Abstract：Endophytic bacteria exist in the host and do not cause obvious damage to the host，which is an important part of the host ecosystem. Endophytic bacteria are a kind of important microbial resources with diverse species and wide distribution. In the process of long-term coevolution between endophytic bacteria and host，many symbiotic relationships，such as antagonism and reciprocity，have been gradually formed，which can directly or indirectly regulate the growth and development of host，assist host to resist diseases and insect pests，environmental stress and other biological processes. Endophytic bacteria can produce a variety of secondary metabolites in the process of symbiosis with the host，which plays an important role in the development of new natural products. In recent years，endophytic bacteria and their secondary metabolites have been widely used in the research and development of new drugs，biological control and other fields. In this paper，the diversity，species identification and secondary metabolites of endophytic bacterial resources are reviewed，and the future development direction is discussed，hoping to provide reference for the further development and utilization of endophytic bacterial resources.
Keywords：endophytic bacteria of plants;endophytic bacteria of fungi;diversity;secondary metabolites
Abstract：ObjectiveTo analyze the changes of soil microbial community structure before and after planting Gastrodia elata in different producing areas，and to investigate the response of soil microorganisms to the planting of G. elata.MethodITS and 16S rDNA high-throughput sequencing technologies were used to detect fungal and bacterial community compositions in the soil，including the soil without planting G. elata（CK1，CK2），the soil around G. elata（GE1，GE2）before harvesting, and the soil around the rhizomorph of Armillaria（AGE1，AGE2） in Dafang, Guizhou and Jinzhai, Anhui respectively.ResultPrincipal component analysis (PCA) showed that the soil microorganisms changed significantly after G. elata planting as compared with the control soil. The sequencing results showed that the planting of G. elata increased the OTUs number of fungi and bacteria. As compared with the control soil，the diversity and abundance of fungal and bacterial communities showed an increase trend after the cultivation of G. elata in soil of Dafang, Guizhou， such changes of fungal communities were not significant, but the abundance of soil bacteria communities increased in Jinzhai, Anhui as compared with the control soil. The abundance of genera Ilyonectria and Nitrospira increased，while genera Russula decreased significantly both in the soil of Guizhou and Anhui. Furthermore，the abundance of Fusarium and Mortierella increased significantly in the soil of Dafang, Guizhou.ConclusionThe soil microorganisms were out of balance after planting of G. elata, and the abundance of pathogenic microorganisms such as Ilyonectria and Fusarium increased，which may be related to the plant diseases and insect pests of G. elata.
Abstract：ObjectiveTo explore the effects of high temperature stress on the growth characteristics of different Armillaria strains，and to provide guidance for screening excellent Armillaria strains with high-temperature resistance.Method14 strains of Armillaria from different G. elata producing areas were used as experimental materials to observe the growth characteristics and conduct phenotypic classification for the strains. rDNA-IGS sequence analysis was used for molecular identification to further determine the genetic relationship of the tested strains.The strain growth rate, biomass，mycelial length and other indicators under the condition of 23 ℃ （CK） and 30 ℃ high temperature stress were recorded.ResultAll the 14 strains of Armillaria had the highest similarity and the closest relationship with Armillaria gallica，but there were significant differences in growth characteristics among different G. elata producing areas. The 14 strains of Armillaria were classified into Ⅳ groups，and the growth status was groupⅠ>group Ⅱ>group Ⅲ>group Ⅳ. After treatment with high temperature stress，the tolerance of each strain to high temperature also showed obvious differences，as shown in the average growth rate of the mycelial was GZ16>SX1>GZ1. The rank of relative mycelial length was GZ16>SX1>GZ3 and the relative biomass was GZ16>SX4>GZ1>HB1>AH2.ConclusionUnder high temperature stress，GZ16 was best in growth rate，relative length of mycelial，relative biomass and growth state，followed by SX1 and GZ1 strains. The results indicate that strains GZ16，SX1 and GZ1 have the strong resistance to high temperature and excellent growth characteristics at normal temperature，so these three strains are suitable to be produced in main G. elata producing areas in China.
Keywords：Armillaria;Gastrodia elata;high temperature stress;growth characteristics
Abstract：ObjectiveTo establish a research platform for obtaining accurate phenotypic spectrum data is a technical difficulty that needs to be resolved in the research of traditional Chinese medicine resources. For example，the traditional phenotypic characterization method of Armillaria rhizomorph is mostly in a form of descriptive text，which is subjective and empirical. There is an urgent need for an objective and accurate method to characterize the phenotype of honey fungus rhizomorph.MethodBased on the image processing software Image J and the root identification plug-in SmartRoot combined with the Synbiosis ProtoCol 3 image analyzer，the growth picture of Armillaria spp. was analyzed，and the length，growth rate，branching situation，and angle of nascent rhizomorph of Armillaria gallica were measured to establish a measurement system for the phenotypic analysis of Armillaria rhizomorph.ResultBased on the method developed in this paper，the growth length，growth rate，number of branches，angle of nascent rhizomorph，and other phenotypic changes can be analyzed in a real-time manner without affecting the growth of Armillaria gallica. Armillaria spp. grew fastest at 9-12 days after generation，and the angle between the nascent rhizomorph and the parent rhizomorph was nearly vertical. This method had a certain correlation with the dry weight of traditional Armillaria biomass phenotypic parameters，with a high value in practical application.ConclusionThis study has established an objective，accurate，fast and real-time phenotypic analysis and measurement system for Armillaria rhizomorph，which expands the scope of application of SmartRoot and can be used for phenotypic analysis of traditional Chinese medicine resources under controlled experimental conditions.
Keywords：SmartRoot;phenotype profile;Armillaria gallica;rhizomorph;growth rate;total length of rhizomorph
Abstract：ObjectiveAs a source of energy for Armillaria mellea and Gastrodia elata，the woods species as fungus material of G.elata are diverse and play an important role in the development of G.elata industry. In order to explore the impact of different woods species on the quality of G. elata，the plant origins and lignocellulose content of the woods，the yield and quality of G. elata per unit area were systemically analyzed through literature research and investigation on production bases.MethodG. elata and its cultivated woods were collected from four main producing areas （Guizhou, Yunnan, Hubei, and Shaanxi），and the the plant origins of the woods were identified by DNA fragments. The content of lignocellulose in the woods was determined by high-performance liquid chromatography and loss-on-ignition method. The content of polysaccharides of G. elata from these 4 areas was determined by Phenol-sulfuric acid method. The yields and polysaccharide content of G. elata cultivated with different woods species were compared and their correlation with the woods was analyzed.ResultThe woods as fungus material of G. elata were diverse in species, and betulaceae was the most widely used species in cultivation of G. elata. There were differences in the composition ratio of lignocellulose in the woods. Nyssaceae had the highest cellulose content，Moraceae had the highest hemicellulose content and Rosaceae had the highest lignin content. Different woods species had certain effects on the yield and polysaccharide content of G. elata. The maximum yield of G. elata was 1 285.51 g and the lowest yield was 379.30 g. The average mass fraction of polysaccharide content was 241.1 mg·g-1，with a range of 87.95-411.2 mg·g-1. The yield and polysaccharide content of G. elata were highly positively correlated with the cellulose content of the woods, and highly negatively correlated with the lignin content.ConclusionDifferent woods have a significant impact on the yield and quality of G. elata. Choosing the appropriate woods species will be beneficial to the absorption of nutrients for A. mellea and the yield increase of G. elata. This study can provide a scientific basis for the selection of woods species during the cultivation of G. elata.
