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Issue 9 ， 2021
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Mechanism of Banxia Xiexintang on Rats with Chronic Gastritis Based on Quantitative Proteomics
Abstract：ObjectiveTo study the biological basis of Banxia Xiexintang against chronic gastritis by using quantitative proteomics.MethodThe experimental rats were divided into normal group，chronic gastritis model group，and Banxia Xiexintang group. The chronic gastritis model was established four weeks later by gavage with 56% ethanol. After treatment，the stomach tissues were stained with hematoxylin and eosin （HE） to observe the histopathological damage and improvement of gastric tissue in each group. The protein in gastric tissue was extracted. The differential proteins among different groups were studied by quantitative proteomics using tandem mass spectrometry tag（TMT），and the key differentially expressed proteins（DEPs） were verified by Western blot.ResultA total of 4 452 proteins were identified from rat stomach tissues，of which 318 proteins were different between the model and the normal group. After the intervention of Banxia Xiexintang，compared with the model group，there were a total of 258 differential proteins， which were mainly enriched in cell killing，nucleoid and hijacked molecular function. Kyoto encyclopedia of genes and genomes（KEGG） pathway enrichment included tricarboxylic acid（TCA） cycle，steroid hormone biosyntheis，and Retrograde endocannabinoid signaling，as well as phosphatidylinositol 3-kinase（PI3K）/protein kinase B（Akt），nuclear transcription factor-κB（NF-κB） signal pathways. Western blot verification found that 14-3-3 theta，Tenascin-C，ntercellular adhesion molecule-1（ICAM-1），stem cell factor（SCF），Caspase-3，GTPase of the Immunity-associated protein 4（GIMAP4） and mitochondrial pyruvate carrier 1（Mpc1） might be the crucial proteins for the treatment of chronic gastritis.ConclusionThe mechanism of Banxia Xiexintang in the treatment of chronic gastritis involves energy metabolism，hormone regulation，inflammation and immune processes. The target proteins found by differential proteomics and the signaling pathways may be the biological basis of Banxia Xiexintang in the treatment of chronic gastritis.Keywords：Banxia Xiexintang;chronic gastritis;proteomic;mechanism- 37
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02 -
Wuhutang Alleviates Respiratory Syncytial Virus-induced Asthma in Mice by Inhibiting STAT3 Protein and Regulating Dendritic Cell Autophagy
Abstract：ObjectiveTo investigate the therapeutic mechanism of Wuhutang on respiratory syncytial virus （RSV）-induced asthma in mice and its influence on the expression of signal transducer and activator of transcription 3 （STAT3） in lung tissue.MethodOne hundred female BALB/c mice of SPF grade were randomly divided into a normal group and an experimental group. After successful modeling via aerosol inhalation of RSV and ovalbumin （OAV）， the mice in the experimental group were further randomized into the following seven groups： model， positive control （dexamethasone， 1.82 mg·kg-1）， STAT3 inhibitor （STATTIC， 3.75 mg·kg-1）， STAT3 inducer （colivelin， 1.0 mg·kg-1）， and low-， medium-， and high-dose （1.6， 3.2， and 6.4 g·kg-1， respectively） Wuhutang groups. The corresponding drugs were administered for two weeks， followed by the detection of airway reactivity using a small animal ventilator， the pathological changes in lung tissue， mucus secretion by goblet cells and collagen deposition in airway were observed by hematoxylin-eosin （HE）， periodic acid-Schiff （PAS） and Masson staining， the serum levels of interleukin-6 （IL-6）， IL-10， and IL-17 were detected by enzyme-linked immunosorbent assay （ELISA）. The mRNA expression levels of TGF-β1 and α-SMA in lung tissue were detected by fluorescence-based real-time polymerase chain reaction （Real-time PCR）， autophagosomes present in lung tissue were examined by transmission electron microscopy， the protein expression levels of ATG5 and SQSTM1 in dendritic cells （DCs） and STAT3 and p-STAT3 in lung tissue were detected by Western blot.ResultThe airway reactivity of the model group was enhanced in contrast to that in the model group （P<0.01）， manifested as inflammatory cell infiltration around the lung tissue， excessive metaplasia of goblet cells， and extensive deposition of airway collagen， the expression levels of serum IL-6 and IL-17 were increased （P<0.01）， while that of IL-10 declined （P<0.01）， the mRNA expression levels of TGF-β1 and α-SMA were elevated （P<0.01）， the number of autophagosomes in the lung tissue increased. The protein expression levels of ATG5， STAT3， and p-STAT were up-regulated， while that of SQSTM1 was down-regulated （P<0.01）. Compared with the model group， Wuhutang and STATTIC significantly reduced the airway hyperresponsiveness of asthmatic mice （P<0.05， P<0.01）， alleviated RSV-induced pathological changes in lung tissue， reduced the contents of serum IL-6 and IL-17 （P<0.01）， increased serum IL-10 and ATG5 in DCs （P<0.01）， down-regulated the mRNA expression levels of TGF-β1 and α-SMA as well as the protein expression levels of SQSTM1， STAT3 and p-STAT3 （P<0.05，P<0.01）， and elevated the number of autophagosomes.ConclusionWuhutang relieves airway inflammation， improves airway remodeling and reduces airway hyperresponsiveness in RSV-induced asthmatic mice by inhibiting STAT3 protein and up-regulating DC autophagy in lung tissue.Keywords：Wuhutang;asthma;respiratory syncytial virus;dendritic cells （DCs）;autophagy;signal transducer and activator of transcription 3 （STAT3）- 18
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02 -
Effect and Mechanism of Didangtang on NLRP3 Inflammasomes in Diabetic Cardiomyopathy Mice
Abstract：ObjectiveTo investigate the effects of different doses of Didangtang on myocardial inflammatory lesions in diabetic mice.MethodSixty C57BL/6J mice were randomly divided into normal group （n=10） and model group （n=50）. The diabetic mice in the model group were established by intraperitoneal injection of high-fat diet combined with streptozotocin （STZ）. After model reproducing， the mice were fed with high-fat diet. After 8 weeks， the cardiac function of the mice was detected by using an ultrasound imaging platform. If the cardiac function decreased， the diabetic cardiomyopathy mice were modeled successfully. The nonmodel mice were eliminated， and finally 40 model mice were modeled. The rats in the model group were randomly divided into model group， low， medium and high dose of Didangtang group（1.5，3，6 g·kg-1） and simvastatin group（0.001 5 g·kg-1） according to heart function， with 8 rats in each group. The cardiac function of mice was detected by ultrasound imaging platform， fiber bragg grating（FBG）， triglyceride（TG） and total cholesterol（TC） were detected by automatic biochemical analyzer， hematoxylin-eosin （HE） staining was used to observe the pathological changes of myocardium， and the levels of NOD-like receptor3（NLRP3）， thiomdoxin interaction protein（TXNIP）， cysteinyl aspartate specific proteinase-1（Caspase-1） and Interleukin-1β（IL-1β） in myocardial tissue， as well as the content of reactive oxygen species（ROS） were detected by Western blot.ResultCompared with the normal control group， the levels of FBG， TC and TG in the model group significantly increased （P<0.01）； the values of EF and FS significantly decreased （P<0.01）； the expression of ROS significantly increased （P<0.05）， and the expressions of NLRP3， TXNIP， Caspase-1 and IL-1β in the myocardial tissue significantly increased （P<0.05）. Compared with the model group， the levels of FBG， TC and TG in the middle and high dose groups of Didangtang and simvastatin groups significantly decreased （P<0.05）； the EF and FS in each dose group and simvastatin group improved （P<0.05）， and the change in the middle dose group was more obvious （P<0.05）. HE staining showed that Didangtang could improve the pathological changes of myocardial tissue in mice， the ROS expression levels of mice in each dose group of Didangtang and simvastatin group significantly reduced， especially in the middle dose group， the expression levels of NLRP3， TXNIP， Caspase-1 and IL-1β in each dose group significantly decreased， and the effect of middle dose of Didangtang on reducing expressions of NLRP3， TXNIP and Caspase-1 in myocardial tissue was more obvious， the effect of high dose of Didangtang on reducing the expression of IL-1β in myocardial tissue was more obvious.ConclusionDidangtang can improve myocardial inflammatory lesions in diabetic cardiomyopathy mice by inhibiting the activation of NLRP3 inflammasome.Keywords：Didangtang;diabetic cardiomyopathy;NLRP3 inflammasome;inflammatory response;oxidative stress- 18
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02 -
Da Chaihutang Protects Liver Mitochondrial Function of Nutritionally Obese Rats via CREB/PGC-1
α PathwayAbstract：ObjectiveTo observe the effects of Da Chaihutang on Cyclic adenosine monophosphate （cAMP）-response element binding protein （CREB）/peroxisome proliferator-activated receptor-gamma coactivator-1 alpha （PGC-1α） pathway in nutritionally obese rats and the protective mechanism on liver mitochondria.MethodA total of 120 8-week-old male SD rats were randomly divided into a control group （n=20） and an experimental group （n=100）. The rats in the control group were fed on a normal diet， while those in the experimental group were administered with a high-fat feed. Successfully modeled rats were randomly divided into a model group， a positive drug （metformin） group， and low-， medium- and high-dose Da Chaihutang groups （4.25， 8.5， and 17 g∙kg-1， respectively）， with 20 rats in each group. After treatment with Da Chaihutang， the body weight， Lee's index， liver mitochondrial membrane potential and mitochondrial ultrastructure， PGC-1α expression and CREB phosphorylation of each group were measured and compared.ResultCompared with the control group， the model group showed increased body weight and Lee's index （P<0.01）， whereas decreased mitochondrial membrane potential， PGC-1α expression， and CREB phosphorylation level （P<0.01）. As compared with the model group， Da Chaihutang significantly reduced the body weight and Lee's index of obese rats （P<0.05， P<0.01）， enhanced liver mitochondrial membrane potential （P<0.05， P<0.01） to protect the integrity of mitochondrial structure， up-regulated PGC-1α expression and promoted CREB phosphorylation （P<0.05， P<0.01）.ConclusionDa Chaihutang protects the structure and function of mitochondria and inhibits weight gain in obese rats by activating the CREB/PGC-1α pathway.Keywords：Da Chaihutang;nutritionally obese rat;mitochondria;cAMP-response element binding protein （CREB）/peroxisome proliferator-activated receptor-gamma coactivator-1 alpha （PGC-1α） pathway- 28
