Acute Toxicity and Hepatotoxicity of Aqueous Extracts of Taxilli Herba from Different Hosts in Zebrafish Model / 中国实验方剂学杂志

Objective:To explore the acute toxicities and hepatotoxicities of aqueous extracts of Taxilli Herba from <italic>Morus alba</italic>， <italic>Toxicodendron</italic> <italic>trichocarpum</italic>， <italic>Camellia oleifera</italic>， <italic>Salix babylonica</italic>， <italic>Melia azedarach</italic>， and <italic>Nerium indicum</italic> against zebrafish model and the effect of different hosts on the toxicity of Taxilli Herba， hoping to provide a theoretical basis for the safe use of Taxilli Herba. Method:The normally developed AB zebrafish at 3-day post fertilization was selected for acute toxicity study. According to the results of preliminary toxicity experiments， the zebrafishes were treated with aqueous extracts of Taxilli Herba from different hosts at six doses， and their mortality was calculated 72 h later. GraphPad Prism 6.0 was used for plotting the dose-toxicity curve， followed by the calculation of their median lethal concentration （LC₅₀） and 10% lethal concentration （LC₁₀）. The gz15Tg/+（AB） liver fluorescent protein transgenic zebrafish with normal development at 4-day post fertilization was applied for the hepatotoxicity study. The zebrafishes were divided into the low-， medium-， and high-dose groups of aqueous extracts of Taxilli Herba from six hosts， the positive control （acetaminophen） group， and the blank （embryo amniotic fluid） group， and then treated with the corresponding drugs. Seventy-two hours later， the liver morphology and fluorescent area changes in zebrafish were observed. And the activities of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） were detected. Result:The results of acute toxicity test demonstrated that the LC₅₀ values of water extracts of Taxilli Herba from <italic>M. alba</italic>， <italic>T.</italic> <italic>trichocarpum</italic>， <italic>C. oleifera</italic>， <italic>S. babylonica</italic>， <italic>M. azedarach</italic>， and <italic>N. indicum</italic> were 1.24， 0.94， 0.51， 0.38， 0.11， 0.09 g·L^-1， respectively， and the LC₁₀ values were 0.70， 0.60， 0.35， 0.28， 0.08， 0.07 g·L^-1， respectively. As revealed by hepatotoxicity test， compared with the blank group， the positive control group exhibited liver morphological changes， decreased fluorescent area （<italic>P</italic><0.01）， and elevated ALT and AST activities （<italic>P</italic>< 0.01）， suggesting that acetaminophen was hepatotoxic to zebrafish. However， there was no change in the liver morphology or fluorescent area of zebrafish in the low-， medium-， and high-dose groups of water extracts of Taxilli Herba from <italic>M. alba</italic>， and the ALT and AST activities were decreased. By contrast， the liver morphology and fluorescent areas in the medium- and high-dose groups of water extracts of Taxilli Herba from <italic>T.</italic> <italic>trichocarpum</italic>， <italic>C. oleifera</italic>， <italic>S. babylonica</italic>， <italic>M. azedarach</italic>， and <italic>N. indicum</italic> changed to varying degrees （<italic>P</italic><0.05， <italic>P</italic><0.01）. Besides， the activities of both ALT and AST were also enhanced. These indicated that Taxilli Herba from <italic>M. alba</italic> had no hepatotoxicity to zebrafish， while that from <italic>T.</italic> <italic>trichocarpum</italic>， <italic>C. oleifera</italic>， <italic>S. babylonica</italic>， <italic>M. azedarach</italic>， and <italic>N. indicum</italic> showed varying degrees of hepatotoxicity to zebrafish. Conclusion:The toxicity of Taxilli Herba is host-dependent. Taxilli Herba from <italic>M. alba</italic> has no hepatotoxicity， but that from the other five hosts shows varying degrees of hepatotoxicity. Standardizing the host source may be an important measure to realize the medication safety of Taxilli Herba.

Pharmacological Effect and Substance Basis of Neutral Blood-activating and Stasis-removing Traditional Chinese Medicine / 中国实验方剂学杂志

