Abstract：Coronavirus disease-2019 (COVID-19) is highly contagious. In the early stages of the disease, the symptoms of coldness, dampness and depression in the lungs often appear, including fever, fatigue, soreness, dry cough, poor appetite, and white greasy tongue coating. During the development of the disease, the damp toxin epidemic often enters the inner and generates heat, and the damp heat epidemic toxin blocks pleurodiaphragmatic sites, the triple energizer and lungs, with manifestations of chest tightness, shortness of breath, exacerbation after exercise, or high fever without bringing down, poor appetite, bitterness in mouth, fatigue, diarrhea, loose stools, and yellow and thick tongue fur. As the pathogen can go outward or enter more deeply inside to cause death at the moment, it is the crucial time to treat the disease. In this study, Haoqin Qingdantang was used to clear dampness and heat, reconcile Shaoyang channel and recover the triple energizer, detoxify the dampness fever epidemic toxin, and block the toxin inside, with a good efficacy. This prescription focuses on smoothing the Shaoyang gallbladder channel and the triple energizer, and regards the spleen and stomach as the acquired essence. In the prescription, Erchentang reconciles the spleen and stomach, elevates clear Qi and lower turbid Qi. Bupleuri Radix is added to increase its detoxification function, and Paeoniae Rubra Radix is added to circulate the blood and prevent pathogen from the blood. When the condition improves, Sweet Wormwood Herb and talc are often withdrawn, and Codonopsis is added to nourish the spleen and stomach, to strengthening vital qi to eliminate pathogenic factor. Based on the cases of COVID-19 treated in Wuhan Jinyintan Hospital, this paper summarized some experience of applying Haoqin Qingdantang.
Abstract：ObjectiveIn the early stage, in vivo experiments of coronavirus infection and pulmonary syndrome syndrome combined with mouse model verified that Reyanning mixture has a significant therapeutic effect on human coronavirus disease-2019 (COVID-19). This study further evaluated the clinical efficacy of Reyanning mixture in the treatment of COVID-19, providing a basis for clinical medication.MethodPatients were collected from January 21, 2020 to February 24, 2020 in Xi'an No. 8 Hospital, Second Affiliated Hospital of Xi'an Jiaotong University, Yan'an Second People's Hospital and Songzi People's Hospital. 54 common-type patients who met the diagnostic criteria of COVID-19 were enrolled in this study,5 patients of them were not included in the statistical analysis because they did not meet the requirements, including 26 cases in the treatment group using Reyanning mixture on the basis of chemical drug treatment, and 23 cases in the control group receiving chemical drug treatment only. The symptom disappearance rates (throat dryness, throat pain, cough, fever, fatigue, chest tightness, runny nose, nasal congestion, headache), time to complete fever clearance (d), the nucleic acid conversion rate and time to recovery on chest CT were compared between two groups.ResultAfter treatment, except cough and fatigue, other symptoms disappeared in the treatment group, and the disappearance rate of symptoms such as dry throat, cough, fatigue, chest tightness and headache was statistically significant compared with the control group (P<0.05). The median time to complete fever clearance in the treatment group was 3 days, which was 2 days shorter than that in control group. The remission rate of chest CT was 88.46% (23/26) in the treatment group, which was higher than 73.91% (17/23) in control group. The negative conversion rate of viral nucleic acid detection was 96.15% (25/26) in treatment group, higher than 60.87% (14/23) in control group (P<0.01).ConclusionReyanning mixture can improve the clinical symptoms of COVID-19 patients, promote the improvement of chest CT. Can shorten the duration of fever.Can improve the novel coronavirus nucleic acid conversion rate, providing a basis for clinical treatment.
Abstract：Since the outbreak of coronavirus disease 2019 (COVID-19), the health authorities in various regions have formulated a group of traditional Chinese medicine (TCM) diagnosis and treatment programs based on the consensus analysis of etiology and pathogenesis of TCM experts, and recommended the guiding prescriptions for different syndromes in three stages of prevention, treatment and recovery. In order to effectively summarize the understanding of the pathogenesis of each scheme and the rules of the formulation of various prescriptions, based on 49 TCM prevention and treatment plans issued by the health administration departments, 308 TCM prescriptions were sorted by stages (151 prescriptions in the treatment period, 118 prescriptions in the prevention period and 39 prescriptions in the recovery period). The 2019-nCoV pneumonia prescriptions was further processed by using TCM inheritance platform system. The common syndromes and the frequency of commonly used Chinese medicines were summed up. The commonly used drug pairs and combinations in different stages were extracted, and their association rules were analyzed. The rules of the Chinese herbal prescription for COVID-19 were preliminarily summarized. During the treatment period, the main syndrome types are inner blocking causing collapse, epidemic toxin in the lung, cold dampness in the lung and toxic heat in the lung, involving 187 kinds of herbs, 29 kinds of herbs with single taste frequency more than 20 times.Armeniacae Semen Amarum is the most frequently used, Gypsum Fibrosum-Armeniacae Semen Amarum is the most frequently used drug-pair, the core formula is Maxing Shigan Tang, and the main pathogenesis is cold, wet and heat. There is no unified syndrome type in the prevention period, involving 119 herbs in total. There are 13 kinds of herbs with single taste frequency more than 20 times. The most frequently used is Astragali Radix. The most frequently used medicine pair is Astragali Radix-Saposhnikoviae Radix. The core formula is Yupingfeng San, and the main pathogenesis is Qi deficiency. During the recovery period, the main syndromes are deficiency of lung and spleen Qi deficiency and Qi and Yin deficiency, involving 113 herbs, 12 kinds of herbs with single taste frequency more than 10 times, Poria with the highest frequency of use, Citri Reticulatae Pericarpium-Poria with the highest frequency of drug pairing, Erchentang and Shengmaiyin with the main pathogenesis of deficiency of spleen Qi and Yin. The purpose of this study is to provide multiple references for strengthening clinical precise treatment, improving the effect of TCM treatment and promoting the management of TCM prevention and control resources.
Keywords：coronavirus disease 2019（COVID-19）;composition principles;traditional Chinese medicine inheritance platform system
Abstract：ObjectiveTo comprehensively analyze the diagnosis and treatment programs and prevention and treatment programs on tratidional Chinese medicine (TCM) for coronavirus disease-2019 (COVID-19), and to provide suggestions for further development of TCM clinical guidelines.MethodDiagnosis and treatment programs on TCM for COVID-19 pneumonia, as well as prevention and treatment programs, clinical practice guidelines and expert consensus, were retrieved till Feb 19, 2020. The information about TCM syndrome differentiation, state of disease, and TCM treatments (decoction and Chinese patent medicines) were extracted and analyzed.ResultA total of 24 TCM programs/guidelines were included containing 23 diagnosis and treatment programs or prevention and treatment programs and one rapid advice guideline. Of the 23 TCM programs, 14 programs described the classification of TCM syndromes and the stages of disease，22 programs described the composition of the TCM decoction; seven programs described how to add and subtract the herbs according to different TCM syndromes,17 programs described the weight/volume of the herbs of TCM decoctions, three programs described the decoction method，six programs described the usage and dosage of TCM decoction， two programs clarified the course of treatment; none of the 23 programs indicated the source of evidence. The TCM treatment within the rapid advice guideline was in reference to the fourth edition of the COVID-19 pneumonia TCM diagnosis and treatment program issued by the National Health Commission. A total of 41 Chinese patent medicines were recommended in 23 programs, of which 12 Chinese patent medicines were off-label recommended.ConclusionIn most TCM programs, TCM decoction and Chinese patent medicines are recommended based on TCM syndrome differentiation in combination with the state of disease, and the dosage of the TCM decoctions are clearly described. Some Chinese patent medicines in the TCM programs are off-label recommended. Expert experience and opinions are valued when developing TCM programs/ guidelines. All of these provide reference for developing TCM programs/guidelines in future.
