Research progress on the material basis and pharmacological effects of Pien Tze Huang / 药学学报

The medicinal history of Pien Tze Huang is long, and it is the only "double top secret" variety of technology and formula at present. It has the effects of clearing heat and detoxifying, detumescence and pain, cooling blood and removing blood stasis. At present, researchers have analyzed and identified some compounds in Pien Tze Huang and its precious medicinal materials, such as Panax notoginseng, calculus bovis, snake gall and musk, and conducted activity screening, pharmacokinetics and pharmacological related studies on these chemical components. It was found that Pien Tze Huang had a significant effect on the treatment of acute and chronic hepatitis, ulcer, colon cancer, liver cancer and other diseases. The purpose of this paper is to systematically discuss the research achievements of researchers in recent years on the material basis, pharmacological effects and clinical application of Pien Tze Huang, with a view to providing ideas for the further research of Pien Tze Huang.

De novo transcriptome assembly and metabolomic analysis of three tissue types in Cinnamomum cassia / 中草药·英文版

OBJECTIVE@#The barks, leaves, and branches of Cinnamomum cassia have been historically used as a traditional Chinese medicine, spice, and food preservative, in which phenylpropanoids are responsible compounds. However phenylpropanoid biosynthesis pathways are not clear in C. cassia. We elucidated the pathways by descriptive analyses of differentially expressed genes related to phenylpropanoid biosynthesis as well as to identify various phenylpropanoid metabolites.@*METHODS@#Chemical analysis, metabolome sequencing, and transcriptome sequencing were performed to investigate the molecular mechanisms underlying the difference of active components content in the barks, branches and leaves of C. cassia.@*RESULTS@#Metabolomic analysis revealed that small amounts of flavonoids, coumarine, and cinnamaldehyde accumulated in both leaves and branches. Transcriptome analysis showed that genes associated with phenylpropanoid and flavonoid biosynthesis were downregulated in the leaves and branches relative to the barks. The observed differences in essential oil content among the three tissues may be attributable to the differential expression of genes involved in the phenylpropanoid and flavonoid metabolic pathways.@*CONCLUSION@#This study identified the key genes in the phenylpropanoid pathway controling the flavonoid, coumarine, and cinnamaldehyde contents in the barks, branches and leaves by comparing the transcriptome and metabolome. These findings may be valuable in assessing phenylpropanoid and flavonoid metabolites and identifying specific candidate genes that are related to the synthesis of phenylpropanoids and flavonoids in C. cassia.

Hemostatic efficacy of tranexamic acid and ε-aminocaproic acid in hip and knee arthroplasty:a Meta-analysis / 中国骨伤

OBJECTIVE@#To systematically evaluate the hemostatic efficacy of tranexamic acid and ε-aminocaproic acid in total hip arthroplasty (THA) and total knee arthroplasty (TKA).@*METHODS@#Randomized controlled trials (RCT) and retrospective case-control studies about tranexamic acid and ε-aminocaproic acid for the comparison of THA or TKA were searched electronically in PubMed, EMbase, Cochrane Library, China National Knowledge Infrastructure (CNKI), Wanfang, VIP from the time of building databases to July 2020. Two investigators carried out literature screening and data extraction according to the inclusion and exclusion criteria respectively. The methodological quality of the included randomized controlled studies was evaluated through the Cochrane Handbook, and the methodological quality of the included retrospective case-control studies was evaluated through the NOS scale. Blood loss, the incidence of thrombosis complications, per capita input of hemoglobin were Meta-analyzed by Review Manager 5.3 software.@*RESULTS@#A total of 6 articles were included, including 4 RCTs and 2 retrospective case-control studies. A total of 3 174 patients, including 1 353 in the tranexamic acid group and 1 821 in the ε-aminocaproic acid group. Meta-analysis results showed that there were no difference statistical significance in blood loss [MD=-88.60, 95%CI(-260.30, 83.10), P=0.31], blood transfusion rate [OR=1.48, 95%CI(0.96, 2.27), P=0.08], thrombotic complications [OR=0.80, 95%CI(0.07, 8.83), P=0.85], per capita hemoglobin input [MD=0.04, 95%CI(-0.02, 0.10), P=0.18] between tranexamic acid group and ε-aminocaproic acid group during THA. While in TKA, the blood loss of the tranexamic acid group was less than that of the ε-aminocaproic acid group [MD=-147.13, 95%CI(-216.52, -77.74), P<0.0001], the difference was statistically significant. The blood transfusion rate [OR=1.30, 95%CI(0.74, 2.28), P=0.37], thrombotic complications [OR=0.95, 95%CI(0.38, 2.36), P=0.92], per capita hemoglobin input [MD=-0.00, 95%CI(-0.05, 0.06), P=0.48], tourniquet time [MD=1.54, 95%CI(-2.07, 5.14), P=0.40] were similar between two groups, the difference was not statistically significant.@*CONCLUSION@#In THA, tranexamic acid and ε-aminocaproic acid have similar hemostatic effects, while in TKA, tranexamic acid can effectively reduce the patient's blood loss and has a better hemostatic effect. Tranexamic acid is recommended as one of the first choice hemostatic drugs for TKA.

