Allotetraploidization event of Coptis chinensis shared by all Ranunculales

Coptis chinensis Franch.,also named Chinese goldthread is a member of Ranunculaceae in the order Ranunculales and represents an important lineage of early eudicots with traditional medicinal value.In our study,by using syntenic analysis combined with phylogenomic analysis of C.chinensis and four other representative genomes from basal and core eudicots,we confirmed that the WGD event in C.chinensis was shared by Aquilegia coerulea and Papaver somniferum L.and quickly occurred after Ranunculales diverged from other eudicots,likely a Ranunculales common tetraploidization(RCT).The synonymous nucleotide substitutions at synonymous sites distribution of syntenic blocks across these genomes showed that the evolutionary rate of the P.somniferum genome is faster than that of the C.chinensis genome by approximately 13.7%,possibly due to Papaveraceaes having an additional special tetraploidization event(PST).After Ks correction,the RCT dated to 115—130 million years ago(MYA),which was close to the divergence of Ranunculaceaes and Papaveraceaes approximately115.45—130.51 MYA.Moreover,we identified homologous genes related to polyploidization and speciation and constructed multiple sequence alignments with different reference genomes.Notably,the event-related subgenomes in the basal genomes all showed genomic fractionation bias,suggesting a likely allopolyploid nature of the RCT,PST and T-Alpha and T-Beta events in Tetracentron sinense.In addition,we detected that the sixteen P450 subfamilies were markedly expanded in the genomes of Ranunculales,and most of them were related to the RCT and PST events.We constructed a new platform for Early Eudicot Comparative Genomic Research(http://www.cgrpoee.top/index.html)to store more information.In summary,our findings support the WGD of C.chinensis shared by Ranunculales,which is likely an allotetraploidization event.This present effort offered new insights into the evolution of key polyploidization events and the genes related to secondary metabolites during the diversification of early eudicots.

Network pharmacology and molecular dynamics study of the effect of the Astragalus-Coptis drug pair on diabetic kidney disease

BACKGROUND Diabetic kidney disease(DKD)is the primary cause of end-stage renal disease.The Astragalus-Coptis drug pair is frequently employed in the management of DKD.However,the precise molecular mechanism underlying its therapeutic effect remains elusive.AIM To investigate the synergistic effects of multiple active ingredients in the Astragalus-Coptis drug pair on DKD through multiple targets and pathways.METHODS The ingredients of the Astragalus-Coptis drug pair were collected and screened using the TCMSP database and the SwissADME platform.The targets were predicted using the SwissTargetPrediction database,while the DKD differential gene expression analysis was obtained from the Gene Expression Omnibus database.DKD targets were acquired from the GeneCards,Online Mendelian Inheritance in Man database,and DisGeNET databases,with common targets identified through the Venny platform.The protein-protein interaction network and the“disease-active ingredient-target”network of the common targets were constructed utilizing the STRING database and Cytoscape software,followed by the analysis of the interaction relationships and further screening of key targets and core active ingredients.Gene Ontology(GO)function and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichments were performed using the DAVID database.The tissue and organ distributions of key targets were evaluated.PyMOL and AutoDock software validate the molecular docking between the core ingredients and key targets.Finally,molecular dynamics(MD)simulations were conducted to simulate the optimal complex formed by interactions between core ingredients and key target proteins.RESULTS A total of 27 active ingredients and 512 potential targets of the Astragalus-Coptis drug pair were identified.There were 273 common targets between DKD and the Astragalus-Coptis drug pair.Through protein-protein interaction network topology analysis,we identified 9 core active ingredients and 10 key targets.GO and KEGG pathway enrichment analyses revealed that Astragalus-Coptis drug pair treatment for DKD involves various biological processes,including protein phosphorylation,negative regulation of apoptosis,inflammatory response,and endoplasmic reticulum unfolded protein response.These pathways are mainly associated with the advanced glycation end products(AGE)-receptor for AGE products signaling pathway in diabetic complications,as well as the Lipid and atherosclerosis.Molecular docking and MD simulations demonstrated high affinity and stability between the core active ingredients and key targets.Notably,the quercetin-AKT serine/threonine kinase 1(AKT1)and quercetin-tumor necrosis factor(TNF)protein complexes exhibited exceptional stability.CONCLUSION This study demonstrated that DKD treatment with the Astragalus-Coptis drug pair involves multiple ingredients,targets,and signaling pathways.We propose a novel approach for investigating the molecular mechanism underlying the therapeutic effects of the Astragalus-Coptis drug pair on DKD.Furthermore,we suggest that quercetin is the most potent active ingredient and specifically targets AKT1 and TNF,providing a theoretical foundation for further exploration of pharmacologically active ingredients and elucidating their molecular mechanisms in DKD treatment.

