To investigate the therapeutic effect of high-dosage gamma-aminobutyric acid (GABA) on acute tetramine (TET) poisoning, 50 Kunming mice were divided into 5 groups at random and the antidotal effects of GABA or sodium dimercaptopropane sulfonate (Na-DMPS) on poisoned mice in different groups were observed in order to compare the therapeutic effects of high-dosage GABA with those of Na-DMPS. Slices of brain tissue of the poisoned mice were made to examine pathological changes of cells. The survival analysis was employed. Our results showed that both high-dosage GABA and Na-DMPS could obviously prolong the survival time, delay onset of convulsion and muscular twitch, and ameliorate the symptoms after acute tetramine poisoning in the mice. Better effects could be achieved with earlier use of high dosage GABA or Na-DMPS. There was no significant difference in prolonging the survival time between high-dose GABA and Na-DMPS used immediately after poisioning. It is concluded that high-dosage GABA can effectively antagonize acute toxicity of teramine in mice. And it is suggested that high-dosage GABA may be used as an excellent antidote for acute TET poisoning in clinical practice. The indications and correct dosage for clinical use awaits to be further studied.
Acute Disease ; Antidotes/*administration & dosage ; Antidotes/therapeutic use ; Bridged Compounds/*poisoning ; Random Allocation ; Rodenticides/*poisoning ; Unithiol/therapeutic use ; gamma-Aminobutyric Acid/*administration & dosage ; gamma-Aminobutyric Acid/therapeutic use

AIM　To study the effects of Phe-Met-Arg-Phe-NH2 (FMRFa) on Na+/Ca2+ exchange and its specificity for Na+/Ca2+ exchange in rat ventricular myocytes. METHODS　Na+/Ca2+ exchange current and other currents of ion channels were measured using whole cell voltage clamp techniques. RESULTS　A dose-related inhibition of tetrapeptide FMRFa on Na+/Ca2+ exchange was observed in rat ventricular myocytes. Inward and outward INa+/Ca2+ were inhibited by 60.1% and 56.5%, respectively, at highest concentration (100 μmol*L-1) and its IC50 were 20 μmol*L-1 and 34 μmol*L-1 in inward and outward INa+/Ca2+, respectively. Inward and outward INa+/Ca2+ were inhibited 38.7% and 34.9%, respectively, at FMRFa 5 μmol*L-1. FMRFa 5 μmol*L-1 and 20 μmol*L-1 did not affect L-type calcium current, sodium current, transient outward current and inward rectifier potassium current. CONCLUSION　These data indicate that FMRFa is a specific inhibitor of Na+/Ca2+ exchange in intact rat ventricular myocytes.

Objective To study the inhibition of tumor necrosis factor-alpha(TNF-α)cytokine expression of BV-2 cells induced by hypoxia-reoxygenation injury by siRNA targeting TLR4 gene via the RNAi mechanisms.Method BV-2 mouse microglial cell line was cultured in six-well plates and randomly divided into group N(nor-mal group),group H(hypoxia-reoxygenation),group T(hypoxia-reoxygenation+TLR4-siRNA transfected group),group C(hypoxia-reoxygenation+pEGFP-H1/control-siRNA transfected group)and group B(hypox-ia-reoxygenation+pEGFP-H1 transfected group).Group H,group T,group C and group B were cultured in hy-poxia condition for 3 h followed by reoxygenation for 24 h.The plasma was transfected into BV-2 cells mediated by lipofectamine 2000.The efficiency of transfection were detected by flow cytometry to observe the expression of EGFP.RT-PCR method was used to detect the level of mRNA of TLR4 or NF-кB p65.Westem blot methed was used to test the expression of TLR4 protein.and ELISA was used to test the level of TNF-α in the supernatants.Analysis of variances was used for statistical analysis.Results The expression of EGFP gene waa;(67.58±7.16)% after transfection by flow cytometry analysis.Compared to group N,the TLR4 mRNA,NF-кB p65 mR-NA,TLR4 protein level and the TNF-α quantity in group H,group T,group C and group B increased after the hy-poxia-reoxygenation treatment(P＜0.01).While the expression of the TLR4 mRNA,NF-кB p65 mRNA,TLR4 protein level and the TNF-α quantity in the group T down-regulated compared to group H,group C and group B(P＜0.01).And there were no changes in group C,group B and group H about observation index(P＞0.05).Conclusions The siRNA targeting TLR4 mRNA could inhibit the inflammatory reaction released by BV-2 cells in-duced by hypoxia-reoxygenation stimulation.