Comparison of femoral tunnel and early curative effect analysis between arthroscopic three approach and traditional anteromedial portal for reconstruction of anterior cruciate ligament / 中国骨伤

OBJECTIVE@#To study the three-approach and traditional anterior medial technique to establish the femoral tunnel of position, length, and coronal angle and the early efficacy of anterior cruciate ligament reconstruction.@*METHODS@#Through retrospective research, from December 2018 to June 2019, a total of 36 patients diagnosed with simple anterior cruciate ligament tear and undergoing surgery were collected. All patients had a clear history of knee sprains and were divided into two groups. A group of 16 patients, including 11 males and 5 females, with an average age of (30.13±6.54) years and an injury time of 7 to 60 (30.19±15.78) days, three-approach technique was used to drill the femoral tunnel to reconstruct the anterior cruciate ligament. Another group of 20patients, including 15 males and 5 females, with an average age of (30.80±8.60) years, and an injury time of 7 to 60 (27.35±15.50) days, the traditional anterior medial approach was used to drill the femoral tunnel to reconstruct anterior cruciate ligament. CT 3D reconstruction technique was used to evaluate the femoral tunnel and the knee joint function was evaluated by Lysholm score of the knee joint.@*RESULTS@#All patients achieved primary healing after the surgical incision. No femoral tunnel fracture, vascular and nerve damage, difficulty in graft passage during the operation, and venous thrombosis occurred. All 36 patients were followed up on an outpatient basis, with a follow up period of 9 to 15 (12.00±2.83) months. Three-dimensional CT reconstruction was used to evaluate the femoral tunnel of the patients. The position of the femoral tunnel was described using the quartile method as the three-approach group:the lower (27.83±1.97) % of the femoral condyle and the posterior (25.57±3.20) %;the traditional approach group:the lower (28.38±3.21) % of the femoral condyle and the posterior (26.23±3.20) %. Bone tunnel length, three-approach group:(35.20±5.52) mm in total length, (23.20±2.07) mm in thick bone tunnel;traditional approach group:(34.60±4.26) mm in total length, (22.56±2.50) mm in thick bone tunnel. Coronal plane angle, three-approach group:(47.93±5.98) °;traditional approach group:(41.78±6.62) °. Knee joint Lysholm score, three-approach group:48.67±4.18 before surgery;97.00±2.48 at last follow up;traditional approach group:49.75±5.33 before surgery, 97.30±2.68 at last follow up, there were significant differences before and after surgery, no significant statistical difference between two groups.@*CONCLUSION@#The positions of the femoral tunnel drilled by the two methods were within the range of the anatomic stop of the anterior cruciate ligament, and there was no statistical difference. Compared with the traditional anterior medial approach, the coronal plane angle of the femoral tunnel drilled by the three-approach approach is relatively large, and there were no statistical differences in the length of the tunnel, the early postoperative effect of the two surgical methods, and the operation time. But the three approach has a wider and clearer vision. In addition, the knee flexion angle required for drilling the femoral tunnel during surgery is significantly smaller than that of traditional approach technology, which reduces the difficulty of surgery.

Cloning and functional verification of carboxyl CoA ligases(AeCCLs) in Arnebia euchroma / 中国中药杂志