Effects of Different Alkaloids in Coptis chinensis on Inhibiting TGEV Proliferation

[Objectives]To study the effects of different alkaloids in Coptis chinensis on inhibiting the proliferation of Transmissible gastroenteritis virus(TGEV).[Methods]The components and content of the main alkaloids in the extract of C.chinensis were analyzed.The main alkaloids were selected as drugs to inhibit the proliferation of TGEV.The maximum non-toxic concentration of Columbamine,Jatrorrhizine,Epiberberine,Coptisine,Palmatine,and Berberine was screened.The protective rate of each drug on TGEV-infected ST cells was determined,and the transcriptional inhibitory effect of the drug on TGEV N gene was detected by fluorescent quantitative PCR.[Results]The extract of C.chinensis mainly contains 6 alkaloids:Columbamine,Jatrorrhizine,Epiberberine,Coptisine,Palmatine,and Berberine,accounting for 2.03%,8.88%,9.21%,15.07%,14.63%,and 50.18%,respectively.In the range of the safe concentration,Jatrorrhizine,Palmatine,and Coptisine had better protective effects on ST cells infected with TGEV;compared with the Columbamine group,the cell protection rate was significantly different(P<0.05);compared with the Berberine group,the difference was extremely significant(P<0.01).The Coptisine and Palmatine groups had significant inhibitory effects on the transcription of TGEV N gene,and the difference was extremely significant compared with the virus group(P<0.05).[Conclusions]Jatrorrhizine and Palmatine in C.chinensis are the main components to inhibit the proliferation of TGEV.

Optimization of Compound Coptis Ointment Extraction through the Integration of Pharmacodynamic Indices and Orthogonal Test

[Objectives]To determine the optimal reflux extraction process conditions for Compound Coptis Ointment.[Methods]The study employed the orthogonal experimental design method and drug-sensitive disc agar diffusion method to evaluate the extraction rate,berberine hydrochloride content,and bacteriostatic ring diameter of the extract as comprehensive indices,and optimized the extraction process conditions of Compound Coptis Ointment using ethanol reflux.[Results]Based on the results of comprehensive indices,the optimal reflux extraction conditions for Compound Coptis Ointment were determined to be soaking in 1.5 times the amount of 70%ethanol for 2 h,followed by two extractions with 6 times the amount of 70%ethanol for 1 h each time.Three verification tests were conducted under the optimal process conditions.The yield of the extract was 28.32%±0.53%,the content of berberine hydrochloride was 4.68%±0.45%,and the diameter of the bacteriostatic ring was(2.5±0.2)cm.[Conclusions]The extract had higher drug content and exhibited better antibacterial effects.The optimized extraction process is simple,stable,and reliable,and can be effectively used to optimize the extraction process of Compound Coptis Ointment.

Correlation between Yield of Coptis chinensis Franch. and Soil Nutrients under Different Forests

[Objective] This study was conducted to investigate the correlation be- tween yield of Coptis chinensis Franch. and soil nutrients under different forests. [Method] The soil nutrients and yields of C. chinensis in fallow field, Cryptomeria japonica forest, Magnolia officinalis forest and Phellodendron chinense forest planted with C. chinensis were investigated. [Result] The contents of soil nitrogen （total ni- trogen, alkali-hydrolyzale nitrogen）, organic matter, potassium and phosphorous under the planting mode of C. chinensis under C. japonica forest were the highest; and the contents of organic matter and total nitrogen in the soils of M. officinalis and P. chinense forests were close to each other at the medium abundant level, and the phosphorous contents were more abundant than those in fallow field. [Conclusion] This study could provide a theoretical basis for scientific fertilization.

