Aim To study the effect of gamma-hydroxybutyric acid receptor(GHBR ) on neuronal apoptosis suffering from focal cerebral ischemia-reperfusion injury in rats. Methods The male Sprague-Dawley rats weighing 240～280 g were randomly divided into seven groups: sham operation group(sham), ischemia-reperfusion group(Isc/R) ,NCS-356 160、320、640 ?g?kg-1 group(N1、N2、N3),NCS-382 640+NCS-356 640 ?g?kg-1 group(NCS-382+N3),and nimodipine 600 ?g?kg-1 group(Nim).The middle cerebral artery occlusion(MCAO) model invented by Zea Longa with modifications was adopted. The experiment was divided into two parts after ischemia reperfusion for 24h:In the first part,we measured the cerebral expression of Bax, Bcl-2, Caspase-3 by immuneohistochemical method. In the second part,we measured neuronal apoptotic rate by flow cytometry in the ischemic cortex region. Results The expression rate of Bcl-2 and Bcl-2/Bax ratio of N1,N2,N3 and Nim groups were all higher than that of Isc/R group(P

Animals ; Cadmium Chloride ; toxicity ; Dose-Response Relationship, Drug ; Estradiol ; toxicity ; Female ; Organ Size ; drug effects ; Rats ; Rats, Sprague-Dawley ; Uterus ; drug effects ; pathology

Aim To observe the effects of the L-dopa methyl ester (LDME) on the pattern visual evoked potentials (P-VEP) and the expression of c-fos mRNA in neurons of the visual cortex of kittens with strabismic amblyopia, and explore the therapeutic effect of L-dopa methyl ester on amblyopia and its action mechanism.Methods 30 normal kittens were randomly divided into 6 groups: low dose of L-dopa methyl ester (20 mg·kg~(-1)), medium dose of L-dopa methyl ester (40 mg·kg~(-1)), and high dose of L-dopa methyl ester (80 mg·kg~(-1)),positive control (L-dopa 40 mg·kg~(-1)),normal control, and model control group.The surgery for producing iatrogenic convergent strabismus was performed on 4 weeks old kittens(normal control group excluded). After the confirmation of the development of amblyopia by pattern visual evoked potential,L-dopa methyl ester,L-dopa and normal saline were given to the corresponding animals, respectively. The P-VEP of amblyopia eyes was observed after one month, and the technique of in situ hybridization was used to detect the expression of c-fos mRNA.Results L-dopa methyl ester could reduce obviously the length of P100 peak latency of the cat strabismic amblyopes, and increase the amplitude of P100. The positive staining cells of strabismic cat visual cortex were less than those of normal cats, whose difference was significant (P<0.01).Positive staining cells in the treatment group were significantly increased when compared with that of the model group (P<0.01).Conclusion L-dopa methyl ester can significantly improve the conduction and sensory function in the model of strabismic amblyopia cats. The mechanism may be related to the increased amount of L-dopa methyl ester into the cerebral cortex and regulation of the expression of c-fos mRNA.

Objective To evaluate PDCA circle in training of cooperation quality in endoscopic nurse.Methods PDCA circle was applied to train endoscopic nurses.Knowledge of endoscopic nursing and techniques as well as operators' satisfaction with endoscopic examination were compared before and after PDCA circle training.Results Nursing scores increased from 92.40 to 96.00 with statistical significance (P ＜ 0.05).Performance scores increased from 93.03 to 98.13 (P ＜ 0.05).Operators' satisfaction increased from 81.3％ to 96.9％.Conclusion PDCA circle could improve not only endoscopic nursing but also the quality of endoscopic training.

OBJECTIVEPrevious studies have shown that hydrogen sulfide (H2S) plays key roles in a number of biological processes, including vasorelaxation, inflammation, apoptosis, ischemia/reperfusion and oxidative stress, which are involved in the pathogenesis of myocarditis. This study aimed to examine the expression of cystathionine-γ-lyase(CSE)/H2S pathway in mice with viral myocarditis.

