Biomechanopharmacology of Chinese Medicine Based on Mechano-ion Channel Piezo1： A Review / 中国实验方剂学杂志

Ischemic stroke is one of the leading causes of death and disability worldwide. In Han dynasty， HUA Tuo proposed the original preventive medicine idea that "with good blood circulation， the disease cannot be born"， which opened a broad space for the cross-research of blood-related mechanical factors and pharmacology. In the pathogenesis of ischemic stroke， mechanical factors comprehensively affect the function and crosstalk of platelets and endothelial cells. In recent years， as the well-known effects on thrombosis and stroke， more attention has been paid to hemodynamic factors as the participants involved in pathological mechanisms and potential therapeutic targets of ischemic stroke. The mechanical force ion channel Piezo1 widely exists on the surface of many types of cells. Besides being regulated by chemical and endogenous substances， Piezo1 responds to different mechanical conditions， regulates the opening and closing of channels， and activates different downstream signaling pathways. Piezo1 is now regarded as an important connection between mechanical and biochemical signals. A variety of Chinese medicine can affect the activity of Piezo1 protein， which may prevent and treat thrombotic diseases such as ischemic stroke through Piezo1 protein. In this paper， the effects of Piezo1 protein on the physiological and pathological functions of endothelial cells and platelet under different mechanical conditions and the role of Piezo1 in the process of thrombosis were reviewed， as well as the effects of Chinese medicine， chemical medicine， and endogenous substances targeting Piezo1 channel. These could provide new ideas for further exploring the mechanisms of Chinese medicines in activating blood circulation， developing new drugs， and deepening biomechanical-pharmacology research.

Establishment of Anti-inflammatory Activity Evaluation Method of Qingjin Huatantang Based on Phosphodiesterase Inhibitory Activity / 中国实验方剂学杂志

ObjectiveBased on the inhibitory activity of phosphodiesterase （PDE）， a method for determining the anti-inflammatory activity of Qingjin Huatantang was established to supplement and improve the quality control system of this famous classical formula. MethodHigh performance liquid chromatography （HPLC） was used to determine the activity of PDE， and the dose-effect relationship of inhibiting PDE activity of Qingjin Huatantang was investigated. The mobile phase consisted of methanol-0.5% acetic acid aqueous solution （5∶95）， and the detection wavelength was 254 nm. By measuring the PDE inhibition rate of multiple batches of Qingjin Huatantang water extract lyophilized powder， biological activity was marked with the activity of the neutralizing enzyme in the international unit U. ResultWhen the concentration of reaction substrate （cyclic adenosine monophosphate） was 50 μmol·L-1 and the reaction time was 60 min， the enzymatic reaction was stable with 4 U·mL-1 of PDE. In this reaction system， when the concentration of Qingjin Huatantang water extract lyophilized powder was 0.11-3.0 g·L-1， the inhibitory effect of PDE showed a concentration-dependent relationship. It was determined that the concentration of Qingjin Huatantang water extract lyophilized powder to be tested was 1 g·L-1， which showed a significant and stable inhibitory effect on PDE， and the inhibitory rate was >45%， that is， 1 mg of Qingjin Huatantang water extract lyophilized powder could neutralize the activity of 1.8 U PDE at least. ConclusionThis study establishes a biological activity evaluation method of Qingjin Huatantang based on the inhibitory activity of PDE， and the anti-inflammatory activity of Qingjin Huatantang is characterized by international unit U of PDE activity， which can provide a new method for the determination of biological activity of traditional Chinese medicine compounds.

Immunomodulatory and Antioxidant Activity of Gouqi Chewable Tablets / 中国实验方剂学杂志

ObjectiveTo explore the regulatory effect of Gouqi chewable tablets on innate and adaptive immunity in normal mice and its antioxidant activity in vitro and in vivo. MethodThe effects of low-， medium-， and high-dose groups （0.25， 0.5， 1.5 g·kg-1） on the immune function of normal mice were observed by carbon clearance test， immune organ index test， serum hemolysin test， ConA-induced splenic lymphocyte proliferation test， and natural killer cell （NK cell） activity test. The effects of Gouqi chewable tablets on the antioxidant capacity in vivo were determined by detecting the content of superoxide dismutase （SOD）， glutathione peroxidase （GSH-Px）， and malondialdehyde （MDA） in mice serum. The in vitro antioxidant activity of Gouqi chewable tablets was detected by 2，2'-azino-bis（3-ethylbenzothiazoline-6-sulfonic acid） diammonium salt （ABTS）， 2，2-diphenyl-1-picrylhydrazyl （DPPH）， and hydroxyl radical scavenging tests. ResultCompared with the blank control group， the low-， medium-， and high-dose groups of Gouqi chewable tablets improved the viability of NK cells， the proliferation of splenic lymphocytes， and the level of serum hemolysin antibody in mice （P<0.05）. The high-dose group increased the thymus index， spleen index， and phagocytic function of macrophages （P<0.05， P<0.01）. As compared with the blank control group， the activity of GSH-Px in mice serum in the medium-dose group was increased （P<0.05）， and the content of MDA in mice serum in the high-dose group was decreased （P<0.05）. In in vitro antioxidant tests， the median inhibitory concentration （IC50） values of Gouqi chewable tablets were 1.64±0.20， 2.04±0.03， and 10.27±0.03 g·L-1 by the DPPH， ABTS， and OH- free radical method， respectively. Those results indicated that Gouqi chewable tablets have good antioxidant effects in vitro. ConclusionGouqi chewable tablets can enhance the immune function of mice with good antioxidant effects.

