Abstract：ObjectiveTo investigate the effect of Liuwei Dihuangwan on memory function of senescence-accelerated mouse prone 8 （SAMP8） mice by regulating autophagy through the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/forkhead box O3a （FoxO3a） pathway.MethodSix male senescence-accelerated mouse resistant 1 （SAMR1） mice of SPF grade aging 6 months were assigned to a normal group， and 24 male SAMP8 mice of SPF grade aging 6 months were randomly divided into a model group， a donepezil group （0.747 mg·kg-1）， and high- and low-dose Liuwei Dihuangwan groups （2.700 and 1.350 g·kg-1）， with 6 mice in each group. The mice were treated with drugs by gavage for 2 months. Morris water maze was used to detect the learning and memory abilities of mice in each group. Nissl staining was used to observe the neurons in the cortex and hippocampus. The positive expression of microtubule-associated protein 1 light chain 3B （LC3B） in the cortex and hippocampus was detected by immunohistochemistry （IHC）. Western blot was used to detect the protein expression of the mammalian ortholog of yeast ATG6 （Beclin-1）， B cell lymphoma-2 （Bcl-2）， autophagy-related gene 5 （ATG5）， cysteinyl aspartate-specific protease 3 （Caspase-3）， Caspase-9， Akt， p-Akt， FoxO3a， and p-FoxO3a.ResultCompared with the normal group， the model group showed prolonged escape latency （P<0.05，P<0.01）， reduced number of platform crossings and the residence time in the target quadrant （P<0.01）， decreased neurons with reduced volume and dispersed distribution in the cortex and hippocampus， increased positive expression of LC3B （P<0.01）， elevated expression of Beclin-1 and ATG5 in the cortex （P<0.01）， declined Bcl-2 expression （P<0.01）， up-regulated Caspase-3 and Caspase-9 expression （P<0.01）， and decreased expression levels of p-Akt/Akt and p-FoxO3a/FoxO3a （P<0.01）. Compared with the model group， the donepezil group and the Liuwei Dihuangwan groups showed shortened 3 d escape latency （P<0.05，P<0.01）， increased number of platform crossings （P<0.01）， and prolonged residence time in the target quadrant （P<0.01）. In the donepezil group， the number of neurons in the cortex and hippocampus was increased. In the Liuwei Dihuangwan groups， the number of neurons and Nissl bodies increased with denser distribution and lower degree of cell damage. The positive expression of LC3B in the cortex and hippocampus was decreased in the donepezil group and Liuwei Dihuangwan groups （P<0.01）. The expression of Beclin-1 was decreased in the Liuwei Dihuangwan groups （P<0.01）. The expression of ATG5 was decreased in the donepezil group and the low-dose Liuwei Dihuangwan group （P<0.01）. The donepezil group and the Liuwei Dihuangwan groups showed the increased expression level of Bcl-2 in the cortex （P<0.01）， decreased expression level of Caspase-3 （P<0.01）， reduced expression level of Caspase-9 （P<0.05，P<0.01）， and elevated expression levels of p-Akt/Akt and p-FoxO3a/FoxO3a （P<0.01）.ConclusionLiuwei Dihuangwan can effectively improve the learning and memory abilities of the SAMP8 mice and protect neurons. Its mechanism may be related to the regulation of the PI3K/Akt/FoxO3a signaling pathway， down-regulation of the expression of ATG5， Beclin-1， and LC3B， and the inhibition of apoptosis.
Abstract：ObjectiveTo study the effect of Buyang Huanwutang on Kelch-like Ech-related protein 1 （Keap1）/nuclear factor E2-related factor 2 （Nrf2）/heme oxygenase-1 （HO-1） antioxidant signaling pathway in rats with idiopathic pulmonary fibrosis （IPF） and explore the mechanism of this prescription in the treatment of IPF.MethodForty SPF-grade male SD rats were assigned into a sham operation group， a model group， a Buyang Huanwutang group， and a nintedanib group according to random number table method， with 10 rats in each group. IPF rat model was established by intratracheal infusion of bleomycin （0.005 g·kg-1） in other groups except the sham operation group. Buyang Huanwutang group was administrated with Buyang Huanwutang (14.84 g·kg-1)，intragastric administration of nitedanib suspension (0.1 g·kg-1)，sham operation group and model group were given equal volume of normal saline， for 28 days. After lung function test， serum and lung tissue samples were collected. Hematoxylin-eosin （HE） staining and Masson trichrome staining were employed to observe the pathological changes of the lung tissue. The content of hydroxyproline （HYP） in lung tissue was detected. The levels of malondialdehyde （MDA） in serum and lung tissue， and the activities of superoxide dismutase （SOD）， glutathione peroxidase （GSH-Px） and catalase （CAT） were determined. The mRNA and protein levels of Keap1， Nrf2， and HO-1 was determined by Real-time fluorescent quantitative polymerase chain reaction， immunohistochemical staining， and Western blot.ResultCompared with the sham operation group， the modeling increased the resistance and elasticity and decreased the compliance of respiratory system （P<0.01）， elevated the lung index， pathological score， and HYP content in lung tissue （P<0.01）， and enriched MDA in serum and lung tissue， while it decreased the activities of SOD， GSH-Px， and CAT （P<0.01）. Furthermore， the modeling down-regulated the mRNA and protein levels of Keap1 and up-regulated those of Nrf2 and HO-1 in lung tissue （P<0.01）. Compared with the model group， Buyang Huanwutang decreased the resistance and elasticity and increased the compliance of respiratory system （P<0.01）， lowered the lung index， pathological score， and HYP content in lung tissue （P<0.01）， and reduced MDA in serum and lung tissue， while it increased the activities of SOD， GSH-Px， and CAT （P<0.01）. Additionally， Buyang Huanwutang down-regulated the expression of Keap1 and up-regulated that of Nrf2 and HO-1 in lung tissue （P<0.05， P<0.01）.ConclusionBuyang Huanwutang can activate Keap1/Nrf2/HO-1 signaling pathway to enhance the antioxidant capacity and slow down the pathological process of IPF in rats.
Abstract：ObjectiveTo investigate the regulatory effect of Shoutaiwan on oxidative stress and pyroptosis in lipopolysaccharide （LPS）-induced human extravillous trophoblast （HTR-8/SVneo） cells and provide a new direction for deciphering the mechanism of action of Shoutaiwan.MethodLPS （100 μg∙L-1） was used to induce the injury of HTR-8/SVneo cells （modeling）. Five groups were designed in this study， including a blank group， a model group， a Shoutaiwan （10% Shoutaiwan-containing serum） group， an antioxidant （1 mmol·L-1 NAC） group， and NOD like receptor thermoprotein domain 3 （NLRP3） inhibitor （50 μmol·L-1 MCC950） group. Cell viability was detected by cell counting kit-8 （CCK-8） kit. Hochest 33342/PI double fluorescence staining and flow cytometry were employed to observe cell death. The levels of interleukin-18 （IL-18）， interleukin-1β （IL-1β）， malondialdehyde （MDA）， and superoxide dismutase （SOD） in cell supernatant was determined by enzyme-linked immunosorbent assay （ELISA）. DCFH-DA probe was used to measure the level of intracellular reactive oxygen species （ROS）. Western blot was employed to determine the protein levels of NLRP3， Caspase-1， gastermin D （GSDMD）， and IL-1β in cells， and Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） to measure the mRNA levels of NLRP3 and Caspase-1 in cells.ResultCompared with the blank group， the modeling decreased the cell viability （P<0.01）， elevated the levels of IL-1β， IL-18， ROS， and MDA， and weakened the activity of SOD （P<0.01）. Furthermore， it up-regulated the protein levels of NLRP3， Caspase-1， GSDMD， and IL-1β and the mRNA levels of NLRP3 and Caspase-1 （P<0.01）. Compared with the model group， Shoutaiwan， NAC， and MCC950 increased the cell viability （P<0.01）. Further， Shoutaiwan and NAC lowered the levels of MDA and ROS and increased the activity of SOD （P<0.01）. Shoutaiwan and MCC950 reduced the IL-1β and IL-18 in cell supernatant （P<0.01）， and down-regulated the protein levels of NLRP3， Caspase-1， GSDMD， and IL-1β and the mRNA levels of NLRP3， Caspase-1， and IL-1β （P<0.05，P<0.01）.ConclusionShoutaiwan can regulate oxidative stress and pyroptosis to attenuate the LPS-induced damage of HTR-8/SVneo cells， which may be the mechanism of Shoutaiwan in preventing recurrent spontaneous abortion.
