OBJECTIVETo observe analgesic effect of electroacupuncture ( EA) on rats with chronic inflammatory pain and its regulatory mechanism on ispilateral dorsal root ganglion (DRG) and spinal dorsal horn (SDH) Mas-related G protein-coupled C receptor (MrgprC).

METHODSTotally 40 healthy male SD rats were divided into 4 groups according to random number table, i.e., the normal (N) group, the model (M) group, the acupuncture (Acu) group, the EA group, 10 rats in each group. The model of chronic inflammatory pain was established by subcutaneous injecting 0. 1 mL complete Freund's adjuvant (CFA) into right hind paw. Paw withdrawal thresholds (PWTs) were measured before modeling, at day 1, 3, 5, 7, and after CFA injection, respectively. Expression levels of MrgprC in ispilateral DRG and SDH were detected by Western blot. The content of bovine adrenal medulla 22 (BAM22) in SDH was detected by immunohistochemical assay.

RESULTSCompared with N group at each time point, PWTs significantly decreased in M group (P <0. 01). Compared with M group, PWTs significantly increased at day 5 of EA and after EA in EA group (P < 0.05, P < 0.01). Compared with Acu group at each time point, post-EA PWTs significantly increased in the EA group (P < 0.05). Compared with N group, expression of MrgprC in ispilateral DRG and ratio of BAM22 positive cells in ispilateral SDH increased in M group (P < 0.01). Compared with M group, expression of MrgprC in ispilateral DRG and ratio of BAM22 positive cells in ispilateral SDH increased in the EA group (P < 0.05).

CONCLUSIONEA had favorable analgesic effect on chronic inflammatory pain induced by CFA, and its mechanism might be possibly associated with up-regulating MrgprC expression in ispilateral DRG and BAM22 content in ispilateral SDH.

Analgesia ; Animals ; Electroacupuncture ; Enkephalins ; metabolism ; Freund's Adjuvant ; Ganglia, Spinal ; drug effects ; Inflammation ; chemically induced ; drug therapy ; Male ; Pain Management ; methods ; Peptide Fragments ; metabolism ; Posterior Horn Cells ; drug effects ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Objective To explore the effect of gaseous signal molecule hydrogen sulfide(H2S)on the expression of lung Muc5ac and Th1/Th2 cytokines in ovalbumin(OVA)-induced asthma rats and explore the effect of H2S on asthma.Methods Twenty-six healthy SD rats were randomly divided into 3 groups:control group(Group A,n=8),asthma group(Group B,OVA induced,n=9)and NaHS group(Group C,OVA induced rats treated with NaHS,n=9).Twenty-four hours after treatment,rats were anatomized to measure serum interleukin(IL)-4,interferon-? and the levels of infiltration into inflammatory cells around bronchus were observed,which were scored with the optical microscope.The expression of lung Muc5ac was determined by immunohistochemical staining.Results The score of the levels of infiltration into inflammatory cells around bronchus expressed by median was 1 score in group A,4 score in group B and 2 score in group C.There were dramatically statistics significance among the 3 groups(H=13.75 P

Objective To explore the clinical correlation of the variation of plasma homocysteine (HCY), melatonin (MLT) and ulceative colitis (UC). Methods The clinical data of 112 UC patients was collected, and 110 normal healthy persons as control. The level of plasma HCY and MLT was detected by high pressure liquid chromatography-fluorescence detection (HPLC-FD) method. The level of plasma folate ( FA) and vitamin B12 was detected by enzyme-linked immunosorbent assay (ELISA) method. The correlation of these four indexes and UC was analyzed. Results The serum level of HCY in UC patients was significantly higher than that in normal healthy persons [(11. 27± 7.26) μmol/L vs (8. 19±4. 81) μmol/L, P = 0. 000]. The serum level of MLT in UC patients was significantly lower than that in normal healthy persons [(49. 06 + 31. 40) pg/ml vs (64. 28±41. 16) pg/ml,P=0. 008]. The serum level of FA in UC patients was significantly lower than that in normal healthy persons [(7. 64 + 1.95) nmol/L vs (9. 14 + 1.23) nmol/L, P = 0. 005]. The serum level of vitamin B12 in UC patients was significantly lower than that in normal healthy persons [(108. 64 ±32. 22) pmol/L vs (112. 64±33. 33) pmol/L, P = 0. 004]. There was no correlation between plasma HCY, MLT and UC disease activity degree, range, disease duration, erythrocyte sedimentation rate (ESR), or C reactive protein (CRP) in UC patients. There was no significant correlation between MLT and HCY in UC patients. Conclusions The serum level of HCY is higher in UC patients than that in normal control, and MLT is lower than that in normal control. However there is no significant correlation between them.

