Abstract：ObjectiveTo observe the effect of Qigesan on the proliferation and apoptosis of the human esophageal cancer cell EC9706， and the effect on miR-133a/protein kinase B（Akt）/mammalian target of rapamycin （mTOR） signaling pathway.MethodThe effective constituent of Qigesan was extracted by ethyl acetate. Thiazolyl blue tetrazolium bromide（MTT） colorimetric assay was used to determine the dosage of Qigesan on cells and to detect the effect of Qigesan on the proliferation of EC9706 cells. The effect of Qigesan on apoptosis of EC9706 cells was detected by flow cytometry. The effect of Qigesan on miR-133a and insulin-like growth factor 1 receptor（IGF-1R） mRNA expression was detected by Real-time quantitative polymerase chain reaction （Real-time PCR） . The protein expression of Akt and mTOR in EC9706 cells was detected by Western blot.ResultQigesan can inhibit the proliferation of EC9706 cells in a dose-dependent manner（P<0.01）. Inhibitory concentrations 30% inhibition concentration（IC30） 40 mg·L-1 and median inhibition concentration（IC50） 80 mg·L-1 were selected for follow-up experiments. Compared with the blank group， both the inhibitor group and the combination drug group can inhibit the proliferation of EC9706 cells （P<0.01）. The inhibitor at 0.25 μmol·L-1 was selected for subsequent experiments. Compared with the blank group， Qigesan 80 mg·L-1 dose group could significantly promote the late apoptosis rate and total apoptosis rate of EC9706 cells（P<0.05）， and the 40 mg·L-1 dose group could significantly promote the late apoptosis rate of EC9706 cells（P<0.05）， which shows synergistic effect after concomitant use with Akt/mTOR inhibitor（P<0.05）. Compared with the blank control group， each group can effectively increase expression of miR-133a（P<0.05）. The combination of inhibitor and traditional Chinese medicine（TCM） has obvious promotion effect. Compared with blank control group， the expressions of Akt and mTOR were significantly decreased in each group（P<0.05）. Compared with single medication， the expressions of Akt and mTOR were decreased in combination of inhibitor and TCM group.ConclusionQigesan can inhibit the growth of EC9706 cells and promote apoptosis， and its inhibitory mechanism may be related to the Akt/mTOR signaling pathway by regulating the expression of miR-133a.
Keywords：Qigesan;EC9706 cell;proliferation;apoptosis;miR-133a/protein kinase B（Akt）/mammalian target of rapamycin （mTOR） signaling pathway
Abstract：ObjectiveTo control the quality of the reference sample of Wenjingtang by establishing the specific chromatograms.MethodOn the basis of analyzing 15 batches of Wenjingtang freeze-dried powder samples， a high performance liquid chromatography （HPLC） specific chromatogram analysis method of Wenjingtang was established. The system adaptability was investigated and the retention time， relative retention value and deviation caused by different chromatographic columns and instruments were calculated by using the same brand of chromatographic columns， four different brands of chromatographic columns and instruments from three different manufacturers. The precision， repeatability and stability of this method was further completed. The possible chemical components of the freeze-dried powders were speculated and identified by ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF-MSn）. Chromatographic separation was performed on ACQUITY UPLC BEH C18 column （2.1 mm×100 mm， 1.7 μm） with acetonitrile （A）-0.1% formic acid aqueous solution （B） as mobile phase for gradient elution （0-2.8 min， 10%A； 2.8-8.0 min， 10%-18%A； 8.0-12.2 min， 18%-25%A； 12.2-15.3 min， 25%-40%A； 15.3-17.4 min， 40%A； 17.4-20.5 min， 40%-90%A）， and column temperature was set at 30 ℃ with flow rate of 0.4 mL·min-1. Mass spectrometry was performed on electrospray ionization， data were collected under positive and negative ion modes， and the detection range was m/z 50-1 600.ResultTen characteristic peaks were selected as the distinguishing features in this specific chromatograms， and eight of them were identified by comparing with the reference standards， including paeoniflorin （peak 1）， liquiritin apioside （peak 2）， liquiritin （peak 3）， ferulic acid （peak 4）， iquiritigenin （peak 6）， cinnamaldehyde （peak 8）， paeonol （peak 9）and glycyrrhizic acid （peak 10）. By mass spectrometry analysis， 30 compounds were identified， and the source of medicinal materials were assigned. It mainly contained triterpenoid saponins and flavonoids from Glycyrrhizae Radix et Rhizoma， ginsenosides from Ginseng Radix et Rhizoma， monoterpenoid glycosides and tannins from Paeoniae Radix Alba， steroids in Achyranthis Bidentatae Radix， phenolic acids in Angelicae Sinensis Radix.ConclusionThe established characteristic chromatographic analysis method of Wenjingtang is simple， stable and repeatable. The chemical composition of the freeze-dried powder of Wenjingtang is basically defined by mass spectrometry identification and source attribution， which can provide reference for the development and quality control of Wenjingtang in the future.
Keywords：famous classical formulas;Wenjingtang;reference sample;specific chromatogram;ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF-MSn）;relative retention time;quality control
Abstract：By systematically sorting out the ancient medical books and modern clinical literature of Yiguanjian， the historical evolution of this formula， including its source， composition， origin， processing， dosage， preparation and usage， functions and indications， evolution of prescription meaning， is textual so as to clarify the historical evolution and clinical application of Yiguanjian. On the basis of fully considering the actual demand of development of famous classical formula preparation and the usage habit of modern clinical practice， the feasible development suggestions were put forward. Yiguanjian is composed of six herbs， which is derived from Yifang Jiedu (《医方絜度》) . It is an ancient book of traditional Chinese medicine edited by QIAN Min-jie in Qing dynasty. The original medicinal plants and medicinal parts of the formula were basically the same as those recorded in the 2020 edition of Chinese Pharmacopoeia. The raw products should be selected for decoction pieces and processed according to the methods recorded in the 2020 edition of Chinese Pharmacopoeia. The reference dose of the medicine in this formula is set out in Yifang Jiedu. According to dosage of one Qian（钱）=3.73 g， the dosages of Glehniae Radix， Ophiopogonis Radix and Angelicae Sinensis Radix were 5.60 g， the dosages of Lycii Fructus and Rehmanniae Radix were 11.19 g， the dosage of Toosendan Fructus was 7.46 g. These decoction pieces were boiled and warm decoction was taken. According to ancient medical records， the formula always has the effect of nourishing Yin and relieving Qi of liver. It is used to treat syndrome of stagnation of liver-Qi and deficiency of liver-Yin and kidney-Yin， which can be seen with pain in chest， stomach and flank， acerbity and vomiting， dry throat and mouth， red tongue， weak pulse or deficiency of string and hernia. Here， the source， processing and others of Yiguanjian were clarified， providing a literature reference for the development and application of this famous classical formula.
Keywords：Yiguanjian;prescription textual research;origin;processing;famous classical formulas;functions and indications;clinical application
Abstract：ObjectiveTo explore the microecological mechanisms of Shenling Baizhusan （SLBZ） and Lizhongtang （LZ） in treating antibiotic-associated diarrhea （AAD） based on changes in the diversity of intestinal butyrate-producing bacteria.MethodSD rats were randomly divided into an SLBZ group （5.5 g·kg-1·d-1） and an LZ group （5.5 g·kg-1·d-1）. The gut microbiota disturbance model was induced by intragastric administration of clindamycin hydrochloride （315 mg·kg-1·d-1） and AAD model by Clostridium difficile. Subsequently， the rats were treated correspondingly. Fecal samples at different stages were collected and the total DNA was extracted. Polymerase chain reaction （PCR） amplification was performed with the primers of butyryl coenzyme A （CoA）-CoA transferase genes. The PCR products were cloned and sequenced to analyze the diversity response of butyrate-producing bacteria.ResultAfter treatment， both groups showed increased food uptake， formed feces， glossy and smooth fur， and improved activity and sensitivity. With the butyryl CoA-CoA transferase gene as the molecular marker， 297 sequences of butyrate-producing bacteria in the SLBZ group （SPD for short） and 300 sequences of butyrate-producing bacteria in the LZ group （LPD for short） were obtained. In the SLBZ group， 98， 100， and 99 sequences of SPD were obtained at the normal stage， the modeling stage， and the treatment stage， respectively， belonging to 8， 3， and 6 operational taxonomic units （OTUs）， with similarity ranges of 78%-97%， 86%-99% ， and 81%-97%. The number of OTUs recovered to 75% of the normal level after treatment. In the LZ group， 100 sequences of LPD were obtained at the normal stage， the modeling stage， and the treatment stage， respectively， belonging to 6， 2， and 4 OTUs， with similarity ranges of 83%-97%， 92%-99%， and 85%-99%. The number of OTUs recovered to 80% of the normal level after treatment. Butyrate-producing bacteria were present in all stages of the two groups， dominated by Firmicutes， accounting for more than 98% of the total number. The effects of SLBZ on SPD at the genus level were observed in the significant decrease in Clostridium abundance and the significant increase in Eubacterium abundance. The effect of LZ on LPD was mainly concentrated on the Roseburia at the genus level， and LZ also increased the abundance of Eubacterium， Lacrimispora， and Clostridium. According to the phylogenetic tree， the classification of butyrate-producing bacteria increased from five clusters to seven clusters after SLBZ treatment， while that increased from three clusters to nine clusters after LZ treatment.ConclusionIn the treatment of AAD， SLBZ and LZ can regulate the structure and abundance of butyrate-producing bacteria in the intestine， restore their diversity， and improve the instability of the intestinal microecological environment.