Objective To observe the changes in concentrations of pulmonary surfactant SP-A/B in lung tissue during acute lung injury (ALI) /acute respiratory distress syndrome (ARDS) induced by acute paraquat poisoning (PQP) after the treatment with metabolic antioxidant,lipoic acid,and to explore the potential involvement of TNF-α in ALI/ARDS as well as to discuss the assumed protective mechanisms of lipoic acid against acute lung injury.Methods Sixty-six male Sprage-Dawley rats were randomly (random number) divided into 3 groups,namely control group (NS,n =6),paraquat poisoning group (PQ,n =30),paraquat + lipoic acid treatment group (LA,n =30).Then both group PQ and group LA were further divided separately into five subgroups,namely 3,6,12,24 and 48 h subgroups (n =6 in each subgroup).After rats sacrificed,the lung tissues were selected,and after HE staining,histological changes were observed under light microscope.Histopathological changes were inflammation and fibrosis in models successfully established.The lung tissues were also taken for tests of SOD and MDA levels.Specimens of whole blood 0.8 mL without anticoagulant were taken from tail vein of rats for determining the TNF-α level.The expressions of SP-A mRNA and SP-B mRNA were measured with RT-PCR from total RNA of the lung tissue.Results ① HE staining showed that the histopathological changes were milder in LA group than that in PQ group.② There were significant differences in MDA and SOD levels between different intervals both in intergroups and intragroup except the groups of 3 hours (P ＜ 0.01).③ Likewise,the significant differences in the levels of TNF-α were also present between three groups and between different intervals (P＜0.01).④ The significant differences in SP-A mRNA and SP-B mRNA amplification ratio existed between three groups at the same interval (P ＜ 0.01),but those differences between different intervals in group PQ were of statistical significance (P ＜ 0.05).And those differences between diffirent intervals in group LA were statistically significant (P ＜0.01).Conclusions Lipoic acid in acute paraquat poisoning could lessen lung tissue damage,which might be directly dominated by the levels of tumor necrosis factor,and in turn indirectly affect the content of pulmonary surfactant,thereby reducing pulmonary edema and improving lung compliance,then protecting the lung tissues.

ObjectiveTo study the quality of life of patients with hepatolenticular degeneration (HLD)and analyze the influencing factors.Methods287 patients with HLD and 51 health people were investigated by World Health Organization quality of life assessment instrument brief version (WHOQOL-BREF),Symptom Checklist 90 (SCL-90),Life Satisfaction Index A (LSIA) and variance analysis,t-test and multiple linear regression analysis were analyzed the influencing factors.Results①Scores of WHOQOL-BREF:physical domain(54.64 ± 17.11 ),psychological domain ( 52.09 ± 15.83 ) in patients with HLD were lower than those in the health people (67.30 ± 12.66,58.90 ± 12.75 ) with statistically significant difference (P ＜ 0.01 ) ; social domain ( 51.35± 17.18),the domain of environment(53.54 ± 16.67) in patients with HLD were lower than those in the health people (57.53 ± 14.99,58.42 ± 10.55 ) with statistically significant difference (P ＜ 0.05 ).②The quality of life of the patients with HLD was influenced by LSIA,total score of SCL-90,the attitude toward the doctors,economic status,the attitude toward the disease,residence with statistically significant difference (P ＜ 0.0l ).ConclusionThe quality of life in patients with HLD is lower than that in health people and much factors influence it,so it is necessary to take multi-facet interventions to improve their quality of life.

Objective To study the expression of S100A8 and the relationship between S100A8 and Toll-like receptor 4 (TLR4) in focal cerebral ischemia reperfusion (I/R) injury.Methods C3H/HeJ mice with TLR4 gene mutation (n =30) and C3H/HeN with normal TLR4 gene mice (n =30) were divided into 4 groups at random (random number),namely C3H/HeJ model group (n =18),C3H/HeJ control group (n =12),and C3H/HeN model group (n =18).C3H/HeN control group (n =12).Middle cerebral artery was occluded to make I/R model in mice by using thread embolism method.Brain tissues were collected after ischemia for one hour and reperfusion for 12 hours.Stroke outcome was evaluated by determination of infarct volume of brain tissue and assessment of neurological scores.And brain injury after cerebral I/R was observed by optical microscope after TTC and HE staining.The immunofluorescence technique and RT-PCR were used to determine the protein level and expression of S100A8 mRNA in damaged brain tissues.Results Compared with C3H/HeN model mice,TLR4-deficient model mice (C3H/ He J) had lower infarct volumes and better outcomes of neurological function after resuscitation for 12 hours.Compared with control groups,the expression of S100A8 mRNA and level of S100A8 protein increased greatly in damaged brain tissues of model mice after I/R injury.In addition,model mice with lacked TLR4 (C3H/HeJ) had lower expression of I/R-induced S100A8 mRNA than C3H/HeN mice in model group,indicating that the close relationship between the levels of S100A8 and TLR4.Conclusions S100A8 interaction with TLR4 might be involved in brain damage and in inflammation triggered by I/R injury.

