Abstract：ObjectiveA comprehensive and in-depth analysis method for identification of chemical constituents in Suanzaoren Tang granules was established.MethodUltra-performance liquid chromatography-quadrupole-time of flight mass spectrometry （UPLC-Q-TOF-MS/MS） was employed with the mobile phase of 0.1% formic acid aqueous solution （A）-acetonitrile （B） for gradient elution （0-8 min， 5%-17%B； 8-10 min， 17%B； 10-11 min， 17%-18%B； 11-12 min， 18%-20%B； 12-17 min， 20%-23%B； 17-22 min， 23%-33%B； 22-30 min， 33%-60%B； 30-32 min， 60%-100%B； 32-36 min， 100%B）， the flow rate of 0.3 mL·min-1 and electrospray ionization （ESI）. High quality MS/MS data were scanned in positive and negative ion modes with scanning range of m/z 50-1 500. The local database of the chemical components from different Chinese medicines in Suanzaoren Tang granules was established by SCIEX OS software. Then the chemical components in Suanzaoren Tang granules were characterized by matching with the local database and comparing with the reference substance and literature information.ResultA total of 134 compounds were characterized and identified under positive and negative ion modes， mainly including flavonoids， triterpenoids， phthalides， steroidal saponins， alkaloids and organic phenolic acids. In addition， the sources of Chinese medicines for all compounds identified in Suanzaoren Tang granules were assigned. Among them， 41 were from Ziziphi Spinosae Semen， 11 were from Poria， 22 were from Anemarrhenae Rhizoma， 28 were from Chuanxiong Rhizoma and 35 were from Glycyrrhizae Radix et Rhizoma.ConclusionThe method can be used to identify the chemical constituents in Suanzaoren Tang granules systematically， quickly and accurately， which can provide a new strategy for the rapid and accurate identification of other Chinese patent medicines.
Keywords：Suanzaoren tang;granules;ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry （UPLC-Q-TOF-MS/MS）;qualitative analysis;chemical constituents;quality evaluation;famous classical formulas
Abstract：Huaihuasan， first recorded in Puji Benshifang by XU Shu-wei from the Southern Song dynasty， consists of four herbs， namely Sophorae Flos （fried）， Platycladi Cacumen （smashed with pestle and baked）， Schizonepetae Spica， and Aurantii Fructus （cut into pieces after the removal of pulp and then fried yellow with wheat bran）. At present， Huaihuasan and its modified formulas vary in clinical usages and dosages， and the resulting outcomes have been investigated. However， there are few reviews uncovering its historical evolution. On this basis， this review systematically combed and verified the historical evolution， dose conversion between ancient and modern times， efficacy， and indications of Huaihuasan， as well as the origin and processing of the contained herbs. The findings have demonstrated that Huaihuasan is composed of four herbs， with the original plants and medicinal parts basically the same as those recorded in the 2020 edition of Chinese Pharmacopoeia. Sophorae Flos refers to the dried flower and bud of Sophora japonica in family Leguminosae， Platycladi Cacumen the dried branch and leaf of Platycladus orientalis in family Cupressaceae， Schizonepetae Spica the dried flower spike of Schizonepeta tenuifolia in family Labiatae， and Aurantii Fructus the dried immature fruit of Citrus aurantium in family Rutaceae or its cultivated varieties. On account of the efficacy in clearing intestine， stopping bleeding， dispersing wind， and moving Qi， Huaihuasan has been mainly used to treat intestinal diseases such as bloody defecation and perianal abscess. In modern clinical practice， it is mainly applicable to patients with hematochezia and bleeding due to internal hemorrhoid， ulcerative colitis， or anal fissure. It was suggested that the raw medicinal materials should be decocted and processed according to the methods described in the 2020 edition of Chinese Pharmacopoeia. Following the conversion formula of one Qian=4 g， this paper determined the dosages of Sophorae Flos （fried）， Platycladi Cacumen （smashed with pestle and baked）， Schizonepetae Spica， and Aurantii Fructus （cut into pieces after the removal of pulp and then fried yellow with wheat bran） all to be 2 g， with the total dosage being 8 g. Such comprehensive analysis based on ancient books and modern literature has provided a more scientific reference for the clinical application， research， and development of this classical formula.
Keywords：famous classical formulas;Huaihuasan;historical evolution;textual research;efficacy and indications
Abstract：ObjectiveTo explore the inhibitory effect of dihydroartemisinin （DHA） on the proliferation of HepG2 cells， elucidate the mechanism from the perspectives of oxidative damage and energy metabolism， and discuss the possibility of combined use of DHA with sorafenib （Sora）.MethodCell counting kit-8 （CCK-8） assay was used to obtain the 50% inhibitory concentration （IC50） of DHA and Sora on HepG2 and SW480 cells and Chou-Talalay method was used to obtain the combination index （CI） of DHA and Sora. HepG2 cells were classified into the control group， DHA group （10 µmol·L-1）， Sora group （5 µmol·L-1）， and DHA + Sora group （DHA 10 µmol·L-1， Sora 5 µmol·L-1） and then incubated with corresponding drugs for 8-12 h. Seahorse XF glycolytic rate assay kit and cell mito stress test kit were employed to respectively detect the glycolysis function of cells and oxidative phosphorylation function of mitochondria. DCFH-DA and lipid peroxidation MDA assay kit were separately used to analyze the intracellular levels of reactive oxygen species （ROS） and malondialdehyde （MDA）. Western blot was applied to determine the intracellular levels of heme oxygenase-1 （HO-1） and glutamate-cysteine ligase catalytic subunit （GCLC）.ResultCompared with the control group， DHA alone inhibited the ATP synthesis in mitochondrial oxidative phosphorylation and glycolysis （P<0.01）， increased the levels of intracellular ROS and MDA （P<0.05）， and decreased the levels of HO-1 and GCLC （P<0.05） in HepG2 cells. DHA and Sora had synergistic inhibitory effect on proliferation of HepG2 and SW480 cells， with CI < 0.90. The DHA + Sora group showed stronger suppression of ATP synthesis in mitochondrial oxidative phosphorylation and glycolysis （P<0.01）， higher levels of intracellular ROS and MDA （P<0.01）， and lower levels of intracellular antioxidation-related proteins HO-1 and GCLC in HepG2 cells （P<0.01） than the DHA group.ConclusionDHA may increase the level of MDA by reducing HO-1 and GCLC and increasing ROS in HepG2 cells， which results in mitochondria oxidative damage， restricts cell glycolysis and mitochondrial oxidative phosphorylation， and thus finally inhibits the proliferation of HepG2 cells. DHA and Sora have synergistic inhibitory effect on the proliferation of HepG2 and SW480 cells， and the mechanism may be related to the synergistic oxidative damage that affects the mitochondrial electron transport chain and suppresses cell energy metabolism.
Abstract：ObjectiveTo explore the antidepressant mechanism of Yinxing Mihuan oral solution （YMO） by investigating its effect on depression model rats.MethodThe depression rats were induced by isolation combined with chronic unpredictable mild stress （CUMS） and then randomly divided into model group， fluoxetine group （10 mg·kg-1） and high-dose （618 mg·kg-1） and low-dose （309 mg·kg-1） YMO groups. A blank control group was also set up and ten rats were included in each group. Modeling lasted for 21 consecutive days， and rats were administered the 8th day after stimulation at a dose of 10 mL·kg-1 for 14 days， except those in the blank control and model groups which were given distilled water. Afterward， the sucrose preference test， open field test， tail suspension test were carried out. The pathological changes of hippocampus in depression rats were observed after hematoxylin-eosin （HE） staining. The content of interleukin-1β （IL-1β）， interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α） in the hippocampus of rats in each group and the expression of NOD-like receptor 3 （NLRP3） and other proteins in its related activation signaling pathways were detected with multi-factor detection （Luminex） and Western blot.ResultAfter 14 days of continuous administration， compared with the blank control group， the model group witnessed significantly reduced sugar water consumption rate and the times of rearing and significantly prolonged cumulative time of immobility during tail suspension （P<0.05， P<0.01）. Compared with the model group， the fluoxetine group and the high-dose YMO group saw increases in the times of rearing， times of crossing and sugar water consumption rate and a significant decrease in the cumulative time of immobility during tail suspension （P<0.05， P<0.01）. The results of HE staining showed that the neurons in the hippocampus of rats in the high-dose YMO group were arranged in order and slightly loosened， without obvious microglia infiltration observed. The levels of IL-1β， IL-6 and TNF-α in the hippocampus of the model group increased significantly as compared with the blank control group （P<0.05， P<0.01）， and their content in the high-dose YMO group was significantly lowered in the comparison with the model group （P<0.05， P<0.01）. Molecular biology experiments demonstrated that compared with the results of blank group， the expression of purinergic receptor P2X7 （P2RX7）， NLRP3， apoptosis-associated speck-like protein （ASC）， Caspase-1 and IL-1β remarkably increased in the model group （P<0.05， P<0.01）. Additionally， the expression of P2RX7， NLRP3， ASC， Caspase-1 and IL-1β was significantly inhibited in the fluoxetine group and the high-dose YMO group compared with the model group （P<0.05， P<0.01）.ConclusionYMO can improve the depression-like behaviors of rats induced by isolation combined with CUMS， and its mechanism of action is related to the regulation of the P2RX7/NLRP3 signaling pathway.