Abstract：ObjectiveTo observe and compare the protective effects of Tongqiao Huoxue decoction （TQHX） prepared by three methods against cerebral ischemia-reperfusion injury （CIRI）， and to explore its mechanism through the glutamate （Glu） metabolic pathway in astrocytes.MethodThe male SD rats of SPF grade were subjected to CIRI model induction by the modified middle cerebral artery occlusion method. The model rats were randomly divided into a model group， a sham operation group， and water-decocted， wine-decocted， and alcohol-extracted TQHX （6.3 g·kg-1·d-1） groups. The rats were treated correspondingly for 7 days. Those in the sham operation group and the model group were treated with an equal volume of normal saline by gavage. After the final treatment， the neurological function of rats was assessed by the modified neurological severity score （mNSS）. Hematoxylin-eosin （HE） staining was used to observe the morphological changes of ischemic brain tissues in rats. High-performance liquid chromatography （HPLC） was used to detect glutamate （Glu） in ischemic brain tissues. The expression of glutamate transporter-1 （GLT-1） and glial fibrillary acidic protein （GFAP） and co-expression of glutamine synthetase （GS） and GFAP in ischemic brain tissues were detected by immunofluorescence assay. Western blot was used to detect the protein expression of GFAP， GLT-1， and GS.ResultCompared with the sham operation group， the model group showed increased mNSS （P<0.01）， large necrosis of cerebral cortex in ischemic brain tissues with disordered cell arrangement， obscure boundary， intracellular edema， and inflammatory infiltration， elevated Glu in ischemic brain tissues （P<0.01）， declining GLT-1-GFAP co-expression and GS-GFAP co-expression （P<0.01）， up-regulated expression of GFAP protein， and reduced protein expression of GLT-1 and GS（P<0.05，P<0.01）. Compared with the model group， the TQHX groups showed decreased mNSS （P<0.01）， relieved injury in the cerebral cortex and hippocampal nerve cells in ischemic brain tissues， reduced Glu expression（P<0.05，P<0.01）， elevated co-expression of GLT-1 and GFAP （P<0.05，P<0.01）， and up-regulated protein expression of GFAP and GLT-1（P<0.05，P<0.01）. The co-expression of GS and GFAP （P<0.05，P<0.01）and the expression of GS （P<0.01）were increased in the wine-decocted and alcohol-extracted TQHX groups. Compared with the water-decocted TQHX group， the alcohol-extracted group showed increased GLT-1-GFAP and GS-GFAP co-expression（P<0.05）； the wine-decocted and alcohol-extracted TQHX groups exhibited elevated GS protein expression （P<0.05）； the alcohol-extracted TQHX group displayed declining Glu content （P<0.01） and increased protein expression of GFAP and GLT-1 （P<0.05， P<0.01）. Compared with the wine-decocted TQHX group， the alcohol-extracted TQHX group showed increased protein expression of GFAP and GLT-1（P<0.05，P<0.01）.ConclusionTQHX prepared by three methods can improve neurological deficits in CIRI rats. The effect is presumedly achieved by promoting the further activation of astrocytes， increasing the expression of GLT-1 and GS， promoting the clearance of Glu accumulated in the synaptic cleft by astrocytes through the Glu-glutamine （Gln） circulation， and reducing the excitotoxicity of Glu. The alcohol-extracted TQHX group was superior to the water-decocted and wine-decocted TQHX groups in reducing the content of Glu in ischemic brain tissues， promoting the activation of astrocytes， and enhancing the protein expression of GLT-1 and GS.
Abstract：ObjectiveTo investigate the effect of Shuangshen Ningxin capsules （SSNX） on cardiac hemodynamics and cardiac function in rats with coronary microvascular dysfunction.MethodRats were randomly divided into a sham operation group， a model group， a nicorandil group （5 mg·kg-1）， and high- （180 mg·kg-1）， medium- （90 mg·kg-1）， and low-dose （45 mg·kg-1） SSNX groups. Rats received corresponding drugs for 7 days. Two hours after the last administration， the model of coronary microvascular dysfunction was induced by left ventricular injection of embolic microspheres （40-120 μm， about 1 000 microspheres）. Twenty-four hours after modeling， left ventricular internal dimension in diastole （LVIDd）， left ventricular internal dimension in systole （LVIDs） left ventricular end-diastolic volume （LVEDV）， left ventricular end-systolic volume （LVESV）， stroke volume （SV）， cardiac output （CO）， left ventricular ejection fraction （EF）， and left ventricular shortening rate （FS） were detected by echocardiography. Cardiac catheterization was used to observe the arterial systolic blood pressure （SBP）， diastolic blood pressure （DBP）， left ventricular systolic pressure （LVSP）， left ventricular end-diastolic pressure （LVEDP）， maximum rate of increase in left ventricular pressure （LV+dp/dtmax）， and maximum rate of decrease in left ventricular pressure （LV-dp/dtmax）， and the mean arterial pressure （MAP） was calculated. Heart rate （HR） was calculated according to Ⅱ lead ECG. Biochemical analysis was carried out to detect the activities of creatine kinase （CK）， creatine kinase-MB （CK-MB）， and lactate dehydrogenase （LDH）. Enzyme-linked immunosorbent assay （ELISA） was used to detect serum cardiac troponin T （cTnT）. Western blot was used to detect the protein expression of Caspase-3， Bcl-2， and Bax， and 2，3，5-triphenyltetrazolium chloride （TTC） staining to observe the area of myocardial infarction. Hematoxylin-eosin （HE） staining was used to observe the morphological changes of the myocardium.ResultAs revealed by echocardiography， compared with the sham operation group， the model group showed reduced SV， CO， EF， and FS （P<0.01）， and increased LVIDs and LVEDV （P<0.01）. Compared with the model group， the SSNX groups showed increased EF （P<0.05， P<0.01） and FS （P<0.01）， and the high- and medium-dose SSNX groups displayed reduced LVIDs and LVESV， and increased LVEDV， SV， and CO （P<0.05， P<0.01）. SBP， DBP， MAP， LVSP， LV+dp/dtmax， and LV-dp/dtmax in the model group were lower than those in the sham operation group （P<0.01）， while there was no significant difference in HR. SSNX improved hemodynamics of rats， and increased SBP， DBP， MAP， LVSP， LV+dp/dtmax， LV-dp/dtmax， and HR as compared with the model group （P<0.05， P<0.01）. The serum CK， LDH， CK-MB， and cTnT levels in the model group were higher than those in the sham operation group （P<0.01）. Compared with the model group， SSNX groups reduced serum CK， LDH， CK-MB， and cTnT （P<0.05， P<0.01）. Compared with the sham operation group， the model group displayed increased expression of Caspase-3 protein in the myocardium （P<0.01） and reduced expression of Bcl-2 protein （P<0.05）. The expression of Caspase-3 protein in the myocardium of SSNX groups was lower than that in the model group， and statistical difference was observed between the low-dose SSNX group and the model group （P<0.05）. Compared with the model group， the SSNX groups exhibited increased expression of Bcl-2 in the rat myocardium， and the statistical difference was observed in the high-dose SSNX group （P<0.01）. As demonstrated by the TTC staining， compared with the model group， SSNX groups showed reduced areas of myocardial infarction （P<0.01）. The HE staining indicated that the pathological injury in myocardial tissues of the SSNX groups was relieved as compared with that in the model group.ConclusionSSNX can significantly enhance the cardiac function after coronary microvascular dysfunction caused by embolic microspheres， improve cardiac hemodynamics， reduce the area of myocardial infarction， and decrease CK， LDH， CK-MB， and cTnT levels. The mechanism may be related to the inhibition of cardiomyocyte apoptosis to protect the myocardium.
