Role of hippocampal REV-ERBα in postoperative cognitive dysfunction in rats / 中华麻醉学杂志

Objective:To evaluate the role of hippocampal REV-ERBα in postoperative cognitive dysfunction in rats.Methods:Thirty-two SPF healthy male Sprague-Dawley rats, aged 12-14 weeks, weighing 360-380 g, were divided into 4 groups ( n=8 each) using a random number table method: control group (group C), surgery group (group S), surgery + dimethyl sulfoxide (DMSO) group (group SD), and surgery + SR9009 group (group SS). Exploratory laparotomy was performed under sevoflurane anesthesia in S, SD and SS groups.Normal saline containing 0.1% DMSO was injected into hippocampal CA1 area at 1 h before laparotomy, with 2 μl on each side in group SD, and REV-ERBα agonist SR9009 (in normal saline containing 0.1% DMSO) was injected into hippocampal CA1 area at 1 h before laparotomy, with 2 μl on each side in group S+ SR9009.Morris water maze test was performed at 1 and 3 days after operation.Rats were sacrificed at 1 h after the end of Morris water maze test on day 3 after surgery, and the hippocampal tissues were obtained for determination of the expression of REV-ERBα, Brain and Muscle ARNT-Like 1 (BMAL1) protein, synaptophysin (SYN), postsynaptic density (PSD)-95 protein and N-methyl-D-aspartate receptor 2B subunit (GRIN2B) (by Western blot) and microscopic examination of the morphology of hippocampal neurons and Nissl bodies (by Nissl staining), and the viable neurons were counted. Results:Compared with group C, the percentage of time of staying at the target quadrant was significantly decreased, and the number of crossing platform was reduced on days 1 and 3 after exploratory laparotomy, the expression of REV-ERBα, BMAL1, PSD95, SYN and GRIN2B was down-regulated, and the number of viable neurons was decreased in group S and group SD ( P<0.05). Compared with group S and group SD, the percentage of time of staying at the target quadrant and the number of crossing platform were significantly increased on days 1 and 3 after exploratory laparotomy, the expression of REV-ERBα and PSD95 was up-regulated, the number of viable neurons was increased ( P<0.05), and no significant change was found in the expression of BMAL1, SYN and GRIN2B in group SS ( P>0.05). There was no significant difference in the indexes mentioned above between group S and group SD ( P>0.05). Conclusions:Activation of REV-ERBα can improve postoperative cognitive dysfunction, and the mechanism may be related to up-regulation of PSD95 expression in hippocampus and reduction of neuronal damage in rats.

Beta-sitosterol alleviates liver fibrosis in mice by blocking tprl-smad2/3 and TNF-a-nf-icb signaling pathways / 中国药理学通报

Aim To explore the effect of beta-sitosterol (BS) on liver fibrosis induced by CCL4 in mice and the mechanisms. Methods Fifty C57BL/6 male mice were randomly divided into five groups; control group (CG) , carbon tetrachloride group (CTG), low/medium/high dose of BS group ( BS-L/M/H), with 10 mice in each group. The model of hepatic fibrosis was established by injecting CCL4 in peritoneal cavity, the study lasted 30 days, and different doses of BS were given from 1st day to 15 th day. All mice were sacrificed for the observation of morphological changes and the measurement of liver index. Liver collagenous fibers were observed by HE and Masson staining, the changes of serum ( ALT and AST) were assessed by Elisa, the expressions of a-SMA and Collagen I were detected by Western blot and immunohistochemistry, and the changes of TpRl-Smad2/3 and TNF-a-NF∗kB were detected by Elisa and Western blot. Results Compared to control group, different doses of BS markedly inhibited the increase of liver index, A .T, AST, a-SMA and Collagen I in a dose-dependent n an-ner ( P < 0. 05 or P < 0. 01 ). Liver morphology, inflammatory cell infiltration and collagenous fiber irj BS groups were better than those in CCL4 group, meanwhile BS-M decreased the expression of TgKl, Smad2/3, TNF-a and p-NF-KB (P <0. 01). Conclusions BS dose-dependently inhibits mouse liver f bro-sis induced by CCL4, and its mechanism may be related to inhibiting TpRl-Smad2/3 and TNF-a-N •-kB signaling pathways.