Keywords：coronavirus disease 2019 (COVID-19);tratidional Chinese medicine;diagnosis and treatment program;prevention and treatment program;clinical practice guidelines
Abstract：ObjectiveTo evaluate the clinical efficacy and safety of traditional Chinese medicine Lianhua Qingke granule （LH granule）in the treatment of coronavirus disease 2019（COVID-19）.MethodA randomized controlled method was used in this clinical study. The subjects were confirmed COVID-19 patients over 18 years old with cough symptoms，and the diagnosis is based on Diagnosis and Treatment Protocol for COVID-19 （Trial Version 6）. The subjects were randomized to usual treatment （control group） or in combination with LH granule （1 bag，thrice daily） for 14 days. The primary endpoint was the rate of cough recovery.Result57 subjects were included in this study， including 25 in the control group and 32 in the observation group. The recovery rate of cough in observation group was 90.6%（29/32）， which was significantly higher than 64.0%（16/25）in the control group （P<0.05）. The effective time of cough treatment was significantly earlier than that of the control group （P<0.05）， the median time to cough recovery was markedly shorter in treatment group （P<0.05）. The recovery rate of expectoration in observation group was 94.7% （18/19）， which was significantly higher than 57.1%（8/14）in the control group （P<0.05）. The effective time of expectoration treatment was significantly earlier than that of the control group （P<0.05）， and the median time to expectoration recovery was markedly shorter in treatment group （P<0.05）. The improvement rate of CT in the observation group was 96.9% （31/32）， which was significantly higher than 72% （18/25） in the control group （P<0.05）. After treatment， the oxygenation index of the two groups increased in varying degrees， and the increase rate of the observation group was significantly higher than that of the control group （P<0.05）. There was no significant difference in the recovery rate of fever， fatigue， dry pharynx and sore throat between the observation group and the control group， and it showed that LH had a better improvement trend on the basis of usual treatment.ConclusionOn the basis of routine treatment，LH can significantly improve the symptoms of cough and expectoration of COVID-19 patients， shorten the duration of cough and expectoration symptoms， reduce lung pathological damage， improve respiratory function， and improve the symptoms of fever， fatigue， dry pharynx and sore throat.
Keywords：coronavirus disease 2019 （COVID-19）;Lianhua Qingke granule;cough;randomized and controlled;clinical study
Abstract：ObjectiveTo observe the effect of modified Tongqiao Huoxuetang on circulating blood flow and wall shear stress of vertebrobasilar dolichoectasia (VBD) due to blood stasis and channel blockage.MethodA total of 97 patients admitted in our department from October 2017 to August 2019 were collected. The traditional Chinese medicine (TCM) syndromes were consistent with blood stasis and channel blockage, and diagnosed as VBD by magnetic resonance angiography (MRA). The patients were divided into experimental group (48 cases) and control group (49 cases). Control group was given basic therapy and placebo of TCM, while treatment group was given basic therapy and modified Tongqiao Huoxuetang for 30 days. The degree of relief of vertigo symptoms, vertigo symptom scale (VSS), activity balance confidence (ABC), transcranial doppler (TCD) bilateral vertebral artery and basilar artery blood flow velocity [systolic blood flow velocity (Vs), mean blood flow velocity (Vm), diastolic blood flow velocity (Vd)], mean blood flow differences between (MFV), pulsatility index, resistance index (RI), and wall shear force (WSS) were observed before and after treatment.ResultCompared with control group before treatment, the score of ABC scale in control group after treatment was markedly higher, while the score of VSS was significantly lower (P<0.05). However, there was no statistical significance in the score of vertigo symptom. Compared with treatment group before treatment, the symptom grade of vertigo degree and the score of VSS in treatment group after treatment were substantially lower, while the score of ABC scale was significantly higher (P<0.05). Compared with control group, the score of vertigo degree symptoms and VSS in treatment group were markedly lower, while the score of ABC scale was significantly higher (P<0.05). Compared with control group before treatment, Vm, MFV and WSS of bilateral vertebral artery in control group after treatment were substantially higher, while RI was significantly lower (P<0.05). However, there were no statistical significances in Vs, Vd and PI in control group before and after treatment. Compared with treatment group, Vs, Vd, Vm, MFV and WSS of bilateral vertebral artery in treatment group after treatment were markedly higher, while RI was significantly lower (P<0.05). However, there was no statistical significance in PI of experimental group before and after treatment. Compared with control group after treatment, Vs, Vd, Vm, MFV and WSS of bilateral vertebral artery in treatment group after treatment were substantially higher, while there was no statistical significance in PI and RI. Before and after treatment, there were similar changes in blood flow parameters of the basilar artery and bilateral vertebral artery.ConclusionModified Tongqiao Huoxuetang could improve the clinical symptoms of dizziness or vertigo in patients of VBD due to blood stasis and channel blockage, and the mechanism might be related to the improvement of post-circulation hemodynamics by Tongqiao Huoxuetang.
Keywords：vertebrobasilar dolichoectasia;Tongqiao Huoxuetang;blood stasis and channel blockage;blood flow;wall shear stress
Abstract：ObjectiveTo investigate the protective effect of cerebrospinal fluid containing Tongqiao Huoxuetang (TQHXT) on oxygen-glucose deprivation/reoxygenation (OGD/R)-induced brain microvascular endothelial cells (BMECs), in order to explore the underlying mechanisms.MethodPrimary BMECs were extracted by enzymatic digestion, and the cells were randomly divided into six groups: the normal control group, the OGD/R group, the TQHXT group（20%）, the nimodipine（NMDP） group (10 μmol·L-1), the cabozanix group (1 μmol·L-1) and the combination group. Except for the normal control group, the cells in the other groups were rapidly reoxygenated for 24 h after 2 h of oxygen-glucose deprivation, the OGD/R modeling was performed, and the rats were administered with drugs by groups. BMECs were identified by cell immunofluorescence staining, morphological and ultrastructural changes of OGD/R-induced BMECs were observed, and changes in cell transmembrane resistance (TEER) were detected. The levels of nitric oxide (NO), the activity of lactate dehydrogenase (LDH), the fluorescence intensity of reactive oxygen species (ROS) and the content of tissue-type plasminogen activator (tPA) were measured with kits. Intracellular Ca2+ concentration and cell apoptosis were detected by flow cytometry, and the expression of CD34 was observed. The protein expressions of zonula occluden-1 (ZO-1), vascular endothelial growth factor (VEGF), adhesion kinase (FAK), and Paxillin were detected by Western blot.ResultCompared with the normal control group, the cells in the OGD/R group were shrinking and rounded, TEER value and ZO-1 protein expression in cells were significantly decreased, the contents of NO, LDH and ROS in cells were significantly increased, the content of tPA was significantly decreased, the concentration of Ca2+ and the apoptosis in the cells were significantly increased, CD34 was expressed in cells, and the protein expressions of VEGF, FAK and Paxillin were significantly increased (P<0.01). Compared with the OGD/R group, cell damage in the TQHXT group was significantly improved, the TEER value and ZO-1 protein expression in cells were significantly increased, the contents of NO, LDH and ROS in cells were significantly reduced, the content of tPA was significantly increased, the concentration of Ca2+ and the apoptosis in the cells were significantly reduced, CD34 expression increased in cells, and the protein expressions of VEGF, FAK and Paxillin were significantly increased (P<0.05，P<0.01).ConclusionCSF containing TQHXT protects BMECs from OGD/R injury possibly by promoting angiogenesis through the VEGF-VEGFR2/FAK/Paxillin signaling pathway.