Abstract：Objiective： In the process of microRNA expression analysis by quantitative Real-time polymerase chain reaction（Real-time PCR），the selection of miRNA plays an important role in data standardization.MethodIn this paper，13 Armillaria gallica.Candidate miRNAs were selected for bioinformatics analysis of their precursors，and the PMRD was used to predict similar sequences of their precursors，and the RNAfold was used to predict the secondary structure of the candidate miRNAs and their similar sequences. Real-time PCR was used to detect miRNAs expression in two genotypes of Armillaria gallica（genotype A，genotype B） before and after salt stress，and geNorm，NormFinder and BestKeeper were used to analyze the stability of miRNAs expression.ResultSecondary structure prediction and characterization of 9 candidate miRNA precursors showed that the miRNA predicted belonged to the miR family with typical stem-loop structure and the mature miRNAs were at the 5' or 3' end of the miRNA precursors.geNorm analysis showed that genotype A Armillaria gallica could select Novel-4* and Novel-9 as reference gene，genotype B could select Novel-9 and Novel-16 as its reference gene.NormFinder analysis showed that Novel-9 was stable in both genotype A and B Armillaria gallica.BestKeeper analysis showed that Novel-12* was stable in genotype A Armillaria gallica and Novel-2* was stable in genotype B Armillaria gallica.ConclusionmiRNA Novel-9 is the best stable reference gene，which lays a foundation for further research on the regulation mechanism of miRNA in Armillaria gallica.
Abstract：ObjectiveIsolate and identify Mycena, expand the resources of geminating fungus of Gastrodia elata and optimize the culture conditions of Mycena，in order to provide information and guidance for the production of geminating fungus of G. elata.MethodJuvenile tuber tissue mass transfer separation and purification technology was used for the separation and purification of strains，traditional morphology microscopy was used to isolate the colony mycelia spores morphological characteristics, such as identification，polymerase chain reaction（PCR） amplification rDNA （Ribosomal DNA） internal transcribed spacer（ITS） was used for sequencing analysis and further homology with NCBI database retrieval，MEGA6 software was used to establish Phylogenetic tree by the Maximum likelihood method （MaximumLikelihood，M-L）, so as to classify and identify isolated strains. At the same time，orthogonal test was used to optimize the optimal growth conditions of Mycena.ResultA total of 86 strains were isolated, which belong to 21 species in 12 genera. WMMFJ，SHXG，WMM-21 and MFJ8103 were identified as M. purpureofusca, and ZT01-6 and ZT01-8 were identified as M. cf. purpureofusca. The growth rate of Mycena in wheat bran medium was significantly higher than in PDA medium. The optimal medium composition for the growth of germinating bacteria was 100 g potato，150 g wheat bran and 20 g corn flour，100 g glucose. And 1，3，5-Trihydroxybenzene significantly promoted the growth of WMMFJ，and played a role in promoting the growth of WMM-21 and ZT01-6，and 2-Methoxyphenol promoted the growth of WMMFJ.ConclusionSix strains of Mycena were isolated and identified，four of them are M. purpureofusca，and two of them are M. cf. Purpureofusca. The separation method improved the separation effect of germinating bacteria.
Abstract：Coronavirus disease-2019 (COVID-19) outbreaks were spreading rapidly around the world in early 2020. This disease is within the category of "damp epidemic" and "damp toxin epidemic" in traditional Chinese medicine, with lung and spleen as the lesion focuses, while dampness and toxin as especially prominent properties. Through clinical observation, we found that dampness would often transform into damp heat during the development and evolution of the disease, and the "triple energizer" treatment was an important therapeutic method, eliminating pathogens through purgation and diuresis: for those with damp-heat accumulation toxin diffused to the triple energizer, we could use Ganlu Xiaodudan as primary prescription, which can spread the upper part, smooth the middle part and infiltrate the lower part, so as to provide a way out for the pathogenic factors and regulate Qi movement. For those with damp-heat in Shaoyang channel, we could use Haoqin Qingdantang to clear heat and promote diuresis, harmonize Shaoyang, eliminate damp-heat and epidemic pathogenic factor. For those with pathogen hidden in membrane source dampness trapping and hidden heat, we could use Dayuanyin to eliminate pathogens between interior and exterior parts, and regulate Qi movement. At the same time we shared three typical cases treated in Wuhan, and conducted an in-depth analysis in this study. Clinically, the method of elimination of pathogens through purgation and diuresis is mainly used in the early and middle stage of this disease. When the disease is still located in Qi system, it also can be applied to mild, ordinary patients and severe patients that have accurate syndrome differentiation. In such cases, it can effectively improve symptoms, reduce pathogenic toxin, truncate and reverse the course of disease, give way to pathogen, and avoid disease aggravating.
Keywords：coronavirus disease-2019 (COVID-19);damp heat;Ganlu Xiaodudan;Haoqin Qingdantang;Dayuanyin;case study;method of elimination of pathogens through purgation and diuresis
Abstract：ObjectiveTo analyze the clinical efficacy of Ganlu Xiaodu decoction combined with western medicine in the treatment of common coronavirus disease-2019（COVID-19）.MethodWith the propensity score matching method，the clinical data of patients with common COVID-19 admitted to Wuhan Third Hospital from January 25，2020 to March 18，2020，were screened out. 115 patients with Ganlu Xiaodu decoction combined with conventional therapy were enrolled in the treatment group according to the inclusion and exclusion criteria，and 115 patients treated with conventional therapy were enrolled in the control group. The disappearance rate of main symptoms（fever，cough，expectoration，chest tightness，fatigue and panting），days of syndrome disappearance，effective rate and disappearance rate of other symptoms，laboratory indexes before treatment and after 7 days' treatment，effective rate in computed tomography（CT） of the lungs，rate of conversion to severe illness and total hospitalization time were compared between the two groups.ResultThe baseline data between the two groups showed no statistically significant difference，but were comparable. After 7 days of treatment，the symptom disappearance rate of fever，cough，expectoration，chest tightness，fatigue and panting in the treatment group was significantly higher than that in the control group（P<0.01）.The average number of days to disappearance of main symptoms in treatment group was significantly less than that in the control group（P<0.01）. The effective rate of main symptoms was 93.9%（108/115）in treatment group, significantly higher than 75.7%（87/115，P<0.01）in control group. In the treatment group，the rate of conversion to severe illness was 0, significantly lower than 16.5% (19/115) in the control group (P<0.01). After treatment for 7 days，the white blood cell count，red blood cell count，c-reactive protein，calcitonin，and D-dimer in treatment group were significantly reduced（P<0.05），while lymphocyte count was significantly increased as compared with those before treatment（P<0.01）, the lymphocyte count in control group was also increased significantly in control group（P<0.01），while C-reactive protein，calcitonin and D-dimer were significantly reduced（P<0.05）. As compared with the control group，C-reactive protein and calcitonin levels in the treatment group were significantly down-regulated（P<0.01），and lymphocyte count rose significantly（P<0.05）.The improvement rate of pulmonary CT in the treatment group was 93.9%（108/115），which was significantly higher than 75.7%（87/115）in the control group（P<0.001）. After treatment，the patients in both groups were cured and discharged，and the average length of stay in the treatment group was（14.70±2.76） d，significantly less than（15.80±4.37） d in the control group （P<0.05）. No adverse events occurred in the treatment group but 6 cases occurred in the control group.ConclusionGanlu Xiaodu decoction combined with western medicine can significantly improve the clinical symptoms of common COVID-19 patients such as fever，cough，expectoration，chest tightness，fatigue, panting and shortness of breath，alleviate the main symptoms effectively，reduce the rate of conversion from common to serious COVID-19，regulate the expressions of relevant peripheral inflammation factors，promote absorption of lung inflammation，shorten hospitalization time，and improve the cure rate，indicating that Ganlu Xiaodu decoction can be effective in the treatment of patients with common COVID-19.
Abstract：Coronavirus disease-2019 (COVID-19) belongs to the epidemic diseases of traditional Chinese medicine (TCM), which is infected by the air with disease and the initial stage of the disease is in upper energizer. TCM holds that the nose is the orifice of the lung and the gateway of the breath. WU Shang-xian, the famous external doctor in Qing dynasty, discussed in Liyue Pianwen that "for the disease in upper energizer, the most effective method is to use the medicine powder via nasal administration and sneeze to disperse". For thousands of years, ancient Chinese medical practitioners had explored and developed the TCM nasal administration method in the struggle against epidemics. Qingkailing is the basic formula for heat-clearing and detoxifying, and researches have clarified its therapeutic effect on upper respiratory tract infections. Therefore, based on TCM nasal administration, this article took Qingkailing as an example to study the feasibility of its nasal preparations for the treatment of COVID-19. On the one hand, it is helpful for the rapid development of Qingkailing nasal preparations for COVID-19. On the other hand, it can broaden the new thinking of TCM in treating epidemic diseases, and give full play to the advantages of TCM in treating epidemic diseases.