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02 -
Analysis on Evolution of Ancient Documents of Xiehuangsan
Abstract：Xiehuangsan， derived from QIAN Yi's Key to Therapeutics of Children's Diseases， consists of 5 medicines， namely Gypsum Fibrosum，Gardeniae Fructus，Saposhnikoviae Radix，Pogostemonis Herba and Glycyrrhizae Radix et Rhizoma. It is used to treat children with spleen heat and tongue scratching. With the clinical use of later generations of physicians，the scope of diseases and syndromes of this prescription was gradually expanded，including aphthous bad breath，dry lips，yellow eyes，and sweet mouth. Modern doctors used this prescription to treat children with anorexia，constipation，allergic purpura，tic disorder， and other diseases. At present，more and more attention has been paid to the research of classical famous prescriptions. At the same time，the application of classical famous prescriptions of traditional Chinese medicine（TCM） must be researched and verified in ancient literature. Therefore，it has become important contents in the study of classic prescriptions that researching the source of prescriptions from the ancient books，combing and analyzing literature，and studying the evolution rules of indications，preparations，methods of administration and taboos.The author searched a variety of ancient Chinese medicine databases and collected the relevant documents related to Xiehuangsan in ancient medical books. A total of 242 pieces of relevant ancient document data were obtained，involving 131 types of ancient Chinese medicine books. Through combing the relevant records of historical documents，this paper analyzes and researches the historical evolution of Xiehuangsan，the source and composition of the prescriptions，the indications，the dosage，the textual research of Chinese herbal medicine and the determination of the basis，and the method of prescription preparation and administration，etc. The historical changes of Xiehuangsan and its internal relations are expected to provide literature references and theoretical basis for the modern development and research of Xiehuangsan.Keywords：Xiehuangsan;spleen and stomach Fuhuo;historical evolution;Key to Therapeutics of Children's Diseases- 9
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02 -
Textual Research on Ancient and Modern Literature of Classical Prescription Wandaitang
Abstract：The Wandaitang，recorded in an ancient medical book named Fu Qingzhu's Obstetrics and Gynecology，is one of the Classical Prescriptions. The detailed discussion and record on Wantaitang by later medical scholars can be also regarded as the inheritance and development of the original academic thought from Fu Qingzhu's Obstetrics and Gynecology. By referring to recent literatures，we have found that there are a few reports on Wandaitang from the perspective of ancient literature，but such reports are not systematic or comprehensive enough. Under the premise of inheriting but not rigidly adhering to the ancients，investigation and analysis would be made in this paper from the aspects of the origin，efficacy，dose conversion between ancient and modern uses，usage and dosage form，modern clinical application，contraindications and others based on ancient and modern literatures，in order to further promote the textual research work on Wandaitang and provide reference for its secondary research and development. The research results showed that the origin of Wandaitang can be traced back to the Bianzhenglu（Collecting Record of Differentiation of Symptoms and Signs），and its effect can be summarized as invigorating Qi and spleen，soothing liver-Qi stagnation，elevating yang and arresting leucorrhoea. In view of the great difference between the dose of the original prescription of Wandaitang and the recommended dose in the pharmacopoeia，the author recommended that the clinical dosage of Wandaitang should be 30 g Atractylodis Macrocephalae Rhizoma，30 g Dioscoreae Rhizoma，6 g Ginseng Radix et Rhizoma，15 g Paeoniae Radix Alba，10 g Plantaginis Semen，10 g Atractylodis Rhizoma，3 g Glycyrrhizae Radix et Rhizoma，3 g Citri Reticulatae Pericarpium，3 g Schizonepetae Spica Carbonisata，and 3 g Bupleuri Radix. The main indications included leucorrhea，vulva pruritus，diarrhea，stranguria，anorexia，eczema，vertigo and so on. This prescription should be used with caution in pregnant women or those with excessive phlegm and dampness，Qi stagnation of spleen and stomach，or deficiency of Yin-fluid and blood. The research and development of Wandaitang compound preparation can be inclined to the direction of compound granules，to give full play to its clinical value and market value. The above studies，based on the textual research of ancient and modern literatures，are of great significance for clarifying the origin and clinical application of Wandaitang，and provide a new idea and basis for the secondary development of Wandaitang.Keywords：classical prescriptions;Wandaitang;origin;efficacy;dosage;contraindication- 12
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02
Classic Prescriptions
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Protective Effect of Tongluo Shenggu Capsule on Function Damage of Human Umbilical Vein Endothelial Cells Induced by Glucocorticoid
Abstract：ObjectiveTo observe the effect of Tongluo Shenggu capsule （TLSGC） on glucocorticoid-induced vascular endothelial cell functional damage， and to preliminally explore the mechanism of action through MEK-ERK signaling pathway.MethodThe blood vessel of aorta rings of normal SD rats were induced in vitro intervention with methylprednisolone sodium succinate （MPS， 0.04 g·L-1） and/or vascular endothelial growth factor （VEGF， 20 μg·L-1）， and were treated with TLSGC（12.5， 25， 50 μg·L-1） continuously for 5 days to observe the number， length and area of microvascular ring buds．In addition， human umbilical vein endothelial cells （HUVEC） induced by VEGF（20 μg·L-1） were added into MPS（0.04 g·L-1） and TLSGC （12.5， 25， 50 μg·L-1） were added. Then， Transwell migration， Transwell invasion and lumen formation experiments were used to detect the migration， invasion and lumen formation ability of HUVEC， respectively. The content of nitric oxide（NO） in the cell supernatant was detected by nitrate reductase method， the content of endothelin 1（ET-1） in the cell supernatant was detected by dry powder method. Moreover， the protein contents of vascular endothelial growth factor receptor 2 （VEGFR2）， extracellular signal-regulated kinase （ERK）， phospho-extracellular signal-regulated kinase （p-ERK）， mitogen extracellular kinase1（MEK） and phosphorylated mitogen extracellular kinase1（p-MEK） in the cells were determined by Western blot.ResultCompared with the normal group， MPS could significantly inhibit the number， length and area of VEGF-induced rat thoracic aortic ring microvessels， HUVEC cell migration， invasion and lumen formation ability. It could reduce NO content and increase ET-1 content. MPS could also significantly reduce the protein content of VEGF-induced VEGFR2， p-MEK and p-ERK in HUVEC（P<0.05，P<0.01）. Compared with the model group， TLSGC could dose-dependently increase the number， length and area of MPS-induced abnormally reduced rat thoracic aortic ring microvessels， promote MPS-induced abnormally decreased HUVEC cell migration， invasion and lumen formation ability. It could increase the protein contents of NO， VEGFR2， p-MEK and p-ERK in HUVEC， and reduce abnormally increased ET-1 content（P<0.05，P<0.01）.ConclusionTLSGC has a protective effect on the damage of angiogenesis and secretion of vascular endothelial cells induced by glucocorticoid， and the mechanism may be related to the activation of MEK/ERK signaling pathway.Keywords：Tongluo Shenggu capsule;methylprednisolone sodium succinate （MPS）;human umbilical vein endothelial cells;cell function;angiogenesis- 11
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02 -
Effect of Gecko Extract on Neuroinflammation in Rats with Reserpine-induced Depression via TLR4/NF-
κ B PathwayAbstract：ObjectiveTo investigate the effects and mechanism of Gecko extract for treatment of depression in rats.MethodThe depression rats were induced by intraperitoneal injection of reserpine （0.5 mg·kg-1）. The successfully modeled rats were randomly divided into model group， fluoxetine group （1.8 mg·kg-1）， high dose and low dose groups of Gecko extract （12， 6 g·kg-1）. The rats were given corresponding dose of drugs once a day for 10 days. After administration， the levels of neurotransmitters and inflammatory factors in serum and prefrontal cortex of rats were detected by enzyme-linked immunosorbent assay （ELISA）. The cell changes in hippocampal tissues were observed by hematoxylin-eosin （HE） staining. The mRNA levels of interleukin-6 （IL-6）， nuclear factor-κB （NF-κB）， and tumor necrosis factor （TNF-α） in the hippocampus of rats were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The protein levels of Toll-like receptor 4 （TLR4） and NF-κB in hippocampal tissues of rats were detected by Western blot.ResultCompared with the normal group， Gecko extract significantly shortened the immobility time of tail suspension and swimming in mice. Compared with model group， Gecko extract significantly reduced blepharoptosis and retention time in circles for the rats （P<0.05）， increased the levels of 5-hydroxytryptamine （5-HT） and dopamine （DA） in serum （P<0.05）， decreased the levels of Monoamine oxidase （MAO）， IL-6， and TNF-α in serum （P<0.05） and prefrontal cortex （P<0.05）， decreased the mRNA levels of inflammatory cytokines IL-6， NF-κB and TNF-α and the protein expressions of TLR4 and NF-κB in the hippocampus of rats （P<0.05， P<0.01）， and improved the pathological symptoms of the hippocampus.ConclusionGecko extract can significantly alleviate the pathological damage of depression and improve the symptoms of depression， and its mechanism may be due to inhibiting TLR4/NF-κB signaling pathway and reducing the expression of NF-κB， IL-6 and other inflammatory factors in the hippocampus of rats.Keywords：Gecko;depression;Toll-like receptor 4 （TLR4）;nuclear factor-κB （NF-κB）;inflammation- 13