Abstract：ObjectiveTo study the effect of Siwutang （SWT） on intestinal flora in rats with diminished ovarian reserve （DOR） induced by Tripterygium wilfordii polyglycoside （TWP） based on 16S rRNA sequencing.MethodTwenty 8-week-old female SD rats were randomly assigned into four groups： blank group， model group， SWT high-dose group， and SWT low-dose group. Except the blank group， the other three groups were underwent intragastric administration of TWP tablets at a dose of 50 mg·kg-1 for 14 days. On day 15， the high-dose group was administrated at 3 times of the human dosage （40 g/person/day）， the low-dose group at 1.5 times of the human dosage， and the model group and the blank group with the same volume of normal saline for 18 days. Then， feces were collected for 16S rRNA sequencing. One hour after administration， blood was sampled from abdominal aorta after anesthesia for the measurement of hormone levels by radioimmunoassay， and ovaries were sampled， embedded， sliced， and stained with haematoxylin-eosin （HE） for pathological observation.ResultThe model group had higher level of luteinizing hormone （LH， P<0.05） and lower level of estradiol （E2， P<0.05） than the blank group. The SWT high-dose group and low-dose group had lower LH levels （P<0.05） and higher E2 levels than the model group （P<0.05）. SWT reversed the elevation in follicle-stimulating hormone （FSH） and LH levels and the decline in E2 and progesterone （P） levels caused by TWP to some extent. There were a large number of follicles at different developmental stages in the blank group， while atretic follicles increased significantly in the model group. A large number of mature follicles， secondary follicles， and primary follicles were observed in the high-dose SWT group， and primordial follicles， secondary follicles， and increased corpus luteum in the low-dose SWT group. Compared with that in the blank group and the administration group， the abundance of Verrucomicrobia and Epsilonbacteraeota in the model group significantly reduced. Compared with the blank group， the model group had different intestinal flora in phylum， class， order， family， and genus levels. Specifically， the model group had increased proportions of Firmicutes and Bacteroidetes. After TWP modeling， the abundance of Lachnospiraceae decreased significantly while that of Ruminococcaceae UCG-005 increased significantly. SWT groups， blank group， and model group can be clearly distinguished， and SWT groups had a tendency to approach the blank group.ConclusionSWT may improve the ovarian function of rats with TWP-induced DOR by regulating intestinal flora diversity.
Abstract：ObjectiveTo explore the mechanism of Qihuang Yiqi Shexue prescription （QHYQSX） in the treatment of immune thrombocytopenia （ITP） model mice based on the autophagy mediated by the adenosine monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR）/Unc-51-like kinase 1 （ULK1） signaling pathway.MethodFifty BALB/c mice were randomly divided into normal group， model group， high- and low-dose QHYQSX groups， and prednisone group， with 10 mice in each group. The ITP model was induced by intraperitoneal injection of anti-platelet serum （APS） of guinea pig. On the 8th day of the APS injection， drugs were administered by gavage for 14 days. Peripheral blood platelet （PLT） count and hemoglobin （Hb） concentration were detected. Spleen and thymus were separated， weighed， and the organ index was calculated. Sternum was sampled for bone marrow smear， and bone marrow megakaryocytes were classified under a microscope. Thrombopoietin （TPO）， interleukin-6 （IL-6）， IL-10， tumor necrosis factor-α （TNF-α）， transforming growth factor-β1 （TGF-β1）， and interferon-γ （IFN-γ） in the serum were detected by enzyme-linked immunosorbent assay（ELISA）. AMPK， mTOR， ULK1， microtubule-associated protein light chain 3 （LC3）， Beclin1， and p62 mRNA expression levels in the spleen were detected by Real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR）. The protein expression of AMPK， p-AMPK， p-mTOR， p-ULK1， LC3Ⅱ/LC3Ⅰ， Beclin1， and p62 in the spleen was detected by Western blot.ResultCompared with the normal group， the model group showed reduced peripheral blood PLT count， Hb， and TPO levels （P<0.05，P<0.01）， increased spleen and thymus indexes （P<0.01）， decreased number of bone marrow megakaryocytes （P<0.01）， elevated serum levels of IL-6， TNF-α， and IFN-γ （P<0.01）， and reduced IL-10 and TGF-β1 levels （P<0.01）. Compared with the model group， the groups with drug intervention showed increased PLT counts and TPO levels （P<0.01）， decreased spleen and thymus indexes （P<0.05， P<0.01）， elevated number of bone marrow megakaryocytes （P<0.05， P<0.01）， reduced serum levels of IL-6， TNF-α， and IFN-γ （P<0.05， P<0.01）， and up-regulated IL-10 and TGF-β1 levels （P<0.05，P<0.01）. Compared with the low-dose QHYQSX group， the high-dose QHYQSX group and the prednisone group showed different degrees of significant differences in improving PLT counts and levels of cellular inflammatory factors （P<0.05， P<0.01）. Real-time PCR and Western blot results showed that compared with the normal group， the model group showed up-regulated mRNA expression of AMPK， LC3， and Beclin1 and protein expression of p-AMPK/AMPK， LC3Ⅱ/LC3Ⅰ， and Beclin1 in the spleen （P<0.05， P<0.01）， and down-regulated mRNA expression of mTOR， ULK1， and p62 and protein expression of p-mTOR， p-ULK1， and p62 （P<0.05， P<0.01）. Compared with the results in the model group， high- and low-dose QHYQSX and prednisone could down-regulate the mRNA expression of AMPK， LC3， and Beclin1 and protein expression of p-AMPK/AMPK， LC3Ⅱ/LC3Ⅰ， and Beclin1 in the spleen （P<0.05， P<0.01）， and up-regulate the mRNA expression of mTOR， ULK1， and p62 and protein expression of p-mTOR， p-ULK1， and p62 （P<0.05， P<0.01）.ConclusionQHYQSX may inhibit excessive autophagy by regulating the AMPK/mTOR/ULK1 signaling pathway， thereby regulating immune intolerance and playing a role in the treatment of ITP.
Keywords：Qihuang Yiqi Shexue prescription;immune thrombocytopenia （ITP）;autophagy;adenosine monophosphate-activated protein kinase/mammalian target of rapamycin/Unc-51-like kinase 1（AMPK/mTOR/ULK1） signaling pathway;inflammatory reaction
Abstract：ObjectiveTo reveal the effect of Wenxin prescription on mitochondrial energy metabolism and silent information regulator 1 （SIRT1）/peroxisome proliferator-activated receptor-γ coactivator-1α （PGC-1α）/recombinant estrogen-related receptor α （ERRα） signaling pathway in rats with myocardial ischemia-reperfusion injury.MethodTotally 90 male Wistar rats of SPF grade were randomly assigned into a sham operation group， a model group， and low-， medium-， and high-dose Wenxin prescription groups， with 18 rats in each group. The rats in low-， medium-， and high-dose Wenxin prescription groups were administrated with 0.99， 1.98， and 3.96 g·kg-1 granules by gavage， respectively， and those in the sham operation group and model group with the same amount of normal saline. Twenty-one days after pre-administration， the rat model of myocardial ischemia-reperfusion injury was established by ligation of the left anterior descending coronary artery for 30 min and reperfusion for 2 h， and the rats in the sham operation group were only threaded without ligation. Myocardial infarction area was observed through 2，3，5-triphenyl-2h-tetrazolium chloride （TTC） staining， and the myocardial histopathology through hematoxylin-eosin （HE） staining. The levels of creatine kinase-MB （CK-MB） and lactate dehydrogenase （LDH） in serum， cytochrome C oxidase （CCO） and succinate dehydrogenase （SDH） in mitochondrion， and ATP in myocardial tissue were detected according to kit instructions. The mRNA and protein levels of SIRT1， PGC-1α， ERRα， and mitochondrial transcription factor A （TFAM） in myocardial tissue were determined by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively.ResultCompared with the sham operation group， the model group showed broken and disordered myocardial fibers， cytoplasmic edema， and pyknosis and deviation of nuclei. Moreover， the modeling increased the levels of CK-MB and LDH （P<0.05， P<0.01）， lowered the levels of ATP， CCO， and SDH （P<0.05， P<0.01）， and down-regulated the mRNA and protein levels of SIRT1， PGC-1α， ERRα， and TFAM in myocardial tissue （P<0.05， P<0.01）. Compared with the model group， Wenxin prescription reduced the myocardial infarction area （especially in the high-dose group， P<0.01）， restored the pathological changes， lowered the levels of CK-MB and LDH （P<0.05， P<0.01）， increased the levels of ATP， CCO， and SDH （especially in the high-dose group， P<0.01）， and up-regulated the mRNA and protein levels of SIRT1， PGC-1α， ERRα， and TFAM in myocardial tissue （P<0.05， P<0.01）.ConclusionWenxin prescription can protect rats from myocardial ischemia-reperfusion injury by regulating myocardial mitochondrial energy metabolism via the SIRT1/PGC-1α/ERRα signaling pathway.
