not receive any medication for ADHD (controls). These children were followed-up for 2-4 years. Changes in height and weight after long-term treatment with methylphenidate were recorded and the factors affecting growth of height, weight, and height velocity were analyzed.

RESULTSThe change of difference between patients' height and mean height in methylphenidate group and controls was (-1.86 +/- 0.82) cm (paired t test, t = 27.335, P < 0.001) and (-0.26 +/- 0.51) cm (P < 0.05), respectively; the change of height standard deviation score (SDS) in methylphenidate group and controls was -0.14 +/- 0.23 SD (paired t test, t = 7.326, P < 0.001) and +0.05 +/- 0.10 SD (P < 0.05), respectively. When the height change and height SDS change in methylphenidate group and controls were compared by using independent-samples T-test, the t value was -10.078 and -4.262 respectively, P for both was < 0.001. Both of bivariate correlation analysis and stepwise multiple-regression analysis indicated that the duration of treatment contributed significantly to the variance in change of height (P < 0.001); but age, sex, DSM-IV type, NJ22 degree and dose of methylphenidate did not contribute significantly to the variance of height. The mean height velocity from 1st to 4th year was 4.28 cm/year, 4.90 cm/year, 4.98 cm/year and 4.95 cm/year, respectively. With Friedman test, Chi-square = 253.673, P < 0.001. The change of difference of patients' weight to weight for height after methylphenidate was (-0.14 +/- 1.25) kg (paired t test, t = 1.326, P > 0.05).

CONCLUSIONSmall but significant deceleration of height velocity is the identified long-term side effect of methylphenidate, the magnitude of height deficit is related to duration of treatment. The height velocity was significantly attenuated in the first year. Methylphenidate had no significant influence on weight.

Attention Deficit Disorder with Hyperactivity ; drug therapy ; Body Height ; drug effects ; Body Weight ; drug effects ; Case-Control Studies ; Central Nervous System Stimulants ; adverse effects ; therapeutic use ; Child ; Child Development ; Female ; Humans ; Male ; Methylphenidate ; adverse effects ; therapeutic use ; Regression Analysis