Carboxyl CoA ligases(CCLs) is an important branch of adenylate synthetase gene family, which mainly has two-step catalytic reactions. Firstly, in the presence of adenosine triphosphate, it can catalyze the pyrophosphorylation of carboxylateswith diffe-rent structures to form corresponding acyl adenosine monophosphate intermediates. Secondly, adenosine monophosphate was replaced by free electrons in the mercaptan group of enzyme A or other acyl receptors by nucleophilic attack to form thioesters. In this study, on the basis of the transcriptome database of Arnebia euchroma, two genes were selected, named AeCCL5(XP_019237476.1) and AeCCL7(XP_019237476.1). Bioinformatics analysis showed that their relative molecular weights were 60.569 kDa and 60.928 kDa, theoretical PI were 8.59 and 8.92, respectively. They both have transmembrane domains but without signal peptide. By multiple sequence alignment and phylogenetic tree analysis, we found that the similarity between AeCCLs and other plant homologous proteins was not high, and the substrate binding sites of AeCCLs were not highly conserved. The reasons might be that the sequence and structure need to adapt to the changes of new substrates in the process of evolution. In this study, the full-length of AeCCL5 and AecCCL7 were cloned into the expression vector pCDFDuet-1. The proteins of AeCCL5 and AeCCL7 with His-tag were expressed in Escherichia coli. The proteins of AeCCL5 and AeCCL7 were purified by nickel column. In vitro enzymatic reactions proved that both AeCCL5 and AeCCL7 can participate in the upstream phenylpropane pathway of shikonin biosynthesisby catalyzing 4-coumaric acid to produce 4-coumarin-CoA, and then to synthesis p-hydroxybenzoic acid, which is an important precursor of shikonin biosynthesis in A. euchroma.

Effects of drought stress on physiological and biochemical and chemical components of Cinnamomum cassia seedlings / 中国中药杂志

Six month old Cinnamomum cassia seedlings were used to simulate drought stress with polyethylene glycol(PEG 6000). The physiological indicators(osmotic substances, antioxidant enzymes, etc.) and chemical components of seedlings under different drought levels and the correlation between the two were studied. The results showed that the chlorophyll content and relative water content decreased gradually with the increase of PGE 6000(0, 5%, 10%, 15%) concentration and time(3, 5, 7 d), while the soluble protein content, soluble sugar content and catalase(CAT) activity increased, but the rising rate slowed down with the time. The activities of peroxidase(POD), superoxide dismutase(SOD), malondialdehyde(MDA) and proline content increased at first and then decreased. The content of coumarin, cinnamaldehyde, cinnamic acid and dimethoxycinnamaldehyde decreased, while the content of cinnamyl alcohol continued to increase.Under drought stress, the fluorescence signals of reactive oxygen species and no contents in roots of C. cassia seedlings were significantly stronger than those of the control.Further correlation analysis showed that coumarin content, di-methoxycinnamaldehyde content and osmoregulation substance content were significantly negatively correlated(P<0.05), cinnamic acid content was significantly negatively correlated with POD and SOD activities(P<0.01).It was found that C. cassia seedlings showed a certain degree of drought tolerance under short-term or mild drought stress, but if the drought exceeded a certain degree, the physiological metabolism of the seedlings would be unbalanced.

Extraction and separation, structure analysis and biological activity of polysaccharides from Atractylodis Rhizoma / 中国中药杂志

Atractylodis Rhizoma(AR) is a traditional Chinese medicinal material for food and medicine, with the functions of eli-minating dampness, strengthening the spleen, expelling wind evil and dispersing cold. AR contains a variety of compounds, including sesquiterpenoids, alkynes, triterpenoids, aromatic glycosides, polysaccharides and so on. At present, the researches on AR mainly focus on volatile components, with relatively fewer researches on non-volatile components. Polysaccharide from Atractylodis Rhizoma(ARP) is an important material basis among non-volatile components for the efficacy. Due to its many biological activities such as immunomodulatory activity, anti-tumors, anti-virus and anti-oxidation, ARP has certain research value and potential. The diversity of the polysaccharide structure is the basis for biological functions, but it also increases the difficulty of carbohydrate research. The research on the extraction, separation, purification, structure and activity of ARP is in the preliminary exploration stage, still with many shortcomings. Herein, recent advancements in the extraction, purification, structural characteristics and biological activities of ARP were summarized in this article to provide scientific reference for the in-depth systematic research of ARP and the development of AR resources.

Reference gene screening for Real-time quantitative PCR in Polygonum multiflorum / 中国中药杂志