Keywords：coronavirus disease-2019 (COVID-19);nasal administration;nasal preparations;Qingkailing;severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2);lung opening at nose;traditional Chinese medicine (TCM)
Abstract：Professor XUE Bo-shou，a master of traditional Chinese medicine（TCM），believes that coronavirus disease-2019 （COVID-19） belongs to the category of "cold epidemic" or "cold-dampness epidemic" in TCM theory. Cold-dampness obstructing the lungs is the key pathogenesis in the early stage of the disease. Exogenous pathogenic factors of cold and dampness are converted into heat in the interior. Over time，the pathogenesis is the disturbance in ascending and descending of Qi movement，loss of vital Qi， deficiency of both Qi and Yin，and even the occurrence of inner blocking causing Yang collapse. Professor XUE believes that the clinical syndrome differentiation and treatment shall be based on six meridians， and the efficacy could be improved by reference to classical prescriptions of plague in the past dynasties. To treat the disease in the early stage， the prescriptions emphasize on diffusing the lungs，inducing resuscitation and expelling pathogenic factors. He advocates the proper use of Ephedrae Herba， which can induce resuscitation of the lungs，diffuse the lungs and expel pathogenic factors，dissipate the interstitial depression and pathogenic factors in the lung interstitium. Therefore，it is considered that Ephedrae Herba is an effective medicine for diffusing the lungs，expelling pathogenic factors，inducing diuresis，eliminating dampness，promoting blood circulation and removing stasis due to cold coagulation. At the same time，the weakness of vital Qi is the root cause of the disease. The prosperity and decline of vital Qi plays a decisive role in the development， prognosis and efficacy of the disease. Professor XUE attaches importance to overall treatment，and emphasize the basis of pathogenesis， vital Qi， and stomach Qi. We shall expel pathogenic factors of disease，while protecting the vital Qi and stomach Qi. This article summarizes Professor XUE's diagnosis and treatment of COVID-19， enumerates and analyzes professor XUE's consultation cases of COVID-19 in designated hospitals in Linyi， Xinjiang and Beijing， in order to provide more ideas for the anti-epidemic effect of TCM.
Abstract：Myocardial injury from coronavirus disease-2019 (COVID-19) is a clinical sign after the novel coronavirus infection, which can be seen in common type and severe type in acute stage, or after recovery of COVID-19. From the mouth and nose, the epidemic and pathogenic factors enter the lung, involving the heart, so that Qi and blood are blocked and developed into myocardial injury. More than 100 patients with COVID-19 and rehabilitation were treated, and we found that some patients infected with novel coronavirus had palpitation, shortness of breath, chest pain, chest oppression, fatigue and other symptoms. Electrocardiogram(ECG) showed myocardial ischemia injury and increased myocardial enzymes, due to the pathological changes of warm pathogen, first invading the lung and reversely spreading to the heart. Myocardial injury due to infection of novel coronavirus, on the one hand, made the patients weak and conditions lingering and hard to heal after COVID-19 was recovered in discharged patients. On the other hand, myocardial injury in severe cases could easily aggravate the disease and even threaten life. According to the different stages of the disease, the severity of the disease, and the patients' physique in the recovery period, different treatment methods were adopted. For the myocardial injury in patients with acute severe COVID-19, Shengmaiyin and Emergency Huiyangtang can be taken， if the poisonous heat disturbs the mind, we can use Angong Niuhuangwan and Zhibaodan as appropriate. For myocardial injury occurred in common type of COVID-19, Zhuye Shigaotang and Shenxiantang can be used. For the pathological changes mainly including myocardial injury after clinical recovery of COVID-19, we can use Shengxiantang, Yangxintang, Chaixiantang and so on. Methods of detoxification, tonifying Qi and Yin, invigorating Qi and ascending Qi collapse, blood circulation and phlegm resolving were used for treatment of myocardial injury from COVID-19, which can not only effectively improve clinical symptoms, but also restore creatine kinase isoenzyme-MB(CPK-MB) and ECG levels to obtain satisfactory results. All of these could reflect that TCM has a notable advantage in the prevention and treatment of this disease.
Keywords：novel coronavirus;coronavirus disease-2019 (COVID-19);myocardial injury;Chinese medicine treatment
Abstract：Traditional Chinese medicine (TCM) plays an important role in the fight against coronavirus diseasef-2019 (COVID-19) epidemic. A total of 241 Chinese patent medicines and 242 prescriptions were recommended by the state and 26 provinces (autonomous regions and municipalities directly under the central government) for the prevention and treatment of COVID-19. A total of 53 varieties of commonly used medicinal materials were selected by analysis, of which 20 were common key varieties in medicinal materials of Chinese patent medicines and prescriptions, including Glycyrrhizae Radix et Rhizoma, Forsythiae Fructus, Pogostemonis Herba, etc. At the same time, some cold-quilt medicines and ethnic medicines also played an important role in the epidemic. By evaluating the supply and regeneration ability of the key varieties of TCM resources under the current COVID-19 epidemic situation, means of the source, regeneration cycle, medicinal parts, new production and market information of 53 kinds of TCM at present, it is suggested that the artificial cultivation of Glycyrrhizae Radix et Rhizoma and Paeoniae Radix Rubra can be strengthened, the development of Lonicerae Japonicae Flos, Isatidis Radix and other domestic medicinal materials can be controlled. In response to the change in market demand, the production variety structure and planting area of Chinese medicinal materials were timely adjusted to improve the quality standard and safety index of Chinese medicinal materials, and in order to provide ideas for the rational allocation of TCM resources and the development of Chinese medicinal materials industry under the epidemic situation.
Keywords：coronavirus disease-2019 (COVID-19);traditional Chinese medicine (TCM);market demand;balance of supply and demand;regenerative capacity;resource allocation;medicinal material industry
Abstract：Patients with low immune function are prone to novel coronavirus infection, which is consistent with the traditional Chinese medicine (TCM) concept of deficiency of vital Qi and invasion of toxin. At present, it is necessary to focus on the development of antiviral drugs, but it is also urgent to study the preparation for regulating the immune system. Mucosal tissue is an important barrier of human immune system. It has an independent immune system with unique functions and structures. It is the body's first line of defense against infection, and is in direct contact with external antigens (such as food, symbiotic bacteria, viruses, etc.). In the resistance to viruses and infections, the mucosal immune system (such as respiratory mucosa, intestinal mucosa, etc.) plays an extremely important role, which can eliminate foreign pathogenic microorganisms or other foreign antigens, so that the virus does not invade the body tissue and cause damage to the body. There are more and more reports on the therapeutic effects of TCM through the mucosal immune system. This paper aims to explore the relationship between mucosal immunity and coronavirus disease-2019 (COVID-19) and the intervention mechanism of TCM, so as to provide useful research methods and therapeutic ideas for the prevention and treatment of COVID-19.
Abstract：The pathological anatomical results of coronavirus disease-2019 (COVID-19) patients showed that excessive inflammatory reaction in the lungs is one of the important causes for such complications as acute lung injury or acute respiratory distress syndrome. Therefore, regulation of immune response may be an effective measure for COVID-19. Alveolar macrophages have a high heterogeneity and plasticity. The dynamic changes of subsets balance and function of M1/M2 alveolar macrophages have a significant effect on pulmonary inflammatory response during the early and late stages of infection. This paper reviews the classification and function of macrophages and explores the mechanism of alveolar macrophage in the pathological process of COVID-19 at different stages and the pharmacodynamic mechanism of traditional Chinese medicine. Besides, it provides ideas for the treatment of COVID-19 with traditional Chinese medicine and other drugs' research and development based on the regulation of macrophage polarization.