Abstract：ObjectiveTo explore the mechanism of Shaoyaotang in the treatment of ulcerative colitis (UC) based on toll-like receptor 4 (TLR4)/nuclear factor kappaB (NF-κB) signaling pathway.MethodA total of 50 Wistar rats were selected, including half male and half female. The damp-heat UC rat model was replicated by the methods of the combination of diseases and syndromes and the combination of 2, 4, 6-nitrobenzene sulfonic acid (TNBS) and ethanol. After the successful modeling, the model rats were randomly divided into model group, salazulesulfonate group, and low, medium and high-dose Shaoyaotang groups, and 10 rats (half male and half female) were selected as the blank control group. Low, medium and high-dose Shaoyaotang groups were given 6, 12, 24 g·kg-1 bygavage, and salazonyl arsenic group was given 1 g·kg-1 bygavage. Blank control group was given the equal volume of normal saline for 21 consecutive days. Colon samples were collected after the last administration, and the expressions of TLR4, NF-κB p65 and IL-6 mRNA in colon tissues were detected by fluorescent quantitative polymerase chain reaction (Real-time PCR）, and the expressions of TLR4, NF-κB p65 and IL-6 protein in colon tissues were detected by Western blot.ResultCompared with the blank control group, the relative expressions of TLR4, NF-κB p65, IL-6 mRNA and protein in the model group were significantly increased (P<0.05). Compared with the model group, the expression levels of TLR4, NF-κB p65 and IL-6 mRNA and protein in the salazopyridine group and Shaoyaotang groups were significantly decreased (P<0.05).ConclusionShaoyaotang can inhibit the development of UC by regulating the expressions of TLR4, NF-κB p65 and IL-6 mRNA and proteins in the TLR4/NF-κB pathway.
Abstract：Classical famous prescription Dihuang Yinzi is widely used in modern clinical practice，and can treat many kinds of diseases，especially the diseases of nervous system in internal medicine. Its clinical effect is accurate，but it has not been converted into Chinese patent medicine preparations. Therefore，the authors have collected ancient traditional Chinese medicine（TCM） literatures of Dihuang Yinzi by the methods of bibliometrics，and selected and sorted out 254 pieces of effective data， involving 144 ancient books of TCM，and systematically summarized and analyzed the historical development origin，main treatment syndrome，formula making principle，dosage，preparation method，decoction method and medicine taking method of Dihuang Yinzi，in order to provide the ancient literary evidence support for the development and clinical application of classic famous prescriptions. It is found that Dihuang Yinzi was from Xuanming Lunfang written by LIU He-jian，a doctor of Jin dynasty. It was composed of 12 kinds of herbs，namely Rehmanniae Radix Praeparata，Morindae Officinalis Radix，Corni Fructus，Cistanches Herba，Dendrobii Caulis，Aconm Lateralis Radix Praeparaia，Schisandrae Chinensis Fructus，Cinnamomi Ramulus，Poria，maimendong，Acori Calami Rhizoma and Polygalae Radix, and mainly used for the treatment of Yinfei. The later records of Dihuang Yinzi mostly followed the prescription composition and main treatment set forth in Xuanming Lunfang，and its clinical application was expanded. In the 199 articles with the indications for disease treatment，Yinfei was the most commonest indication, and took up about half of the total，which was followed by stroke，taking up about two fifths of the total. It was also used for the treatment of sudden aphonia，flaccidity syndrome，vertigo，enuresis. Dihuang Yinzi has a wide range of treatment，but the pathogenes is always belongs to "the deficiency of water and fire in the kidney". The recipe of Dihuang Yinzi was unique，and can be used to treat both the upper and lower parts of the body，as well as both the outward symptoms and root causes of an illness at the same time， in particular，it mainly focuses on the treatment of the lower and the root. Among the 56 literatures with drug dosage records，about one third of them inherited the records of Xuanming Lunfang: "Equal division，the top is the end，3 qian for each dose." The dosage was generally light. The preparations are mostly decoction and boiled powder. In the decocting and taking methods，it was suggested that "turbid medicine shall be boiled for a short time，and taken after several boilings，with no limit to time."
Keywords：classical famous formulas;Dihuang Yinzi;ancient books of traditional Chinese medicine;textual research;attending diseases;drug dosage;Xuanming Lunfang
Abstract：ObjectiveTo explore the effect and mechanism of esculentoside on lipopolysaccharide-induced mastitis in mice.MethodFemale BALB/c mice were randomly divided into control group, model group, dexamethasone group (DEX, 5.0 mg·kg-1) and esculentoside group (50, 25, 12.5 mg·kg-1). The mastitis model of postpartum female mice (BALB/c) induced by lipopolysaccharide (LPS) was used to analyze the pathological conditions of breast tissue and the activity of recombinant myeloperoxidase（MPO）, factor content and oxidative stress level. Western blot was used to evaluate the effect of esculentoside on Toll-like receptor4 (TLR4)/nuclear transcription factor (NF-κB) signaling pathway proteins.ResultCompared with the normal group, the breast tissue of the model group had typical mastitis changes, such as hyperemia and congestion, the level of MPO increased, and the expression levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) increased significantly (P<0.01). Compared with LPS model group, esculentoside groups could significantly improve the inflammatory damage of mammary gland tissue, reduce the secretion of neutrophils and the activity of MPO, the expression levels of pro-inflammatory factors TNF-α, IL-1β and IL-6 were also significantly down-regulated, the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were increased, while the level of malondialdehyde (MDA) was decreased, and the activation of TLR4/NF-κB signaling pathway was inhibited by esculentoside（P<0.05，P<0.01）.ConclusionEsculentoside have a protective effect on lipopolysaccharide-induced mastitis in mice, which may be related to the inhibition of TLR4/NF-κB signaling pathway protein expression.
Abstract：ObjectiveTo observe the expression of brown adipose tissue (BAT), cells, proteins and corresponding genes in Yang deficiency model mice induced by Rhei Radix et Rhizoma suspension, and to explore the thermogenesis of processed products of Aconiti Lateralis Radix Praeparata with Jianchang faction characteristics.MethodTwenty mice, half male and half female, were randomly selected as the normal female and male groups. And the other 80 mice were administrated with Rhei Radix et Rhizoma suspension (the content of 0.25 g·mL-1) to establish Yang deficiency model, after the model was established, they were randomly divided into the model female and male groups, female and male groups of Shengfupian, female and male groups of Yinfupian, female and male groups of Yangfupian, 10 mice in each group. Mice were intragastric administrated with corresponding medical solution for two weeks (1.54 g·kg-1·d-1) according to groups. Normal group and model group were given equal volume distilled water. After administration, BAT of scapular region of mice was collected and the changes of BAT cells were observed by hematoxylin-eosin (HE) staining. The expression of uncoupling protein 1 (UCP1) and its mRNA were detected by Western blot and real-time fluorescence quantitative polymerase chain reaction (Real-time PCR).ResultCompared with the normal group of the same sex, the proportion of BAT in the model group decreased significantly (P<0.01). Compared with the model group of the same sex, the proportion of BAT in female mice from Shengfupian and Yinfupian groups increased significantly (P<0.01), while there was no significant difference between each administration group and model group in the male mice. Compared with normal mice of the same sex, there were many scattered vacuoles in BAT cells of the model group, and fewer cells could be observed due to larger vacuoles. Compared with the model group of the same sex, BAT cells in mice from the Shengfupian group showed fewer vacuoles, smaller cells and tight arrangement, the density of BAT cells in mice from the Yangfupian group also increased significantly, while the vacuoles in BAT cells of mice from the Yinfupian group decreased relatively and the cells did not increase significantly. Compared with the same sex mice, the expression level of UCP1 in the model group and the normal group was statistically significant (P<0.05, P<0.01). In the female mice, the expression level of UCP1 in Yangfupian group was significantly higher than that in the model group (P<0.05), each administration group of male mice was significantly different from that of the model group of the same sex (P<0.05), of which Yangfupian was the most significant. The relative expression of UCP1 mRNA in the model group was significantly lower than that in the normal group of the same sex (P<0.05, P<0.01). In the female mice, compared with the model group, the relative expression levels of UCP1 mRNA in Yangfupian group, Shengfupian group and Yinfupian group increased significantly (P<0.05, P<0.01), compared with Yangfupian group, the relative expression levels of UCP1 mRNA in Shengfupian and Yinfupian were also significantly different (P<0.05). In the male mice, compared with the model group, the relative expression of UCP1 mRNA in Yangfupian group was significantly increased (P<0.01), but there was no significant difference in Shengfupian group and Yinfupian group, in addition, compared with Yangfupian group, the relative expression of UCP1 mRNA in Shengfupian group and Yinfupian group had significant difference (P<0.05).ConclusionShengfupian, Yinfupian and Yangfupian all have obvious improvement on Yang deficiency syndrome induced by Rhei Radix et Rhizoma suspension. The mechanism may be to promote the expression of UCP1 protein and its mRNA and enhance the activity of BAT. And the effect of Yangfupian is the best.