Abstract：ObjectiveTo observe the effect of total flavonoids from Epimedii Folium （TEF） on the angiogenesis of ischemic myocardium in rats after acute myocardial infarction （AMI） and discuss its molecular biological mechanism of attenuating myocardial ischemia and improving cardiac function.MethodAMI in rats was induced through the ligation of left anterior descending coronary artery. All male SD rats were randomized into sham-operated group， model group， diltiazem group （10 mg·kg-1·d-1）， and TEF low-dose and high-dose groups （100 and 200 mg·kg-1·d-1）， with 8 rats in each group. After modeling， rats in the diltiazem group and TEF groups were given corresponding doses of diltiazem and TEF， respectively， and those in the model group and sham-operated group received normal saline of equivalent volume， once a day for 7 days. After the administration， VisualSonics Vevo2100 imaging system was used to detect the cardiac structure and function and hematoxylin-eosin （HE） staining to observe the histomorphological changes in myocardial ischemic area. Immunohistochemistry was employed to analyze the expression of CD31 and α-smooth muscle actin （α-SMA） in ischemic myocardium and Western blot to detect the expression of vascular endothelial growth factor-receptor 2 （VEGF-R2） and phosphorylation of protein kinase B （Akt） in ischemic myocardium. Real-time PCR was applied to quantify the mRNA levels of vascular endothelial growth factor （VEGF） and basic fibroblast growth factor （bFGF）.ResultCompared with the sham-operated group， the model group demonstrated significant increase in left ventricular systolic diameter （LVIDs）， left ventricular internal diameter at end-diastole （LVIDd）， left ventricular end-systolic volume （LVEVs）， and left ventricular end-diastolic volume （LVEVd）， significant decrease in End-systolic thickness of left ventricular anterior wall （LVAWs）， end-diastolic thickness of left ventricle anterior wall （LVAWd）， end systolic thickness of left ventricular posterior wall （LVPWs）， stroke volume （SV）， ejection fraction （EF）， fractional shortening （FS）， and cardiac output （CO）， obvious pathological changes in the ischemic myocardium， and plummet of the expression of CD31 and α-SMA （P<0.01）， Akt phosphorylation level， protein level of VEGF-R2， and mRNA levels of VEGF and bFGF （P<0.05， P<0.01）. High-dose TEF significantly alleviated the pathological changes of ischemic myocardium as compared with the model group. Moreover， TEF high-dose group showed significantly lower levels of LVIDs， LVIDd， LVEVs， and LVEVd， significantly higher levels of LVAWs， LVAWd， LVPWs， SV， EF， FS， and CO， higher expression of CD31 and α-SMA （P<0.05， P<0.01）， and higher levels of VEGF-R2 protein， phosphorylated Akt， and VEGF and bFGF mRNA than the model group （P<0.05， P<0.01）.ConclusionTEF can effectively improve myocardial perfusion in peri-myocardial infarction area and attenuate ventricular remodeling and heart failure after AMI by up-regulating the expression of bFGF， VEGF， and VEGF-R2 in ischemic myocardium following AMI and activating phosphatidylinositol 3-kinases （PI3K）/Akt/VEGF signaling transduction pathway which can promote angiogenesis in ischemic myocardium.
Keywords：total flavonoids from Epimedii Folium;acute myocardial infarction;angiogenesis;rat
Abstract：ObjectiveTo explore the effect of Gandou Fumu decoction （GDFMD） on the oxidative damage of HepG2 cells induced by CuCl2 based on the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin （PI3K/Akt/mTOR） signaling pathway.MethodCuCl2 （200 μmol·L-1） was used to induce a copper-loaded HepG2 cell model. HepG2 cells were divided into a blank group （HepG2 cells + blank rat serum）， a model group （HepG2 cells + CuCl2 + normal rat serum）， a GDFMD group （HepG2 cells + CuCl2 + GDFMD-medicated rat serum）， an inhibitor group （HepG2 cells + NVP-BEZ235 + normal rat serum）， and a GDFMD + NVP-BEZ235 group （HepG2 cells + NVP-BEZ235 + GDFMD-medicated rat serum）. ELISA method was used to determine superoxide dismutase （SOD）， glutathione peroxidase （GSH-Px） activity， and malondialdehyde （MDA） content. The expression of microtubule-associated protein 1 light chain 3 （LC3） was detected by immunofluorescence. Phospho-PI3K/PI3K （p-PI3K/PI3K）， p-Akt/Akt， p-mTOR/mTOR， Beclin-1， LC3Ⅱ/LC3Ⅰ， and p62/Actin were determined by Western blot. PI3K， Akt， mTOR， Beclin-1， LC3Ⅰ， LC3Ⅱ， p62 mRNA expression was measured by real-time polymerase chain reaction （PCR）.ResultCompared with the blank group， the model group displayed decreased activities of SOD and GSH-Px and increased content of MDA （P<0.01）. Compared with the model group， the GDFMD group showed elevated activities of SOD and GSH-Px and reduced content of MDA （P<0.05， P<0.01）， while the inhibitor group exhibited weakened GSH-Px activity and up-regulated content of MDA （P<0.05）. Compared with the blank group， the model group showed diminished expression of p-PI3K/PI3K， p-Akt/Akt， p-mTOR/mTOR， and p62， and increased expression of Beclin-1 and LC3Ⅱ/LC3Ⅰ （P<0.01）. The expression of p-PI3K/PI3K， p-Akt/Akt， p-mTOR/mTOR， and p62 was elevated， and the expression of Beclin-1 and LC3Ⅱ/LC3Ⅰ declined in the GDFMD group （P<0.05， P<0.01）， while the p-PI3K/PI3K and p-mTOR/mTOR expression was down-regulated and the Beclin-1 and LC3Ⅱ/LC3 I expression was increased in the inhibitor group （P<0.05， P<0.01） as compared with those in the model group. Compared with the GDFMD group， the GDFMD + NVP-BEZ235 group showed down-regulated expression of p-Akt/Akt and p-mTOR/mTOR and up-regulated expression of Beclin-1 and LC3Ⅱ/LC3Ⅰ（P<0.05， P<0.01）. The expression of LC3Ⅱ protein in the model group was increased （P<0.01） as compared with that in the blank group. The expression of LC3Ⅱ protein was lower in the GDFMD group than in the model group， and higher in the GDFMD + NVP-BEZ235 group than in the GDFMD group. No significant difference in the expression of PI3K， Akt， and mTOR mRNA was observed among the groups. Compared with the blank group， the model group displayed lowered expression of p62 mRNA， and elevated expression of Beclin-1， LC3Ⅰ， and LC3Ⅱ mRNA （P<0.01）. Compared with the model group， the GDFMD group exhibited increased expression of p62 mRNA， and declining expression of Beclin-1， LC3Ⅰ， and LC3Ⅱ mRNA （P<0.01）， while the inhibitor group showed increased expression of Beclin-1 mRNA （P<0.05）. The expression of Beclin-1 and LC3Ⅱ mRNA in the GDFMD + NVP-BEZ235 group was elevated （P<0.01） as compared with that in the GDFMD group.ConclusionGDFMD may inhibit the excessive autophagy and alleviate the oxidative damage of HepG2 cells induced by CuCl2， with the underlying mechanism related to the activation of PI3K/Akt/mTOR signalling pathway.
