Abstract：Tong （dredging） method in traditional Chinese medicine （TCM） emphasizes soothing the stagnated Qi， blood， and body fluid in zang-fu organs， meridians， and collaterals to remove pathogens， reinforce vital Qi， and balance Yin and Yang of the human body. Tong method can be adopted to disperse sweat pore， attack pathogenic Qi， harmonize Yin and Yang， as well as tonify deficiency， and resolve stagnation. It has been proved effective in treating coronary heart disease （CHD）， which falls into the category of "chest impediment and heart pain" in TCM， with the key pathogenesis lying in blood vessel obstruction. Therefore， dredging blood vessels is the primary therapeutic principle for CHD. Specifically， there are four aspects. The first is dispersing and dredging the sweat pore of the heart. If the sweat pore is occluded by pathogenic cold， which makes Yang-qi undissipated， Cinnamomi Ramulus， Piperis Longi Fructus， Alpiniae Officinarum Rhizoma， and Asari Radix et Rhizoma can be prescribed for warming and dredging heart Yang. If the Yang-qi of the heart and chest stagnated in the body， which hinders Qi and blood to nourish the myocardium， resulting in chest pain， Poria and Alismatis Rhizoma can be prescribed. For CHD due to atherosclerosis and inflammation， heat-clearing， toxin-removing， and inflammation-resisting Chinese medicinal herbs such as Coptidis Rhizoma and Rhei Radix et Rhizoma are recommended. The second is attacking and dredging the collaterals of the heart. Salviae Miltiorrhizae Radix et Rhizoma， Chuanxiong Rhizoma， Notoginseng Radix et Rhizoma， etc. can be prescribed for blood stasis， Trichosanthis Fructus， Allii Macrostemonis Bulbus， Pinelliae Rhizoma， etc. for phlegm， and Aquilariae Lignum Resinatum， Euodiae Fructus， etc. for pathogenic cold. Since the chronic disease can affect collaterals， Moschus and Santali Albi Lignum can be added to promote blood circulation and remove the obstruction of collaterals of the heart. The third is harmonizing and dredging the mind. Cinnamomi Ramulus， Coptidis Rhizoma， Cinnamomi Cortex， etc. are selected for restoring the coordination between the heart and the kidney. According to the specific syndrome， the methods of nourishing the mind and calming the nerves through tranquilizing the mind， calming down the mind， and inducing resuscitation can be selected using such Chinese medicines as Ziziphi Spinosae Semen， Polygalae Radix， and Draconis Ossa. The fourth is tonifying and dredging the Qi and blood of the heart. The deficiency syndrome of CHD is divided into Qi deficiency and kidney deficiency. Invigorating Qi and strengthening the heart are the first essentials for the treatment of CHD. In Qi invigoration， Qi and blood must be strengthened simultaneously to strengthen the heart and clear the pulse. Hence， Bazhentang modified by Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos can be chosen. In kidney Qi tonifying， kidney and heart must be strengthened simultaneously， and the methods of tonifying kidney and activating blood can be used. Ginseng Radix et Rhizoma and Astragali Radix are considered as the first choice for tonifying heart Qi， and Epimedii Folium and Morindae Officinalis Radix for tonifying kidney Qi， which are added with Salviae Miltiorrhizae Radix et Rhizoma and Rehmanniae Radix Praeparata to obtain the kidney-tonifying and blood-activating prescription. It is suitable for treating CHD due to kidney deficiency and blood stasis. Simultaneous treatment of heart and kidney is more suitable for middle-aged and elderly patients and chronically ill patients. Tong method can be used in various clinical diseases as well as CHD.
Abstract：Along with increasing degree of population aging globally， senility， good health and long life have become the focus of the world. Guided by Qiluo doctrine， an essence， Qi and spirit theory is proposed as below， essence is the origin of life， Qi is the impetus of life and spirit is the embodiment of life. Based on holistic view of kidney deficiency involving the five internal organs and injuries of the five internal organs definitely affecting the kidney， a mechanism of aging is proposed as below， deficiency of kidney essence is the foundation of aging， deficiency of promordial Qi is the key of aging and physical and spiritual loss is the manifestation of aging. It provides a theoretical guidance for anti-aging study of rejuvenating the elderly and making the strong person stronger. By virtue of the experiences in kidney-tonifying medication accumulated for more than two thousand years， Bazi Bushen capsules has been developed， which has anti-aging efficacy， including tonifying kidney， replenishing essence， coordinating Yin and Yang， supplementing primordial Qi and nourishing body and spirit. Experimental researches have demonstrated that Bazi Bushen capsules can improve overall aging and systemic aging， as well as prevent and treat aging related diseases. Preliminary clinical studies demonstrate that this capsules can enhance athletic ability and improve sexual function， and is expected to become a representative Chinese patent medicine of anti-aging. This paper addresses aging and anti-aging on the basis of Qiluo doctrine， in the hope of helping prevention and treatment of aging related diseases.
Keywords：aging;anti-aging;Qiluo doctrine;Cheng-zhi-tiao-ping;essence， Qi and spirit theory;Bazi Bushen capsules;senile diseases
Abstract：ObjectiveTo observe the effect of Linggui Zhugantang （LG） on the blood-brain barrier （BBB） model of Alzheimer's disease （AD） in vitro and to explore the mechanism of LG in repairing the BBB injury in AD.MethodA total of 50 male SPF rats were randomized into five groups： high-dose （4.8 g·kg-1）， medium-dose （2.4 g·kg-1）， and low-dose （1.2 g·kg-1） LG groups， western medicine （0.5 g·kg-1 donepezil hydrochloride） group， and normal group （normal saline of equivalent volume）. They received （ig） corresponding drugs twice a day for 7 d. Drug-containing serum was respectively collected from the abdominal aorta 1 h after the last administration. The BBB injury of AD in vitro was induced with the cell co-culture method， and 6 groups were designed： normal group， model group， high-， medium-， and low-dose LG groups， and western medicine group. The model group was added with 100 μL amyloid β1-42 （Aβ1-42， final concentration： 5 μmol·L-1）， and high-dose， medium-dose， and low-dose LG groups and the western medicine group were added with corresponding 10% drug-containing serum in addition to the 100 μL Aβ1-42 （final concentration： 5 μmol·L-1）. Cell survival rate was detected by methyl thiazolyl tetrazolium （MTT） assay， expression of BBB-related skeleton proteins （claudin-5， ZO-1， occludin）， matrix metalloproteinase-2 （MMP-2）， and matrix metalloproteinase-9 （MMP-9） by Western blot， and content of inflammatory factors interleukin-1β （IL-1β）， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α） by enzyme-linked immunosorbent assay （ELISA）. BBB Aβ transporter low-density lipoprotein receptor-related protein 1 （LRP-1） and advanced glycation end product receptor （RAGE） at different time points in high-dose， medium-dose， and low-dose LG groups were determined by Real-time PCR and Western blot.ResultCell survival rate of the model group was lower than that of the normal group （P<0.05） and the survival rates of the western medicine group and high-dose LG group was higher than that in the model group （P<0.05）. The skeleton proteins were down-regulated and MMP-2 and MMP-9 were up-regulated in the model group compared with those in the normal group （P<0.05）. The expression of skeleton proteins was higher （P<0.05） and that of MMP-2 and MMP-9 was lower （P<0.05） in the western medicine group and high-dose LG group than in the model group. Compared with the model group， only the medium-dose LG group showed the up-regulation （P<0.05） of claudin-5 （P<0.05） and the decrease （P<0.05） of MMP-2. IL-1β， IL-6， and TNF-α in the model group were up-regulated （P<0.05） compared with those in the normal group， and those inflammatory factors in the western medicine group and high-dose and medium-dose LG groups were lower （P<0.05） than those in the model group. LRP-1 expression was up-regulated and RAGE expression was down-regulated at 3 h compared with those at 0 h （P<0.05）， while the expression of the two became stable at 6， 12， 24， 36 h. At 3 h， LRP-1 expression was down-regulated and RAGE expression was up-regulated in model group compared with those in the normal group at 3 h （P<0.05）. Moreover， the LRP-1 content was higher and RAGE content was lower in the western medicine group and high-dose LG group than in the model group.ConclusionLG can repair the BBB injury in vitro by inhibiting the expression of inflammatory factors and MMP-2， MMP-9， promoting the expression of skeletal proteins， and regulating the balance of transporters.