Platelet function tests and Chinese medicines with antiplatelet function:a review / 中国中药杂志

Platelet function tests have been increasingly used to assist in the diagnosis of platelet disorders and prethrombotic state, monitoring of the efficacy of antiplatelet therapies, and personalized treatment. On the basis of light transmission aggregometry, new methods for platelet function test have been developed successively. At present, the research and development of platelet function detector is in its infancy in China. The active constituents of antiplatelet Chinese medicines can be classified into terpenoids, flavonoids, saponins, organic acids, lignans, diketones, volatile oils, and stilbenes. The results of dose-antiplatelet effect relationship of Chinese medicines and the active constituents showed that the effective concentration of the extracts or monomers of Chinese medicines was at micromolar level(μmol·L~(-1)), among which salvianolic acid B and ginkgolide K, ginkgolide B, and ginkgolide A had the strongest antiplatelet effect. These results suggest that the antiplatelet effect of Chinese medicine may be weaker than that of chemical drugs and biological products. Therefore, it is necessary to explore the structure-activity relationship of the active constituents in existing Chinese medicines and further improve their efficacy through structure modification. The antiplatelet effect of Chinese medicines and the constituents involves multiple pathways and multiple targets. These research results provide a reference for clinical application of them. However, there is still a lack of large-scale multi-center clinical trials to confirm the efficacy and safety of them. The regularity of the relationship between the structures of various constituents and their corresponding functions is still unknown and the relevant signal transduction pathways and structure-activity relationship need to be further studied. This paper summarized and analyzed the determination methods of platelet functions and the research results of antiplatelet Chinese medicines, which is of reference value for the research of effective and safe antiplatelet Chinese medicines.

Advance in Study on Classic Prescription Qingjin Huatantang / 中国实验方剂学杂志

Qingjin Huatanpang， first contained in Yixue Tongzhi， was composed of eleven medicinal materials of Scutellariae Radix， Gardeniae Fructus， Fritillariae Thunbergii Bulbus， Mori Cortex， Trichosanthis Semen Tostum， Citri Exocarpium Rubrum， Platycodonis Radix， Ophiopogonis Radix （core removed）， Anemarrhenae Rhizoma， Poria and Glycyrrhizae Radix et Rhizoma. It is a classic prescription created by YE Wen-ling in Ming dynasty for treating pulmonary disease with phlegm-heat obstructing lung syndrome. With the significant functions of clearing heat and moistening lung， reducing phlegm and relieving cough， it has been included in the "Classic Catalogue of Ancient Classics （First Batch）". Modern pharmacological studies have shown that Qingjin Huatantang has multiple activities such as relieving cough and eliminating phlegm， anti-inflammatory， bronchodilation， and immunoregulatory， and now it is commonly used for treating infectious lung diseases， such as acute exacerbation of chronic obstructive pulmonary disease， community acquire pneumonia， bronchiectasis， acute and chronic bronchitis in a form of its modified prescription or its combined use with western medicine， consistent with the clinical application in ancient times. According to the literatures on the study of Qingjin Huatantang published in recent years， this paper summed up the historical evolution， compatibility analysis， chemistry constituents， quality control， advances in pharmacology research， and clinical uses， which can provide theoretical and experimental data reference for further research and development， and proposed to establish a biological activity assay for quality control based on the pharmacological effect such as immunoregulatory activity， which can improve its quality control method and provide a reference for other famous classical formulas.

Suanzaoren Tang Treats Insomnia by Affecting Brain， Heart， and Liver Functions： A Review / 中国实验方剂学杂志

