Objective To investigate the effect of methylene blue(MB)on liver ischemia-reperfusion (I/R) injury in rabbits.Methods Twenty-four healthy New Zealand white rabbits of both sexes welshins 2.0-2.3 kg were randomly divided into 3 groups (n=8 each):group Ⅰ sham operation(group s);group Ⅱ I/R and group Ⅲ methylene blue (group MB).The animals were anesthetized with intravenous 2% pentobarbital 30 mg/kg.Liver I/R was produced by occlusion of hepatic blood flow for 40 min followed by 60 min repeffusion.In group MB methylene blue 5 mg/kg was injected iv at 20 min before liver ischemia.Femoral artery was carmulated for MAP monitoring and blood sampling.MAP and HR were recorded immediately before(T1,baseline)and at 20 and 40 min of ischemia (T2,3) and 1,5,30,60 min(T4-7)of repedusion.Blood samples were collected at T1,T5,T6 and T7 for measurement of seruln TNF-α and IL-6 concentrations.Plasma AST and ALT activities were measured at T1,T6 and T7.Liver specimens were obtained at the end of experiment for determination of SOD activity and MDA content.Results In group I/R MAP was significantly decreased at T4-7 during reperfusion and HR at T7 as compared with the baseline at T1;while in group MB no significant change in MAP and HR Was observed during ischemia and reperfusion as compared with the baseline.The gerum TNF-α and IL-6 concentrations and the plasma ALT and AST activities were significantly increased during reperfusion as compared with the baseline immediately before ischemia in group I/R and MB and were significantly lower in group MB than in group. I/R. The SOD activity was significantly higher while MDA content was significantly lower in group MB than in group I/R. Microscopic examination showed that liver damage was less severe in group MB than in group I/R. Conclusion The administration of MB can maintain hemodynamic stability and attenuate liver I/R injury in rabbits.

Objective To investigate the role of Notch signaling pathway in cerebral ischemic tolenmce induced by clectroacupuncture (EA) preconditioning.Methods Fifty-two adult male SD rats weighing 280-320 g were randomly divided into 2 groups(n=each):control group(group C)and electroacupuncture preconditionig group(group EA).Group C received no treatment.Group EA received EA at the Baihui acupoint (GV20) for 30 min a day for 5 days.Twenty-four hours after the last preconditionig,focal cerebral ischemia was induced by middle cerebral artery occlusion (MCAO) for 120 min,followed by 72 h of reperfusion.Notch intracellular domain(NICD)expression was determined by Western blot and expression of Notch1,Notch4,Jag1,and Hes1 mRNA by real-time PCR immediately before iachemia and 24 and 72 h of reperfusion.The neurological deficit was scored at 72 h of reperfusion.The infarct volumes were then determined after evaluation of the neurological deficit score .Results There was no significant difference in Hesl mRNA and NICD expression immediately before ischemia between group EA and C(P>O.05).NICD expression was up-regulated at 24 and 72 h of reperfusion in both groups, and Hesl mRNA expression at 72 h of reperfusion in group C and at 24 h of reperfusion in group EA was up-regulated compared with those immediately before ischemia (P < 0.05). Hes1 mRNA and NICD expression was up-regulated in group C, while down-regulated in group EA at 72 h of reperfusion compared with those at 24 h of reperfusion ( P < 0.05 ). Compared with group C, the expression of Notchl,Notch4 and Jag1 mRNA was up-regulated immediately before ischemia, and Hes1 mRNA and NICD expression was up-regulated at 24 h of reperfusion while down-regulated at 72 h of reperfusion in group EA( P < 0.05). EA preconditioning significantly reduced infarct volumes and increased neurological deficit scores at 72 h of reperfusion (P < 0.05 ). Conclusion Notch signaling pathway may participate in cerebral ischemic tolerance induced by EA preconditioning.

OBJECTIVETo investigate whether pretreatment with repeated electroacupuncture (EA) at the Baihui acupoint could induce ischemic tolerance against transient focal cerebral ischemic injury in rats.