To select suitable references gene of Polygonum multiflorum for gene expression analysis in different tissues, five candidate reference genes like Actin,GAPDH,SAND,PP2A,TIP41 were selected from the transcriptome data of P. multiflorum, then the specific primers were designed. The expression stability of the five reference genes in different tissues of P. multiflorum was analyzed by Real-time quantitative PCR through avilable analysis methods such as geNorm, NormFinder, BestKeeper, Delta CT and RefFinder, to ensure the reliability of the analysis results. The results showed that there were significant differences in the expression levels and stability of candidate genes in different tissues of P. multiflorum. Ct distribution analysis of the expression levels of candidate genes showed that the expression levels of Actin and GAPDH genes were relatively high in different tissues, while the expression levels of SAND, PP2A and TIP41 were lower. The stability of each candidate gene was analyzed by different methods. The results of geNorm analysis showed that the expression of PP2A and GAPDH was the most stable, the expression stability of SAND was the worst, the stability of PP2A was the highest in both NormFinder and Delta CT, the stability of SAND was the lowest, and the stability of Actin was the most stable in BestKeeper analysis. Through the comprehensive evaluation and analysis of the stability of candidate genes by RefFinder, it is concluded that the stability of PP2A gene is the highest, followed by GAPDH, Actin, TIP41, SAND, and SAND gene is the worst. Therefore, the PP2A gene is an ideal reference gene for the analysis of gene expression in different tissues of P. multiflorum.

Pre-Transfusion Testing and Transfusion Strategy in Patients with Multiple Myeloma after Daratumumab Treatment / 中国实验血液学杂志

OBJECTIVE@#To investigate the procedure of pre-transfusion testing and transfusion strategy of patients with multiple myeloma (MM) treated by daratumumab (DarA).@*METHODS@#The blood samples of MM patients before and after DarA treatment from the Fifth Affiliated Hospital of Sun Yat-sen University were collected, and the ABO/Rh blood group antigen identification and DAT test results were compared. The results of antibody screening and cross matching of the patients before and after inactivation of red blood cells with 0.2 mol/L dithiothreitol (DTT) were compared and analyzed.@*RESULTS@#ABO/Rh blood group antigen typing showed no affecting in patients after treated by DarA; the result of DAT test showed negative. Irregular antibody screening showed that all the three cells(Ⅰ, Ⅱ and Ⅲ) were positive(1+~2+) and the self-control was negative. By microcolumn agglutination method, the main side of the multi-bag of blood showed no matched, while the secondary side showed all identical. After treated by DTT solution, the cross matching results in reagent red blood cells and the red blood cells of blood donors were both consistent, and the irregular antibody screening was negative. The K(+)O type erythrocytes used in parallel control were transformed into K(-)O type erythrocytes after DTT treatment. However, there was no significant changes in E(+) O type erythrocytes before and after DTT treatment. There was no condensation on the primary and secondary side of the condensed amine method. The primary and secondary sides of blood matching by saline method showed negative.@*CONCLUSION@#After treated by DarA, cross matching results from microcolumn agglutination method can be interfered by the residual drug antibody in MM patients, while the interference was eliminated in the presence of 0.2 mol/L DTT solution. However, no disturbance was observed when using condensed amine method or saline method. Therefore, corresponding transfusion procedures should be selected according to the emergency degree of blood transfusion to ensure the safety and timeliness of blood transfusion.

Medicinal Textual Research of Tsaoko Fructus / 中国实验方剂学杂志

Tsaoko Fructus is a type of food with the homology of medicine and food，and has a long history of cultivation and application in China. With the deepening of the development and utilization of Tsaoko Fructus，its economic value has appeared obviously，and relevant industries have great potential，with an important role in poverty alleviation in mountainous areas. However，the plants source of Tsaoko Fructus are confused in the process of actual production and resource utilization，and there is a lack of systematic report about the origin of varieties and medicinal history. The paper focused on a textual research for its names，plant sources，changes of the genuine producing areas and traditional efficacy through consulting the herbal works and the analysis of modern literature. The results showed that the medicine of Tsaoko Fructus was first recorded in ancient literatures of the Song dynasty，and there were many aliases and synonyms of the herb， with a great difference between ancient and modern times. In modern records，the medicinal alias names such as Hongcaoguo，Tucaoguo，Caoguoren are similar with the Chinese name of Caoguo，but they are not the same kind of plant and shall be distinguished. In the Ming dynasty，variety and source of the herb was complicated，and it was often recorded as the same class of medicinal materials with Amomi Fructus Rotundus and Alpiniae Katsumadai Semen，until it was listed as a class of medicinal material and distinguished from confounding medicines in the Qing Dynasty. The prescriptions containing Tsaoko Fructus include Caoguoyin，Caoguosan，Caoguowan，Caoguoshushui. Tsaoko Fructus could be used as medicine with peel or only kernel according to different diseases，and mainly processed with water or wine. In addition，there are other processing methods，such as frying，salt-processing， ginger- processing，simmering flour. Ancient literatures have basically the same records on properties，efficacies and indications of Tsaoko Fructus，considering it was warm in nature with the effect in relieving vomiting，eliminating food，intercepting malaria，expelling phlegm. This study provided a theoretical basis for its origin，quality control and resource development and utilization.