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02 -
Effect of Arsenic in Realgar on Rat Ability in Novel Objects Recognition Test
Abstract：ObjectiveTo determine the contents of inorganic arsenic（iAs），monomethylarsonic acid（MMA） and dimethylarsinic acid（DMA） in brain tissues and blood by using hydride generation-cold trap-atomic absorptionspectrometry（HG-CT-AAS）， and to explore the toxic effects of Realgar on central nervous system of rats.MethodThe 96 Wistar rats were randomly divided into 4 groups：normal control group，0.3，0.9 and 2.7 g·kg-1 Realgar groups. They then received intragastric administration for 14，28 and 42 days respectively， so a total of 12 groups were formed， with 8 animals in each group. The normal group was given the same dose of sodium carboxymethyl cellulose （CMC-Na） by gavage. The contents of iAs，MMA and DMA in blood and brain tissues were determined by HG-CT-AAS. The novel object recognition test was conducted to observe the learning and memory ability of rats. The changes of hippocampal neuron ultrastructure were observed by transmission electron microscopy.ResultThere was no difference in the growth，weight and hippocampal coefficient of the experimental animals. The method of HG-CT-AAS showed a good linearity，precision，accuracy and recovery in content determination of arsenic （at various forms） in rat brain and blood. MMA and DMA were detected in the brain of realgar groups at time-dose-effect relationship. iAs，MMA and DMA were detected in the blood of Realgar groups. The nuclear membrane， mitochondria and endoplasmic reticulum in hippocampus neurons of rats were gradually damaged with the increase of Rhubarb exposure dose and time. After 14 days of exposure to Realgar，compared with the normal control group，there was no significant difference in the novel object recognition index among Realgar groups. After 28 days of exposure，only 2.7 g·kg-1 Realgar group showed statistically significant difference with the control group （P<0.05）. After 42 days of exposure， the novel object recognition index of 0.9 and 2.7 g·kg-1 Realgar groups was significantly lower than that in normal control group（P<0.05）.ConclusionThe metabolites of Realgar in rats are iAs，MMA and DMA. MMA and DMA can be accumulated in the brain tissue through the blood-brain barrier，causing the decline of the ability of learning and memory and leading to damage of hippocampal neurons.Keywords：Realgar;arsenic;hydride generation-cold trap-atomic absorptionspectrometry（HG-CT-AAS）;novel object recognition test- 13
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02 -
Effect of Bushen Huatan Prescription on Serum LPS and TLR4/MyD88/NF-
κ B Signaling Pathway in Rats with Ovariectomy-induced OsteoporosisAbstract：ObjectiveTo explore the effect of Bushen Huatan prescription on serum lipopolysaccharide （LPS） and Toll-like receptor 4 （TLR4）/ myeloid cell differentiation protein 88 （MyD88）/nuclear transcription factor-κB （NF-κB） signaling pathway in rats with ovariectomy-induced osteoporosis.MethodSixty SPF 6-month-old female rats were randomly divided into sham operation group， model group， estradiol valerate group and Bushen Huatan prescription low， medium and high dose groups.One week after modeling by bilateral ovariectomy， 8 rats in each group were selected to receive intragastric administration.The estradiol valerate group was given 0.184 mg·kg-1 by gavage， and Bushen Huatan prescription low， middle and high dose groups were given 4.7， 9.4 and 18.8 g·kg-1 by gavage， sham operation group and model group were given 0.9% saline 4 mL by gavage respectively.After 12 weeks of intervention， the rats were sacrificed for detection.Serum LPS was detected by enzyme linked immunosorbent assay （ELISA）， while protein expressions of TLR4， MyD88 and phosphorylated （p）-NF-κB p65 in bone tissue were detected by Western blot， and the mRNA expressions of TLR4， MyD88， NF-κB p65， IL-1β， and IL-6 in bone tissue were detected by quantitative real-time polymerase chain reaction（PCR）.ResultCompared with sham operation group， the serum LPS level as well as protein expression of TLR4， MyD88， p-NF-κB p65 and mRNA expression of TLR4， MyD88， NF-κB p65， IL-1β， and IL-6 significantly increased in model group（P<0.05）.Compared with the model group， serum LPS level， protein expression of TLR4， MyD88， and p-NF-κB p65， mRNA levels of TLR4， MyD88， and NF-κB p65 in bone tissues as well as downstream inflammatory factors IL-1β， IL-6 mRNA expression decreased to different degrees in estradiol valerate group and Bushen Huatan prescription high dose group（P<0.05）.ConclusionBushen Huatan prescription can reduce serum LPS content， regulate mRNA and protein expression of TLR4， MyD88， NF-κB p65 and p-NF-κB p65 in TLR4/MyD88/NF-κB pathway， and down-regulate mRNA levels of IL-1β and IL-6 in bone tissues to improve bone microstructure and inhibit the development of postmenopausal osteoporosis （PMOP）.Keywords：Bushen Huatan prescription;postmenopausal osteoporosis（PMOP）;lipopolysaccharide;Toll-like receptor 4 （TLR4）/ myeloid cell differentiation protein 88 （MyD88） / nuclear transcription factor （NF-κB） signaling pathway;bone microenvironment;inflammatory cytokines;osteoimmunology- 11
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02 -
Explore Effect of Jieyu Qutan Huazhuo Prescription on Gut-liver Axis of Rats with High-fat Diet Based on 16S rDNA Sequencing
Abstract：ObjectiveTo study the effect of Jieyu Qutan Huazhuo prescription（JQHP） on the gut microbiota of rats with high-fat diet，and to explore the effect of Chinese medicine on the regulation of gut microflora and the restoration of gut-liver axis balance.MethodSeventy male SPF Wistar rats were randomly divided into a normal group of 10 and a model group of 60. Mice in the normal group were fed with normal diet and mice in the model group were fed high-fat diet. After 12 weeks，the model group was randomly divided into 6 groups with 10 animals in each group，namely the model group，Xuezhikang group，Liputuo group，and low，medium and high-dose groups of JQHP. The JQHP low-dose，medium-dose and high-dose rats were intragastrically daministered with 0.4，0.8，1.6 g·kg-1，respectively， rats in Liputuo group with Liputuo 2 mg·kg-1，rats in Xuezhikang group with Xuezhikang 0.1 g·kg-1. The rats in the normal group and the model group were intragastrically administered with the same amount of distilled water. Stool were collected after continuous gavaging for 8 weeks，16S rRNA gene sequencing was performed，blood was collected from the abdominal aorta to detect blood lipids，and the liver tissue and ileum tissue were collected for hematoxylin and eosin （HE） staining for pathomorphological observation.ResultCompared with the normal group，the total cholesterol （TC），triglyceride （TG），and low-density lipoprotein （LDL-C） in the model group were significantly increased，while the high-density lipoprotein （HDL-C） was decreased （P<0.01）. Compared with the model group，TC and TG values were decreased significantly in Xuezhikang group （P<0.01），HDL-C value was increased （P<0.05），and in the Liputuo group TC and TG were decreased significantly （P<0.01）. Compared with the model group，the middle-dose group of JQHP had a certain alleviating effect on liver steatosis and could reduce the infiltration of inflammatory cells. The JQHP could improve the proliferation of lymphoid tissues in the ileal structure，and the middle-dose group has the most significant effect. The results of Shannon curve showed that compared with the normal group，the middle-dose group of JQHP increased significantly （P<0.01）. Compared with the model group，the middle and high-dose group of JQHP increased （P<0.05，P<0.01）. Compared with the middle-dose group of JQHP，the other drug group decreased （P<0.05，P<0.01）. Principal component diversity analysis（PCA） showed that the diversity and abundance in the middle-dose JQHP group were higher than those in other drug groups. In linear discriminant analysis（LDA），compared with the normal group，Bacteroidia，Ruminococcaceae，Bacteroides S24-7，and Rumenococcus UCG-005 were down-regulated in the model group（P<0.01），while the orders of Desulfovibrionales，Erysipelotrichales and Lachnospiraceae were up-regulated in the model group （P<0.05，P<0.01）. Compared with the model group，the Bacteroidia，Ruminococcaceae，Bacteroides S24-7，and Rumencoccus UCG-005 in the middle-dose JQHP group were increased （P<0.05，P<0.01），and the orders of Erysipelotrichales were decreased （P<0.01）. Compared with the middle-dose JQHP group，Bacteroidia，Ruminococcaceae，Bacteroides S24-7，and Rumencoccus UCG-005 in other drug groups were reduced（P<0.05，P<0.01），and the order of Erysipelotrichales and Lachnospiraceae were increased （P<0.05，P<0.01）.ConclusionJQHP can regulate the abundance and diversity of the gut microbiota，improve the state of liver tissue and ileum mucosa，regulate blood lipid levels，and restore the normal intestinal ecological environment. It may be related to the regulation of inflammation-related gut microbiota in order to restore the balance of the gut-liver axis，and the middle-dose JQHP group has the best effect.Keywords：Jieyu Qutan Huazhuo prescription;gut microbiota;hyperlipidemia;gut-liver axis;16S rDNA sequencing- 10