Effect of acute sleep deprivation on learning and memory and hippocampal circadian gene expression after sevoflurane anesthesia in rats / 中华行为医学与脑科学杂志

Objective@#To investigate the role of clock gene Bmal1, Per2 and Egr1 expression in learning and memory undergoing sevoflurane anesthesia after acute sleep deprivation.@*Methods@#72 male SD rats were equally divided into four groups using a random number table (n=18) : normal control group (group Control), do not do any processing; sleep deprivation group (group SD), acute sleep deprivation for 96 h; sevoflurane group (group Sev), suffering 2.5% sevoflurane for 3 h; sleep deprivation+sevoflurane group (group SD+Sev), 96 h sleep deprivation followed by 3 h 2.5% sevoflurane inhalation. The Morris water maze, for spatial memory acquisition test, was used to measure the time percent of target quadrant and numbers of platform-site crossovers before sleep deprivation (T0) and at 1 d (T1), 3 d (T2), 7 d (T3) after inhalation anesthesia. Rats were sacrificed after spatial memory acquisition test. Brain hippocampus samples were obtained for determination of Bmal1, Per2 and Egr1 expression by Western blot, and neuron morphology was observed by the Nissl staining.@*Results@#Compared with Control group, the percentage of time in target quadrant and the numbers of platform-site crossovers were significantly decreased at T1 and T2 in Sev group (P<0.05); and compared with Control group, the percentage of time in target quadrant and the numbers of platform-site crossovers were also significantly decreased at T1, T2 and T3 in SD group rats (P<0.05). Compared with Sev group rats (the percentages of time in target quadrant: T1: (32.37±1.36)%; T2: (30.91±1.26)%; T3: (33.78±2.20)%; the numbers of platform-site crossovers: T1: (4.55±0.39); T2: (3.11±0.37); T3: (3.95±0.34)), the percentages of time in target quadrant (T1: (27.20±1.42)%; T2: (28.19±1.04)%; T3: (30.06±1.22)%) and the numbers of platform-site crossovers (T1: (3.11±0.46); T2: (3.30±0.38); T3: ( 3.20±0.39)) in SD+Sev group rats were significantly decreased at T1, T2 and T3 (all P<0.05). Compared with control group, the levels of hippocampal proteins Bmal1, Per2 and Egr1 were significantly reduced at T1 in Sev group (P<0.05). Compared with control group, the level of hippocampal protein Per2 was significantly increased, but the levels of hippocampal proteins Bmal1 and Egr1 were significantly decreased at T1 and T2 in SD group (P<0.01). Compared with Sev group, the levels of hippocampal proteins Bmal1 and Egr1 were significantly reduced at T1, T2 and T3 in SD+Sev group (P<0.01), and the protein level of hippocampal Per2 was significantly decreased at T1, but then increased at T2 and T3 in SD+Sev group (P<0.01). The hippocampal Nissl staining in CA1 at T2 revealed that there were irregular distribution of pyramidal neurons existed in Sev group, and vacuolar degeneration with vague outlines of pyramidal neurons in SD group, while pyramidal neuron atrophy and few number of Nissl bodies, compared with control group, were observed in SD+Sev group.@*Conclusion@#Acute sleep deprivation following with sevoflurane anesthesia resulted in hippocampal memory impairment, which was associated with abnormal expression of hippocampal Bmal1, Per2 and Egr1.

The inhibitory effect of Euonymus alatus against mouse hepatic fibrosis induced by CCl4 / 中国药理学通报