Keywords：Shuangshen Ningxin capsule;Qi-benefiting and blood-activating Chinese medicine;coronary microvascular dysfunction;cardiac function;hemodynamics
Abstract：ObjectiveTo establish a model of cervical spondylosis of vertebral artery type （CSA） in rats by mixed modeling method， and observe the intervention effect of Panlongqi tablet （PLQT） on CSA rats.MethodSD rats were divided into a normal control group， a model group， low- （0.16 g·kg-1）， medium- （0.32 g·kg-1）， and high-dose （0.64 g·kg-1） PLQT groups， and a Jingfukang granule （JFK， 1.35 g·kg-1） group. The rats were treated correspondingly 24 hours after modeling for eight weeks， and those in the normal control group received an equal volume of normal saline by gavage. The limb movement was tested by the inclined plate assay， vertebral artery flow volume by multi-mode high-frequency sound wave for small animals， and microcirculatory blood flow in the pia mater by the laser Doppler. The imaging of the cervical spine was recorded and scored by X-ray micro-computed tomography （Micro CT）. Enzyme-linked immunosorbent assay （ELISA） was used to detect serum levels of endothelin-1 （ET-1）， nitric oxide （NO）， tissue plasminogen activator （t-PA）， and plasminogen activator inhibitor （PAI）.ResultCompared with the normal control group， the model group showed decreased limb movement， vertebral artery flow volume， and microcirculatory blood flow in the pia mater， and increased imaging of the cervical spine and score （P<0.05，P<0.01）. PLQT could dose-dependently improve the motor function， increase the vertebral artery flow volume and microcirculatory blood flow in the pia mater， and reduce the degree and score of imaging of the cervical spine in CSA rats（P<0.05，P<0.01）. The serum levels of NO and t-PA were decreased and those of ET-1 and PAI were increased in the model group as compared with those in the normal control group， while such changes were reversed by PLQT treatment（P<0.05，P<0.01）.ConclusionPLQT can enhance the limb movement， promote the vertebral artery flow volume and microcirculatory blood flow in the pia mater， improve the degree of imaging of the cervical spine， regulate the vasomotor function， and improve the coagulation and fibrinolysis system of CSA rats， which shows good potential for the treatment of CSA.
Keywords：Panlongqi pill;rats;cervical spondylosis of vertebral artery type;vertebral artery blood flow;vasomotor function
Abstract：ObjectiveTo observe the intervention of phlegm-stasis co-treatment on the myocardial Toll-like receptor 4 （TLR4）/nuclear factor-κB （NF-κB）/nuclear factor-κB inhibitor （IκB） signaling pathway， and to investigate its mechanism in improving myocardial inflammation in rats with diabetes mellitus （DM）.MethodForty-five male SD rats of SPF grade were randomly divided into a normal group， a phlegm-resolving （Xiao Xianxiongtang， 4.05 g·kg-1） group， a stasis-resolving （Xuefu Zhuyutang， 7.02 g·kg-1） group， a co-treatment （Didang Xianxiong decoction， 8.10 g·kg-1） group， an alagebrium chloride （3 mg·kg-1） group， and a model group. Except for normal group， the other rats was induced by a single intraperitoneal injection of 55 mg·kg-1 streptozotocin （STZ） to establish DM model. After adaptive feeding for three weeks， the rats were treated correspondingly by gavage daily for eight weeks. Rats were sampled under anesthesia. Enzyme-linked immunosorbent assay（ELISA） was used to detect the protein expression of TLR4 and tumor necrosis factor-alpha （TNF-α） in myocardial tissues. The expression levels of NF-κB p65 and IκBα were detected by immunohistochemistry. NF-κB p65， IκBα， TNF-α， and TLR4 mRNA expression levels were detected by real-time fluorescence-based quantitative polymerase chain reaction（Real-time PCR）.ResultThe protein and mRNA levels of TLR4， NF-κB p65， IκBα， and TNF-α were higher in the model group than those in the normal group （P<0.01）. TLR4， NF-κB p65， IκBα， and TNF-α protein and mRNA expression levels were reduced to varying degrees in the groups with drug intervention as compared with those in the model group （P<0.01）. The inter-group comparison revealed that the co-treatment group showed more manifest reduction in protein and mRNA expression levels of TLR4， NF-κB p65， IκBα， and TNF-α than the phlegm-resolving group and the stasis-resolving group （P<0.05，P<0.01）.ConclusionThe co-treatment of phlegm and stasis can improve myocardial inflammation in DM rats， with superior effect to either the phlegm-resolving method or the stasis-resolving method. The underlying mechanism may be related to the inhibition of TLR4/NF-κB/IκB signaling pathway activation.
Keywords：diabetic cardiomyopathy;co-treatment of phlegm and stasis;Didang Xianxiongtang;Toll-like receptor 4 （TLR4）/nuclear factor-κB （NF-κB）/NF-κB inhibitor （IκB） signaling pathway;inflammatory response
Abstract：ObjectiveTo investigate the effect and mechanism of Yiqi Fuzheng Jiedu decoction （YQFZJDD） on autophagy and growth of A549 cells.MethodA549 cells were intervened with YQFZJDD-containing serum prepared in advance. The levels of microtubule-associated protein 1 light chain 3 （LC3）， homologue of yeast autophagy-related gene 6 （Beclin1）， p62， p53， adenosine 5'-monophosphate-activated protein kinase （AMPK）， phosphorylated AMPK （p-AMPK）， mammalian target of rapamycin （mTOR）， and phosphorylated mTOR （p-mTOR） were detected by Western blot assay， and microtubule-associated protein 1A/1B light chain 3B （MAP1LC3B） by immunofluorescence （IF） assay. The proliferation， invasion， and senescence of A549 cells were separately measured by 5-ethynyl-2'-deoxyuridine （EDU） staining， Transwell assay， and β-galactosidase staining. In the presence of autophagy inhibitor 3-methyladenine （3-MA， 5 mmol·L-1）， the cells were divided into the 10% fetal bovine serum （blank） group， 10% control serum （control） group， low-， medium-， and high-dose 10% YQFZJDD-containing serum groups， and high-dose 10% YQFZJDD-containing serum + 3-MA group， followed by the measurement of A549 cell proliferation， invasion， and senescence. In the adoption of p53 inhibitor Pifithrin-α （PFT-α， 10 μmol·L-1）， the cells were divided into the control group， PFT-α group， high-dose YQFZJDD-containing serum group， and high-dose YQFZJDD-containing serum + PFT-α group. Then the LC3-Ⅱ and Beclin1 expression， MAP1LC3B fluorescence intensity， as well as A549 cell proliferation， invasion and senescence were determined.ResultCompared with the blank group and control group， YQFZJDD-containing serum at the medium and high doses up-regulated the protein expression levels of LC3-Ⅱ and Beclin1 in A549 cells after 48-h intervention in a dose-dependent manner （P<0.01）. Besides， YQFZJDD-containing serum at the low-， medium-， and high-doses down-regulated p62 and p-mTOR/mTOR expression （P<0.05， P<0.01）， elevated p53 and p-AMPK/AMPK （P<0.01）， decreased the number of proliferative and invasive cells， increased the number of senescent cells （P<0.01）， and enhanced the IF intensity of MAP1LC3B， with the optimal effect observed in the high-dose YQFZJDD-containing serum group （P<0.01）. Compared with the high-dose YQFZJDD-containing serum group， the high-dose YQFZJDD-containing serum + 3-MA group and high-dose YQFZJDD-containing serum + PFT-α group exhibited significantly increased proliferative and invasive cells but decreased senescent cells （P<0.05， P<0.01）. Meanwhile， the IF intensity of MAP1LC3B in the high-dose YQFZJDD-containing serum + PFT-α group was weakened and the LC3-Ⅱ and Beclin1 protein expression levels declined （P<0.05， P<0.01）.ConclusionYQFZJDD promotes the autophagy of A549 cells through the p53/AMPK signaling pathway to inhibit their proliferation， invasion and migration but accelerate senescence， thus playing a crucial role in inhibiting the progression of lung cancer.
Abstract：ObjectiveTo explore the mechanism of gentiopicroside （GPS） in preventing acute liver injury induced by carbon tetrachloride （CCl4） in mice and its effect on the Toll-like receptor 4 （TLR4）/myeloid differentiation factor 88 （MyD88）/nuclear factor-κB （NF-κB） signaling pathway.MethodSixty mice were randomly divided into a normal control group， a model group， a silymarin group （150 mg·kg-1）， and high- （200 mg·kg-1）， medium- （100 mg·kg-1）， and low-dose （50 mg·kg-1） GPS groups， with 10 in each group. The mice in the groups with drug intervention were administered correspondingly by gavage at 10 mL·kg-1， and those in the normal control group and the model group receive an equal volume of distilled water， once per day. Ten days after administration， mice in the normal control group were subjected to the intraperitoneal injection of peanut oil （10 mL·kg-1） and those in other groups were injected with peanut oil （10 mL·kg-1） containing 0.12% CCl4 for the induction of acute liver injury model. After fasting for 16 hours， blood was collected from eyeballs and liver tissues were collected. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of liver tissues. The content or activities of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， total bilirubin （TBIL）， alkaline phosphatase （ALP）， total superoxide dismutase （T-SOD）， and γ-glutamyl transpeptidase （γ-GT） in the serum， malondialdehyde （MDA） and glutathione peroxidase （GSH-Px） in liver tissues were determined by biochemistry techniques. The levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6） in liver tissues were measured by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the protein expression of TLR4， MyD88， and NF-κB in liver tissues. The expression of phosphorylated NF-κB （p-NF-κB） was detected by immunohistochemistry.ResultCompared with the normal control group， the model group showed increased levels of ALT， AST， ALP， TBIL， γ-GT， and MDA （P<0.01）， and blunted activities of T-SOD and GSH-Px （P<0.01）. Compared with the model group， the high- and medium-dose GPS groups exhibited declining levels of ALT， AST， ALP， TBIL， γ-GT， and MDA （P<0.05， P<0.01） and potentiated T-SOD and GSH-Px activities （P<0.05， P<0.01）. Compared with the normal control group， the model group displayed elevated levels of TNF-α， IL-1β， and IL-6 in liver tissues （P<0.01） and increased protein expression of TLR4， MyD88， and p-NF-κB （P<0.01）. Compared with the model group， the high- and medium-dose GPS groups showed decreased TNF-α， IL-1β， and IL-6 content in liver tissues （P<0.05， P<0.01） and dwindled TLR4， MyD88， and p-NF-κB protein expression （P<0.05， P<0.01）.ConclusionGPS possesses a protective effect on mice with acute liver injury induced by CCl4， and its mechanism of action may be related to the regulation of TLR4/MyD88/NF-κB signaling pathway and inhibition of oxidative stress.