Keywords：Wenxin prescription;myocardial ischemia-reperfusion injury;mitochondrial energy metabolism;peroxisome proliferator-activated receptor-γ coactivator-1α （PGC-1α）
Abstract：ObjectiveTo explore the inhibitory effect of Bushen Jiedu prescription （BSJDP） on the metastasis of colorectal cancer （CRC） via activation of M2 tumor-associated macrophages （M2-TAMs） in vivo.MethodThe model of xenograft tumor was established with C57BL/6 mice， and then the model mice were randomly assigned into blank control group， Bushen Jiedu recipe-low dose （11.2 g·kg-1） group （BSJDP-L）， and Bushen Jiedu Recipe-high dose （22.4 g·kg-1） group （BSJDP-H）. Tumors were abolished when the volume reached 1.5-2.0 cm3. The mice were sacrificed 28 days post tumor abolishing and then the lung metastasis was observed. Histopathological changes in lung metastasis were examined by hematoxylin-eosin （HE） staining of metastasis tissues， and immunofluorescence （IF） staining was employed to observe the effect of BSJDP on tumor apoptosis and hypoxia. Flow cytometry was performed to analyze the macrophages M1/M2 ratio of tumor tissue. The expression levels of the genes （Arg1， CD206， and CD163） associated with the polarization of M2-TAMs were determined by Real-time fluorescent quantitative polymerase chain reaction （Real-time PCR）.ResultThe metastasis rate was 70%， 40%， and 10% in the blank control group， BSJDP-L group， and BSJDP-H group， respectively. The lower metastasis rates of BSJDP-L and BSJDP-H groups proved that BSJDP significantly inhibited lung metastasis of CRC. Compared with the blank control group， BSJDP-L and BSJDP-H reduced the tumor cell infiltration in tumor tissue （P<0.01）， increased the apoptosis of tumor cells， alleviated the hypoxic environment， and down-regulated the expression of Arg1， CD206， and CD163 in the tumor tissue （P<0.01）. In addition， the ratio of M2 macrophages ranked in a descending order of blank control group （34.867%） > BSJDP-L group （22.033%） > BSJDP-H group （11.633%） （P<0.01）.ConclusionBSJDP inhibits the lung metastasis of CRC by inhibiting the activation of M2-TAMs in the tumor microenvironment.
Abstract：ObjectiveTo investigate the effect of Mahonia bealei leaf extract on depression of rats and the underlying mechanism.MethodThe chemical constituents of the extract were analyzed by HPLC-MS/MS. Forced swimming test and tail suspension test were carried out to estimate the antidepressant effect. The mice were randomly assigned into the following groups： blank group， positive control group （fluoxetine， 10 mg·kg-1）， and Mahonia bealei leaf extract groups （10， 2.5 g·kg-1）. The gavage lasted for 12 days and the immobility time of the mice in the tests was recorded 1 h after the last administration. Furthermore， to explore the underlying mechanism of the antidepressant effect， we established the rat depression model by intraperitoneal injection with reserpine （0.5 mg·kg-1）. Rats were grouped as follows： blank group， model group， positive control group （fluoxetine， 1.8 mg·kg-1）， and Mahonia bealei leaf extract groups （10， 2.5 g·kg-1）. The gavage， once a day， lasted for 10 consecutive days. The depression of rats was detected by behavioral tests 1 h after the last administration. The levels of interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α） in serum of rats were detected by enzyme-linked immunosorbent assay （ELISA）. The protein expression of IL-6 and interleukin-1β （IL-1β） in brain tissue was detected by immunohistochemical staining. The protein levels of nuclear transcription factor-κB （NF-κB） and NOD-like receptor protein 3 （NLRP3） in hippocampus of rats were detected by Western blot.ResultSeven chemical constituents， mainly alkaloids， were identified from the extract. Compared with the blank group， Mahonia bealei leaf extract shortened the immobility time of mice in tail suspension and forced swimming tests. Compared with the blank group， the modeling of rat depression increased the blepharoptosis incidence and retention time in circles （P<0.05， P<0.01）， elevated the levels of IL-6 and TNF-α in serum （P<0.05）， and up-regulated the protein levels of IL-6， IL-1β， NF-κB， and NLRP3 in brain tissues （P<0.01）. Compared with the model group， high dose of Mahonia bealei leaf extract shortened the retention time in circles （P<0.05）， lowered the levels of IL-6 and TNF-α in serum （P<0.05， P<0.01）， and down-regulated the protein levels of IL-6， IL-1β， NF-κB， and NLRP3 （P<0.01） in brain tissues.ConclusionMahonia bealei leaf extract had significant antidepressant effect and alleviated the inflammatory response in reserpine-induced rat model of depression， the mechanism of which may be related to the inhibition of NF-κB/NLRP3 signaling pathway.
Keywords：Mahonia bealei leaves;depression;inflammation;NOD-like receptor protein 3 （NLRP3）;nuclear transcription factor-κB （NF-κB）
Abstract：ObjectiveSalidroside is the most abundant natural active compound in the famous Chinese herbal medicine Rhodiolae Crenulatae Radix et Rhizoma. This study aims to explore the effect of salidroside on the proliferation， migration， invasion， and apoptosis of human hepatoma （HepG2） cells.MethodThe HepG2 cells without any treatment were selected as the blank group， and the HepG2 cells in the salidroside groups were treated with salidroside at final concentrations of 20， 40， 80 μmol·L-1， respectively. A multifunctional cell analyzer， scratch assay， and Transwell assay were employed to determine the proliferation， migration， and invasion of HepG2 cells， respectively. An inverted microscope was used to observe the morphology， and a transmission electron microscope to observe the mitochondria of HepG2 cells. Flow cytometry was employed to determine the apoptosis and cycle distribution of HepG2 cells. Real-time fluorescent quantitative polymerase chain reaction ( Real-time PCR ) and Western blot were employed to determine the expression of apoptosis-associated genes and migration-， invasion-， and apoptosis-associated proteins， respectively， in HepG2 cells.ResultCompared with the blank group， salidroside （20， 40， 80 μmol·L-1） decreased the cell index and increased the healing area in a time- and dose-dependent manner （P<0.05）. Compared with that in the blank group， the HepG2 cells that could pass through Matrigel reduced in the salidroside （20， 80 μmol·L-1） groups. Compared with the blank group， salidroside （20， 40， 80 μmol·L-1） increased the total apoptosis rate in a dose dependence manner and blocked the cells in the G2/M phase （P<0.05）. Compared with the blank group， salidroside up-regulated the expression of epithelial-cadherin （E-cadherin） in a dose-dependent manner （P<0.05） and down-regulated that of nerve-cadherin （N-cadherin） in the 20 and 80 μmol·L-1 groups （P<0.05）. Compared with the blank group， salidroside （20， 40， 80 μmol·L-1） up-regulated the mRNA level of cysteine-containing aspartate-specific protease -3 （Caspase-3） and the protein levels of B-cell lymphoma-2 （Bcl-2） associated X protein （Bax）， Caspase-3， and cysteine-containing aspartate-specific protease-9 （Caspase-9） in a dose-dependent manner （P<0.05）， while it down-regulated the protein levels of the actin-binding protein Girdin and Bcl-2 in a dose-dependent manner （P<0.05）.ConclusionSalidroside inhibited the proliferation， migration， and invasion and induced the apoptosis of HepG2 cells through the mitochondrial pathway. The results suggest that salidroside can be used as a potential chemotherapy candidate for liver cancer.
