Establishment and application of quality evaluation method for Xiaochaihu Granules based on calibrator samples / 中国中药杂志

With reference to the production process documented in Chinese Pharmacopoeia, this paper prepared the calibrator samples of Xiaochaihu Granules from multiple batches and established a method for fingerprint analysis and content determination that could be used to evaluate Xiaochaihu Granules available in market. Multiple batches of Chinese herbal pieces contained in Xiaochaihu Granules were collected for preparing the calibrator samples according to the process in Chinese Pharmacopoeia. Following the establishment of fingerprints for calibrator samples by UHPLC, the method for determining the contents of saikosaponin B2, saikosaponin B1, baicalin, wogonoside, baicalein, liquiritin, glycyrrhizin G2 and glycyrrhizic acid in Xiaochaihu Granules was established. The experimental results showed that the fingerprints of calibrator samples had 26 common peaks, covering the chemical compounds of main herbs Bupleuri Radix, Scutellariae Radix, Changii Radix, Glycyrrhizae Radix et Rhizoma, and Rhizoma Zingiberis Recens. The similarity of fingerprints for 47 batches of Xiaochaihu Granules from 31 companies with the calibrator sample fingerprint ranged from 0.74 to 0.99, indicating good applicability of the established fingerprint. The contents of main components baicalin, saikosaponin B2, and glycyrrhizic acid in Xiaochaihu Granules were within the ranges of 22.917-49.108 mg per bag(RSD 19%), 0.28-2.19 mg per bag(RSD 62%), and 0.897-6.541 mg per bag(RSD 41%), respectively. The quality difference in saikosaponin B2, and glycyrrhizic acid among different manufacturers was significant. The fingerprint analysis and content determination method for calibrator samples of Xiaochaihu Granules prepared according to the production process in Chinese Pharmacopoeia has been proved suitable for evaluating the quality of Xiaochaihu Granules from different manufacturers. Saikosaponin B2, glycyrrhizic acid, and liquiritin should be added as content control indicators for Xiaochaihu Granules, aiming to further improve the product quality.

Effect of two administration routes of Shenmai Injection () on pulmonary gas exchange function after tourniquet-induced ischemia-reperfusion / 中国结合医学杂志

<p><b>OBJECTIVE</b>To compare the effect between nebulized and intravenous administration of Shenmai Injection () on pulmonary gas exchange function of patients following tourniquet-induced lower limb ischemia-reperfusion.</p><p><b>METHODS</b>Thirty-eight patients scheduled for lower extremity surgery were randomized into three groups using the closed envelop method: Shenmai Injection was administered 30 min before tourniquet inflflation by nebulization [0.6 mL/kg in 10 mL normal saline (NS)] in the nebulization group or by intravenous drip (0.6 mL/kg dissolved in 250 mL of 10% glucose) in the intravenous drip group, and equal volume of NS was given intravenously in the NS group; 15 in each group. Arterial blood gases were analyzed, serum levels of malonaldehyde (MDA) and interleukine-6 (IL-6) and interleukine-8 (IL-8) were determined using the method of thiobarbituric acid reaction and enzyme-linked immuno sorbent assay respectively just before tourniquet inflflation (T0), and at 0.5 h (T1), 2 h (T2), 6 h (T3) after tourniquet deflflation.</p><p><b>RESULTS</b>Compared with baselines at T0, MDA levels signifificantly increased at T2, T3 in the NS group and at T3 in the nebulization group, and IL-6 and IL-8 levels were signifificantly increased at T2, T3 in NS, the intravenous drip and the nebulization groups (P <0.05). Arterial pressure of oxygen (PaO) at T3 was decreased, while alveolararterial oxygen tension showed difference (PA-aDO) at T3 in the NS group; RI at T3 in both intravenous drip and the nebulization groups were enhanced (P <0.05). Compared with the NS group, MDA and IL-8 levels at T2, T3, IL-6 at T3 in the intravenous drip group, and IL-8 at T3 in the nebulization group were all remarkably increased (P <0.05). Additionally, MDA level at T3 in the nebulization group was higher than that in the intravenous drip group (P <0.05).</p><p><b>CONCLUSIONS</b>Intravenous administration of Shenmai Injection provided a better protective effect than nebulization in mitigating pulmonary gas exchange dysfunction in patients following tourniquet-induced limb ischemia-reperfusion.</p>