Aim To explore the inhibitory effect of Euonymus alatus on hepatic fibrosis induced by carbon tetrachloride (CCl4) in mice and its mechanism. Methods Eighty C57BL/6 male mice were randomly divided into eight groups: normal group, CCl4model group, Euonymus alatus(EA) ethanol extracts groups in early stage(EAE), EA ethanol extracts groups in later stage(EAL),two drug groups with low/medium/high dose(EAE-L/M/H, EAL-L/M/H), with 10 mice in each group. Fibrosis model was established by injecting CCl4in peritoneal cavity,and the study lasted for 30 days. Different doses of drugs were given from 1 st day to 15 th day in EAE while from 16 th day to 30 th day in EAL,then all mice were sacrificed to for the observation of the morphological changes and collage-nous fiber by HE and Masson staining. Liver index, ALT,AST and TNF-α were tested by ELISA. The ex-pressions of α-SMA and CollagenⅠwere measured by immunohistochemistry and Western blot. Results Compared to normal group, liver index, ALT, AST, TNF-α, α-SMA and CollagenⅠ in EA groups were lower than those in model group in a dose-dependent manner(P<0.01 or P<0.05). Liver morphology and collagenous fiber in EAE and EAL were better than those in model group in a dose-dependent manner. The effect of EAE were superior to that of the EAL in HE, Masson, α-SMA, Collagen Ⅰ indexes(P <0.05). Conclusions Euonymus alatus may inhibit the process of hepatic fibrosis in mice with dose-effect de-pendence, and drug treatment in early stage performs better,which may be related to the decrease of TNF-α that affects the expression of α-SMA and Collagen Ⅰ.

Content Determination of 8 Metal Elements in Propylgaclate and Sodium Chloride Injection by Microwave Digestion-inductively Coupled Plasma Mass Spectrometry / 中国药房

OBJECTIVE:To establish a method for the content determination of 8 metal elements in Propylgaclate and sodium chloride injection. METHODS:Microwave digestion-inductively coupled plasma mass spectrometry(ICP-MS) was adopted. Radiofrequency power was 1 530 W;cooling temperature was 4 ℃;collision gas was He gas;carrier gas was argon;flow rate of carrier gas was 1.08 L/min;integration time was 0.3 s;plasma gas flow rate was 15 L/min;the vacuum degree of quadrupole was 3.04×10－4 Pa;sampling cone aperture was 1.0 mm;interception cone aperture was 0.4 mm;the speed of sampling was 0.3 rps;data collection was repeated for 3 times. The microwave digestion power is 1 600 W,and the heating process is heated to 160℃at room temperature for 30 min,and maintained at 5 min,and then heated to 190 ℃ at a temperature of 5 ℃/min and maintained 45 min. RESULTS:The linear range of Mg and Al were 1-250 ng/mL;the linear range of Cr,Mn,Fe,Cu,Zn,Cd were 1-100 ng/ mL(all r≥0.999 0). The limits of detection were 0.063 6-1.785 0 ng/mL. RSDs of precision,stability and reproducibility tests were all lower than 4%. The recoveries were 89.65%-105.60%(RSD were 1.57%-3.98%,n＝9). RSDs of durablity were all lower than 12%. CONCLUSIONS:The method is simple,accurate,precise,stable,reproducible and durable. It can be used for content determination of 8 metal elements in Propylgaclate and sodium chloride injection.

Relationship between unintentional injury and health risk behaviors among adolescents / 中华行为医学与脑科学杂志

Objective To identify the effect of health risk behaviors on accidental injury of adoles-cents.Then analyze the strength and approach of the health risk behaviors. Methods Cluster sampling on various stages and strata methods was adopted. The adolescents from grade one to three in junior school students and grade one to three in senior high school students completed the questionaire. A total of 4408 students were surveyed and 4309 samples were valid. Path analysis was used to understand how factors could influence injury incidence. Results The incidence of adolescent injury was 15.53%. Statistical differences were found between genders (male 17.77%,female 13.36%,χ2=15.91, P<0.01),education levels (junior high school 17.36%,senior high school 13.99%,χ2=11.45, P<0.01) and cities ( Beijing 12.97%,Zhongs-han 17.95%,χ2=20.33, P<0.01) . Those who got injured were found with more health risk behaviors. Health risk behavior was the direct influencing factor to injury,as well as a possible risk factor to multiple injuries. Conclusion Health risk behaviors can strongly affect adolescent injury,and can be an indicator. Reducing health risk behaviors is important in adolescent injury prevention.

