0.05),and presented significant increase after half a year of retention(P

Objective To investigate the impact of tanshinone Ⅱ A on the level of brain NMDAR1 protein in rats after cardiopulmonary resuscitation and its effects of brain function protection.Methods Seventy-eight SD male rats were randomly (random number) divided into three groups:group A (sham group,n =6),group B (control group,n =36) and group C (Tanshinone Ⅱ A intervention group,n =36).All animals were induced to be models of cardiac arrest by choking.The rats of group C received intravenous injection of Tanshinone Ⅱ A in dose of 15 mg/kg immediately at initiation of resuscitation,while rats of group B were intravenous injected same amount of normal saline instead.Brains tissues of all rats were taken at 1,6,12,24,48 and 72 h after the restoration of spontaneous circulation (ROSC).Immunohistochemical staining method was applied for measuring the levels of brain tissue NMDAR1 and Caspases-3,while water content of the brain was detected by wet and dry weight ratio.The experimentaldata were analyzed by using one-way ANOVA.Results (①)The level of brain NMDAR1 protein in group B increased at 1 h and reached its peak at 6 h after ROSC,then its level gradually declined and dropped below normal at 48 h,72 h,and there were significant difference in variation of NMDAR1 protein levels in comparison with the group A (P ＜ 0.05) ; the NMDAR1 protein levels at 1,6,12 h in group C were significantly lower than those in group B at the same intervals (P ＜ 0.01),but no significant differences were seen at 24,48,72 h (P ＞ 0.05).(②)The level of brain Caspases-3 in group B increased after ROSC,and reached its peak at 48 h after ROSC,then declined and maintained above normal at 72 h,and this variation was significantly different from that of the group A (P ＜ 0.01) ; while the levels of caspase-3 at 1,6,12,24 h in group C were significantly lower than those in group B (P ＜ 0.01),but thses differences at 48,72 h were still significant (P ＜ 0.05).(③)The water content of brain tissue in group B increased at 1 h and reached its peak at 24 h after ROSC,then gradually decreased from 48 h,but maintained above normal at 72 h,and this trend of variation was significantly different from that of group A (P ＜ 0.01 or P ＜ 0.05).Compared with group B,water content of brain tissue in group C decreased more significantly (P ＜ 0.01).Conclusions Tanshinone Ⅱ A down-regulates brain NMDAR1 protein level at early stage in rats as well as significantly inhibits the level of Caspases-3 thereby ameliorating brain edema after cardiopulmonary resuscitation.

By using extracellular single unit recording technique, locally suppressive effects of a single dose of ketamine on sub-cutaneous (s. c. ) bee venom-induced increase in firing of wide dynamic-range (WDR) neurons in spinal dorsal horn were investi-gated on urcthane-chloralose anesthetized cats. Injection of bee venom s.c. into the cutaneous receptive field (RF) resulted in asingle phase of prolonged, persistently increased firing of WDR neurons over background activity for more than 1 h. Local pre-treatment with ketamine (100 mM, 0. 1 m l) into the center of RF where bee venom was injected produced a dramatic suppressionof the increased neuronal firing by 60% (3.10± 0.42 spikes/s, n= 5) when compared with saline pre-treated group (7.61 ± 0.17spikes/ s. n = 5 ). Moreover, local post-treatment with the same dose of ketamine also produced a profound suppression of the in-creased neuronal activity by 81% (1.51±0.06 spikes/s, n=5) when compared with the saline post-treated group (7.76±0.15spikes s, n=5). However, s.c. administration with the same dose of ketamine into a symmetrical region on the bee venom un-treated contralateral hindpaw produced no affection on the increased firing of the WDR neurons, suggesting that the suppressiveaction of local ketamine was not the result of systemic effects. The present result suggests that ketamine may exert its localantinociceptive effects mainly through the peripheral NMDA receptors in addition to its partially potential blocking effects onsodium and voltage-sensitive calcium channels.

OBJECTIVE:To study the general pattern and characteristics of the ADRs associated with Houttuynia cordata injection(HCI).METHODS:An analysis was conducted on55cases of ADRs induced by HCI reported in domestic medical jou_ rnals in recent years.RESULTS:The ADRs of HCI were not related to the dosage given.The onset of ADRs was commonly within30min during infusion.The ADRs mostly occurred in groups of patients under age20and from30to59years of age.The ADRs could involve multiple organs and systems,and the clinical manifestations were varying,of which allergic reaction was the commonest one including anaphylactic shock.CONCLUSION:The mechanism of allergic reaction induced by HCI remains to be further studied.Clinical physicians,nurses and pharmacists should pay more attention to the ADRs of HCI.