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02 -
Study on Sub-minimal Inhibitory Concentration of Tanreqing Injection in Inhibiting Hemolytic Activity of
Staphylococcus aureus α -hemolysinAbstract：ObjectiveTo study whether Tanreqing injection （TRQ） can alleviate the body injury in the process of infection by inhibiting the production and release of α-hemolysin of Staphylococcus aureus under sub-minimal inhibitory concentration， and to provide experimental basis for better guidance of clinical medication.MethodThe effects of TRQ on the minimum inhibitory concentration （MIC） and bacterial growth of S.aureus were determined firstly by microplate method and time-growth curve. The different sub-minimal inhibitory concentrations of TRQ were co-cultured with bacteria or bacterial supernatants， and then co-incubated with defibrillated rabbit blood to detect the inhibitory and neutralizing effects of TRQ on S.aureus α-hemolysin. Cell counting kit-8 （CCK-8） cell viability assay was used to detect the protective effect of TRQ on S. aureus-mediated damage to human alveolar epithelial cells （A549）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the effect of sub-minimal inhibitory concentration of TRQ on the mRNA expression of S.aureus α-hemolysin regulatory genes hla and agrA.ResultThe MIC of TRQ to S.aureus was 1/8 of the stock solution， and the sub-minimal inhibitory concentration （1/64MIC-1/16MIC） TRQ used in this study did not affect the growth of bacteria. 1/64MIC-1/16 MIC TRQ had the effect of inhibiting and neutralizing the hemolytic activity of α-hemolysin， with a protective effect on S.aureus supernatant-mediated A549 cell damage， and its inhibitory effect on α-hemolysin was closely related to the inhibition of hla and agrA mRNA expression.ConclusionThe sub-minimal inhibitory concentration TRQ can inhibit and neutralize the hemolytic activity of α-hemolysin of S.aureus， with a protective effect on A549 cell damage mediated by S.aureus infection， and its mechanism of inhibiting α-hemolysin is closely related to the interference with agr regulatory system.Keywords：Staphylococcus aureus;α-hemolysin;Tanreqing injection;sub-minimal inhibitory concentration- 7
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02
Pharmacology
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Clinical Study of Chaihu Shugansan Combined with Abdominal Acupuncture on Depression Caused by Chronic Pain
Abstract：ObjectiveTo investigate the clinical effect of Chaihu Shugansan combined with abdominal acupuncture on depression caused by chronic pain，and to explore its mechanism.MethodA total of 97 patients with depression caused by chronic pain were randomly divided into control group （49 cases） and observation group （48 cases）. Patients in both groups received routine western medicine treatment，including necessary psychological intervention and taking paroxetine. Control groupobservation groupcontrol group Patients in control group were treated with Xiaoyaowan，and patients in observation group were treated with Chaihu Shugansan combined with abdominal acupuncture. Both groups were treated for 6 weeks. The levels of serum neurotransmitters，cytokines and Hamilton depression rating scale（HAMD） before and after treatment were compared between two groups.ResultThere was no significant difference in HAMD scores of the two groups before treatment and the HAMD scores of two groups after treatment were significantly lower than those before treatment （P<0.05），and the HAMD scores in observation group were significantly lower than those in the control group （P<0.05）. There was no significant difference in the levels of serum 5-hydroxytryptamine （5-HT），norepinephrine （NE），and brain-derived neurotrophic factor （BDNF） between two groups before treatment. After treatment，the levels of serum 5-HT，NE，and BDNF in two groups were significantly higher than those before treatment （P<0.05），and the levels in observation group were significantly higher than those in control group （P<0.05）. There was no significant difference in the levels of serum interleukin-6 （IL-6），interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α） before treatment. After treatment，the levels of serum IL-6，IL-1β and TNF-α were significantly lower than those before treatment （P<0.05），and the levels in observation group were significantly lower than those in control group （P<0.05）.ConclusionOn the basis of psychological intervention and paroxetine administration，the combination of Chaihu Shugansan and abdominal acupuncture exerts their respective advantages. It treats both symptoms and root causes of depression，relieves the degree of depression，reduces the classification of depression，and regulates the levels of neurotransmitter and cellular inflammatory factors，and inhibits inflammatory response. The clinical effect is significant.Keywords：depression;Chaihu Shugansan;abdominal acupuncture;cytokine;degree of depression- 8
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02 -
Effect of Modified Shenfuwan on Lumbar Disc Herniation Due to Kidney Deficiency and Cold Coagulation
Abstract：ObjectiveTo observe short-term and long-term effect of modified Shenfuwan on lumbar disc herniation due to kidney deficiency and cold coagulation.Method100 cases were randomly divided into the control group and observation group，50 cases each. The control group were taken meloxicam tablets，7.5 mg each time，once a day and the observation group were given modified Shenfuwan，300 mL each time，twice a day. The treatment course was 3 months. The short form McGill pain questionnaire（SF-MPQ），oswestry disability index（ODI），and lumbar intervertebral disc herniation due to kidney deficiency and cold coagulation by traditional Chinese medicine （TCM ） syndrome were observed before and after treatment. Whole blood viscosity （high，medium，low），serum inflammatory factors ［tumor necrosis factor-alpha（TNF-α），interleukin-1β（IL-1β），interleukin-6（IL-6），pain mediators［endothelin-1（ET-1），prostaglandin E2（PGE2），cyclooxygenase-2（COX-2）］ were detected. The clinical efficacyand，recurrence rate followed up for 12 and 24 months was was observed. The type and time of adverse reactions were recorded during the study.ResultAfter treatment，the total effective rate in the observation group was 98.0%（49/50） higher than that 82.0%（41/50） in the control group（P<0.05）. After 12 months of follow-up，the recurrence rate in the observation group was 23.9%（11/46） lower than that 52.5%（21/40） in the control group （χ2＝5.193，P<0.05）. After 24 months of follow-up，the recurrence rate in the observation group was 43.9%（18/41） lower than that that 80.6%（29/36） in the control group（P<0.05）. Compared with the control group after treatment，SF-MPQ，ODI，TCM symptoms，whole blood viscosity，TNF-α，IL-1β，IL-6，ET-1，PGE2 and COX-2 were significantly reduced in the observation group（P<0.05）. The incidence of adverse reactions in the observation group was 2.0%（1/50） lower than that 52.0%（26/50） in the control group（χ2＝6.264，P<0.05）.ConclusionModified Shenfuwan can significantly improve the short-term and long-term clinical efficacy of patients with lumbar disc herniation due to kidney deficiency and cold coagulation syndrome，and the incidence of adverse reactions is low.Keywords：modified Shenfuwan;lumbar disc herniation;kidney deficiency and cold coagulation syndrome- 8
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02 -
Clinical Efficacy of Midified Wendantang and Xueyaping Recipe in Middle Aged and Young People with Hypertension and Phlegm Dampness Retention
Abstract：ObjectiveTo discuss the clinical efficacy of modified Wendantang combined with Xueyaping recipe in the middle aged and young people with hypertension and syndrome of phlegm dampness accumulation， and investigate its effect on metabolism.MethodOne hundred and twenty patients were divided into control group and observation group averagely. Patients in both groups got lifestyle intervention and bisoprolol maleate tablets， 5-10 mg/time， 1 time/day. Patients in observation group additionally took modified Wendantang combined with Xueyaping recipe， 1 dose/day. Patients in control group addiiotnally got placebo granules Banxia Tianma Wan， 6 g/time， 2 times/day. The treatment was continued for 12 weeks in both groups. Blood pressure was measured at home to measure the compliance rate of blood pressure during the treatment and after the treatment. Before and after treatment， 24 h mean systolic blood pressure （24 h SBP）， 24 h mean pulse pressure （24 h PP）， 24 h mean diastolic blood pressure （24 h DBP）， blood pressure variability （BPV） ［24 h systolic blood pressure standard deviation （24 h SSD）， 24 h diastolic blood pressure standard deviation （24 h DSD）， systolic blood pressure variation coefficient （nSCV）， and diastolic blood pressure variation coefficient （nDCV） were recorded，compare night coefficients］. Scores of syndrome of phlegm dampness accumulation， body mass index （BMI） and waist hip ratio （WHR） were evaluated. Levels of uric acid （UA）， total cholesterol （TC）， triglyceride （TG）， high-density lipoprotein cholesterin （HDL-C）， low-density lipoprotein cholesterin （LDL-C）， fasting blood glucose （FBG） and fasting insulin （FINS）， insulin resistance index （HOMA-IR）， homocysteine （Hcy）， Cystatin C （CysC）， angiotensin Ⅱ （Ang Ⅱ） and nuclear factor kappa B （NF-κB） were measured. In addition， the safety was evaluated.ResultCompliance rate of blood pressure in observation group was 94.74%（54/57）， higher than 80.70% （46/57） in control group （χ2=5.211， P<0.05）. Levels of 4 h SBP， 24 h DBP， 24 h PP， 24 h SSD， 24 h DSD， nSCV， nDCV， Hcy， CysC， AngⅡ， and NF-κB in observation group were all lower than those detected from control group （P<0.01）. Score of syndrome of phlegm dampness accumulation was lower than that in control group （P<0.01）. Levels of UA， TC， TG， LDL-C and HOMA-IR were lower than those in control group （P<0.05）， while level of HDL-C was higher than that detected from control group （P<0.05）.ConclusionBased on lifestyle and western medicine intervention， Wendantang combined with Xueyaping recipe can further control the blood pressure level， reduce the symptoms of phlegm dampness retention syndrome， improve blood pressure variability， improve the compliance rate of blood pressure， improve the metabolism of patients and reduce the risk factors of ASCVD in middle aged and young people with hypertension.Keywords：hypertension;middle aged and young people;phlegm dampness retention syndrome;Wendantang;Xueyaping Recipe;metabolism;blood pressure variability- 7