Abstract：ObjectiveTo investigate the effect of Biejiajian Wan （BJJW） on transforming growth factor-β1 （TGF-β1）-induced epithelial-mesenchymal transition （EMT） of HepG2 cells， and explore its mechanism against EMT of hepatocellular carcinoma cells.MethodHepG2 cells were randomly divided into a blank group， a TGF-β1 model group （10 μg·L-1 TGF-β1）， a low-dose BJJW group （10 μg·L-1 TGF-β1+0.55 g·kg-1 BJJW）， a medium-dose BJJW group （10 μg·L-1 TGF-β1+1.1 g·kg-1 BJJW）， a high-dose BJJW group （10 μg·L-1 TGF-β1+2.2 g·kg-1 BJJW）， and a sorafenib group （10 μg·L-1 TGF-β1+0.03 g·kg-1 sorafenib）. The EMT model was induced by 10 μg·L-1 TGF-β1 inHepG2 cells. After treatment with corresponding medicated serum， cell counting kit -8 （CCK-8） assay was used to detect cell proliferation. Cell migration ability was detected by the Transwell assay and wound healing assay. The protein expression related to EMT and nuclear factor-kappa B （NF-κB） signaling pathway was detected by cell immunofluorescence assay and Western blot.ResultCompared with the blank group 4 days later， the TGF-β1 model group showed fusiform and loose cells with widened gap and antennae reaching out， decreased protein expression of E-cadherin （P<0.05）， and increased protein expression of N-cadherin and vimentin （P<0.05）， which indicated that the EMT model was properly induced in HepG2 cells by TGF-β1 stimulation for 4 days. After 48 hours of treatment with the corresponding medicated serum， each medication group showed inhibited proliferation of HepG2 cells that had undergone EMT， especially the low- and high-dose BJJW groups （P<0.01）， and the medium-dose BJJW group showed increased E-cadherin protein expression （P<0.05） and decreased p-p65， N-cadherin， and vimentin protein expression （P<0.05）， as compared with the TGF-β1 model group. As revealed by the transwell assay and wound healing assay， TGF-β1 enhanced the migration ability of HepG2 cells （P<0.05， P<0.01） compared with the results in the blank group， compared with the TGF-β1 model group， the medication groups showed inhibited migration ability of HepG2 cells （P<0.05， P<0.01）. Compared with the blank group， the TGF-β1 model group promoted the expression of p65 and Snail into the nucleus. Compared with the TGF-β1 model group， the medication groups inhibited the expression of p65 and Snail into the nucleus.ConclusionBJJW may inhibit the EMT， proliferation， and migration of HepG2 cells induced by TGF-β1 by suppressing the NF-κB signaling pathway to exert an anti-hepatocellular carcinoma effect.
Abstract：ObjectiveTo study the effects of Wendantang on the expression of miRNA-219， N-methyl-D-aspartate receptor 2B （NR2B）， disrupted in schizophrenia 1 （DISC1）， and Ca2+/calmodulin-dependent protein kinase Ⅱγ （CaMKⅡγ） in the frontal lobe of rats with schizophrenia.MethodSixty rats were randomly divided into six groups， namely normal group， model group， high-， medium-， and low-dose Wendantang groups， and clozapine group， with 10 rats in each group. Rats in high-， medium-， and low-dose Wendantang groups were intragastric with 40， 20， and 10 g·kg-1 Wendantang， and the ones in clozapine group were intragastric with 0.02 g·kg-1 clozapine， those in normal and model group were intragastric with equal volume of normal saline， once a day. After 21 days of administration， rats in all groups except for the normal group were injected with 0.6 mg·kg-1 dizocilpine maleate （MK-801） into the left abdominal cavity for inducing acute schizophrenia. The stereotypic behavior and ataxia in rats were scored according to SAMS and HOFFMAN criteria. The morphological changes in the prefrontal cortex were observed by hematoxylin-eosin （HE） staining. The protein expression levels of NR2B, DISC1 and CaMKⅡγ in the frontal lobe was detected by Western blot. The mRNA expression levels of miRNA-219 was detected by real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）.ResultCompared with normal group， the model group exhibited significantly increased stereotypic behavior and ataxia scores （P<0.01）， karyopyknosis and karyolysis in most neurons of the prefrontal cortex， and down-regulated NR2B， DISC1， and CaMKⅡγ protein expression （P<0.01） and miRNA-219， NR2B， DISC1， and CaMKⅡγ mRNA expression （P<0.01）. Compared with model group， Wendantang high-， medium-， and low-doses group lowered the scores of stereotypic behavior and ataxia at 50， 60 mmin（P<0.05，P<0.01）. In high- and medium-dose Wendantang groups， the neurons in the prefrontal cortex were densely arranged. The karyopyknosis and karyolysis were alleviated to varying degrees. The NR2B protein expression in the frontal lobe was up-regulated （P<0.01）. In the medium- and low-dose Wendantang groups， the DISC1 protein expression in the frontal lobe was up-regulated （P<0.05，P<0.01）. Wendantang at each dose significantly increased the CaMKⅡγ protein expression （P<0.05） and miRNA-219， NR2B， DISC1， and CaMKⅡγ mRNA expression in the frontal lobe （P<0.05，P<0.01）.ConclusionWendantang improves the scores of stereotypical behavior and ataxia， relieves the karyopyknosis and karyolysis of neurons in the prefrontal cortex， and increases the expression levels of miRNA-219， NR2B， DISC1， and CaMKⅡγ of rats with schizophrenia， so as to alleviate the schizophrenic-like symptoms and schizophrenia.
Keywords：Wendantang;schizophrenia;miRNA-219;N-methyl-D-aspartate receptor 2B （NR2B）;disrupted in schizophrenia 1 （DISC1）
Abstract：ObjectiveTo observe the preventive and control effects of Danggui Niantongtang against adjuvant arthritis differentiated into wind-damp-heat impediment in rats and its influences on the expression of autophagy-related proteins microtubule-associated protein 1 light chain 3 （LC3）， homolog of yeast Atg6 （Beclin1） and p62.MethodThe six-week-old male SD rats were randomly divided into the normal group， wind-damp-heat impediment model group， low-， medium-， and high-dose Danggui Niantongtang （5.67， 11.34， 22.68 g·kg-1） groups， and methotrexate （MTX， 1.35 mg·kg-1） group， with 10 rats in each group. A rat model of adjuvant arthritis was established by subcutaneous injection of inactivated Mycobacterium tuberculosis into the tail root， followed by exposure to the manual climatic box for 16 d for inducing the wind-damp-heat impediment. The drugs were administered intragastrically on the day of immunization for 28 d. The general conditions of rats were observed and the swelling degree of toes and arthritis index （AI） were detected. The pathological changes in the synovial tissues of the knee joints were observed by hematoxylin-eosin （HE） staining. The mRNA expression levels of LC3， Beclin1， and p62 in the synovial tissues were measured by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， followed by the assay of their protein expression by Western blot and immunohistochemistry.ResultCompared with the normal group， the wind-damp-heat impediment model group exhibited significantly increased swelling degree of toes （P<0.01）， increased AI （P<0.01）， proliferated synovial cells （P<0.01）， up-regulated LC3 and Beclin1 protein and mRNA expression （P<0.01）， and down-regulated p62 protein and mRNA expression （P<0.01） after 16， 20， 24， 28-d medication. Compared with the wind-damp-heat impediment model group， each medication group displayed alleviated toe swelling and synovial hyperplasia to different degrees， decreased mRNA and protein expression levels of LC3 and Beclin1 （P<0.01）， and increased p62 mRNA and protein expression （P<0.05，P<0.01）， with the best outcomes observed in the medium-dose Danggui Niantongtang group.ConclusionDanggui Niantongtang effectively relieves adjuvant arthritis due to wind-damp-heat impediment in rats， which may be related to its regulation of the expression of autophagy-related proteins LC3， Beclin1， and p62 and the inhibition of autophagy.
Keywords：Danggui Niantongtang;wind-damp-heat impediment;rheumatoid arthritis;microtubule-associated protein 1 light chain 3 （LC3）;homolog of yeast Atg6 （Beclin1）;p62;autophagy
Abstract：ObjectiveTo study the effects of Chinese herbal compound Youguiwan on angiogenesis of rats with ovarian dysfunction caused by natural aging and its relationship with chemokine interleukin 8 （CXCL8）/CXC chemokine receptor 1/2 （CXCR1/2） signaling pathway， angiopoietin 1 （Ang-1）， and angiopoietin 2 （Ang-2）， so as to explore its mechanism in improving the ovarian function.MethodFifty six female SD rats were randomly divided into the young control group （n=8） and modeling group （n=48， ovarian dysfunction caused by natural aging）. Rats in both the young control and modeling groups were routinely fed， during which the ones in the modeling group underwent exfoliative cytology of vaginal smears for five to seven days. The ones presented with prolonged estrous cycle， followed by continuous estrus and repeated pseudopregnancy revealed by vaginal cytology during four consecutive estrous cycles indicated early aging， and the young rats with keratinocyte proliferation index higher than 50% for 10 consecutive days were classified into the young control group. The successfully modeled rats were randomly divided into the early-aged group， estrogen （65 μg·kg-1·d-1） group， Zuoguiwan （33 g·kg-1·d-1） group， as well as the low-， medium-， and high-dose （1.2， 2.4， 4.8 g·kg-1·d-1） Youguiwan groups. Rats in the young control group and the early-aged group were gavaged with the same volume of normal saline for 30 days. After the experiment， the morphological changes in rat ovary were observed by hematoxylin-eosin （HE） staining. The protein expression levels of chemokines CXCL8， CXCR1， CXCR2， Ang-1， and Ang-2 in rat ovary were detected by Western blotting and immunohistochemistry， and the mRNA expression levels of CXCL8， CXCR1， CXCR2， Ang-1， and Ang-2 by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）.ResultCompared with the young control group， the early-aged group exhibited reduced number of growing follicles， corpus luteum， and blood vessels at all levels， elevated atretic follicles （P<0.01）， up-regulated protein and mRNA expression of CXCL8， CXCR1， and CXCR2 in the ovarian tissue （P<0.01）， and down-regulated Ang-1 and Ang-2 protein and mRNA expression （P<0.05）. Compared with the early-aged group， each medication remarkably increased the number of growing follicles， corpus luteum， and blood vessels （P<0.05）， lowered the number of atretic follicles （P<0.05）， down-regulated the protein and mRNA expression levels of CXCL8， CXCR1， and CXCR2 in the ovarian tissue （P<0.05）， and up-regulated the protein and mRNA expression levels of Ang-1 and Ang-2 （P<0.05）.ConclusionYouguiwan down-regulates the levels of CXCL8， CXCR1， and CXCR2 in rat ovary and up-regulates the levels of Ang-1 and Ang-2 to promote ovarian angiogenesis and improve rat ovarian function.
