Dendrobium offi cinale flowers flavonoids enriched extract protects against acute ethanol-induced gastric ulcers via AMPK/PI3K signaling pathways

Gastric ulcer is a widespread disease caused by various etiologies.Dendrobium offi cinale flowers exert several health benefits owing to their rich flavonoid content.In this study,protective effects and possible action mechanisms of D.offi cinale flowers’flavonoid enriched extract(DOFF)were assessed against gastric ulcer.The result of sodium nitrite-aluminum nitrate colorimetry showed that 52.34%of the total extractive was flavonoid,and ultra-high performance liquid chromatography time of flight mass spectrometer(UPLC-QTOF/MS)revealed the presence of 28 components in DOFF of which 14 belonged to flavonoids.In addition,in vivo assay revealed DOFF potential in reducing the formation of ethanol-induced gastric mucosal lesions,with drop-in ulcer index from 64.33±8.76 to 32.00±4.47.Similar results were revealed in human gastric mucosal epithelia cells,with cells viability to increase from 27.2%to 61.6%post DOFF administration.To analyzed the protect effect of DOFF,we used Western blotting and immunofluorometric assay to revealed the expression levels of key proteins in cell pathways.The results showed that DOFF(320μg/mL)could increase the level of oxidation marker protein(HO-1),apoptosis regulatory protein(Bcl-2)and autophagy marker(LC3β)by 50.84%,43.85%,and 59.21%compared with ethanol-treated group respectively.Further analyzed of the mitochondrial activity and apoptosis pathway,we found that DOFF appeared to mitigate against ethanolinduced gastric mucosal injury via AMPK/mTOR/ULK1 and PI3K/AKT autophagy signaling pathways.

Effect of Dendrobium nobile powder combined with conventional therapy on mild to moderate fatty liver

BACKGROUND Nonalcoholic fatty liver disease(NAFLD)encompasses a variety of liver conditions impacting individuals who consume minimal or no alcohol.Recently,traditional Chinese medicine has been gradually used to treat mild to moderate fatty liver,among which Dendrobium nobile Lindl.powder has been affirmed by many doctors and patients to be effective.However,there is limited research on combining this treatment with standard therapies for mild to moderate NAFLD.AIM To survey the effect of combining Dendrobium nobile Lindl.powder with standard treatment on liver function and lipid metabolism disorder in patients with mild to moderate NAFLD.METHODS Eighty patients with mild to moderate NAFLD participated in this retrospective study,classified into two groups:The observation group(n=40)and the control group(n=40).In November 2020 and November 2022,the study was conducted at People’s Hospital of Chongqing Liang Jiang New Area.The control group received standard treatment,while the observation group received Dendrobium nobile Lindl.powder based on the control group.The study compared differences in traditional Chinese medicine clinical syndrome scores,liver fibrosis treatment,liver function indicators,lipid levels,and serum inflammatory factor levels before and after treatment,and we calculated the incidence of adverse reactions for both groups.RESULTS The total effective rate was 97.50%in the observation group and 72.5%in the control group.After 8 weeks of treatment,the main and secondary symptom scores remarkably decreased,especially in the observation group(P<0.05),and there was a significant reduction in the serum levels of hyaluronic acid(HA),laminin(LN),human rocollagen III(PC III),and collagen type IV(CIV).The levels of HA,LN,PC III,and CIV were significantly lower in the observation group(P<0.05).After 8 weeks,both groups indicated remarkable improvements in liver function and blood lipid levels,with the observation group having even lower levels(P<0.05).Serum levels of interleukin-1β,tumor necrosis factor-α,and interleukin-8 also dropped significantly.The observation group had a lower rate of adverse reactions(5.00%)compared to the control group(22.50%).CONCLUSION Adding Dendrobium nobile Lindl.powder to standard treatment has been found to remarkably improve symptoms and reduce inflammation in patients with mild to moderate fatty liver disease.It also enhances hepatic function and lipid profile,ameliorates liver fibrosis indices,and lowers the risk of side effects.Consequently,this therapeutic protocol shows promise for clinical implementation and dissemination.