Effect of small interfering RNA targeting caspase-12 pretreatment on lung ischemia/reperfusion injury in mice / 中华麻醉学杂志

Objective To evaluate the effect of small interfering RNA targeting caspase-12 (caspase-12-siRNA) pretreatment on lung ischemia/reperfusion (I/R) injury in mice.Methods Forty male C57BL/6J mice,aged 6-8 weeks,weighing 16-24 g,were randomly allocated into 4 groups (n =10 each) using a random number table:sham operation group (group S),group I/R,negative control group (group NC) and caspase-12-siRNA pretreatment group (group siRNA).Lung I/R was induced by clamping the left pulmonary hilum for 30 min followed by 3 h reperfusion in anesthetized mice in IR,NC and siRNA groups.At 48 h before ischemia,negative control siRNA 20 μg and caspase-12-siRNA 20 μg were instilled intranasally in NC and siRNA groups,respectively,and the total volume was 50 μl.At 3 h of reperfusion,the animals were sacrificed and the left lung was removed for determination of wet/dry lung weight (W/D) ratio and lung water content in lung tissues and for microscopic examination.Pulmonary ultrastructure was examined with electron microscope.The quantitative evaluation index (QEI) for alveolar damage and apoptosis rate were calculated.Results Compared with group S,W/D ratio,lung water content,QEI for alveolar damage and apoptosis index were significantly increased in IR and NC groups,QEI for alveolar damage and apoptosis index were increased in group siRNA (P ＜ 0.05).Compared with IR and NC groups,W/D ratio,lung water content,QEI for alveolar damage and apoptosis index were significantly decreased (P ＜ 0.05),and the pathological changes of lungs were alleviated in group siRNA.There was no significant difference in the indices mentioned above between groups IR and NC (P ＞ 0.05).Conclusion Caspase-12-siRNA pretreatment can attenuate lung I/R injury in mice.

Role of cyclooxygenase-2 in ventilator-induced lung injury in rats / 中华麻醉学杂志

Objective To evaluate the role of cyclooxygenase-2 (COX-2) in ventilator-induced lung injury in rats.Methods Thirty healthy adult male Sprague-Dawley rats,aged 3.0-3.5 months,weighing 300-350 g,were randomly divided into 3 groups (n =10 each) using a random number table:traditional tidal volume group (group T,VT =8 ml/kg),large tidal volume group (group L,VT =40 ml/kg) and NS398 (selective COX-2 inhibitor,VT =40 ml/kg) group (group N).In group N,8 mg/kg NS398 (in 2 ml of 10％ dimethyl sulfoxide) was injected intraperitoneally at 1 h before ventilation,while dimethyl sulfoxide 2 ml was administrated instead of NS398 in T and L groups.After 4 h of mechanical ventilation,arterial blood samples were obtained for blood gas analysis and PaO2 was recorded.The animals were sacrificed and lungs removed for microscopic examination and for determination of wet/dry lung weight ratio (W/D ratio) and concentrations of total protein,intercellular adhesion molecule-1 (ICAM-1),tumor necrosis factor-alpha (TNF-α),NO and 6-keto-prostaglandin F1alpha (6-keto-PGF1 α) in bronchoaveolar lavage fluid (BALF).Pulmonary permeability index (PPI) and TXB2/6-keto-PGF1 α ratio were calculated.Results Compared with group T,PaO2 was significantly decreased,W/D ratio and PPI were increased,the total protein,ICAM-1,TNF-α and NO levels and TXB2/6-keto-PGF1 α ratio in BALF were increased in group L(P ＜ 0.05).PaO2 was significantly higher,W/D ratio and PPI were lower,and total protein,ICAM-1,TNF-α and NO levels and TXB2/6-keto-PGF1 α ratio in BALF were lower in group NS than in group L (P ＜ 0.05).The damage to lung tissues was severe in group L,and obviously alleviated in group N.Conclusion COX-2 is involved in ventilator-induced lung injury in rats.