Abstract：ObjectiveTo explore the effect of Gelsemium elegans combined with Mussaenda pubescens on efflux transporter breast cancer resistance protein （BCRP） and cytochrome P450 3A11 （CYP3A11） and their attenuation mechanism， and to investigate whether the nuclear receptors were involved in such regulation by intervening it with nuclear receptor activators.MethodC57BL/6 mice were divided into the blank group， G. elegans （GE， 0.25 g·kg-1）group， GE + M. pubescens （MP） （0.25 g·kg-1+10 g·kg-1） group， GE + pregnane X receptor （PXR） activator （rifampicin）（GE + Rif，0.25 g·kg-1+50 mg·kg-1） group， GE + MP + Rif （0.25 g·kg-1+10 g·kg-1+50 mg·kg-1） group， GE + constitutive androstane receptor （CAR） activator （1，4-Bis ［2-（3，5-Dichloropyridyloxy）］ benzene， TCPOBOP）（GE + TCP， 0.25 g·kg-1+0.5 mg·kg-1） group， and GE + MP + TCP （0.25 g·kg-1+10 g·kg-1+0.5 mg·kg-1） group. The medication lasted for 14 successive days. One hour after the last administration， the mice were sacrificed by cervical dislocation and the liver tissue was harvested. The left liver tissue was stained with hematoxylin- eosin （HE） for observing the pathological changes. The right liver tissue was used for BCRP and CYP3A11 mRNA and protein expression detection by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot.ResultThe survival rates of mice in the GE + Rif group， GE group， and GE + MP group were 25% （the lowest）， 40%， and 80%， respectively， and no death was observed in the other groups. Compared with the obvious lesions in the liver cells of the GE group， the pathological changes in liver cells of the GE + MP group were alleviated， while those in the GE + Rif group were worsened. Compared with the GE + Rif group， the GE + MP + Rif group exhibited relieved pathological changes in liver cells. Both the GE + TCP group and the GE + MP + TCP group showed mild liver lesions. The comparison with the GE + MP group revealed that the pathological changes in the GE + MP + TCP group were slightly relieved. Compared with the blank group， the expression of BCRP protein and mRNA in GE group were significantly decreased （P<0.05，P<0.01）.The expression of CYP3A11 protein in GE group were significantly decreased （P<0.01）. Compared with the GE group， the GE + MP group displayed remarkably up-regulated BCRP protein and mRNA expression （P<0.05，P<0.01） and CYP3A11 protein expression （P<0.05）， but slightly up-regulated CYP3A11 mRNA expression. Compared with the GE group， the GE + Rif group exhibited down-regulated BCRP protein expression （P < 0.05）. The protein and mRNA expression levels of BCRP were lower in the GE + MP + Rif group than in the GE + MP group （P<0.05，P<0.01）. The PXR activator rifampicin regulated BCRP before and after the combination of G. elegans with M. pubescens. The CYP3A11 protein and mRNA expression levels in the GE + TCP group were higher than those in the GE group （P<0.05，P<0.01）. Compared with the GE + MP group， the GE + MP + TCP group showed up-regulated CYP3A11 protein and mRNA expression （P<0.05，P<0.01）. CAR activator TCPOBOP also had a regulatory effect on CYP3A11 before and after the compatibility of G. elegans with M. pubescens.ConclusionThe attenuated toxin after the combination of G. elegans with M. pubescens is closely related to the efflux transporter BCRP and the drug-metabolizing enzyme CYP3A11.
Keywords：Gelsemium elegans;Mussaenda pubescens;breast cancer resistance protein （BCRP）;cytochrome P450 3A11 （CYP3A11）;nuclear receptor
Abstract：ObjectiveTo screen out the main targets and related signaling pathways of the herbal pair Cremastrae Pseudobulbus-Rhapontici Radix in treating breast cancer based on network pharmacology and verify their action mechanism in in vitro experiments.MethodThe main chemical components and related targets of Cremastrae Pseudobulbus-Rhapontici Radix were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）， and the target genes related to breast cancer from GeneCards. Following the screening of the common targets of Cremastrae Pseudobulbus-Rhapontici Radix and breast cancer using Venn， the Cremastrae Pseudobulbus-Rhapontici Radix-breast cancer network and protein-protein interaction （PPI） network were constructed. The effective targets were then subjected to gene ontology （GO） and Kyoto encyclopedia of genes and genomes （KEGG） pathway enrichment analysis. The resulting outcomes were then verified by cell counting kit （CCK）-8 assay， flow cytometry， and Western blot.ResultThe screening yielded seven effective components and 61 targets of Cremastrae Pseudobulbus-Rhapontici Radix， among which 55 targets were involved in breast cancer. The GO analysis revealed 832 entries， which were mainly enriched in the biological processes. According to KEGG pathway enrichment analysis， 85 signaling pathways were obtained， including tumor suppressor p53， vascular endothelial growth factor （VEGF）， epidermal growth factor receptor （EGFR）， and phosphatidylinositol 3-kinase （PI3K）/protein kinase B（Akt）. It was verified in in vitro experiments that the alcohol extract of Cremastrae Pseudobulbus-Rhapontici Radix inhibited the proliferation of human breast cancer MDA-MB-231 cells and induced their apoptosis. Compared with the blank control group and the dimethyl sulfoxide （DMSO， 0.1% solvent） group， the medication groups exhibited obviously decreased absorbance in MDA-MB-231 cells （P<0.01） and increased apoptosis rate （P<0.01）. The results of Western blot demonstrated that compared with the blank control group and the DMSO group， each medication significantly reduced the phosphorylated （p）-PI3K/PI3K and p-Akt/Akt in cells （P<0.05）.ConclusionThe ethanol extract of Cremastrae Pseudobulbus-Rhapontici Radix effectively inhibits the proliferation of human breast cancer MDA-MB-231 cells and induces their apoptosis， which may be related to the inhibition of the activation of PI3K/Akt signaling pathway.
Abstract：ObjectiveProteoglycan TPG-1 isolated from Trametes robiniophila（Huaier） has proved to have anti-hepatoma activity， and this paper aims to explore the molecular mechanism.MethodHuman hepatoma SK-HEP-1 cells were treated with TPG-1 （0， 0.05， 0.1， 0.25， 0.5， 1 g·L-1）. Then cell survival was detected by methyl thiazolyl tetrazolium （MTT） and apoptosis by flow cytometry. In addition， expression of genes in SK-HEP-1 cells treated with or without TPG-1 was examined by DNA microarray to preliminarily explore the anti-hepatoma molecular mechanism of TPG-1.ResultTPG-1 inhibited the proliferation of SK-HEP-1 cells as compared with the blank group （P<0.01）. After treatment with 1 g·L-1 TPG-1 for 48 h， the apoptosis rate of SK-HEP-1 cells increased （P<0.01）， and TPG-1 promoted the cleavage of cysteinyl aspartate specific proteinase （Caspase）-3 and Caspase-7， the key mediators of apoptosis （P<0.01）. Additionally， TPG-1 （1 g·L-1） suppressed the migration of SK-HEP-1 cells （P<0.05）. A total of 971 differentially expressed genes （DEGs） were identified in SK-HEP-1 cells after treatment with TPG-1， with 486 up-regulated and 485 down-regulated. These DEGs were mainly involved in the Gene Ontology （GO） terms of interleukin-6 （IL-6） biosynthesis， antigen processing and presentation， superoxide dismutase activity， positive regulation of mitogen-activated protein kinase kinase kinase （MAPKKK） cascade， nature killer （NK） cell chemotaxis， and chemokine biosynthesis， and the Kyoto Encyclopedia of Genes and Genomes （KEGG） pathways of nucleotide-binding oligomerization domain （NOD）-like receptor signaling pathway， apoptosis， Toll-like receptor signaling pathway， retinoic acid-inducible gene-Ⅰ （RIG-Ⅰ）-like receptor signaling pathway， T-cell receptor signaling pathway， and chemokine signaling pathway. Western blot results showed that TPG-1 （1 g·L-1） activated mitogen-activated protein kinase （MAPK） signaling pathway in SK-HEP-1 cells （P<0.01）.ConclusionProteoglycan TPG-1 inhibited the proliferation and migration， and induced apoptosis of human hepatoma SK-HEP-1 cells. Up-regulation of MAPK signaling pathway may be responsible for the growth inhibition of human hepatoma SK-HEP-1 cells by TPG-1.