Abstract：ObjectiveTo investigate the effect of Elian granule on autophagy and the phosphatidylinositol -3 kinase （PI3K）/protein kinase B （PKB/Akt）/mammalian target of rapamycin （mTOR） signaling pathway in gastric tissue of rats with gastric cancer.MethodSPF SD rats were randomly divided into the normal， model， Elian granule， and Weifuchun groups. In addition to the routine feeding in the normal group， the model， Elian granule， and Weifuchun groups received N-methyl-N'-nitro-N-nitrosoguanidine （MNNG） to induce gastric cancer in rats， and they were respectively given normal saline， Elian granule aqueous solution （3.240 g·kg-1） and Weifuchun aqueous solution （0.390 g·kg-1） by gavage （ig） for 48 weeks. The gross changes of the stomach taken by laparotomy were observed by naked eyes. Hematoxylin-eosin （HE） staining was performed to observe the histopathological changes of the gastric tissue in rats. Real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot （WB） were used to detect the mRNA and protein expression of microtubule-associated protein 1 light chain 3 beta （LC3B）， Beclin1， p62， PI3K， Akt， mTOR in rat gastric tissue.ResultCompared with the normal group， the model group showed gastric distension， thinner gastric wall， pale gastric mucosa， atrophied and flat folds， disordered course， and visible nodules and vegetations. Compared with the model group， the Elian granule group demonstrated alleviated gastric distension， dark gastric mucosa， reduced folds， and regular course， with the thinned gastric wall improved and granular nodules observed occasionally. According to HE staining， compared with the normal group， the model group showed crowded and disordered rat gastric glands， diverse in shape， varied cell morphology， basophilic cytoplasm， large irregular hyperchromatic nuclei， visible mitosis， and infiltrated and destroyed muscularis mucosae. While compared with the model group， the arrangement of gastric glands was regular， and a few mildly atypical cells could be observed in rats of the Elian granule group. Compared with the normal group， the model group exhibited decreased expression of LC3B and Beclin1 mRNA and protein in gastric tissue （P<0.05）， and increased expression of PI3K， p62， Akt， and mTOR mRNA and protein （P<0.05）. Compared with the model group， the Elian granule group showed increased expression of LC3B and Beclin1 mRNA and protein in gastric tissue （P<0.05）， and decreased expression of PI3K mRNA and p62， Akt， and mTOR mRNA and protein （P<0.05）.ConclusionElian granule can improve the cell atypia of gastric tissue in rats with gastric cancer， and the mechanism may be related to the inhibition of the PI3K/Akt/mTOR signaling pathway to promote autophagy.
Keywords：Elian granule;gastric cancer;autophagy;phosphatidylinositol -3 kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway
Abstract：ObjectiveTo study the protective effect of polysaccharides from Plantaginis Semen （PSP） against renal injury in rats with membranous nephropathy （MN） and its influence on the gut microbiota to provide a theoretical basis for the further investigation of PSP in the treatment of MN.MethodThe MN model was induced by tail vein injection of cationic bovine serum albumin （C-BSA， 3.5 g·L-1） in rats with a modeling period of seven weeks. At the 4th week of modeling， the model rats were divided into a model group， a positive drug group （benazepril hydrochloride， 10 mg·kg-1·d-1）， a PSP high-dose group （PSP-H， 800 mg·kg-1·d-1）， a PSP medium-dose group （PSP-M， 400 mg·kg-1·d-1）， and a PSP low-dose group （PSP-L， 200 mg·kg-1·d-1） according to the random number table， with 10 in each group. Ten healthy rats were assigned to the normal control group. The rats in the normal control group and the control group received an equal amount of physiological saline by gavage， and those in the groups with drug intervention were administered correspondingly，once a day，for consecutive four weeks. The pathological changes of rat kidney and colon tissues were observed by optical microscopy. The enzyme-linked immunosorbent assay （ELISA） was used to detect the content of tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β） in the serum and colon tissues. The immunohistochemistry （IHC） was used to detect the protein expression of TNF-α and IL-1β in renal tissues. The 16S rRNA sequencing method was used to investigate the effect of PSP on the gut microbiota in MN rats.ResultCompared with the normal control group， the model group showed enlarged glomeruli， thickened basement membrane， atrophied colonic gland， increased TNF-α and IL-1β in the serum and colon tissues （P<0.05）， and elevated protein expression of TNF-α and IL-1β （P<0.01）. Compared with the model group， the positive drug group and the PSP-H group displayed shrunk glomerular capsules， relieved basement membrane thickening， and neatly arranged colonic mucosa in colon tissues， while the PSP-M and PSP-L groups were inferior in improving renal tissues and colon tissues. Additionally， the PSP-H and PSP-M groups showed declining TNF-α and IL-1β in the serum and colon tissues （P<0.05） and dwindled protein expression of TNF-α and IL-1β in the renal tissues （P<0.01）. No significant difference was observed in the PSP-L group. Compared with the normal control group， the model group showed increased abundance of Firmicutes and decreased abundance of Bacteroidetes. After PSP intervention， the abundance of Firmicutes was decreased， while that of Bacteroidetes was increased， and such changes were predominant in the PSP-H group.ConclusionPSP can effectively alleviate renal injury， reduce the expression of inflammatory factors， regulate the structure of gut microbiota， and improve the damaged intestinal barrier of MN rats.
Abstract：ObjectiveTo observe the clinical effect of compound Guizhencao granule （CGG） on grade 1 hypertension patients with dampness heat and blood stasis syndrome and its influence on related biological indicators and safety indexes.MethodA randomized，double-blind，placebo-controlled clinical trial design was used. 80 subjects who met the inclusion criteria were randomly divided into the treatment group （40 cases） and control group （40 cases）. On the basis of health education，patients in the treatment group received 6.5 g CGG，twice daily，for four weeks. Patients in the control group received CGG simulant in a similar dosing scheme. The 24-hour ambulatory blood pressure monitoring（24 h ABPM），traditional Chinese medcine（TCM） syndrome score，angiotensin Ⅱ （AngⅡ），endothelin-1 （ET-1），homocysteine（Hcy） and safety indexes were observed.ResultCompared with that before treatment， systolic blood pressure （SBP） and diastolic blood pressure （DBP） of the consulting room in the treatment group were significantly lower （P<0.01）， and there was no significant difference in the control group；The daytime DBP and 24 h DBP in the control group decreased significantly （P<0.01）， and the 24 h SBP， 24 h DBP， daytime SBP， daytime DBP， nighttime SBP and nighttime DBP in the treatment group decreased significantly （P<0.01）. The total effective rate of 24 h ABPM on the nighttime blood pressure was 57.14% （20/35） in the treatment group， which was significantly higher than 28.57% （10/35） in the control group （Z=-2.310， P<0.05）； The total effective rate of daytime blood pressure and 24-hour blood pressure treatment group increased， but the difference was not statistically significant. The TCM syndrome score of two groups was significantly lower than that of the control group before treatment （P< 0.05， P< 0.01）， and that of the treatment group was significantly lower than that of the control group after treatment （P<0.01）. The total effective rate of TCM syndrome score in the treatment group was 51.43% （18/35）， which was significantly higher than 28.57% （10/35） in the control group （χ2= 9.973， P<0.05）. Compared with that before treatment， the levels of ET-1 and Hcy in the control group decreased significantly （P<0.05）， and the levels of Ang Ⅱ， ET-1 and Hcy in the treatment group decreased significantly （P<0.01）； Compared with the control group after treatment， the levels of Ang Ⅱ and ET-1 in the treatment group decreased significantly （P < 0.01）.ConclusionCGG is safe and effective in reducing the blood pressure level，improving the TCM syndrome score，and regulating related biological indicators of patients with Grade 1 hypertension.
