Background: Synaptic plasticity contributes to nociceptive signal transmission and modulation, with calcium/ calmodulin-dependent protein kinase II (CaMK II) playing a fundamental role in neural plasticity. This research was conducted to investigate the role of CaMK II in the transmission and regulation of nociceptive information within the nucleus accumbens (NAc) of naïve and morphine-tolerant rats. Methods: Randall Selitto and hot-plate tests were utilized to measure the hindpaw withdrawal latencies (HWLs) in response to noxious mechanical and thermal stimuli. To induce chronic morphine tolerance, rats received intraperitoneal morphine injection twice per day for seven days. CaMK II expression and activity were assessed using western blotting. Results: Intra-NAc microinjection of autocamtide-2-related inhibitory peptide (AIP) induced an increase in HWLs in naïve rats in response to noxious thermal and mechanical stimuli. Moreover, the expression of the phosphorylated CaMK II (p-CaMK II) was significantly decreased as determined by western blotting. Chronic intraperitoneal injection of morphine resulted in significant morphine tolerance in rats on Day 7, and an increase of p-CaMK II expression in NAc in morphine-tolerant rats was observed. Furthermore, intra-NAc administration of AIP elicited significant antinociceptive responses in morphine-tolerant rats. In addition, compared with naïve rats, AIP induced stronger thermal antinociceptive effects of the same dose in rats exhibiting morphine tolerance. Conclusions This study shows that CaMK II in the NAc is involved in the transmission and regulation of nociception in naïve and morphine-tolerant rats.

OBJECTIVETo investigate the effect of the selective PI3K inhibitor and MEK inhibitor on KRAS and PTEN co-mutated non-small cell lung cancer cell line NCI-H157 and the relevant mechanisms.

METHODSNCI-H157 was cultured routinely and treated with different concentrations of the two inhibitors. Cell proliferation was detected by MTT cell cycle assay. Based on the MTT results the cells were divided into four groups: the control group, PI3K inhibitor group (GDC-0941, 0.5 and 5.0 µmol/L), combination group I (0.5 µmol/L AZD6244 + 0.5 µmol/L GDC-0941) and combination group II (5.0 µmol/L AZD6244 + 5.0 µmol/L GDC-0941). Colony formation assay was performed to detect colony formation efficiency. The cell cycle and apoptosis were analyzed by flow cytometry. The expression of protein related to apoptosis was tested with Western blot.

RESULTSCell growth was inhibited by the two inhibitors. Combination groups led to stronger cell proliferation inhibition: combination group Ishowed synergistic effect of their actions and combination group II showed an additive effect; in both groups, there were decreased colony number [(77.2 ± 1.54)/well vs (61.50 ± 2.12)/well, P < 0.01] and [(51.00 ± 4.00)/ well vs (22.50 ± 3.53)/well, P < 0.01]; and enhanced apoptotic ratios [(18.30 ± 0.82)% vs (21.32 ± 0.56)%, P < 0.01] and [(27.14 ± 1.58)% vs (42.45 ± 4.42)%, P < 0.01]. In addition, compared to the PI3K inhibitor alone group, the NCI-H157 cells in the combination groups showed increased G0/G1 phase and decreased S phase (P < 0.01). Western blotting showed that the combination groups demonstrated significantly decreased expression of cyclin D1 and cyclin B1, increased p21 and cleaved PARP and decreased bcl-2/bax ratio, compared to the PI3K inhibitor only group.

CONCLUSIONThe combined inhibition of PI3K (AZD6244) and MEK (GDC-0941) has synergistic effects on the proliferation of NCI-H157 cells, but such effects appear to be in a dose-dependent manner.