Identification and Molecular Characterization of the Alkaloid Biosynthesis Gene Family in Dendrobium catenatum

As one of the main active components of Dendrobium catenatum, alkaloids have high medicinal value. The physicochemicalproperties, conserved domains and motifs, phylogenetic analysis, and cis-acting elements of the genefamily members in the alkaloid biosynthesis pathway of D. catenatum were analyzed by bioinformatics, and theexpression of the genes in different years and tissues was analyzed by qRT-PCR. There are 16 gene families,including 25 genes, in the D. catenatum alkaloid biosynthesis pathway. The analysis of conserved domains andmotifs showed that the types, quantities, and orders of domains and motifs were similar among members ofthe same family, but there were significant differences among families. Phylogenetic analysis indicated that thegene family members showed some evolutionary conservation. Cis-acting element analysis revealed that therewere a large number of light-responsive elements and MYB (v-myb avian myeloblastosis viral oncogene homolog)-related elements in these genes. qRT-PCR showed that expressions of gene family members involved in alkaloidsynthesis were different in different years and tissues of D. catenatum. This study provides a theoretical basisfor further exploration of the regulatory mechanisms of these genes in the alkaloid biosynthesis of D. catenatum.

Anti-oxidation Activity in Vitro of Polysaccharides of Dendrobium Huoshanense and Dendrobium moniliforme

[ Objective ] The aim was to study the effects of polysaccharides of Dendrobium Huoshanense and Dendrobium moniliforme on oxygen free radicals and lipid peroxidation in vitro. [ Method] The scavenging action on .OH of polysaccharides, the inhibition action on O2^- of polysaccharides, MDA production of lipid peroxidation were measured by Fenton reaction, self oxidation system of C6H3（ OH）3 and TBA method respectively. [ Result] The half scavenging concentration on ·OH of potysaccharides of Dendrobium Huoshanense and Dendrobium moniliforme was 6.79 and 6.75 mg/ml respectively. As to their half inhibition concentration on O2 , the data was 3, 04 and 3.44 mg/ml respectively. Two kinds of Dendrobium polysaccharides bothhad inhibitory actions on lipid peroxidation of liver homogenate induced by auto-oxidation and inducedoxidation, and they could alleviate oxidation damages of mice liver mitochondria induced by Vc-Fe2＋ system. [Conclusion] Two kinds of Dendrobium polysacchaddes both had remarkable anti-oxidation activity in vitro.

Comparison of Stomatal Characteristics between Dendrobium loddigesii and Dendrobium candidum

[Objective]This study was to reveal the physiological and ecological mechanism of growth and development of test-tube seedlings.[Methods]Leaf stomata of test-tube seedlings of D.loddigesii and D.candidum at different growth and development stages were observed under scanning electron microscope,and their fluorescence induction response curve was assayed by using modulate fluorometer.[Results] At each growth and development stage,D.loddigesii test-tube seedling has a higher leaf stomatal density over D.candidum,while a lower stomata opening rate.Along with the growth,the size of stomata of both D.loddigesii and D.candidum did not change obviously,but their stoma density increased by 83% and 17.6% respectively.Leaf stomata opening rate reached the highest at the age of 240-day-old.Under the conditions of three settled light intensities,stomatal opening degree of D.loddigesii was increased by the intensity of illumination,the maximum aperture occurred at the light intensity of 54 μmol/m2·s.Given the same culture condition,both the maximum electron transport rate（14 μmol/m2·s）and light saturation point（318 μmol/m2·s）of D.loddigesii test-tube seedling are lower than that of D.candidum（20 μmol/m2·s,483 μmol/m2·s）.Moreover,the potential photosynthetic capacity of D.loddigesii is relatively poor.[Conclusion]The photosynthetic rate varies among different species of Dendrobium,which could be attributed to their different stomatal characteristics.Usually,the light intensity of tissue culture chamber is set up as 27 μmol/（m2·s）,where both D.loddigesii and D.candidum do not acquire an optimal state for their leaf stomatal opening.Considering the characteristics to select suitable culture condition,the stomatal aperture and photosynthetic rate could be enhanced,which can promote the growth and development of test-tube seedlings.

春石斛品种Dendrobium Second Love‘Tokimeki’与金钗石斛多糖免疫活性的比较研究

为比较春石斛栽培品种Dendrobium Second Love‘Tokimeki’与金钗石斛的粗多糖、精多糖(除蛋白多糖)和多糖水洗组分(过层析柱分离的水洗组分)的免疫活性差异,分别对昆明小鼠灌胃上述6种不同纯度的多糖,观察其对小鼠脏体指数、迟发性变态反应、抗体积数和小鼠腹腔巨噬细胞吞噬百分比与吞噬指数等免疫学指标的影响。结果表明:2种石斛的免疫活性随多糖纯度的提高而增强,纯度最高的多糖水洗组分免疫活性最强,且各指标均显著高于对照。D.Second Love‘Tokimeki’和金钗石斛同等纯度的多糖在胸腺指数、脾脏指数、小鼠耳肿胀率及小鼠抗体积数上均无显著差异,而D.Second Love‘Tokimeki’的3种不同纯度多糖的吞噬指数均达到了与金钗石斛多糖水洗组分无显著差异的水平。综合来看,栽培品种D.Second Love‘Tokimeki’的多糖免疫活性与金钗石斛多糖相近,具有良好的药用开发价值。