Abstract：ObjectiveTo investigate effect of aqueous extract of Trametes robiniophila （TRM，Huaier） on autophagy of human prostate cancer VCaP cells and Lamin B1 expression， so as to uncover its role in the proliferation of VCaP cells.MethodThe inhibitory effect of 0， 2， 4， 6， 8， 10 g·L-1 TRM aqueous extract on the proliferation of human prostate cancer VCaP cells at different time points were determined by cell counting kit-8 （CCK-8） assay. Acridine orange staining was conducted for analyzing the effect of TRM aqueous extract on the formation of autolysosomes in VCaP cells. After medication， the expression of microtubule-associated protein Ⅰ light chain 3 （LC3）， autophagy-related protein 3 （Atg3）， autophagy-related protein 5 （Atg5）， and autophagy-related protein 7 （Atg7） in VCaP cells were detected by Western blot. The effect of TRM aqueous extract alone and its combination with autophagy inhibitor bafilomycin A1 on the proliferation of VCaP cells were assayed by CCK-8 assay. RNA interference technology was used to explore the role of Lamin B1 in anti-proliferation of VCaP cells by TRM.ResultCompared with the blank group， TRM aqueous extract inhibited the proliferation of human prostate cancer VCaP cells in a time- and concentration-dependent manner （P<0.01）. Acridine orange staining showed that TRM aqueous extract promoted the formation of autolysosomes in VCaP cells. As revealed by Western blotting， TRM aqueous extract up-regulated the expression levels of LC3-Ⅱ， Atg3， Atg5， and Atg7 in contrast to those in the blank group （P<0.05）. All these indicated that TRM aqueous extract induced the autophagy of VCaP cells. In addition， autophagy inhibition impaired the sensitivity of VCaP cells to TRM aqueous extract （P<0.05）. The comparison with the blank group showed that TRM aqueous extract inhibited Lamin B1 protein expression in VCaP cells （P<0.01）， which in turns weakened the sensitivity of VCaP cells to TRM aqueous extract.ConclusionTRM aqueous extract inhibited the proliferation of human prostate cancer VCaP cells possibly by inducing autography and down-regulating Lamin B1 expression. This study has provided a theoretical basis for the clinical application of TRM.
Keywords：aqueous extract of Trametes robiniophila （TRM）;human prostate cancer VCaP cells;autophagy;Lamin B1;cell proliferation
Abstract：ObjectiveTo investigate the effect of Draconis Sanguis petroleum ether fraction （DSPEF） on the proliferation， apoptosis， migration， and autophagy of human gastric cancer HGC-27 and MGC-803 cells， and preliminarily elucidate its molecular mechanism.MethodCell counting kit-8 （CCK-8） assay was used to detect the effect of DSPEF at different concentrations （0， 20， 40， 60， 80 mg·L-1） on the proliferation of HGC-27 and MGC-803 cells after 24， 48， 72 h. Hoechst staining and flow cytometry were used to explore the effects of DSPEF at different concentrations on the apoptosis and apoptosis rate of HGC-27 and MGC-803 cells after 48 h treatment， respectively. The wound healing assay and acridine orange staining were used to investigate the effects of DSPEF on the migration and autophagy of HGC-27 and MGC-803 cells， respectively. Western blot was used to detect the expression levels of signaling pathway-related proteins in HGC-27 and MGC-803 cells treated with DSPEF for 48 h.ResultCompared with the control group， DSPEF（30 mg·L-1） inhibited the proliferation and migration of HGC-27 and MGC-803 cells in a concentration- and time-dependent manner （P<0.05）， and induced the apoptosis （P<0.01） and autophagy of HGC-27 and MGC-803 cells. DSPEF （60 mg·L-1） down-regulated the protein levels of phosphorylated mammalian target of rapamycin （p-mTOR） （P<0.05， P<0.01） and down-regulated phospho-signal transducer and activator of transcription 3 （p-STAT3） in HGC-27 and MGC-803 cells （P<0.01）， suggesting that DSPEF presumedly inhibited the proliferation and migration of human gastric cancer HGC-27 and MGC-803 cells and induced their apoptosis and autophagy by inhibiting the mTOR/STAT3 signaling pathway.ConclusionThe down-regulation of the mTOR/STAT3 signaling pathway may be involved in the anti-gastric cancer effect of DSPEF. This study is expected to provide a reference for the investigation of the anti-tumor effect of Draconis Sanguis.
Keywords：Draconis Sanguis petroleum ether fraction （DSPEF）;human gastric cancer cells;proliferation;apoptosis;migration;autophagy
Abstract：Malignant tumor is a serious threat to human life and health. The prevalence and mortality of malignancies in China are increasing year by year. Conquering cancer has become a difficult problem for human beings. Chemical drug therapy combined with molecular targeted therapy is a general and preferred anti-tumor clinical scheme， but the side effects and the drug resistance of cancer cells often hinder the efficacy. Therefore， it is of great significance to study the mechanism of drug resistance and the methods to reverse drug resistance. Chinese medicine has the characteristics of complex components， multiple targets， low toxicity， etc. A large number of experimental studies have demonstrated that the effective components or extracts of Chinese medicine can inhibit the proliferation， migration， and invasion of cancer cells， and induce apoptosis， autophagy， differentiation， and senescence. In clinical practice， Chinese medicine has been applied to the protection against ttumor， adjuvant treatment， and later consolidation. The research on Chinese medicine is expected to promote drug resistance reversal and cancer therapy. Studies have shown that the combination of Chinese medicine and chemotherapy can reverse drug resistance and increase efficacy， which has become the mainstream trend of cancer treatment. This study reviewed the mechanisms of the drug resistance of cancer cells induced by self-protective autophagy， gene mutation， high expression of enzymes， abnormal signaling pathways， and abnormal expression of RNA and protein， and summarized how compounds isolated from Chinese medicine， single drug and its extract， and classic anti-cancer prescription reversed the drug resistance to lay a solid foundation for the further investigation of the anti-tumor effect of Chinese medicine.
Keywords：cancer;reverse drug resistance;Chinese medicine;mechanisms
Abstract：ObjectiveOn the basis of determining the protective effect of berberine （BBR） on cerebral ischemia， crucial transcription factors （TFs） of BBR against cerebral ischemia was identified by using transcriptome and proteome sequencing.MethodThe model of middle cerebral artery occlusion （MCAO） was established by thread embolization. The sham operation group， model group， low-dose group of BBR （dose of 37.5 mg·kg-1·d-1） and high-dose group of BBR （75 mg·kg-1·d-1） were set up. The rats were killed after continuous intragastric administration for 7 days. The pharmacodynamics was evaluated by Longa score and cerebral infarction rate， and the expressions of inflammatory cytokines， such as interleukin （IL）-1β， tumor necrosis factor （TNF）-α and monocyte chemotactic protein-1 （MCP-1） were measured by enzyme-linked immunosorbent assay （ELISA）. Then， RNA-Seq technique was used to detect the differentially expressed genes （DEGs） before and after BBR intervention， and DAVID 6.8 was used for enrichment analysis of DEGs. CatTFREs technique was used to detect differential TFs before and after BBR intervention， and DAVID 6.8 and STRING 11.0 were used for enrichment analysis and TFs association analysis. Finally， by integrating the activity of TFs and the changes of downstream target genes， crucial TFs were identified and the related regulatory network was constructed by Cytoscape 3.7.1.ResultCompared with the sham operation group， the neurological impairment was significant in the model group （P<0.01）， and compared with the model group， the low and high dose BBR groups could significantly reduce the neurological function damage （P<0.01） and decrease the rate of cerebral infarction （P<0.01）. Transcriptome data analysis showed that BBR was involved in the recovery process after cerebral ischemia mainly by affecting cell adhesion， brain development， neuron migration， calcium signaling pathway， cyclic adenosine monophosphate （cAMP） signaling pathway， inflammatory response and other related functions and signaling pathways. Proteomic data analysis showed that the differentially expressed TFs after BBR intervention interfered with cerebral ischemia mainly by regulating cell differentiation， immune system process， cell proliferation and other biological processes. In addition， integration analysis of TFs and DEGs revealed that transcription factor CP2-like 1 （TFCP2L1）， nuclear factor erythroid-2 like 1 （NFE2L1）， neurogenic differentiation protein 6 （NeuroD6） and POU domain， class 2， transcription factor 1 （POU2F1） were crucial TFs against cerebral ischemia-reperfusion injury mediated by BBR.ConclusionBBR has obvious protective effect on cerebral ischemia-reperfusion injury and its crucial TFs include TFCP2L1， NFE2L1， NeuroD6 and POU2F1.
Keywords：berberine;cerebral ischemia;RNA-Seq;catTFREs;transcription factors;inflammatory factors;middle cerebral artery occlusion （MCAO） model
Abstract：ObjectiveTo explore the regulatory effect of Quyu Huatan Tongmai prescription on intestinal mircoflora of hyperlipidemia golden hamster and scientific evidence for the compatibility.MethodSyrian golden hamsters were randomized into normal， model， prescription， stasis-dispelling （Quyu）， phlegm-dissolving （Huatan）， and detoxification （Jiedu） groups， with 8 in each group. Hyperlipidemia in golden hamsters was induced by high-fat diet （4 weeks）. Then hamsters in the Quyu group （1.11 g·kg-1）， Huatan group （0.39 g·kg-1）， Jiedu group （0.07 g·kg-1）， and prescription group （1.42 g·kg-1） were given （ig） corresponding drugs and those in the normal group and the model group received （ig） distilled water of equivalent volume， once a day for 6 weeks. Serum lipids were determined， and hematoxylin-eosin （HE） staining was used to observe the pathological morphology of the liver. Feces were collected for 16S rRNA gene high-throughput sequencing of intestinal flora.ResultCompared with normal group， the model group demonstrated increase in body weight （P<0.05， P<0.01） and blood lipids （P<0.01）， decrease in intestinal flora diversity （P<0.05， P<0.01）， and variation of the relative abundance of intestinal flora at phylum， family， and genus levels （P<0.05， P<0.01）. Compared with the model group， Quyu Huatan Tongmai prescription controlled the body weight change， reduced the serum triglyceride （TG）， total cholesterol （TC）， and low density lipoprotein cholesterol/high density lipoprotein cholesterol ratio （LDL-C/HDL-C） （P<0.05， P<0.01）， improved the structure of intestinal flora， decreased the ratio of Firmicutes to Bacteroides （P<0.01）， raised the abundance of Bacteroidaceae， Porphyromonadaceae， Rikenellaceae， and Pasteurella （P<0.05， P<0.01）， and lowered the relative abundance of Coriobacterium （P<0.05） in hyperlipidemia golden hamsters. All the split prescriptions improved blood lipids and intestinal flora of the hamsters and particularly， the lipids-lowering effect of the Jiedu group and the regulation of flora in the Huatan group were closer to those of the prescription group.ConclusionQuyu Huatan Tongmai prescription and the split prescriptions all alleviated the hyperlipidemia of golden hamsters to different degrees possibly by regulating intestinal flora structure and improving intestinal microecology. The effect of the prescription group was most significant， and coming in second was the Huatan group. This study also provides scientific evidence for the effect of Quyu Huatan Tongmai prescription.