Apoptosis ; drug effects ; Benzimidazoles ; administration & dosage ; pharmacology ; Carcinoma, Non-Small-Cell Lung ; genetics ; pathology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin B1 ; metabolism ; Cyclin D1 ; metabolism ; Dose-Response Relationship, Drug ; Drug Synergism ; Humans ; Indazoles ; administration & dosage ; pharmacology ; Lung Neoplasms ; genetics ; pathology ; Mitogen-Activated Protein Kinase Kinases ; antagonists & inhibitors ; metabolism ; Mutation ; PTEN Phosphohydrolase ; genetics ; Phosphatidylinositol 3-Kinases ; antagonists & inhibitors ; metabolism ; Poly(ADP-ribose) Polymerases ; metabolism ; Proto-Oncogene Proteins ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Proto-Oncogene Proteins p21(ras) ; metabolism ; Signal Transduction ; Sulfonamides ; administration & dosage ; pharmacology ; bcl-2-Associated X Protein ; metabolism ; ras Proteins ; genetics

OBJECTIVETo evaluate the amount of blood loss and the efficacy of clotting factor in controlling blood loss during total knee arthroplasty.

METHODSThe medical documents of 18 patients with haemophilic arthritis (HA) secondary to haemophilia A and 19 patients with osteoarthritis (OA) were retrospectively reviewed. Demographic data,functional and hematological test results,the amount of blood loss and transfusion,and complications were analyzed.

RESULTSThe median amounts of total and external blood loss were 2240 ml(1892-3415 ml) and 1326 ml(934-2256 ml)in the HA group, which were significant higher than those in the OA group [1746 ml(1259-2246 ml)and 846 ml (504-1217 ml), respectively]. The median amounts of external blood loss in the two groups were 680 ml(370-1330 ml)and 730 ml(200-1190 ml)and there was no significant difference(p=0.620). Moreover, more patients in the HA group required blood transfusion (84.2% vs. 47.4%), and more red cells were transfused per patient in the HA group (2.3 U vs. 0 U).

CONCLUSIONSThe total blood loss and hidden blood loss are higher in the HA patients than in OA patients during total knee arthroplasty, although the external blood loss is basically the same. Management with more clotting factor may decrease the blood loss in HA patients.

Adolescent ; Adult ; Arthritis ; etiology ; surgery ; Arthroplasty, Replacement, Knee ; Hemophilia A ; complications ; Humans ; Male ; Middle Aged ; Osteoarthritis, Knee ; surgery ; Postoperative Hemorrhage ; Retrospective Studies ; Young Adult

OBJECTIVETo determine the physicochemical properties of the mutanase of Trichoderma harzianum isolated from China and to study the influence of mutanase on the adherence of oral Streptococci and the structure of oral biofilms.

METHODSSix fungal strains belonging to Trichoderma were tested for mutanase production in the same cultural condition, the strain producing the highest mutanase activity was studied further and the pH and temperature optimum of the enzyme was determined. The RT-PCR method was used to obtain the gene coding for mutanase and the product was cloned to pMD18-T simple vector for sequencing. Inhibition effects of mutanase on the adherence of Streptococcus sobrinus OMZ176, Streptococcus sobrinus 6715, Streptococcus mutans MT8148 were studied by adherence test. The optical sectioning of biofilms with or without mutanase supplementation were analyzed by confocal laser scanning microscopy (CLSM).

RESULTSThe highest enzymatic activity was achieved by Trichoderma harzianum Th1, the maximum activity was at pH 5.5 and at 40 degrees C. The nucleotide sequence was 92% homology with that of a known gene coding a mutanase (GenBank accession No. AJ243799). The adherence of Streptococcus sobrinus OMZ176, Streptococcus sobrinus 6715, Streptococcus mutans MT8148 was significantly inhibited by mutanase. Compared with control, the biofilms with mutanase supplementation had lower height and sparser structure.

CONCLUSIONSThe mutanase from Trichoderma harzianum Th1 can inhibit the adherence of oral Streptococci and had an influence on the structure of oral biofilms.