Effects of Exogenous Substances on Growth of Polyporus umbellatus Mycelium and Its Polysaccharide Content / 中国实验方剂学杂志

Objective:To explore the effects of diverse exogenous substances at different concentrations on the growth of<italic> Polyporus umbellatus</italic> mycelium and polysaccharide content and screen out the optimal growth condition for <italic>P. umbellatus</italic> mycelium， so as to provide a reference for its large-scale artificial cultivation. Method:<italic>P. umbellatus</italic> mycelium was cultured in media containing different exogenous substances using the method for fungal culturing in plate. The growth rate of the mycelium was judged by the colony diameter and the polysaccharide content was determined by the phenol-sulfuric acid method. Result:The high-dose cyclic adenosine monophosphate， 6-benzyl aminopurine （6-BA）， gibberellic acid （GA）， 2，4-dichlorophenoxyacetic acid （2，4-D）， vitamin （V） B₁， VB₃， VB₆， VB₉， and VB₁₂ all promoted the growth of <italic>P. umbellatus</italic> mycelium and elevated polysaccharides content. By contrast， indole acetic acid （IAA）， VC， and VB₂ inhibited its growth， with the most obvious inhibition detected in the high-dose VC group. IAA and VB₂ both reduced the polysaccharide content， whereas the high-dose VC significantly increased the polysaccharide content. Cyclic adenosine monophosphate， 6-BA， GA， 2，4-D， VB₁， VB₃， VB₆， VB₉， and VB₁₂ at the concentrations of 2 mmol·L^-1， 6 mg·L^-1， 15 mg·L^-1， 2 mg·L^-1， 4 mg·L^-1， 2 mg·L^-1， 4 mg·L^-1， 6 mg·L^-1， and 10 mg·L^-1， respectively， contributed to the growth of <italic>P. umbellatus</italic> mycelium<italic> </italic>and polysaccharide accumulation. Conclusion:The growth of <italic>P. umbellatus </italic>mycelium and polysaccharide accumulation can be regulated by adding exogenous substances to the culture medium.

Molecular Identification of 22 Species of Lauraceae by DNA Barcoding / 中国实验方剂学杂志

Objective:DNA barcodes suitable for Lauraceae plants were screened，and 22 Lauraceae plants were identified and classified. Method:The DNA of 22 species of Lauraceae was extracted and amplified by polymerase chain reaction（PCR） with different DNA barcoding primers，followed by electrophoresis and sequencing. Codon Code Aligner was used to proofread，splice， and remove the forward and reverse primer sequences. The sequence was imported into DNAMAN for multiple sequence alignment，and the base mutation sites were analyzed. The Kimura 2-Parameter（K2P） distance of different plants was calculated by MEGA，and the variation degree was analyzed according to the genetic distance. The phylogenetic tree was constructed based on the adjacency method. Result:Three pairs of DNA barcoding primers were used to amplify and sequence the DNA of 22 species of Lauraceae，and 20 species were identified by comparing the specific base sites of gene sequences<bold>.</bold> Conclusion:Four <italic>Litsea</italic> plants could be identified by <italic>matK</italic>， three <italic>Phoebe</italic> plants except for<italic> Cinnamomum validinerve </italic>by<italic> rbcL</italic>， and 20 Lauraceae plants by the combination of<italic> matK</italic>， <italic>rbcL</italic>， and <italic>rpoB</italic>，which provided a theoretical basis for the identification and development of Lauraceae plants. Among them，<italic>matK</italic> was predominant in the identification of Lauraceae plants，and the results also showed that the combination of sequences for plant identification could achieve a better result in DNA barcoding. This study investigated the genetic relationship between Lauraceae plants at the molecular level，aiming at providing a basis for the investigation，cultivation，development，protection， and utilization of medicinal plant resources.