Sleep has been widely concerned by the medical field all over the world. Sleep deprivation can cause damage to organs of the human body， which is related to the occurrence of a variety of diseases. Besides， the pathological change in different organs of the human body is also a key factor that causes or aggravates insomnia. When treating insomnia and its complications， traditional Chinese medicine （TCM） focuses on the homology of the brain and heart， and insomnia is mainly treated from the five internal organs， especially the heart and liver. Sleep duration and structure change with age. The prevalence of insomnia is higher in older individuals susceptible to complications than in the younger population. In TCM， insomnia of blood deficiency and Yin deficiency is common among the elderly. Suanzaoren Tang is a classic prescription for nourishing blood and calming the mind and it is critical in the treatment of "sleeplessness due to consumptive disease and dysphoria"， with the effects of nourishing liver blood to calm the mind and clearing internal heat to relieve dysphoria. It has good efficacy on the insomnia of the elderly caused by deficiency of Qi and blood and abnormal operation of nutrient Qi and defense Qi. Furthermore， it also shows a certain therapeutic effect on insomnia combined with cardiovascular and cerebrovascular diseases. The present study revealed the damage to the brain， heart， and liver caused by sleep deprivation and the effect of Suanzaoren Tang on the brain， heart， and liver， and clarified the facts that Suanzaoren Tang inhibited the damage to organs caused by sleep deprivation and regulated energy metabolism， thereby exploring the sedative and hypnotic mechanism of Suanzaoren Tang to provide new ideas for Suanzaoren Tang in the treatment of sleep disorders and other diseases.

Effect of Suanzaoren Tang on Energy Metabolism of Liver Mitochondria in Aged Rats with Chronic Rapid Eye Movement Sleep Deprivation and Its Mechanism / 中国实验方剂学杂志

Objective:To study the effect of Suanzaoren Tang on energy metabolism of liver mitochondria in aged rats with chronic rapid eye movement （REM） sleep deprivation. Method:Fifty male Wistar rats were randomly divided into the control group， model group， estazolam group （0.18 mg·kg^-1·d^-1）， and low- （6.48 g·kg^-1·d^-1） and high-dose （12.96 g·kg^-1·d^-1） Suanzaoren Tang groups. Rats in all groups except for the control group received subcutaneous injection of <italic>D</italic>-galactose and then were deprived of sleep using the multiple platform method after the last administration. Following successful modeling， the rats in each group were treated with intragastric administration of the corresponding drugs for seven consecutive days. The morphology of liver mitochondria was observed under the transmission electron microscope. The content of adenosine triphosphate （ATP） in rat liver was detected by colorimetry， and the activities of ubiquinone oxidoreductase （complex Ⅰ）， succinate-ubiquinone oxidoreductase （complex Ⅱ）， ubiquinol-cytochrome c oxidoreductase （complex Ⅲ）， and cytochrome c oxidase （complex Ⅳ） in mitochondrial respiratory chain of rat liver were measured by colorimetry. The protein expression levels of citrate synthase （CS）， isocitrate dehydrogenase （IDH）， and ATP synthase， H⁺ transporting， mitochondrial F0 complex， subunit b， isoform 1 （ATP5F1） in rat liver were assayed by Western blot. Result:The mitochondrial damage in rat liver of the model group was more serious than that in the control group， manifested as mitochondrial swelling and deformation as well as cristae rupture and reduction. The comparison with the model group revealed that both the positive control and Suanzaoren Tang at the high dose obviously alleviated the mitochondrial swelling and deformation and reduced cristae rupture， with better improvements observed in the high-dose Suanzaoren Tang group. Compared with the control group， the content of ATP， the activities of mitochondrial respiratory chain complexes Ⅰ， Ⅱ， Ⅲ， and Ⅳ， and the protein expression levels of IDH， CS， and ATP5F1 in rat liver of the model group were all significantly decreased （<italic>P<</italic>0.01）. Compared with the model group， the content of ATP， the activities of mitochondrial respiratory chain complexes Ⅰ， Ⅱ， Ⅲ， and Ⅳ， and the protein expression levels of IDH， CS， and ATP5F1 in rat liver of the high-dose Suanzaoren Tang group were all significantly increased （<italic>P<</italic>0.05，<italic>P<</italic>0.01）. In the positive control group， the content of ATP， the activities of mitochondrial respiratory chain complexes I and Ⅲ， and the protein expression levels of CS and ATP5F1 in rat liver were significantly increased （<italic>P<</italic>0.05，<italic> P<</italic>0.01）. The activities of mitochondrial respiratory chain complexes Ⅰ and Ⅲ and the ATP5F1 protein expression in the low-dose Suanzaoren Tang group were significantly elevated （<italic>P<</italic>0.05， <italic>P<</italic>0.01）. Conclusion:Suanzaoren Tang alleviates the abnormal liver energy metabolism induced by chronic REM sleep deprivation in the elderly rats， which may be related to its enhancement of mitochondrial electron transport chain enzyme activities and the up-regulation of protein expression levels of CS， IDH and ATP5F1.