Effect of curcumin on caspase-12 and apoptosis in pulmonary ischemia/reperfusion injury mice / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the effect of curcumin (CUR) on cycteinyl aspirate specific protease-12 (Caspase-12) and pneumocyte apoptosis in pulmonary ischemia/reperfusion (I/R) injury mice.</p><p><b>METHODS</b>The in vivo unilateral in situ pulmonary I/R injury mouse model was established in C57BL/6J mice. Sixty experimental mice were randomly divided into six groups by random digit table, i. e., the sham-operation group (Sham), the I/R group, the I/R + dimethyl sulfoxide group (I/R + DMSO), the I/R + low dose CUR pre-treated group (I/R + CUR-100), the I/R + middle dose CUR pre-treated group (I/R + CUR-150), the I/R + high dose CUR pre-treated group (I/R + CUR-200), 10 in each group. Mice were euthanized and their left lungs were excised. Wet lung weight to dry lung weight (W/D) and the total lung water content (TLW) were tested. The morphological changes of the lung tissue were observed and index of quantitative evaluation for alveolar damage (IQA) detected under light microscope. The ultra-microstructure of the lung tissue was observed under electron microscope. The mRNA and protein expression levels of Caspase-12 and glucose regulated protein (GRP78) were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. Apoptosis index (AI) of the lung tissue was determined by terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) method.</p><p><b>RESULTS</b>Compared with the Sham group, expression levels of Caspase-12, GRP78 mRNA and protein all significantly increased in the I/R group (P < 0.05); W/D, TLW, IQA, and AI were all notably higher (P < 0.05, P < 0.01); the morphological and ultrastructural injury of the lung tissue were notably observed in I/R group. Compared with the I/R + DMSO group, expression levels of GRP78 mRNA and protein were increasingly higher in the I/R + CUR-100 group, the I/R + CUR-150 group, and the I/R +CUR-200 group (P < 0.05), expression levels of Caspase-12 mRNA and protein were lower (P < 0.05); W/D, TLW, IQA, and AI also decreased (P < 0.05, P < 0.01); the morphological and ultrastructural injury of the lung tissue were gradually alleviated in the I/R + CUR groups.</p><p><b>CONCLUSION</b>CUR had better effect on the lung protection against I/R injury, which might be related to inhibition for pneumocyte apoptosis associated with Caspase-12 in excessive unfolded protein response (UPR).</p>

Effects of shenmai injection on pulmonary aquaporin 1 in rats following traumatic brain injury / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Aquaporin-1 (AQP1) has involved in fluid transport in diverse pulmonary edema diseases. Our study aimed to explore the dynamic changes of AQP1 in pulmonary water metabolism in rats following traumatic brain injury (TBI) and the protective effect provided by shenmai injection.</p><p><b>METHODS</b>Sixty male Sprague Dawley rats weighting 280 - 300 g were randomly divided into three groups: the normal control group, the model group and the shenmai injection (SMI) group. One piece skull was taken away without injuring cerebral tissue in normal control group, while rats in model group and SMI group were subject to free fall injury in the cerebral hemisphere. Rats in model group received intraperitoneal normal sodium (15 ml/kg) at one hour post-injury and the same dose of shenmai injection instead in SMI group, respectively. The expression of AQP1 was detected by immunohistochemical analysis and semi-quantitative RT-PCR at 0 hour, 10 hours, 72 hours and 120 hours after TBI. Arterial blood gas analysis and lung wet to dry were also measured.</p><p><b>RESULTS</b>AQP1 was mainly presented in the capillary endothelium and slightly alveolar epithelial cells in three groups, but the expression of AQP1 in the normal control group was positive and tenuous, weakly positive in the model and SMI groups, respectively. Compared with normal control group, AQP1 mRNA levels were down regulated in the model and SMI groups at 10 hours, 72 hours and 120 hours (P < 0.05). While AQP1 mRNA levels in the SMI group was up-regulated than that in the model group (P < 0.05). Lung wet to dry weight ratio (W/D) in the model and SMI groups at 10 hours were higher than that in normal control group (P < 0.05). Compared with normal control group, PaO2 was markedly lower in the model and SMI groups (P < 0.05), but there were no statistically significant differences between model and SMI groups (P > 0.05).</p><p><b>CONCLUSIONS</b>The decreased AQP1 expression may be involved in the increased lung water content and dysfunction of pulmonary water metabolism following TBI. The treatment with SMI could improve water metabolism by promoting AQP1 expression.</p>