Herbal Textual Research on Polygonati Rhizoma and Ethnic Usage / 中国实验方剂学杂志

Polygonati Rhizoma was first recorded in Mingyi Bielu（《名医别录》） as both medicine and food due to its high edible and medicinal values.However，there is no systematic textual research on its use as medicine and food. Polygonati Rhizoma and Polygonati Odorati Rhizoma show similar morphology，so they are often confused.Clematis apiifolia and Polygonatum odoratum are often regarded as the original plants of Polygonati Odorati Rhizoma. By reviewing the related records in Chinese herbal books of the past dynasties，this paper aimed to clarify the relationship between Polygonati Rhizoma，C.apiifolia and P.odoratum，trace the historical evolution of Polygonati Rhizoma as medicine and food from the name，harvesting and processing，property，flavor and meridian tropism，efficacy and indications，edible and medicinal records，and contraindications，and summarize its application in ethnic medicine，in order to inherit and develop the ethnic medicine. The findings demonstrated that the name of Polygonati Rhizoma and its synonyms were derived from its pictogram，function，and place of origin，containing a wealth of myth implication and Taoism thought.Polygonati Rhizoma，C.apiifolia and P.odoratum are three species，different from the conclusions in previous textual research. The seedlings，flowers，seeds，and rhizomes of Polygonati Rhizoma all could be used for edible and medicinal purposes，with its edible records exceeding the medicinal records. There were 22 medical books from the ethnic minority groups such as The Annals of National Medicine in Yunnan，Zhongguo Minzu Yaozhi Yao，and Dictionary of Chinese Ethnic Medicine recording the knowledge and application of 12 species in Polygonatum by 26 ethnic minorities.Due to their different living environments，lifestyles，and cultural customs，ethnic minorities have formed their unique medical cultures.

Selection and Validation of Appropriate Reference Genes of Cinnamomum cassia and Cinnamomum cassia var. macrophyllum / 中国实验方剂学杂志

Objective:To screen out stable internal reference genes suitable for real-time quantitative polymerase chain reaction（Real-time PCR） analysis of different parts of<italic> Cinnamomum cassia</italic> and <italic>C. cassia</italic> var. <italic>macrophyllum</italic>，in order to provide stable internal reference genes for gene expression analysis of three different parts of and <italic>C. cassia</italic> var.<italic> macrophyllum</italic> branches and leaves. Method:With 6 different tissues and organs， such as bark，branches and leaves of two plants of <italic>C. cassia</italic> and <italic>C. cassia</italic> var. <italic>macrophyllum</italic> as experimental materials，Real-time PCR technology was used to detect the five internal reference genes， namely glyceraldehyde-3-phosphate dehydrogenase（GAPDH），actin，ubiquitin-ligase enzymes（UBE），histone and tubin（TUB）. The analysis of the expression of the data. Furthermore， three commonly used internal reference gene analysis software，namely geNorm，NormFinder and BestKeeper，was used to analyze and evaluate the stability of the candidate internal reference gene. Result:The internal five reference genes were expressed in the bark，branches and leaves of the two plants，but with differences in stability. Comprehensive analysis showed that the expression stability of candidate internal reference genes was in the order of GAPDH>actin>UBE>histone>TUB. The internal reference genes of the two plants were analyzed separately，and the optimal internal reference gene was still GAPDH，indicating that GAPDH was the most suitable internal reference gene. TUB and histone ranked low in the three software，and should be eliminated in the screening of reference genes. They were not suitable for gene expression analysis of <italic>C. cassia </italic>and <italic>C. cassia</italic> var. <italic>macrophyllum</italic>. Conclusion:The most suitable internal reference gene for different parts of cinnamon，branches，and leaves of <italic>C. cassia</italic> and <italic>C. cassia</italic> var. <italic>macrophyllum</italic> was GAPDH. In this study，a screening system for internal reference genes of Real-time PCR of <italic>C. cassia</italic> and <italic>C. cassia</italic> var. <italic>macrophyllum</italic> was established to provide theoretical basis for studying functional regulation and expression of genes during the accumulation of effective components in different parts.

Screening of plant growth-promoting rhizobacteria and its effect on seed germination of Polygonum multiflorum / 中国中药杂志

In this study, the rhizobacteria and actinomycetes of Polygonum multiflorum were screened for the strains with indole acetic acid(IAA)-producing capacity by Salkowski method, the siderophore-producing strains by Chrome Azurol S(CAS) assay, and the strains with inorganic phosphorus-solubilizing capacity by PKO inorganic phosphorus medium. The strains were identified by morphological identification, physiological and biochemical characteristics, and 16 S rDNA sequences. Furthermore, the effect of growth-promoting strains on the seed germination and development of P. multiflorum was tested. The results showed that among 196 strains, two strains F17 and F42 were found to be capable of producing IAA and siderophore and solubilizing inorganic phosphorus simulta-neously. For F17 and F42, the results are listed below: 38.65 and 33.64 mg·L~(-1) for IAA production, 0.85 and 0.49 for siderophore-producing capacities(A_s/A_r), and 1.35 and 1.70 for inorganic phosphorus-solubilizing capacities(D/d), respectively. Comprehensive analysis revealed that strains F17 and F42 were identified as Pseudochrobactrum asacharolyticum and Bacillus aryabhattai, respectively, and both could significantly promote the seed germination of P. multiflorum.