METHODSSix-week-old inbred male mice were randomly assigned to control (n=25) and myocarditis group (n=30). The myocarditis and the control groups were inoculated intraperitoneally with 0.1 mL 10-5.69TCID50/mL CVB3 or vehicle (PBS) alone respectively. Ten mice were sacrificed 4 and 10 days after injection. Blood and heart specimens were harvested for measuring the content of serum H2S and the H2S production rates in cardiac tissues. Heart sections were stained with hematoxylin and eosin. Immunohistochemisty was used to detect the CSE protein expression in the heart.

RESULTSIn the myocarditis group, the serum H2S content and H2S production rates in cardiac tissues were significantly higher than those in the control group 4 and 10 days after injection (P<0.05). The expression of CSE protein in the heart in the myocarditis group was also significantly higher than that in the control group (P<0.05).

CONCLUSIONSCSE and its downstream production H2S increase in mice with acute viral myocarditis. The increased expression of CSE/H2S pathway might be involved in the pathogenesis of viral myocarditis.

Animals ; Coxsackievirus Infections ; etiology ; Cystathionine gamma-Lyase ; analysis ; Enterovirus B, Human ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Hydrogen Sulfide ; metabolism ; Killer Cells, Natural ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Myocarditis ; etiology

Objective:To observe the effect of acupuncture in regulating ubiquitin-proteasome pathway (UPP),and discuss the action of acupuncture in intervening heroin-induced brain damage.Methods:Thirty male Sprague-Dawley (SD) rats were divided into a control group,a model group and an acupuncture group by using the random number table.Rats in the model and acupuncture groups received intramuscular heroin injection for successive 8 d at a progressively increased dose.Afterwards,the injection was suspended for 5 d for withdrawal.The heroin relapse rat model was established by repeating the drug addiction and withdrawal process for 3 times.The control group followed the step of the model establishment,but was given intramuscular injection of normal saline at the stage of addiction and no intervention at the stage of withdrawal;the model group was given intramuscular heroin injection at a progressively increased dose at the addiction stage and no intervention at the withdrawal stage;the acupuncture group was dealt in the same way as the model group at the addiction stage,but received acupuncture at Baihui (GV 20) and Dazhui (GV 14) at the withdrawal stage,with the needles retained for 30 min each time,1 session a day,for successive 5 d.On the 39th day,brain tissues were extracted from the hippocampus and ventral tegmental area (VTA) of the three groups of rats.The apoptosis of brain nerve cells was detected by using terminal deoxynucleotidyl transferase-mediated nick and labeling (TUNEL).The mRNA and protein expressions of ubiquitin (Ub),ubiquitin protein ligase (E3) and 26S were examined by immunohistochemistry and quantitative real-time polymerase chain reaction (RT-qPCR).Results:Compared with the model group,rat's hippocampus and VTA in the acupuncture group showed significantly fewer cells positively stained by TUNEL staining (P＜0.01),and its mRNA and protein expressions of Ub,E3,26S were significantly lower (P＜0.01).Conclusion:Reducing nerve cell apoptosis and regulating the mRNA and protein expressions of Ub,E3 and 26S in rat's hippocampus and VTA are possibly one of the action mechanisms of acupuncture in intervening heroin-induced brain damage.

OBJECTIVETo investigate the down-regulated TRAF6 gene expression and its effects on proliferation and apoptosis in multiple myeloma (MM) cells.

METHODSDetection of TRAF6 expression were conducted by RT-PCR and Western blot in MM cell lines of KM3, U266, RPMI8226 and primary cells from patients. RPMI8226 cell lines were transfected with siRNA of TRAF6. The efficiency of transfection was identified by using of fluorescence microscope, RT-PCR, and Western blot. The levels of proliferation were analyzed by CCK-8 method under the different concentrations of siRNA. Apoptosis rate were detected with Hoechst33258/PI double staining by flow cytometry. Apoptosis related proteins Bcl-2, BAX, and NF-κB signal pathway were observed before and after siRNA transfection by Western blot.