Protective effect of Shenmai injection on lung injury induced by cardiac pulmonary bypass / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the protective effect of Shenmai Injection (SMI) on acute lung injury (ALI) induced by cardiac pulmonary bypass (CPB).</p><p><b>METHODS</b>Thirty patients, scheduled to receive cardiac valve replacement by CPB for the first time, were equally randomized into 2 groups, the treated group and the control group. Cardiac valve replacement was performed under extracorporeal circulation after general anaesthesia. SMI 0.6 mL/kg was given to the treated group by adding in 250 mL physiological saline for intravenous dripping at the time between intubation under anaesthesia and CPB, while 250 mL physiological saline was given to the control group alone. Blood-gas analysis was performed with blood withdrawal from the radial artery to record PaO2, PaCO2, fraction of inspired oxygen (FiO2), by them the alveolar-arterial difference of partial oxygen pressure [P(A-a) DO2] was calculated, and the respiratory index (RI), the blood concentrations of soluble intercellular adhesion molelue-1 (sICAM-1), endothelin-1 (ET-1) and nitric oxide (NO) were measured at various time points, i.e. before anesthesia induction, 0.5 h, 2 h, 6 h and 24 h after ending CPB.</p><p><b>RESULTS</b>All indices wer not significantly different between the two groups before operation (P > 0.05). After CPB, P(A-a) DO2 and RI were gradually elevated and reached the peak at 2 h after ending CPB, the increment in the treated group was lower than that in the control group (P < 0.05 or P < 0.01). Compared with the P(A-a) DO2 before anesthesia induction, P (A-a) DO2 at 0.5-24 h was statistically different (P < 0.05 or P < 0.01). Compared with the RI before anesthesia induction, RI at 2-24 h was statistically different (P < 0.05 or P < 0.01). Blood concentrations of sICAM-1 gradually raised after CPB, and reached the peak at 2 h after ending CPB, showed a higher level at 0.5-6 h after ending CPB as compared with that before anesthesia induction (P < 0.05 or P < 0.01). Blood ET-1 showed a figure of increasing as sICAM did (P < 0.05 or P < 0.01), and with a lower degree at 0.5-2 h after ending CPB in the treated group (P < 0. 01). Blood NO obviously reduced after CPB, it was lower at 0.5-24 h after ending CPB than at that before anesthesia induction (P < 0.01), and the decrement was lesser in the treated group (P < 0.05 or P < 0.01).</p><p><b>CONCLUSION</b>SMI can attenuate the acute lung injury after CPB by way of inhibiting vascular endothelial cell adhesion with inflammatory cells, antagonizing lipid peroxidation, and improving the ventilation and oxygenation function of lung.</p>

Effect of Shenmai injection on expression and activity of heme oxygenase-1 in reperfusion injury after pulmonary ischemia in rabbits / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the effect of Shenmai injection the expression of heme oxygenase-1 (HO-1) in rabbits with reperfusion injury after pulmonary ischemia.</p><p><b>METHOD</b>Single lung ischemia/reperfusion injury animal model was used in vivo. Twenty rabbits were randomly divided into two groups (n = 10, in each), pulmonary ischemia and reperfusion injury (PIRI) group and I-R + Shenmai injection group. The tissue slides were stained by in situ hybridization (ISH) for HO-1 to detect the expression of HO-1 in lung and to analyze the absorbance. Wet to dry ratio of lung tissue weight (W/D) and the injured alveoli rate (IAR) were measured at 180 minutes after lung reperfusion. Meanwhile the lung tissue slide was prepared for electron microscopic observation at 180 minutes after reperfusion.</p><p><b>RESULT</b>HO-1 expression was upregulated in two groups in the pulmonary endothelial cells, part of pulmonary vascular smooth muscle cells, extima of vessels and epithelial cells of airway, the absorbance was 0.148 +/- 0.013, 0.158 +/- 0.012, respectively. The Shenmai injection group showed higher absorbance than those of the IRI group (P < 0.01), lower W/D and IAR values than those of the IRI group (P < 0.01) significantly and lighter abnormal changes of the lung tissue in morphologically than those of the PIRI group.</p><p><b>CONCLUSION</b>Shenmai injection possesses notable protective effects on PIRI in rabbits by increasing the expression of HO-1 in lung.</p>