Abstract：ObjectiveTo study the effect of Huazhuo Jiedu Huoxue Tongluo （HJHT） prescription on the intestinal flora in rats with cerebral ischemia-reperfusion injury， and to explore the mechanism of Chinese medicinal prescription regulating intestinal flora to restore the balance of brain-gut axis.MethodFifty male SPF SD rats were randomly assigned into sham group， model group， high-dose HJHT group （25.0 g·kg-1）， medium-dose HJHT group （12.5 g·kg-1）， and low-dose HJHT group （6.25 g·kg-1）， with 10 rats in each group. The rat model of permanent middle cerebral artery infarction was established according to Longa method and previous research experience， and reperfusion was performed 2 h after ischemia. The recovery of neurological function deficit and the percentage of cerebral infarction area were detected 72 h after administration. Real-time PCR was performed to detect the mRNA levels of Occludin and zonula occludens-1 （ZO-1） in rat colon. Hematoxylin-eosin （HE） staining was conducted to reveal the intestinal damage. The feces of 6 rats in each group were collected for 16S rRNA sequencing. The expression of Treg and Th17 in intestinal tissue， peripheral blood， and brain tissue were detected.ResultCompared with the sham group， the model group showed obvious neurological deficit （P<0.05） and large cerebral infarction area （P<0.05）. High-dose and medium-doses HJHT alleviated the symptoms of neurological impairment （P<0.05） and reduce the cerebral infarction area （P<0.05） compared with the model group. Compared with the sham group， the model group showed destroyed structure of colonic mucosa and incomplete epithelial cells and goblet cells， while high-dose and medium-doses HJHT alleviated such changes. The mRNA levels of Occludin and ZO-1 in the model group were lower than those in the sham group （P<0.05），and the high-dose HJHT groups were higher than the model group （P<0.05）. The intestinal flora structure was significantly different between the model group and the sham group while similar between the high-dose HJHT group and sham group. Compared with the sham group， the model group showed down-regulated expression of Treg and up-regulated expression of Th17 in the intestinal tissue， peripheral blood， and brain tissue， and high-dose and medium-dose HJHT alleviated the changes in the expression of Treg and Th17 in the model group （P<0.05）.ConclusionHuazhuo Jiedu Huoxue Tongluo prescription may improve the permeability of intestinal wall by adjusting the abundance and diversity of intestinal microorganisms to reduce the migration of intestinal Th17 cells toward the ischemic lateral brain tissue， mitigate the inflammatory response， and thus alleviate the cerebral ischemia-reperfusion injury in rats.
Abstract：ObjectiveTo explore the effects of different treatment methods of "soothing liver， invigorating spleen， soothing liver and invigorating spleen， soothing liver first and then soothing liver and invigorating spleen， as well as invigorating spleen first and then soothing liver and invigorating spleen" on liver depression combined with liver injury in rats and their action mechanisms.MethodA six-week rat model of liver depression combined with liver injury was established by restraint stress and subcutaneous injection of carbon tetrachloride （CCl4， 5.89 g·kg-1， once every three days）. At the same time， the drugs were given by gavage. Forty-eight male SD rats of clean grade were randomly divided into eight groups， namely the normal group， model group， bicyclol （0.2 g·kg-1） group， Sinisan （4.32 g·kg-1） group， Liu Junzitang （9.26 g·kg-1） group， Chaishao Liu Junzitang A （Chai A， soothing liver and invigorating spleen，13.57 g·kg-1） group， Chaishao Liu Junzitang B （Chai B， soothing liver first and then soothing liver and invigorating spleen， 13.57 g·kg-1） group， and Chaishao Liu Junzitang C （Chai C， invigorating spleen first and then soothing liver and invigorating spleen， 13.57 g·kg-1） group， with six rats in each group. The pathological changes in liver and colon tissues of each group were observed under light microscope and electron microscope. The serum biochemical indexes of the liver were detected using an automatic biochemical analyzer. The relative mRNA expression levels of Takeda G protein-coupled receptor 5 （TGR5） and intestinal mucosal zona occluden-1 （ZO-1）， Occludin， and Claudin-1 in the liver and colon were detected by reverse-transcription polymerase chain reaction （RT-PCR）. The positive expression rate of proliferating cell nuclear antigen （PCNA） in the colon was detected by immunohistochemistry.ResultCompared with normal group， the model group exhibited significantly elevated serum alkaline phosphatase （ALP）， alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， total bilirubin （TBIL）， and direct bilirubin （DBIL） （P<0.01）， lowered TGR5 mRNA expression in liver tissue， up-regulated TGR5 mRNA expression in the colon tissue （P<0.05，P<0.01）， and down-regulated ZO-1， Occludin， and tight junction protein-1 (Claudin-1) mRNA expression and PCNA in the colon tissue （P<0.01）. Compared with the model group， bicyclol and Chai C remarkably decreased the levels of serum ALP， ALT， AST， TBIL， and DBIL （P<0.05，P<0.01）， while Liu Junzitang， Chai A， Chai B， and Chai C significantly up-regulated the TGR5 mRNA expression in the liver and down-regulated its expression in the colon （P<0.01）. Bicyclol， Chai A， Chai B， and Chai C enhanced the ZO-1 and Claudin-1 mRNA expression in the colon （P<0.05，P<0.01）. Bicyclol， Sinisan， and Chai C increased PCNA expression （P<0.01）. The comparison with the Chai C group showed that the TGR5 mRNA expression in the liver and ZO-1 mRNA expression in the colon of the bicyclol and Sinisan groups were lower， whereas the TGR5 mRNA expression in the colon was higher （P<0.01）. However， the PCNA expression in the colon of the Liu Junzitang and Chai B groups declined significantly （P<0.05）.ConclusionIn the presence of liver injury， invigorating spleen first helps to relieve the liver injury， and the efficacy of "spleen-invigorating" therapy in increasing the intestinal mucosal tight junction proteins and improving the gastrointestinal function is related to its activation of TGR5 to improve the intestinal mucosal barrier function， promote the renewal of intestinal stem cells， and drive the regeneration after injury.
Keywords：liver disease transmitting into spleen;soothing liver and invigorating spleen;Takeda G protein-coupled receptor 5 （TGR5）;zona occluden-1 (ZO-1);Occludin;tight junction protein-1 (Claudin-1);colonic proliferating cell nuclear antigen （PCNA）
Abstract：ObjectiveTo explore the mechanism of Shenxiong glucose injection （SGI） in inhibiting hydrogen peroxide （H2O2）-induced oxidative damage in H9c2 cells by tandem mass tags （TMT）-labeled quantitative proteomics.MethodH9c2 cells cultured in vitro were exposed to H2O2 for inducing oxidative damage. The cell viability was measured by cell proliferation and cytotoxicity assay （MTS）， followed by peptide fractionation by high performance liquid chromatography （HPLC） and protein expression detection in H9c2 cells by ultrahigh performance liquid chromatography-mass spectrometry. MaxQuant （v22.214.171.124） was utilized for data retrieval， and the high-resolution mass spectrometry was conducted to screen out differentially expressed proteins， which were then subjected to gene ontology （GO） and Kyoto encyclopedia of genes and genomes （KEGG） enrichment analysis. The protein expression levels of perilipin 2 （Plin2） and tropomyosin 1 （Tpm1） in cells were measured by Western blot.ResultThe spectral analysis yielded 48 608 specific peptide fragments and 5 903 quantifiable proteins. Compared with the model group，the SGI group exhibited 82 differentially expressed proteins，of which 22 were up-regulated and 60 were down-regulated. GO analysis results showed that the differentially expressed proteins were mainly involved in biological processes such as programmed cell death regulation，regulation of cell proliferation，cardiovascular system development， and cell migration. As revealed by KEGG analysis， these proteins were mainly related to peroxisome proliferator-activated receptor （PPAR），focal adhesion，phosphatidylinositol 3-kinase/protein kinase B（PI3K/Akt），and Ras-related protein 1 （Rap1） pathways. Western blot results demonstrated that compared with the model group，SGI significantly increased the Plin2 protein expression and decreased the Tpm1 protein expression （P<0.01），consistent with the proteomics results.ConclusionSGI may inhibit cell apoptosis and antagonize H2O2-induced cell oxidative damage by regulating PPAR，focal adhesion，PI3K/Akt and Rap1 pathways，which should be further verified by subsequent experiments.