Effect of Suanzaoren Tang on Mitochondrial Damage and Apoptosis in Hypothalamus of Elderly Rats with Chronic Rapid Eye Movement Sleep Deprivation / 中国实验方剂学杂志

Objective:To investigate the effect of Suanzaoren Tang on mitochondria-mediated neuronal apoptosis. Method:Fifty male Wistar rats were randomly divided into the control group， model group， estazolam group （0.18 mg·kg^-1·d^-1）， and low- （6.48 g·kg^-1·d^-1） and high-dose （12.96 g·kg^-1·d^-1） Suanzaoren Tang groups. Rats in all groups except for the control group received subcutaneous injection of <italic>D</italic>-galactose and then were deprived of sleep using the multiple platform method after the last administration. Following successful modeling， the rats in each group were treated with intragastric administration of the corresponding drugs for seven consecutive days. The morphology of mitochondria in hypothalamus was observed under a transmission electron microscope. The activities of Na⁺-K⁺-ATPase and Ca²⁺-Mg²⁺-ATPase in hypothalamus were detected by spectrophotometry. Western blotting was conducted to determine the protein expression levels of cytochrome C （Cyt C）， B cell lymphoma-2 （Bcl-2）， Bcl-2 associated X protein （Bax） and cysteinyl aspartate-specific protease-3 （Caspase-3） in hypothalamus. Result:In the control group， there was no obvious pathological change in mitochondria， which were moderate in size and oval or spindle in shape， with the cristae arranged orderly. Compared with the control group， the model group exhibited abnormal mitochondrial morphology， manifested as obvious swelling， vacuolation， myelin figures， and cristae rupture and reduction. The comparison with the model group revealed that both the estazolam group and high-dose Suanzaoren Tang group alleviated the mitochondrial damage and reduced the vacuolation and swelling. Only some cristae rupture was present. The improvements were more obvious in the high-dose Suanzaoren Tang group. Compared with the control group， the activities of Na⁺-K⁺-ATPase and Ca²⁺-Mg²⁺-ATPase and the Bcl-2 protein expression in the model group were significantly decreased （<italic>P<</italic>0.01）， whereas the protein expression levels of Cyt C， Bax， and Caspase-3 were significantly increased （<italic>P</italic><0.01）. Compared with the model group， the activities of Na⁺-K⁺-ATPase and Ca²⁺-Mg²⁺-ATPase and the Bcl-2 protein expression in the estazolam group and the high-dose Suanzaoren Tang group were significantly elevated （<italic>P<</italic>0.01）， while the protein expression levels of Cyt C， Bax， and Caspase-3 were significantly lowered （<italic>P<</italic>0.05， <italic>P<</italic>0.01）. The activity of Na⁺-K⁺-ATPase and the Bcl-2 protein expression in the low-dose Suanzaoren Tang group were increased significantly （<italic>P<</italic>0.01）， but the Bax protein expression was down-regulated （<italic>P<</italic>0.05）. Conclusion:Suanzaoren Tang is able to improve the mitochondrial function of hypothalamic nerve cells and inhibit their apoptosis.

Effect of Suanzaoren Tang on Energy Metabolism of Myocardial Mitochondria in Aged Rats with Chronic Rapid Eye Movement Sleep Deprivation / 中国实验方剂学杂志

Objective:To study the mechanism of Suanzaoren Tang in regulating the energy metabolism of myocardial mitochondria in aged rats with chronic rapid eye movement （REM） sleep deprivation through the sirtuin 3 （SIRT3）/superoxide dismutase2 （SOD2） signaling pathway. Method:Fifty male Wistar rats were randomly divided into the control group， model group， estazolam group （0.18 mg·kg^-1·d^-1）， and low- （6.48 g·kg^-1·d^-1） and high-dose （12.96 g·kg^-1·d^-1） Suanzaoren Tang groups. Rats in all groups except for the control group received subcutaneous injection of <italic>D</italic>-galactose and then were deprived of sleep using the multiple platform method after the last administration. Following successful modeling， the rats in each group were treated with intragastric administration of the corresponding drugs for seven consecutive days. The morphology of myocardial mitochondria was observed under a transmission electron microscope. The content of adenosine triphosphate （ATP） in rat hypothalamus was detected by colorimetry， while the malondialdehyde （MDA） content and the SOD activity in myocardium were measured by spectrophotometry. Real-time polymerase chain reaction （Real-time PCR） and Western blot were conducted to determine the mRNA and protein expression levels of SIRT3 and SOD2 in rat myocardium. The localization of SIRT3 was detected by immunofluorescence staining. Result:Compared with the control group， the model group exhibited a disordered arrangement of myocardial filaments， accompanied by filament rupture and dissolution， obviously swollen mitochondria arranged in disorder， and blurring and even rupture of most mitochondrial cristae. Besides， the content of ATP and SOD activity in the myocardium decreased significantly （<italic>P<</italic>0.01）， whereas that of MDA increased significantly （<italic>P<</italic>0.01）. The mRNA and protein expression levels of SIRT3 and SOD2 were down-regulated significantly （<italic>P<</italic>0.01）， and the average fluorescence intensity of SIRT3 protein declined significantly （<italic>P<</italic>0.01）. The comparison with the model group revealed that high-dose Suanzaoren Tang enabled the myocardial filaments to be neatly arranged， relieved the mitochondrial damage and swelling， only manifested as partial mitochondrial cristae rupture， significantly increased ATP content， SOD activity， as well as SIRT3 and SOD2 mRNA and protein expression levels （<italic>P<</italic>0.01）， reduced the content of MDA （<italic>P<</italic>0.01）， and enhanced the average fluorescence intensity of SIRT3 protein （<italic>P<</italic>0.05）. The myocardial mitochondrial injury in the estazolam group was also alleviated. The activity of SOD and the SIRT3 and SOD2 mRNA and protein expression levels in the myocardium were significantly elevated （<italic>P<</italic>0.01）， while the activity of MDA was significantly lowered （<italic>P<</italic>0.01）. In the low-dose Suanzaoren Tang group， the improvement in myocardial mitochondrial injury was not obvious. However， both the SOD activity and SOD2 protein expression were significantly increased （<italic>P<</italic>0.05）. Conclusion:Suanzaoren Tang ameliorates the myocardial mitochondria injury and abnormal energy metabolism induced by chronic REM sleep deprivation in aged rats possibly by up-regulating the SIRT3 and SOD2 expression.