The chromosome-level reference genome of Coptis chinensis provides insights into genomic evolution and berberine biosynthesis

Coptis chinensis Franch, a perennial herb, is mainly distributed in southeastern China. The rhizome of C. chinensis hasbeen used as a traditional medicine for more than 2000 years in China and many other Asian countries. Thepharmacological activities of C. chinensis have been validated by research. Here, we present a de novo high-qualitygenome of C. chinensis with a chromosome-level genome of ~958.20 Mb, a contig N50 of 1.58 Mb, and a scaffold N50of 4.53 Mb. We found that the relatively large genome size of C. chinensis was caused by the amplification of longterminal repeat (LTR) retrotransposons. In addition, a whole-genome duplication event in ancestral Ranunculales wasdiscovered. Comparative genomic analysis revealed that the tyrosine decarboxylase (TYDC) and (S)-norcoclaurinesynthase (NCS) genes were expanded and that the aspartate aminotransferase gene (ASP5) was positively selected inthe berberine metabolic pathway. Expression level and HPLC analyses showed that the berberine content was highestin the roots of C. chinensis in the third and fourth years. The chromosome-level reference genome of C. chinensisprovides important genomic data for molecular-assisted breeding and active ingredient biosynthesis.

Study of Total Alkaloids from Rhizoma Coptis Chinensis on Experimental Gastric Ulcers

Objective： To study the effects of total alkaloids （TA） extracted from Rhizoma Coptis Chinensis on experimental gastric ulcer models. Methods： Four kinds of experimental ulcer models were established respectively by water-immersion stress, intragastric ethanol, acetic acid erosion, and pylorus ligation. The anti-ulcer effects of TA were evaluated, and compared with that of berberine （Bet） and cimetidine （Oim）. Results： TA showed significant inhibitory effects on ulcerative formation induced by water-immersion stress, intragastric ethanol, and pylorus l igation in dose-dependent manner, and showed therapeutic effect on acetic acid erosion-inducing ulcer, in comparison with the control group. The anti-ulcer activity of Bet was less than TA containing equal content of Bet. TA significantly reduced the free acidity, total acidity and total acid output, but didn＇t affect the gastric juice volume, gastric pepsin activity, adherent mucus quantity of stomach wall and free mucus dissolving in gastric juice. The suppressive activities of TA on gastric acid secretion didn＇t occur when it was administered into dodecadactylon at a dose of 360 mg/kg wt. Moreover, when compared with Oim, the inhibitory effect of TA on gastric acid secretion isn＇t proportional to the inhibitory effects on the formation of the 4 kinds of experimental ulcers. Conclusion： TA is a potent candidate in therapeutic drugs for treating gastric ulcer. Its anti-ulcer effective components and mechanism is not only related to Bet and inhibition of gastric acid, but also to other ingredients of TA and mechanism so far unknown.

Qualitative and Semi-quantitative Analysis of Quaternary Alkaloids in Coptis-scute Herb Couple by Electrospray Mass Spectrometry

A practical solution of qualitatively analyzing quaternary alkaloids in coptis-scute herb couple by electrospray ionization mass spectrometry（ES1-MS） was developed. Without the complicated pretreatment of sample, thc active ingredients including berberine, palmatine, coptisine, jatrorrhizine, epiberberine, and columbamine were identified and some relative content changing rules of alkaloids in coptis-scute couple were summarized in this article. The overall profiles of the complex extracts were obtained. After adding an internal standard（rutaecarpine）, semi-quantitative analysis was performed and the result indicates that the actual content of alkaloids was decreased by increasing the amount of scute. Based on the data obtained by high-performance capillary electrophoresis（HPCE）, the feasibility of semi-quantitative analysis by ES1-MS was further proved.