Abdominal acupoint thread embedding therapy based on "brain-intestinal connection" for mild-to-moderate Alzheimer's disease and its effects on serum levels of APP and Aβ / 中国针灸

OBJECTIVE@#To compare the clinical efficacy of abdominal acupoint thread embedding therapy based on "brain-intestinal connection" combined with donepezil hydrochloride tablets and oral donepezil hydrochloride tablets alone for mild-to-moderate Alzheimer's disease (AD) and observe its effects on amyloid precursor protein (APP) and β-amyloid protein@*METHODS@#Sixty patients with AD were randomly divided into an observation group (30 cases, 3 cases dropped off) and a control group (30 cases, 3 cases dropped off). The patients in the control group were treated with donepezil hydrochloride tablets (5 mg per day); based on the treatment in the control group, the patients in the observation group were treated with abdominal acupoint thread embedding therapy at Zhongwan (CV 12), Xiawan (CV 10), Huaroumen (ST 24), Wailing (ST 26), Daheng (SP 15), etc., once every 10 days. Both groups were treated for 2 months. The mini-mental state examination (MMSE), Alzheimer's disease assessment scale-cognitive subscale (ADAS-Cog), activity of daily living scale (ADL), neuropsychiatric inventory questionnaire (NPI) as well as the serum levels of APP and Aβ@*RESULTS@#After treatment, the MMSE scores in the two groups were higher than those before treatment (@*CONCLUSION@#The abdominal acupoint thread embedding therapy based on the theory of "brain-intestinal connection" combined with donepezil hydrochloride tablets can improve cognitive function, self-care ability of daily life and mental behavior, and reduce the serum levels of APP and Aβ

Fingerprint Establishment ，Content Determination and α-glucosidase Inhibitory Activity Study of Polysaccharide from Desmodium styracifolium / 中国药房

OBJECTIVE：To estab lish the fingerprint ，analyze the monosaccharide composition and content ，investigate the inhibitory effects of the polysaccharide from Desmodium styracifolium on α-glucosidase in vitro . METHODS ：Polysaccharide from D. styracifolium was prepared by water extraction and ethanol precipitation. After hydrolyzed by TFA and derived by PMP ，HPLC method was adopted to establish the fingerprint （using glucose peak as reference ），and analyze the constituent and content of monosaccharide. The content determination was performed on Phenomenex Luna C 18 column with mobile phase consisted of acetonitrile-0.05 mol/L potassium phosphate （pH adjusted to 6.8 with sodium hydroxide ）in gradient elution at the flow rate of 0.8 mL/min. The detection wavelength was set at 250 nm，and column temperature was set at 30 ℃. The sample size was 10 μL. Using acarbose as control ，PNPG assay was used to investigate the α-glucosidase inhibitory activity of polysaccharide from D. styracifolium. RESULTS ：There were 9 common peaks in HPLC fingerprints of 18 batches of samples ，and the similarity of 15 batches of samples was higher than 0.90. Totally 7 peaks were identified as mannose ，rhamnose，galacturonic acid ，glucose， galactose，xylose and arabinose. The contents of rhamnose ，galacturonic acid ，glucose，galactose and arabinose were 0.471-2.092， 1.379-8.919，2.560-35.679，1.194-6.905，0.566-4.158 mg/g，respectively. Based on rhamnose ，the molar ratios of the other four monosaccharides were 1.58-4.07，2.26-19.95，2.20-4.21 and 1.31-2.86，respectively. The inhibitory activity of polysaccharide from D. styracifolium on α-glucosidase increased with the increase of dose ，and the half inhibitory concentrations of it was 0.70 mg/mL， lower than 7.76 mg/mL of acarbose （positive control ）. CONCLUSIONS ：Glucose is the main component of D. styracifolium polysaccharide in different batches ，and the contents of monosaccharides are different. The polysaccharide from D. styracifolium have significant inhibitory activity on α-glucosidase，which is better than that of acarbose.