By reviewing the 2015 edition of Chinese Pharmacopoeia, the author collected 37 neutral blood-activating and stasis-removing traditional Chinese medicines. And by retrieving literatures on relevant material basis and pharmacological effects on CNKI, the author organized and summarized their chemical composition and pharmacological effects. Neutral blood-activating and stasis-removing traditional Chinese medicines are rich in chemical components, such as flavonoids, steroids and sugars. At the same time, it has a variety of pharmacological effects, including anti-platelet aggregation and anti-thrombotic mechanism, anti-atherosclerosis, inhibition of ischemia-perfusion injury, anti-tumor, anti-fibrosis, liver protection, anti-inflammatory analgesia, blood pressure reduction and immune regulation. It is widely used in the treatment of liver fibrosis, cardiovascular disease, liver cancer, uterine cancer, hypertension, hyperlipidemia and other diseases. Nowadays, as people has paid increasing attention to neutral herbs, studies on traditional Chinese medicines for activating blood circulation and removing blood stasis have been further deepened, which provides a better development prospect for neutral blood-activating and stasis-removing traditional Chinese medicines. From the perspective of medicinal properties, the authors systematically collected and summarized the pharmacological effects and material basis of neutral blood-activating and stasis-removing traditional Chinese medicines. This article provides theoretical guidance for the clinical application of neutral blood-activating and stasis-removing traditional Chinese medicines and new medicine research ideas for neutral blood-activating and stasis-removing traditional Chinese medicines.

Clinical Observation on Compound Huanggen Granules Combined with Entecavir Tablets in Treating Chronic Hepatitis B with Syndrome of Collateral Retardation Due to Blood Stasis / 中国实验方剂学杂志

Objective:To observe the clinical efficacy of compound Huanggen granules combined with Entecavir tablets for patients with chronic hepatitis B with syndrome of collateral retardation due to blood stasis. Method:The 130 patients with chronic hepatitis B with syndrome of collateral retardation due to blood stasis were randomly divided into observation group and control group by using random number table method. The 65 patients in observation group were treated with compound Huanggen granules combined with Entecavir tablets， and 65 patients in control group were treated with Entecavir tablets orally. The changes of liver function ［alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， total bilirubin（TBIL）， albumin（Alb）］ ， coagulation function ［prothrombin time（PT）， activated partial thromboplastin time（APTT）， thrombin time（TT）， fibrinogen（FIB）］， serum Hepatitis B Virus DNA（HBV DNA） levels， hepatitis B e antigen quantification（HBeAg）， stiffness of the liver， liver imaging （portal vein width， spleen thickness）， liver histopathology knodell HAI classification for chronic hepatitis， and changes in ishak fibrosis score at 24，48 weeks were observed in both groups. Result:After 48 weeks， the indexes of the two groups were improved to different degrees （P<0.05，P<0.01）， serum HBV DNA negative conversion rate， liver function， coagulation index， liver stiffness， portal vein diameter， Knodell HAI grade portal inflammation， interface activity， hepatic lobular activity， and Ishak fibrosis score in observation group were better than those in the control group （P<0.05）. Patients in the control group had no significant improvement in liver lobular activity after treatment in the Knodell HAI classification of chronic hepatitis. Conclusion:Compound Huanggen granules combined with Entecavir tablets are more effective in treating patients with chronic hepatitis B with syndrome of collateral retardation due to blood stasis than Entecavir tablets alone. The combination of two drugs helps to improve the rapid response rate of HBV DNA， and has a better effect on improving liver function， controlling the development of liver fibrosis and preventing related complications.

Correlation study on chemical constitutes of cardiac glycosides in Taxillus chinensis and its Nerium indicum host by UPLC-Q-TOF-MS/MS / 中国中药杂志

To build up an identification method on cardiac glycosides in Taxillus chinensis and its Nerium indicum host, and evaluate the influence on medicine quality from host to T. chinensis, ultra-performance liquid chromatography coupled with quadrupole-time-of-flight tandem mass-mass spectrometry(UPLC-Q-TOF-MS/MS)was applied. The samples of T. chinensis(harvested from N. indicum)and its N. indicum host were collected in field. The samples of T. chinensis(harvested from Morus alba)and its M. alba host was taken as control substance. All samples were extracted by ultrasonic extraction in 70% ethanol. Chromatographic separation was performed on an ACQUITY UPLC HSS T3 C_(18)(2.1 mm×100 mm,1.8 μm)column at 40 ℃. Gradient elution was applied, and the mobile phase was consisted of 0.1% formic acid water and acetonitrile. The 0.5 μL of sample solution was injected and the flow rate of the mobile phase was kept at 0.6 mL·min~(-1) in each run. It was done to identify cardiac glycosides and explore the chemical composition correlation in T. chinensis and its N. indicum host by analyzing positive and negative ion mode mass spectrometry data, elemental composition, cardiac glycoside reference substance and searching related literatures. A total of 29 cardiac glycosides were identified, 28 of it belonged to N. indicum host, 5 belonged to T. chinensis(harvested from N. indicum host), none of cardiac glycoside was identified in T. chinensis(harvested from M. alba host). The result could provide a reference in evaluating the influence in T. chinensis medicine quality from host. It was rapid, accurate, and comprehensive to identify cardiac glycosides in T. chinensis and its N. indicum host by UPLC-Q-TOF-MS/MS.