Coptis,Pinellia,and Scutellaria as a promising new drug combination for treatment of Helicobacter pylori infection

BACKGROUND Helicobacter pylori(H.pylori)is the most important infectious agent and plays an important role in the progression of chronic gastritis and the development of gastric cancer.AIM To identify efficient therapeutic agents or strategies that can treat H.pylori infection.METHODS We performed literature analysis,experimental validation,and network pharmacology.First,traditional Chinese medicine(TCM)prescriptions for the treatment of H.pylori infection were obtained from the China National Knowledge Infrastructure,China Biology Medicine,China Science and Technology Journal Database,and WanFang databases.In addition,we conducted a relevant search by Reference Citation Analysis(RCA)(https://www.referencecitationanalysis.com).Next,we used TCM Inheritance Support System V2.5 to identify core drug combinations in the TCM prescriptions.Then,an H.pylori-associated chronic mouse model of gastritis was established.The antibacterial properties and antiinflammatory potential of the core drug combination were evaluated by the rapid urease test,modified Warthin-Starry silver staining,histopathological analysis,and enzyme linked immunosorbent assay.Finally,the active compounds,hub targets,and potential signaling pathways associated with the core drug combination were analyzed by network pharmacology.RESULTS The TCM treatment of H.pylori was mainly based on reinforcing the healthy Qi and eliminating pathogenic factors by simultaneously applying pungent dispersing,bitter descending,cold and warm drugs.The combination of Coptis,Pinellia,and Scutellaria(CPS)was identified as the core drug combination from 207 prescriptions and 168 herbs.This drug combination eradicated H.pylori,alleviated the gastric pathology induced by H.pylori infection,and reduced the expression levels of tumor necrosis factor-α(P=0.024)and interleukin-1β(P=0.001).Moreover,a total of 35 compounds and 2807 targets of CPS were identified using online databases.Nine key compounds(tenaxin I,neobaicalein,norwogonin,skullcapflavone II,baicalein,5,8,2'-trihydroxy-7-methoxyflavone,acacetin,panicolin,and wogonin)and nine hub target proteins(EGFR,PTGS2,STAT3,MAPK3,MAPK8,HSP90AA1,MAPK1,MMP9,and MTOR)were further explored.Seventy-seven signaling pathways were correlated with H.pylori-induced inflammation and carcinogenesis.CONCLUSION In summary,we showed that CPS is the core drug combination for treating H.pylori infection.Animal experiments demonstrated that CPS has bacteriostatic properties and can reduce the release of inflammatory cytokines in the gastric mucosa.Network pharmacology predictions further revealed that CPS showed complex chemical compositions with multi-target and multipathway regulatory mechanisms.Although the results derived from network pharmacology are not necessarily comprehensive,they still expand our understanding of CPS for treating H.pylori infection.

Determination of coptis chinensis’ quality by FT-NIR spectroscopy

As a kind of traditional Chinese medicinal materials coptis chinensis has a lot of medicinal value and closely related to human health, Widely used in clinical or disease prevention and treatment. So the quality of medicinal coptis chinensis rapid determination is very important. Calibration model for determination of Coptis chinensis’ quality was built by partial least squares and near infrared spectroscopy, and the method of spectral pretreatment was discussed. Berberine and moisture content were used as evaluation index of Coptis chinensis’ quality according to traditional Chinese medicine standard in this experiment. The first derivative and vector normalization were adopted for the spectral preprocessing. The performance of the final model was evaluated according to root mean squares error of cross-validation (RMSECV), root mean square error of prediction (RMSEP) and correlation coefficient (R). According to the model of calibration set, the R values were 0.9298 and 0.8898, RMSECV were 0.239% and 0.302%. The R values of prediction set were 0.9347 and 0.9146, RMSEP were 0.104% and 0.330%, and prediction results were accurate and reliable. This method could be applied in rapid analysis of Coptis chinensis. The method was suitable for production fields and research fields, and had a wide application prospect.