Comparison of Total DNA Extraction Methods for Studying Endophyte Diversity in Mulberry / 广州中医药大学学报

Objective To compare the advantages and disadvantages of the four DNA extraction methods according to the endophytic diversity in the roots, stems, and leaves of mulberry analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) , and by taking the yield, purity and PCR amplification as indexes. Methods Four common methods, i.e., cetyltrimethyl ammonium bromide ( CTAB) , sterile phosphate buffered saline (SPBS) vibration, liquid nitrogen grinding (LNG) , and KIT methods, were used to extract the total DNA from different tissues of mulberry, and then were compared based on the diversity analysis results for endophyte by PCR-DGGE. Results From the roots and stems of mulberry, we got the highest concentration of DNA by LNG extraction method, and got the lowest concentration by SPBS extraction method. But for the leaves of mulberry, the results of the four extraction methods were completely opposite to those for the roots and stems. For different tissues of mulberry, the purity of DNA extracted by KIT method was the best. According to the endophytic bacteria diversity analyzed by 16S rDNA PCR-DGGE, the appropriate method for extraction of DNA was LNG or CTAB, but was not KIT. And according to the results of endophytic fungi diversity analyzed by ITS PCR-DGGE, the best extraction method was KIT, and the unsuited methods varied from the tissues of mulberry. Conclusion The optimum DNA extraction method for mulberry varies from the tissues of mulberry and endophtic bacteria.

Study on transfection of adeno associated virus 2-ND4 gene into mitochondria / 中华实验眼科杂志

Background Leber hereditary optic neuropathy (LHON) is mitochondrial DNA (mtDNA) disease and mainly leads to optical nerve degeneration.Its primary mechanism is synthesis disorder of DN4 protein due to variation of mtDNA 11778 locus.So to construct a vector with exogenous normal ND4 and transfect into mitochondria is a key of gene therapy for LHON.Objective This study was to investigate the in vitro transfection of adeno-associated virus (AAV)-ND4 gene into mitochondria.Methods Human renal epithelial cell lines transfected adenovirus E1A (293 cells) were regularly cultured and divided into two groups.Framework plasmids of recombinant AAV-ND4 or simple AAV2 were added to the cell medium respectively.The expression of ND4 in cells were located 12,24,36 and 48 hours after transfected by Y03 dual fluorescent quantum dots staining.The positive response for ND4 showed the green fluorescence.Results Cultured 293 cells grew well with 80％ confluence.Abundant green fluorescence particles were seen in cytoplasm in the AAV-ND4 transfected group,but only red fluorescence from mitochondrial protein was seen in the simple AAV transfected group under the fluorescence microscope.Conclusions Exogenous ND4 protein can been successfully transfected into mitochondria using the ND4 gene constructed AAV.This result provides experimental evidence for the further study on gene therapy of LHON.

The Catechol-O-Methyltransferase Val158Met Polymorphism Contributes to the Risk of Breast Cancer in the Chinese Population: An Updated Meta-Analysis / 한국유방암학회지

PURPOSE: Catechol-O-methyltransferase (COMT) enzyme plays a central role in estrogen-induced carcinogenesis. Emerging evidence from association studies has revealed that the functional Val158Met polymorphism (rs4680 G>A) of the Catechol-O-methyltransferase gene (COMT) has been implicated in susceptibility to breast cancer in the Chinese population, while results of individual published studies remain inconclusive and inconsistent. To assess this association in the Chinese population, a meta-analysis was performed. METHODS: Eligible studies were searched on MEDLINE, Embase, Cochrane Library, China National Knowledge Infrastructure, and the Chinese Biomedicine Database. Odds ratios (ORs) with their corresponding 95% confidence intervals (CIs) were pooled to assess the association between COMT polymorphisms and the risk of breast cancer using RevMan 5.2 and Stata 12.0 software. RESULTS: The meta-analysis included 14 eligible studies, with a total of 4,626 breast cancer cases and 5,637 controls. Overall, the COMT Val158Met polymorphism (rs4680 G>A) was significantly associated with an increased risk of breast cancer in several genetic models (A/A vs. G/G: OR, 1.59, 95% CI, 1.12-2.27; A/A vs. G/A+G/G: OR, 1.62, 95% CI, 1.14-2.29; A vs. G: OR, 1.15, 95% CI, 1.00-1.32), and a subgroup analysis according to menopausal status showed that this association was especially evident among premenopausal Chinese women (A/A vs. G/G: OR, 1.87, 95% CI, 0.99-3.54; A/A vs. G/A+G/G: OR, 1.94, 95% CI, 1.03-3.63). CONCLUSION: The results of this meta-analysis indicated that COMT Val158Met variants contribute to breast cancer susceptibility in the Chinese population, particularly among premenopausal women.