METHODSThirty male Sprague-Dawley (SD) rats were randomly divided into 3 groups (n=10 for each): the control group consisted of animals receiving no treatment, the isoflurane (ISO) group had animals that inhaled 1.5% isoflurane for 30 min a day for 5 days, and animals in the EA group received electroacupuncture at the Baihui acupoint for 30 min a day for 5 days under 1.5% isoflurane anesthesia. Twenty-four hours after the last treatment, the middle cerebral artery was occluded with No. 3 nylon monofilament for 120 min. The neurological outcomes were evaluated 24 h after reperfusion. The infarct volumes were then assessed using 2% triphenyltetrazolium chloride staining after the neurological outcome evaluation.

RESULTSThe neurological deficit score (NDS) of the EA group [1 (0-2)] was lower than that of the ISO group [2 (1-3)] and the control group [2 (1-4)], P < 0.05. The infarct volume of the EA group (38.3 +/- 25.4 mm(3)) was significantly smaller than that of the control group (220.5 +/- 66.0 mm(3)) and the ISO group (168.6 +/- 57.6 mm(3)) 24 h after reperfusion.

CONCLUSIONElectroacupuncture at the Baihui acupoint 30 min a day for 5 days significantly reduces neurological injury induced by transient middle cerebral artery occlusion.

Animals ; Electroacupuncture ; Ischemic Attack, Transient ; therapy ; Male ; Rats ; Rats, Sprague-Dawley

In this study, in order to overcome the shortcomings of the current methods used to identify Bifidobacterium animalis, such as long time, complicated operation and low adaptability of experimental environment, specific primer probes were designed based on ERIC-PCR technology to identify and detect B.animalis.Based on the genomic DNA of B.animalis HP-B1124, the ERIC-PCR reaction conditions of B.animalis HP-B1124 were optimized, and the ERIC-PCR fragments were obtained one by one and sequenced.Two pairs of specific primer probes were designed.The accuracy, specificity, limitation and universality of the two pairs of primer probes were evaluated, and the two pairs of specific primer probes were used for testing the products containing B.animalis in the commercially published formula.The two pairs of specific primer probes designed in this study could be used for identified strains of B.animalis more simply, quickly and targeted.This method has optimized the current relatively traditional methods of pure culture and plate counting identification of B.animalis, and has solved the high requirements of SNP genotyping technology and real-time fluorescence quantitative PCR method for experimental equipment and reagents in the identification of B.animalis to a certain extent.It has the characteristics of low cost, high specificity and earn a broad market development prospect.

Flavonoids have attracted much attention due to their good anti-inflammatory, anti-oxidation and anti-tumor effects. At present, the extraction of flavonoids is mainly based on organic solvent, while the researches on the use of green and safe solvents are quite limited. Therefore, in the present study, different types of deep eutectic solvents (DESs) were applied to investigate their effect on extraction of flavonoids and optimize the process, also investigate the recovery efficiency of DESs and evaluate the recovery method for total flavonoids. The extraction yield of the total flavonoids acted as the comprehensive evaluation indexes, and a central composite design (CCD) of response surface methodology (RSM) was employed to further optimize the alcohol-based DES extraction conditions. The results showed that the optimized extraction conditions were as follows: water-DES ratio of 27%, solid-liquid ratio of 15 mL·g⁻¹, extraction temperature of 83 °C and extraction time of 42 min in ChCl-glycerol at 1:4 ratio. Under these conditions, the mean experimental value of the extraction yield (75.05 mg·g⁻¹) corresponded well with the predicted value (77.86 mg·g⁻¹). Moreover, these experimental results showed more advantages such as in higher efficiency, economy and environmental protection as compared with previously reported conventional extraction methods. In addition，the recovery yield of the total flavonoids from the DESs extraction solution achieved 97.88% by using AB-8 macroporous resin, and 88.12% desorption ratio can be achieved by 100% ethanol with 5 times resin content. After the above treated DESs were collected, the extraction yield with the same method reached 95.23%, indicating that the method of macroporous resin can be used for efficient and simple recovery and reuse. This study suggests that DESs can be used as a kind of sustainable and efficient natural extraction solvents for extraction of flavonoids from Prunella vulgaris.