铁皮石斛(Dendrobium catenatum)快速繁殖的研究进展——兼论其学名与中名的正误

铁皮石斛是中国长期使用的名贵药用植物。近20年来,该药用植物的研究和生产已取得了显著的进展。本文系根据文献信息和在中国兰科植物种质资源保护中心的多年实验,对铁皮石斛快速繁殖研究的进展做一简要总结和评价。鉴于该种植物的学名和中名在不少文献中存在使用混乱,本文将在命名上和分类上予以澄清。笔者认为铁皮石斛的学名应当用其最早的合法名称Dendrobium catenatum Lindl.,而中名则应当用其最为常用的"铁皮石斛"。

Antioxidant phenanthrenes and lignans from Dendrobium nobile

To study the antioxidant constituents from the stems of Dendrobium nobile, and to discuss theft structure-activity relationship. Compounds were isolated from a 60% ethanolic extract by various chromatographic techniques and were identified by spectral analysis. The antioxidant activities of compounds were evaluated by DPPH free radical scavenging assay. Five phenanthrenes and four lignans were obtained from the active fractions ofD. nobile. Their structures were identified as fimbriatone （1）, confusarin （2）, flavanthrinin （3）, 2,5-dihydroxy-4,9-dimethoxyphenanthrene （4）, 3,7-dihydroxy-2,4-dimethoxyphenanthrene （5）, syringaresinol （6）, pinoresinol （7）, medioresinol （8） and lirioresinol-A （9）, respectively. Compounds 2 and 6 exhibited more potent DPPH scavenging activities than vitamin C. All the above compounds were reported from this plant for the first time, and compounds 3, 4 and 9 were reported for the first time from the genus of Dendrobiurn. For all phenanthrenes and lignans, an electron-donating methoxyl group in the ortho position to the phenolic hydroxyl group exhibits enhanced antioxidant activities.

Inoculation Effects of Dendrobium officinale Mycorrhizal Fungi on Their Plantlets

[Objective] This study aimed to explore the inoculation effects of Dendrobium officinale mycorrhizal fungi on their plantlets. [Method] Endophytic strains Tj1, Tj2 and Tj3 were obtained by isolation and purification from mycorrhiza of wild Dendrobium officinale and inoculated on the root system of Dendrobium officinale for inoculation test. [Result] Under tissue-culture conditions, at early stage, Tj1 strain hadn＇t shown promotion effect on Dendrobium officinale, Tj2 strain had shown relatively strong promotion effects, and Tj3 strain had promoted the growth of roots; at late stage, Tj1 strain had shown relatively strong promotion effects, Tj2 strain had shown the best inoculation effects and the strongest promotion effects, while Tj3 strain had caused root and seedling rot problems of the plantlets; under outdoor conditions, after inoculation with Tj2 strain, the number of leaves and lateral buds were increased, the growth of lateral root and the increase of plant height were significant, the leaves of Dendrobium officinale plantlets were large and dark green and an obvious root enlargement phenomenon was observed. [Conclusion] The two inoculation methods both indicate that Tj2 strain has relatively strong promotion effects on the growth of Dendrobium officinale roots and shoots, the increase of plant number and plant height, and the germination of new shoots and roots, which proved the effective establishment of symbiotic relationship between Tj2 strain and Dendrobium officinale. Therefore, T2 strain has practical application values on the successful cultivation of Dendrobium officinale plantlets.