Abstract：ObjectiveTo explore the effect of Jinshui Xiangsheng prescription on the five-year clinical survival outcome of patients with advanced prostate cancer.MethodFrom May 1, 2014 to May 1, 2016, patients with advanced prostate cancer from Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine and the Urology Department of the Second Affiliated Hospital of Nanjing University of Chinese Medicine were collected and treated with Jinshui Xiangsheng prescription (155 cases in the observation group). According to age and Gleason score, the patients without Jinshui Xiangsheng prescription were matched in a ratio of 1∶1 (155 cases in the control group). The androgen resistance rate, survival rate, median survival time, and median progress free survival time in 1, 3, 5 years were observed. The prognostic factors of advanced prostate cancer were analyzed and screened out by Chi-square test, t test, Kaplan-Meier and Cox survival analysis.ResultThe androgen resistance rates in the observation group in 1， 3， 5 years were 9.0% （14/155）， 72.3% （112/155）， and 92.9% （144/155）， respectively， and those in the control group were 20.6% （32/155）， 87.7% （136/155）， and 97.4% （151/155）. The 1-year （χ2=8.271，P<0.01）and 3-year （χ2=11.613，P<0.01） androgen resistance rates in the observation group were significantly lower than those in the control group. The median survival time and median progress free survival time in the observation group were （26.35±9.01） months and （11.02±4.40） months， respectively， and in the control group were （22.31±9.21） months and （9.87±5.12） months， respectively. The median survival time and median progress free survival time in the observation group were significantly longer than those in the control group （P<0.05， P<0.01）. The cumulative survival rates in 1， 3， 5 years in the observation group were 96.1% （149/155）， 80.6% （125/155）， and 39.4% （61/155）， respectively， and those in the control group were 94.2% （146/155）， 60.0% （93/155）， and 22.6% （35/155）， respectively. The 3-year （χ2=15.828，P<0.01） and 5-year （χ2=10.201，P<0.01） cumulative survival rates in the observation group were significantly higher than those in the control group. The monofactor analysis showed that the prognostic factors involved in Gleason score， initial prostate specific antigen （PSA）， tumor location， tumor stage， castration regimen， radiotherapy， chemotherapy， complete androgen blockade （CAB）， and Jinshui Xiangsheng prescription （P<0.05， P<0.01）. The multivariate analysis showed that initial PSA， tumor location， and tumor stage were the risk factors affecting the survival time of patients with advanced prostate cancer， whereas Jinshui Xiangsheng prescription， castration regimen， chemotherapy， radiotherapy， and CAB were the independent protective factors affecting the prognosis of advanced prostate cancer.ConclusionJinshui Xiangsheng prescription has a protective effect on the survival of patients with advanced prostate cancer， which reduces the androgen resistance rate and death risk of advanced prostate cancer， thus benefiting the survival of patients. Therefore， it deserves further promotion.
Abstract：ObjectiveTo study the effect on quality of life and the bone turnover markers of Buzhong Yiqitang in the treatment of senile osteoporotic vertebral compression fracture （OVCF， syndrome of Qi deficiency in spleen and stomach） after operation based on ''spleen governing muscle''.MethodA total of 135 senile patients with OVCF treated by percutaneous kyphoplasty in Rizhao Hospital of Traditional Chinese Medicine from January 2020 to January 2021 were enrolled in this study. They were randomly assigned to two groups on the basis of block randomization at a ratio of 2∶1 （90 cases in the observation group and 45 cases in the control group）. Both groups were administrated with calcitriol capsules （0.5 μg·d-1） and caltrate D （1 200 mg·d-1） for basic treatment of osteoporosis. The observation group was additionally treated with Buzhong Yiqitang. Bone mineral density （BMD）， procollagen type Ⅰ N-terminal propeptide （PINP）， osteocalcin （OST）， β cross-linked C-telopeptide of type 1 collagen （β-CTx）， appendicular skeletal muscle mass index （ASMI）， and quadriceps muscle strength were compared between the two groups before and 6， 12 months after treatment. Additionally， traditional Chinese medicine （TCM） symptom score and visual analogue score （VAS） before and 3， 6 months after treatment， as well as quality of life questionnaire of the European Foundation for osteoporosis （QUALEFFO） score before and 3， 6， 12 months after treatment， were compared between the two groups.ResultA total of 85 patients in the observation group and 41 patients in the control group were followed up. The general curative effect of the observation group was better than that of the control group （χ2=10.503， P<0.05）. Specifically， the observation group had higher PINP， BMD， ASMI， and quadriceps muscle strength but lower β-CTx， TCM symptom score， VAS， and QUALEFFO score than the control group （P<0.05， P<0.01）. No adverse reactions related to Buzhong Yiqitang were observed.ConclusionBuzhong Yiqitang can regulate bone metabolism indexes， promote osteogenesis， increase bone density， enhance skeleton appendiculare and quadriceps muscle strength， relieve clinical symptoms， and improve quality of life in patients with senile OVCF （syndrome of Qi deficiency in spleen and stomach）， being worthy of promotion in clinical application.
Keywords：Buzhong Yiqitang;senile osteoporotic vertebral compression fracture;bone turnover markers;quality of life
Abstract：ObjectiveTo study the clinical efficacy of dapagliflozin combined with Shexiang Baoxinwan （SXBXW） in the treatment of acute heart failure with reduced ejection fraction （HFrEF） and syndrome of Qi deficiency and blood stasis.MethodA total of 176 patients hospitalized due to acute HFrEF （syndrome of Qi deficiency and blood stasis） were selected and randomized into control group， SXBXW group， dapagliflozin group， and SXBXW + dapagliflozin group （the latter three groups were called the intervention groups）. The New York Heart Association （NYHA） class， 6-minute walk test （6MWT） score， Kansas City Cardiomyopathy Questionnaire （KCCQ） score， traditional Chinese medicine （TCM） syndrome score， N-terminal pro-brain natriuretic peptide （NT-proBNP）， soluble suppression of tumorigenicity 2 （sST2）， interleukin-6 （IL-6）， and hypersensitive C-reactive protein （hs-CRP） of the patients were evaluated and measured at the time of admission， 1 week after treatment， and 2 weeks of treatment. Furthermore， the hospital stay， in-hospital mortality， and 30-day re-admission rate were recorded. Statistical analysis was performed to evaluate the efficacy of each group.ResultAfter 1 week of treatment， the SXBXW group exhibited superior NYHA class， KCCQ score， TCM syndrome score and curative effect， IL-6， and hs-CRP to the control group （P<0.05， P<0.01）. After 2 weeks of treatment， the SXBXW group showed superior TCM syndrome score， TCM curative effect， and hs-CRP （P<0.05， P<0.01） to the control group. The dapagliflozin group was superior to the control group in terms of TCM syndrome score， NT-proBNP， and sST2 （P<0.05， P<0.01） after 1 week of treatment and in terms of NYHA class， KCCQ score， NT-proBNP， sST2， and hospital stay （P<0.05， P<0.01） after 2 weeks of treatment. The SXBXW + dapagliflozin group exhibited better efficacy than the control group in terms of NYHA class， 6MWT score， KCCQ score， TCM syndrome score and curative effect， NT-proBNP， sST2， IL-6， and hs-CRP （P=0.014） after 1 week of treatment and in terms of NYHA class， KCCQ score， TCM syndrome score and curative effect， NT-proBNP， sST2， IL-6， hs-CRP， and hospital stay （P<0.01） after 2 weeks of treatment.ConclusionSXBXW and dapagliflozin have good therapeutic effect on acute HFrEF and syndrome of Qi deficiency and blood stasis， and their combination demonstrated better therapeutic effect， with good safety and tolerability.