Bacterial Adhesion ; drug effects ; Biofilms ; Glycoside Hydrolases ; chemistry ; physiology ; Streptococcus mutans ; drug effects ; Streptococcus sobrinus ; drug effects ; Trichoderma ; enzymology ; pathogenicity

AIM: To establish an animal model of experimental autoimmune myocarditis (EAM) in BALB/c mice and to investigate the expression and significance of Toll-like receptor 3 in mouse EAM. METHODS: BALB/c mice were immunized with cardiac myosin extracted from porcine ventricular myocardium covered by complete freund's adjuvant (CFA) on 0 d and 7 d, then divided into immunized with CFA only. Serum and myocardium samples were collected at 14 d and 21 d after the first immunization. HE staining was used to identify the areas of inflammation. The myosin IgG antibody was examined by indirect ELISA assay. The changes of TLR3 protein and mRNA expression in myocardial tissue were measured by immunohistochemistry and real time-PCR. RESULTS: Compared to control group, immunohistochemistry results showed that there was positive expression of TLR3 in the myocardium of mice with EAM and the mRNA of TLR3 were more than 20 times (P<0.05). The expression of interferon beta mRNA in EAM group was more than 14 times as many as basal expression, that of tumor necrosis factor alpha was more than 18 times (P<0.05). CONCLUSION: The expression of Toll-like receptor 3 in myocardium is up-regulated in experimental autoimmune myocarditis. The inflammatory response to cardiac myosin may associate with the TLR3 signal transduction pathway.

Objective To investigate the abdomen CT characteristics of patients with ischemic bowel disease (ICBD).Methods CT imaging data of ICBD patients from January 2008 to December 2013 in the Chinese PLA General Hospital were retrospectively analyzed to find CT imaging features in intestinal lesions associated with ICBD death.Results In CT imaging analysis,151 patients including acute superior mesenteric artery thromboembolism (ASMATE,n=51),acute superior mesenteric vein thrombosis (ASMVT,n=53),non-occlusive mesenteric ischemia (NOMI,n=8),chronic mesenteric ischemia (CMI,n=10) and ischemic colitis (IC,n=29) were divided into survival group (n=115) and death group (n=36).In comparison with the survival group,the death group had higher incidence of abdomenal effusion,portomesenteric gas,pneumatosis intestinalis and pneumoperitoneum (P＜0.001,P＜0.001,P＜0.001,P=0.003).Conclusion The ascites,portomesenteric vein gas,pneumatosis intestinalis and pneumoperitoneum in bowel CT may be associated with the death of ICBD patients.

Objective To investigate the effects of ABCB1 genetic polymorphism on the dose(D) adjusted blood concentration(C/D) of tacrolimus (TAC) and renal function in renal transplant recipients with cytochrome P450 3A5 * 3/* 3 (CYP3A5 * 3/* 3) genotype during the stable period.Methods Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was employed to detect CYP3A5 * 3 and ABCB1 genotypes;allele,genotype frequencies and Hardy-weinberg equilibrium were analysed by SHEsis software.The effects of ABCB1 polymorphism on the C/D of TAC and serum creatinine (Scr) were analyzed using SPSS 18.0 software.Results For renal transplant recipients with CYP3A5 * 3/* 3 genotype,the C/D of tacrolimus in CC genotype of ABCB1 C1236T were significantly lower than that of CT + TT genotypes (P ＜ 0.05).The Scr of GG + GA + AA + GT + AT genotypes of ABCB1 G (A) 2677T were significantly lower than that of TT genotype (P ＜ 0.05).The Scr of CC + CT genotypes of ABCB1C3435T were significantly lower than that of TT genotype (P ＜ 0.05).Conclusion ABCB1 C1236T polymorphism has an effect on the C/D of tacrolimus in renal transplant recipients with CYP3A5 * 3/* 3 genotype during the stable period.Both ABCB1 G (A) 2677T and C3435T polymorphism influence the renal function of those transplant recipients.