Abstract：ObjectiveTo establish a mouse model of diminished ovarian reserve （DOR） induced by tripterygium wilfordii polyglycosides （TWP）， and to explore the therapeutic effect of Dingkundan （DKD） on DOR， so as to provide scientific basis for its clinical application.MethodThe 60 female Blab/c mice with regular estrous cycle were randomly divided into blank group， model group， low，medium and high-dose DKD group， DKD group and estradiol valerate group， with 10 mice in each group. Except the blank group， the other groups were given 40 mg·kg-1 TWP suspension. Meantime，low，medium and high-dose DKD group were given 1.64，3.28，6.56 g·kg-1 DKD suspension respectively， and estradiol valerate group was given 0.15 mg·kg-1 estradiol valerate suspension by gastric lavage once a day for 30 days. The general condition， body weight， estrous cycle and gonad index of mice were observed， serum follicle-stimulating hormone （FSH）， luteinizing hormone （LH） and estradiol （E2） were determined by radioimmunoassay， ovarian morphology and follicle count were observed by hematoxylin-eosin staining （HE） staining.ResultCompared with the blank group， most of the mice in model group had disordered estrous cycle， uterine and ovarian indexes decreased （P<0.05）， serum FSH increased （P<0.05）， LH was on an upward trajectory， E2 was on a downward trend， and the number of growth follicles and corpus luteum decreased and the number of atresia follicles increased（P<0.05）. Compared with the model group， half of the mice in DKD group resumed regular estrous cycle， however， the estrous cycles of mice in estradiol valerate group were stagnated during estrous period. In medium-dose， high-dose DKD group and estradiol valerate group， the uterine and ovarian indexes of the mice were increased， the serum FSH value decreased （P<0.05） and serum LH was on a downward trend， high-dose DKD group and estradiol valerate group increased the levels of serum E2 （P<0.05）. In DKD group， the number of growth follicles and corpus luteum were increased and the number of atresia follicles were reduced （P<0.05）， with the best effect at medium dosage. And in estradiol valerate group， the number of primitive follicles， sinusoidal follicles and corpus luteum were increased （P<0.05）， but the number of atresia follicles had no difference to the model group.ConclusionDKD can improve serum sex hormones， promote follicular development and reduce follicular atresia， which can play a therapeutic role in the treatment of DOR.
Abstract：ObjectiveTo study the effect of Schisandrae Chinensis Fructus lignans (SCL) on learning and memory ability of D-galactose（D-gal）-induced aging model mice.MethodICR mice were randomly divided into 4 groups: normal group (distilled water, subcutaneous injection with normal saline), model group (distilled water, subcutaneous injection with 200 mg·kg-1D-gal), piracetam group (oral administration with 200 mg·kg-1 piracetam, subcutaneous injection with 200 mg·kg-1D-gal), low-dose SCL group (oral administration with 50 mg·kg-1·d-1 SCL, subcutaneous injection with 200 mg·kg-1·d-1 D-gal), medium-dose SCL group (oral administration with 100 mg·kg-1·d-1 SCL, subcutaneous injection with 200 mg·kg-1·d-1D-gal), high-dose SCL group (oral administration with 200 mg·kg-1·d-1 SCL, subcutaneous injection with 200 mg·kg-1·d-1 D-gal). The drugs were administered continuously for 10 weeks. Dark test and Morris water maze test were performed to observe the effect of SCL on the learning and memory ability of D-gal-induced aging mice. The activities of superoxide dismutase (SOD) and malondialdehyde (MDA) in mouse brain tissue were detected by chemical colorimetry. The expressions of peroxiredoxin-6（Prdx6） and glutathione peroxidase 1（GSH-Px1） mRNA in mouse brain tissue were detected by polymerase chain reaction (PCR). The expressions of Prdx6 and GSH-Px1 protein in mouse tissues were detected by Western blot.ResultIn behavioral experiments, compared with normal group, the number of dark avoidance errors in model group significantly increased (P<0.05), the latency was significantly reduced (P<0.01), and the number of mouse passes and the target quadrant residence time were significantly reduced (P<0.01), which can be used as an indicator of successful modeling. Compared with the model group, the number of errors in the piracetam group, and medium and high-dose SCL groups was significantly reduced (P<0.05，P<0.01), and the latency was significantly prolonged (P<0.05，P<0.01). At the same time, the number of water maze passes and the target quadrant retention time in the high-dose SCL group increased significantly (P<0.01). The results of biochemical indicators showed that compared with normal group, the SOD activity in brain tissue of model group mice was significantly reduced (P<0.01), while the MDA content was increased (P<0.05). Compared with the model group, SOD activity in the brain tissues of piracetam group, and low, medium and high-dose piracetam groups was significantly increased (P<0.05), whereas the level of MDA was reduced (P<0.05). The expressions of Prdx6 and GSH-Px1 were significantly increased (P<0.05，P<0.01), indicating that the SCL administration group was dose-dependent.ConclusionSCL can improve the learning and memory ability of D-gal-induced aging mice, which may be related to the anti-oxidation ability of SCL and the up-regulation of Prdx6 and GSH-Px1 expressions in mouse brain tissue.
Keywords：Schisandrae Chinensis Fructus lignans;learning and memory ability;glutathione peroxidase 1（GSH-Px1）;peroxiredoxin-6（Prdx6）
Abstract：ObjectiveTo study the interaction between Tanreqing injection and commonly used antibiotics against multi-drug resistant Pseudomonas aeruginosa (MDR-PA) and the effect on bacterial efflux pump.MethodAntibiotic susceptibility test was performed with bacteria. Paper diffusion method (Kirby-Bauer, KB) combined with efflux pump inhibitor (50 mg·L-1) was used to measure the diameter of the inhibition zone, and real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect gene expression of efflux pump Positive efflux pump strain. KB method was used to observe the changes of Tanreqing (final concentration 3 g·L-1) and antibiotics on the diameter of the zone of inhibition. Strains were co-cultured with Tanreqing and antibiotic sub-inhibitory concentrations for Real-time PCR detection. KB method was used to observe the effect of Tanreqing on the diameter of bacteriostatic ring after the continuous use of efflux pump-positive bacteria.ResultTwo MDR-PA efflux pump-positive strains were identified and screened. Tanreqing has synergistic antibacterial effect with aloxicillin, aztreonam, meropenem, ceftazidime, cefoperazone, and Shupushen. In inhibiting the expression levels of bacterial efflux pump genes, the four drugs were compared by the effect: cefoperazone>Tanreqing>ceftazidime>Shupushen. After Tanreqing continued to act on efflux pump-positive strains, it could have a better effect in combination with ceftazidime, cefoperazone, and Shupushen.ConclusionTanreqing, ceftazidime, cefoperazone, and Shupushen can reduce the drug resistance of bacteria by down-regulating the expressions of bacterial efflux pump genes, and reducing the clinical dose of antibiotics, and thus play a bacteriostatic effect.