Biological Evaluation for Quality Control of Water Extract of Qingjin Huatantang Based on Phagocytic and Secretory Functions of Macrophages / 中国实验方剂学杂志

Objective:To establish a method for evaluating the biological activity of water extract lyophilized powder of Qingjin Huatantang based on the phagocytic and secretory functions of macrophages， and to control the quality of this formula from the biological activity level. Method:The phagocytic and inflammation models of RAW264.7 macrophages were established， the inhibition rates of water extract lyophilized powder of Qingjin Huatantang on interleukin-6 （IL-6） secretion and phagocytic index of neutral red of RAW264.7 macrophages were chosen as indicators to investigate the biological activity of Qingjin Huatantang， and the biological limit was searched. Result:The optimal inoculation density of RAW264.7 macrophages was 3×10⁵ pcs/mL， and the concentration of lipopolysaccharide （LPS） was 1 mg·L^-1 after treatment for 24 h. When the concentration was 500 mg·L^-1， water extract lyophilized powder of Qingjin Huatantang had no toxicity and no obvious promotion effect on the proliferation of RAW264.7 macrophages， and at this concentration， the phagocytosis of RAW264.7 macrophages for neutral red was significantly promoted， the phagocytic index was >113%. In addition， the lyophilized powder had a significant and stable inhibitory effect on IL-6 secretion of RAW264.7 macrophages induced by LPS， the inhibitory rate was >45%. Conclusion:Combined with the anti-inflammatory and immunomodulatory effects of Qingjin Huatantang， this study establishes an <italic>in vitro </italic>biological limit method for evaluating the quality of water extract of Qingjin Huatantang based on the phagocytic and secretory functions of RAW264.7 macrophages， and 500 mg·L^-1 was confirmed as the limit concentration. Under the limit concentration， Qingjin Huatantang water extract can significantly promote the phagocytic index of macrophages or significantly inhibit the secretion of IL-6 of RAW264.7 macrophages induced by LPS， which can be judged as qualified.

Preparation of paclitaxel-loaded and folic acid-modified poly (lactic-co-glycolic acid) nano-micelles and in vitro anticancer effect on cervical cancer HeLa cells / 中国中药杂志

The paclitaxel-loaded and folic acid-modified poly(lactic-co-glycolic acid) nano-micelles(PTX@FA-PLGA-NMs) were prepared by the emulsion solvent evaporation method, and the parameters of paclitaxel-loaded nano-micelles were optimized with the particle size and PDI as evaluation indexes. The morphology of the nano-micelles was observed by transmission electron microscopy(TEM), and the stability, drug loading and encapsulation efficiency were systematically investigated. In vitro experiments were performed to study the cytotoxic effects of nano-micelles, apoptosis, and cellular uptake. Under the optimal parameters, the nano-micelles showed the particle size of(125.3±1.2) nm, the PDI of 0.086±0.026, the zeta potential of(-20.0±3.8) mV, the drug loading of 7.2%±0.75%, and the encapsulation efficiency of 50.7%±1.0%. The nano-micelles were in regular spherical shape as observed by TEM. The blank FA-PLGA-NMs exhibited almost no inhibitory effect on the proliferation and growth of tumor cells, while the drug-loaded nano-micelles and free PTX exhibited significant inhibitory effects. The IC_(50) of PTX@FA-PLGA-NMs and PTX was 0.56 μg·mL~(-1) and 0.66 μg·mL~(-1), respectively. The paclitaxel-loaded nano-micelles were potent in inhibiting cell migration as assessed by the scratch assay. PTX@FA-PLGA-NMs had good pro-apoptotic effect on cervical cancer HeLa cells and significantly promoted the uptake of HeLa cells. The results of in vitro experiments suggested that PTX@FA-PLGA-NMs could target and treat cervical cancer HeLa cells. Therefore, as nanodrug carriers, PTX@FA-PLGA-NMs with anti-cancer activity are a promising nano-system for improving the-rapeutic effects on tumors.