Keywords：Gandou Fumu decoction;Wilson's disease;HepG2 cells;autophagy;phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin signal pathway
Abstract：ObjectiveTo observe the effects of Yishen Tongluo prescription （YTP） on autophagy-related proteins in rats with membranous nephropathy （MN） and explore its possible molecular mechanism in protecting the kidney.MethodTwenty of 80 Sprague-Dawley （SD） rats were randomly selected as the normal control， and the rest rats were pre-immunized and injected with cationized bovine serum albumin （C-BSA） through the tail vein to induce MN. The SD rats that were successfully modeled were randomized into the model group， benazepril hydrochloride group （10 mg·kg-1）， and low- （6.61g·kg-1）， medium- （13.22 g·kg-1）， and high-dose （26.44 g·kg-1） YTP groups， and administered with the corresponding drugs by gavage， once a day， for four consecutive weeks. Then the changes in such quantitative indicators as plasma albumin （ALB）， triglyceride （TG）， total cholesterol （TC）， serum creatinine （SCr）， blood urea nitrogen （BUN）， and 24-hour urinary total protein （UTP） were detected， followed by hematoxylin and eosin （HE） staining， Masson's trichrome staining， and periodic Schiff-methenamine （PASM） staining for observing the pathological changes in kidney under the transmission electron microscope （TEM）. The deposition of immunoglobulin G （IgG） and complement 3 （C3） in the glomerulus was detected by fluorescence immunoassay. The expression levels of autophagy marker proteins Beclin-1， microtubule-associated protein light chain 3Ⅱ （LC3Ⅱ）， and p62 were measured by immunohistochemistry （IHC）， and those of related proteins in the adenosine monophosphate-activated protein kinase / mechanisic target of rapamycin/Unc-51-like kinase 1 （AMPK/mTOR/ULK1） signaling pathway were determined by Western blot assy.ResultCompared with the normal group， the model group exhibited significantly increased UTP （P<0.01） and serum TG and TC （P<0.01）， decreased ALB （P<0.01）， disordered glomerular structure， enlarged volume， thickened basement membrane， vacuolated renal tubules， excessively deposited collagen fibers and fuchsinophilic proteins， extensively fused podocyte foot processes， and diffusely deposited IgG and C3 in glomerular capillary loops. Besides， the expression levels of Beclin-1， LC3II， and phosphorylated AMPK （p-AMPK） decreased （P<0.01）， while those of p62， phosphorylated mTOR （p-mTOR）， and phosphorylated ULK1 （p-ULK1） increased （P<0.01）. The comparison with the model group revealed that the TG， TC， and UTP levels in the low-， medium-， and high-dose YTP groups and the benazepril hydrochloride group were reduced to varying degrees （P<0.05， P<0.01）， whereas the ALB level was increased （P<0.01）. There was no statistically significant difference in SCr or BUN level. The pathological damages were alleviated. The expression levels of Beclin-1， LC3Ⅱ， and p-AMPK were up-regulated （P<0.05， P<0.01）， while those of p62， p-mTOR， and p-ULK1 were down-regulated （P<0.05， P<0.01）.ConclusionYTP protects the kidney of rats with MN possibly by regulating related proteins in the AMPK/mTOR/ULK1 signaling pathway and activating the autophagy.
Keywords：Yishen Tongluo prescription;membranous nephropathy （MN）;autophagy;adenosine monophosphate-activated protein kinase /target of rapamycin/Unc-51-like kinase 1 （AMPK/mTOR/ULK1） signaling pathway
Abstract：ObjectiveTo investigate the effects of Guiqi Dingnian prescription （GDP） on the expression of related molecules in Janus tyrosine kinase 2 （JAK2）/signal transducer and activator of transcription （JAK2/STAT） signaling pathway of D-galactose （D-gal）-induced senescent mesangial cells.MethodThe senescent mouse mesangial cells induced by 10 g·L-1 D-gal were continuously treated with 40 mg·L-1 GDP for three days. The senescence of the treated cells was determined by senescence-associated （SA）-β-gal staining. The cell cycle was detected by flow cytometry. The cell viability was analyzed using the cell counting kit-8 （CCK-8）. The mRNA expression levels of tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， nuclear transcription factor-κB （NF-κB）， and IL-1α were detected by real-time polymerase chain reaction （Real-time PCR）. The protein expression levels of STAT1， phosphorylated STAT1 （p-STAT1）， STAT3， and p-STAT3 in the JAK2/STAT signaling pathway were determined by Western blot.ResultCCK-8 results showed that the optimal concentration of GDP was 40 mg·L-1. Compared with the blank group， the positive rate of SA-β-gal in the model group was significantly higher（P<0.01）， the percentage of cells in G0/G1 phase was significantly increased（P<0.05）， the percentage of cells in G2/M and S phase was significantly decreased（P<0.01）. The mRNA expression levels of TNF-α，IL-6，NF-κB and IL-1α were significantly increased（P<0.01）. Compared with the model group， the model + GDP group exhibited significantly decreased SA-β-gal-positive cells （P<0.05）， reduced cells in the G0/G1 phase （P<0.05）， increased cells in the G2/M and S phases （P<0.01）， and down-regulated TNF-α， IL-6， NF-κB， and IL-1α mRNA expression （P<0.05） and STAT1， p-STAT1， STAT3， and p-STAT3 protein expression （P<0.05）.ConclusionGDP delays the senescence of mouse mesangial cells possibly by down-regulating the expression of related molecules in the JAK2/STAT pathway.
Keywords：Guiqi Dingnian prescription （GDP）;Janus tyrosine kinase 2 （JAK2）/signal transducer and activator of transcription （JAK2/STAT） signaling pathway;mouse mesangial cells;D-galactose （D-gal）;senescence
Abstract：ObjectiveTo investigate the effects of curdione on the proliferation， apoptosis and cell cycle of triple negative breast cancer cell line MDA-MB-231.MethodMDA-MB-231 cells were cultured in vitro with capecitabine （positive control） and curdione at different concentrations （125， 250， 500， 1 000， and 2 000 μmol·L-1）， respectively， for detecting their viability using the cell counting kit-8 （CCK-8） at 24 and 48 h. Three effective inhibitory concentrations （250， 500， and 1 000 μmol·L-1） against cell proliferation were selected for subsequent experiments. The effect of curdione on cell cycle was determined by flow cytometry combined with propidium iodide （PI） staining. After the set-up of high-concentration （2 000 μmol·L-1） group， the effect of curdione on cell mitochondrial membrane potential was measured by JC-1（5，5，6，6-tetrachloro-1，1，3，3-tetraethylbenzimidazolylcarbocyanine iodide） staining， followed by the detection of cell apoptosis by flow cytometry combined with Annexin V-FITC/PI double staining. The changes in cell cycle status and apoptosis-related protein expression following curdione intervention were assayed by Western blot.ResultCompared with the blank control， curdione at 250， 500， 1 000， and 2 000 μmol·L-1 significantly inhibited the proliferation of MDA-MB-231 cells （P<0.01）， exhibiting a concentration- and time-response relationship. The half maximal inhibitory concentration （IC50） values at 24 and 48 h were 1 607 and 1 401 μmol·L-1， respectively. Curdione at 250， 500， and 1 000 μmol·L-1 arrested cells in G1 phase. Curdione at 250 μmol·L-1 had no effect on cell mitochondrial membrane potential， which， however， declined significantly in the 500， 1 000， and 2 000 μmol·L-1 groups （P<0.05， P<0.01）. Curdione at 250， 500， and 1 000 μmol·L-1 obviously increased the proportion of apoptotic cells （P<0.05， P<0.01）. Curdione at each concentration elevated the Bcl-2-associated X protein （Bax）/B-cell lymphoma 2 （Bcl-2） ratio （P<0.05， P<0.01）， but did not change the cysteinyl aspartate-specific protease-3 （Caspase-3） expression. The protein expression levels of Caspase-9， cleaved Caspase-9， cleaved Caspase-3， p53， and p21 were up-regulated （P<0.05）.ConclusionA certain concentration of curdione inhibits the proliferation of MDA-MB-231 cells， which may be related to its efficacy in arresting cell cycle and inducing apoptosis.