Abstract：ObjectiveTo explore the effect of Shenqi Maiwei Dihuangtang （SQMWDH） combined with acarbose on the level of 2-hour oral glucose tolerance test （2 h OGTT），body mass index （BMI）， and abdominal fat thickness in patients with impaired glucose tolerance （IGT）.MethodA total of 130 patients with IGT admitted to the First People's Hospital of Shuangliu District from February 2017 to January 2019 were divided into a control group and a treatment group by a random number table. All patients received conventional treatment， such as diet regulation and exercise. The patients in the control group received additional oral administration of acarbose，while those in the treatment group were treated with SQMWDH based on the control group. Fasting blood glucose （FBG），2 h OGTT， and glycated hemoglobin A1c（HbA1c）levels were measured by the blood glucose meter. Abdominal fat thickness was measured by ultrasound tomography，and serum total cholesterol （TC），triglyceride （TG），adiponectin， and leptin levels in fasting venous blood were measured.ResultAfter treatment，the total response rate of the treatment group was higher than that of the control group （95.00% vs. 81.67%， χ2=5.175，P<0.05）. Before treatment，there was no significant difference in FBG，2 h OGTT，HbA1c， BMI，waist circumference，abdominal fat thickness，TC，TG，adiponectin， and leptin of IGT patients between the two groups. After treatment，the levels of FBG，2 h OGTT，HbA1c， BMI，waist circumference，abdominal fat thickness，TC，TG，and leptin of IGT patients were lower than those before treatment in both groups （P<0.05）， and the treatment group was inferior to the control group（P<0.05，P<0.01）. The level of adiponectin after treatment was higher than that before treatment in both groups （P<0.05），and the treatment group was superior to the control group （P<0.05）.ConclusionSQMWDH combined with acarbose is effective in treating IGT patients by effectively reducing 2 h OGTT and abdominal fat thickness to alleviate obesity and improve the constitution of patients.
Keywords：Shenqi Maiwei Dihuangtang;acarbose;impaired glucose tolerance;oral glucose tolerance test （OGTT）;body mass index;abdominal fat thickness
Abstract：ObjectiveTo explore the clinical efficacy and safety of modified Heweitang in the treatment of functional dyspepsia （FD） due to liver-stomach disharmony and its regulation of gastrointestinal hormones and brain-gut peptides.MethodOne hundred and twenty-six eligible patients were randomized into a control group （62 cases） and an observation group （64 cases）. Patients in the observation group took the modified Heweitang granules with warm water 30 min after meals， 10 g/time， 3 times/day， while those in the control group took the corresponding placebo granules at the same dose in the same manner. The treatment in both groups lasted for four weeks. Before and after treatment， the four main symptoms including postprandial satiety， early satiety， upper abdominal pain， and upper abdominal burning sensation were scored， followed by the examination of gastric emptying （GE） and the scoring of the functional digestive disorders quality of life questionnaire （FDDQL）， 7-point global overall symptom scale （GOSS）， and liver-stomach disharmony syndrome. The cholecystokinin （CCK）， motilin （MTL）， gastrin （GAS）， serotonin （5-HT）， vasoactive intestinal peptide （VIP）， and substance P （SP） levels before and after treatment were detected， and then the safety was evaluated.ResultAfter treatment， the scores of the four main symptoms， GOSS， and liver-stomach disharmony syndrome in the observation group were lower than those in the control group （P<0.01）， while the GE rate and FDDQL scores in the observation group were higher （P<0.01）. The CCK， GAS， 5-HT， and VIP levels of the observation group declined as compared with those of the control group （P<0.01）， whereas the MTL and SP levels were elevated （P<0.01）. After treatment， the overall response rate in the observation group was （51/57）89.47%， higher than （15/56）26.79% in the control group （χ2=45.696， P<0.01）. No drug-related adverse reactions were found during the trial.ConclusionThe modified Heweitang is efficient and safe in relieving the main and related symptoms and traditional Chinese medicine （TCM） syndrome， regulating the secretion of gastrointestinal hormones and brain-gut peptides， promoting GE rate， and improving the quality of life of patients with FD due to liver-stomach disharmony.
Abstract：ObjectiveTo observe the clinical efficacy of modified Qiaohetang and Xiehuangsan in the treatment of acne due to dampness-heat accumulation and its influence on the levels of inflammatory factors and sex hormones.MethodOne hundred and sixty-eight eligible patients were divided into an observation group （84 cases） and a control group （84 cases） according to the random number table. Adapalene gel was applied externally in both groups， one time per day. In the control group， Jinhua Xiaocuo pills was taken orally， 4 g per time， three times a day. In the observation group， the modified Qiaohetang and Xiehuangsan was provided for oral administration， one bag per day. The treatment lasted for eight weeks. The Global Acne Grading System （GAGS） score， skin lesion count， dampness-heat accumulation syndrome score， and Dermatology Life Quality Index （DLQI） score were recorded before and after treatment， followed by the detection of interleukin-8 （IL-8）， IL-10， IL-17， interferon-γ （IFN-γ）， free testosterone （FT）， estradiol （E2） and sex hormone binding globulin （SHBG） before and after treatment as well as the safety evaluation.ResultThe GAGS， dampness-heat accumulation syndrome， and DLQI scores of the observation group were lower than those of the control group （P<0.01）. The counts of inflammatory skin lesions （papule and pustule）， non-inflammatory skin lesions， and total skin lesions in the observation group declined in contrast to those in the control group （P<0.01）. The IL-8， IL-17， IFN-γ and FT levels of the observation group were decreased as compared with those of the control group （P<0.05， P<0.01）， while the IL-10， E2， and SHBG levels were increased （P<0.01）. The overall response rate in clinical symptom alleviation of the observation group was 93.67%（74/79）， which was higher than 81.82%（63/77） of the control group （χ2=5.121， P<0.05）. The overall response rate in dampness-heat accumulation syndrome relief of the observation group was 92.41% （73/79）， still higher than 79.22% （61/77） of the control group （χ2=5.595， P<0.05）. No adverse reactions occurred after the oral administration of Chinese medicinal preparations.ConclusionThe modified Qiaohetang and Xiehuangsan combined with adapalene gel can reduce the skin lesion count and severity， relieve both clinical symptoms and dampness-heat accumulation syndrome， regulate the inflammatory response and sex hormones， and improve the quality of life of patients with acne of dampness-heat accumulation syndrome without inducing side effects.
Abstract：ObjectiveTo observe the clinical efficacy of Jiangshi Xuantong decoction （JSXT） in the treatment of peripheral neuropathy patients with arthralgia syndrome caused by wind， heat， and dampness，investigate the changes in common clinical evaluation indexes and electrophysiology，and verify its effectiveness and safety.MethodA total of 120 patients were randomly divided into a JSXT group （n=60） and a mecobalamin group （n=60）. The changes in Toronto clinical neuropathy scores （TCNSs），traditional Chinese medicine （TCM） syndrome scores，efficacy for TCM syndrome，electrophysiological results in the electromyogram， and safety indexes before and after JSXT treatment of peripheral neuropathy were observed to determine the clinical efficacy and safety of JSXT for peripheral neuropathy.ResultNo significant differences in various indexes among patients were observed before treatment. The TCM syndrome score，TCNS， and electrophysiological results in the electromyogram of the patients after treatment were significantly higher than those before treatment （P<0.05）. The evaluation indexes of the treatment group were significantly higher than those of the control group （P<0.05）. During the follow-up，the evaluation indexes in the treatment group increased with time，and the corresponding growth was higher than that in the control group. There were no significant adverse reactions in both groups，and the difference was not statistically significant.ConclusionJSXT in the treatment of patients with peripheral neuropathy is superior to the control in terms of clinical symptoms，relevant scores， and electrophysiological results，with definite clinical efficacy. It is proved safe and reliable and can effectively and quickly enhance peripheral neurological function，improve the quality of life，and alleviate the negative emotions of patients. Therefore，JSXT possesses a good clinical therapeutic efficacy on patients with peripheral neuropathy and is expected to provide a new treatment method for patients with peripheral neuropathy，which is worthy of further clinical promotion.
Keywords：Jiangshi Xuantong decoction;peripheral neuropathy;clinical research
Abstract：ObjectiveTo investigate the effects of polyethylene glycol 400 （PEG400） on the pharmacokinetics and anti-inflammatory effect of baicalin， and to preliminarily explore the anti-inflammatory effects of baicalin and its main metabolite baicalein 6-O-β-D-glucuronide （B6G） by molecular docking.MethodRats were randomly divided into two groups with water and PEG400 as the dissolving matrix， and rats were administrated the equal dose of baicalin aqueous solution （baicalin+water group） and baicalin PEG400 solution （baicalin+PEG400 group）. After the plasma samples were processed at different time periods， the concentrations of baicalin and B6G in rat plasma were determined by ultra performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS）， and pharmacokinetic parameters were processed by DAS 3.2.2 software. Mice were randomly divided into a blank group （normal saline， 20 mL·kg-1）， aspirin group （dose of 0.2 g·kg-1）， baicalin/baicalin+PEG400 high and low dose （3.0， 1.5 g·kg-1） groups， after continuous administration for 7 days， the mouse ear swelling and foot swelling models were established， and the swelling degree and swelling inhibition rate were calculated.ResultThe pharmacokinetic study showed that compared with baicalin+water group， the plasma concentrations of baicalin and B6G increased after administration of baicalin PEG400 solution， and the area under the curve （AUC0-t） increased by 2.36， 1.97 times， and the peak concentration （Cmax） increased by 2.12， 1.65 times， respectively. The results of mouse ear and foot swelling inflammation models showed that the anti-inflammatory effect was enhanced after intragastric administration of baicalin PEG400 solution. In addition， molecular docking results showed that baicalin and B6G could site bind to multiple target proteins ［tumor necrosis factor （TNF）-α， interleukin （IL）-6， IL-1β， prostaglandin E2 （PGE2） and nuclear transcription factor-κB （NF-κB）］ with higher affinity， which was superior to the positive drug aspirin.ConclusionPEG400 can increase the plasma concentration of baicalin and its main metabolite B6G， and enhance the anti-inflammatory effect. Baicalin and B6G can form strong hydrogen bonds with various inflammatory factors and of nuclear transcription factors， it is speculated that baicalin and B6G jointly play an anti-inflammatory role.