Keywords：heart failure with reduced ejection fraction;syndrome of Qi deficiency and blood stasis;acute attack;dapagliflozin;Shexiang Baoxinwan;clinical efficacy
Abstract：ObjectiveTo establish a polymerase chain reaction（PCR） method to accurately discriminate the crude materials of Murrayae Folium et Cacumen， Murraya exotica and M. paniculata.MethodBased on the difference in chloroplast genome sequences of M. exotica and M. paniculata， species-specific identification primers P03 and P04 of M. exotica and M. paniculata were designed according to single nucleotide polymorphism （SNP） on the chloroplast genome. A multiplex allele-specific PCR identification method was established for the identification of M. exotica and M. paniculata following the optimization of annealing temperature， number of cycles， and primer concentration ratio. The established PCR method for identification was explored and verified in terms of tolerance and feasibility by investigating the type of Taq polymerases and PCR system model.ResultIn this multiplex allele-specific PCR identification method， about 330 and 230 bp of specific fragments were amplified from DNA templates of M. exotica and M. paniculata， respectively， under the following conditions：cycle number of 31， annealing temperature of 60 ℃， and primer concentration ratio of P03 and P04 of 1∶2. Consistent results were obtained for samples from different sources.ConclusionThe multiplex allele-specific PCR identification method established in this study can accurately identify the origin of Murrayae Folium et Cacumen， which can be used for the simultaneous identification of M. exotica and M. paniculata by the length of fragments in a single identification assay.
Abstract：ObjectiveTo establish a rapid screening method for germplasm materials of Gastrodia elata with high purity， and lay a foundation for pure line breeding and cross breeding.MethodBased on the whole genome sequencing and population resequencing of G. elata， 20 restriction fragment length polymorphism (RFLP) markers were developed by single nucleotide polymorphism (SNP) sites. The polymerase chain reaction (PCR)-RFLP method was used to carry out restriction endonuclease experiments on 20 RFLP markers of 15 G. elata germplasms. According to the number of enzymatic bands at 20 RFLP marker sites, the purity of 15 germplasms was calculated and evaluated. On this basis, genome resequencing technology was used to verify the assessment results.ResultTen germplasm materials with purity greater than 95% were screened out by PCR-RFLP method， 3 of which had 95% purity and 7 had 100% purity. Nine germplasm materials with purity greater than 95% were screened out by genome resequencing methods, and 8 of them were consistent with the results of PCR-RFLP.ConclusionThe PCR-RFLP method established in this study for screening G. elata germplasms with high purity precision of RFLP markers has 80% precision and 89% accuracy. The method is simple， efficient， and significantly less expensive than genome resequencing method, which provides technical support for pure line breeding of G. elata and references for breeding of other Chinese medicinal materials.
Abstract：ObjectiveIn order to ensure the safety and effectiveness of clinical drug use ， the identification method of mixed and adulterated specific polymerase chain reaction （PCR） identification of Pheretima aspergillum and its processed products was established.MethodBased on the cytochrome C oxidase subunit I sequence of P. aspergillum， primers were designed to cover the whole sequences， and the stable DNA ranges suitable for the identification of Pheretima （P. aspergillum） formula granule were screened out. Specific primers were designed according to the specific single nucleotide polymorphisms （SNP） of P. aspergillum in the stable DNA range. The P. aspergillum and its mixture were collected respectively， the PCR reaction system was established and optimized， and PCR reaction system and procedure were optimized， and the tolerance and applicability were investigated.ResultWhen the annealing temperature was 62 ℃ and the cycle number was 36， both P. aspergillum formula granule and its formula particles could amplify a single specific identification band of about 170 bp， and the other 20 adulterants and negative controls had no band.ConclusionThe allele-specific PCR identification method established in this study can quickly and accurately identify the P. aspergillum formula granule. The orgin of Chinese herbal medicine and decoction pieces and P. aspergillum were accurately identified. It can also provide a reference for other studies on the quality standard research of other Chinese herbal formula granule.
Abstract：ObjectiveTo establish a specific polymerase chain reaction （PCR） method for the identification of Artemisia absinthium to allow accurate and convenient identification of A. absinthium and its related species.MethodThe chloroplast genome sequences of A. absinthium and its related species were searched from Chloroplast Genome Information Resource （CGIR）， and the specific single nucleotide polymorphism （SNP） sites of A. absinthium were screened out. A pair of specific identification primers （zykh1-F and zykh1-R） of A. absinthium was designed. The original plant samples of A. absinthium and its related species were collected. The specific PCR method was established and optimized， and the tolerance and feasibility of this method were investigated and verified. The method was used to identify A. absinthium samples purchased from Xinjiang medicinal materials market.ResultA 210 bp bright band was obtained from A. absinthium after PCR amplification and gel electrophoresis under the following conditions： specific primers zykh1-F and zykh1-R， annealing temperature of 54 ℃， and the number of cycles of 33. No such band was observed from its relative species， such as A. argyi， A. annua， A. leucophylla， and A. lavandulaefolia.ConclusionThe specific PCR identification method of established in this study can accurately identify A. absinthium and its common related species with high specificity. The method can save time and cost and allows a convenient and fast species identification for the introduction and utilization of A. absinthium resources.
Abstract：ObjectiveUncommon medicinal herbs are valuable medicinal resources， but their identification is a difficult problem in Chinese medicine due to their particularity and complexity. It is， therefore， urgent to establish a method for the identification of uncommon medicinal herbs. In this study， DNA signature sequence （DSS） tags were used to establish a specific polymerase chain reaction （PCR） identification method for Hibisci Cortex， the origin plant of Hibisci Cortex， and its adulterants.MethodThe candidate DSS tags were obtained from the chloroplast genome sequence analysis， and the DSS tags were verified by DNA sequencing. The specific identification primers for H. syriacus were designed based on the obtained reliable DSS tags. The PCR reaction conditions were optimized， and the tolerance and feasibility were investigated.ResultA DSS tag for identification of H. syriacus was obtained from the comparison of sequencing results of the amplified products with DSS， which revealed the distinguishing characteristics of Hibisci Cortex and its adulterants. A pair of specific primers for H. syriacus was designed according to the DSS tag. After PCR amplification and gel electrophoresis with the primers， a single bright band of about 270 bp was observed from H. syriacus， which did not appear in the four adulterants.ConclusionA DSS tag obtained in this study can be used to identify H. syriacus. The specific primers designed based on this DSS tag can accurately and simply identify the original plant of Hibisci Cortex and its adulterants， which provides a new method and idea for the molecular identification of genuine and counterfeit products of Hibisci Cortex.
Abstract：ObjectiveTo clarify the spatial distribution characteristics of medicinal plant resources in Gansu province， analyze the causes， changing trends， and driving factors of the spatial differentiation， and thus lay a scientific basis for the rational development and sustainable development of medicinal plant resources in this province.MethodBased on the data of The Fourth National Survey of Chinese Medicine Resources， the richness and spatial distribution difference of medicinal plant resources in 87 counties （districts） of Gansu province were analyzed via the global spatial autocorrelation analysis， trend surface analysis， local spatial autocorrelation analysis， and hotspot analysis. Moreover， the correlation of vegetation type， soil texture， annual average temperature， annual average precipitation， and altitude with the spatial distribution pattern of the medicinal plant resources was discussed.ResultCounties （districts） with high or low richness of medicinal plant resources in Gansu province were respectively clustered together. To be specific， counties （districts） with high richness of the medicinal resources were mainly in southeastern Gansu， while those with low richness in northwestern Gansu. The leading driving factors affecting the cold and hot spots included vegetation type， soil texture， and average annual rainfall.ConclusionThe species richness of medicinal plant resources in Gansu province rises from west to east and from north to south. The natural driving factors are the key to the diversity and spatial distribution pattern of medicinal plant resources， which show significant influence on them.
