OBJECTIVE:To establish a method for simultaneous determination of danshensu,salvianolic acid B,protocatechu-ic aldehyde,paeoniflorin and ferulic acid in Jingzhi guanxin granule. METHODS:HPLC was performed on the column of Zorbax Eclipse XDB-C18 with mobile phase of acetonitrile-methanol-0.5% H3PO4(gradient elution)at a flow rate of 1.0 ml/min,the detec-tion wavelength was 280 nm(for danshensu,protocatechuic aldehyde,salvianolic acid B),230 nm(for ferulic acid)and 320 nm (for paeoniflorin),column temperature was 30 ℃,injection volume was 10 μl. RESULTS:The linear range was 1.19-478.34 μg/ml for danshensu(r=0.999 9),0.11-44.93 μg/ml for protocatechuic aldehyde(r=0.999 9),7.49-995.20 μg/ml for salvianolic acid B (r=0.999 7),0.95-379.39 μg/ml for paeoniflorin (r=0.999 9) and 0.01-3.12 μg/ml for ferulic acid (r=0.999 5);the limits of quantitation were 1.91 ng,0.36 ng,150.00 ng,2.74 ng and 0.10 ng,limit of detection were 0.96 ng,0.10 ng,45.00 ng,1.52 ng and 0.03 ng;RSDs of precision,stability and reproducibility tests were lower than 2%;recoveries were 98.06%-99.47%(RSD=0.52%,n=6),98.01%-99.49%(RSD=0.70%,n=6),98.44%-99.45%(RSD=0.37%,n=6),96.94%-100.71%(RSD=1.27%,n=6)and 95.44%-100.44%(RSD=1.90%,n=6). CONCLUSIONS:The method is simple and accurate,and suitable for the simultaneous determination of danshensu,salvianolic acid B,protocatechuic aldehyde,paeoniflorin and ferulic acid in Jingzhi guanxin granule.

Objective To evaluate the effects of hydrogen on the expression of Rho-associated protein kinase 1(ROCK1)and mammalian diaphanous-related formin 1(mDia1)in intestinal tissues of septic mice.Methods Ninety male C57BL/6 mice,weighing 20-25 g,aged 6 weeks,were divided into 3 groups(n=30 each)using a random number table:control group(group C),sepsis group(group S)and sepsis plus hydrogen group(group SH).Sepsis was produced by cecal ligation and puncture(CLP).Group SH inhaled 2% hydrogen for 1 h starting from 1 and 6 h after CLP.Twenty mice in each group were selected and observed for 7-day survival rate.Ten mice in each group were sacrificed at 24 h after CLP,and blood samples were obtained from hearts to measure the activity of serum diamine oxidase(DAO)and to count the colony-forming units after bacterial culture.The small intestinal samples were obtained for microscopic examination of pathological changes and for determination of ROCK1 and mDia1 positive cell rates(using immunohistochemical staining)and expression of intestinal epithelial junctional protein E-cadherin(by immunofluorescent staining).Intestinal damage was assessed and scored.The ratio of ROCK1 to mDia1 positive cell rates(ROCK1/mDia1 ratio)was calculated.Results Compared with group C,the survival rate was significantly decreased,the serum DAO activity,colony counts and intestinal damage scores were increased,ROCK1 and mDia1 positive cell rates were increased,and the expression of E-cadherin was down-regulated in S and SH groups,ROCK1/mDia1 ratio was increased in group S(P<0.05),and no significant change was found in ROCK1/mDia1 ratio in group SH(P>0.05).Compared with group S,the survival rate was significantly increased,the serum DAO activity,colony counts and intestinal damage scores were decreased,the ROCK1 positive cell rate was decreased,the mDia1 positive cell rate was increased,ROCK1/mDia1 ratio was decreased,and the expression of E-cadherin was up-regulated in group SH(P<0.05).Conclusion The mechanism by which hydrogen improves intestinal barrier function may be related to down-regulation of ROCK1 expression,up-regulation of mDia1 expression and correction of the imbalance in ROCK1/mDia1 ratio in intestinal tissues of septic mice.

Objective To investigate the relationship between the mechanism underlying hydrogeninduced improvement of intestinal barrier function and RhoA-mDia1 signaling pathway in septic mice.Methods Sixty male C57BL/6 mice,weighing 20-25 g,aged 6-8 weeks,were divided into 3 groups (n =20 each) using a random number table method:control group (group C),sepsis group (group S),and sepsis plus H2 group (group SH).Sepsis was produced by cecal ligation and puncture (CLP) in mice anesthetized with chloral hydrate.Mice in group SH inhaled air containing 2％ hydrogen for 1 h starting from 1 and 6 h after CLP.At 24 h after CLP,blood samples were obtained by cardiac puncture to measure the activity of serum diamine oxidase (DAO) and to count colony-forming unit (CFU) after bacterial culture,the small intestinal samples were obtained for microscopic examination of the ultrastructure of intestinal epithelial cells (with a transmission electron mnicroscope) and for measurement of the expression and distribution of tight junction protein ZO-1 in intestinal epithelial cells (by immunofluorescence) and expression of RhoA,expression and phosphorylation of myosin phosphatase target subunit 1 (MYPT1) and expression of mDia1 (by Western blot).The p-MYPT1/MYPT1 ratio was calculated.Results Compared with group C,the serum DAO activity and whole blood CFU counts were significantly increased,the expression of RhoA was up-regulated,and the expression of ZO-1 was down-regulated in S and SH groups,the p-MYPT1/MYPT1 ratio was significantly increased,and the expression of mDia1 was down-regulated in group S,and the p-MYPT1/MYPT1 ratio was significantly decreased,and the expression of mDia1 was up-regulated in group SH (P＜0.05).Compared with group S,the serum DAO activity,whole blood CFU counts and p-MYPT1/MYPT1 ratio were significantly decreased,the expression of RhoA was down-regulated,the expression of ZO-1 and mDia1 was up-regulated (P＜0.05),and the ultrastructure changes of intestinal tissues were significantly improved in group SH.Conclusion The mechanism by which hydrogen improves intestinal barrier function is related to inhibiting RhoA activity and increasing mDia1 activity in intestinal epithelial cells of septic mice.

Objective:To retrospectively evaluate the effects of lung-protective ventilation strategies on postoperative pulmonary complications in the patients undergoing long-time neurosurgery.Methods:Based on the duration of anesthesia>8 h, the clinical data from patients underwent elective first craniotomy under general anesthesia from January 1, 2019 to December 31, 2021 were retrospectively collected. Patients were divided into lung-protective ventilation group (group L) and conventional mechanical ventilation group (group C) according to whether lung-protective ventilation was performed during operation. The baseline characteristics, intraoperative condition, postoperative pulmonary complications and hospitalization of patients in the two groups were recorded.Results:Compared with group C, the tidal volume was significantly decreased, positive end-expiratory pressure and respiratory rate were increased, the incidence of intraoperative hypoxemia was decreased, the number of patients with grade 1 according to the severity grade of postoperative pulmonary complications was significantly increased, and the length of hospital stay was shortened in group L ( P<0.05). There were no statistically significant differences in the baseline characteristics, incidence of postoperative pulmonary complications at 7 days after surgery and other parameters between group L and group C ( P>0.05). Conclusions:Lung-protective ventilation strategies can reduce the severity of postoperative pulmonary complications in the patients undergoing long-time neurosurgery.