Abstract：ObjectiveTo observe the effect of Yangxin Tongmaifang （YXTM） on endogenous metabolites in the myocardial tissue of rats with coronary heart disease due to blood stasis based on the metabolomics approach， and to explore its mechanism in the treatment of heart blood stasis syndrome.MethodA rat model of chronic myocardial ischemia due to heart blood stasis was established via the high-fat diet combined with intragastric administration of vitamin D3 and subcutaneous injection of isoproterenol （ISO）， followed by the intervention with YXTM. The metabolites in the myocardial tissues of rats in the normal group （n=8）， model group （n=8）， and YXTM group （n=8） were detected by ultra-high performance liquid chromatography coupled to high resolution mass spectrometry. The high-throughput metabolomics data were then subjected to multivariate statistical analysis using SIMCA 14.1， and the related metabolic pathways were analyzed with MetaboAnalyst.ResultThe myocardial sample points of rats in the three groups were located in different areas of the elliptical confidence interval. The normal group and the model group were completely separated. There existed some crossovers and overlaps between the YXTM group and the normal group. The heart blood stasis syndrome model was proved successfully replicated from the perspective of metabolic profiling， and YXTM had the potential to promote the body to return to a normal state. After the intervention with YXTM， six differential metabolites changed significantly. Such metabolic pathways as valine， leucine， and isoleucine biosynthesis， pantothenate and coenzyme A （CoA） biosynthesis， valine， leucine， and isoleucine degradation， biosynthesis of aminoacyl-transfer RNA synthetases， and purine metabolism were involved.ConclusionThe therapeutic effect of YXTM on heart blood stasis syndrome in rats is related to the improved levels of myocardial endogenous metabolites， and its mechanism involves phospholipid metabolism， amino acid metabolism， energy metabolism， inflammatory response， and platelet activation and aggregation.
Abstract：ObjectiveTo establish the ultra-performance liquid chromatography （UPLC） fingerprints of Artemisiae Argyi Folium and Artemisiae Argyi Folium processed with four excipients， and quantitatively analyze the 8 phenolic acids and flavonoids contained in them， in order to explore the quality evaluation method of Artemisiae Argyi Folium processed with four excipients.MethodUPLC was used with Shim-pack XR-ODS C18 column （2.0 mm×75 mm， 2.2 µm）， mobile phase of acetonitrile （A） -0.2% formic acid aqueous solution （B） for gradient elution （0-1 min， 10%A； 1-2 min， 10%-15%A； 2-17 min， 15%-18%A； 17-24 min， 18%-28%A； 24-36 min， 28%-38%A； 36-41 min， 38%-60%A； 41-45 min， 60%-100%A）， detection wavelength of 330 nm and flow rate of 0.2 mL·min-1. The UPLC fingerprints of Artemisiae Argyi Folium before and after processing were established， and analyzed by chemometrics. Contents of 5-caffeoylquinic acid， 3-caffeoylquinic acid， 4-caffeoylquinic acid， 3，4-dicaffeoylquinic acid， 3，5-dicaffeoylquinic acid， 4，5-dicaffeoylquinic acid， jaceosidin and epuatilin in the decoction pieces were determined.ResultThe fingerprints of Artemisiae Argyi Folium before and after processing were established， and the UPLC characteristic chromatograms of Artemisiae Argyi Folium before and after processing had good consistency， and the similarity was >0.94. Compared with Artemisiae Argyi Folium， the contents of 3-caffeoylquinic acid and 3，4-dicaffeoylquinic acid had no significant change after processing， the contents of jaceosidin and epuatilin decreased after processing， while the contents of 5-caffeoylquinic acid， 4-caffeoylquinic acid and 4，5-dicaffeoylquinic acid increased significantly （P<0.01）， their average increasing rates were 32.50%， 66.83%， 29.39%， respectively. And content of 3，5-dicaffeoylquinic acid was significantly decreased （P<0.01） ， and the average reduction rate was 51.25%.ConclusionThe contents of chemical components in Artemisiae Argyi Folium and Artemisiae Argyi Folium processed with four excipients have changed to a certain extent. Among them， 5-caffeoylquinic acid， 3，5-dicaffeoylquinic acid， 4-caffeoylquinic acid and 4，5-dicaffeoylquinic acid can be used as the key indicators for quality evaluation of Artemisiae Argyi Folium before and after processing.
Keywords：Artemisiae Argyi Folium processed with four excipients;fingerprint;Chinese medicine processing;ultra-performance liquid chromatography （UPLC）;determination;phenolic acids;flavonoids
Abstract：Illustrated Classic of Materia Medica （Ben Cao Tu Jing）， compiled in 1061， is regarded as the crystallization of the second national survey of Chinese medicinal resources in the history of China after the Newly Revised Materia Medica （Xin Xiu Ben Cao）， which serves as a precious source for exploring the Chinese medicinal resources in the Song Dynasty. The Illustrated Classic of Materia Medica （Ben Cao Tu Jing） covers 50 illustrations for Chinese medicinal materials named after geographical names of Jiangsu Province， 39 of which have been verified in today's Jiangsu Province. To be specific， Chinese medicinal materials in 32 illustrations can be traced back to their species， those in four illustrations to genera， and those in three illustrations to family. The remaining 12 illustrations remain to be further verified. The origins of most Chinese medicinal materials in the illustrations can be traced， which is attributed to their exquisiteness and lifelikeness as well as the detailed descriptions of the characters of the original plants and animals. In the textual research of these illustrations， we have noticed that there are inconsistencies between pictures and texts， and different illustrations may be derived from the same origin. This is believed to be related to SU Song's principle of collation that "for all Chinese medicinal materials presented as examples， if their shapes and categories recorded are not consistent with those in the literature， they will be retained. If there is any connection with the literature， they will be annotated according to literature to make their origins clear". As revealed by the distribution of Chinese medical materials in Jiangsu Province reported in the Illustrated Classic of Materia Medica （Ben Cao Tu Jing）， the utilization and development of Jiangsu medicinal materials during the Northern Song Dynasty were mainly concentrated in the low-altitude valleys and coastal areas. It could be seen from the illustrations of Chinese medicinal materials named after geographical names of today's Jiangsu Province that the production areas with a large number of illustrations were relatively developed in politics， economy， and transportation at that time， which enabled the medicinal resources to be fully utilized.
Keywords：Illustrated Classic of Materia Medica （Ben Cao Tu Jing）;Jiangsu Province;illustrations of medicinal materials;textual research;Chinese medicinal resources
Abstract：ObjectiveA strong antithrombotic protein component， named PvQ， was purified and enriched from total protein of Pheretima vulgaris， a traditional Chinese medicine. Moreover， we evaluated its fibrinolytic and anticoagulant activity， and expected to provide reference for the research on antithrombotic substances of Pheretima.MethodA rapid in vitro activity-oriented separation combined with the AKTA-Pure protein purification system conducted on P. vulgaris. Meanwhile， the fibrinolytic and anticoagulant activities of PvQ were measured by fibrin plate method and fibrinogen-thrombin time （Fibg-TT） method. And the in vitro thrombolysis assay was used for evaluating the lysis ability of PvQ to thrombus. Then the stability of PvQ was also analyzed for its anticoagulant activity at different pH and temperature.ResultThe PvQ was successfully enriched and its activity was determined to have significant fibrinolytic and anticoagulant activities. And the result of in vitro thrombolysis assay revealed that PvQ could hydrolyze more than 80% of thrombus after 5 h of incubation at 37 ℃. In addition， the changes of temperature and pH had significant effects on antithrombotic activity， and this study showed that PvQ was rapidly inactivated at ≥60 ℃ or in acidic conditions （pH<7）. While， the activity of PvQ was unaffected or less affected at ≤50 ℃ and under alkaline conditions.ConclusionA feasible preparation method of PvQ is established， and it can affect fibrin and fibrinogen at the same time， thus exerting a dual fibrinolytic effect and possessing significant fibrinolytic and anticoagulant activities. It provides a scientific interpretation for the treatment of thrombotic diseases by PvQ and a reference for the development of antithrombotic protein products of Pheretima.
Abstract：In this paper， through the collection and collation of ancient materia medica， medical books and medical formulary， combining with modern literature， the historical changes of the name， origin， position， harvesting time， medicinal parts， toxicity， functions and indications， processing methods of Rhododendri Mollis Flos （RMF） were systematically combed and verified， so as to provide reference for clinical application， processing standard and basic research of RMF. According to textual research， RMF is the dried flower of Rhododendron molle. In each historical period， there are many aliases and local names， being with phenomenon of homonyms and synonyms. RMF is mostly wild and planted in a small amount， harvesting time is mostly in March to April. However， the harvesting flowering period is differently described as initial bloom， full bloom and extensive bloom. RMF was first recorded in Shennong Bencaojing (《神农本草经》)， but it did not mention its medicinal parts. Then the flowers， fruits， roots are be used as medicine， but flowers are still the main medicinal parts. RMF had a long processing history， included fried， vinegar-fried， wine-fried， steamed， wine-steamed， vinegar-steamed， and many other processing methods in ancient times. However， at present， only raw products are used in clinical practice， and only a few modern books retain the methods of stir-fried and wine-steamed， believing that the processing can reduce toxicity of RMF.