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02 -
Clinical Observation on Jiechang Qingre Pills Combined with Conventional Western Medicine Therapy in Improving Dampness-heat Syndrome of Large Intestine at Active Stage of Ulcerative Colitis
Abstract：ObjectiveTo observe the clinical efficacy of Jiechang Qingre pills for dampness-heat syndrome of large intestine at active stage of ulcerative colitis （UC） and investigate its effect on inflammatory factors.MethodOne hundred and eight patients with active UC were divided into observation group and control group. Both groups were treated with Mesalazine enteric-coated tablets， 2 g/times， 2 times/day， for 2 weeks. If symptoms were poorly controlled， prednisone acetate tablets would be used instead， 0.75 mg·kg-1·d-1 in 3 times by oral administration. Patients in the observation group took Jiechang Qingre pills， 10 g/time， 3 times/day before meals. Patients in the control group took Jiechang Qingre pills simulated drug， 10 g/time， 3 times/day before meals. The course of treatment was 12 weeks in both groups and the patients were followed up for 3 months. The modified Mayo score was used to evaluate disease activity. Before and after treatment， large intestine dampness-heat syndrome score， inflammatory bowel disease questionnaire （IBDQ）， mucosal histology assessment and scores of major symptoms and intestinal mucosal lesion severity were graded. The incidence of non-reactivity， hormone failure， hormone dependence， and early recurrence were recorded 2 weeks after treatment. Tumor necrosis factor-α （TNF-α）， interleukin-6（IL-6） and IL-17 levels were measured before and after treatment.ResultThe clinical effective rate in the observation group was 94.00% （47/50）， higher than 77.55% （38/49） in the control group （χ2=5.514，P<0.05）. The clinical remission rate was 82.00%（41/50） in the observation group， higher than 61.22% （30/49） in the control group （χ2=5.266，P<0.05）. The endoscopic response rate was 96.00% （48/50） in the observation group， higher than 79.59% （39/49） in the control group （χ2=6.251，P<0.05）. The rate of mucosal healing in the observation group was 90.00% （45/50）， higher than 79.59% （35/49） in the control group （χ2=5.503，P<0.05）. The scores of diarrhea， purulent stool， abdominal pain， tenesmus， hyperemia， edema， erosion and ulcer in the observation group were lower than those in the control group （P<0.01）. The rate of non-reactivity in the observation group was 16.00% （8/50）， lower than 34.69% （17/49） in the control group（χ2=4.581，P<0.05）. The hormone failure rate in the observation group was 37.50%（3/8）， lower than 64.71%（11/17）in the control group，but the difference was not statistically significant（tested by the exact probaility method）. The hormone dependence rate in the observation group was 12.50%（1/8）， lower than 23.53% （4/17） in the control group，but the difference was not statistically significant（tested by the exact probaility method）. The early recurrence rate in the observation group was 14.00% （7/50）， lower than 32.65%（16/49） in the control group（χ2=4.827，P<0.05）. The scores of Mayo， dampness and heat syndrome and Geboes index in the observation group were lower than those in the control group （P<0.01）， and the IBDQ scores were significantly higher than those in the control group （P<0.01）. The TNF-α， IL-6 and IL-17 levels of the patients in the observation group were lower than those in the control group （P<0.01）.ConclusionBased on the routine treatment of western medicine， Jiechang Qingre pills treatment for the patients with active UC can effectively induce clinical remission， alleviate inflammatory reaction， promote intestinal mucosal healing， improve clinical symptoms， quality of life and the response of treatment. Its clinical efficacy and enteroscopy efficacy are better than western medicine treatment alone， so it is worthy of clinical use.Keywords：ulcerative colitis;active stage;large intestine dampness and heat syndrome;Jiechang Qingre pills;inflammatory factors;quality of life;recurrence- 7
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02
Clinic
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Analysis on Chemical Constituents in Rhizomes of
Bergenia scopulosa by UHPLC-Q Exactive Focus MS/MS Enhanced PublicationAbstract：ObjectiveUltra-high performance liquid chromatography coupled with quadrupole/electrostatic field orbital trap high resolution mass spectrometry （UHPLC-Q Exactive Focus MS/MS） was developed to rapidly analyze and identify the chemical components in the rhizomes of Bergenia scopulosa.MethodThe 75% methanol extract of B. scopulosa rhizomes was analyzed on a Thermo Accucore aQ RP18 column （2.1 mm×150 mm， 2.6 μm） with the mobile phase of methanol （A） and 0.1% formic acid aqueous solution （B） for gradient elution （0-40 min 5%-95%A， 40-45 min 95%A）， the flow rate was 0.3 mL·min-1 and the column temperature was at 30 ℃. The information of the chemical constituents was acquired in positive and negative ion modes by heated electrospray ion source （HESI）， and the scanning range was m/z 80-1 200.ResultA total of 66 chemical constituents were identified， including 2 free amino acids， 7 bergenin derivatives， 15 flavonoids， 15 organic acids， 25 glycosides， and 2 others.ConclusionThe chemical constituents in the rhizomes of B. scopulosa can be identified systematically， accurately and rapidly by this method. Among them， 8 compounds were unambiguously identified by comparing with reference substances （succinic acid， arbutin， gallic acid， protocatechuic acid， bengenin， catechin， chlorogenic acid and caffeic acid）， 51 compounds were found from B. scopulosa for the first time and 28 compounds were found from the genus Bergenia for the first time. This paper can provide an important basis for the further material basis clarification and quality assessment of B. scopulosa.Keywords：Bergenia scopulosa rhizomes;chemical constituents;bergenin derivatives;flavonoids;organic acids;glycosides;ultra-high performance liquid chromatography coupled with quadrupole/electrostatic field orbital trap high resolution mass spectrometry （UHPLC-Q Exactive Focus MS/MS）- 16
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02 -
Analysis and Evaluation on Leaf Quality of Different
Artemisia argyi Germplasm ResourcesAbstract：ObjectiveTo analyze the quality differences among different germplasm resources of Artemisia argyi and to screen out the specific germplasm by comprehensively evaluating 14 quality traits of 100 germplasm resources.MethodGermplasm resources of A. argyi were collected from all over the country. The output rate of moxa and the content of total volatile oil in A. argyi leaves were determined，and the contents of 12 flavonoids and phenolic acids in A. argyi leaves were detected by ultra performance liquid chromatography（UPLC）. The correlation analysis，principal component analysis and clustering analysis were used to comprehensively evaluate the quality of A. argyi.ResultThere was rich genetic diversity of A. argyi germplasm resources，and the variation coefficients of 14 quality traits ranged from 25.67% to 127.34%，among which the coefficient of variation of chlorogenic acid，cryptochlorogenic acid，isoxiafotaside，isochlorogenic acid B and isochlorogenic acid A was more than 70%，with high variation. The output rate of moxa was negatively correlated with 9 quality traits，while the content of total volatile oil was positively correlated with 10 quality traits，and most of the flavonoids and phenolic acids had synergistic effects. 12 flavonoids and phenolic acids were analyzed by principal component analysis，and 4 principal components could be extracted. The highest contents of flavonoids and phenolic acids were found in S98（Hangzhou，Zhejiang province），S84（Longhui county，Shaoyang city，Hunan province），S66（Futian river town，Macheng city，Hubei province），S35 （Balihu town，Qichun county，Huanggang city，Hubei province），and S15 （Fudao town，Tangyin county，Anyang city，Henan province）. The systematic clustering analysis showed that the 100 germplasm could be divided into four groups when the euclidean distance was 8.0，with 90，3，3，3 and 4 accessions in group Ⅰ，Ⅱ，Ⅲ and Ⅳ，respectively. The germplasm resources in group Ⅱ contained the highest content of flavonoids and phenolic acids，the group Ⅲ contained the highest content of total volatile oil and the group Ⅳ contained the highest output rate of moxa.ConclusionThe leaf quality of different A. argyi germplasms is different. This study can provide the basis for the quality evaluation and variety breeding of A. argyi germplasm resources.Keywords：Artemisia argyi;germplasm resources;ultra performance liquid chromatography;flavonoids;quality;clustering analysis- 12
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02 -
Quality Evaluation of Branches of