RESULTSThe levels of TRAF6 mRNA and protein in MM cell lines, especially in primary myeloma cells, were significantly higher than those in controls. After transfected with 50 nmol/L siRNA in RPMI8226 cells, the relative level of TRAF6 mRNA (0.49±0.24) was significantly lower than that in non-transfected group (1.87±0.23) and idling group (1.74±0.35). The proliferation rate of siRNA transfected cells decreased with dose dependence (P<0.01). The apoptosis rates increased from 11.20% (before transfection) to 51.82% (after transfection), accompanied by down-regulated Bcl-2 protein, NF-κB signal pathway (p-p65 and p52), and up-regulated BAX protein.

CONCLUSIONTRAF6 expression was high in myeloma cells. TRAF6 siRNA could inhibit proliferation of myeloma cells and induce apoptosis mediated by NF-κB classical and alternative pathway in myeloma cells.

Case-Control Studies ; Cell Proliferation ; Down-Regulation ; Female ; Gene Expression ; Humans ; Male ; Multiple Myeloma ; metabolism ; pathology ; TNF Receptor-Associated Factor 6 ; genetics ; metabolism ; Tumor Cells, Cultured

OBJECTIVETo introduce a new way of treatment for elbow stiffness and instability.

METHODS30 cases of elbow trauma were treated with mobile hinged Orthofix elbow fixator, 26 of them were followed up. There are 15 male patients and 11 female patients. The average age is 32.6 years old. The 26 injured elbows involved 12 cases on left side and 14 cases on right side; Sixteen cases on the dominant side and 10 cases on the non-dominant side. Sixteen cases had elbow stiffness, 5 cases had acute radial head fracture and posterior dislocation of the elbow, 3 of cases had Monteggia's fracture dislocation, and 2 cases had neglected posterior dislocation of the elbow.

RESULTSThe average follow up time was 6 months (3 - 12 month). The mean time of fixation with fixator was 8.5 weeks (6 - 11 weeks). The mean ROM of the 16 cases of elbow stiffness was (37.5 +/- 0.8) degrees before operation, and (96.5 +/- 0.6) degrees operation, with a significant difference (P < 0.05). The mean Mayo elbow score was (69.5 +/- 1.7) before operation, compared with (82.8 +/- 1.6) after operation, with a significant difference (< 0.05). For the other 10 cases (5 cases with acute radial head fracture and posterior dislocation of the elbow, 3 cases with Monteggia's fracture dislocation, 2 cases with neglected posterior dislocation of the elbow), the average ROM of the elbow flexion-extension was 95 degrees (65 degrees - 150 degrees ); The average range of flexion was 117 degrees; the average loss of extension was 22 degrees; the average pronation was 76 degrees (20 degrees - 90 degrees ), the average supination was 75 degrees (15 degrees - 90 degrees ). Nine of the cases achieved anatomic reduction and proved by X-ray. The mean Mayo elbow score was 84 (49 - 96). Three cases were rated excellent, 4 good, 2 fair and 1 poor. The rate of excellent and good was 70% (7/10). Nine cases had no pain or mild pain, and did not need analgesic. Eight cases returned to their former work, 5 of the cases had complications.

CONCLUSIONSThe mobile hinged elbow external fixator have following advantages: (1) distraction of the articular space and enhance fracture healing; (2) allow early movement of the elbow during healing; (3) provide stable environment for the healing after arthrolysis and reconstruction. Mobile hinged elbow external fixator can achieve successful result in the treatment of elbow stiffness or unstable fracture dislocation of elbow.

Adolescent ; Adult ; Aged ; Elbow Joint ; injuries ; surgery ; External Fixators ; Female ; Follow-Up Studies ; Fracture Fixation ; methods ; Humans ; Joint Dislocations ; complications ; surgery ; Joint Instability ; surgery ; Male ; Middle Aged ; Monteggia's Fracture ; complications ; surgery ; Radius Fractures ; complications ; surgery ; Treatment Outcome

OBJECTIVETo explore the effect and mechanism of Poly I:C in inducing growth inhibition and apoptosis of human hepatocellular carcinoma SMMC-7721 cells.

