Objective To explore the change of hippocampal neurons cholinergic receptor in rats with vascular dementia(VD).Methods VD rat models were established by employing improved method of Pulsinelli's four-vessel occlusion.1 month later,the abilities of learning and memory of VD models were tested by Morris water maze.The activities of acetylcholinesterase (AChE) in plasma and hippocampus were detected by the improved Ellman's colorimetric methods.The expressions of hippocampal nicotinic acetylcholine receptor (nAChR) subunits ?3,?4 and ?7 proteins and mRNA were detected respectinely by Western Blotting and RT-PCR,respectively.The results were compared with sham operation group.Results Compared to the sham operation group,the escape latency of Morris water maze test and the first time crossing platform in VD group were significantly prolonged,the number of crossing platform was decreased (all P

Objective To investigate the effect of exogenous hydrogen sulfide (H 2 S) on H2 S concentration and cystathionine β-synthase (CBS) expression in hippocampus in a rat model of vascular dementia (VaD). Methods A rat model of VaD was induced by using the modified four -vessel occlusion. The rats were divided into sham operation, model, low -dose and high-dose NaHS groups using the random number table method. They were further redivided into one day, seven -day, and 30-day subgroups according to the time after modeling. After modeling respectively, NaHS 30 μmol/kg and 100 μmol/kg were injected intraperitoneally every day in the low -dose and high-dose NaHS groups. The normal saline was injected intraperitoneally every day in the sham operation group and the VaD model group. Morris water maze test was used to evaluate the learning and memory ability of the rats. The expression of CBS in hippocampus was detected by real-time fluorescent polymerase chain reaction. Western Blotting was used to detect expression of CBS protein in hippocampus. Results Morris water maze test showed that the escape latencies of the model group, low -dose and high-dose NaHS groups were prolonged significantly compared with the sham operation group (P <0.05); the times of crossing the platform were decreased significantly compared with the model group (P <0.05); and the escape latencies were shortened significantly in the low -dose and high-dose NaHS groups compared with the model group ( P <0.05). The H2 S content in hippocampus was decreased significantly in the model group, low -dose and high-dose NaHS groups compared with the sham operation group, but the low -dose and high-dose NaHS group was significantly higher than that in the model group (all P <0.05). The expression of CBS mRNA and protein in the model, low -dose and high-dose NaHS groups was significantly lower than that of the sham operation group (all P < 0.05), and there was no significant difference between the low -dose and high-dose NaHS groups and the model group. Conclusions Exogenous H2 S may improve the learning and memory ability of the VaD rats. It may be associated with the increased H2 S content in hippocampus. However, it has no effect on CBS expression.

The successful establishment of animal models of cardiac arrest-cardiopulmonary resuscitation (CA-CPR) undoubtedly provided an important basis for exploring the method of cardiopulmonary resuscitation (CPR) and advanced cardiovascular life support (ACLS). However, pathophysiology varied with the etiology of cardiac arrest (CA). Therefore, preparation of similar animal models according to etiology was the basis for pathophysiological changes research. Compared with other animals, the rabbits had both the advantages of large and small animals, so they became common research object for the CA-CPR model. This paper reviewed the common methods of animal models of CA-CPR in rabbits. In this review, the methods, criteria, advantages, disadvantages and precautions of each model were analyzed, which would provide useful reference for CPR researchers.