Objective To determine whether sulfasalazine stimulates hepatic stellate cell (HSC-T6) apoptosis and its possible mechanism. Methods CCK-8 assay, acridine orange/ethidium bromide(AO/EB) and Annexin Ⅴ FITC/PI were used to determine cell growth and cell apoptosis. The expression of NF-?B P65, phospho-IKK and phospho-I?B was detected by Western blotting. The nuclear translocation of HSC-T6 P65 was observed with laser confocal microscopy. Results Sulfasalazine displayed a strong growth inhibition and promoting apoptosis effect on HSC-T6 cells in a dose and time-dependent manner. Sulfasalazine, but not 5-aminosalicylic acid or sulfapyridine, inhibited the activation of NF-?B by down-regulating the expressions of P-IKK, P-I?B and the nuclear translocation of P65. Conclusion Sulfasalazine can inhibit NF-?B activity and promote apoptosis in HSC-T6 cells, where the Rel/NF-?B/I?B/IKK pathway plays an important role in HSC survival.

Objective To study the activity of N acetylglucosaminyltransferase Ⅲ, Ⅳ and V in extrahepatic cholangiocarcinoma and its significance.Methods The activities of N acetylglucosaminyltransferase(GnT) Ⅲ, Ⅳ and Ⅴ in extrahepatic bile duct carcinoma (EBDC) of 15 cases were determined by reverse phase HPLC (high performance liquid chromatography), and results were compared with benign biliary diseases in 15 cases. Results Compared with normal control, that the specific activities of GnT Ⅲ and GnT Ⅴ increased by 3 3 and 13 5 fold respectively in all of the EBDC samples(all P

Objective To observe the effects of arsenic trioxide(As2O3)on the expressions of TNF-?,Fas and bcl-2 in lung adenocarcinoma cells(LAC)and to explore the mechanism of arsenic trioxide inducing apoptosis.Methods The expressions of TNF-?,Fas and bcl-2 in lung adenocarcinoma cells pretreated by arsenious acid were determined by the double antibody sandwich ABC-ELISA method.Results Compared with the control group,As2O3 showed no effects on the contents of bcl-2 in lung adenocarcinoma cells after 72 hours treatment,but increased the contents of TNF-? and Fas significantly,and the effects in different concentration groups had significant differences.The protein expressions of TNF-? and Fas showed a tendency of concentration-dependent increasing.Conclusions The results suggest that As2O3 induces the apoptosis of LAC cells possibly by up-regulating the expression of TNF-? and Fas.

【Objective】 To explore the therapeutic mechanismof arsenic trioxide(AS2O3),the active component of arsenolitumininhibitingtumor.【Methods】Humanlung adenocarcinoma cells(LAC) at logarithmic growth phase were culturedin vitro for 24 hours,and then were cocultured with arsenic trioxide(AS2O3) in the dosages of 0.75,1.0,1.5 and 2.0 mg/Lrespectively.Meanwhile,model control group was set up.The morphological features of LAC in different groups were observed under reverse microscope,growth-inhibitory rate of LAC was examined by methylthiazolyltetrazolium(MTT) assay,andthe expression of p53 gene was detected by enzyme-linked immunosorbent assay(ELISA) method.【Results】AS2O3in different dosages obviously inhibited the proliferation of LACin a time-effect manner,and markedly increased the expression of p53 gene(P

E.coli single-stranded DNA-binding protein (SSB) plays an important role in replication, recombination and repair of DNA and is thus crucial for the survival of the bacteria.We described a high expression and efficient purification scheme and kinetic assay of interaction with its substrate, single-stranded DNA (ssDNA). A ssb gene (537 bp) for encoding SSB was obtained by PCR amplification from E.coli K-12 genome. The expression vector of the fusion protein SSB was constructed by attaching ssb gene to pQE30. SSB fusion protein was expressed in M15 E.coli strain induced by IPTG. SDS-PAGE analysis revealed that the expected protein with a molecular weight 20.6kDa was soluble and amounted to about 30% of the total bacterial protein. SSB protein was purified by immobilized metal (Ni2+) chelation affinity chromatography and the purity was about 90%. The resulting SSB protein was a correctly folded tetramer analyzed by gel filtration. It could bind ssDNA with equilibrium dissociation constant (KD) of 4.79×10-7 mol/L as determined by surface plasmon resonance.