Keywords：coronavirus disease-2019;alveolar macrophages;immune response;acute lung injury;traditional Chinese medicine
Abstract：ObjectiveTo observe the effect of Shenling Baizhusan on the protein and mRNA expression of inhibitor of nuclear factor kappa B kinase (IκK)/inhibitor of nuclear factor kappa B(IκB)/nuclear factor kappa B(NF-κB) signaling pathway in the colon of rats with ulcerative colitis (UC) of spleen deficiency and dampness stagnation type, and to explore the mechanism of Shenling Baizhusan in the treatment of UC.MethodThe 48 Wistar rats were randomly divided into normal group, model group, Shenling Baizhusan group (15.6 g·kg-1) and osalazine sodium group (0.68 g·kg-1), 12 rats in each group. The model of UC with spleen deficiency and dampness stagnation was reproduced by trinitrobenzene sulfonic acid (TNBS)/ethanol enema combined with environment and diet intervention.Serum levels of tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β(IL-1β) were determined by enzyme-linked immunosorbent assay（ELISA）. The expression of NF-κB p65, IκBα, IκKβ protein in colon tissue was measured by Western blot and immunohistochemical method, and the mRNA expression of NF-κB p65, IκBα and IκKβ in colon tissue of rats in each group was detected and compared by real time polymerase chain reaction (Real-time PCR).ResultCompared with normal group,the levels of TNF-α,IL-6 and IL-1β in the serum, the protein and mRNA expression of NF-κB p65, IκKβ in colon tissue of the model group was significantly higher than that of normal group (P<0.01), and the protein and mRNA expression of IκBα was significantly lower than that of normal group (P<0.01). Compared with model group,the protein and mRNA expression of NF-κB p65, IκKβ in colon tissue of the Shenling Baizhusan group and osalazine sodium group were significantly decreased (P<0.01), and the protein and mRNA expression of IκBα was significantly increased (P<0.01).ConclusionShenling Baizhusan can obviously down regulate the protein and mRNA expression of NF-κB p65, IκKβ,up regulate the expression of IκBα in colon tissue of UC rats with spleen deficiency and dampness stagnation. The inhibition of IκK/IκB/NF-κB signal pathway activation by Shenling Baizhusan is an important mechanism of its role in protecting intestinal mucosa.
Abstract：ObjectiveTo investigate the effect of maternal intervention of Zuoguiwan on the improvement of serum and pancreatic index in rats of gestational diabetes mellitus(GDM).MethodThe GDM pregnant rats were replicated by monogamous cage and introneal injection of streptozotocin (STZ). The pregnant rats were randomly divided into normal group, model group, insulin group (20 U·kg-1), and low, medium and high-dose Zuoguiwan groups (2.5，5.0，10.0 g·kg-1). The insulin group was given hypodermic injection of portal winter insulin, all drug groups were respectively administrated with corrsponding drugs for 2 weeks, once a day. Meanwhile, control group and model group were given physiological saline by gavage. After that, the levels of serum insulin, fasting plasma glucose（FBG）, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and IL-8 were measured by enzyme-linked immunosorbent assay (ELISA), protein and gene expression levels of Pancreas-duodenal homologous box factor-1 (PDX-1) were respectively detected by Western blot and real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）.ResultCompared with normal group, FBG, TNF-α and IL-6 levels of pregnant rats in model group increased significantly (P<0.01), the level of IL-8 increased uniformly (P<0.05), and serum insulin level, protein and gene expression levels of PDX-1 decreased significantly (P<0.01). Compared with model group, FBG and serum IL-8 levels of pregnant rats in insulin group were significantly lower (P<0.01), while serum insulin level, protein and gene expression levels of PDX-1 increased significantly(P<0.01). FBG, IL-8, IL-6 and TNF-α levels in serum of pregnant rats in medium-dose Zuoguiwan group decreased obviously, meanwhile serum insulin level, protein and gene expression levels of PDX-1 increased significantly (P<0.01).ConclusionZuoguiwan may promote the secretion of insulin by reducing the serum inflammatory factors in GDM rats, and at the same time up-regulate the expression of PDX-1 protein and gene, so as to restrain insulin resistance, reduce the damage of pancreatic cells and improve the blood glucose of rats.
Abstract：ObjectiveTo explore the efficacy and molecular mechanism of Zhizhuwan decoction and its ingredient-contained serums on the proliferation and apoptosis of rat colon interstitial cells of cajal (ICC), and make a molecule-level analysis of the possible mechanism of traditional Chinese medicine (TCM) purgation-tonifying therapy in treating slow transit constipation (STC).MethodA total of 40 rats were divided into Atractylodis Macrocephalae Rhizoma(AMR) group, Aurantii Fructus Immaturus（AFI） group, Zhizhuwan group and blank serum group on random basis, with 10 in each group. Baizhu group was given 17.7 g·kg-1·d-1 of AMR decoction by gavage, AFI group was given 8.9 g·kg-1·d-1 AFI decoction by gavage, Zhizhuwan group was given 26.4 g·kg-1·d-1 Zhizhuwan decoction by gavage, and blank serum group was given 3 mL sterile distilled water for 7 consecutive days, once a day. Drug-contained serums and blank serum were collected from blood of the above groups and diluted to 5%, 10%, 15% and 20% concentrations. Each concentration was intervened for 24 h and 48 h, and the amount and status of ICC were observed. The best intervening concentration and time for each group with cell counting kit-8 (CCK-8) were determined. Rat colon ICC was divided into blank control group, blank serum group, AMR group, AFI group and Zhizhuwan group. ICC proliferation for each group was detected with EdU, ICC apoptosis for each group was detected by flow cytometry, and expressions of X-linked inhibitor of apoptosis protein (XIAP) and proliferating cell nuclear antigen (PCNA) were detected by Western blot.ResultCompared with the normal group, the best intervention concentration for blank serum group, AMR group and AFI group was 10%, while that for Zhizhuwan group was 5%. The best intervention times for the above groups were all 24 h. No distinct difference between the effect of blank control group and blank serum group on the proliferation and apoptosis of ICC was observed. In comparison with blank control group and blank serum group, AMR group, AFI group and Zhizhuwan group showed significant changes in ICC proliferation rate (P<0.05，P<0.01). There was a greater increase in ICC proliferation rate of Zhizhuwan group than that of AMR group and Zhizhu group (P<0.05，P<0.01), with no distinct difference between the changes of ICC proliferation rates in AMR group and AFI group. There was no significant difference between the changes of ICC apoptosis rates in AMR group, AFI group and Zhizhuwan group than in blank control group and blank serum group. There were significant increases in the expressions of XIAP and PCNA in AMR group, AFI group and Zhizhuwan group than in blank control group and blank serum group (P<0.05，P<0.01)， but with little difference among the three groups.ConclusionAt certain concentrations, Zhizhuwan, AFI and AMR all have the effect in improving ICC proliferation by increasing XIAP and PCNA expressions, with no evident effect on the apoptosis of ICC, based on TCM purgation-tonifying therapy, Zhizhuwan has the effect in improving ICC proliferation, with a better effect than single administration with AFI or AMR.