Abstract：ObjectiveCompare the anti-tumor effect and mechanism of Liushenwan and realgar (As4S4) on human endometrial cancer cells JEC.MethodThe release of Asin Liushenwan and As4S4 was measured by atomic absorption spectrometry. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used for cell proliferation, and cell migration was measured by Transwell assay. Flow cytometry and Western Blot were used to determine apoptosis and DNA damage.ResultThe dissolution of As in Liushenwan was 17.4%, and that of As4S4 was only 1.6% according to atomic absorption assay. With the same content of As, compared with the As4S4 group, the cell viability in the 3，10 mg·L-1 Liushenwan groups was decreased (P<0.05), the early apoptosis rate was significantly increased in 0.25，0.5，1 mg·L-1 Liushenwan groups (P<0.05), the rate of cell migration was decreased in 1 mg·L-1 Liushenwan group (P<0.05), the expressions of cleaved cysteinyl aspartate-specific proteinase-3 (cleaved Caspase-3), cleaved cysteinyl aspartate-specific proteinase-7 (cleaved Caspase-7), cleaved poly ADP-ribose polymerase (cleaved PARP), phosphorylated histone (p-H2AX), phosphorylation of checkpoint kinase 2 (p-CHK2) and ataxia-telangiectasia mutated (ATM) were increased in 1 mg·L-1 Liushenwan group (P<0.05), while the expression of phosphorylation of ataxia-telangiectasia mutated rad3-related (ATR) was decreased in 1 mg·L-1 Liushenwan group (P<0.05), with no significant changes in the expressions of cysteinyl aspartate-specific proteinase-3 (Caspase-3) and cysteinyl aspartate-specific proteinase-7 (Caspase-7).ConclusionWith the same content of As, both Liushenwan and As4S4 could inhibit JEC cell proliferation and migration, and induce cell apoptosis and DNA damage. Liushenwan has a stronger effect than As4S4. It is suggested that there are other components in Liushenwan with an anti-tumor effect in cooperation with As.
Abstract：ObjectiveTo observe the effects of Fushengong prescription on p38 mitogen-activated protein kinase（p38 MAPK） signal pathway of rats with chronic renal failure（CRF），and to explore its mechanism of reducing inflammatory reaction of renal tissues and delaying the progress of renal interstitial fibrosis.MethodThe 55 male Sprague-Dawley rats were randomly divided into normal group，model group， and low，medium and high dose groups of Fushengong prescription，with 11 rats in each group.The normal group was routinely reared， and the other four groups of rats were fed a diet containing 0.5% adenine to produce a model of CRF， which was continuously molded for 21 days.After successful modeling，all rats switched to conventional feed.Normal group and model group were given normal saline 20 mL·kg-1，and each group of Fushengong prescription was given 4，8，16 g·kg-1 of water prescription once a day for 30 days.After the experiment，Masson staining was used to observe the degree of renal interstitial fibrosis.The expression of monocyte chemotactic protein-1（MCP-1） in renal tissues was detected by immunohistochemistry. The expression of phosphorylated p38 mitogen-activated protein kinase（p-p38 MAPK） and transformed growth factor-β1（TGF-β1） in renal tissues were detected by Western blot.ResultCompared with normal group，the renal interstitial collagen deposition increased significantly，the average optical density value of MCP-1 and the expression levels of p-p38 MAPK and TGF-β1 also increased significantly in model group （P<0.05）. Compared with model group，the renal interstitial collagen deposition reduced significantly，the average optical density value of MCP-1 and the protein expression levels of p-p38 MAPK and TGF-β1 also decreased significantly in each dose group of Fushengong prescription（P<0.05）.ConclusionFushengong prescription can effectively inhibit the expression of related inflammatory factors in the renal tissue of CRF rats，so as to reduce the inflammatory response in the renal tissue and delay the progress of renal interstitial fibrosis，the mechanism of which may be related to inhibit the activation of p38 MAPK signal transduction pathway.
Keywords：Fushengong prescription;chronic renal failure;inflammatory reaction of renal tissues;renal interstitial fibrosis;p38 mitogen-activated protein kinase signaling pathway;Liuwei Dihuangwan
Abstract：ObjectiveTo observe the effect of Taohong Siwu granule on dysmenorrhea rats.MethodForty-eight healthy SD female rats were randomly divided into six groups: control group, model group, Taohong Siwu granule groups (18, 9, 4.5 g·kg-1) and Fuke Qianjin tablet group (0.08 g·kg-1). The rats were given Bugaorer (0.35 mg·kg-1) every day and ice-water bath for 8 minutes for 10 consecutive days. Oxytocin was injected intraperitoneally on the 11th day. The writhing reaction of rats was observed, and the effect of Taohong Siwu granules on hemorheology was measured. The levels of 6-ketoprostacyclin F1α(6-keto-PGF1α), thromboxane B2(TXB2), prostaglandinF2α（PGF2α）, PGE2 of rats were detected by enzyme-linked immunosorbent assay (ELISA). Ca2+ in uterine tissues was scanned by laser confocal microscopy.ResultEach dose of Taohong Siwu granule group reduced the number of writhing, the time of writhing and the incidence of writhing in dysmenorrhea rats, which were significantly lower than those in model group (P<0.05，P<0.01). Compared with the model group, high, medium and low-dose Taohong Siwu granules groups could reduce blood viscosity and plasma viscosity, and Taohong Siwu granules can significantly reduce the content of TXB2, Ca2+，PGF2α, and significantly increase the content of 6-keto-PGF1α，PGE2 (P<0.05，P<0.01).ConclusionTaohong Siwu granule can significantly alleviate dysmenorrhea symptoms in rats. The mechanism may be correlated with the contents of 6-keto-PGF1α, TXB2 and PGF2α, PGE2 and Ca2+.
Keywords：Taohong Siwu granule;cold coagulation and blood stasis;dysmenorrhea;rat
Abstract：ObjectiveTo observe the clinical efficacy of modified Danggui Niantongtang combined with Xuanbitang in treating damp-heat type acute gouty arthritis, and investigate its effect on levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-8 (IL-8) and tumor necrosis factor-α (TNF-α).MethodTotally 126 patients were randomly divided into control group and observation group, with 63 cases in each group. In addition to non-medication treatment, patients in control group were given conventional western medicine treatment for 1 week, while patients in observation group were given modified Danggui Niantongtang combined with Xuanbitang for 1 week. The levels of IL-1β，IL-6，IL-8，TNF-α before and after treatment were observed.ResultThe total effective rate was 79.37% in control group and 95.24% in observation group, with significant differences (P<0.05). After treatment, the scores of arthralgia, tenderness, swelling, activity restriction in both groups were significantly decreased (P<0.01), and those in observation group were lower than those in control group (P<0.05). The levels of uric acid (UA), high-sensitivity C-reactive protein (hs-CRP), erythrocyte sedimentation rate (ESR) were significantly decreased (P<0.01) after treatment, and those in observation group were lower than those in the control group (P<0.05). The levels of IL-1β，IL-6，IL-8，TNF-α were significantly decreased (P<0.01) after treatment, and those in the observation group were lower than those in the control group (P<0.05).ConclusionCompared with conventional western medicine treatment, modified Danggui Niantongtang combined with Xuanbitang has a better efficacy in treating damp-heat type acute gouty arthritis and reducing levels of IL-1β，IL-6，IL-8，TNF-α.
Abstract：ObjectiveTo discuss clinical effect of addition and subtraction therapy of Wuhutang combined with Qingjin Jianghuotang to community acquired pneumonia (CAP) in children with syndrome of phlegm heat closing lung, and to study the influence to inflammatory factors.MethodOne hundred and forty patients were randomly divided into control group (69 cases) and observation group (71 cases) by random number table. Patients in two group of chidren got comprehensive symptomatic treatment measures of anti-infection, antipyretic, expectorant, antiasthmatic and respiratory support of inflammatory factors. The control group was treated with Lingyang Qingfei granules.1 g/time,3 time/day. Patients in observation group added addition and subtraction therapy of Wuhutang combined with Qingjin Jianghuotang, 1 dose/day. The courses of treatment in two groups were 7 days. And temperature, time of antipyretic, time of complete antipyretic and rate of complete antipyretic at the 7th day after treatment were recorded. And release time and disappearance time of cough, expectoration, disappearance time of pulmonary rales and treatment failure were also recorded. And before and after treatment, scores of syndrome of phlegm heat closing lung were graded, and levels of serum high sensitive C-reactive protein (hs-CRP), procalcitonin (PCT), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were detected.ResultAnalyzed by rank sum test, effect in observation group was better than that in control group (Z=2.106, P<0.05), and curative effect of traditional Chinese medicine (TCM) syndrome was also better than that in control group (Z=2.119, P<0.05). Time of antipyretic, time of complete antipyretic, release time and disappearance time of cough and expectoration and disappearance time of pulmonary rales were all shorter than those in control group (P<0.01). Rate of complete antipyretic at the 7th day after treatment in observation group was 96.92%（63/65） higher than 82.81%（53/64） in control group (χ2=7.085, P<0.05). Failure rate of treatment was 9.23%（6/65） lower than 23.44%（15/64） in control group (χ2=4.775, P<0.05). And major symptom, physical sign score, minor symptom score, the total score of syndrome of phlegm heat closing lung and levels of hs-CRP, PCT, TNF-α and IL-6 were all lower than those in control group (P<0.01).ConclusionOn the basis of comprehensive anti-infection treatment, addition and subtraction therapy of Wuhutang combined with Qingjin Jianghuotang can control the clinical symptoms, and the advantages of rapid onset, rapid symptom regression, short course of disease can be found, and it can also reduce the inflammatory reaction, control the progress of the disease. The complete antipyretic rate, disease efficacy and TCM syndrome efficacy are better.