METHODSSMMC-7721 cells were treated with different doses of Poly I:C for 24, 48, and 72 h, and the cell growth inhibition rate was analyzed with CCK-8 assay. The cell cycle and the apoptosis were analyzed using flow cytometry with Annexin-V and PI staining, and quantitative RT-PCR analysis were used to detect the expression of TLR3, TRIF, and IFN-beta mRNA in cells.

RESULTSIn the cells exposed to Poly I:C at low, moderate, and high doses, the inhibitory rates was the highest in high-dose Poly I:C group, and at a given Poly I:C dose, prolonged exposure resulted in significantly increased cell growth inhibition rate (P<0.05). Flow cytometry showed that Poly I:C induced cell apoptosis in a time- and dose-dependent manner and significantly increased the percentage of G1-phase cells as compared with that in the control group. The mRNA level of TLR3, TRIF, and IFN-beta were also increased following Poly I:C treatment in comparison with the control group.

CONCLUSIONPoly I:C can induce significant growth inhibition and apoptosis of SMMC-7721 cells in a dose- and time-dependent manner possibly by causing cell cycle arrest and TLR3 signaling pathway activation that leads to IFN-beta production and cell apoptosis.

Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Humans ; Interferon-beta ; genetics ; metabolism ; Liver Neoplasms ; pathology ; Poly I-C ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Receptors, Cholecystokinin ; metabolism ; Signal Transduction ; Toll-Like Receptor 3 ; genetics ; metabolism

AIMTo investigate the level of immune response and the immune mechanism of the single-dose hepatitis B surface antigen (HBsAg)-poly (d, l)-lactide-co-glicolide acid (PLGA) microspheres in BALB/c mice.

METHODSThree kind of HBsAg-PLGA microspheres, HBsAg-PLGA50/50-COOH microspheres, HBsAg-PLGA75/25 microspheres and HBsAg-PLGA50/50 microspheres, were prepared by double emulsion microencapsulation technique used three kinds of PLGA with different L/G ratio. The single-dose of HBsAg-PLGA microspheres was subcutaneously injected into BALB/c mice at the dose of 7.5 microg HBsAg per mouse. The conventional aluminum-adjuvant vaccine was subcutaneously injected at 0, 1 and 2 month as positive control. In certain time interval, the induced immune level of total antibody was detected by enzyme linked immunosorbent assay (ELISA). For subclass of IgG antibody and cytokines studies, the dose of HBsAg was 2.5 microg per mouse.

RESULTSThe HBsAg-PLGA microspheres could successfully induce a humoral immune response in BALB/c mice. Compared with the conventional aluminum-adjuvant vaccine, the antibody response of the HBsAg-PLGA50/50-COOH microspheres was significantly lower than the group received three injections of aluminum-adjuvant vaccine (P < 0.01) except for a higher priming response during the early 6 weeks. The results were ascribed to the relatively rapid degradation charactics of PLGA50/50-COOH polymer. The immune response for the HBsAg-PLGA50/50 microspheres and HBsAg-PLGA75/25 microspheres were comparable to the group administered with aluminum-adjuvant vaccine (P > 0.05) which was due to the sustained degradation of PLGA50/50 and PLGA75/25 polymer.

CONCLUSIONThe HBsAg-PLGA microsphere is a promising candidate for the controlled delivery of a vaccine which does not require multiple injections.

Animals ; Delayed-Action Preparations ; Dose-Response Relationship, Immunologic ; Drug Carriers ; Female ; Hepatitis B Antibodies ; blood ; Hepatitis B Surface Antigens ; administration & dosage ; chemistry ; immunology ; Hepatitis B Vaccines ; administration & dosage ; immunology ; Immunization ; Immunoglobulin G ; blood ; Interferon-gamma ; metabolism ; Interleukin-2 ; metabolism ; Interleukin-5 ; metabolism ; Lactic Acid ; Mice ; Mice, Inbred BALB C ; Microspheres ; Polyglycolic Acid ; Polymers ; Random Allocation ; Rats ; Rats, Wistar