Keywords：Rhododendri Mollis Flos;Rhododendron molle;herbal textual research;historical evolution on processing;visual analysis;traditional Chinese medicine
Abstract：ObjectiveTo analyze the compatibility rules of prescriptions containing Forsythiae Fructus based on data mining and explore the anti-inflammatory mechanism of Forsythiae Fructus based on network pharmacology，so as to provide reference for the rational clinical application of Forsythiae Fructus and the development of health foods and new Chinese medicines.MethodThe prescriptions containing Forsythiae Fructus in the Dictionary of Traditional Chinese Medicine Prescriptions were collected，based on which a clinical prescription database was constructed. The Chinese herbs combined with Forsythiae Fructus and the corresponding indications were subjected to frequency statistics，association rule analysis，and complex network analysis using SPSS Statistics 26，IBM SPSS Modeler 18.0，and Gephi 9.2. The active components and targets of Forsythiae Fructus for anti-inflammation were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP），BATMAN-TCM，and SEA，and the targets related to anti-inflammation from GeneCards，Online Mendelian Inheritance in Man （OMIM），CTD，and GenCLiP3. Following the analysis of protein-protein interactions （PPI） with STRING，a PPI network was constructed. The enrichment analysis was performed using Metascape，and the active component-anti-inflammation target-signaling pathway network of Forsythiae Fructus was constructed by Cytoscape 3.8.2.ResultAccording to the inclusion and exclusion criteria，2 245 prescriptions containing Forsythiae Fructus were harvested，involving 512 Chinese herbs，with a total usage frequency of 27 314. The Chinese herbs that were most frequently combined with Forsythiae Fructus （>800 times） were Glycyrrhizae Radix et Rhizoma （1 483 times），Scutellariae Radix （964 times），and Angelicae Sinensis Radix （842 times）. Hence，the herbal pairs "Forsythiae Fructus-Scutellariae Radix" and "Forsythiae Fructus-Angelicae Sinensis Radix" were further explored. The prescriptions containing Forsythiae Fructus could be utilized for the treatment of 29 kinds of diseases，and three representative disease categories including "carbuncle，gangrene，sores and ulcers"，"ophthalmic diseases and syndromes" and "epidemic diseases" are selected for data mining. There were 19 association rules obtained with "Forsythiae Fructus-Glycyrrhizae Radix et Rhizoma-Lonicerae Japonicae Flos-Angelicae Sinensis Radix" as the core herb combination for "carbuncle，gangrene，sores and ulcers". The clustering analysis revealed one multi-herb clustering group，four herbal pairs，and single herb Lonicerae Japonicae Flos，the complex network analysis four herbal modules，and the factor analysis six common factors. There were 23 association rules obtained with "Forsythiae Fructus-Glycyrrhizae Radix et Rhizoma-Scutellariae Radix-Angelicae Sinensis Radix" as the core herb combination for "ophthalmic diseases and syndromes". The clustering analysis revealed two multi-herb clustering groups and four herbal pairs，the complex network analysis four herbal modules，and the factor analysis five common factors. There were 28 association rules obtained with "Forsythiae Fructus-Glycyrrhizae Radix et Rhizoma-Menthae Haplocalycis Herba-Lonicerae Japonicae Flos" as the core herb combination for "epidemic diseases". The clustering analysis revealed three multi-herb clustering groups，one herbal pair，and two single herbs Forsythiae Fructus and Glycyrrhizae Radix et Rhizoma，the complex network analysis four herbal modules，and the factor analysis five common factors. As demonstrated by network pharmacology-based analysis，the core anti-inflammation components of Forsythiae Fructus were quercetin，luteolin，and kaempferol，and the core targets were phosphoinositide-3-kinase regulatory subunit 1 （PIK3R1），protein kinase B 1 （Akt1），and epidermal growth factor receptor（EGFR）. The biological pathways were mainly concentrated in proteoglycans in cancer，pathways in cancer，and PI3K/Akt signaling pathway，with such functions as inhibition of transcription factors，regulation of enzyme activity，and inflammation-related gene expression involved.ConclusionThis study employed a variety of data mining techniques to objectively，intuitively，and scientifically uncover the compatibility rules of Forsythiae Fructus in the treatment of high-frequency diseases. It has been found that Forsythiae Fructus is often combined with heat-clearing herbs，tonifying herbs，exterior-releasing herbs，and blood-activating and stasis-resolving herbs for diverse diseases and syndromes. Under the premise of clearing heat and removing toxin，reinforcing healthy Qi and dredging stagnation are also emphasized. According to the degree of internal heat exuberance，the heat-clearing herbs with different merits are combined. This study has revealed the unique advantages of Forsythiae Fructus in the treatment of specific diseases and syndromes as well as its multi-component，multi-target，and multi-pathway mechanisms in anti-inflammation，breaking through the limitations in modern clinical and experimental research of Forsythiae Fructus. These findings are of great significance for guiding the rational clinical application of Forsythiae Fructus and the development of health foods and new Chinese medicines，thus better accelerating the development of Chinese medicine health industry.
Keywords：Forsythiae Fructus;Dictionary of Traditional Chinese Medicine Prescriptions;data mining;network pharmacology;compatibility rules;anti-inflammation mechanism
Abstract：ObjectiveTo explore the regulatory effect of Huadu Sanyinfang on phosphatidylinositol 3 kinase（PI3K）/protein kinase B（Akt）/transcription factor nuclear factor-κB （NF-κB） in triple-negative breast cancer （TNBC） patients with qi-deficiency constitution based on the differential expression of miRNA.MethodBased on previous research results， this study conducted the bioinformatics analysis to predict the target genes responsible for regulating the differential expression of miRNA between patients with qi-deficiency constitution and those with moderate constitution， which were intersected with TNBC target genes. The resulting intersection targets were then subjected to Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis and protein-protein interaction （PPI） network analysis to obtain the key pathways and target genes for differentially expressed miRNA in regulating TNBC. TNBC patients with Qi-deficiency constitution were treated with Huadu Sanyinfang for three years after they completed the standard Western medical treatment. The peripheral blood of the patients was sampled before and after medication for detecting gene expression in the key pathways.ResultThe comparison between patients with Qi-deficiency constitution and those with moderate constitution revealed 49 differentially expressed miRNAs （16 up-regulated and 33 down-regulated）， which regulated 1 445 TNBC target genes. As demonstrated by PPI and KEGG pathway enrichment analysis， the key genes were mainly tumor protein p53 （TP53）， Akt1， epidermal growth factor receptor （EGFR）， mitogen-activated protein kinase 3 （MAPK3）， vascular endothelial growth factor A （VEGRA）， and tumor necrosis factor （TNF）. The key pathways included PI3K/Akt， MAPK， and RAS signaling pathways. A total of 11 TNBC patients with qi-deficiency constitution were enrolled. Compared with the situations before treatment， the expression levels of p105 subunit of NF-κB （NF-κB1） and Akt1 in the PI3K/Akt signaling pathway were down-regulated after medication， while the levels of catalytic subunit alpha of PI3K （PIK3CA） and B-cell lymphoma-xL （Bcl-xL） were up-regulated. The differences in NF-κB1 and Akt1 expression were statistically significant.ConclusionHuadu Sanyinfang is able to affect the gene expression of PI3K/Akt/NF-κB signaling pathway in TNBC patients with Qi-deficiency constitution. Specifically， it down-regulates NF-κB1 and Akt1 expression and up-regulates PIK3CA and Bcl-xL.
Keywords：Huadu Sanyinfang;triple-negative breast cancer （TNBC）;network pharmacology;phosphatidylinositol 3 kinase（PI3K）/protein kinase B（Akt）/transcription factor nuclear factor-κB （NF-κB）signaling pathway
Abstract：In view of the current controversy in the clinical evaluation of the terms of 'prohibition' 'contraindication'， and 'use with caution'， the present study summarized their evaluation elements to provide references for the classification of 'prohibition' 'contraindication'， and 'use with caution' and clinical rational medication of drugs. Based on the ancient and modern representative traditional Chinese medicine （TCM） literature， such as the records on herbal medicines and prescriptions， medical classics， pharmacopoeia， clinical monographs， and papers， this study proposed the evaluation elements and the underlying ideas of 'prohibition' 'contraindication'， and 'use with caution' around the risks and benefits of medication. The results indicate that the evaluation elements of 'prohibition' 'contraindication'， and 'use with caution' include TCM property，syndrome，symptom， TCM compatibility，dosage，and treatment course. When evaluating 'prohibition' 'contraindication'， and 'use with caution' of TCM under specific conditions of medication，we can determine the properties of prohibited or contraindicated drugs prior to figuring out the differences in 'prohibition' 'contraindication'， and 'use with caution'. It is feasible to evaluate the clinical 'prohibition' 'contraindication'， and 'use with caution' in TCM from Chinese medicine， body， and the clinical medication， which are correlated with each other in the practice implementation.