Abstract：ObjectiveTo retrospectively analyze the clinical data of 52 patients with coronavirus disease-2019 （COVID-19） and explore the clinical efficacy of modified Sanxiaoyin on mild/moderate COVID-19 patients.MethodThe propensity score matching method was used to collect the clinical data of mild or moderate COVID-19 patients enrolled in the designated hospital of the Second Hospital of Jingzhou from December 2019 to May 2020. A total of 26 eligible patients who were treated with modified Sanxiaoyin were included in the observation group， and the 26 patients treated with conventional method were the regarded as the control. The disappearance of clinical symptoms， disappearance time of main symptoms， efficacy on traditional Chinese medicine （TCM） symptoms， hospitalization duration， laboratory test indicators， and CT imaging changes in the two groups were compared.ResultThe general data in the two groups were insignificantly different and thus they were comparable. After 7 days of treatment， the disappearance rate of fever， cough， fatigue， dry throat， anorexia， poor mental state， and poor sleep quality in the observation group was higher than that in the control group （P<0.05）， and the difference in the disappearance rate of expectoration and chest distress was insignificant. For the cases with the disappearance of symptoms， the main symptoms （fever， cough， fatigue， dry throat， anorexia， chest distress） disappeared earlier in the observation group than in the control group （P<0.01）. After 7 days of treatment， the scores of the TCM symptom scale of both groups decreased （P<0.01）， and the decrease of the observation group was larger that of the control group （P<0.01）. All patients in the two groups were cured and discharged. The average hospitalization duration in the observation group ［（12.79±2.68） d］ was shorter than that in the control group ［（15.27±3.11） d］ （P<0.01）. The effective rate in the observation group （92.31%， 24/26） was higher than that in the control group （76.92%， 20/26） . After 7 days of treatment， the lymphocyte （LYM） count increased （P<0.05）， and white blood cell （WBC） count and neutrophil （NEUT） count decreased insignificantly in the two groups. Moreover， levels of C-reactive protein （CRP）， erythrocyte sedimentation rate （ESR）， and procalcitonin （PCT） reduced in the two groups after treatment （P<0.01） and the reduction in the observation group was larger than that in the control group （P<0.01）. Through 7 days of treatment， the total effective rate on pulmonary shadow in the observation group （90.00%， 18/20） was higher than that in the control group （77.27%， 17/22） （P>0.05） and the improvement of lung shadow in the observation group was better than that in the control group （P<0.01）.ConclusionModified Sanxiaoyin can significantly alleviate fever， cough， fatigue， anorexia， chest distress， poor sleep quality， and other symptoms of patients with mild or moderate COVID-19， improve biochemical indicators， and promote the recovery of lung function. This paper provides clinical evidence for the application of modified Sanxiaoyin in the treatment of mild or moderate COVID-19.
Keywords：modified Sanxiaoyin;coronavirus disease-2019 （COVID-19）;clinical research
Abstract：ObjectiveTo investigate the antidiarrheal effect and mechanism of Zingiberis Rhizoma Recens on diarrhea mice， and to provide research basis for the inhibition of intestinal peristalsis by Zingiberis Rhizoma Recens and its application in the treatment of gastrointestinal diseases.MethodThe diarrhea model of mice was established by Sennae Folium. The control group， model group， Zingiberis Rhizoma Recens low-， medium-， high-dose groups （0.1， 0.32， 1.0 g·kg-1） and loperamide group （1.6 g·kg-1） were set. The intervention effect of Zingiberis Rhizoma Recens with different doses on diarrhea mice was detected by diarrhea score， incidence rate of loose stools （LSIR）， grade of average loose stools （ALSG）， diarrhea index （DI）， intestinal propulsion rate and intestinal pathological section. The serum metabonomics of mice was analyzed by ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry （UPLC-QE-Orbitrap-MS）， principal component analysis （PCA） and orthogonal partial least squares discriminant analysis （OPLS-DA）. The conditions were as follows：mobile phase of 0.1% formic acid aqueous solution （A）-0.1% formic acid acetonitrile solution （B） for gradient elution （0-3.5 min， 5%-15%B； 3.5-6 min， 15%-30%B； 6-6.5 min， 30%B； 6.5-12 min， 30%-70%B； 12-12.5 min， 70%B； 12.5-18 min， 70%-100%B）， flow rate of 0.4 mL·min-1， injection volume of 5 µL， electrospray ionization （ESI）， positive and negative ion detection modes， acquisition range of m/z 100-1 500.ResultCompared with the model group， Zingiberis Rhizoma Recens high-dose group could obviously reduce the diarrhea score， LSIR， ALSG， DI and intestinal propulsion rate （P<0.05， P<0.01）， and improve the intestinal mucosal injury. There were 40 main differential metabolites among the control group， model group and Zingiberis Rhizoma Recens high-dose group， including glucose 1-phosphate， xanthine， xanthosine and so on. The metabolic pathways mainly included starch and sucrose metabolism， amino sugar and nucleotide sugar metabolism， fructose and mannose metabolism， tryptophan metabolism， and galactose metabolism.ConclusionZingiberis Rhizoma Recens can inhibit intestinal peristalsis in diarrhea mice and exert antidiarrhoea effect， the mechanism of which may be related to the regulation of carbohydrate and amino acid metabolism.
Keywords：Zingiberis Rhizoma Recens;diarrhea;metabonomics;differential metabolites;metabolic pathways;mechanism;ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry （UPLC-QE-Orbitrap-MS）
Abstract：ObjectiveTo study the chemical constituents of the seeds of Sophora tonkinensis.MethodThe chemical constituents were isolated and purified by chromatography with MCI resin， silica gel， Sephadex LH-20， and semi-preparative high performance liquid chromatography. Their structures were identified by physicochemical properties， spectral data as well as relevant references. Meanwhile， the antibacterial activities against Helicobacter pylori of these compounds were screened by agar dilution method.ResultA total of 22 compounds were isolated from the methanol extract of the seeds of S. tonkinensis， and characterized as 4′，7-dihydroxy-6-methoxy isoflavone （1）， daidzein （2）， wighteone （3）， dalparvone （4）， 5，7-dihydroxy-4′-methoxyisoflavone （5）， prunetin （6）， formononetin （7）， genistein （8）， 5-methoxydaidzein （9）， ononin （10）， 7，4′-dihydroxyflavone （11）， liquiritigenin （12）， bayin （13）， 2，4-dihydroxybenzoate （14）， methyparaben （15）， 4-hydroxyacetophenone （16）， p-anisaldehyde （17）， methyl indole-3-carboxylate （18）， 4-［β-D-apiofuranoyl-（1→6）-O-β-D-glucopyranosyloxy］ phenylacetonitrile （19）， （-）-methyl dihydrophaseate （20）， methyl canavaliol ester （21）， vomifoliol 3′-O-β-D-apiofuranosyl-（1→6）-β-D-glucopyranoside （22）.ConclusionCompounds 1， 5， 6， 9 and 16 are isolated from S. tonkinensis for the first time， compounds 4， 14， 17-22 are isolated from the genus of Sophora for the first time. In addition， compounds 10 and 13 display moderate antibacterial activities against H. pylori.
Keywords：Sophora;seeds of S. tonkinensis;chemical constituents;flavonoids;isoflavonoids;terpenoids;Helicobacter pylori
Abstract：ObjectiveTo explore the role of transient receptor potential vanilloid 1 （TRPV1） channel in reducing cardiomyocyte toxicity of Aconiti Kusnezoffii Radix processed with Chebulae Fructus.MethodH9c2 cardiomyocytes cultured in vitro were used as a model to assess cell viability by methyl thiazolyl tetrazolium （MTT） assay， the expression of TRPV1 mRNA was detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， and the leakage rate of lactate dehydrogenase （LDH）， the changes of nucleus， reactive oxygen species （ROS）， mitochondrial membrane potential and Ca2+ contents were detected by enzyme linked immunosorbent assay （ELISA）.ResultCompared with the blank group， when the concentration was ≥0.5 g·L-1， the cell viability was significantly decreased （P<0.01）， the leakage rate of LDH， the release of ROS and Ca2+ were increased， the mitochondrial membrane potential was decreased， and the nucleus was pyknosis or even broken in raw Aconiti Kusnezoffii Radix and Aconiti Kusnezoffii Radix processed with Chebulae Fructus groups. When the concentration was ≥0.5 g·L-1， compared with the same mass concentration of raw Aconiti Kusnezoffii Radix group， the cell viability increased significantly （P<0.01）， the leakage rate of LDH， the release of ROS and Ca2+ decreased， the mitochondrial membrane potential increased， and the nuclear morphology improved in Aconiti Kusnezoffii Radix processed with Chebulae Fructus group. Application of the same mass concentration of raw Aconiti Kusnezoffii Radix to H9c2 cardiomyocytes pretreated with the TRPV1 inhibitor BCTC significantly increased cell viability， decreased leakage rate of LDH， ROS and Ca2+ release， increased mitochondrial membrane potential and improved nuclear pyknosis compared with untreated H9c2 cardiomyocytes. Application of the same mass concentration of Aconiti Kusnezoffii Radix processed with Chebulae Fructus to H9c2 cardiomyocytes pretreated with BCTC decreased cell viability， increased LDH leakage rate， ROS and Ca2+ release， reduced mitochondrial membrane potential compared with untreated H9c2 cardiomyocytes. Real-time PCR results showed that both raw Aconiti Kusnezoffii Radix and Chebulae Fructus decoction could increase the expression of TRPV1 mRNA in cardiomyocytes in a concentration dependent manner.ConclusionRaw Aconiti Kusnezoffii Radix can induce cardiomyocyte apoptosis and cardiotoxicity by activating TRPV1 channel， while Aconiti Kusnezoffii Radix processed with Chebulae Fructus can attenuate the toxicity through TRPV1 channel， which may be related to the synergistic effect of acid components in Chebulae Fructus and alkaloids in Aconiti Kusnezoffii Radix on TRPV1 channel.