Keywords：Zhizhuwan;interstitial cells of Cajal;proliferation;apoptosis;purgation-tonifying therapy
Abstract：ObjectiveTo explore the effect of different effective parts of Taohe Chengqitang on the synthesis and degradation of extracellular matrix in human kideny-2（HK-2） cells induced by transforming growth factor-β1（TGF-β1）.MethodPetroleum ether extract, ethyl acetate extract, n-butanol extract, raffinate and polysaccharide extract, mirabilite extract were extracted with 70% ethanol by systematic solvent method. The HK-2 cell fibrosis model induced by TGF-β1 was built to intervene the cells in different parts of Taohe Chengqitang with different concentrations (0, 50, 100, 200, 400, 800 mg·L-1). Enzyme-linked immunosorbent assay（ELISA）kit assay was used to detect collagen（Col）-Ⅰα1 and fibronectin （FN）in supernatant to screen out the main active parts. Cell counting kit-8 （CCK-8）method was used to determine the best concentration of intervention site of bioactive components. Western blot analysis was used to detect the expression levels of Col-Ⅰ, Col-Ⅲ, matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase inhibitor2 (TIMP2), and connective tissue growth factor (CTGF). Immunofluorescence assay was used to detect the expression of α-smooth muscle actin（α-SMA）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） analysis was used to detect the mRNA expression of plasminogen activator inhibitor-1（PAI-1）.ResultELISA kit assay demonstrated that compared with the model group, ethyl acetate extract, n-butanol extract and chloroform extract significantly reduced the Col-Ⅰα1 and FN content at the concentrations of 200 and 400 mg·L-1 (P<0.05, P<0.01). CCK-8 assay showed that the cells viability was significantly inhibited with drug intervention at the concentrations of 400 and 800 mg·L-1 (P<0.01). Western blot demonstrated that compared with the model group, ethyl acetate extract, n-butanol extract and chloroform extract decreased the expression levels of Col-Ⅰ, Col-Ⅲ, TIMP2 and CTGF in HK-2 cells induced by TGF-β1, and increased the expression of MMP-2 (P<0.05), with more significant effect in n-butanol extract (P<0.01). The results of immunofluorescence showed that ethyl acetate extract, n-butanol extract and chloroform extract could inhibit the expression of α-SMA (P<0.05), with more significant effect in n-butanol extract (P<0.01). The results of Real-time PCR showed that ethyl acetate extract and chloroform extract inhibited mRNA expression of PAI-1 (P<0.05), with more significant effect in n-butanol extract (P<0.01).ConclusionThe extracts of ethyl acetate, n-butanol and chloroform are the active parts of Taohe Chengqitang with the anti-renal fibrosis effect, with n-butanol extract as the most active part. The mechanism on anti-renal fibrosis may be related to its regulation of extracellular matrix (ECM) synthesis and degradation.
Abstract：ObjectiveTo investigate the effect of salvianolate lyophilized injection and Xueshuantong injection (lyophilized) on the permeability of blood-brain barrier (BBB) via inhibition of metallomatrix protease（MMPs） in cerebral ischemia/reperfusion injury rats.MethodThe focal cerebral ischemia/reperfusion model in rats was built by middle cerebral artery occlusion/reperfusion (MCAO/R) technique. Male Wistar rats were randomly divided into sham operation group, ischemia/reperfusion (I/R) group, edaravone (Eda, 6 mg·kg-1) group, salvianolate lyophilized injection (SLI, 21 mg·kg-1) group, Xueshuantong (XST, 100 mg·kg-1) group and SLI combined with XST (SLI+XST, 21 mg·kg-1+100 mg·kg-1) group. Drugs were injected via tail vein for 2 d, while sham group and I/R group were injected with the same amount of normal saline. Neurological deficit score, hematoxylin-eosin (HE) staining and Nissl staining were assessed 2 d after MCAO/R. The permeability of BBB was observed by the leakage of IgG/CD31. The expressions of Claudin-5，Occludin，collagen-Ⅳ（Col- Ⅳ），Laminin，Fibronectin were observed by immunofluorescence staining，and MMP-2 and MMP-9 were observed by Western blot.ResultCompared with the I/R group, SLI group, XST group and SLI+XST group showed improvements in neurological deficit score, HE staining and Nissl staining. The leakage of IgG was alleviated; The positive expressions of Claudin-5，Occludin，Col-Ⅳ，Laminin，Fibronectin in ischemic penumbra were significantly up-regulated, while the expressions of MMP-2 and MMP-9 were down-regulated. The effect in improving SLI combined with XST was much better than a single factor.ConclusionSalvianolate lyophilized injection and Xueshuantong injection (lyophilized) can alleviate cerebral ischemia/reperfusion injury and exert the synergistic effect when they are used in combination. The mechanisms might be associated with the improvement in the permeability of blood-brain barrier by inhibiting MMPs in cerebral ischemia/reperfusion injury rats.
Abstract：ObjectiveTo explore the effect of anemarrhena asphodeloside BⅡ (TBⅡ) on the expressions of nuclear transcription factor-κB receptor activator factor ligand （RANKL）, RANK and C-FOS genes during osteoclast differentiation.MethodMolecular docking software LeDock was used to score the docking of TBⅡ with RANKL, RANK and C-FOS. RAW264.7 was treated with soluble RANKL（sRANKL） and divided into control group, sRANKL group (model group), Icariin (Ica) group, low-dose TBIⅡ group (2 μmol·L-1), medium-dose TBⅡ group (4 μmol·L-1), and high-dose TBⅡ group (8 μmol·L-1). The corresponding kit was used to detect iconic enzyme (TRAP) of osteoclast differentiation. Total RNA was extracted by trizol method, Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the expressions of C-FOS, upstream RANKL/RANK and downstream nuclear factor of activated T-cells cytoplasmic 1 (NFATC1), and osteoprotegerin OPG.ResultThe molecular docking score were -11.86, -11.38, -12.34 kcal·mol-1, and there might be multiple binding sites between TBII as well as RANKL, RANK and C-FOS. Compared with the control group, the content of TRAP in model group increased significantly (P<0.01), and compared with model group, the content of TRAP in each administration group decreased significantly (P<0.01), and TBⅡ decreased the content of TRAP in a dose-dependent manner. Compared with the control group, the expressions of RANKL, RANK, C-FOS and NFATC1 increased (P<0.01), whereas the expression of OPG decreased (P<0.01) in model group. Compared with model group, the expressions of RANKL, RANK, C-FOS and NFATC1 decreased (P<0.01), while the expression of OPG increased (P<0.01) in each administration group.ConclusionTBⅡ may inhibit the differentiation of osteoclast precursors into osteoclasts, inhibit osteoclast activity, reduce bone resorption and improve osteoporosis by regulating RANKL/RANK/C-FOS signal pathway.
Abstract：ObjectiveTo observe the effects of Shenma Yizhi prescription（SMYZ）on learning and memory， inflammatory factors and oxidative stress related indexes of rats with chronic cerebral ischemia inducing vascular dementia caused by bilateral common carotid artery ligation.MethodThe chronic cerebral ischemia model of SPF grade SD rats was prepared by bilateral common carotid artery ligation. After successful surgery， the rats with cognitive impairment were randomly divided into 5 groups，model， donepezil（0.45 mg·kg-1）， SMYZ-low， medium and high dose（2.97，5.94，11.88 g·kg-1）， with 10 rats in each group， and 10 rats in sham operation group were established. Intragastric administration were given for 8 weeks respectively then followed Morris water maze test. The pathomorphological changes of hippocampus CA1 were observed. The serum levels of acetylcholine(Ach)and acetylcholinesterase (AChE)were detected， and the levels of inflammatory cytokines interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in the serum of rats were detected. The contents of glutathione (GSH)， malondialdehyde (MDA)， superoxide dismutase (SOD) and glutathione peroxidase reductase (GSH-Px) were observed.ResultCompared with sham operation group， the escape latency increased significantly in the model group， and the frequency of platform search， the frequency of crossing the original platform and the percentage of swimming time in the original platform quadrant decreased significantly （P<0.05，P<0.01）. The contents of Ach and the contents of GSH and GSH-Px were significantly decreased （P<0.05，P<0.01）. The contents of AChE， IL-1β and TNF-α and the contents of MDA were significantly increased （P<0.05，P<0.01）. Compared with model group， the escape latency of rats in the in the low， medium and high dose groups of SMYZ prescription was significantly reduced （P<0.05，P<0.01）.The frequency of looking for a platform， the frequency of crossing the original platform and the percentage of swimming time in the original platform quadrant of the rats in the low， medium and high dose groups of SMYZ prescription increased significantly （P<0.05，P<0.01）. The content of AChE in high dose group were significantly reduced（P<0.01）， and the content of Ach in high dose group were significantly increased（P<0.05）， the rat serum IL-1β， TNF-α levels and MDA content were significantly reduced（P<0.05，P<0.01）. The GSH， SOD and GSH-Px levels were significantly increased （P<0.05，P<0.01）.ConclusionSMYZ can improve the learning and memory ability of rats with vascular cognitive disorder caused by chronic cerebral ischemia caused by ligation of bilateral common carotid artery， bilateral common carotid artery ligation， improve the pathological morphology of hippocampal tissue， inhibit inflammatory response， improve cholinergic level and oxidative stress state.