Keywords：community acquired pneumonia;child;syndrome of phlegm heat closing lung;Wuhutang;Qingjin Jianghuotang;inflammatory factors
Abstract：ObjectiveTo discuss the clinical efficacy of Yixinshu capsule for viral myocarditis (VMC) with deficiency of Qi and Yin, and to investigate its antioxidant and anti-inflammatory effect.MethodOne hundred and thirty-two patients were randomly divided into control group (66 cases) and observation group (66 cases) by random number table. Patients in two group got comprehensive treatment of Western medicine, i.e. intravenous drip of creatine phosphate injection for 14 days, 1 g/time, 1 time/day. Coenzyme Q10 capsule, 1 grain/time, 3 times/day after meals. Trimetazidine dihydrochloride tablets, 1 tablet/time, 3 times/day during meals. And critically ill patients got intravenous drip of dexamethasone sodium phosphate injection for 14 days, 10-20 mg/time, 1 time/day. The control group took Wenxin granules orally,One bag at a time,3 times/day. Patients in observation group additionally got Yixinshu capsule, 3 grains/time, 3 times/days. The courses of treatment were 8 weeks in both groups. The serum troponin I (cTnI) and creatine phosphokinase (CK-MB) were monitored, and after treatment, the recovery rates of cTnI, CK-MB were recorded. Before and after treatment, the electrocardiogram was observed and the recovery rate after treatment was recorded. Before and after treatment, the scores of deficiency of Qi and Yin were graded, and levels of creatine phosphokinase (CPK), hydroxybutyrate dehydrogenase (HBDH), lactate dehydrogenase (LDH), aspartate aminotransferase (AST), superoxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GSH-Px), interferon-γ (IFN-γ), interleukin-10 (IL-10), IL-17 and IL-35 were detected. Echocardiography, left ventricular ejection fraction (LVEF), cardiac index (CI), and maximum velocity values between early and late diastolic (E/A) were detected.ResultIn the analysis of rank sum test, clinical efficacy in observation group was better than that in control group (Z=2.151, P<0.05). Recovery rates of cTnI, CK-MB and electrocardiogram in observation group were 82.26% (51/62), 90.32% (56/62) and 80.65% (50/62), higher than 65.00% (39/60), 73.33% (44/60) and 63.33% (38/60) in control group (P<0.05). Levels of serum cTnI, CK-MB, CPK, HBDH, LDH, AST, MDA, IFN-γ and IL-17 were lower than those in control group (P<0.01), while levels of LVEF, CI, E/A, SOD, GSH-Px, IL-10 and IL-35 were higher than those in control group (P<0.01).ConclusionOn the basis of comprehensive anti-infection treatment, Yixinshu capsule can additional protect myocardium by anti-inflammatory and antioxidant effect, reduce myocardial enzyme, promote the recovery of ECG and cardiac enzyme, improve cardiac function and improve the effect of clinical treatment.
Keywords：viral myocarditis;subacute phase;deficiency of Qi and Yin;Yixinshu capsule;antioxidant stress;antiinflammatory
Abstract：ObjectiveTo investigate the effective substance of the choleretic effect of Gardeniae Fructus，and analyze the relationships between its choleretic effect and the HPLC fingerprint chromatogram.MethodHPLC method was applied to establish the fingerprint chromatography of 8 batches of Gardeniae Fructus at different harvest periods. The flow，the content of bile acid，bilirubin and cholesterol in bile were tested，and then the principal component analysis was used to comprehensively evaluate the total choleretic effects of Gardeniae Fructus. After the relationships between the relative peak area of the common peaks and the choleretic effects were explored using grey relationship analysis method，the spectrum-effect relationship of Gardeniae Fructus was established.ResultThe order of the contribution of the chemical components to the choleretic effect at the common peaks was as follow（r>0.8）：P9>P14>P26>P4>P30>P6>P1>P10>P5>P24. Among all peaks，the full wavelength scanning results implied that the peaks 9，14 and 4 might be iridoids, and the peaks 26，30 and 24 might be crocins. By comparing with the standard substances，the peak 9 was finally identified as geniposide.ConclusionThe choleretic effect of Gardeniae Fructus may be the results of multiple components and pathways，and the main components in Gardeniae Fructus with the choleretic effect was from geniposide. In conclusion，these results provide a reference for investigating the material basis of choleretic effect of Gardeniae Fructus.
Abstract：ObjectiveTo analyze the main chemical components in the supernatant and precipitate of Sanajon oral liquid, so as to provide basis for establishing its quality standard and precipitation control technology.MethodUPLC-Q-TOF-MSE was used to analyze the chemical components in the supernatant and precipitate of this oral liquid. The analysis was performed on Waters ACQUITY UPLC BEH C18 column (2.1 mm×100 mm, 1.7 μm) with the mobile phase of 0.1% formic acid solution (A) and acetonitrile (B) for gradient elution (0-1 min, 2%B; 1-2 min, 2%-5%B; 2-4 min, 5%-7%B; 4-6 min, 7%-24%B; 6-10 min, 24%-42%B; 10-12 min, 42%-54%B; 12-15 min, 54%-76%B; 15-18 min, 76%-100%B), the flow rate was set to 0.3 mL·min-1, the column temperature was 30 ℃, the injection volume was 2 µL. The mass spectrographic analysis was used with electrospray ionization (ESI), sample MS data was acquired by time-dependent MSE in negative ion mode, the collection range was m/z 50-1 200 (supernatant) and m/z 50-3 000 (precipitate). Then the chemical constituents were identified by the information of retention time, accurate relative molecular mass and secondary mass spectrum fragment.ResultTotally 61 compounds were identified in the supernatant, including tannins, phenolic acids, flavonoids, amino acids, organic acids, fatty acids, etc. Totally 15 compounds were identified in the precipitate, including tannins, phenolic acids, flavonoids, fatty acids, etc.ConclusionThe hydrolyzed tannin of Sanajon oral liquid may be the potential material basis of its precipitate, and its precipitate is likely to be a complex precipitate mainly composed of ellagic acid and tanned red. The established UPLC-Q-TOF-MSE can quickly and comprehensively analyze the chemical composition of Sanajon oral liquid, which can provide a scientific basis for the researches of its material basis, precipitation mechanism and quality control.