Keywords：curdione;triple negative breast cancer;MDA-MB-231 cells;cell cycle;apoptosis
Abstract：ObjectiveTo investigate the effect of allicin （ALL） on learning and memory ability of rats with vascular dementia （VD） and the possible mechanism.MethodThe VD rats induced by modified bilateral common carotid artery occlusion （BCCAO） were randomly divided into the VD group， low- and high-dose ALL （ALL-L and ALL-H） groups， and the sham operation （S） group， with 15 rats in each group. In the ALL-L and ALL-H groups， ALL was injected into the femoral vein at 5 mg·kg-1 and 20 mg·kg-1， respectively， while the same volume of normal saline was injected in the S and VD groups， once a day， for two successive weeks. Morris water maze （MWM） was used to test the learning and memory ability of rats. Hematoxylin and eosin （HE） staining was conducted to observe the pathological changes in hippocampal tissue， followed by the detection of inflammatory factors tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and IL-1β as well as oxidative stress indexes malondialdehyde （MDA）， superoxide dismutase （SOD）， and glutathione peroxidase （GSH-Px） in rat hippocampus. The apoptosis of hippocampal cells was detected by TdT-mediated dUTP Nick end Labeling（TUNEL） assay. The expression levels of apoptosis and autophagy-related proteins cysteinyl aspartate-specific protease-3 （Caspase-3）， B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated X protein （Bax）， microtubule-associated protein light chain 3Ⅱ （LC3Ⅱ）， LC3Ⅰ， and the mammalian homolog of yeast ATG6 （Beclin 1） in hippocampus were determined by Western blot.ResultThe comparison with the VD group revealed that the learning and memory abilities of rats in the ALL-H and ALL-L groups were significantly improved （P<0.05）. The TNF-α， IL-6， IL-1β， and MDA levels in hippocampus were lowered （P<0.05）， whereas the SOD and GSH-Px activities were enhanced （P<0.05）. The apoptosis rates were declined （P<0.05）， with an even lower rate noticed in the ALL-H group （P<0.05）. The expression levels of Caspase-3， Bax， LC3Ⅱ/LC3Ⅰ ratio， and Beclin-1 in the ALL-H and ALL-L groups were significantly down-regulated in contrast to those in the VD group （P<0.05）， while that of Bcl-2 was up-regulated （P<0.05）. The ALL-H group exhibited better performances than the ALL-L group （P<0.05）.ConclusionALL could improve the learning and memory ability of VD rats to some extent， which may be attributed to its inhibition against inflammatory reaction， oxidative stress， and neuronal apoptosis and autophagy.
Abstract：ObjectiveTo explore the distribution characteristics of traditional Chinese medicine （TCM） syndromes of abnormal uterine bleeding associated with ovulatory dysfunction （AUB-O）.MethodA clinical and epidemiological investigation was conducted in 6 688 patients with AUB-O from the provincial， municipal， and county/district hospitals in 29 provinces， autonomous regions， and municipalities across China to identify the distribution characteristics of their TCM syndromes.ResultThe AUB-O patients nationwide were mainly differentiated into the kidney Qi deficiency syndrome （17.34%）， the spleen Qi deficiency syndrome （13.25%）， the Qi and blood deficiency syndrome （12.62%）， the Qi deficiency and blood stasis syndrome （8.45%）， and the kidney Yin deficiency syndrome （6.88%）. AUB-O resulted from Qi deficiency， Yin deficiency， and blood deficiency， often involving the kidney， spleen and liver. The analysis of the regional distribution of TCM syndromes in AUB-O patients revealed that kidney Qi deficiency， spleen Qi deficiency， and Qi and blood deficiency were the shared syndromes. However， due to regional discrepancy， the TCM syndrome varied widely from one geographic region to another. The kidney Qi deficiency syndrome was more frequently seen in North China， Northwest China， Southwest China， and East China， but less frequently in central China， Northeast China， and South China. The spleen Qi deficiency syndrome occurred most frequently in central China， while the East China had the highest frequency of Qi and blood deficiency syndrome. The spleen and kidney deficiency syndrome was mostly present in central China， North China， and Southwest China， the Qi deficiency （kidney deficiency） and blood stasis syndrome in Northwest China， South China， and North China， the kidney Yin deficiency syndrome in East China， Northwest China， and Northeast China， the deficiency-heat syndrome in Southwest China and East China， the kidney Yang deficiency syndrome in South China， the liver depression and blood heat syndrome in Northeast China， and the liver depression and spleen deficiency syndrome in central China. The diagnostic criteria for the kidney Qi deficiency， spleen Qi deficiency， Qi and blood deficiency， Qi deficiency and blood stasis， and kidney Yin deficiency syndromes were not significantly different from the previous ones. The distinctive symptoms for the kidney Qi deficiency syndrome were irregular vaginal bleeding， heavy menstrual flow， or shortened menstrual cycle， back soreness and pain， and forgetfulness， while those for the spleen Qi deficiency syndrome mainly included the shortened menstrual cycle， mental fatigue， lack of strength， poor appetite， loose stool， and white tongue coating. The Qi and blood deficiency syndrome were mainly manifested as the shortness of breath， laziness to speak， pale complexion， dizziness， and palpitation. The Qi deficiency and blood stasis syndrome were mainly judged by the scanty menstrual flow frequently or occasionally accompanied by blood clots， mental fatigue， lack of strength， and dark purple tongue. The ovulatory bleeding， dizziness， tinnitus， vexing heat in chest， palms and soles， and night sweat were the characteristic signs for the kidney Yin deficiency syndrome.ConclusionThere exist certain rules in the geographical distribution of TCM syndromes of AUB-O patients， which has provided a reference for the clinical treatment of AUB-O in accordance with the local conditions.
Keywords：abnormal uterine bleeding associated with ovulatory dysfunction （AUB-O）;traditional Chinese medicine （TCM） syndrome;epidemiological investigation
Abstract：ObjectiveTo evaluate the efficacy of modified Xiaoji Baozhong granules combined with scraping in the treatment of abdominal obesity with gastric heat and dampness syndrome and its effect on adipocytokines.MethodOne hundred and forty-four patients were randomly divided into the control group and the observation group， 72 cases in each group. The lifestyle adjustments were conducted in both groups. The observation group was given a comprehensive traditional Chinese medicine regimen of modified Xiaoji Baozhong granules combined with scraping. The control group was given modified Xiaoji Baozhong granules combined with scraping. The course of treatment was three months. The body mass index （BMI）， body fat percentage （FP）， waist circumference （WC）， waist to hip ratio （WHR）， obesity and abdominal fat thickness of the two groups were compared before and after treatment. The levels of fasting blood glucose （FBG）， insulin resistance index （HOMA-IR）， high density lipoprotein （HDL）， low density lipoprotein （LDL）， triglyceride （TG）， total cholesterol （TC）， adiponectin， leptin， resistin， endolipid and tumor necrosis factor-α （TNF-α） were compared between the two groups before and after treatment. The clinical efficacy and safety were evaluated.ResultAfter treatment， the BMI， FP， obesity， WC and WHR of the observation group were lower than those of the control group （P<0.01）. After treatment， the abdominal wall fat thickness and intra-abdominal fat thickness of patients in the observation group were lower than those in the control group （P<0.01）. The FBG， HOMA-IR， TG， TC and LDL of the observation group were lower than those of the control group （P<0.01）， while the HDL level was higher than the control group （P<0.01）. Adiponectin in the observation group was higher than the control group after treatment （P<0.01）. The leptin， resistin， visfatin and TNF-α were lower than those in the control group （P<0.01）. The total clinical efficacy rate of patients in the observation group was 88.24% （60/68）， which was better than 73.53% （50/68） in the control group （χ2=4.755， P<0.05）.ConclusionIn addition to lifestyle adjustments， modified Xiaoji Baozhong granules combined with scraping treatment can reduce the degree of obesity and fat thickness in patients with abdominal obesity， and regulate lipid， carbohydrate metabolism and adipocytokines， with a better clinical efficacy than simple scraping therapy and the safety in practice.