Abstract：ObjectiveTo explore the material basis and mechanism of acute stroke treated with Rhei Radix et Rhizoma based on Homotherapy for Heteropathy using the analysis of proteomics and bioinformatics.MethodA total of 60 male Sprague-Dawley（SD）rats were randomly divided into ischemic stroke（IS） sham-operation group (Sham1), IS model group (IS), IS+ Rhei Radix et Rhizoma treatment group (DH1)，ICH sham-operation group (Sham2), intracerebral hemorrhage（ICH） model group (ICH), and ICH + Rhei Radix et Rhizoma treatment group (DH2), with 10 rats in each group. After cerebral perfusion, the brain tissues were quantified by proteomic analysis, and differentially expressed proteins (DEPs) were identified. Specimens of IS, Sham1, and DH1 groups were collected at 24 hours, while those of ICH, Sham2, and DH2 groups were collected at 48 hours. The common DEPs were analyzed by bioinformatics, and the relevant DEPs were verified by Western blot.ResultRhei Radix et Rhizoma regulated 21 common DEPs associated with acute stroke (including 12 up-regulated and 9 down-regulated). According to Kyoto Encyclopedia of Genes and Genomes（KEGG） analysis, amyotrophic lateral sclerosis (ALS) pathway was enriched, and three proteins [Neurofilament light polypeptide (Nefl), Neurofilament medium polypeptide (Nefm), Neurofilament heavy polypeptide (Nefh)] involved in this pathway. Energy metabolism, ion homeostasis, regulation of synaptophysin, cell cycle and neurogenesis were the common mechanisms of "Homotherapy for Heteropathy". After treatment with Rhei Radix et Rhizoma, the expression levels of GTP binding protein REM2 (Rem2), tyrosine 3-monooxygena (Th), Nefl and neuromodulin (Gap43) were significantly higher than those of the corresponding model group (P<0.05). The expression of Nefl was down-regulated, while the expressions of Rem2，Th and Gap43 were up-regulated, which was consistent with the results of proteomics.ConclusionRhei Radix et Rhizoma-homotherapy-differential protein expression profile is established is study. Energy metabolism, ion homeostasis, regulation of synaptophysin, cell cycle and neurogenesis are the common mechanisms.
Keywords：Rhei Radix et Rhizoma;stroke;homotherapy for heteropathy;proteomics
Abstract：ObjectiveTo investigate that the effect of ethanol extracts from Sophorae Tonkinensis Radix et Rhizoma on the expression of transforming growth factor-β1 (TGF-β1)and Smad3 in the hypertrophic scars of rabbit ears and elucidate its mechanism to improve hypertrophic scars.MethodThe model of hypertrophic ear scar model was established by damaging the inner skin of ears in New Zealand white rabbits.The 49 rabbits were randomly divided into control group, model group, low, medium and high-dose ethanol extracts groups from Sophorae Tonkinensis Radix et Rhizoma (0.4，1.0，2.0 g·kg-1), asiaticoside ointment group(5 mg·kg-1) and compound heparin sodium allantoin gel group(20 mg·kg-1), 7 rabbits per group. Except control group, the different drug about 0.5 mL had been applied the hypertrophic scar of rabbit ears once a day. After 42 days, the tissues of hypertrophic scar were obtained. Hematoxylin-eosin（HE）staining was used to observe the pathological changes of rabbit ear scar tissue and determine the scar hyperplasia index. The expression of TGF-β1 and Smad3 in scar tissue of rabbit ears were detected by immunohistochemistry, Western blot and reverse transcription PCR（RT-PCR）.ResultCompared with control group, the pathological changes of the ear scars in the model group showed obvious hyperplasia and higher hyperplasia index (P<0.01). Meanwhile, the expressions of TGF-β1 and Smad3 in scar tissue of rabbit ears were significantly increased (P<0.01). Compared with model group, the pathological structures of the ear scar tissue were significantly improved and the hyperplasia index of ear scar tissue was clearly reduced in medium and high-dose groups of ethanol extracts from Sophorae Tonkinensis Radix et Rhizoma(P<0.05，P<0.01). The protein and mRNA expression of TGF-β1 and Smad3 in scar tissue were also decreased in different group of ethanol extracts from Sophorae Tonkinensis Radix et Rhizoma compared with the model group (P<0.05，P<0.01).ConclusionsEthanol extracts from Sophorae Tonkinensis Radix et Rhizoma may play a curative role in inhibiting hypertrophic scars by reducing the expression of TGF-β1 and Smad3 in scar tissue and inhibiting the TGF-β1/Smads signal transduction pathway. These provides the experimental basis for the clinical application of Sophorae Tonkinensis Radix et Rhizoma in the treatment of hypertrophic scars.
Keywords：Sophorae Tonkinensis Radix et Rhizoma;TGF-β1/Smad signaling pathway;ethanol extracts;hypertrophic scar rabbit
Abstract：ObjectiveTo observe the efficacy of modified Yuquanwan combined with Taohong Siwutang on major cardiovascular risk factors of type 2 diabetes and its effect on inflammatory factors and endothelial function.MethodOne hundred and sixty patients were randomly divided into control group (80 cases) and observation group (80 cases) by random number table. The control group was given Tianqi Jiangtang capsule 5 tablets/time,3 times/day. Bothgroups were taken orally. aspirin enteric-coated tablets, 0.1 g/time,1 time/day, insulin for injection or oral antidiabetic, simvastatin tablets, 20 mg/time, 1 time/day, and irbesartan tablets, 150 mg/time, 1 time/day.patients in observation group were added with modified Yuquanwan combined with Taohong Siwutang for 24 weeks, 1 dose/day, and a 48-week follow-up visit were paid. Blood glucose, lipid, blood pressure, fasting plasma glucose (FPG), blood sugar 2 h after meal (2 h PG), glycosylated hemoglobin (HbA1c), systolic (SBP) and diastolic pressure (DBP) were detected for every 8 weeks, and at the 8th, 16th, and 24th week after treatment, up-to-standard HbA1c, LDL-C, SBP, DBP and all of those indexes were recorded, and levels of blood glucose, lipids and blood pressure were compared at different time points. During the treatment and within the 72-week follow-up, cardiovascular events, stroke events, peripheral vascular events and microvascular complications were recorded. And levels of tumor necrosis factor-α (TNF-α), homocysteine (Hcy), interleukin-6 (IL-6), hypersensitive C-reactive protein (hs-CRP), endothelin (ET-1) and nitric oxide (NO) were detected, and at the 8th, 16th and 24th week after treatment, body mass index (BMI) was recorded.ResultAt the 24th week after treatment, the compliance rate of HbA1c in observation group was 81.16%（56/69）, which was higher than 64.71%（44/68） in control group (χ2=4.701, P<0.05), and the compliance rate of SBP was 94.2%（65/69）, which was higher than 82.36%（56/68） in control group (χ2=4.662, P<0.05). At the 16th week and 24th week after treatment, the compliance rate of LDL-C were 79.71%（55/69） and 88.41%（61/69）, which were higher than 63.24%（43/68） and 70.59%（48/68） in control group (χ2=4.5642, χ2=5.108, P<0.05). At the 16th week, the comprehensive compliance rate (blood glucose, blood pressure, blood lipid) in observation group was 59.42%（41/49）, which was higher than 41.18% （28/68） in control group (χ2=4.559, P<0.05). At the 24th week, the comprehensive compliance rate in observation group was 69.57% （48/69）, which was higher than 51.47% （36/68） in control group (χ2=4.695, P<0.05). At the 16th week, the compliance rate of BMI was 60.87% （42/69）, which was higher than 39.71% （27/68）in control group (χ2=6.136, P<0.05). At the 24th week, the compliance rate of BMI was 72.46% （50/69）, which was higher than 52.94% （36/68） in control group (χ2=5.585, P<0.05). At the 16th week after treatment, levels of 2 h PG and HbA1cin observation group were lower than those in control group (P<0.05). At the 24th week after treatment, levels of FPG, 2 hPG, HbA1c, SBP and DBP were lower than those in control group (P<0.05). Levels of TNF-α, Hcy, IL-6, hs-CRP and ET-1 were lower than those in control group (P<0.01), while level of NO was higher than that in control group (P<0.01). During 72 weeks of observation period, the rate of adverse vascular events in observation group was 13.04%(9/69), which was lower than 30.88%(21/68) in control group (χ2=5.957, P<0.05).ConclusionIn addition to the conventional western medicine therapy, modified Yuquanwan combined with Taohong Siwutang can further control the main cardiovascular risk factors of patients with T2DM, improve the endothelial function of T2DM patients, inhibit the expression of pro-inflammatory factors, and reduce the incidence of adverse vascular events.