Juglans mandshurica by HPLC-DAD Fingerprint Combined with HPLC-Q-TOF-MS/MS Characterization of ConstituentsAbstract：ObjectiveTo establish a high performance liquid chromatography （HPLC） fingerprint of branches of Juglans mandshurica and to evaluate the quality of the samples from different producing areas and in different harvest periods.MethodChromatographic separation was performed on an Agilent Poroshell 120 SB-C18 column （2.1 mm×100 mm， 2.7 μm） for gradient elution with mobile phase of 0.2% formic acid solution （A）-0.2% formic acid acetonitrile solution （B） （0-5 min， 5%-10%B； 5-25 min， 10%-16%B； 25-40 min， 16%-22%B； 40-45 min， 22%-45%B； 45-50 min， 45%-65%B； 50-52 min， 65%-100%B； 52-55 min， 100%B） at a flow rate of 0.3 mL·min-1. The column temperature was 30 ℃ and the detection wavelength was 270 nm. The quality of branches of Juylans mandshurica was evaluated by similarity evaluation， cluster analysis， principal component analysis and partial least squares-discriminant analysis. The chemical constituents of the samples were identified by HPLC coupled with quadrupole time-of-flight mass spectrometry （HPLC-Q-TOF-MS/MS）. The mass spectrometry was conducted in negative ion mode with electrospray ionization（ESI）. Data were acquired over a range of m/z 100-1 700 for MS and m/z 50-1 700 for MS/MS.ResultA total of 19 common peaks were confirmed in 40 batches of samples， and the similarity ranged from 0.430 to 0.995， of which the similarity of samples collected in spring and winter seasons （April， May and December） was greater than 0.90， while the similarity of most samples collected in summer （July to September） was low. Multivariate statistical analysis showed that the samples were divided into two groups according to the harvest time， but there was no obvious classification rule for the samples from different producing areas. The contents of most constituents in the samples collected in spring and winter were higher than those collected in summer. The result illustrated that different harvest periods had great influence on the quality of branches of J. mandshurica. Compared with the samples collected in summer， the quality of samples collected in spring and winter was better. A total of 22 peaks were proved to be the main constituents that contributed to the difference between samples collected in different seasons. A total of 83 chemical components were identified by HPLC-Q-TOF-MS/MS， including 49 tannins， 7 organic acids， 14 naphthalene derivatives， 1 flavonoid， 6 anthracene derivatives， 2 lignans， 3 diarylheptanoids and 1 saccharide. Totally 13 common peaks were identified. Of the peaks that contributed to discriminate samples collected in different season， 19 peaks were identified and most of them were tannins.ConclusionThe established HPLC fingerprint can provide useful information for the quality evaluation of branches of J. mandshurica. Tannin is the main constituents in the samples. Harvest period has great influence on the quality of branches of J. mandshurica.Keywords：branches of Juglans mandshurica;quality evaluation;fingerprint;high performance liquid chromatography （HPLC）;liquid chromatography coupled with mass spectrometry （LC-MS）;material basis;harvest periods- 9
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02 -
Effect of Removing Core on Quality of Phyllanthi Fructus
Abstract：ObjectiveThe differences of chemical compositions and pharmacological activities between the core and pulp of Phyllanthi Fructus were investigated by chemical analysis and in vitro test to explore the effect of the core on the quality of this medicinal material.MethodLiterature， medicinal material standards and market research on the appearance of Phyllanthi Fructus were conducted based on existing databases. Ultra-high performance liquid chromatography-quadrupole-electrostatic field orbital trap high resolution mass spectrometry （UPLC-Q-Orbitrap HRMS） was used to identify the constituents of the core and pulp. The analysis was performed on Thermo Scientific Accucore C18 column （2.1 mm×100 mm， 2.6 μm） with the mobile phase of 0.1% formic acid aqueous solution （A）-methanol （B） for gradient elution （0-25 min， 5%B； 25-30 min， 5%-95%B； 30-35 min， 95%-5%B）， the flow rate was 0.2 mL·min-1， heating electrospray ionization （HESI） was adopted with positive and negative ion modes， and the scanning range was m/z 100-1 500. High performance liquid chromatography （HPLC） was used to determine the contents of gallic acid， corilagin， chebulagic acid and ellagic acid in the core and pulp of Phyllanthi Fructus. Analysis was performed on Welchrom C18 column （4.6 mm×250 mm， 5 μm） with mobile phase of methanol （A）-0.05% phosphoric acid aqueous solution （B） for gradient elution （0-6 min， 5%A； 6-15 min， 5%-7%A； 15-20 min， 7%-15%A； 20-25 min， 15%-21%A； 25-31 min， 21%-22%A； 31-41 min， 22%A； 41-47 min， 22%-28%A； 47-51 min， 28%-32%A； 51-57 min， 32%-38%A； 57-70 min， 38%-45%A； 70-80 min， 45%-65%A； 80-85 min， 65%-5%A）， the detection wavelength was set at 270 nm. The antibacterial effects of the core and pulp of Phyllanthi Fructus on Escherichia coli and Staphylococcus aureus were investigated by filter paper method， and their antioxidant activities were compared by 1，1-diphenyl-2-picrylhydrazyl （DPPH） free radical scavenging assay.ResultA total of 47 compounds were identified in the core and pulp of Phyllanthi Fructus， mainly including tannins， flavonoids， phenolic acids， fatty acids， amino acids， organic acids， saccharides and glycosides， most of which were concentrated in the pulp， and the fatty acids in the core accounted for a higher proportion. The contents of gallic acid， corilagin， chebulagic acid， ellagic acid and other phenolic compounds in the pulp of 20 batches of Phyllanthi Fructus were much higher than those in the core. The results of antibacterial test showed that the core of Phyllanthi Fructus with different concentrations had no antimicrobial effect. The DPPH radical scavenging test showed that the antioxidant activity of the core ［half-inhibitory concentration （IC50）=199.632 mg·L-1］ was much less than that of the pulp （IC50=12.688 mg·L-1）.ConclusionFrom the perspectives of polyphenol content， antibacterial and antioxidant activities， it is scientific to use Phyllanthi Fructus pulp in ancient and modern times， which may be to remove the secondary parts of Phyllanthi Fructus， so as to enhance the actual utilization rate and therapeutic effect of medicinal materials. In view of the large proportion of the core of Phyllanthi Fructus and its high content of fatty acids and other components， whether or not to use it remains to be further studied in clinical application.Keywords：Phyllanthi Fructus;removing core;high performance liquid chromatography （HPLC）;antibacterial activity;antioxidant activity;Chinese medicinal materials- 6
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02 -
Analysis of Chemical Constituents in Jiechangyan Qixiao Granules Based on UPLC-Q-TOF/MSE
Abstract：ObjectiveIn order to systematically clarify the chemical composition of Jiechangyan Qixiao granules， the main chemical components in this preparation were rapidly identified and assigned by ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF/MSE）.MethodACQUITY UPLC BEH C18 column （2.1 mm×100 mm， 1.8 μm） was employed for UPLC analysis with the mobile phase of 0.1% formic acid aqueous solution （A）-acetonitrile （B） for gradient elution （0-2 min， 5%B； 2-16 min， 5%-21%B； 16-30 min， 21%-95%B； 30-33 min， 95%B； 33-34 min， 95%-5%B； 34-37 min， 5%B）. The flow rate was 0.3 mL·min-1， the column temperature was 30 ℃， and the volume of sample injection was 2 μL. Electrospray ionization （ESI） was applied for scanning under positive and negative ion modes with the scanning range of m/z 60-1 200. MSE mode was used to collect mass spectral data. The ion peaks were identified by comparing with the information of control substances， literature references and self-built database.ResultA total of 102 chemical components were separated and identified in Jiechangyan Qixiao granules， including organic acids， flavonoids and its glycosides， triterpenes， phenylethanoid glycosides， tannins， iridoid glycosides and other components， among which flavonoids and its glycosides were from Drynariae Rhizoma and Crataegi Fructus， phenylethanoid glycosides and iridoid glycosides were from Plantaginis Semen， triterpenoids and tannins were from Crataegi Fructus and Chebulae Fructus. Among the identified chemical constituents， there were 28 from Drynariae Rhizoma， 31 from Plantaginis Semen， 53 from Chebulae Fructus and 58 ingredients from Crataegi Fructus.ConclusionThe established UPLC-Q-TOF/MSE can comprehensively and rapidly analyze the chemical constituents in Jiechangyan Qixiao granules， and preliminarily elucidates the chemical composition profile of this granules， which can lay a foundation for further research on the pharmacodynamic material basis and quality control of Jiechangyan Qixiao granules.Keywords：Jiechangyan Qixiao granules;ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF/MSE）;material basis;flavonoids;iridoid glycosides;triterpenes;Chinese herbal compound- 9
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02 -
Identification of
Levisticum officinale Adulterated inAngelica sinensis by PCR-RFLPAbstract：ObjectiveTo establish a rapid method to identify Levisticum officinale adulterated in Angelica sinensis by polymerase chain reaction -restriction fragment length polymophism（PCR-RFLP）.MethodBy comparing sequences restriction sites in ribosomal DNA Internal Transcribed Spacer（ITS） of A. sinensis and L. officinale，the specific restriction site Fnu4HI of L. officinale was selected，and the primers for PCR-RFLP reaction were designed. Different A. sinensis and L. officinale were amplified by PCR. The conditions affecting the PCR-RFLP reaction，such as annealing temperature，primer concentration，cycle number and enzyme digestion reaction time，were optimized，and the accuracy of the method was investigated. The established PCR-RFLP identification method was used to investigate the applicability of L. officinale adulterated in A. sinensis with different aduleration ratios and different origins.ResultA PCR-RFLP method for identifying A. sinensis mixed with L. officinale was established. When the annealing temperature was 62 ℃ and the number of cycles was 30，when the L. officinale adulterated in A. sinensis could be digested by Fnu4H I restriction endonuclease after amplification with specific primers，and the two single DNA bands were detected between 100-500 bp，the A. sinensis were all negative. The minimum detection limit of this method for adulterated L. officinale in A. sinensis was 3%，which could be used for the detection of adulterated L. officinale in A. sinensis.ConclusionThe established PCR-RFLP identification method is sensitive and accurate in detecting whether there is L. officinale in A. sinensis，and it provides inspection reference and basis for the quality control of A. sinensis，with great significance to ensure the safety of its clinical medication.Keywords：Angelica sinensis;Levisticum officinale;polymerase chain reaction -restriction fragment length polymophism （PCR-RFLP）;admixture identification- 9