Keywords：prohibiting，avoiding or using caution;element;traditional Chinese medicine;ancient and modern documents
Abstract：Autism spectrum disorder （ASD） is a common neurodevelopmental disorder in childhood， whereas there is no specific medicine at present. There are more and more researches on the treatment of ASD with traditional Chinese medicine（TCM） which the curative effect is reliable. The heart and spleen are the main viscera for the treatment of ASD， but there is still a lack of in-depth analysis of the mechanism of TCM. In order to explore the relationship between the core symptoms of ASD and the heart and spleen， this article specifically explores the theoretical origins of the heart and the spleen in the formation of the core symptoms of ASD， and to clarify the role of the heart and spleen in the occurrence and development of the two core symptoms of ASD from the perspective of TCM. In view of social communication and communication obstacles， the author puts forward and explains the language problems of children with ASD based on the functions of the heart and spleen， the theory of the viscera， the ascription of the meridians， and the classics. The mechanism of the heart and spleen in TCM about the failure of the spleen， the loss of the heart， and the endogenous phlegm. Aiming at the mechanism of the stereotyped symptoms of abnormal behaviors in children with ASD， this paper proposes and explains the TCM mechanism of constant deficiency of the spleen and dereliction of duty， leading to loss of mind， heart and spleen injury， and finally a series of stereotypes and strange syndromes due to lack of spirit. Through the analysis and excavation of TCM theory， it explores theoretical basis for ASD from the theory of heart and spleen， with a view to preliminarily constructing the theoretical framework of TCM syndrome differentiation and treatment of the deficiency of both the heart and spleen， and provide theoretical reference for TCM syndrome differentiation and treatment of ASD. The treatment of ASD from the differentiation of symptoms and signs of the heart and spleen is supported by a strong theoretical basis of TCM， and the rationale， law and prescriptions are complete， which may be the direction of screening effective TCM prescriptions for the treatment of ASD in the future.
Keywords：autism spectrum disorder;traditional Chinese medicine;treatment of the heart and spleen;theoretical discussion
Abstract：Endoplasmic reticulum （ER） is an important organelle responsible for protein， steroid， lipid and carbohydrate synthesis and calcium-dependent signal transduction in eukaryotic cells. ER homeostasis is essential for normal cell function. ER homeostasis imbalance can induce ER stress （ERS）， which participates in the occurrence and development of diseases of the digestive system， respiratory system， circulatory system， nervous system， reproductive system， and endocrine system， and affects body health. Among various diseases， cancers seriously endanger people′s health due to its high mortality rate， disability rate， and recurrence rate. Due to the survival characteristics of unlimited proliferation， tumor cells are often exposed to various internal and external stimuli such as hypoxia， ischemia， excessive proliferation， and starvation， which destroy intracellular protein balance and induce ERS to some extent for survival. ERS plays a major role in various tumors and has dual functions in the survival of tumor cells： promoting the survival of tumor cells by activating a series of adaptive responses， while inducing ERS-related apoptosis pathways， so as to promote tumor cell death and inhibit tumor growth and invasion. As multiple functions of ERS in tumors are reported， many scholars have tried to intervene in the progress of tumors from the perspective of ERS. The therapeutic effect of traditional Chinese medicine （TCM） on tumors has been widely recognized. TCM can participate in the regulation of tumors from many aspects， including ERS， chemoradiotherapy resistance， gastrointestinal adverse reactions caused by chemotherapy， postoperative recurrence and metastasis. Since there are few reports on the antitumor effect of TCM from the perspective of ERS， this paper expounds the influence of ERS on tumorigenesis and development and the progress of TCM intervention in tumor through ERS， in order to provide a new direction for tumor treatment.
Keywords：endoplasmic reticulum stress;unfolded protein response;apoptosis;invasion and metastasis;traditional Chinese medicine
Abstract：Ferroptosis is a new type of cell death caused by abnormal accumulation of iron-dependent reactive oxygen species （ROS） and imbalance of redox with the participation of iron ions. In recent years， studies have found that ferroptosis is associated with various diseases and can especially regulate the development of tumors. Chinese medicine has unique advantages in tumor prevention and treatment. How to use ferroptosis theory to guide the prevention and treatment of cancer and other tumor diseases by Chinese medicine is a new research hotspot. This paper summarizes the proposal， action mechanism， and signaling pathway of ferroptosis， its application in tumor therapy， and the research on the activity of Chinese medicine based on ferroptosis. Results found that the occurrence of ferroptosis is related to iron metabolism， lipid ROS metabolism， and other signaling pathways and gene expressions. Ferroptosis can regulate tumor initiation and development， treatment， and tumor immunity， which provides strategies for tumor treatment and anti-tumor drug development. By analyzing the biological activity of Chinese medicine against ferroptosis， we found that Chinese medicines （Scutellariae Radix， Puerariae Lobatae Radix， Astragali Radix， Ginkgo， Epimedii Folium， Artemisiae Annuae Herba， and Salviae Miltiorrhizae Radix et Rhizoma）， Chinese herbal compounds （ Naotaifang， Si Junzitang， and Shenmai injection）， and Chinese medicine effective components （baicalein， dihydroartemisinin， puerarin， piperlongumine， luteolin， and quercetin） can exert antitumor and other biological activities by regulating ferroptosis. Therefore， Chinese medicine has great potential in preventing and controlling tumors and other diseases by regulating ferroptosis. This paper provides theoretical basis and research ideas for the in-depth study of ferroptosis theory and guides the prevention and treatment of tumor diseases by Chinese medicine.
Keywords：ferroptosis;signaling pathway;mechanism;Chinese medicine;tumor prevention and treatment
Abstract：Acute pancreatitis （AP） is one of the most common digestive system diseases in clinic. Its pathogenesis is complex and has not yet been fully clarified. It easily progresses to severe AP if the treatment is not provided in time， and the resulting condition is dangerous with high mortality. Intestinal mucosal barrier （IMB） injury is the key link leading to the aggravation of AP. The IMB injury in the late stage of AP promotes the translocation of harmful intestinal bacteria， the entry of bacteria and the produced endotoxins into blood circulation triggers endotoxemia and enterogenous infection，causing multiple organ failure and even death. Western medicine has limitations in the treatment of IMB injury induced by AP. By contrast， Chinese medicine has been proved effective and reliable in repairing the IMB injury induced by AP through oral administration and external application，and has been widely recognized by physicians and patients. AP falls into the categories of "precordial pain due to spleen disorder"， "thoracic accumulation"， and "pancreas-heat syndrome" in traditional Chinese medicine. The main causes of AP are excessive intake of sweet and greasy food， improper diet， and cholelithiasis， which lead to damp-heat accumulation in the middle energizer， stagnation of spleen and stomach， and obstruction of fu-organ intestine. Therefore， dredging the interior， purging， clearing heat， removing toxin， moving Qi， and activating blood should be emphasized in treatment. According to the related literature both in China and abroad in the past five years， this paper summarized the action mechanisms of Chinese medicine in the treatment of AP-induced IMB injury as follows： It protects the mechanical barrier by improving intestinal microcirculation disorders， relieving intestinal ischemia-reperfusion injury and oxidative stress response， reducing the release of inflammatory mediators and cytokines， and inhibiting the apoptosis of intestinal epithelial cells. It restores the chemical barrier by promoting gastrointestinal functional recovery and shortening enteral nutrition time. It improved the biological barrier by regulating intestinal microecological imbalance. It reinforces the immune barrier by adjusting the level of immune cells. This paper reviewed the characteristics of IMB injury in AP as well as its therapeutic principles and mechanisms with Chinese medicine， aiming to provide a theoretical and scientific basis for the in-depth study and rational application of Chinese medicine for the treatment of IMB injury in AP.
Keywords：acute pancreatitis;intestinal mucosal barrier;traditional Chinese medicine;mechanisms;review
Abstract：Polydatin， a polyphenolic compound， is the main active component of Chinese medicine Polygoni Cuspidati Rhizoma et Radix and has a variety of pharmacological activities. In recent years， there are more studies on the pharmacological effects and mechanisms of polydatin. Modern pharmacological studies show that polydatin has protective effects on the nervous system， cardio-cerebral vascular system， and respiratory system， and also has significant effects on the liver， kidney， lung， and other organs. Its effect of regulating blood glucose and blood lipid on atherosclerosis is significant， and the anti-fibrosis effect is significant on the liver and kidney. Polydatin can inhibit many types of tumor cells， suppress proliferation and induce apoptosis of tumor cells. Polydatin can also resist inflammation and radiation， protect bone marrow， and promote wound healing. Based on the literature on the pharmacological effects of polydatin， the authors found that the single pharmacological mechanism of polydatin is often regulated by multi-target proteins and multiple pathways， but the most of action targets are unclear， which needs to be further investigated. This study summarized the research progress on the pharmacological action and mechanism of polydatin in the past five years and put forward some suggestions on its present research situation and future research direction to broaden the research ideas of researchers and speed up the identification of the targets of its pharmacological effect. This study is expected to provide a scientific theoretical basis for the further development and utilization of polydatin.