Abstract：ObjectiveTo establish a simple， fast and accurate method for locating the volatile oil in Angelicae Sinensis Radix based on frozen section and fluorescence imaging technology， and to reveal the distribution and accumulation of volatile oil in the roots of this herbal medicine.MethodAngelicae Sinensis Radix was used as the research material， the best frozen section conditions for the research material were established by comparing the effects of different cryoprotectants on the quality of frozen sections of Angelicae Sinensis Radix. The suitability of Sudan Ⅲ chemical staining and fluorescence localization for positioning the volatile oil were compared according to the loss of volatile oil and the complexity of operation process.ResultA new method for evaluating the quality of frozen sections of Angelicae Sinensis Radix was established. According to the evaluation equation， it was found that the highest score was obtained when the head， body and tail positions of Angelicae Sinensis Radix were treated with 20% glycerol， 15% glycerol and 20% sucrose， respectively. There was yellowish-brown oily substance in the oil chambers of phelloderm and secondary phloem， and oil canal of the secondary xylem of Angelicae Sinensis Radix， which could be stained orange red or orange yellow by Sudan Ⅲ， and there was green spontaneous fluorescence in the same part under the fluorescence microscope.ConclusionThe relatively complete section of Angelicae Sinensis Radix can be obtained after being treated with cryoprotectant. The volatile oil exists in the oil chambers of phelloderm and secondary phloem， and oil canal of the secondary xylem of Angelicae Sinensis Radix. This study can provide reference for observation of the accumulation sites of volatile oil in other plants.
Abstract：ObjectiveTo collect and screen records concerning the spleen and stomach diseases and syndromes in ancient books of traditional Chinese medicine （TCM） using an automated framework and to systematically explore the concept evolution of spleen and stomach diseases and syndromes using the visualization method.MethodA total of 1 224 kinds of ancient book data in the Ancient Books of Traditional Chinese Medicine Database （V2.1） were analyzed using the automated testing tool Selenium WebDriver with the Python 3.8 programming language and the etree library of Lxml for automatic collection and statistics of the "book title" "author" "classification" "dynasty" "completion time"， and "informative abstract". After being checked and collated， the collected data were visually analyzed with Tableau （V2020.1.3） for figuring out the concept evolution of spleen and stomach diseases and syndromes in the past dynasties from the perspectives of symptoms and signs， etiology and pathogenesis， principle-method-recipe-medicinal， and prognosis.ResultA total of 7 203 clauses were automatically collected from 989 ancient books. It was found that in the pre-Qin period， there were few ancient books related to the spleen and stomach diseases and syndromes， and the understandings were confined to the superficial symptoms or signs and the basic etiology and pathogenesis. From the Han to Sui and Tang dynasties， the related concepts gradually increased and the descriptions about the manifestations are more detailed than those in previous dynasties. The etiology， diagnosis， and treatment system of the spleen and stomach diseases and syndromes were further perfected. In the Song， Jin， and Yuan dynasties， such concepts as independent signs，symptoms， as well as nature and location of spleen and stomach diseases and syndromes were enriched. In the Ming and Qing dynasties， a TCM syndrome differentiation and treatment system for spleen and stomach diseases and syndromes was formed， and the related concepts were gradually simplified and unified.ConclusionThe concepts of spleen and stomach diseases and syndromes have undergone an evolution from simplicity to complexity and then back to simplicity. There are numerous ancient books discussing the concepts of spleen and stomach diseases and syndromes， exhibiting a fluctuating yet rising trend with time. The automated framework enables the construction of a lightweight database of spleen and stomach diseases and syndromes. Based on data visualization， the concept evolution of the spleen and stomach diseases and syndromes from ancient times to the present has been efficiently uncovered， which is conducive to tracing the origin and development of spleen and stomach diseases and syndromes in TCM. This has provided reference for related research of spleen and stomach diseases in modern Chinese medicine.
Keywords：concepts of spleen and stomach diseases and syndromes in traditional Chinese medicine （TCM）;automated framework;visualization analysis;Selenium;Tableau
Abstract：ObjectiveTo predict the therapeutic targets and related signaling pathways of orcinol glucoside （OG） in the treatment of osteoporosis by network pharmacology， and further clarify its mechanisms based on molecular docking and in vitro cell model.MethodThe pharmacological targets of OG were obtained from Similarity ensemble approach （SEA） and SwissTargetPrediction， and the targets related to osteoporosis from DisGeNET and GeneCards. The cross-analysis was conducted to screen the common targets between OG and osteoporosis. STRING was used to construct the protein-protein interaction （PPI） network， followed by topology analysis using CytoNCA plug-in of Cytoscape 3.7.2 to screen out the core targets. The obtained common targets were subjected to gene ontology （GO） and Kyoto encyclopedia of genes and genomes （KEGG） analysis by g：Profiler. AutoDock Vina was utilized for molecular docking， and the in vitro cell experiments were then carried out for verifying the mechanism of OG in treating osteoporosis.ResultA total of 73 targets related to OG and osteoporosis were harvested，among which 14 were proved to be key targets by topological analysis. GO and KEGG functional enrichment analysis yielded 259 cell biological processes， mainly involving organonitrogen compound metabolic process， cell population proliferation， protein metabolic process， regulation of response to stress， and response to chemicals. Its mechanism of action might be related to advanced glycation end-product （AGE）-AGE receptor （RAGE） signaling pathway， interleukin-17 （IL-17） signaling pathway， and phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway. Molecular docking indicated that the binding energies of OG to Cyclin D1 （CCND1） and cyclin-dependent kinase 4 （CDK4） were the lowest and similar. The results of flow cytometry showed that compared with the normal group， OG group exhibited decreased proportion of cells in G0/G1 phase （P<0.01） and decreased proportion of cells in S phase （P<0.01）. As demonstrated by Western blot， compared with the normal group， OG up-regulated the protein expression levels of Cyclin D1 and CDK4 （P<0.05， P<0.01）.ConclusionOG alleviates osteoporosis via multiple targets and multiple pathways. It may exert the therapeutic effects by increasing Cyclin D1 and CDK4 protein expression to change cell cycle and promote cell proliferation.
Abstract：ObjectiveTo preliminarily predict the active components， action targets， and signaling pathways of Arnebia euchroma in the treatment of melanoma based on network pharmacology and molecular docking， and to verify its possible mechanism of action in in vitro experiments.MethodThe active components and related targets of A. euchroma were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）SwissTargetPrediction and literature， and the targets related to melanoma from the GeneCards， Online Mendelian Inheritance in Man （OMIM）， and Comparative Toxicogenomics Database （CTD）. Following the construction of the protein-protein interaction （PPI） network of active components and related targets of A. euchroma and melanoma-related targets using STRING， Cytoscape 3.8.2 was used for screening and analyzing the nodes in the network of A. euchroma against melanoma. The intersections were subjected to gene ontology （GO） and Kyoto encyclopedia of genes and genomes （KEGG） enrichment analysis using DAVID 6.8. Acetyl alkannin， the active component in A. euchroma， was docked to the target by AutoDock Vina 1.1.2. The in vitro experiments were then carried out to verify the anti-melanoma effect of A. euchroma.ResultA total of 271 common targets of A. euchroma and melanoma were harvested， among which 23 were key targets， including matrix metalloproteinase-9 （MMP-9） and Janus kinase 2 （JAK2）. As revealed by KEGG enrichment analysis， A. euchroma mainly acted on Janus kinase/signal transduction and activator of transcription （JAK/STAT）， tyrosine kinase receptor （ErbB）， and vascular endothelial growth factor （VEGF） signaling pathways to resist melanoma. According to molecular docking， acetyl alkannin exhibited a good docking activity with JAK2， STAT3， VEGF， MMP-9， and E-cadherin receptors. The results of Western blot and Real-time quantitative polymerase chain reaction （Real-time PCR） showed that acetyl alkannin at different doses inhibited the protein and gene expression of JAK2， STAT3， VEGF， MMP-9， and E-cadherin in A375 cells （P<0.05）.ConclusionA. euchroma alleviates melanoma via multiple targets and multiple pathways， and it may exert the therapeutic effects by affecting the expression of such key target proteins as JAK2， STAT3， VEGF， MMP-9， and E-cadherin and inhibiting the invasion and metastasis of melanoma cells. This study has provided an experimental basis for the treatment of tumor with A. euchroma.