Keywords：abdominal obesity;stomach heat and dampness syndrome;Xiaoji Baozhong granules;scraping;traditional Chinese medicine comprehensive therapy;adipocyte factor
Abstract：ObjectiveTo investigate the efficacy of Baxian Xiaoyaotang （BXT） in treating ankylosis of wind-cold-dampness obstruction syndrome after acute Achilles tendon rupture surgery and its effects on transforming growth factor-β1 （TGF-β1）， insulin-like growth factor-1 （IGF-1）， and epidermal growth factor （EGF）.MethodAccording to the visiting sequence， 66 patients with fresh closed Achilles tendon rupture were included and randomly divided into a treatment group （n=33） and a control group （n=33）. Patients in both groups underwent surgical repair， followed by immobilization in long-leg brace， which was then replaced by the boot brace in the fourth week， with the plantar-flexion angle adjusted correspondingly. Six weeks later， the brace was removed for accelerated functional rehabilitation training. On this basis， patients in the treatment group were further instructed to fumigate and wash the affected Achilles tendon with BXT， twice a day， for 45 d. The Leppilahti Achilles tendon performance scores and American Orthopaedic Foot and Ankle Society （AOFAS） ankle-hindfoot scores between the two groups were compared at the time of brace removal and the third， sixth， and twelfth months after surgery. The strength of triceps surae on the affected side was evaluated at the last follow-up visit. The serum TGF-β1， IGF-1， and EGF levels were detected before and after treatment. The wind-cold-dampness obstruction syndrome scores， symptom scores， the changes in foot dorsiflexion angle， and the overall clinical efficacy were compared.ResultThe changes in scores of patients receiving different treatment measures did not synchronize. After the removal of brace， the Leppilahti Achilles tendon performance score and AOFAS ankle-hindfoot score determined at three time points in the treatment group were higher than those in the control group （P<0.05）. At the last follow-up visit， the good-to-excellent rate of muscle strength in the treatment group was 93.94% （31/33）， higher than 72.73% （24/33） in the control group （χ2=0.031，P<0.05）， implying that the strength of triceps surae in the treatment group was better recovered. After treatment， the serum TGF-β1， IGF-1， and EGF levels in both groups were increased in contrast to those before treatment （P<0.05）， and these levels in the treatment group were all higher than those in the control group （P<0.05）. The foot dorsiflexion angle and the wind-cold-dampness obstruction syndrome score in the treatment group were superior to those in the control group （P<0.05）. The overall response rate of the treatment group was 90.91% （30/33）， higher than 75.76% （25/33） of the control group （χ2=6.981， P<0.05）. No adverse reactions occurred during the treatment.ConclusionThe external fumigation and washing with BXT alleviates both the clinical symptoms and traditional Chinese medicine （TCM） syndrome， improves the joint function score， triceps surae strength， and other indicators， elevates the serum TGF-β1， IGF-1， and EGF levels， and enhances the strength and toughness of Achilles tendon of patients with ankylosis due to wind-cold-dampness obstruction after the acute Achilles tendon rupture surgery. Its clinical efficacy is better than that of functional rehabilitation training.
Abstract：ObjectiveThis study aims to investigate the effect of addition and subtraction treatment to Renshen Maidongsan on metabolism of glucose， lipid and pregnancy outcome when used to gestational diabetes mellitus （GDM） with deficiency of both Qi and Yin.MethodIn this study， 108 patients with GDM were randomly divided into observation group with 54 cases and control group with 54 cases. Patients in control group took Yuquan wan orally， 6 g/time， 4 times/day ， and patients in observation group were prescribed addition and subtraction treatment to Renshen Maidongsan， 1 dose/day. The course of treatment for both groups was 4 weeks. Oral glucose tolerance test （OGTT） was performed before and after treatment. Fasting blood glucose （FBG） ， 1 h our plasma glucose （1 h PG）， 2 h our plasma glucose （2 h PG）， glycosylated hemoglobin （HbA1c） and other indicators were detected to record the pregnancy outcome. Also， the levels of total cholesterol （TC）， triglyceride （TG）， high density lipoprotein cholesterol （HDL-C）， low density lipoprotein cholesterol （LDL-C）， adiponectin， serum amyloid A （SAA）， tumor necrosis factor-α （TNF-α）， resistin， interleukin-1β （IL-1β） and interleukin-6 （IL-6） were detected before and after treatment. Then the safety evalution was carried out.ResultThe levels of insulin resistance index （HOMA-IR）， FBG， 1 h PG， 2 h PG， HbA1c， TC， LDL-C， TG， IL-6， IL-1β， TNF-α， SAA， leptin and resistin in observation group were lower than the data checked in control group （P<0.01）， and the levels of HOMA-β， insulin sensitivity index （InISI） and adiponectin were higher than the standards of from control group （P<0.01）. Then the incidence of medical cesarean section， pregnancy induced hypertension， polyhydramnios， premature rupture of membranes， premature delivery and adverse pregnancy outcomes were lower than those in control group （P<0.01）.ConclusionAddition and subtraction treatment to Renshen Maidongsan can improve insulin sensitivity and islet β-cell function， regulate inflammatory factors and adipokines， improve insulin resistance （IR）， effectively regulate glucose and lipid metabolism， improve maternal and infant adverse pregnancy outcomes when used on patients with GDM and deficiency of Qi and Yin， and it was safe to use in clinic.
Keywords：gestational diabetes mellitus;syndrome of deficiency of Qi and Yin;Renshen Maidongsan;inflammatory factors;adipokines;pregnancy outcome
Abstract：ObjectiveTo compare the contents of adenosine， gastrodin， p-hydroxybenzyl alcohol， p-hydroxybenzaldehyde， parisinin B and parisinin A in Chijian （the aerial part of Gastrodia elata） and Gastrodiae Rhizoma， and compare their effects on immune function and intestinal microflora， evaluating whether it is necessary to study and develop Chijian.MethodThe contents of these six constituents were determined by ultra performance liquid chromatography （UPLC）， the mobile phase was 0.1% formic acid aqueous solution （A）-acetonitrile （B） for gradient elution （0-4 min， 0.5%B； 4-5 min， 0.5%-2%B； 5-10 min， 2%-15%B； 10-12 min， 15%-20%B； 12-15 min， 20%-95%B； 15-17 min， 95%B； 17-17.5 min， 95%-0.5%B； 17.5-20 min， 0.5%B）， the flow rate was 0.5 mL·min-1， the detection wavelength was 270 nm. The difference of pharmacological activity of water extracts of Chijian and Gastrodiae Rhizoma was compared， the clearance index， corrected clearance index and peripheral blood were measured in mice model with low immune function induced by cyclophosphamide， B lymphocyte proliferation was determined by lymphocyte transformation test in vitro， intestinal microflora was analyzed by 16S rDNA technology and bioinformatics was conducted.ResultThe total contents of these six components in powder and ethanol extract of Chijian were higher than that of Gastrodiae Rhizoma， but the total contents of these six components in their water extract were similar， and the total contents of gastrodin and p-hydroxybenzyl alcohol met the requirements of the 2020 edition of Chinese Pharmacopoeia. Compared with the blank group， the clearance index of immunocompromised mice was significantly increased in the middle-dose （10 g·kg-1） group of Chijian water extract， middle- and low-dose （10， 5 g·kg-1） groups of Gastrodiae Rhizoma water extract （P<0.05）， the levels of erythrocyte and hematocrit in peripheral blood were significantly increased in the high-dose （20 g·kg-1） groups of water extracts of Chijian and Gastrodiae Rhizoma （P<0.05， P<0.01）， water extract of Gastrodiae Rhizoma with concentration of 400 g·L-1 and the water extract of Chijian with the concentration of 100 g·L-1 could promote the proliferation of B lymphocytes induced by lipopolysaccharide. Studies on intestinal microflora showed that compared with the blank group， at the phylum level， the water extracts of Chijian and Gastrodiae Rhizoma increased the relative abundance of Bacteroidetes and decreased the relative abundance of Firmicutes， at the genus level， they increased the relative abundance of Prevotellaceae_UCG-001 and Ruminococcaceae_UCG-005， and decreased the relative abundance of Anaerotruncus， unclassified_f_Erysipelotrichaceae and Candidatus_Stoquefichus. These intestinal bacteria were related to the immune system， cell proliferation， and metabolism regulation.ConclusionThe total contents of 6 components in the powder， the ethanol and the water extracts of Chijian are higher than or close to those of the corresponding samples of Gastrodiae Rhizoma， the pharmacological activity of Chijian water extract is similar to that of Gastrodiae Rhizoma water extract， indicating that Chijian is worthy of further research and development.