On shear-induced platelet aggregation and its therapy / 药学学报

It is now widely accepted that platelet aggregation plays an important role in physiological hemostasis and pathological thrombosis associated with cardiovascular and cerebrovascular diseases. Anti-platelet aggregation drug research is also a hot spot of current research. The biggest challenge of antiplatelet therapy has been the molecular overlap of the hemostasis and thrombosis, leading to a serious risk of bleeding. Recent studies have emphasized the importance of shear stress generated from blood flow, which will primarily drive platelet activation and aggregation in thrombosis. So if we can take advantage of the differences between the physiological and pathological vascular blood flow environment, the development of selective anti-platelet therapy may be a safer treatment for cardiovascular and cerebrovascular diseases. In this review, we discuss the underlying mechanisms of shear-induced platelet activation. Later, we summarize the effects and mechanisms of compounds and traditional Chinese medicine on shear-induced platelet activation. The aim is to provide a reference for the study of biological pharmacology of traditional Chinese medicine for promoting blood circulation and removing blood stasis.

Effects of Xueshuantong Injection on thrombosis formation and blood flow in rats / 中国中药杂志

The aim of this paper was to explore the effect of Xueshuantong Injection(freeze-dried powder,XST) on κ-carrageenan-induced thrombosis and blood flow from the aspects of interactions among blood flow,vascular endothelium and platelets. Fifty male Sprague-Dawley rats(190-200 g) were randomized into five groups: control group, model group, heparin sodium(1 000 U·kg~(-1)) group, low-dose and high-dose(50, 150 mg·kg~(-1)) XST groups. Rats were intraperitoneally injected with corresponding drugs and normal saline(normal control and model groups) for 10 days. One hour after drugs were administered intraperitoneally on the 7 th day, each rat was injected with κ-carrageenan(Type Ⅰ, 1 mg·kg~(-1)) which was dissolved in physiological saline by intravenous administration in the tail to establish tail thrombus model. The lengths of black tails of the rats were measured at 2, 6, 24 and 48 h after modeling. Vevo®2100 small animal ultrasound imaging system was used to detect the internal diameter of rat common carotid artery, blood flow velocity and heart rate, and then the blood flow and shear rate were calculated. Meanwhile, the microcirculatory blood flow perfusion in the thigh surface and tail of rats were detected by laser speckle blood flow imaging system. Platelet aggregometry was used to detect the max platelet aggregation rate in rats. Pathological changes in tail were observed through hematoxylin-eosin staining, and Western blot was used to detect the protein content of platelet piezo1. According to the results, XST could inhibit the rat tail arterial thrombosis and significantly reduce the length of black tail(P<0.05). The blood flow of common carotid artery in XST low dose group was significantly higher than that in the model group(P<0.05). XST high dose group could significantly increase the microcirculatory blood flow perfusion of the tail in rats as compared with the model group(P<0.05). XST high dose group could significantly inhibit platelet aggregation rate(P<0.05) and XST low dose group could significantly inhibit platelet piezo1 protein expression(P<0.01). In summary, XST could play an effect in fighting against thrombosis induced by κ-carrageenan in rats, which may be related to significantly inhibiting platelet aggregation, improving body's blood flow state, maintaining normal hemodynamic environment and affecting mechanical ion channel protein piezo1.

Feasibility of echocardiography-guided repeated intraventricular blood sampling in mice / 中华心血管病杂志

Objective: To investigate the feasibility of echocardiography-guided closed-chest repeated intraventricular blood sampling in mice, and to clarify the maximum blood volume that can be collected by this method, and whether the method can be used for long-term repeated blood collection in mice. Methods: Twenty-four male C57BL/6J mice (10-14 weeks old) were divided into the terminal experiment group (n=4, for investigating the maximum blood amount that could be sampled at one time), the repeated 0.5 ml blood collection group (n=10, sampling 0.5 ml whole blood each time, once every two days for consecutive 4 weeks), and the repeated 0.75 ml blood collection group (n=10, sampling 0.75 ml whole blood each time, once every two days for consecutive 4 weeks). High-frequency echocardiography was used to display the largest section of the left ventricle, guiding the insulin syringe needle through the thorax into the left ventricle for blood collection. In the repeated 0.5 ml blood collection group, echocardiography was used to detect the cardiac structure and function before blood collection, three minutes after blood collection, and one week after the last (the 14th) blood collection. Results: We successfully performed echocardiography-guided closed-chest intraventricular blood sampling, with an average operating time (88±19)s per mouse, and a maximum blood volume (1.43±0.11)ml per mouse. In the repeated 0.5 ml blood collection group, heart rate, left ventricular ejection fraction, left ventricular fractional shortening, left ventricular end-diastolic dimension and left ventricular posterior wall end-diastolic thickness remained uncganged before the first blood collection and after 4 weeks of repeated blood collection (all P>0.05). No death in the repeated 0.5 ml blood collection group. However, in the 0.75 ml blood collection group, two mice died before the end point. Conclusions: The echocardiography-guided closed-chest intraventricular blood sampling is a safe, minimally invasive, convenient and efficient method, and can be used repeatedly for long-term blood collection in mice.