Structural and Histochemical Features of the Slow-Growing Perennial Coptis chinensis Franch.(Ranunculaceae)

Huanglian(Coptis chinensis Franch.)is a slow-growing perennial medicinal herb with considerable economic value.This study aimed to determine the structural characteristics and the levels of berberine deposits in the organs and tissues of Huanglian using light and epifluorescence microscopy.The adventitious roots are composed of primary and secondary structures with endodermis,exodermis,and phellem.The rhizome structures are composed of primary and secondary structures with cuticle and phellem.The leaves are composed of sclerenchymatous rings,isolateral mesophyll,and thin cuticles.We detected berberine in the xylem walls of the roots and rhizomes as well as in the sclerenchymatous rings of the petioles.We postulate that as the exodermis is developed,the deposition of berberine in the xylem closest to the root tips may affect water and nutrient absorption and transfer.Leaf blades had a thin cuticle and isolateral mesophyll,suggesting shade tolerance.These structural and histochemical features suggest that Huanglian is adapted to the slow growing nature of a shady environment.

Study of dual-directional regulatory effect of Banxia （Pinellia ternata） and Huanglian （Coptis chinensis） drug pair on gastrointestinal movement of mice

Objective： To evaluate the regulatory effects of Banxia （Pinellia ternata, P） and Huanglian （Coptis chinensis, C） drugpair, derived from Banxiaxiexin soup, on the gastrointestinal movement of mice with functional dyspepsia. Methods：Mice were treated with different proportions of the P and C drug pair （1：1, 3：1, and 4：1） for 10 days, and subsequentlyinjected with atropine （ATR） or neostigmine （NEO）. The effects of the different proportions of P and C were evaluatedbased on the alvine advance rate. In addition, we used the same modeling method used in the first experiment andadministered P： C at ratio of 3：1 and at different doses respectively （4.68 g/L, 2.34 g/L, and 1.17 g/L）, and tested levelsof the gastrointestinal hormones, gastrin （GAS）, vasoactive intestinal polypeptide （VIP）, and somatostatin （SS） in thesmall intestinal tissue using an enzyme-linked immunosorbent assay. Results： In the groups of NEO-induced mice, P：Cat ratios of 1：1, 3：1, and 4：1 significantly reduced the alvine advance rate compared with the NEO model group （P =0.003, P = 0.012 and P = 0.021, respectively）. In the groups of ATR-induced mice, only P：1 at ratio of 3：1 significantlyincreased the alvine advance rate compared with the ATR model group （P = 0.007）. After exposure to P： C at ratio of 3：1and at different dose, the GAS level was lower in the low-, medium-, and high-dose NEO groups than that in the NEOmodel group （P = 0.001, P = 0.004, and P = 0.003, respectively）. The VIP levels were higher in the medium-andhigh-dose NEO groups than that in the NEO model group （P = 0.004 and P = 0.002, respectively）. In addition, the SSlevel increased in the NEO medium-dose group compared with that in the NEO model group （P = 0.002）. The GAS levelwas higher in the ATR medium- and high-dose groups than in the ATR model group （P = 0.007 and P = 0.021,respectively）. The VIP level was lower in the ATR low-, medium-, and high-dose than that in the ATR model group （P =0.001, P = 0.001, and P = 0.001, respectively）. Furthermore, the SS level was lower in the ATR medium- and high-dosegroups than that in the ATR model group （P = 0.001 and P = 0.006）. Conclusion： The PC drug pair bidirectionallyadjusted the NEO- and ATR-induced functional dyspepsia in mice by modulating GAS, VIP, and SS levels in theintestine.