Protective effect of ligustrazine and propofol on peri-operational liver ischemia-reperfusion injury / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the protective effect and mechanism of ligustrazine (LGT) and propofol (PRO) on peri-operational liver ischemia-reperfusion injury (HIRI).</p><p><b>METHODS</b>Thirty-six patients scheduled for hepatic surgery were randomly divided into the control group, the LGT group, the PRO group and the LGT + PRO group, 9 patients in each group. Changes of superoxide dismutase (SOD), lipid peroxide (LPO), ratio of thromboxane B2 (TXB2) and 6-keto-prostaglandin F1alpha (6-keto-PGF1alpha), alanine aminotransferase (ALT) activity, and the ultrastructure of liver tissue were dynamically observed.</p><p><b>RESULTS</b>Compared with the control group, SOD activity was significantly higher, LPO concentration, TXB2/6-keto-PGF1alpha ratio and ALT value were significantly lower (P < 0.05 and P < 0.01) in the LGT group, the PRO group and the LGT + PRO group during HIRI, with the abnormal changes of hepatic ultrastructure 25 min after reperfusion significantly alleviated in the three treated group.</p><p><b>CONCLUSION</b>Combination of ligustrazine and propofol shows protective effect on liver by decreasing oxygen free radical level, reducing lipid peroxidation and adjusting TXA2/PGI2 imbalance after hepatic ischemia-reperfusion in patients undergoing hepatic cancer surgery.</p>

Effect of propofol on anti-oxidation capability in the ischemia/reperfusion injury after hepatic ischemia in rabbits / 中国应用生理学杂志

<p><b>AIM</b>To investigate the role of oxygen free radicals (OFR) in hepatic ischemia and reperfusion injury (HI/RI) and effect of propofol on them.</p><p><b>METHODS</b>The rabbits were randomly divided into three groups (n=10), sham operated group (Control), HIR group(I/R) and HIR + propofol group (PRO). Changes of several parameters which included malondialdehyde (MDA), superoxide dismutase (SOD), xanthine oxidase (XO) and alanine aminotransferase(ALT) were measured before ischemia, 45 minutes after ischemia and 45 minutes after reperfusion in plasma. Meanwhile MDA concentration, SOD, XO activities and ALT value in liver tissue were measured, and the ultrastructure changes in liver tissue were observed under electron microscope at 45 minutes after reperfusion.</p><p><b>RESULTS</b>As compared with group control, XO, MDA and ALT increased and SOD decreased during HI/RI (P < 0.05 and P < 0.01) in plasma, and XO, MDA increased as well as SOD decreased at 45 minutes after reperfusion (P < 0.05 and P < 0.01) in liver tissue and there were abnormal changes of the hepatic ultrastructure at 45 minutes after reperfusion. Propofol reversed the results of mentioned indices as above markedly (P < 0.05 and P < 0.01).</p><p><b>CONCLUSION</b>OFR is an important factor during HI/RI, propofol may attenuate hepatic ischemia-reperfusion injury by dropping OFR level (raising SOD activity and dropping XO activity) and antagonizing lipid pe-reoxidation (reducing MDA content).</p>

Effects of ligustrazin on lipid peroxidation during hepatic ischemia reperfusion injury / 中国应用生理学杂志

<p><b>AIM</b>To explore the role of ligustrazin on dynamic changes of lipid peroxidation in hepatic ischemia/reperfusion injury (HIRI) and its mechanism.</p><p><b>METHODS</b>The HIRI model was used. Twenty rabbits were randomly divided into control group (n = 10) and ligustrazin group (n = 10). The xanthine oxidase (XO) activity, superoxide dismutase (SOD) activity,malondialdehyde (MDA) content and glutamic pyruvic transaminase (GPT) activity in plasma were observed before ischemia and at ischemia 25 min, reperfusion 25 min, reperfusion 60 min and reperfusion 120 min.</p><p><b>RESULTS</b>The XO activity, SOD activity, MDA content and GPT activity of ligustrazin group, as compared with control group, showed significant differences (P < 0.05 or P < 0.01) at total time points of reperfusion.</p><p><b>CONCLUSION</b>Ligustrazin has notable anti-lipid peroxidation effect on HIRI, which is due to its inhibiting the generation of oxygen free radicals and its strengthening scavenging of oxygen free radicals.</p>