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02
Pharmacy Fundamentals
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Analysis of Medication Rules of Coronary Heart Disease with Heart Failure of Qi Deficiency and Blood Stasis Syndrome Based on Literature Research
Abstract：ObjectiveTo systematically sort out and summarize the medication rules of clinical prescriptions for coronary heart disease with heart failure of Qi deficiency and blood stasis syndrome，and to provide reference for selecting prescriptions and medications for the treatment of coronary heart disease （CHD） with traditional Chinese medicine （TCM）.MethodAll relevant literature concerning the treatment of CHD with compound TCM prescriptions for Qi deficiency and blood statis syndrome from 2000 to 2020 were retrieved from the China National Knowledge Network （CNKI），WanFang database （WanFang），and VIP journal database （VIP），and the names of prescriptions and drug components were extracted，followed by the frequency of drug use and drug category. Association rules of high-frequency drugs were analyzed by SPSS 18.0，and systematic clustering analysis was conducted by SPSS 21.0.ResultFinally，41 qualified literature articles covering 35 prescriptons and 66 drugs were included in the study. The total frequency of the drugs was 433 times. Among them，there were a total of 25 traditional Chinese medicines with a frequency of ≥5 times. The top 3 frequently used Chinese medicines were Astragali Radix （8.8%），Salviae Miltiorrhizae Radix et Rhizoma （7.2%），Chuanxiong Rhizoma （5.8%）. A total of 15 types of drugs were involved，among which tonic drugs （31.4%），blood promoting and blood stasis drugs （28.2%），and hydration and dampening drugs （7.6%） were used most frequently. The association rule analysis of traditional Chinese medicines with frequency of ≥ 5 showed that there were 13 pairs of binomial associations in 25 traditional Chinese medicines，with Astragali Radix in combination with Salviae Miltiorrhizae Radix et Rhizoma，Chuanxiong Rhizoma，Carthami Flos，et al. There were 8 groups of three associations，with Astragali Radix，Salviae Miltiorrhizae Radix et Rhizoma，and Chuanxiong Rhizoma as the main combinations. A systematic clustering analysis showed that the clustering effect was best when the 25 traditional Chinese medicines were clustered into 5 categories.ConclusionThe treatment of coronary heart disease with heart failure of Qi deficiency and blood stasis syndrome is based on replenishing qi and activating blood circulation，supplemented by warming yang，diuresis，and phlegm-resolving drugs，which can enhance the clinical efficacy. Two basic prescriptions of Buyang Huanwutang and Si junzitang are extracted. Other combinations of prescriptions and drugs can provide references for the clinical treatment of coronary heart disease with heart failure.Keywords：coronary heart disease;heart failure;Qi deficiency and blood stasis;medication rules;data mining- 13
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02 -
Generic Association Analysis on "Prescription-Formula-Dosage-Property" of Tibetan Medicine Based on Partial Order Structure and Property Theory
Abstract：ObjectiveIn view of the complexity of the "Prescription-Formula-Dosage-Property" relationship of Tibetan medicine and the outstanding common relationship，it is difficult to reveal the hidden and specific rules of clinical medication of Tibetan medicine.MethodBased on the attribute partial order structure and the vector structure model of "Ro-Nus-Zhu-Rjes"（Taste，Post-Digestive Tastes and Potency），clustering analysis and other methods and software，this study analyzed the "Prescription-Formula-Dosage-Property" relationship of 184 commonly used prescriptions in the 1995 edition of the standards issued by the Ministry of Tibetan medicine（SIMTM）.ResultAmong them，the analysis of the relationship between prescription and formula found that 11 prescriptions with the largest common attribute，such as Chebulae Fructus and Carthami Flos，were the key components of classification and compatibility，which could effectively classify the 8 kinds of prescriptions for the treatment of lung disease，tripa disease and blood fever. Among them，the san Yin and auxiliary viscera function prescriptions，such as Sanguotang powder and Liuwei Liangyao powder，had the strongest commonality. According to the analysis of relationship between formula and dosage ，the dosage of Chebulae Fructus，Carthami Flos，pomegranate seed，Phyllanthi Fructus was the highest，which suggested that these drugs were often used as primary drugs，while the Liuwei Liangyao powder，such as Amomi Fructus Rotundus and Tsaoko Fructus，had a higher frequency but a lower dose，which mainly played a role in regulating the overall drug property of the prescription and protecting the viscera. The Tibetan medicine-specific drugs including Moschus，Bovis Calculus，and Zhaxun，which were used in a high frequency but very low dose，had the effect of enhancing the drug property and guiding the affected part. According to the analysis of the relationship between dosage and property，there were many prescriptions belonging to cool nature，accounting for 75.6%. It was found that 67 prescriptions did not conform to the efficacy due to their different dosage.ConclusionThere are many common components and common usages in Tibetan medicine prescriptions. If these common associations are not treated，it will lead to the result that all diseases take these common associations as the core，but the hidden key factors cannot be solved. Therefore，it is necessary to pay attention to sensitivity and specificity at the multi-dimensional level of "prescription-formula-dosage-property"，so as to reveal the clinical medication thought of Tibetan medicine more effectively.Keywords：Tibetan medicine;Ro-nus-zhu-rjes;partially ordered structure;prescription-formula-dosage-property;relevance research- 10
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02 -
Pharmacological Mechanism of Wumeiwan in Treating Lung Metastasis of Breast Cancer
Abstract：ObjectiveTo explore the potential target and mechanism of Wumeiwan in the treatment of lung metastasis of breast cancer by network pharmacological analysis and experimental verification.MethodThe databases of active ingredients and targets of Wumeiwan were established through Traditional Chinese Medicine Systems Pharmacology（TCMSP） Database and Analysis Platform，and the targets of lung metastasis of breast cancer were established through the GeneCards database and Online Mendelian Inheritance in Man（OMIM） database，and the data of Chinese medicine targets and disease targets were matched. Cytoscape 3.6.0 software was used to establish the network analysis of traditional Chinese medicine-active ingredients-therapeutic targets，and the interaction relationship between key target proteins was analyzed by STRING database. Target gene ontology（GO） analysis and Kyoto Encyclopedia of Genes and Genomes（KEGG） signal pathway enrichment analysis were performed by using the Biological Information Annotation Database.ResultA total of 108 possible important targets for Wumeiwan in the treatment of lung metastasis of breast cancer were found，including interleukin 6（IL6），cysteine aspartate-specific protease-3（CASP3），vascular endothelial growth factor A（VEGFA），epidermal growth factor receptor（EGFR），mitogen-activated protein kinase（MAPK8）， and others. GO enrichment analysis yielded 29 cell components（CC），1 218 biological processes（BP） and 125 molecular functions（MF） related to lung metastasis of breast cancer，and KEGG enrichment analysis yielded 118 pathways related to lung metastasis of breast cancer（P<0.05），including MAPK signaling pathway and apoptosis pathway. In vitro experiments showed that cinnamaldehyde， the active ingredient of Wumeiwan， could induce apoptosis，inhibit proliferation and migration of MCF7 cells，partially validating the predicted results of network pharmacology to a certain extent.ConclusionThe therapeutic effect of Wumeiwan on lung metastasis of breast cancer may be multi-target，multi-pathway and multi-mechanism. The results of this study provide more evidence for the clinical application of Wumeiwan.Keywords：Wumeiwan;lung metastasis of breast cancer;network pharmacology;mechanism;cinnamaldehyde- 17
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02 -
Huayu Jiedu Prescription Alleviates Gut Microbiota and Fecal Metabolites in Mice with Endometriosis