An Economical Method for Preparative Purification of Five Alkaloids from Coptis Chinensis Franch by High-Speed Counter-Current Chromatography Using Singled Prepared Solvent System by GC

Coptis chinensis Franch, a widely used Traditional Chinese Medicine, shows various kinds of bioactivity. The major active components of the herb are considered to be alkaloids. Thus, preparative separation of these alkaloids is critical important for further pharmacology and mechanism studies. In the paper, five alkaloids from C. chinensis were purified by HSCCC using the solvent system composed of chloro-form-metha- nol-water (2:1:1, v/v/v) single prepared. The content of each solvent in solvent system were determined by gas chromatography (GC), then according the ratios of solvents in each phase to prepare the mobile and stationary phase respectively. And a comparative study was carried out between together preparation and single preparation of the solvent system. The purities and recoveries of all the products were over 98.5% and 92%. However, 134 mL chloroform, 336 mL methanol and 452 mL water were saved when the two phase were singled by GC. Our research showed an economical method for separating alkaloids from C. chinensis by HSCCC using the solvent system single prepared by GC.

Modulation of gut microbiota by berberine and decocted Coptis chinensis Franch.in a high-fat diet-induced metabolic syndrome rat model

Objective:To investigate the effect of decocted Coptis chinensis Franch.(Huanglian Jianji,黄连煎剂,HLJJ) and berbenne on the gut microbiota of the rats with metabolic syndrome (MS) induced by high-fat diet (HFD).Methods:Sixty Sprague Dawley rats were randomly divided into normal diet (ND) group (n =15)and HFD group (n =45).After 8 weeks of feeding,the HFD group was further divided into three groups:a group provided with a combination of HLJJ and HFD for 4 weeks,a group provided with a combination of berberine and HFD for 4 weeks,and a group supplied only with HFD for 4 consecutive weeks.Epididymal fat weight,serum high-density lipoproteincholesterohC (HDL-C),lowdensity lipoproteincholesterol-C (LDL-C) and irisin levels were measured.In addition,the V3-V4 region of the cecum microbiota 16S rRNA gene amplicon was sequenced on the Illumina MiSeq platform.Results:MS was successfully induced in the rats with HFD for 8 weeks.After 12 weeks,serum irisin levels in the HFD + berberine group were significantly increased compared with those in the HFD group (P <.05).The high-quality tags were delineated into 1149 operational taxonomic units at a 97% similarity level.Furthermore,at the genus level,Akkermansia muciniphila (A.muciniphila),Bacteroides,and Ruminococcus were markedly increased,whereas Candidatus arthromitus,Prevotella,Phascolarctobacterium were in great decrease in the HLJJ group and the berberine group,compared with the HFD group.The abundance of A.muciniphila,Ruminococcus gnavus and Bacteroides was significantly higher in the berberine group,whereas,the abundance of Oscillospira was lower in the berberine group,compared with the HLJJ group.Conclusion:HLJJ and berberine changed the gut microbiota in rats with MS.The mechanism underlying the treatments of berberine and HLJJ on HFD-induced MS is partly different.

Optimization ratio between Coptis chinensis and cinnamo of Jiaotai pill plus benzodiazepines for insomnia in China:a network meta-analysis

OBJECTIVE To explore the efficacy of Jiaotai pill(JTW)and JTW modified prescriptions(JTW^(+))plus benzodiazepines(BDZ)for insomnia in China by systematic review and network meta-analysis,especially optimization ratio(_(m:n))between Coptis chinensis and Cinnamo.METHODS Randomized controlled trials(RCTs)regarding JTW and JTW^(+)+BDZ for insomina were comprehensively searched in CNKI,VIP,Wanfang,and PubMed,Cochrane Library,OVID,Embase databases from their inception to December 2017.The risk of bias was assessed by the Cochrane collaboration′s tool and Jadad scale.Systematic review and network meta-analysis were performed to evaluate the relative risk(RR)plus 95%confidence interval(95%CI)or prediction interval(95%PrI),and draw surface under the cumulative ranking curves(SUCRA)by Stata 14.0 and RevMan 5.0 software.RESULTS A total of 13 RCTs(1 RCT was omitted by sensitivity analysis)with 1040 patients were included.Overall heterogeneity was acceptable,except publication bias.Based on classical meta-analysis,JTW and JTW_(m:n)^(+)+BDZ,such as JTW_(2:1)^(+)+BDZ,JTW_(5:1)^(+)+BDZ,JTW_(6:1)^(+)+BDZ,JTW_(10:1)^(+)+BDZ,had more efficacy than BDZ.Based on network meta-analysis,JTW and JTW_(m:n)^(+)+BDZ,such as JTW_(2:1)^(+)+BDZ,JTW_(6:1)^(+)+BDZ,JTW_(10:1)^(+)+BDZ,had more efficacy than BDZ;meanwhile,SUCRA showed the best rank of JTW/JTW+(BDZ<JTW_(m:n)^(+)+BDZ<JTW)for insomina,as same as that of JTW_(m:n)+(BDZ<JTW_(5:1)^(+)+BDZ<JTW_(10:1)^(+)+BDZ<JTW_(2:1)^(+)+BDZ<JTW_(6:1)^(+)+BDZ).CONCLUSION Based on the existing clinical evidence,JTW has efficacy for insomnia,but the optimization ratio between Coptis chinensis and cinnamo of JTW_(m:n)^(+)+BDZ for insomnia needs more supports.