Keywords：type 2 diabetes;cardiovascular risk factors;vascular events;Yuquanwan;Taohong Siwutang;inflammatory factors;vascular endothelial function
Abstract：ObjectiveTo observe clinical effect of addition and subtraction therapy of Si Junzitang combined with Simotang to outlet obstructive constipation (OOC) after stapled trans-anal rectal resection (STARR).MethodOne hundred and twenty-four patients were randomly divided into control group (62 cases) and observation group (62 cases) by random number table. Patients in control group got Qirong Ruichang oral liquid, 20 mL/time, 3 times/day. After operation, patients in observation group got addition and subtraction therapy of Si Junzitang combined with Simotang, 1 dose/day. And courses of treatment in two groups were 4 weeks, and 8 weeks' follow-up was recorded. Before the operation and at the second and fourth week after treatment, and the eighth week of follow-up, scores of main symptoms of constipation and Longo ODS were graded. Before the operation and at the fourth week after treatment, levels of superoxide dismutase (SOD), malondialdehyde (MDA), constipation patients quality of life self-assessment scale (PAC-QOL), anorectal pressure, anal resting pressure (ARP), maximum anal systolic pressure (MSP), rectal defecation pressure (RSP), FSV, CRS and MTV were recorded. And incidence, recurrence, normal defecation, satisfaction at the fourth week after the operation and safety were evaluated.ResultThe clinical rate in observation group was better than that in control group (Z=2.096, P<0.05). At the second, fourth after treatment and eigh weeks' for follow-up, score of main symptoms of constipation and Longo ODS were both lower than those in control group (P<0.01). Levels of ARP, FSV, FSV, CRS and MDA were lower than those in control group (P<0.01), levels of MSP, RSP and SOD were higher than those in control group (P<0.01). Incidence and recurrence rate in observation group were 20.97% (13/62) and 4.84% (3/62) were all lower than 39.71% (24/62) and 16.13% (10/62) in control group (P<0.05). Normal defecation rate in observation group was 91.94% (57/62) higher than 80.65% (50/62) in control group, but there was no statistical significance in two groups. And total score of PAC-QOL and scores of each factor were all lower than those in control group (P<0.01). Then there was no adverse reaction related to the traditional Chinese medicine.ConclusionAddition and subtraction therapy of Si Junzitang combined with Simotang can reduce constipation symptoms and the degree of illness, improve the quality of life, reduce the incidence of postoperative complications and recurrence rate, and improve anorectal dynamic indicators and oxidative stress indicators, improve the clinical efficacy.
Abstract：ObjectiveTo observe the clinical efficacy of Shiwei Wendantang on post-ischemic stroke depression of heart and gallbladder Qi deficiency syndrome.MethodThe 80 patients with post-ischemic stroke depression of heart and gallbladder Qi deficiency syndrome were divided into two groups by random number table. 40 cases in control group received oral administration of antidepressant Paroxetine, 20 mg·d-1, and 40 cases in observation group received Shiwei Wendantang, 1 dose/day. The treatment course was 4 weeks in both groups. The clinical efficacy, hamilton depression scale (HAMD-17) score, serum C-reactive protein (CRP), homocysteine (Hcy) level, traditional Chinese medicine(TCM) syndrome score and quality of life score of two groups were observed and compared.ResultAfter treatment, the curative effect of observation group was better than that of the control group (Z=-2.104，P<0.05), and the total effective rate in observation group was significantly higher than that of control group（χ2 =5.00，P<0.05）. After treatment, the scores of each factor of HAMD-17 scale in observation group were significantly decreased (P<0.05), and the scores of factors of HAMD-17 scale in the control group were also significantly decreased (P<0.05) except despair and anxiety. The decrease of scores in the observation group was more obvious than that in the control group (P<0.05). After treatment, the values of serum CRP and Hcy in two groups were decreased significantly (P<0.01）,and the values of serum CRP and Hcy in observation group were also decreased significantly (P<0.01）.After treatment, the scores of TCM syndromes in observation group were decreased significantly (P<0.01）， while in control group, only the score of urination was significantly decreased（P<0.05）. The scores of quality of life in observation group were all significantly higher than those before treatment (P<0.01）. The scores of SF-36 Health Survey Form in the control group were significantly decreased except for energy and social function (P<0.05). The scores of physical function, physical intelligence, physical pain, social function, and mental health in observation group were significantly higher than those in the control group (P<0.05).ConclusionAs compared with the conventional western medicine treatment, the application of Shiwei Wendantang in the treatment of post-ischemic stroke depression of heart and gallbladder Qi deficiency syndrome, can further reduce the degree of depression of patients, improve the symptoms of depression, and improve the quality of life, with more significant clinical efficacy and comprehensive effect.
Keywords：Shiwei Wendantang;post-stroke depression;heart and gallbladder Qi deficiency;paroxetine
Abstract：ObjectiveMetabolomics was used to analyze the brain tissue samples of model mice with chronic unpredictable mild stress (CUMS) depression, in order to find out the differential metabolites related to depression and to explore the possible antidepressant mechanism of iridoid part of Valerianae Jatamansi Rhizoma et Radix (IEFV).MethodForty-two Kunming mice were randomly divided into 6 groups, including the normal group, the model group, the fluoxetine group (2.5 mg·kg-1) and the IEFV low, medium, and high dose groups (doses were 5.73, 11.47, 22.94 mg·kg-1, respectively). The behavioral and biochemical indicators of CUMS model mice were used for pharmacodynamic evaluation with IEFV and a positive drug (fluoxetine) as the intervention drugs. Then, the effect of IEFV on endogenous substances of the brain tissue in CUMS model mice were analyzed by nuclear magnetic resonance hydrogen spectrum (1H-NMR) metabolomics, and multivariate statistical analysis was used to identify the differential metabolites and to enrich the metabolic pathways involved in the differential metabolites.ResultAfter modeling, the immobility time of the model mice increased significantly, their sucrose preference rate and the excitatory neurotransmitters [5-hydroxytryptamine (5-HT) and norepinephrine (NE)] decreased significantly, indicating the success of modeling. The depression was relieved after IEFV administration, mainly manifested by the recovery of the immobility time, sucrose preference rate and the excitatory neurotransmitters (5-HT and NE). Principal component analysis (PCA) of endogenous metabolites in brain tissue showed that the model group could be significantly separated from the normal group, while the IEFV groups and fluoxetine group all showed a trend of deviating from the model group to the normal group, which was consistent with the behavioral results. The results of orthogonal partial least squares discriminant analysis (OPLS-DA) showed that there were 16 different metabolites between the model group and the normal group, including 12 water-soluble metabolites and 4 liposoluble metabolites. Seven potential metabolism pathways were obtained through MetPA analysis, including metabolism of phenylalanine, metabolism of phenylalanine, tyrosine and tryptophan, metabolism of taurine and hypotaurine acid, metabolism of alanine, aspartic acid and glutamic acid, biosynthesis of valine, leucine and isoleucine, metabolism of D-glutamine and D-glutamate and tricarboxylic acid cycle (TCA). IEFV-high dose group could significantly recall 11 differential metabolites.ConclusionIEFV may play an antidepressant role mainly by affecting energy metabolism, amino acid metabolism and neurotransmitter levels, which provides a reference for further study on the antidepressant mechanism of IEFV.