Objective To study the changes of PI3K/Akt/GSK3β signaling pathway during resuscitation with neck cooling in order to explore the relationship between the protective effect of neck cooling and the phosphorylation of PI3K/Akt and GSK3β.Methods Thirty rabbits were randomly(random number) divided into five groups, and models of cadiac arrest were induced by ventricular fibrillation(VF, the positive electrode in the right ventricle and negative pole on the apex of heart) for 4 min.In sham group,a electrode was placed into right ventricle without electric current conducted, and CA was not induced.The rabbits were sacrificed and specimens were taken at 24 hours after modeling.In normothermia treat group(NT group),resuscitation was carried out to restoration of spontaneous circulation(ROSC),and the rabbits were sacrificed and specimens were taken at 24 hours after modeling.In intra-arrest therapeutic hypothermia group (IATH group), rapid neck cooling was initiated at the same time with CPR,and the target brain temperature was set at 34 ℃ maintained for 4 hours after ROSC.Rabbits were sacrificed and specimens were taken at 24 hours after modeling.In recovery period cooling + LY294002 group(PATH+LY294002 group), LY294002 was injected intra-ventricularly at 20 minutes before resuscitation.Rapid neck cooling was started at the same time with CPR,and the target brain temperature was set at 34 ℃ maintained for 4 hours after ROSC.The rabbits were sacrificed and specimens were taken at 24 hours after modeling.In post-arrest therapeutic hypothermia group (PATH group), rapid neck cooling was begun after CPR for 1 hour,and the target brain temperature was set at 34 ℃ maintained for 4 hours after ROSC.The rabbits were sacrificed and specimens were taken at 24 hours after modeling.Animals were sacrificed by using overdose anesthetic drug.Western blot was used to detect the level of Akt p-Akt GSK-3β p-GSK-3β (ser9) protein, and TUNEL was used to observe apoptosis of tissues in each group.Multiple comparisons were performed with one-way analysis of variance (ANOVA).Results Compared with Sham group, Akt (Thr-308) phosphorylation (P-AKT) and P-GSK-3β levels in the brain neuron cytoplasm in 24 hours after CPR resuscitation in NT group was significantly reduced, and showed a gradual reduction trend (P<0.05);the P-AKT and P-GSK-3β levels in the brain neuron cytoplasm in 24 hours after CPR resuscitation in IATH group were significantly enhanced compared with NT group (P<0.05);the levels of these two kinds of protein at one hour after resuscitation in PATH group were significantly enhanced compared with NT group (P<0.05), but lower in IATH group.Intra-ventricularly injection of LY294002 made the effect of hypothermia lost, indicating that LY294002 inhibited the phosphorylation of Akt.Apoptosis cells were significantly reduced in IATH group and normothermia theatment group compared with PATH group and LY294002 group(P<0.05).Conclusions Neck cooling can reduce apoptosis in rabbit brain cells after recovery, and the protective effect on brain is best in intra-arrest therapeutic hypothermia group.LY294002 specifically block the PI3K/Akt pathway, and the protective effect of cooling on the brain can be abolished,indicating hypothermia protects the neurological function via activation of PI3K/Akt pathway.Neck cooling protects the neurological function by activating PI3K/Akt/GSK-3β, promoting the Akt activation, and increasing the expression of P-GSK3β.Specific Akt inhibitor LY294002 inhibits Akt phosphorylation of brain tissue recovery and further inhibit the phosphorylation of GSK-3β, thus abolishing protective effect of cooling on neurological function.

OBJECTIVE: To analyze the association between genetic polymorphisms of DNA repair genes of XPD (751 Lys/Gln), XPC (PAT)and susceptibility to laryngeal carcinoma. To explore the effect between DNA repair genes of XPD (751 Lys/Gln), XPC (PAT) and carcinogenesis of LSCC(laryngeal squamous cell carcinoma). METHOD: A case-control study was conducted involving 233 LSCC patients and 102 healthy controls to investigate the association between polymorphisms of XPD(751 Lys/Gln), XPC (PAT) and LSCC. All blood samples of the Han people from the Guang Dong Zone was analysze with methods of PCR, PCR-RFLP, ASA and the technique of checking DNA sequencing with sequenator. We explored the association between polymorphisms and the clinical pathologic characteristic of LSCC. The data was compute with SPSS13.0. Odds Ratios (ORs) with 95% CI for relevancy intensity were calculated using binary logistic regression analysis. REULT: There is no difference of the frequency of XPC-PAT and XPD (751 Lys/Gln) genotype between in LSCC and in healthy contradistinguish (P > 0.05). CONCLUSION There may be no association between the susceptibility to laryngeal carcinoma and the genotype of XPC-PAT and XPD (751 Lys/Gln).
Carcinoma, Squamous Cell ; genetics ; Case-Control Studies ; DNA Repair ; genetics ; DNA-Binding Proteins ; genetics ; Female ; Genotype ; Humans ; Laryngeal Neoplasms ; genetics ; Male ; Polymorphism, Genetic ; Sequence Analysis, DNA ; Xeroderma Pigmentosum Group D Protein ; genetics