Abstract：ObjectiveTo analyze and identify the flavonoids of Citri Reticulatae Pericarpium with different aging time by an ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry （UPLC-Q-Orbitrap HRMS）.MethodCompounds were separated on Agilent Extend-C18 column （3.0 mm×100 mm， 1.8 μm）， mobile phase was 0.1% acetic acid aqueous solution （A）-0.1% acetic acid methanol solution （B） for gradient elution （0-25 min， 5%-95%B； 25-30 min， 95%B； 30-30.1 min， 95%-5%B； 30.1-35 min， 5%B）， the flow rate was 0.4 mL·min-1， and the column temperature was set at 30 ℃. High resolution mass spectrometry was performed with electrospray ionization （ESI）， and scanned in positive and negative ion modes by means of full scan/data dependent secondary scan （Full MS/dd-MS2）. The multistage ion fragment information combined with mzCloud network database， local high resolution mass spectrometry database of traditional Chinese medicine components （OTCML）， literature information and relevant reference materials were used for accurate qualitative analysis.ResultTotally 43 flavonoids in Citri Reticulatae Pericarpium were identified， including 24 flavones， 5 flavonols， 13 dihydroflavones and 1 chalcone. The flavonoids in samples with different aging time were basically consistent in material types， but the peak area was different. According to the comparison of relative content in the peak area， it was found that the relative contents of 30 flavonoids showed an overall increasing trend with the increase of aging time. Among them， the relative contents of 24 flavonoids （such as hesperidin， diosmin， 6-demethoxytangeretin， nobiletin and tangeretin） increased significantly. There was no significant change in the relative contents of the other 13 flavonoids （such as naringenin and neohesperidin）.ConclusionAn efficient method is established in this paper to identify flavonoids in Citri Reticulatae Pericarpium with different aging time and their relative content changes rapidly and accurately. The findings provide a methodological reference for the study on pharmacodynamic material base and quality control of Citri Reticulatae Pericarpium， and it provides experimental basis that drugs processed long time ago have better effect of Citri Reticulatae Pericarpium.
Keywords：Citri Reticulatae Pericarpium;ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry （UPLC-Q-Orbitrap HRMS）;aging time;flavonoids;electrospray ionization （ESI）
Abstract：ObjectiveTo clone uridine diphosphate （UDP）-glucose dehydrogenase （UGDH） gene of Glycyrrhiza uralensis and analyze its bioinformatics and expression.MethodTotal RNA was extracted from roots， stems， and leaves of 6-week-old seedlings of G. uralensis， the complementary deoxyribonucleic acid （cDNA） sequence of GuUGDH1 gene （Gu was short for G. uralensis） was cloned by reverse transcription polymerase chain reaction （RT-PCR）， then sequencing and bioinformatic analysis were performed， and the specificity of the tissue was analyzed by real-time fluorescence quantitative PCR （Real-time PCR）.ResultThe open reading frame（ORF）of GuUGDH1 gene was 1 443 bp in length and encoded 480 amino acid residues （GenBank accession number of MT968993）. Bioinformatics analysis showed that GuUGDH1 was a stable acidic hydrophilic protein with a relative molecular weight of 53.056 kDa， an isoelectric point of 5.89， no signal peptide and no transmembrane helix， and all of them were outside the membrane. There were three typical conserved domains， which belonged to the UDP-glucose/guanosine diphosphate （GDP）-mannose dehydrogenase family. Phylogenetic analysis showed that the GuUGDH1 gene was closely related to Glycine max and Spatholobus suberectus. The results of Real-time PCR showed that the expression of GuUGDH1 gene could be detected in the roots， stems， and leaves of 6-week-old seedlings of G. uralensis， and the expression level in the roots was significantly higher than that in the stems and leaves.ConclusionIn this study， the UGDH1 gene of G. uralensis was cloned and its protein sequence characteristics were systematically analyzed， which can provide theoretical basis for further research on the catalytic function of UGDH1 protein.
Abstract：ObjectiveTo screen out the suitable nonpolar molecular cosolvent and concentration with adventitious root phenotype and ginsenoside content in the controlled experiment as the evaluation indexes， so as to lay a solid foundation for exploring the causes for good shape and high quality of Panax quinquefolium.MethodAfter being treated with different concentrations of dimethyl sulfoxide （DMSO） and ethanol， the adventitious roots were scanned using a panoramic scanner， and the resulting images were used for measuring the branch number and average diameter by WinRHIZO Pro 2016， Synbiosis ProtoCol 3 colony counter， Image J， and SmartRoot. The contents of ginsenosides Rg1， Rb1， and Re were determined by ultra-performance liquid chromatography tandem mass spectrometry （UPLC-MS/MS）.ResultCompared with the blank control， the 0.1% DMSO and 75% ethanol made the adventitious root phenotype and ginsenoside contents significantly changed. Specifically， the branch number and average diameter were significantly reduced. The ginsenoside Rg1 in the adventitious roots decreased after 0.1% DMSO treatment， whereas the ginsenosides Rg1 and Re increased after 75% ethanol treatment. The adventitious root phenotype and ginsenoside contents in the 0.1% DMSO treatment group were not significantly different from those in the control group.ConclusionThe 0.01% DMSO does not affect the adventitious root growth of P. quinquefolium and is insoluble in water， enabling it to be considered as a suitable nonpolar molecular cosolvent for future research on the genetic causes for the good shape and high quality of P. quinquefolium.
Abstract：ObjectiveTo study the distribution characteristics of mineral elements in Gastrodia elata samples with different grades and specifications （variants） from diverse producing areas and their classification and identification evidences.MethodFourteen mineral elements in 31 batches of Gastrodia elata samples of different grades and specifications （variants） from diverse producing areas were determined by atomic absorption spectrophotometry， Mo-Sb colorimetry， and curcumin colorimetry， and then subjected to correlation analysis （CA）， discriminant analysis （DA）， and principal component analysis （PCA）.ResultThe content of K， N， and P in G. elata was the highest， enabling them to serve as the nutritional limiting factors affecting its growth. The G. elata samples could be identified by the variation trend of elements （K>N>P>Mg>Ca>Fe>B>Zn>Mn>Cu>Ni>Cr>Pb>Cd）. The comparison of G. elata samples from multiple producing areas showed that G. elata from Zhaotong has the highest P， Fe， and Cd content， that from Lijiang the highest K content， that form Luotian the highest Zn and Cr content， and that from Jinzhai the highest Cu and Pb content. The content of Mg， B， Pb， and Cr in G. elata f. elata was higher than that in G. elata f. glauca. It was found that the content of P， Cu， and Cd in commercially available G. elata products gradually increased with the decrease in the commercial grade， while that of Mg， Fe， B， and Ni mostly decreased. As revealed by CA， Fe was positively correlated with Mg， Cr， and B. The producing areas of G. elata samples could be effectively identified by DA with Cd， Pb， Cr， Cu， B， and Ni as the main variables， and the accuracy reached up to 85.71%. According to the PCA of mineral elements in G. elata f. glauca from Zhaotong， Yunnan Province， Fe， Cr， Mg， Cd， P， Mn， B， Pb， and Cu exerted a greater influence on G. elata.ConclusionThe determination of mineral elements in G. elata samples contributes to identifying their authenticity and origin due to the easy operation， accurate results， and good stability.