Analysis on Quality of Coptis chinensis Grown under Different Woods

[Objectives] To analyze the quality of Coptis chinensis grown under woods of Magnolia officinalis Rehd. et Wils.,Platycladus orientalis( Linn.) Franco.,and Cunninghamia lanceolata( Lamb.). [Methods] The content of five kinds of alkaloids in C. chinensis grown under three different woods was determined by high-performance liquid chromatography( HPLC). Detection conditions: 0. 03 mol/L methanol and potassium dihydrogen phosphate solution( volume ratio of 65∶ 35) as the mobile phase,flow rate of 1. 0 mL/min,detection wavelength of 350 nm,and column temperature of 35℃. [Results] In these three woods,the quality of C. chinensis grown under P. orientalis woods was the highest. [Conclusions] This study is expected to provide certain references for cultivation and planting of C. chinensis under woods.

Sustainable livelihoods in a Coptis-planting based rural community:a case study in Shizhu County,China

Coptis chinensis is a medicinal herb used in traditional Chinese medicine.This research features a case study in Shizhu County,China.Participatory Rural Appraisal(PRA) is used as the main tool for data collection.The study demonstrates that the current method of Coptis planting leads to forest destruction.Even with reforestation measures,it leads to the loss of biodiversity in the forest.It also shows that farmers cannot get satisfactory economic returns from Coptis planting,as the return is unstable and unpredictable.One alternative to the current Coptis planting mode is to incorporate it into the agroforestry system.Water shield planting also serves as a good alternative to Coptis planting itself.Improved methods of management,post-harvest processing,and marketing of Coptis are also proposed and analyzed.The intended contribution of this paper to the theoretical debate of sustainability shall be the provision of an example showing how agricultural production affects forest conservation.It also presents redressable measures to the negative consequences.Besides,one alternative livelihood pathway for the rural community in this case study is suggested.

Investigation of the differences between the “COLD” and “HOT” nature of Coptis chinensis Franch and its processed materials based on animal’s temperature tropism

The description and differentiation of the so-called "Cold" and "Hot" natures, the primary "Drug Naure" of Chinese medicine, is the focus of theoretical research. In this study, the divergency between the "Cold" and the "Hot" natures was investigated through examining the temperature tropism of mice affected by Coptis chinensis Franch and its processed materials by using a cold/hot plate differentiating technology. After exposure to C. chinensis Franch, the macroscopic behavioral index of the remaining rate (RR) on a warm pad (40℃ ) significantly increased (P<0.05), suggesting the enhancement of Hot tropism. The internal indexes of adenosine triphosphatase (ATPase) activity and oxygen consuming volume decreased significantly (P<0.05), suggesting the decapability of energy metabolism. This external behavior of Hot tropism might reflect the internal Cold nature of C. chinensis Franch. However, the processed materials of C. chinensis Franch exhibited a different Cold nature in temperature tropism compared with crude C. chinensis Franch (CC): the Cold nature of bile-processed C. chinensis Franch (BC) enhanced while the ginger-processed C. chinensis Franch (GC) changed inversely. The changing sequence was consistent with the theoretical prognostication. It is indicated that the external Cold & Hot natures of Chinese medicine may possibly reflect in an ethological way for the changes of animal’s temperature tropism which might be internally regulated by the body’s energy metabolism.