Objective To investigate the expression of peroxisome proliferator-activated receptor (PPAR?)in lupus nephritis(LN)patients and the possible mechanisms of PPAR?in the pathogenesis of LN. Method PPAR?expression was examined in 21 LN patients and 5 normal kidney biopsy specimens by im- munohistochemical method.The relationship between PPAR?expression and renal pathologic changes was an- alyzed.Results Glomerular and tubular positive staining of PPAR?in LN patients was markedly up-regulated compared with that in normal kidney specimens.The distribution and expression of PPAR?in classⅣwas sig- nificantly increased compared with that in classⅤandⅡ.The relevance assay showed that there was positive relationship between active index and glomerular PPAR?immunohistochemistry staining cell numbers(r=0.94, P＜0.01 ).Conclusion This study demonstrates in vivo that PPAR?expression is increased in active LN pa- tients with pathological active inflammation.These data suggest that the increase of PPAR?expression in renal cells may play an important role in the pathogenesis of LN.

Objective To study the effect of β-amyloid peptide (Aβ1-42) on expressions of synaptophysin (Syn), dynamin Ⅰ (Dyn Ⅰ) and adaptor protein 180 (AP180) in human neuroblastoma SH-SY5Y cells. Methods Human SH-SY5Y neuroblastoma cells were treated with 0.01, 0.1, 1, 2 and 5 μmol/L Aβ1-42, and control cells were given no treatment. MTT [3-(4,5-dimethylthiazol- 2- yl) - 2, 5-diphenyltetrazolium bromide]reduction of the cells was measured by spectrophotometry. The protein levels of Syn, Dyn Ⅰ and AP180 in SH-SY5Y cells treated with 0.5 and 1 μmol/L Aβ1-42 were surveyed by Western blotting. The mRNA levels of Syn, Dyn Ⅰ and AP180 were detected by Real-time PCR in SH-SY5Y cells treated with 1 μmol/L Aβ1-42. Results SH-SY5Y cells showed obviously decreased reduction rates of MTT after exposure to Aβ1-42(0.1 μmol/L) as compared with the controls (P＜0.05), and dose-dependent negative correlation was noted in these SH-SY5Y cells. The protein level of Dyn Ⅰ in cells treated with 0.5 μmol/L Aβ1-42 was significantly decreased as compared with that in controls (P＜0.05). The protein and mRNA levels of Syn and Dyn Ⅰ in cells treated with 1 μ mol/L Aβ1-42 Were obviously decreased as compared with those in controls (P＜0.05), but the levels of AP180 were not changed. Conclusion Aβ1-42 reduces the levels of Syn and Dyn Ⅰ in SH-SY5Y cells, which might be a mechanism in eonnection with cognitive deficit of AD.

Chinese traditional patent medicine for promoting blood circulation and removing blood stasis(PBCRBS) originated from traditional Chinese medicine theory and had approved efficacy and safety standards. However, its compatibility regularity and anti-thrombotic mechanism is not clear. To analyze the compatibility regularity and anti-thrombotic mechanism of Chinese traditional patent medicine for PBCRBS, a statistical and bioinformatics analysis was carried out using traditional Chinese medicine inheritance support system (TICMISS, V2.0) and ingenuity pathway analysis (IPA). The compatibility regularity analysis shows that the most commonly used herb combinations are Danshen (Salvia miltiorrhiza Bge.), Chuanxiong (Ligusticum chuanxiong Hort.) and Honghua (Carthamustinctorius L.). The anti-thrombotic mechanism analysis reveals that 25 ingredients have an effect on 29 thrombosis related molecules which 23 molecules are related to inflammation response. Furthermore, there are 5 inflammation molecules (NOS2, PTGS2, IL6, TNF, IL1β) served as major targets. At the same time, Danshen, Chuangxiong and Honghua mainly used as sovereign herb or minister herb in the application of cardiovascular and cerebrovascular diseases. Therefore, Chinese traditional patent medicine for PBCRBS probably has an effect on anti-thrombotic activity through inhibiting the inflammatory response. In summary, the most commonly used herb combinations of Chinese traditional patent medicine for PBCRBS are Danshen, Chuanxiong and Honghua. Inhibiting inflammatory response, especially inflammation related molecules (NOS2, PTGS2, IL6, TNF and IL1β), is probably a new starting point to clarify the anti-thrombotic mechanism of Chinese patent medicine for PBCRBS.
Anti-Inflammatory Agents ; pharmacology ; Carthamus tinctorius ; Computational Biology ; Drugs, Chinese Herbal ; pharmacology ; Fibrinolytic Agents ; pharmacology ; Humans ; Inflammation ; drug therapy ; Medicine, Chinese Traditional