Objective To investigate the inhibitory effect and mechanism of rosiglitazone on chemokines secretion in renal tubular epithelial cells (HK-2) stimulated by lipopolysaccharide (LPS). Methods Cells were divided into four groups: control (CON), LPS (1 mg/L),rosiglitazone (10 μmol/L), rosiglitazone (10 μmol/L) +LPS (1 mg/L). MCP-1 and IL-8 expression was measured using real time PCR and ELISA. PPARγ was knockdown by RNAi to investigate whether the inhibitory effect of rosiglitazone was PPARγ-dependent or -independent. The NF-κB in nucleus was detected by Western blotting. The DNA binding activity of NF-κB was determined by electrophoretic mobility shift assay. Results Compared with CON group, the expressions of IL-8 and MCP-1 were increased by (4.30±0.45) and (4.80±1.29) times in mRNA level, (1.39±0.18)and (2.11 ±0.47) times in protein level, respectively, in LPS-stimulated HK-2 cells (P＜0.05).Application of rosiglitazone followed by LPS significantly reduced IL-8 and MCP-1 secretion compared with LPS group (decreasing by 66.37% and 71.88% in mRNA levels, while 41.68% and 47.87% in protein levels) (P＜0.05). In pcDNATM 6.2-GW/EmGFP-miPPARγ transfected cells, IL-8and MCP-1 only were decreased by 18.16% and 16.83% in mRNA level, while 11.39% and 11.86%% in protein level in rosiglitazone pretreated group, showing no significant difference compared with LPS group. Rosiglitazone did not block NF-κB nuclear translocation while significantly inhibiting the DNA binding activity of NF-κB. Conclusions Rosiglitazone inhibits the expressions of MCP-1 and IL-8 via a PPARγ-dependent mechanism in HK-2 cells, resulting from inhibition the DNA binding activity of NF-κB.

Objective To study the role of vagusism on diethytlnitrosamine (DEN)-induced hepatocellular carcinoma in rats and to explore its mechanism.Methods The rat vagus nerve was labeled with 1,1'-dioctadecyl-3,3,33 -tetramethylindocarbocyanine perchlorate (DiI) tracer.The rats were divided into three groups:cancer induced rat models (cancer-induced group,n =20),cancer-induced and vagal detachment models (combined group,n =20) and normal rats (negative group,n =20),HE staining was performed on the liver tissue biopsied from the three groups of rats.The pathological grading score was evaluated and statistically analyzed.Immunohistochemistry (IHC) staing was used to determine the expression of vagal neurotransmitters and related receptors in liver tissue.Results The optimal concentration (1 mg/ml) and fixation duration with formalin (4 days) of DiI tracer marker vagal nerve demonstrated superior distribution and density of vagus nerve in the liver tissue.The incidence of hepatocellular carcinoma was significantly higher in the cancer-induced group (75％) than combined group (15％),the difference was statistically significant (P ＜ 0.05).IHC staining results of acetllcholine showed a significant difference between cancerinduced group (9.10) and combined group (2.30) or negative group (2.95),the difference was statistically significant (P ＜ 0.05).The M1 receptor expression was significantly higher in cancer-induced group (6.00) than combined group (2.30) and negative group (2.55),the differences were statistically significant (P ＜ 0.05).There was no significant difference of M2 receptor expression among the cancer-induced group (1.30),combined group (1.25) and negative group (1.35) (P ＞ 0.05).The M3 receptor expression detected with IHC staining shown there were statistical differences among cancer-induced group (8.95),combined group (6.30) and negative group (3.60) (all P ＜ 0.05).Conclusion The vagus nerve may play an important role in the development of DEN-induced hepatocellular carcinoma in rats.