Keywords：The Fourth national Survey of Chinese Medicine Resources;spatial distribution;medicinal plants;species richness;natural driving factor
Abstract：ObjectiveIn order to solve the problem that the quality and stability of Arisaema Cum Bile in the fermentation process with hybrid bacteria were not easy to control， the microorganism in the fermentation process of Arisaema Cum Bile was isolated and identified， the dominant strains were screened and the fermentation process of Arisaema Cum Bile with compound bacteria was investigated.MethodThe submerged culture during the fermentation process of Arisaema Cum Bile was taken out for strain separation and purification. Bacteria and fungi multiphase identification and detection methods and automatic microbial analysis system were used to analyze and compare DNA sequences and identify microorganisms. The isolated and identified strains were respectively inoculated and fermented. After screening the dominant strains， a preliminary exploration of compound strain fermentation were carried out. The contents of index components in Arisaema Cum Bile fermented by compound strain and traditional Arisaema Cum Bile were compared by ultra-performance liquid chromatography-triple quadrupole tandem mass spectrometry （UPLC-QqQ-MS/MS）. Mmobile phase was 0.1% formic acid acetonitrile solution （A）-0.1% formic acid aqueous solution （B） for gradient elution （0-2 min， 35%-45%A； 2-10 min， 45%-48%A； 10-12 min， 48%-100%A； 12-12.01 min， 100%-35%A； 12.01-15 min， 35%-65%A）， the flow rate was set at 0.35 mL·min-1. The mass spectrographic analysis employed electrospray ionization （ESI）， negative ion acquisition mode and multiple reaction monitoring （MRM） scanning mode were adopted to collect information， the collection range was m/z 50-1 000.ResultEight microorganisms were isolated and identified from the submerged culture of Arisaema Cum Bile. Among them， Enterococcus sp. （anaerobic） and E. casseliflavus were selected as the dominant strains in the fermentation process. Compared with the traditional fermentation method， the contents of chenodeoxycholic acid， hyodeoxycholic acid and hyocholic acid in free cholic acid increased by 1.76， 0.06， 0.19 mg·g-1， respectively. In bound cholic acid， glycochenodeoxycholic acid， taurochenodeoxycholic acid， glycohyodeoxycholic acid， taurohyodeoxycholic acid， glycohyocholic acid， taurine porcine cholic acid decreased by 0.63， 0.23， 0.26， 0.16， 0.03， 0.04 mg·g-1， respectively.ConclusionArisaema Cum Bile with compound strain fermentation （Enterococcus sp. and E. casseliflavus） can be fermented more completely， the fermentation cycle can be shortened， and the quality and stability of products can be improved.
Keywords：Arisaema cum Bile;fermentation process;strain identification;traditional Chinese medicine processing;dominant strain;bile acid;ultra-performance liquid chromatography-triple quadrupole tandem mass spectrometry （UPLC-QqQ-MS/MS）
Abstract：ObjectiveThe internal transcribed spacer （ITS） 2 region of ribosomal gene， a DNA barcode， was employed to identify 12 medicinal Aconitum species and the genetic relationship among the species was analyzed.MethodA total of 30 samples of the 12 species were collected. The DNA was extracted with spin column plant genomic DNA kit and the universal primers of ITS2 sequence were used for polymerase chain reaction （PCR） amplification， followed by electrophoresis detection and bi-directional sequencing. The yielded sequences were aligned and spliced by CodonCode Aligner 17.0 and sequence variation was analyzed by MEGA 7.0. The secondary structure was predicted by ITS2 Database and the neighbor-joining （NJ） method was applied to generate the phylogenetic tree.ResultThe ITS2 sequences of the 12 species were 220-221 bp， with the average guanine and cytosine （GC） content of 64.09%， 140 variable sites， 137 informative sites， and 81 conservative sites. The intraspecific genetic distance （K2P） was smaller than the interspecific genetic distance. According to the secondary structures of ITS2 sequences and NJ cluster analysis， A. scaposum， A. sinomontanum， and A. barbatum had close genetic relationship， while the rest nine showed close kinship， particularly A. soongaricum and A. yinschanicum.ConclusionITS2 sequence is of great value for the molecular identification and genetic relationship determination of Aconitum， which provides a new method for the study of ethnomedicine.
Abstract：ObjectiveTo predict the underlying mechanism of Bushen Huoxuetang in treating osteoporosis related to endocrine therapy in breast cancer by network pharmacology and to verify the results through in vitro cell model.MethodThe main effective components and targets of Bushen Huoxuetang were screened out through network pharmacology， and the targets of osteoporosis related to endocrine therapy in breast cancer were further obtained. The intersected targets were analyzed by Gene Ontology （GO） functional annotation and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment. Kaplan Meier plotter was used to analyze the survival of crucial targets. Finally， the inhibitory activity against cell proliferation was evaluated by in vitro methye thiazolye telrazlium（MTT） assay. The key targets and pathways were verified by Western blot， and the mRNA expression of the key targets was evaluated by real-time polymerase chain reaction（Real-time PCR）.ResultA total of 716 active components and 249 key targets of Bushen Huoxuetang were obtained from network pharmacology. There were 135 common targets， among which protein kinase B（Akt）1 and hypoxia-inducible factor-1α （HIF-1α） were two key targets. Additionally， 531 biological processes， 62 cellular components， 162 molecular functions， and 145 signaling pathways including breast cancer and endocrine resistance were involved. The key targets were effectively enriched in phosphatidylinositol 3-kinases（PI3K）/Akt and HIF-1 signaling pathways. According to the MTT assay， the cell proliferation rate and cell motility of MCF-7 and T47D cells in the luminal A cell line were reduced by Bushen Huoxuetang treatment （22.5， 45， 90 g·L-1， and 45， 90， 180 g·L-1） for 48 h as compared with the blank group. As revealed by Western blot， MCF-7 cells were treated with Bushen Huoxuetang （0， 15， 60 g·L-1） for 48 h， and the relative expression of p-PI3K， PI3K， p-Akt， Akt， and HIF-1α was decreased in a dose-dependent manner as compared with the blank group （P<0.05， P<0.01）. Real-time PCR was used to detect the mRNA expression of the key target HIF-1α. The results showed that the mRNA expression of HIF-1α in MCF-7 cells was decreased with the increase in the dose （P<0.01）， and the change was in a concentration-dependent manner.ConclusionThe mechanism of Bushen Huoxuetang in the treatment of osteoporosis related to endocrine therapy in breast cancer may be related to the key targets including Akt1 and HIF-1α through the PI3K/Akt/HIF-1α signaling pathway.
Abstract：ObjectiveEsophageal cancer is a common malignant tumor of the upper gastrointestinal tract， and has high incidence and mortality in China. Its incidence is increasing year by year， and survival rate is low， thus seriously threatening human life and health. To further explore the pathogenesis of esophageal cancer and its systematic and efficient diagnosis and treatment methods， the animal models of esophageal cancer was evaluated according to the animal model evaluation method previously established by our team based on the characteristics of its clinical symptoms of traditional Chinese and western medicine， and suggestions for model improvement were proposed.MethodThe existing animal models of esophageal cancer were summarized through China National Knowledge Infrastructure （CNKI） and Wanfang Data. The relevant indexes of the models were assigned， and their coincidence with the clinical diagnostic guidelines of traditional Chinese and western medicine for esophageal cancer was evaluated.ResultExcept the spontaneous animal model of esophageal cancer with high clinical coincidence adopted in few studies， the animal model induced by methylbenzylnitrosamine was in good agreement with the clinical data， which simulated the etiology and pathogenesis of esophageal cancer to a certain extent. The model partially reflected some indicators of clinical diagnosis in western medicine， and also indicated the body weight loss， purple nail and increased number of drinking in traditional Chinese medicine （TCM）. However， there was still a lack of differentiation of TCM syndromes.ConclusionOn the basis of the original model， the animal model induced by methylbenzylnitrosamine and the mouse model of xenogeneic gastric wall transplantation of human esophageal cancer cells were applied， which were intervened with the factors of phlegm and qi mutual obstruction syndrome， blood stasis and phlegm stagnation syndrome， Yin deficiency and internal heat syndrome and Qi deficiency and yang weakness syndrome， and were distinguished to reflect the performance of TCM syndrome. The animal model of esophageal cancer combined with TCM syndrome was thus obtained， which embodied the clinical symptoms of esophageal cancer in TCM， and the characteristics of the animal model combined with TCM syndrome， and simulated the clinical practice of traditional Chinese and western medicine， providing an accurate pathological model carrier for basic research.