This study aims to investigate the genetic characteristics of BZLF1 gene and its promoter Zp of the epidemic strains in children with primary Epstein-Barr virus (EBV)-associated diseases. Total DNA was extracted from the peripheral blood of 134 children with EBV-associated infectious mononucleosis (EBV-IM) and 32 children with EBV-associated hemophagocytic lymphohistiocytosis (EBV-HLH) who were admitted to Beijing Children's Hospital from 2006 to 2011. The EBNA3C, BZLF1, and Zp genes were amplified by PCR assay. Typing of EBV was performed according to the size of the amplification product of EBNA3C gene; the amplification products of BZLF1 and Zp genes were subjected to direct sequencing, and sequence analysis was performed using BioEdit 7. 0. 9. The results were as follows: (1) EBV-1 was present in 140 samples (97.2%, 140/144) and EBV-II in 4 samples (2.8%, 4/144). (2) Three BZLF1 genotypes and their 12 subtypes (including 6 newly found subtypes) were detected in this study; there were no significant differences in the frequencies of BZLF1-A and BZLF1-B between the children with EBV-IM and EBV-HLH (P = 0.083); BZLF1-A1 was the dominant genotype in children with EBV-associated diseases; t BZLF1-A mostly had three 29-bp repeats in the first intron of BZLF1 gene, and BZLF1-B mostly had 30-bp repeats (P = 0.000), with the number of repeats varying from 1 to 13. (3) Four Zp genotypes were detected in this study, including Zp-P, Zp-V3, Zp-V4, and Zp-V1; there were no significant differences in the frequencies of these Zp genotypes between children with EBV-IM and EBV-HLH (P = 0.272, 0.252, 1.0, and 1.0, respectively). (4) The linkage analysis of BZLF1 gene and its promoter Zp showed that BZLF1-A1 was highly associated with Zp-V3 (P = 0.000), while BZLF1-B4 with Zp-P (P = 0.000); EBV-I + BZLF1 A1 was highly associated with Zp-V3 (P = 0.000), while EBV-I+BZLF1-B4 with Zp-P (P = 0.000). The conclusions are as follows: (1) BZLF1-A1 is the dominant genotype in children with EBV-associated diseases; there are mostly 29-bp repeats in the first intron of BZLF1 gene for BZLF1-A genotype and 30-bp repeats for BZLF1-B genotype. (2) Zp-P and Zp-V3 are dominant Zp genotypes of EBV in children, which shared similar detection rates. (3) BZLF1-A1 is highly associated with Zp-V3, while BZLF1-B4 with Zp-P; EBV-I+BZLF1-A1 is highly associated with Zp-V3, while EBV-I+BZLF1-B4 with Zp-P.
Child ; Child, Preschool ; China ; epidemiology ; Epstein-Barr Virus Infections ; epidemiology ; virology ; Female ; Genotype ; Herpesvirus 4, Human ; genetics ; physiology ; Humans ; Infant ; Infant, Newborn ; Introns ; genetics ; Male ; Promoter Regions, Genetic ; genetics ; Repetitive Sequences, Nucleic Acid ; genetics ; Trans-Activators ; genetics