Effect of Shenghuitang on Bmal1 in Hypothalamus and IL-6 and TNF-α in Hippocampus of APP/PS1 Double Transgenic Dementia Model Mice / 中国实验方剂学杂志

Objective:To investigate the effect of Shenghuitang on learning and memory, biological clock gene[brain and muscle arnt-like 1 (Bmal1)] in hypothalamus and interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in hippocampus of APP/PS1 double transgenic dementia model mice, in order to explore the possible mechanism of Shenghuitang to improve learning and memory and sleep disorders. Method:The experimental mice were randomly divided into model group, blank control group, melatonin group, high-dose Shenghuitang group and low-dose Shenghuitang group. Autonomic activity analysis system was used to detect the autonomic activities of mice in each group. Morris water maze was used to detect the learning ability and spatial memory ability of each group. quantitative real-time fluorescence polymerase chain reaction(Real-time PCR) was used to detect the expression of Bmal1 mRNA in the hypothalamic area of mice. Western blot was used to detect the expression of Bmal1 protein in each group. The content of inflammatory factors IL-6 and TNF-α in hippocampus of mice was detected by enzyme-linked immunosorbent assay(ELISA). The correlation between inflammatory factors IL-6, TNF-α and Bmal1 gene was analyzed by pearson analysis. Result:The results of voluntary activities showed that compared with the control group, the number of activities and activity distance of the model group were significantly decreased (PPPPPPPPPPPPPPα in the model group were significantly higher than those in the control group (Pα in the drug group were significantly lower(Pα and Bmal1 were correlated and negatively correlated. Conclusion:Shenghuitang may reduce the levels of inflammatory factors IL-6 and TNF-α in hippocampus by up-regulating the expression of Bmal1 gene in hypothalamic region, thus improving Alzheimer' s disease(AD) and circadian rhythm disorders.

Effect of Shenghuitang on Learning Memory, Levels of IL-6, TNF-α and COX-2 in Hippocampus of Chronic Sleep Deprivation Mice / 中国实验方剂学杂志

Objective: To observe the effect of Shenghuitang on learning and memory and expressions of interleukin-6(IL-6), tumor necrosis factor-α(TNF-α) and cyclooxygenase-2(COX-2) in hippocampus of chronic sleep deprived mice, in order to explore the possible mechanism of Shenghuitang in improving learning and memory ability. Method: Mice were randomly divided into sleep deprivation group, blank group, melatonin group(7.8×10-4 g·kg-1·d-1), high, middle and low-dose Shenghuitang groups(54,27,13.5 g·kg-1·d-1). The model of chronic sleep deprivation in mice was established using the "multi-platform water environment method". 28 d sleep deprivation and intragastric administration were provided. Morris water maze was used to detect the learning and memory ability of mice in each group. Real time-PCR was used to detect mRNA expressions of IL-6, TNF-α and COX-2 in the hippocampus of each group. Result: The results of Morris water maze test showed that compared with the blank group, the total time spent on finding the platform and the total swimming distance of the model group were significantly prolonged (PPPPPPPPPα, and COX-2 were increased in the model group compared with the blank group. Compared with the model group, mRNA expressions of IL-6, TNF-α, and COX-2 were decreased in the treated group. COX-2 mRNA expression was down-regulated. Conclusion: Shenghuitang may improve the learning and memory ability of mice by decreasing mRNA expressions of IL-6, TNF-α and COX-2 in hippocampus.

Development of bioassay method in quality control of traditional Chinese medicine / 中国中药杂志

On the basis of chemical content determination， the bioassay methods can be used to comprehensively evaluate and control the Chinese medicine compound. This paper analyzed the newly published literature on traditional Chinese medicine(TCM) bioassay. The selection of standard control substances and the establishment of experimental system are the main difficulties in bioassay. At present， the standard control substances mainly include： different sources of products with basically similar components， certified medicinal materials， genuine medicinal materials， commonly used chemical drugs or biological products with similar pharmacological functions， as well as Chinese medicine potency conversed by activity of biological products. In this paper， the common bioassays would be summarized from the clinical efficacy of activating blood circulation and removing stasis and clearing heat and detoxification. It is one of the important contents in the industrial production of traditional Chinese medicine to gradually establish the bioassay platform of Chinese medicine from the enterprise's internal control to the industry. recognition.