Effect of Polygonum multiflorum-Andrographis paniculata intercropping system on rhizosphere soil actinomycetes community structure and diversity of P. multiflorum / 中国中药杂志

To investigate the effect of Polygonum multiflorum-Andrographis paniculata intercropping system on rhizosphere soil actinomycetes of P. multiflorum, the community structure and diversity of soil actinomycetes were studied by using the original soil as the control group and the rhizosphere soil actinomycetes communities of P. multiflorum under monoculture and intercropping systems as the experimental group. In this study 655 221 effective sequences were obtained with an average length of 408 bp. OTU coverage and rarefaction curve showed that the sequencing could represent the real situation of soil actinomycetes. According to the results of alpha diversity analysis, the diversity soil actinomycetes varied as follows: original soil>intercropping soil>monoculture soil. The soil actinomycetes community structure and the relative abundance of dominant genera were significantly changed by both monoculture and intercropping, especially monoculture. OTU clustering and PCA analysis of soil samples showed that all the soil samples were divided into three distinct groups and the original soil was more similar to intercropping soil. In addition, intercropping increased the relative abundance of some beneficial actinomyces, such as Kitasatospora and Mycobacterium, which was beneficial to maintain soil health and reduce the occurrence of soil-borne diseases. The results show that, P. multiflorum-A. paniculata intercropping reduced the change of community structure and the decrease of diversity of soil actinomycetes caused by P. multiflorum monoculture, and made the actinomycete community in rhizosphere soil of P. multiflorum close to the original soil.

Establishment of RNA interfered hairy root system of two CYP450 genes in Arnebia euchroma and its influence / 中国中药杂志

In this study, based on the transcriptome database of suspension cells of Arnebia euchroma, we explored two candidate cytochrome P450 enzyme genes that might relate to the shikonin biosynthesis downstream pathway when CYP76B74 sequence was referenced. We constructed interference-type hairy roots of candidate genes and cultured them. We measured the fresh weight, dry weight, total naphthoquinone content, shikonin and its derivatives content and expression levels of key enzyme genes involved in shikonin biosynthesis pathway. The effects of candidate genes on the growth and shikonin production of A. euchroma hairy roots were discussed, and the possible regulatory mechanisms that candidate genes affected shikonin synthesis were discussed. Through local Blast and phylogenetic analysis, two candidate CYP450 genes(CYP76B75 and CYP76B100) with high homology to CYP76B74 in A. euchroma were screened, and corresponding interference hairy roots were constructed. Compared with the control(RNAi-control), the fresh weight of CYP76B75 interfered hairy root(RNAi-CYP76B75) and CYP76B100 interfered hairy root(RNAi-CYP76B100) were significantly reduced, while dry weight were not affected, so the dry rate increased significantly. Except for β-acetoxyisovalerylalkannin, which is high in three groups of hairy roots, the contents of shikonin, deoxyshikonin, acetylshikonin, β,β'-dimethacrylicalkannin, β-hydroxyisovalerylshikonin,β-hydroxyisovalerylshikonin, isobutyrylshikonin and total naphthoquinones showed a consistent pattern: RNAi-CYP76B75>RNAi-CYP76B100>RNAi-control. Among them, the synthesis of β-hydroxyisovalerylshikonin was most significantly promoted by interfering with the expression of CYP76B75. The content of β-hydroxyisovalerylshikonin in RNAi-CYP76B75 was 11.7 times that of RNAi-control. RESULTS:: of real-time qPCR analysis showed that compared to RNAi-control, the expression levels of AePGT gene in RNAi-CYP76B75 and RNAi-CYP76B100 were not changed significantly, and the expression levels of CYP76B74 and AeHMGR were up-regulated. In addition, the expression level of CYP76B100 in RNAi-CYP76B75 was down-regulated, whereas in RNAi-CYP76B100, the expression of CYP76B75 was significantly up-regulated. Therefore, this study confirmed that when the expression of CYP76B75 and CYP76B100 were interrupted, the growth of hairy roots were suppressed, but the synthesis of shikonin were promoted. They might increase the shikonin biosynthesis by up-regulating the expression of CYP76B74 in the hairy roots of A. euchroma.

Research progress on the application of lower limb alignment in discoid meniscus injury / 中国骨伤

Discoid meniscus injury is a kind of common sports injury. Its treatment methods include arthroscopic discoid meniscus plasty, discoid meniscus subtotal resection, discoid meniscus total resection and so on. Although the short-term clinical effect is good, the long-term clinical effect is not ideal. At present, different scholars have different views on the choice of surgical methods for discoid meniscus injury. In recent years, many scholars have shown that the choice of operation and the change of lower limb force line are related to the therapeutic effect of discoid meniscus injury. This paper mainly summarizes the current situation of the treatment of discoid meniscus injury and the changes of the force line of the lower limbs after operation, and expounds therole of the evaluation of the force line of the lower limbs in the treatment of discoid meniscus, so as to provide the basis for the clinical individualized treatment of discoid meniscus injury.