Keywords：esophageal cancer;clinical characteristics of disease and syndrome;animal model;evaluation analysis
Abstract：Alzheimer's disease （AD） is a neurological disease highly related to age， which is the main cause of senile dementia and the most common disease leading to the loss of daily living ability of the elderly. AD brings heavy mental burden and economic pressure to patients， families， and society. Traditional Chinese medicine （TCM） ascribes AD to category of "dementia"， believing that the treatment should start from kidney because kidney deficiency is the root cause. Combined with the physiological and pathological characteristics of liver， this paper proposed that liver-kidney homology was an important idea for the prevention and treatment of AD. The main pathological manifestations of AD were amyloid β-protein （Aβ） deposition and neurofibrillary tangles （NFT）， and the pathogenesis was complex. A growing number of studies showed that immune inflammation played an important role in the pathogenesis of AD. The important target of treating AD was the regulation of neuro-immune inflammation through the nuclear factor kappa B （NF-κB）/NOD-like receptor thermal protein domain associated protein 3 （NLRP3）/Caspase-1/interleukin-1β （IL-1β） signaling pathway. Based on the idea of liver-kidney homology， this paper selected the representative formula Hei Xiaoyaosan to explore its effect on the prevention and treatment of AD and the mechanism from the perspective of regulating NF-κB/NLRP3/Caspase-1/IL-1β signaling pathway and inhibiting neuro-immune inflammation， expecting to further promote the in-depth study on the prevention and treatment of AD， and provide references for the prevention and treatment of AD by TCM.
Abstract：After the completion of the pilot work of traditional Chinese medicine formula granules （TCMFGs）， the national and provincial medical products administrations have published and implemented about 440 varieties of TCMFGs standards. Based on the previous work， this paper analyzed technical problems encountered in the review and evaluation of Shandong TCMFGs standards， mainly involving the executive standards and distinguishing technologies of raw materials， the adding process of excipients in the procedure item， the rationality of quality control methods， the information content and reproducibility of characteristic chromatograms， the nomenclature and accessibility of reference materials， etc. The common problems such as the coverage of standards， specification differences， and the integrity of quality control items of current TCMFGs standards were discussed deeply. It is proposed to promote the upgrading of provincial standards to national standards， accelerate the research and development of reference materials， advocate the use of high-quality raw materials， explore the evaluation methods of high-quality products， and strengthen the quality supervision of the whole process. Suggestions of this paper is hoped to provide references for the formulation of national and provincial TCMFGs standards， promote the continuous improvement of TCMFGs standard system， and ensure the healthy and orderly development of the TCMFGs industry.
Keywords：traditional Chinese medicine formula granules;standard formulation;quality control;production technology;whole process supervision;technical problems;countermeasure analysis
Abstract：Cardiovascular diseases are the leading cause of death in the world today. Atherosclerosis （AS） is a chronic inflammatory disease characterized by thickening or functional degeneration of the arterial wall， and in the later stage of the disease， plaque ruptures to induce thrombosis， which in turn causes ischemia in tissues or organs. It is therefore the pathological basis for all types of cardiovascular diseases. Nuclear transcription factor kappa B （NF-κB）， an important nuclear transcription factor in the inflammatory response， is activated to mediate the transcription of inflammatory factors that can trigger or exacerbate the development of AS. Vascular endothelial cells are activated by inflammatory factors. NF-κB mediates related regulatory genes in endothelial cells to secrete adhesion molecules， chemokines， and coagulation factors， promotes selective aggregation of monocytes， up-regulates the expression of adhesion molecules to make adhesion molecules stick to the endothelium and move toward the intima， promotes the degradation of the extracellular matrix， and forms unstable plaques. In recent years， traditional Chinese medicine （TCM） has achieved certain results in the prevention and treatment of AS， and many Chinese medicines have been proved to be effective in resisting AS and can act on multiple targets in the human body， affecting the occurrence and development of AS in different links. This paper mainly introduced the NF-κB pathway and its relationship with AS， reviewed research progress on 75 components of different types in Chinese medicine monomers such as flavonoids， terpenoids， and alkaloids in AS resistance based on the NF-κB pathway， and found that Chinese medicine monomers mainly regulate cholesterol balance， inhibit the inflammatory response， reduce cell proliferation， inhibit intercellular adhesion， and suppress foam cell formation by regulating the NF-κB pathway to provide a reference for the prevention and treatment of AS.
Keywords：atherosclerosis;nuclear transcription factor-kappa B （NF-κB）;Chinese medicine;inflammation;Chinese medicine monomer
Abstract：Paclitaxel is the first-line chemotherapy drug for a variety of cancers. However， the paclitaxel resistance greatly reduced the efficacy in the later treatment stage， which seriously increased the mortality and recurrence rate of cancer and limited the clinical application of paclitaxel. At present， Chinese medicine compound prescription， proprietary Chinese medicine， and Chinese medicine injection are widely used as the adjuvant chemotherapy drugs for the treatment of cancer in clinic. Chinese medicine has shown unique advantages in improving the efficacy of chemotherapy drugs and the prognosis of chemotherapy， and reducing the toxic and side effects. However， the specific mechanism and effective monomer composition of Chinese medicine for reversing the resistance of chemotherapy drugs are unclear， and the application of Chinese medicine in different types of cancer is also limited， which are worthy of further exploration. This review summarized the composition of Chinese medicine monomer with synergistic antitumor effect combined with paclitaxel in recent years. The specific mechanism and pharmacological activities of Chinese medicine monomer reversing paclitaxel resistance were classified. This review found that through acting on the membrane transport protein， Chinese medicine monomer promoted the accumulation of paclitaxel in tumor cells， inhibited the expressions of protein and metabolic enzyme related to multidrug resistance and the metabolism of paclitaxel， and regulated the levels of apoptosis genes and factors and apoptosis-related pathways to promote the inhibitory effect of paclitaxel on cell proliferation. Chinese medicine monomer also significantly improved paclitaxel chemotherapy sensitivity by regulating the expression levels of micro ribonucleic acid （microRNA） and long non-coding ribonucleic acid RNA （lncRNA）， inhibiting the characteristics of tumor stem cells and tumor metabolic reprogramming， improving tumor microenvironment， and triggering tumor cell death autophagy and oxidative stress response. This review provides a theoretical basis for clarifying the specific anti-tumor mechanism of Chinese medicine monomer combined with paclitaxel， which is of great significance for the development of new Chinese medicine and the clinical research of the drugs combined with paclitaxel， and has certain value for the application of Chinese medicine combined with other chemotherapy drugs.
Keywords：Chinese medicine monomers;paclitaxel;drug combination;mechanism of reversing drug resistance
Abstract：Liver cancer is a worldwide malignant tumor with an increasing incidence by years. At present， it is facing the predicament of poor prognosis and lack of effective therapeutic drugs. Epimedii Folium is a well-known traditional Chinese medicine with a long history， and exiting clinical and pharmacological studies show that it can be used in clinical treatment of liver cancer. According to reports， Epimedii Folium polysaccharides （EPS）， C-8-isopentenyl substituted flavonoids and their glycosides （icaritin， icariin， baohuoside Ⅰ， epimedin C） have good anti-liver cancer activity. They are the main active ingredients of Epimedii Folium against liver cancer. The data which comes from in vitro and in vivo studies suggests flavonoids in Epimedii Folium demonstrate anti-liver cancer activity through various mechanisms， including inhibiting hepatoma cells proliferation， promoting hepatoma cells apoptosis， improving tumor immunosuppression microenvironment， inhibiting hepatoma cells immune escape， invasion and migration， reversing hepatoma cells resistance， suppressing hepatocellular carcinoma initiation cells and regulating the immunity of the body. While EPS play an anti-hepatocellular carcinoma role mainly through the regulation of immunity. Epimedii Folium exerts good anti-liver cancer effects with multiple components， multiple targets， and multiple pathways， which makes it a valuable anti-liver cancer drug. However， the comprehensive analysis of related aspects is still lacking. Therefore， this study briefly reviewed the anti-hepatocellular carcinoma active ingredients of Epimedii Folium and their mechanisms. In addition， in the process of literature review， it was found that the anti-liver cancer studies of Epimedii Folium mainly focused on a few components and the studies elucidating the active constituents and mechanism of Epimedii Folium against liver cancer on the whole level were insufficient. Based on these questions， the study also proposed corresponding suggestion to provide reference for the further study of substance basis， clinical application and rational development of Epimedii Folium against liver cancer.