Abstract：ObjectiveTo investigate the effect of Huayu Jiedu prescription （HYJDP） on gut microbiota and fecal metabolites in mice with endometriosis.MethodNormal female C57BL/6J mice were divided into normal control group （CO）， endometriosis group （EM） and Chinese medicine Huayu Jiedu decocotion group （CM）. CO and EM groups received normal saline and CM group received HYJDP by intragastric administration. Untargeted metabolomics method was used to detect metabolites in fecal supernatant of mice， and receiver operating characteristic （ROC） analysis was used to screen the differential metabolites， 16S rRNA high-throughput sequencing was used to detect the gut microbiota， and Spearman correlation coefficient was used to represent the degree of correlation between differential metabolites and intestinal flora. Lipopolysaccharides （LPSs） in intestinal wall tissue， serum and peritoneal lavage fluid were detected by enzyme-linked immunosorbent assay （ELISA）. The expression of Vimentin and E-cadherin in ectopic lesions was detected by immunohistochemistry.ResultHYJDP alleviated the disorders of fecal metabolites and gut microbiota in EMS mice， especially with the recovered levels of homoveratric acid， melilotoside C and physapubescin in fecal supernatant. In the comparison of these three factors between EM group and CO group as well as between EM group and CM group， the variable important in projection （VIP） value was both above 2， and AUC in ROC analysis was both >0.9. As compared with EM group， HYJDP restored the abundance of species such as Lachnospiraceae_NK4A136_group， Lactobacillus and Blautia （P<0.05）. In addition， the level of LPS in peritoneal fluid supernatant of EM group was significantly higher than that of CO group （P<0.05） and CM group （P<0.05）. The protein expression of vimentin and E-cadherin in endometriosis decreased significantly （P<0.05）.ConclusionHYJDP which can improve the intestinal environment and reduce the level of LPS in mice with endometriosis， is an effective drug for the treatment of endometriosis.Keywords：endometriosis;Huayu Jiedu prescription （HYJDP）;intestinal flora;metabolomics;lipopolysaccharide- 18
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02
Data Mining
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Research Progress of Transmembrane Transport of Oral Nanoparticles in Gastrointestinal Tract
Abstract：Oral nanoparticles （NPs） has gradually become a approach to improve oral bioavailability of biopharmaceutics classification system （BCS） Ⅱ， Ⅲ， Ⅳ drugs， and the transmembrane transport mechanism in the gastrointestinal tract largely depends on physicochemical characteristics of NPs. It would be beneficial to design the NPs with high transport efficiency and effectively improve the oral bioavailability of drugs by adopting a reasonable research model to analyze the transmembrane mechanism of the oral NPs and exactly reveal the relationship between the physicochemical properties and the transport mechanism of NPs. This review focused on summarizing the transmembrane approaches of oral NPs， comparing the advantages and disadvantages of the common cell models， concluding the potential interaction between the physicochemical properties and transmembrane process of NPs， and proposing the research strategy of transport mechanism based on in situ intestinal perfusion， with the purpose of discovering a suitable research model for studying the transport mechanism of different NPs， providing a basis for regulating the transport performance of the NPs to improve the oral bioavailability， and expanding the application of oral NPs in the development of new drugs.Keywords：oral nanoparticles;transmembrane transport;cell models;physicochemical properties;particle design;bioavailability- 10
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02 -
Historical Evolution and Modern Clinical Application of Qingxin Lianzi Yin
Abstract：Qingxin Lianzi Yin （QXLZY）， as an ancient classical formula for clearing the heart and nourishing the Yin， was composed of nine herbs （Scutellariae Radix， Ophiopogonis Radix， Lycii Cortex， Plantaginis Semen， Glycyrrhizae Radix et Rhizoma Praeparata Cum Melle， Nelumbinis Semen， Poria， Astragali Radix and Ginseng Radix et Rhizoma）， coming from Prescriptions of the Bureau of Taiping People's Welfare Pharmacy. It could clear away the heart-fire， promote the interaction of the heart and kidney， replenish Qi and Yin， and stop strangury-turbidity. It was used to treat gonorrhea of urination， seminal emission， restlessness， wasting-thirst and so on. At present， the usage and dosage of QXLZY and its addition and subtraction are different in clinical practice. Most of the studies just focus on its clinical efficacy， and there is few review literature reflecting its historical evolution. Based on this， this paper systematically clarified the historical evolution， composition， preparation， interpretation， function， and modern clinical application of QXLZY. This work has been explained the historical evolution of QXLZY， and found that it was wildly used in modern clinical， especially suitable for the treatment of chronic urinary system diseases. At the same time， QXLZY also had significant therapeutic effects on neurasthenia， stomatitis， diabetic nephropathy and other aspects. Through the comprehensive analysis of ancient and modern literature， this work explores the true connotation of QXLZY from the perspective of traditional Chinese medicine theory， which can point out the direction of the clinical application and positioning of this famous classical formula after it comes into the market， and also can provide reference basis for its subsequent in-depth research and development.Keywords：Qingxin Lianzi Yin;famous classical formulas;historical evolution;clinical application;Prescriptions of the Bureau of Taiping People's Welfare Pharmacy;weights and measures;Song dynasty- 16
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02 -
Research Progress on Anti-tumor Effect and Mechanism of Cucurbitacins
Abstract：Cancer is a major global public health problem. Statistics from the national cancer center of China also show that cancer has become a major disease threatening human health with increasing morbidity and mortality. The occurrence and development mechanism of cancer is complex， involving multiple stages， multiple genes and multiple signaling pathways. Conventional chemoradiotherapy and emerging targeted therapy methods are the main methods in treatment of tumor. However， the quality of life of patients as well as the sustained and effective therapeutic effect are seriously affected due to the toxic side effects and drug resistance. Therefore， it is the global focus to find safe and effective anti-cancer drugs. The research and development and application of anti-cancer herbal medicines such as paclitaxel， vinblastine， podophyllin， ginsenoside and ginseng polysaccharide have brought new hope for the treatment of cancer. Cucurbitacine from Chinese medicine cucurbitaceae plantsare is a kind of highly oxidized tetracyclic triterpene compound with extensive pharmacological effects and complex mechanism. In the family of cucurbitacines， cucurbitin B， D， E and I have been studied most frequently on anticancer effect， and in a large number of studies， they have been found to play an important role in tumor diseases of the digestive system， respiratory system， reproductive system， blood system， urinary system and so on. With significant effect in inhibiting tumor cells proliferation， blocking the cell cycle， inducing apoptosis and autophagy death， inhibiting cell migration and invasion， inhibiting tumor angiogenesis， regulating the levels of reactive oxygen species and regulating immune system， such cucurbitacins are expected to be developed as a new kind of anti-cancer drugs. The authors of this study aim to provide reference for the further research and development of new anti-cancer drugs about cucurbitines by summarizing the anti-tumor effect and mechanism of the cucurbitacins.Keywords：cucurbitacins;tumor;cell cycle;cell apoptosis;cell invasion and migration;tumor angiogenesis- 15
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02 -
Research Progress of Traditional Chinese Medicine in Improving Pathogenesis of Depression
Abstract：Depression is a kind of psychoaffective disorder，which is mainly characterized by persistent depression and mood disorders. It is related to many factors such as abnormal expression of neurotransmitters，secretion of inflammatory cytokines，changes in neuronal pathways，persistent disorder of hypothalamus-pituitary-adrenal （HPA） axis，changes of intestinal flora environment，abnormal mitochondria structure，and many other factors，which cause serious damage to human health. At present，western medicine is mainly treated with antidepressant drugs in clinical practice，which can exert short-term efficacy，but long-term use will cause adverse side effects. Traditional Chinese medicine（TCM） emphasizes the holistic view of psychosomatic integration and has rich clinical practice experience，which gives it a certain therapeutic advantages in the prevention and treatment of depression. Moreover，modern research has shown that TCM has played an important role in the prevention and treatment of depression through its multi-target，multi-mechanism and multi-pathway mode of action. In recent years，the study on TCM in treating depression has been deepening，and more and more TCM and TCM compounds have been proved to be able to regulate and improve the pathogenesis of depression and play an antidepressant role. Therefore，this paper systematically sorted out the research literature on TCM in improving the pathogenesis of depression in the past 5 years，and summarized and elaborated these literature according to the following topics：TCM affects the expression of neurotransmitters and their receptors，improves the plasticity of nerve cells and related signaling pathways，regulates the secretion of inflammatory factors and improves the disorder of HPA axis， TCM regulats intestinal flora environment to relieve depression， TCM adjusts the mitochondrial function of TCM to relieve depression. The study found that most of the TCM exert the antidepressant effect mainly by affecting the expression of neurotransmitters and improving neural signaling pathways，while the research on Chinese medicinals regulating and improving the intestinal flora environment and mitochondrial function is relatively rare，which needs further research to provide reference for better exploration of depression in the future.Keywords：traditional Chinese medicine;depression;pathogenesis;research progress- 22
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<HTML><CITATION> <Bulk Citation>Published：2021-04-02
Review
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