Abstract：ObjectiveTo study on the material basis of Sanguisorbae Radix by column chromatography and liquid chromatography-ion trap-time-of-flight mass spectrometry (LCMS-IT-TOF), and analyze the distribution of different components in Sanguisorbae Radix water extract on D101 macroporous resin and polyamide resin.MethodSanguisorbae Radix water extract was separated by D101 macroporous resin and polyamide resin, and LCMS-IT-TOF was used for detection, chromatography separation was achieved on an ACQUITY UPLC HSS T3 column (2.1 mm×100 mm, 1.8 μm) with the mobile phase consisted of water (A) and acetonitrile (B) for gradient elution (0-10 min, 5%-20%B; 10-18 min, 20%-35%B; 18-23 min, 35%-50%B; 23-28 min, 50%-90%B; 28-30 min, 90%B; 30-33 min, 90%-5%B; 33-35 min, 5%B), the flow rate was 0.3 mL·min-1, the column temperature was 30 ℃. Data acquisition was carried out in electrospray ionization (ESI) under the positive and negative ion modes, the scanning range was m/z 100-1 200. According to mass spectrometry data such as accurate molecular mass and fragment information, combined with literature, different chemical components in loading effluents and ethanol eluents of Sanguisorbae Radix water extract were identified. A heat map of the distribution of components in each fraction was drawn by extracting mass spectrum peak intensity data of each sample. The elution rules of various components were compared visually.ResultThe enrichment and separation of D101 macroporous resin and polyamide resin were obvious. Tannins in Sanguisorbae Radix water extract was mainly concentrated in loading effluent of macroporous resin and its water eluent, triterpenoids were mainly distributed in the 90% ethanol eluent of macroporous resin. In the above effluents and eluents, a total of 63 compounds (including isomers) were identified. Among them, 6 compounds, ellagic acid-4-pyranoarabinoside or its isomer, 6-O-galloylnorbergerin, 3-O-galloylnorbergerin, (6-acetyloxy-5,7-dihydroxy-8-methoxy-4-oxochromen-2-yl) acetate, ethyl 2-methyl-5,6-bis (sulfooxy) benzofuran-3- carboxylate were first discovered in Sanguisorbae Radix.ConclusionThe method can quickly and accurately identify the distribution of components in aqueous extract of Sanguisorbae Radix after column chromatography, providing experimental basis for exploring the pharmacodynamic components and mechanism of Sanguisorbae Radix.
Abstract：ObjectiveTo prepare Periplaneta americana thermosensitive hydrogel and investigate its effect on wound healing in diabetic rats.MethodTaking N-isopropylacrylamide (NIPAM) and acrylic acid (AAc) as monomers, thermosensitive poly(NIPAM-co-AAc) [P(NIPAM-co-AAc)] polymeric material was prepared by free radical polymerization, then thermoresponsive copolymer P(NIPAM-co-AAc)-g-HA was synthesized by conjugating P(NIPAM-co-AAc) to hyaluronic acid (HA). The structure and lower critical solution temperature (LCST) of the graft copolymer were characterized by proton nuclear magnetic resonance spectroscopy (1H-NMR) and ultraviolet spectrophotometry (UV). P. americana thermosensitive hydrogel was prepared by dialysis method, and it was characterized by scanning electron microscope (SEM), rotation rheometer and thermogravimetric analyzer to observe section structure, rheological properties and thermal stability. Differential scanning calorimetry, X-ray diffraction and Fourier transform infrared spectroscopy were employed to identify the inclusion of P(NIPAM-co-AAc)-g-HA temperature sensitive material for P. americana extract, and to investigate the effect of P. americana thermosensitive hydrogel on wound healing in diabetic rats, and the rate of wound healing was calculated by Image-Pro Plus 6.0 software. Hematoxylin-eosin (HE) and Masson staining were used to observe the pathological changes of the wounds of rats in each group.ResultP(NIPAM-co-AAc)-g-HA temperature sensitive material was successfully synthesized, its LCST was between 29 ℃ and 31 ℃, it had a dense and uniform porous structure and could uniformly include P. americana extract. Pharmacodynamic studies showed that P. americana thermosensitive hydrogel group had the best effect on promoting wound healing, its infiltration degree of inflammatory cells was significantly reduced, collagen and fibroblasts arranged neatly and compactly, and the density of neovascularization was significantly increased by comparing with the model group.ConclusionP. americana thermosensitive hydrogel can effectively promote wound healing of diabetic rats and overcome the shortage of marketed P. americana liquid preparations, this paper can provide a reference for the development of P. americana extract preparations to promote wound healing in diabetic patients.
Keywords：Periplaneta americana extract;polymer;thermosensitive gel;diabetes;performance characterization;inclusion;wound healing
Abstract：ObjectiveTo explore the synergistic effect of arbuscular mycorrhizal（AM） fungi mixed inoculation on the growth，physiological and biochemical characteristics，root biomass and terpenoid component accumulation of Aucklandia lappa seedlings，so as to provide a reference for the combination and application of the dominant complementary effect mycorrhizal fungi.MethodThe effect of different AM fungi combined with inoculation on the root mycorrhizal infection rate，plant growth，physiological and biochemical characteristics，root biomass，costunolide and dehydrocostus lactone of A. lappa seedlings were determined by pot inoculation at room temperature.ResultIt was found that AM fungi could form good mycorrhizal symbiosis with the roots of A. lappa.The formation of mycorrhizal symbiosis system could increase the chlorophyll content of A. lappa leaves，increase the activities of catalase（CAT），peroxidase （POD），superoxide dismutase （SOD），reduce the content of malondialdehyde（MDA），and promote photosynthesis of A. lappa. Compared with CK group，AM fungus treatment could significantly promote the accumulation of costunolide and dehydrocostus lactone，and the accumulation of its metabolites，costunolide and dehydrocostus lactone，into roots during the symbiotic cultivation of A. lappa seedlings，indirectly improving the quality of medicines and yield of alantolactone.ConclusionInoculation of AM fungi can improve the root mycorrhizal viability，increase the absorption of nutrients and promote the growth of woody incense.The mixed inoculation treatment of S2，S4 and S5 had the best mycorrhizal effect in artificial cultivation，and the growth and medicinal quality of A. lappa were the best，which provided technical support for the application and popularization of A. lappa mycorrhizal biotechnology.
Abstract：ObjectiveTo establish a molecular identification method for Bupleurum chinense seeds based on ribosomal DNA internal transcribed spacer (ITS) sequence, ensuring the species authenticity of the cultivated seeds of B. chinense.MethodA total of 59 seeds samples of B. chinense and its main cultivated species, marketed B. chinense were collected. The effect of different sampling amounts and different water bath conditions on DNA extraction quality of the seeds was investigated, a DNA extraction method for seeds of Bupleurum was established. Their ITS sequences were obtained by polymerase chain reaction (PCR) and bidirectional sequencing. In addition, 34 ITS sequences of main cultivated Bupleurum species, such as B. chinense, B. scorzonerifolium, B. falcatum and B. smithii, were downloaded from GenBank to enrich identification database of B. chinense seeds. The neighbor-joining (NJ) dendrogram were constructed by MEGA-X 10.0.5 software to investigate the the species identification ability of ITS sequences for B. chinense seeds. And DNA barcoding identification of marketed B. chinense seeds was conducted based on BLAST method and NJ dendrogram method.ResultIn total, 59 ITS sequences were obtained. ITS sequences of B. chinense could be divided into six haplotypes, including seven variable sites. The NJ dendrogram indicated that all the haplotypes of B. chinense could form independent branches, which could be distinguished from other cultivated species of Bupleurum in the collected samples, and possessed the ability to identify species of B. chinense seeds. Based on ITS sequence barcoding identification, 3 of the 19 marketed B. chinense seeds were B. falcatum with a counterfeit rate of 15.8%.ConclusionDNA barcoding technology based on ITS sequence can accurately and reliably identify B. chinense seeds and its adulterants, providing reference for the standardization construction of Chinese medicinal materials seeds.
Abstract：ObjectiveTo analyze the main chemical constituents of traditional Chinese medicine compound Bushao Tiaozhi capsules by using UPLC-Q-TOF-MS technology.MethodThe separation was eluted with Waters CORTECS UPLC C18 column（2.1 mm×150 mm，1.6 μm） in a gradient mode， with methanol-0.1% formic acid as the mobile phase. The column temperature was 40 ℃，the flow rate was 0.24 mL·min-1，and the injection volume was 1 μL. The mass spectrometry condition was X500R QTOF mass spectrometry，the positive and negative electrospray ionization （ESI） was adopted for determine the chromatographic effluent，and the main chromatographic peaks were assigned and distinguished by Q-TOF.ResultA total of 53 chemical constituents were identified by reference confirmation，literature comparison，and high mass spectrometry data analysis. The chemical constituent cluster was composed of 21 flavonoids，10 phenolics，5 monoterpene glycosides，7 diterpene lactones and 10 sesquiterpenes. Furthermore，all of the constituents were surveyed and classified according to their medicinal materials derivation. Among them，the 5 flavonoids components（mangiferin，isoquercitrin，typhaneoside，isorhamnetin 3-O-neohesperidoside，tiliroside）were identified in Microctis Folium for the first time.ConclusionThis study shows that UPLC-Q-TOF-MS technology provides a simple，rapid，and accurate method for the identification of chemical constituents in Bushao Tiaozhi capsules. The identified chemical components mostly cover the main constituents of each medicinal material in the formula，so as to provide a new technological method and theoretical foundation for further defining the pharmacological basis and mechanism of action and optimizing the quality control of Bushao Tiaozhi capsules.