Estrogendecreasingmatrixmetalloproteinase-9geneexpressioninmicewithexperimental autoimmune encephalomyelitis / 中华神经科杂志

Objective To study the regulation effect of estrogen in expression of matrix metalloproteinase-9 (MMP-9) in the central nervous system (CNS) in mice with experimental autoimmune encephalomyelitis ( EAE).Methods The 60 mice were overiectomized and 2 weeks later EAE was induced with MOG35-55 peptide in these mice.They were divided into a treatment group and a control group.The treatment group was treated with estrogen and the control group was given PBS.Clinical symptoms in these two groups were scored and compared.HE staining was used to observe inflammation in the brain and spinal cord.The MMP-9 expression in the CNS was examined by quantitative real-time PCR and immunofluorescence staining.Results The incidence of disease was lower (treatment and control group were 8/30 and 28/30 respectively) and clinical symptoms were milder (treatment and control group were 3.23±0.83 and 1.62 ±1.00 respectively,t=3.811 and P＜0.05) in the treatment group than those in the control group.HE staining showed the decreased infiltration of inflammatory cell in the treatment group (Treatment group:inflammatory score were 0.895 ±0.206,0.752 ±0.302,0.732 ±0.183 in acute,relief and chronic phase respectively;Control group:inflammatory score were 3.472 ±0.635,2.881 ±0.662,1.891 ± 0.482 in acute,relief and chronic phase respectively.t = 8.622,6.543 and 5.027,all P ＜ 0.05).The quantitative real-time PCR and immunofluorescence staining showed that the expression of MMP9 in the CNS was decreased in the treatment group.Conclusion Estrogen may decrease MMP-9 expression in the CNS,reduce inflammation and clinical symptoms in mice with EAE.

Effect and mechanism of recombinant hirudin on fibrinolysis / 药学学报

<p><b>AIM</b>To study the effect of recombinant hirudin (rH) on tPA-induced fibrinolysis and the possible mechanism of its action.</p><p><b>METHODS</b>The effect of rH on thrombin-fibrin complex (Th-Fn) was detected by 99mTc labeled rH. In the in vitro clot lysis, tPA as plasminogen activator, and recalcified plasma as plasminogen resource were used to study the influence of rH on fibrinolysis by detecting TAFIa, D-Dimer and FXIII.</p><p><b>RESULTS</b>In a canine model of femoral artery thrombosis, a clear radioactivity strip was imaged in 30 - 60 min on a part image, and the femoral vein thrombosis developed at 30 min. rH efficiently inhibited clot regeneration. Addition of TM could inhibit clot lysis obviously, and CPI could shorten the delay of clot lysis which due to TAFIa. There was a dose-dependent relationship with TM concentration and TAFI activation. FXIII activation was inhibited by low concentration of rH ( < or = 0.2 u x mL(-1)), and the level of fibrinolysis product, D-Dimer, increased.</p><p><b>CONCLUSION</b>rH could inhibit the thrombin binding to fibrin. rH inhibited the activation of TAFI and FXIII by combining with thrombin which resulted in enhancement of thrombolysis.</p>

Effects of Dachengqi Decoction on Systemic Inflammatory Response Syndrome in Mice / 广州中医药大学学报

[Objective] To investigate the mechanism of Dachengqi Decoction (DCD) in treating systemic inflammatory response syndrome ( SIRS). [ Methods ] Mouse models induced by zymosan A were administered with DCD orally. Serum endotoxin level was detected by limulus assay and TNF-? and IL-6 levels by radioimmunoassay 6, 12 and 24 hours after medication. [Results] Serum levels of endotoxin, TNF-? and IL-6 were increased in SIRS mice. After treatment with DCD, serum endotoxin level was decreased at the 6th and 12th hour ( P

STUDIES ON ANTI-AGING EFFECT OF YANGSHOU DAN / 中药新药与临床药理

The anti-aging effect of Yang Shou Dan (YSD) was studied. It was shown that YSD could obviously increase the activity of superoxicde dismutase (SOD) in erythrocyte, decrease lipid peroxides (LPO) in serum and in liver tissue and reduce the content of lipofucin in myocardium of mice. It could also prolong the mean life span, the half lethal period and the longest life time of drosophila melanogater. It is suggested that YSD exerts an anti-aging effect to a certain extent.