Abstract：In this paper， the key technical problems in the research and development of famous classical formulas are analyzed. Firstly， the puzzled problem for a long-time， which is conversion relationship from medicinal metrology of Han dynasty （HD） to that of modern （gram，g）， is comprehensively expounded that one Liang （两） of HD=3 g is more appropriate. Secondly， the model and principles of quality consistency evaluation are given for the transformation from the quality of authoritative basic sample prepared by casserole （ABS-C） to the quality consistency in Laboratory process， pilot-scale process and industrial production. The consistency evaluation model is ξABS-X=K1（Q1ABS-X/Q1ABS-C）+K2（Q2ABS-X/Q2ABS-C）+……+Ki（QiABS-X/QiABS-C）=∑Ki（QiABS-X/QiABS-C）（i=1，2，3……n）. In the formula， ABS-X means laboratory reference sample ABS-C （ABS-L）， pilot-scale ABS-C （ABS-mP） or industrial production ABS-C （ABS-P）， ξABS-X means the quality consistency rate or similarity degree of ABS-L， ABS-mP and ABS-P processes with ABS-C， Ki means the weight of each quality evaluation index （i）， QiABS-X is the data of i in ABS-L， ABS-mP， ABS-P samples， and QiABS-C is the data （or mean） of i in ABS-C sample. Thirdly， in order to control the quality of the herbal medicines whose active ingredients were unknown， their chemical constituents should be studied deeply， and if necessary， the bioassay research should be carried out according to the main efficacy or indication of famous classical formulas. Finally， for the special processing of some herbal medicines， it is difficult to formulate the processing method， technology and standard of prepared slices. It is suggested that the scientific connotation and historical evolution of the special processing method should be thoroughly sorted out， and its technological characteristics are summarized， the modern processing technology and production processes are simulated， and then the corresponding processing methods and quality standards are formulated.
Keywords：traditional Chinese medicine;famous classical formulas;reference sample;ancient and modern dose conversion;quality consistency evaluation;processing methods
Abstract："Four in one" is a research idea of identifying the classic prescriptions from the following four dimensions： "nature， location， tendency， and syndrome". The multi-dimensional analysis of the mechanism of classic prescription Zhigancao Tang in treating coronary heart disease helps to understand the syndrome differentiation and treatment thoughts of ZHANG Zhong-jing. The coronary heart disease results from deficiency. The efficacy of Zhigancao Tang in treating coronary heart disease can be elucidated from the "nature，location，tendency， and syndrome". In terms of nature， Zhigancao Tang is pungent and sweet in flavor and warm in property， with the sweet responsible for tonifying deficiency， the pungent for dispersing Yang， resolving Yin， and eliminating surplus pathogen， and the warm for moving Yangqi， nourishing blood， and promoting blood circulation. In terms of location， Zhigancao Tang mainly acts on vessels for restoring the normal circulation of blood in the vessels and improving coronary artery stenosis and the resulting ischemia and anoxia. In terms of tendency， Zhigancao Tang tends to affect the upper and inner parts of the body to tonify deficiency in Zangfu organs， promote fluid production， nourish nutrient blood， and dissipate cold simultaneously， thus alleviating chest impediment. In terms of syndrome， Zhigancao Tang is applicable to fluid exhaustion with blood dryness and Yin-yang-qi-blood deficiency syndrome， manifested as regularly or irregularly intermittent pulse and severe palpitation. Zhigancao Tang has been widely used for the treatment of over 70 diseases classified into 10 systems， especially the cardiovascular diseases， in traditional Chinese medicine （TCM） and western medicine. As the “one” of “four in one”， Zhigancao Tang is composed of multiple Chinese herbs and its therapeutic effect is superior to the sum of its parts. It ameliorates the coronary heart disease by resisting inflammation， protecting against ischemia-reperfusion injury， adjusting the ion channels of myocardial cells， and participating in atrial remodeling and hematopoiesis. Its mechanism and clinical efficacy in the treatment of coronary heart disease have been verified by clinical and experimental studies. The utilization of the thought of "four in one" to analyze classical prescriptions enables the combination of prescriptions with syndromes， which is of great significance to the clinical application and modern development of classical prescriptions.
Keywords：four in one;Zhigancao Tang;coronary heart disease;famous classical formulas
Abstract：Metastasis is the main cause of poor prognosis of malignant tumors， and intervention with metastasis is the key measure in the treatment of malignant tumors. Hematogenous metastasis， the most common tumor metastasis， falls into the category of "Chuanshe" in traditional Chinese medicine （TCM）， with Qi deficiency and blood stasis as the critical pathogenesis. In the fight against malignant tumors， TCM emphasizes the reinforcement of healthy Qi and the elimination of pathogenic factors， exhibiting its action advantages of multiple targets， multiple mechanisms， and multiple levels. Extensive clinical evidence has shown the exact efficacy of Chinese herbal compounds designed for invigorating Qi and activating blood in delaying the progression of tumor disease and prolonging the survival period of patients. In view of the important role of hematogenous metastasis in the prognosis of tumors， more and more studies have explored the effects and mechanisms of Chinese herbal compounds capable of invigorating Qi and activating blood in intervening in hematogenous metastasis. This paper summarized the relevant literature reports in China and abroad on the intervention of Chinese herbal compounds capable of invigorating Qi and activating blood in the hematogenous metastasis of malignant tumors， in order to provide a theoretical basis for the clinical application of Qi-invigorating and blood-activating therapy in the treatment of malignant tumors. It has been found that Chinese herbal compounds formulated for invigorating qi and activating blood are effective in hindering several key steps in hematogenous metastasis through various mechanisms， including regulating the expression of cell adhesion molecules， inhibiting extracellular matrix degradation and angiogenesis， enhancing the killing effect of immunity， and improving blood hypercoagulability and hyperviscosity. Furthermore， the combination of invigorating Qi and activating blood targets the pathogenesis essence （Qi deficiency and blood stasis， characterized by sthenia in origin and asthenia in superficiality） of malignant tumor much better. Some comparative studies have demonstrated that the anti-metastasis effect of Qi-invigorating and blood-activating therapy is significantly stronger than that of the Qi-invigorating or blood-activating therapy alone， and such combination avoids the possible risk of the metastasis of malignant tumors triggered by the use of either of them. This study has provided some reference for the current clinical application of TCM for improving the prognosis of malignant tumors.
Keywords：malignant tumors;hematogenous metastasis;Qi deficiency and blood stasis;invigorating Qi and activating blood;Chinese herbal compounds
Abstract：In recent years， with the changes of people's life rhythm and living environment， the incidence of gastric ulcer has shown an increasing trend year by year， and the affected population has become younger and younger. In order to further explore the pathogenesis of gastric ulcer and its diagnosis and treatment methods， a variety of animal models of gastric ulcer have been established clinically， such as stress type， chemical factor type， pyloric ligation type， helicobacter infection type and disease-syndrome combination type. The authors intend to summarize the modeling methods and advantages and disadvantages of existing models on the basis of reviewing the etiology， pathogenesis and diagnostic criteria of gastric ulcers. It was found that the non-injurious stress method （restraint stress， restraint immersion stress and restraint freezing stress， etc.）+traditional Chinese medicine （TCM） syndrome modeling， acetic acid gavage method+TCM syndrome modeling were ideal choices for replicating animal models of acute and chronic gastric ulcer. At the same time， the analysis of the coincidence degree between each gastric ulcer model and the clinical disease characteristics of Chinese and western medicine showed that the coincidence degree of western medicine diagnostic criteria was higher than that of TCM diagnostic criteria. The successful judgment of the model was also based on western medicine diagnosis. In short， the model is insufficient in depth and breadth. It only detects a few core indicators and main indicators， ignoring the impact of secondary indicators on the diagnosis of the disease. There is also a big gap between the disease-syndrome combination model and the TCM clinical syndromes of this disease. Therefore， the depth and width of the model evaluation criteria should be strengthened， and the evaluation system of the disease-symptom combination model should be improved， in order to provide a more accurate reference for the replication of gastric ulcer models， and to replicate animal models of gastric ulcer with high coincidence degree of Chinese and western medicine for research purposes.
Keywords：gastric ulcer;animal model;integrated traditional Chinese and western medicine;clinical symptoms;stress ulcer;combination of disease and syndrome;non-injurious stress method
Abstract：Parkinson's disease （PD） is a neurodegenerative disease that seriously endangers the health of the middle-aged and elderly people. The main clinical manifestations include motor symptoms such as bradykinesia， static tremor， and myotonia and non-motor symptoms like constipation， mental disorders， sleep disorders， and autonomic nervous dysfunction. Its etiology and pathogenesis have not been fully understood， and the clinical efficacy is not satisfactory. By searching the relevant literature in China and abroad in recent years， this paper summarized the etiology， pathogenesis， and treatment of PD in both traditional Chinese medicine （TCM） and western medicine as well as the integrated TCM and Western medicine treatment. In general， liver and kidney deficiency is recognized by domestic experts in related fields as the main pathogenesis of PD. The abnormal aggregation of α-synuclein， oxidative stress， mitochondrial dysfunction， ubiquitin-proteasome system dysfunction， neuroinflammation， autophagy， microbiota-gut-brain axis regulation， and excitatory neurotoxicity are closely related to the pathogenesis of this disease. At present， treatment based on syndrome differentiation， empirical formulae from famous doctors， single Chinese herbs， and acupuncture and moxibustion are mainly adopted for the tackling of PD in TCM. Western medicine is still dominated by drug replacement therapy， supplemented by such surgical treatments as traditional immunotherapy， neurotrophic factors， and deep brain stimulation （DBS）， rehabilitation and exercise therapy， and scientific nursing. Gene therapy has become a new technical means for the treatment of this disease in recent years. In addition， the combined therapy of TCM and Western medicine has received increasing importance. This paper reviewed the pathogenesis and treatment of PD in TCM and Western medicine， so as to provide reference for its clinical diagnosis and treatment.
Keywords：Parkinson's disease;traditional Chinese and Western medicine;progress;review