Background Sweep pattern visual evoked potential (SVEP) is an objective method of visual test.There is a clear correlation between SVEP acuity and subjective vision,but they are not identical.Recent studies showed that new regression method can improve the accuracy of SVEP acuity. Objective This trial was to investigate and compare the outcome between amplitude-spatial frequency (A-SP) regression method and amplitudelogVA (A-logVA) regression method in extrapolating the SVEP acuity.Methods SVEP was recorded in 113 eyes of 64 subjects using GT-2000 ( Guo Te,China) with the gratings of 10 different spatial frequency from 0.99 to 12.89 cpd as stimulus.The 1 13 eyes included cataract,glaucoma,corneal disease,optical neuropathy,retinal disease,ocular trauma,refractive error and normal eyes.The correlation were analyzed of SVEP acuity,decimal visual acuity and LogMAR visual acuity.The response were averaged and DFT on the monitor display.SVEP acuity was calculated by extrapolating 0 response amplitude.Results The correlation indices of decimal visual acuity curves obtained by the A-logVA function was 0.663,and that obtained by the A-SP function was 0.705.The positive correlation was seen between subjective decimal visual acuity and A-logVA decimal visual acuity (r =0.540,P＜ 0.01 ) and between subjective decimal acuity and decimal acuity calculated by the A-SP regression method (r=0.620,P＜0.01 ).SVEP decimal acuity calculated by the A-SP function regression method was significantly different from the that calculated by the A-logVA function regression method (Z =-8.688,P＜0.01 ).And the correlation indices of LogMAR visual acuity curves obtained by the A-logVA function was 0.733 and that obtained by the A-SP function was 0.715.The positive correlation was found between the subjective LogMAR acuity and that calculated by the A-SP regression method (r=0.700,P＜ 0.01 ) and between the subjective LogMAR acuity and LogMAR acuity calculated by the A-logVA regression method (r=0.710,P＜0.01 ).SVEP LogMAR acuity from A-SP function regression method was significantly different from the LogMAR acuity from A-logVA function regression method (Z=-8.748,P＜0.01 ).No significant differences of VA LogMAR were found in gender,eyes,type of disease and age(x2 =2.171,P=0.338;x2 =0.976,P=0.614;x2 =6.032,P=0.420;x2 =14.720,P=0.257 ).Conclusions SVEP can obtain the visual outcome in human.The amplitude-logVA function regression method is more accurate in extrapolating SVEP acuity.

OBJECTIVETo evaluate the impact of a new CD44 variant on invasion of human breast cancer cell line MCF-7, and its possible mechanisms.

METHODSThe full length cDNA encoding CD44v17 was obtained from the total RNA isolated from the MCF-7/ADR cells by reverse transcript-polymerase chain reaction (RT-PCR) and subcloned into pMD19-T vector. The CD44v17 gene sequence and reading frame were confirmed by two restriction enzymes and nucleotide sequencing, and then inserted into the eukaryotic expression vector pcDNA3.1. The pcDNA3.1-CD44v17 was transfected into MCF-7 cells by Lipofectamine. The changes of MMP-2 and MMP-9 expression at gene and protein levels were detected by RT-PCR and gelatin zymography, respectively. The number of the cells through the artificial matrix membrane in every group was counted to compare the change of the invasive ability regulated by CD44 variant. The ERK and p-ERK were investigated by Western blotting to approach the molecular mechanisms of MMP-2 and MMP-9 expression regulated by CD44 variant.