Establishment and reliability evaluation of the design space for HPLC analysis of six alkaloids in Coptis chinensis(Huanglian) using Bayesian approach

Coptis chinensis(Huanglian) is a commonly used traditional Chinese medicine(TCM) herb and alkaloids are the most important chemical constituents in it. In the present study, an isocratic reverse phase high performance liquid chromatography(RP-HPLC) method allowing the separation of six alkaloids in Huanglian was for the first time developed under the quality by design(Qb D) principles. First, five chromatographic parameters were identified to construct a Plackett-Burman experimental design. The critical resolution, analysis time, and peak width were responses modeled by multivariate linear regression. The results showed that the percentage of acetonitrile, concentration of sodium dodecyl sulfate, and concentration of potassium phosphate monobasic were statistically significant parameters(P < 0.05). Then, the Box-Behnken experimental design was applied to further evaluate the interactions between the three parameters on selected responses. Full quadratic models were built and used to establish the analytical design space. Moreover, the reliability of design space was estimated by the Bayesian posterior predictive distribution. The optimal separation was predicted at 40% acetonitrile, 1.7 g·m L-1of sodium dodecyl sulfate and 0.03 mol·m L-1 of potassium phosphate monobasic. Finally, the accuracy profile methodology was used to validate the established HPLC method. The results demonstrated that the Qb D concept could be efficiently used to develop a robust RP-HPLC analytical method for Huanglian.

Protective Effects and Mechanism of Total Coptis Alkaloids on Aβ_(25-35) Induced Learning and Memory Dysfunction in Rats

Objective： To observe the effect of total coptis alkaloids （TCA） on β -amyloid peptide （A β 25-35） induced learning and memory dysfunction in rats, and to explore its mechanism. Methods： Forty male Wistar rats were randomly divided into four groups： the control group, the model group, the TCA low dose （60 mg/kg） group and the TCA high dose （120 mg/kg） group, 10 in each. A β 25-35 （5 μl, 2 μg/μl） was injected into bilateral hippocampi of each rat to induce learning and memory dysfunction. TCA were administered through intragavage for consecutive 15 days. Morris Water Maze test was used to assess the impairment of learning and memory; concentration of malondialdehyde （MDA） in cerebral cortex was determined by thiobarbituric acid reactive substance to indicate the level of lipid peroxidation in brain tissues; activity of manganese-superoxide dismutase （Mn-SOD） in cerebral cortex was determined by xanthine-oxidase to indicate the activity of the enzyme; and NF-κB protein expression in cerebral cortex was measured by SP immunohistochemistry. Results： （1） Morris Water Maze test showed that, during the 4 consecutive days of acquisition trials, the rats in the model group took longer latency and searching distance than those in the control group （P〈0.01）, which could be shortened by high dose TCA （P〈0.05）; during the spatial probe trial on the fifth day, the rats in the model group took shorter searching time and distance on the previous flat area than those in the control group （P〈0.01）, which could be prolonged after TCA treatment （for low dose group, P〈0.05; for high dose group, P〈0.01）. （2） Analysis of cerebral cortical tissues showed that, compared with the control group, MDA level got significantly increased and Mn-SOD activity decreased in the model group （both P〈0.01）. After having been treated with TCA, the MDA level got significantly decreased （P〈0.05 and P〈0.01 respectively for low and high dose group）, while relative increase of Mn-SOD activity only appeared in high dose group （P〈0.05）. （3） Immunohistochemistry analysis showed the protein expression of NF- κB got significantly increased after modeling, while high dose TCA can significantly inhibit it. Conclusion： TCA could improve A β 25-35 induced dysfunction of learning and memory in rats, and its protective mechanism is associated with its actions in decreasing MDA level, increasing Mn-SOD activity and inhibiting the expression of NF-κB in cerebral cortex.