Keywords：Bushao Tiaozhi capsules;ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry;flavonoids;phenolics;terpenes
Abstract：ObjectiveTo investigate the targets and possible mechanism of Didangtang in the treatment of bladder cancer.MethodBased on multiple traditional Chinese medicine and disease databases, the network pharmacology was used to screen potential targets, analyze the biological functions of potential targets, and construct a network of "Chinese medicine-target-path-disease". Bioinformatics analysis was applied in population and gene databases, in order to explore the differential expressions of core targets in tissues, distribution in the population and the correlation with prognosis. The in vitro experiment was used to verify the biological function of Didangtang. The underlying mechanism of Didangtang on the candidate target was detected.ResultA total of 21 core target genes and 16 highly enriched pathways were screened out. A functional network of Didangtang was constructed systematically. At the same time, six targets, namely cadherin 1 (CDH1), CAMP responsive element binding protein 1 (CREB1), colony stimulating factor 2 (CSF2), AP-1 transcription factor (JUN), matrix metalloproteinase 2 (MMP2), and prostaglandin-endoperoxide synthase (PTGS2), were differentially expressed in bladder cancer tissues (P<0.05). Furthermore, JUN and MMP2 were also differentially distributed in population (P<0.05). At the same time, the expression level of JUN was correlated with the prognosis of patients with bladder cancer (P<0.05). The in vitro experiment revealed that Didangtang inhibited the proliferation of bladder cancer cells and decreased the expression of candidate target JUN (P<0.01).ConclusionDidangtang has the characteristics of multiple targets and multiple pathways in treatment of bladder cancer. It is initially confirmed that Didangtang can affect the expression of target JUN and inhibit the proliferation of bladder cancer, which lays a good foundation for further studies on mechanism.
Abstract：By summarizing the existing researches on the prevention and treatment of damp-heat syndrome with traditional Chinese medicine（TCM）， this paper digitized the TCM syndromes of damp-heat syndrome， enriched the TCM theories， and provided reference for the prevention and treatment of damp-heat syndrome and provided innovative ideas for the research of damp-heat syndrome. Damp-heat syndrome can cause pathological changes all over the body. It is easy to induce a variety of acute febrile diseases， including ulcerative colitis， chronic gastritis， chronic hepatitis B， chronic kidney disease， irritable bowel syndrome， pneumonia， cough and asthma， which seriously affects the health of the body. Damp-heat syndrome can be divided into syndrome of damp-heat in upper Jiao， syndrome of damp-heat in middle Jiao and syndrome of damp-heat in lower Jiao .At the same time it can be divided into damp-heat obstructing the lung， large intestine damp-heat， damp-heat containing the spleen， liver-gallbladder damp-heat， and bladder damp-heat syndrome ， nephritic damp-heat syndrome by simulating clinical pathogeny according to basic theories of TCM. Heat-clearing and dampness-expelling is the treatment principle of damp-heat syndrome. TCM achieves the purpose of treating damp-heat syndrome by virtue of its anti-inflammation， anti-oxidation， regulation of intestinal flora， lowering blood glucose， lowering blood lipid and enhancing immunity. Based on the two aspects of TCM and prescription， the application， mainly in the syndrome differentiation of triple energizer differentiation， visceral syndrome differentiation is complementary， pharmacological action and mechanism of TCM in prevention and treatment of damp heat syndrome are reviewed in detail， which is a concrete analysis and discussion， in order to provide relevant experience for data integration and system construction of damp heat syndrome， and provide new ideas and help for follow-up clinical drug research and development and optimization innovation of damp heat syndrome. At present， the material basis and relevant prevention and treatment mechanism of dampness-heat syndrome are still not clear in China， so it is necessary to further study and explore its potential biological diagnostic markers， strengthen the research and development of new Chinese medicine preparation against dampness-heat， and elaborate the mechanism of action of Chinese medicine on damp-heat syndrome in multiple ways and directions.
Keywords：damp-heat syndrome;traditional Chinese medicine;formula;action mechanism;advances research
Abstract：Esophageal cancer is one of the malignant tumors with a high morbidity and mortality in China. According to China's latest cancer report released by the National Cancer Center in 2019, the number of people suffering from esophageal cancer reached 246 000 in 2015, and the death toll reached 188 000. How to effectively treat esophageal cancer and improve the survival rate of patients is one of the most urgent problems in the field of medicine. Phosphatidylinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway is one of the most important signaling pathway for regulating cell survival, differentiation and apoptosis in the body. It also plays an important role in the occurrence and mechanism of various cancers. Recent studies have shown that the activation of PI3K/Akt signaling pathway is an important factor in regulating proliferation, apoptosis, cycle arrest, migration and invasion of esophageal cancer cells. The long-term clinical observation found that traditional Chinese medicine has a stable effect in the treatment of esophageal cancer and little side effects, especially in improving the quality of life of cancer patients and prolonging the survival period of patients. At present, it is a research hotspot to intervene this signal pathway with traditional Chinese medicine in the treatment of esophageal cancer, so as to explore its mechanism of action on esophageal cancer. This paper focused on literatures in CNKI and PubMed databases from 2009 to 2019, with PI3K/Akt signaling pathway, esophageal cancer and miRNA as the key words. A total of 226 literatures were retrieved, and 61 literatures relating to traditional Chinese medicine, esophageal cancer, miRNA and PI3K/Akt signaling pathway were sorted out and summarized. This paper reviewed the mechanism of PI3K/Akt signaling pathway in esophageal cancer, the relationship between miRNA and PI3K/Akt signaling pathway and esophageal cancer, and how traditional Chinese medicine can regulate the expressions of relevant proteins in PI3K/Akt signaling pathway to inhibit cell proliferation, affect cell growth cycle, induce cell apoptosis, inducing cell autophagy, inhibit tumor invasion and metastasis, inhibit angiogenesis. Finally, it can improve esophageal cancer to provide theoretical basis and scientific basis for the treatment of esophageal cancer with traditional Chinese medicine.
Keywords：esophageal cancer;phosphatidylinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway;miRNA;traditional Chinese medicine
Abstract：Steroidal saponins are efficacious substances wildly existed in the herbs，and consist of glycosyl and steroid sapogenin. The biosynthesis pathways of steroidal saponins mainly include the cytosolic mevalonate （MVA） pathway and the plastidial methylerythritol 4-phosphate （MEP） pathway，with the MVA pathway as the main pathway. The key enzymes are involved in the biosynthetic pathway, including 3-hydroxy-3-methyl glutaryl coenzyme A reductase（HMGR），1-deoxy-D-xylulose 5-phosphate synthase（DXS），1-deoxy-D-xylulose-5-phosphatereduetoisomerase（DXR），farnesyl pyrophosphate synthase（FPS），squalene synthase（SS），squalene epoxidase（SE），cycloartenol synthase（CAS），cytochrome P450（CYP450），and steroidalglycosyltransferase（SGTase）. In the paper，the biosynthesis roadmap of steroidal saponins was optimized based on previous studies. According to a comprehensive analysis on studies of key enzymes for the past five years, genes, like HMGR，SS，CYP450 and UGT，were studied more，while other genes，like FPS，SE，CAS，were known less. In conclusion， current studies still focus on the primary stage，but lack direct evidence for the roles of key enzymes. This paper would provide a reference and theoretical support for subsequent studies.