Keywords：Gastrodia elata;Gastrodia elata f. glauca;Gastrodia elata f. elata;specification;grade;mineral elements;content characteristics
Abstract：ObjectiveTo investigate the anti-inflammatory effects of water extract of the Iris halophila root on lipopolysaccharide（LPS） stimulated RAW264.7 cells and analyze its chemical constituents.MethodThe supernatant of YWG prepared by water extraction and alcohol precipitation was separated by AB-8 macroporous adsorption resin column chromatography to obtain ethanol eluates with different concentrations （YWG，YWG-0%，YWG-20%，YWG-40%，and YWG-60%）. Cell counting kit-8（CCK-8） assay was used to determine the effects of YWG-0%，YWG-20%，YWG-40%，and YWG-60% on the viability of RAW264.7 cells. Griess assay was employed to detect the nitric oxide （NO） level in LPS-stimulated RAW264.7 cells. The release of tumor necrosis factor（TNF）-α，interleukin（IL）-6，IL-10，and IL-1β was detected by enzyme-linked immunosorbent assay（ELISA）. YWG and the elution site with the most robust anti-inflammatory activity were identified and compared by ultra-high performance liquid chromatography-quadrupole-time of flight-mass spectrometry （UHPLC-Q-TOF-MS/MS）.ResultEthanol eluates with different concentrations inhibited the release of NO，TNF-α，IL-1β， and IL-6 in the supernatant of LPS induced RAW264.7 cells （P<0.05），and promoted the release of IL-10 （P<0.05）. YWG-60% displayed a highly significant effect （P<0.01）. A total of 127 constituents were detected from the comparison of YWG and YWG-60% by UHPLC-Q-TOF-MS/MS in the positive and negative ion modes，including 61 flavonoids. YWG-60% contained 25 flavonoids with elevated content as compared with YWG.ConclusionYWG-60% showed potent anti-inflammatory effect，and the effective anti-inflammatory constituents were presumedly flavonoids. The findings of this study are expected to provide a scientific theoretical basis for the basic research on the medicinal effect of the water extract of YWG.
Abstract：ObjectiveTo analyze the main factors affecting the Ziziphus jujuba distribution and expand the understanding of its distribution and the corresponding influencing factors by comparing the distribution sites of Z. jujuba predicted by models with those recorded in the literature.MethodMore than 200 distribution sites of Z. jujuba accompanied by 55 environmental factors were obtained from literature and specimen review. The environmental factors that affect the distribution of Z. jujuba were explored by maximum entropy （MaxEnt） model， and the potential distribution areas of Z. jujuba in China were analyzed by ArcGIS， followed by the verification of the main environmental factors using receiver-operating characteristic （ROC） curve and Jackknife method.ResultThe area under the curve （AUC） values for the test data and training data were both greater than 0.9， which perfectly satisfied the standard， indicating that the research results were accurate and reliable.ConclusionThe annual average temperature， the average temperature in May， the average temperature in the warmest season， vegetation type， soil type， average temperature in June， average temperature in September， and average temperature in August are proved to be the main environmental factors affecting the distribution of Z. jujuba， which can be found almost all over China， except for Heilongjiang and Tibet. Z. jujuba is most suitable to be planted in southeastern Sichuan， Chongqing， southern Gansu， Ningxia， most areas of central Shaanxi， eastern and southwestern Shanxi， Henan， eastern and northern Hubei， northern and eastern Anhui， Shandong， Hebei， Beijing， Tianjin， western Liaoning， and Zhejiang. As revealed by literature review， the most suitable growing areas of Z. jujuba are southeastern Sichuan， central Shaanxi， southwestern Shanxi， western and northern Henan， Shandong， and southwestern and eastern Hebei.
Keywords：Ziziphus jujuba;environmental factors;potential distribution areas;maximum entropy （MaxEnt） model;ArcGIS
Abstract：This review made a systematic textual research on the historical evolution and changes of the origin， producing areas and quality， harvest time and processing methods， and cultivation technique of Poria in famous classical formulas from the Catalogue of Ancient Famous Classical Formulas （the First Batch） by referring to the literature of ancient materia medica and medical books combining with the modern literature in CNKI， Wanfang Data and other databases， which could provide reference for the development and research of the famous classical formulas containing Poria. According to the research， the origin of Poria in ancient times was the dried sclerotia of Poria cocos. The producing area of Poria is widely distributed in China， mainly produced in Anhui， Hubei， Henan and Yunnan provinces. The quality of Poria from Yunnan is better， and the yield is larger in Anhui and Hubei provinces. The quality evaluation of Poria is basically the same from ancient to modern， which has the characteristics of large block， heavy weight， thin and wrinkled skin， and white and delicate cross section. The harvesting period of Poria is usually in August of the lunar calendar. The initial processing has undergone the evolution from drying in shade in the ancient time to drying in shade after sweating in the modern time. From ancient times to the present， Poria has been processed by cutting to prepare the Fuling lumps and Fuling slices. The cultivation technique has experienced the evolution process of "wild Poria-inoculation of live pine root-basswood cultured". At present， Poria is mainly cultivated by artificial basswood with mature technical methods and abundant yields， which can meet the research and development needs of the famous classical formulas.
Keywords：famous classical formulas;Poria;origin;textual research on materia medica;cultivation;processing;harvesting time
Abstract：In recent years， the incidence rate of andrological diseases has shown a significant growth trend. Considering the unavailability of a perfect theoretical system for andrology in traditional Chinese medicine （TCM） and the complex pathogenesis despite of the limited types of andrological diseases， it is necessary to improve the clinical efficacy of andrological diseases so as to satisfy the needs of patients. Therefore， the China Association of Chinese Medicine （CACM） organized the andrologists of TCM and western medicine and the outstanding young clinicians to discuss the andrological diseases responding specifically to TCM or integrated TCM and western medicine， such as chronic prostatitis， male infertility， benign prostatic hyperplasia， erectile dysfunction， and premature ejaculation， determine their diagnostic criteria in western medicine， and standardize the specifications for TCM diagnosis and treatment based on syndrome differentiation， thus formulating recognized and integrated diagnosis and treatment protocols. Apart from proposing suggestions on the treatment of such andrological diseases with TCM and western medicine， the experts have also figured out the andrological diseases responding specifically to TCM， the optimal intervention time of TCM and western medicine， and the suitable measures including surgery. The resulting consensus helps to better guide the formulation of accurate， personalized， and optimized treatment plans in clinical practice and improve the diagnosis and treatment effects of andrological diseases by giving full play to the advantages of TCM， which will in turn contribute to further innovation and development of TCM.
Keywords：andrology;diseases responding specifically to traditional Chinese medicine （TCM）;expert's perspectives
Abstract：Interventional or thrombolytic therapy for acute myocardial infarction （AMI） has the effect of opening occluded coronary arteries. However， due to no myocardial regurgitation， ischemia/reperfusion injury and microvascular structure and function destruction， leading to myocardial fibrosis following AMI， ventricular remodeling， and even cardiac failure may occur. Collaterals doctrine is used to guide the investigation into traditional Chinese medicine pathogenesis and clinical treatment of myocardial fibrosis following AMI. Consequently， it proposes the pathogenesis characteristic of "Qi deficiency and blood stasis， collaterals obstruction" and medication principle of "dispersing and dredging"， to develop the formula of Tongxinluo. Fundamental and clinical researches have demonstrated it can effectively protect the structure and function of myocardial microvascular endothelial cells， improve no reflow and reperfusion injury， and inhibit myocardial fibrosis after AMI， which further supports scientific value of the collaterals doctrine.
Keywords：collaterals doctrine;acute myocardial infarction （AMI）;myocardial fibrosis;Qi deficiency and blood stasis;collaterals obstruction;dispersing and dredging;Tongxinluo
Abstract：This study summarized and analyzed the modeling methods， modeling mechanisms， and advantages and disadvantages of existing animal models of acute pancreatitis （AP） with the purposes of replicating animal models more in line with the clinical syndrome characteristics of traditional Chinese medicine （TCM） and Western medicine， improving the basic research of AP， and promoting its in-depth research. Further， the clinical diagnostic criteria of TCM and Western medicine were drawn up， and the agreement was evaluated between animal models of AP and clinical syndrome characteristics of TCM and Western medicine. The results demonstrated that the AP model induced by choline-deficient ethionine-supplemented diet （CDE）-supplemented diet was highly consistent with the clinical syndromes of TCM and Western medicine， which was simple in operation and small in mortality. However， the modeling did not involve the pathogenic factors of TCM. Although many approaches have been proposed to establish animal models of AP so far， most of the models meet the diagnostic criteria of Western medicine but， lacking the manifestation of TCM symptoms， less agree with the clinical syndromes of TCM. At present， animal models of AP alone are mostly used to study the pharmacodynamics and mechanisms of Chinese and Western medicines， which fail to meet the requirements of syndrome differentiation and treatment in TCM theories. Only the animal models of AP which conform to the TCM syndrome characteristics and are basically consistent with clinical syndromes greatly contribute to the essence study of TCM syndromes and the development of innovative Chinese medicines. Therefore， establishing animal models that can simultaneously reflect the clinical syndrome characteristics of AP in both TCM and Western medicine and replicating more realistic， accurate and comprehensive animal models of AP are worthy of further research.