Abstract：Breast cancer is a systemic malignant tumor caused by multiple pathogenic factors， and its pathological mechanism is complex and has not been clarified so far. It has gradually become the largest killer threatening women's life. The common method for the treatment of breast cancer is lesion resection combined with radiation and chemical therapy， endocrine therapy， or targeted therapy. However， due to the limitations of western medicine therapies， there are still considerable breast cancer patients with poor disease control and high tumor recurrence rate in clinical practice. At the same time， the side effects and complications produced by these therapies affect the quality of life of patients. Therefore， it is urgent to develop new drugs or find safe and effective alternative therapies against breast cancer. Volatile oil （VO）， as a unique volatile component of Chinese herbal medicines， has anti-inflammatory， antiviral， and anti-tumor activities. It has been applied in the treatment of breast cancer and has demonstrated good efficacy by exerting the unique effects of strengthening healthy Qi， eliminating pathogenic factors， moving Qi， resolving stasis， warming Yang， soothing liver， and relieving depression. The recent studies have confirmed that VO and its chemical components can prevent and treat breast cancer via multiple mechanisms， while there is a lack of systematic review. The relevant literature published in recent years has demonstrated that VO can inhibit the occurrence and development of breast cancer by regulating the level of estrogen， inhibiting the proliferation and inducing the apoptosis of cancer cells， enhancing immunity， resisting inflammation， and regulating emotions. We introduced the pathogenesis of breast cancer， as well as the mechanisms and advantages of VO in the prevention and treatment of breast cancer， aiming to provide new ideas for the research on VO in the treatment of breast cancer.
Keywords：volatile oil from Chinese medicine;breast cancer;etiology and pathogenesis;advantages;mechanism of action
Abstract：In China， the incidence of ischemic heart disease （IHD） is increasing year by year， which brings enormous burden to families and society. It is urgent to find preferable treatment methods and medical therapies. The Chinese ethnic minority medicine has gradually developed unique theoretical systems and therapeutic characteristics on the basis of clinical experience and thinking modes including image-number thinking and the holistic perspective. Consequently， it possesses huge application capacity and research value in prevention and treatment of IHD. Belonging to the medical system based on the view of nature and life， the Tibetan medicine， Mongolian medicine， and Dai medicine have respectively formed theories like "three elements" "three life-sustaining energies" "four elements and five skandhas （aggregates）" ， have put forward unique understandings of IHD and have formed corresponding therapeutic principles and methods， generating plentiful classic prescriptions represented by Sanwei Tanxiang powder， Bawei Chenxiang powder， Roukou Wuwei pills and Yajiao Hadun powder. They also contain characteristic ethnic medicine resources such as Choerospondiatis Fructus， Rhodiola Rosea and Draconis Sanguis. Aiming to provide enlightenment and reference for the clinical application and development of the Chinese ethnic minority medicine for the prevention and treatment of IHD， the authors try to summarize the related researches represented by Tibetan and Mongolian medicines， and then discuss the opportunities and challenges faced by such researches.
Keywords：ethnic medicine;ischemic heart disease;Tibetan medicine;Mongolian medicine;Dai medicine;basic research;clinical research
Abstract：Cerebral ischemia-reperfusion injury （CIRI） is a further injury of brain tissue after the recovery of blood supply in ischemic stroke， which seriously affects the quality of life of patients. The pathological mechanism of CIRI is complex， mainly involving excitatory amino acid toxicity， apoptosis， inflammation， and oxidative stress. Studies have proved that Chinese herbal medicines and their active components have unique advantages and good application prospects in the prevention and treatment of CIRI. Quercetin is a flavonoid ubiquitous in a variety of Chinese medicinal herbs. It can alleviate CIRI and reduce brain injury through inhibiting inflammation， oxidation， and apoptosis， protecting blood-brain barrier， and activating mitophagy. However， little is known about the specific mechanism and molecular targets. In view of the low bioavailability and poor solubility of quercetin， researchers have developed a variety of delivery systems to facilitate the dispersion of quercetin， improve chemical stability， and increase clinical application. Furthermore， researchers have tested the long-term safety of quercetin and confirmed that low-dose quercetin has good safety. By reviewing the relevant studies in recent years， we summarized the targets， mechanism， delivery， and safety of quercetin in the treatment of CIRI， aiming to provide a theoretical basis for the further development and application of quercetin.
Keywords：quercetin;cerebral ischemia-reperfusion injury;mechanisms of action;drug delivery;safety evaluation
Abstract：As of 2020， there are more than 120 million diabetic patients in China. Diabetic wounds is one of the common complications of diabetes with increasing incidence and has the potential to cause disability and mortality. Traditional Chinese medicine （TCM） has a long history in treating diabetic wounds， demonstrating significant efficacy and safety. In recent years， increasing researchers have explored the mechanisms of polysaccharides from TCM in the repair of diabetic wounds. Polysaccharides are the main active ingredients of TCM and employ one or more blood sugar-lowering mechanisms. However， most studies focus on the repair mechanism of single polysaccharides， and there is little in-depth discussion and summary. To provide a new therapy for diabetic wounds， which meets international standards and has the characteristics of TCM， and provide reference for the clinical treatment of diabetic wounds， we reviewed relevant literature to summarize the mechanisms of TCM polysaccharides in treating diabetic wounds. The mechanisms include inhibiting inflammation to improve wound microenvironment， lowering blood sugar， promoting fibroblast migration and proliferation， regulating wound growth factor to promote angiogenesis， inhibiting oxidative stress response， and regulating immune function. Finally， we put forward some possible research directions in the future.
Keywords：diabetes mellitus;diabetic wound;polysaccharides from traditional Chinese medicine;mechanism of action;research progress
Abstract：Citri Reticulatae Pericarpium is a commonly used traditional Chinese medicine in China. By referring to herbal monographs and related research documents， and comparing all editions of Chinese Pharmacopoeia and local processing specifications， the processing history， modern processing specifications， study on chemical constituents and pharmacological effects before and after processing of Citri Reticulatae Pericarpium were summarized. It was found that the processing methods of Citri Reticulatae Pericarpium in the past dynasties were mainly cleansing， cutting and frying. The processing methods of Citri Reticulatae Pericarpium included in all editions of Chinese Pharmacopoeia were all cleansing and cutting. In modern local processing specifications， there were mainly processing methods such as frying， steaming and carbonizing. The components of Citri Reticulatae Pericarpium were mainly volatile oil and flavonoids. After processing， the total amount of volatile oil generally decreased， and the contents and compositions of volatile oil and flavonoids also changed. It may be the main reason for the difference of efficacy before and after processing. At present， the optimization of processing technology of Citri Reticulatae Pericarpium mainly focuses on steaming， processing with vinegar and baking. Pharmacological research on Citri Reticulatae Pericarpium mainly focuses on reducing blood lipid， relieving cough， relieving asthma and resolving phlegm， and inhibiting pulmonary fibrosis. The current research on the processing methods of Citri Reticulatae Pericarpium is not in-depth enough， the material basis of many processing methods has not been clarified， and the research on the efficacy before and after processing is not in-depth enough. Further research is needed to clarify the material basis and mechanism after the processing of Citri Reticulatae Pericarpium， so as to standardize the processing method and establish specific quality standards.
Keywords：Citri Reticulatae Pericarpium;traditional Chinese medicine;processing specification;chemical composition;volatile oil;flavonoids;pharmacological effect
Abstract：Diabetic neuropathic pain （DNP） is one of the common complications of diabetic peripheral neuropathy， which is difficult to be cured. The clinical manifestations of DNP include reduced pain threshold， neuropathic pain in extremities， and abnormal pain. Studies have demonstrated that nuclear factor-kappa B （NF-κB）， mitogen-activated protein kinase （MAPK）， Toll-like receptor 4 （TLR4）， nuclear factor erythroid 2-related factor 2 （Nrf2）， and phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathways are involved in the treatment of DNP by inhibiting the activation and release of inflammatory cytokines ［e.g.， tumor necrosis factor-alpha （TNF-α） and interleukin （IL）］， regulating inflammation-mediating factors ［e.g.， reactive oxygen （ROS）， inducible nitric oxide synthase （iNOS）， cyclooxygenase （COX-2）， and NF-κB inhibitor （IκB）］， suppressing neurogenic inflammation， destroying the persistent damage of peripheral nerves， and repairing damaged peripheral nerves. The literature of modern medical research has demonstrated that the treatment of DNP with Chinese herbal medicines is associated with the signaling pathways above. They can treat DNP by regulating the expression of key proteins in the signaling pathways to control neurogenic inflammation and repair diabetic peripheral neuropathic damage. By summarizing the available studies， this review aims to provide a reference for the in-depth research on Chinese herbal medicines in the treatment of DNP and give new insights into the protection of diabetic peripheral nerves and the treatment of DNP with Chinese herbal medicines.