AIMTo explore the changes of mRNA and protein expressions of heart-type fatty acid binding protein (H-FABP) in rat ischemic myocardium at different intervals ischemia.

METHODS60 SD male rats weighing 250-350 g, were randomly divided into one sham-operated group and five study groups (group A1, A2, A3, A4, A5, the left coronary artery of rats has been ligated for 1 h, 2 h, 4 h, 6 h, 12 h respectively). Myocardil samples from infarct zone, ischemic and non-ischemic zone, were obtained for histology examination, and the mRNA for H-FABP in ischemic myocardial tissue were determined by RT-PCR. Serum free fatty acid(FFA) was determined by colorimetric method.

RESULTSCompared to sham hearts, H-FABP mRNA expression were significantly decreased in ischemia zone of AMI rat hearts (P < 0.05), especially in rats underwent 4 h ischemia and 6 h ischemia (P < 0.01). Serum FFA were significantly increased in AMI rats relative to sham rats (P < 0.05).

CONCLUSIONSignificant down-regulated heart-type fatty acid binding protein after myocardial ischemia might play an important role in myocardial injury and energy metabolism disorder.

Animals ; Down-Regulation ; Fatty Acid-Binding Proteins ; genetics ; metabolism ; Male ; Myocardial Infarction ; metabolism ; Myocytes, Cardiac ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Objective:To study the anti-depression mechanism of Chaiyuwendan Decoction(CYWDD).Methods:Rat models were established by separation and chronic unpredictable mild stress(CUMS).Ethology of rats were detected by open-field test and sucrose consumption test,contents of monoamine neurotransmitters were detected by HPLC-ECD.Results:Compared with normal group,the weight,the frequency of crossing,rearing and the contents of consumption to sucrosum water in depressive disorder rats decreased significantly(P

Objective To establish a fast and effective gene diagnosis method for Wilson's disease(WD) patients by double PCR-SSCP technology.Methods We amplificated exon8 and exon12 of ATP7B gene by double PCR from genomic DNA of 140 unrelated WD patients and 30 normal controls,then used SSCP technology to screen them.At last we identificated these patients' mutation features by direct sequencing.Results No abnormality shift was found in 30 controls.In 140 patients,we found 7 types of abnormal mobility shifts in 66 cases(47.14%).In subsequent direct sequencing,mutation rate of Arg778Leu/Gln was 37.14%(52/140),and Thr935Met was 12.86%(18/140).Conclusion Double PCR-SSCP diagnosis technology is a effective method which can improve diagnosis rate for Wilson disease.

OBJECTIVEThis study aims to examine the expression of nuclear factor κB (NF-κB)/B cell lymphoma-2 (Bcl-2) signal pathway and glycogen synthase kinase 3β (GSK-3β) in oral squamous cell carcinoma (OSCC) and provide references for the early diagnosis and prognosis evaluation of OSCC.

METHODSA total of 55 cases of OSCC and 10 cases of paracan- cerous mucosa were examined in this study. Their expressions of GSK-3β, NF-κB and Bcl-2 were detected using the SP me- thod immunohistochemistry. The correlation between their expression in OSCC and the clinical and pathological peculiarity of OSCC was analyzed.

RESULTSThe positive expression of GSK-3β, NF-κB, and Bcl-2 in OSCC were significantly higher than that in paracancerous mucosa (P < 0.01). The expression of GSK-3β, NF-κB, and Bcl-2 had no obvious relationship with patient's age, sex, and clinical stages of the disease (P > 0.05). The expression of Bcl-2 was significantly correlated with the degree of tumor differentiation (P < 0.05), whereas the expression of GSK-3β and NF-κB in OSCC had no obvious relation- ship with the degree of tumor differentiation (P > 0.05). Strong positive correlations were observed among the expressions of GSK-3β, NF-κB, and Bcl-2 (P < 0.05).

CONCLUSIONThe positive expression of GSK-3β, NF-κB, and Bcl-2 in OSCC are sig- nificantly higher than that in paracancerous mucosa. Detecting GSK-3β, NF-κB, and Bcl-2 in OSCC may have implications in the early diagnosis and prognosis evaluation of OSCC.

Apoptosis ; Cell Differentiation ; Glycogen Synthase ; Glycogen Synthase Kinase 3 ; Humans ; Lymphoma, B-Cell ; Mouth Neoplasms ; NF-kappa B ; Neoplasms, Squamous Cell ; Signal Transduction