RESULTSThe new gene sequence was successfully cloned into recombinant vector pcDNA3.1 and identified by the two restriction enzymes. It was confirmed that the reconstructed plasmid contained the sequence of CD44 gene variant which was composed of 1 to 4 exons, 16 to 17 exons, and 1 to 205 bases of 18 exons. The new gene sequence was sent to NCBI for publication and obtained the registered number FJ216964. The up-regulated levels of the CD44 gene mRNA and protein were respectively detected by RT-PCR and flow cytometry in MCF-7 cells transfected with pcDNA3.1-CD44v17. The invasiveness of the cells and the activity of MMP-2 and MMP-9 were clearly activated by hyaluronic acid (HA) treatment and blocked by CD44 neutralizing antibody. Pretreated MCF-7/CD44v17 cells with the neutralizing antibody against CD44 and the inhibitor of MAPKs signaling pathway strongly block the expression of p-ERK.

CONCLUSIONA new CD44 gene variant has been found in adriamycin-resistant human breast cancer MCF-7/ADR cells. The expression vector pcDNA3.1-CD44v17 has been cloned and constructed successfully. HA can be integrated with CD44 variant and then regulates the expression of MMP-2 and MMP-9, which increases the invasion ability of MCF-7 cells through the Ras/MAPK signaling pathway.

Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; Drug Resistance, Neoplasm ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Genetic Vectors ; Humans ; Hyaluronan Receptors ; genetics ; metabolism ; Hyaluronic Acid ; pharmacology ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Neoplasm Invasiveness ; Phosphorylation ; Plasmids ; Protein Isoforms ; RNA, Messenger ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Signal Transduction ; Transfection

0.05).Conclusion Febrile seizures seldomly cause severe neurological damage.

OBJECTIVETo evaluate the clinical efficacy and safety of aripiprazole in the treatment of childhood tic disorders (TD) by a meta analysis.

METHODSA systematic search for randomized controlled trials (RCTs) on the efficacy and safety of aripiprazole in the treatment of childhood TD that were published between January 2000 and August 2014 was conducted. A Meta analysis on the selected RCTs was conducted using Review Manager 5.2 software.

RESULTSSix RCTs involving 551 TD patients were enrolled. There were no significant differences in the efficacy between aripiprazole and traditional drugs for treatment of TD either by the end of follow-up visit or at 2 weeks, 4 weeks and 8 weeks after treatment. The subgroup analysis results indicated that aripiprazole had the same efficacy for the treatment of TD as traditional drug haloperidol. Aripiprazole had a lower incidence of extrapyramidal reactions than haloperidol (P<0.05), but the overall incidence of side effects of aripiprazole was not lower than traditional drugs for treatment of TD.

CONCLUSIONSThe available evidence suggests that aripiprazole has the same curative effect in the treatment of childhood TD compared with the traditional drugs. However, it is difficult to draw a firm conclusion that aripiprazole is a safer drug in the treatment of childhood TD.

Antipsychotic Agents ; therapeutic use ; Aripiprazole ; adverse effects ; therapeutic use ; Humans ; Tic Disorders ; drug therapy

OBJECTIVETo develop an HPLC method for the determination of eleutheroside B in As1 (the extract fraction from Acanthopanax senticosus, which has a good effect of antifatigue).

METHODThe antifatigue effect of Asl was evaluated by mice burden swimming testing. One compound was isolated by silica gel, Sephadex LH-20 column chromatography and preparative high performance liquid chromatography. The structure was identified by physicochemical properties and spectral evidences. Eleutheroside B in Asl was determined by HPLC. Chromatographic conditions included Diamonsil C18 column (4.6 mm x 250 mm, 5 microm) and the mobile phase consisting of a mixture of acetonitrile-water-ethanoic acid (10: 90: 0.01). The UV detection wavelength was set at 344 nm.

RESULTAs1 showed an excellent antifatigue activity; One compound was isolated from As1 and its structure was identified as Eleutheroside B; The calibration cure was linear in the range of 0.104-20.8 microg (r = 0.9999), the average recovery was 97.68%, RSD 1.4% (n=6).

CONCLUSIONThis HPLC method is simple,accurate and reliable.

Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Eleutherococcus ; chemistry ; Fatigue ; drug therapy ; physiopathology ; Glucosides ; analysis ; Humans ; Male ; Mice ; Phenylpropionates ; analysis ; Swimming