Network pharmacology deciphering multiple mechanisms of volatiles of Wendan granule for treatment of senile dementia / 中国药理学与毒理学杂志

OBJECTIVE To explore the mechanisms of the volatiles of Wendan granule for the treatment of senile dementia,network pharmacology method integrating absorption,distribution,metab-olism, and excretion (ADME) screening, target fishing, network constructing, pathway analyzing, and correlated diseases prediction was applied. METHODS Twelve small molecular compounds of WDG were selected as the objects from 74 volatiles with the relative abundances above 2%,and their ADME parameters were collected from Traditional Chinese Medicine Systems Pharmacology platform (TCMSP), and then the corresponding targets, genes, pathways and diseases were predicted according to the data provided by TCMSP,DrugBank,Uniport and the Database for Annotation,Visualization and Integrated Discovery(DAVID).The related pathways and correlation analysis were explored by the Kyoto Encyclo-pedia and Genomes (KEGG) database. Finally, the networks of compound-target, target-pathway and pathway-disease of WDG were constructed by Cytoscape software. RESULTS Twelve compounds interacted with 49 targets, of which top three targets were Gamma-aminobutyric acid receptor subunit alpha-1 (GABRA1), Prostaglandin G/H synthase 2 (PGHS-2) and Sodium-dependent noradrenaline transporter.Interestingly,these targets were highly associated with depression,insomnia and Alzheimer′s disease that mainly corresponded to mental and emotional illnesses. CONCLUSION The integrated network pharmacology method provides precise probe to illuminate the molecular mechanisms of volatiles of WDG for relieving senile dementia related syndromes,which will also facilitate the application of traditional Chinese medicine in modern medicine,as well as follow-up studies such as upgrading the quality stan-dard of clinical medicine and novel drug development.

An epidemiological survey of ocular fundus diseases in primary and secondary school students of Dagang Oilfield in Tianjin / 天津医药

Objective To study the fundus diseases that primary and middle school students may be susceptible to suffer, and to understand the prevalence rate and constituent ratio of these diseases. Methods A total of 9 504 students were examined from November 2015 to February 2017 to find out their ocular fundus conditions. They were from all the primary and secondary schools directly subordinate to Dagang Oilfield headquarters. Among the 9 504 participants 4 998 were male and 4 506 were female,with the age span of 5 to 23 and the average age of(13.40±3.41).Our medical examination included uncorrected visual acuity examination and fundus oculi photography. The distribution of fundus diseases was analyzed,including gender,vision acuity and age composition of patients with major diseases. Results Among the 9 504 students,4 314 had emmetropic eyes,and 5 190 had refractive errors.A total of 9 126 out of the 9 504 examined students had normal ocular fundus(96.02%),while those with abnormal ocular fundus accounted for 378(3.98%).The main types of abnormal ocular fundus included the tessellation fundus 354 (3.72%), followed by the large depression of optic disk 16 (0.17%). Among all the 354 cases with tessellation fundus, 179 were male and 175 were female. There were 257 students whose visual acuity≤0.3/0.3,and the prevalence was higher in 17-19 years old students(5.86%).Conclusion It has been revealed in this epidemiological survey that the main abnormal ocular fundus of primary and middle school students in Dagang is tessellation fundus.Most of the tessellation fundus are caused by middle and high myopia,and very few are found in emmetropic students. It has also been found that the incidence of tessellation fundus increases significantly with the increase of age. The large depression of optic disk ranks the second in the abnormal ocular fundus, which is mostly physiological.

Influence of Periampullary Diverticulum on Common Bile Duct Stone / 胃肠病学

Background:With the progress and widely application of endoscopic techniques,the prevalence rate of periampullary diverticulum (PAD)is increasing in recent years. However,the results of studies investigating the correlation of PAD and its types with common bile duct (CBD)stone are different. Aims:To investigate the influence of PAD and its types on CBD stone. Methods:A total of 1524 consecutive inpatients who underwent diagnostic and therapeutic ERCP for pancreatobiliary diseases from Jan. 2014 to Dec. 2016 at the General Hospital of Xinjiang Military Region of Chinese PLA were enrolled and divided into two groups according to the presence or absence of PAD. Patients in PAD group were further classified into 3 subgroups by the papilla's location with respect to the diverticulum. Their clinical data were collected and retrospectively analyzed. Results:The proportion of elderly patients (≥60 years old)in PAD group was 82. 2% (310 /377),which was significantly higher than that in non-PAD group [60. 8% (697 / 1147),P < 0. 05]. The prevalences of CBD stone,gallstone associated with CBD stone,post-cholecystectomy and recurrent CBD stone were higher and the size of CBD stone was larger in PAD group than in non-PAD group (P all < 0. 05). Furthermore,stratified analysis revealed that the CBD stone was more prevalent and the size of stone was larger in type Ⅱ PAD than in type Ⅰ and type Ⅲ PAD (P <0. 05), while recurrent CBD stone was more frequent in type Ⅰ and type Ⅱ PAD than in type Ⅲ PAD (P < 0. 05). Multivariate analysis indicated that the elderly and PAD were the risk factors for recurrence of CBD stone,and cholecystectomy was a protective factor. Conclusions:The elderly is predisposed to PAD. PAD and its types are associated with the development and recurrence of CBD stone. CBD stone is more prevalent in patients with PAD especially type Ⅱ PAD. The size of stone is larger and the post ERCP recurrence rate is high in type Ⅱ PAD.