Keywords：acute pancreatitis;traditional Chinese medicine （TCM） and Western medicine;clinical syndrome characteristics;animal model
Abstract：Liver， as a critical organ of metabolism and detoxification， can be damaged by viral infection， drug abuse， and heavy drinking. Liver diseases pose a serious threat to people's health and life in China.At present， drug therapy has been primarily adopted clinically in the treatment of the liver injury.In-depth investigation of the mechanism of liver-protective drugs is of great significance to the prevention and treatment of clinical liver diseases.In recent years， with the development of the medical industry in China， an increasing number of studies have focused on the treatment of liver injury with Chinese medicine.Compared with western medicine， Chinese medicine is advantageous in few side effects and overall regulation， which plays a pivotal role in liver protection.However， its underlying mechanism in liver protection still needs to be further studied due to its complex compositions and diverse targets.Metabolomics， a new approach to studying the metabolic pathway of biological systems， provides integral and systematic views in the investigation of liver protection with Chinese medicine. By virtue of metabolomics， the mechanism of Chinese medicine in multi-target and multi-pathway liver protection can be analyzed comprehensively， and the corresponding biomarkers can also be screened out. The authors analyzed the studies of the treatment of chemical liver injury models induced by carbon tetrachloride （CCl4）， dimethylnitrosamine （DMN）， α-naphthyl isothiocyanate （ANIT）， and alcohol by Chinese medicinal compounds， single herbal medicines， and monomers of Chinese medicine based on metabolomics， and summarized the biomarkers and related metabolic pathways of Chinese medicine in the intervention of each type of liver injury， aiming at providing a reference for the further research and clinical application in the treatment of different types of liver injuries by Chinese medicine.
Keywords：metabolomics;Chinese medicine;liver protection;chemical liver injury;biomarker;mechanism of action
Abstract：The brain and heart has a tight relationship upon physiology and pathology during the development of cardiovascular and cerebrovascular diseases with clinical conditions interacting with each other and complex pathological mechanisms. Clinical studies prove that cerebral diseases such as stroke usually happen with cardiac diseases as complications， and cardiac diseases such as atrial fibrillation can also cause cerebral diseases， which can even aggravate brain atrophy and lead to cognitive impairment. Traditional Chinese medicine （TCM） also believes that “the substance of Shenming is located in the brain and the function in the heart.” Specifically， Yuanshen is in the brain and Shishen is in the heart， which thus makes the heart and brain closely related. If Shenming in any of them is impaired， the other one would also be injured. Therefore， the pathological mechanisms of brain-heart mutual damage have become one of the current research hotspots. This paper， combined with the clinical research status of the brain-heart mutual damage， summarized its pathological mechanisms from the perspectives of inflammatory responses， dysregulation of autonomic nervous system， apoptosis， energy metabolism and oxidative stress. The necessity of “brain-heart concurrent regulation” was proposed and the research progress on the treatment of cerebral and cardiac diseases with TCM represented by Naoxintong capsule was profiled in the light of heart and brain function described by TCM and the holistic concept of TCM treating cerebral and cardiac diseases. This paper reviews the pathological mechanisms of brain-heart mutual damage and the research progress on its treatment with TCM， which can provide reference for the prevention and treatment of cardiovascular and cerebrovascular diseases and further research on them.
Keywords：brain-heart mutual damage;pathological mechanisms;inflammatory responses;dysregulation of autonomic nervous system;prevention and treatment with traditional Chinese medicine
Abstract：Gout is the second largest metabolic disease in China， which can cause joint tissue damage and a variety of chronic diseases， and seriously affect human life and health. The increase in uric acid caused by disorder of purine metabolism or abnormal uric acid excretion is the biochemical basis of its pathogenesis. Western medical treatment mainly uses anti-inflammatory drugs such as colchicine， non-steroidal anti-inflammatory drugs， and uric acid lowering drugs such as febuxostat and benzbromarone， which have obvious effects， but there are problems such as easy to recurrence after drug withdrawal and more adverse reactions. Traditional Chinese medicine （TCM） has a long history in the treatment of gout， and has the advantages of multi-channel， multi-target， and multi-level symptomatic treatment. It exerts therapeutic effects through lowering uric acid， anti-inflammatory， anti-oxidation， and protecting the kidneys. Its curative effect is obvious and the adverse reaction rate is low. In recent years， there have been many studies on the mechanism of TCM for gout animal models. Based on the review of relevant literature in recent years， this article has systematically sorted out the pathogenesis of gout， the mechanism of TCM for gout and related experimental design. The paper summarized and analyzed the mechanism of TCM in the treatment of gout from the aspects of regulating the level of inflammatory factors， inhibiting oxidation reaction， reducing uric acid and regulating signaling pathway， so as to provide reference for the research and development of drugs for gout.
Keywords：gout;traditional Chinese medicine （TCM）;mechanism;pharmacodynamic evaluation;hyperuricemia;anti-inflammatory;anti-oxidant
Abstract：Ovarian cancer， one of gynecological malignancies， is often diagnosed at the late stage because of the atypical early symptoms and has become a major killer of women. Research has found that the co-evolution of tumor cells and their surrounding microenvironment is an important cause for the occurrence and development of ovarian cancer. It is believed in traditional Chinese medicine （TCM） that Yin and Yang are the roots of everything， and their balance， namely Yin being at peace and Yang being compact （“Yin Ping Yang Mi”） is a sign of good health. The mutual opposition， restriction， and rooting of Yin and Yang as well as their waning and waxing and transformation are the keys to maintaining the balance. In TCM， ovarian cancer falls into the category of abdominal mass， which results from the struggle between healthy Qi and evil Qi. When the healthy Qi is deficient and the evil Qi is excessive， the balance between Yin and Yang will be destroyed， triggering the body and ovarian cancer microenvironment as well as the relevant factors in the inflammatory microenvironment to be mutually opposed， restricted， and transformed， highly consistent with the dynamic development of Yin and Yang. At present， the studies concerning TCM intervention in the inflammatory microenvironment of ovarian cancer mostly focus on the signaling pathways to reveal the advantages of TCM multiple components against cancer cells via multiple targets， but they fail to explain the TCM efficacy from the perspective of Yin-Yang balance. Therefore， guided by the concept of Yin-Yang balance， this paper macroscopically and microscopically explored the effects of the changed factors in inflammatory microenvironment on the occurrence and development of ovarian cancer， and put forward that the prevention and control principles of ovarian cancer should lie in the "adjustment of Yin-Yang balance"， accompanied by healthy Qi reinforcement and pathogen elimination. This paper has laid the foundation for the elucidation of modern research achievements regarding the ovarian cancer microenvironment with TCM theory and provide a theoretical basis for the clinical treatment of ovarian cancer with integrated TCM and western medicine.
Abstract：As a typical representative of heat-clearing and detoxifying prescriptions， Huanglian Jiedutang （HJT） has various pharmacological activities and is widely used in clinical practice. The articles concerning the effect and clinical application of HJT published in recent years were retrieved from such databases as China National Knowledge Infrastructure （CNKI） and PubMed to figure out HJT efficacy， especially in anti-inflammation， the corresponding action pathways， and its clinical application. It was found that the anti-inflammatory effect mainly resulted from HJT regulation of multiple pathways including interleukin-17 （IL-17） signaling pathway， tumor necrosis factor （TNF） signaling pathway， Toll-like receptor 4 （TLR4） signaling pathway， and neutrophil chemotaxis. The inflammatory cytokines in the serum were reduced via these pathways and thus the inflammation was inhibited. Because of its unique anti-inflammatory advantage， HJT has been widely used for the treatment of cardiovascular and cerebrovascular diseases， digestive system diseases， skin inflammation， sepsis， and other infectious diseases. In view of this， the paper reviewed the anti-inflammatory effect and clinical application of HJT， aiming to provide a reference for further research.