Keywords：Valerianae Jatamansi Rhizoma et Radix;iridoids;depression;chronic unpredictable mild stress (CUMS);brain tissue;nuclear magnetic resonance hydrogen spectrum (1H-NMR);metabolomics
Abstract：ObjectiveMetabolomics was used to analyze the dynamic changes of endogenous metabolites in serum and lung tissue of rats treated with water elution section of Siegesbeckiae Herba (SWES), and to explore the possible mechanism of lung injury and to search for the sensitive markers in serum and lung tissue.MethodSD rats were randomly divided into three groups, namely the normal group (normal saline), SWES high-dose group and SWES low-dose group (0.500, 0.125 g·kg-1·d-1). SWES was given for 4 weeks and stopped for 2 weeks. The weight and pathological examination were regarded as observation parameters. Serum and lung tissue samples were detected by nuclear magnetic resonance hydrogen spectrum (1H-NMR). The metabolites in rats were analyzed by partial least squares-discriminant analysis (PLS-DA) and orthogonal partial least squares-discriminant analysis (OPLS-DA).ResultLung inflammation was shown in SWES high-dose group and returned to normal after withdrawal for 2 weeks. The metabolic spectrum of SWES high-dose group was significantly different from the normal group. There were 11 metabolites were identified by 1H-NMR metabolomics, four differential metabolites were identified in serum [acetate, trimethylamine oxide (TMAO), glycine, myo-inositol] and nine differential metabolites were identified in lung tissue [lactate, acetate, phosphatidylcholine (PC), pyruvate, dimethylamine, glutamate, glycerophosphocholine (GPC), glycine, xanthine]. Lung injury was related to the disorder of pyruvate metabolism, glutamate metabolism and other pathways. Most of the metabolites in lung tissue had obviously levels of callback after drug withdrawal, which coincided with the pathological examination.ConclusionThe lung is one of the damaged organs caused by SWES, and the lung injury is reversible and may be related to energy metabolism and oxidative stress.
Abstract：ObjectiveTo establish a method for the determination of polysaccharide and monosaccharide composition of Tremella fuciformis, and to analyze the difference of polysaccharide content in T. fuciformis from different sources and cultivation methods, so as to provide reference for the quality determination.MethodHigh performance size exclusion chromatography coupled with multi-angle laser light scattering and refractive index detection (HPSEC-MALLS-RID) was employed to determine the content and relative molecular weight distribution of T. fuciformis polysaccharides. The monosaccharide types and proportions of T. fuciformis polysaccharides were analyzed by 1-phenyl-3-methyl-5-pyrazolone (PMP) precolumn derivative high performance liquid chromatography (HPLC).ResultThe weight-average relative molecular weight (Mw) and the content of polysaccharides in T. fuciformis cultivated by cut-log from different sources were distributed in 2.618×106-3.503×106 Da and 307.12-609.06 g·kg-1, respectively. These two parameters of polysaccharides in T. fuciformis with substitute cultivation from different sources were 2.723×106-3.886×106 Da and 366.38-647.37 g·kg-1, respectively. The T. fuciformis polysaccharides mainly consisted of mannose, glucuronic acid, glucose, galactose, xylose and fucose, their ratios in samples with cut-log and substitute cultivation were 4.4∶0.7∶1.0∶0.2∶1.4∶1.6 and 4.4∶0.8∶1.0∶0.1∶1.5∶1.5, respectively. The contents of the above six monosaccharides in 39 batches of T. fuciformis from different sources were mannose of 36.71-191.31 g·kg-1, glucose of 10.46-76.10 g·kg-1, galactose of 1.00-6.72 g·kg-1, xylose of 16.73-70.54 g·kg-1, glucuronic acid of 9.74-32.12 g·kg-1, fucose of 17.16-68.20 g·kg-1.ConclusionThe content of polysaccharides in T. fuciformis from different sources has a certain difference, the developed method can be used as a routine method for the quality evaluation of polysaccharides in T. fuciformis.
Abstract：ObjectiveTo screen out the active components of Platycodonis Radix-Armeniacae Semen Amarum, predict the targets and signaling pathways, construct the "multi-components, multi-targets and multi-pathways" interaction network and further investigate their molecular mechanism for the treatment of lung carcinoma based on network pharmacology.MethodActive components and corresponding targets of Platycodonis Radix-Armeniacae Semen Amarum were obtained through Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and literature consultation. Therapeutic targeted genes of Platycodonis Radix-Armeniacae Semen Amarum in the treatment of lung carcinoma were obtained from UniProt database and Genecards database. The "components-targets" network was constructed by using Cytoscape 3.6.0 software, and the protein-protein interactions network was constructed by String database and "Generate Style From Statistics" tool in Cytoscape software. Its molecular docking with active components of Platycodonis Radix-Armeniacae Semen Amarum was carried out by using Systems Dock Web Site network server. The Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment analysis on the therapeutic targets of Platycodonis Radix-Armeniacae Semen Amarum were performed with the Database for Annotation, Visualization Andintegrated Discovery (DAVID).ResultFourteen active components of Platycodonis Radix-Armeniacae Semen Amarum were screened out, including acacetin, cis-dihydroquercetin, spinasterol, licochalcone B, and luteolin, et al. One hundred and three therapeutic targets were screened out, including nitric oxide synthase 2 (NOS2), prostaglandin endoperoxide synthase 1 (PTGS1), androgen receptor (AR), prostaglandin endoperoxide synthase 2 (PTGS2), and dipeptidyl peptidase 4 (DPP4), et al. Identified signaling pathways mainly involved prostate cancer signaling pathway, small cell lung cancer signaling pathway, hepatitis B signaling pathway and T cell receptor signaling pathway.ConclusionThe possible molecular mechanism of Platycodonis Radix-Armeniacae Semen Amarum for the treatment of lung carcinoma was explored in this study based on network pharmacology, providing the direction for subsequent research.
Abstract：Gout is the second largest metabolic disease after diabetes, with a high incidence worldwide. Gout is a common and complex arthritic disease that, if left untreated, can damage joints and, in severe cases, lead to kidney stones and even life-threatening kidney failure. Although western medicine has also made significant achievements in the treatment of gout, it is often accompanied by gastrointestinal reactions, liver injury and other adverse reactions, and is prone to relapse after drug withdrawal, making the radical treatment of gout a difficult problem. Traditional Chinese medicine (TCM) used in gout have relatively long history, TCM has the characteristics of multiple ingredients targets, not only can produce related enzyme activity by inhibiting the uric acid(UA) and lowering uric acid generation, also can reduce uric acid by promoting uric acid excretion, except the uric acid reduction most applied in gout neighborhood of TCM can effectively reduce joint inflammation. TCM is mild, and the incidence of adverse reactions in the treatment of gout is significantly lower than that of western medicine. Some TCM can even play a role while protecting the kidney, so TCM is expected to solve the problem of treating gout. In recent years, a large number of studies have been conducted on the application of TCM in the gout neighborhood at home and abroad. By summarizing the studies on the application of TCM in the gout disease in the past 10 years, the mechanism of action and material basis have been summarized and analyzed, in the hope of providing references for the studies on the prevention and treatment of gout by TCM.
Keywords：gout;traditional Chinese medicine;mechanism;material basis