Effects of Hormone Factors on the in vitro Culture Flowering Induction of Dendrobium officinate Kimura et Migo

[Objective]The aim was to select the suitable hormone factors for flowering induction in vitro culture of Dendrobium officinate Kimura et Migo.[Method]The test-tube plantlets from the stems of Dendrobium officinate Kimura et Migo were used as the experimental materials and MS medium as the basic medium.Comparative tests have been done between single-factor hormone treatments（different concentrations of PP333 or TDZ） and multi-factor hormone treatments（different combinations of PP333,TDZ,6-BA and NAA） to research the effects of hormone factors on the flowering induction of the plantlets.[Result]Among the single-factor hormone treatments,the suitable concentration and the rate of flower buds formation of PP333 treatment were 0.2 mg/L and 8.5%,the that of TDZ treatment were 0.06 mg/L and 15.5%;the effects of multi-factor hormone treatments on the flowering induction were ordered as follow：（PP333 ＋ 6-BA ＋ NAA ＋ TDZ）〉 （PP333 ＋ 6-BA ＋ NAA）〉 （PP333 ＋ 6-BA） and（PP333 ＋ NAA） ;the most suitable treatment was PP333 0.3 mg/L ＋ 6-BA 0.5 mg/L ＋ NAA 0.5 mg/L ＋ TDZ 0.06 mg/L,the rate of flower bud formation and the rate of the blossomed flower were respectly reached to 80.4% and 90.3%.[Conclusion]PP333 and TDZ showed the important effect on the flowering induction in vitro culture of Dendrobium officinate Kimura et Migo.The effect of TDZ was better than that of PP333.It is much more conducive to the flower bud formation,when using appropriate concentration of TDZ combined with other hormones properly.

Determination of Heavy Metals in Dendrobium candidium Wall.ex Lindl.by ICP-MS

[Objective] This research aimed to establish an inductively coupled plasma mass spectrometry （ICP-MS） method for determination of five kinds of heavy metals （Pb, Cd, Cu, As and Hg） in Dendrobium candidium Wall. ex Lindl. [Method] The samples were digested in HNO3-H202 acids system by closed vessel microwave di- gestion. At the same time, internal standard was added to avoid the matrix effect. [Result] The five kinds of heavy meals all showed good linear relationships, and the correlation coefficients ranged from 0.998 2 to 0.999 9. The detection limits of the five kinds of heavy metals were in the range of 0.008-0.750 μg/L, while the stan- dard recovery rates were in the range of 90.4%-96.9%. The relative standard devi- ations ranged from 1.6% to 8.2%. The heavy metals in the Ginseng and Astragalus national standard samples were determined by the established ICP-MS method, and the results were in conformity with the standard values. [Conclusion] The established method has simple and convenient operation and accurate and reliable results, and it meets the requirements by determination of heavy metals in Dendrobium candididium Wall. ex Lindl.

Optimization of Culture Conditions for Chromium-enriched Protocorm of Dendrobium candidum

[Objective] To optimize the techniques for culturing Cr-enriched Dendrobi-um candidum protocorm. [Method] By adopting Plackett-Burman test, the effects of time, temperature, Cr-content in medium, NAA content, KT content and light intensi-ty on Cr-enriched protocorm were researched. Three factors of time, Cr-content in medium and light intensity had statistical y significant effects; then steepest ascent procedures were applied to define optimal response region of these three factors; fi-nal y the optimal factors were determined by RSM analysis. [Result] The results showed that the optimal conditions for Cr-enrichment were chromium concentration at 0.37 mg/L, culture time of 59 d and il umination intensity of 1 822.22 lx. The predicted value of Cr-content in protocorm was 5.08 mg/kg. [Conclusion] RSM can optimize the techniques for culturing Cr-enriched protocorm of Dendrobium can-didum, provide the optimum process parameters and lay the foundation for further research.

Analysis of Genetic Diversity of Dendrobium by RSAP Marker

[Objective] This study aimed to analyse the genetic diversity of and genet- ic relationship between 33 Dendrobium species by using RSAP （Restriction Site Amplification Polymorphism） marker technique. [Method] A PCR-reaction system was established, by which 33 pairs of primers screened from 45 pairs were used to con- duct the PCR amplification. [Results] The 33 pairs of primers produced 2 047 ampli- fied fragments, of which 2 044 were polymorphic, with a polymorphism rate of 99.8%, and each pair of primers produced 61.9 polymorphic fragments on average. The genetic similarity coefficients between species ranged from 0.081 to 0.442, sug- gesting that the experimental materials used in this study possess a wide genetic background. According to the result of RSAP, the 33 Dendrobium species were di- vided into 6 groups by the cluster analysis （UPGMA）. The result showed that the genetic relationship between D. acinaciforme and D. guangxiense was the closest, and that between D. sinense and either D. lituiflorum or D. hancockii was the fur- thest. [Conclusion] The result suggested that Dendrobium germplasms have a great genetic diversity. This study provides a molecular basis for the classification and breeding of Dendrobium plants, as well as the selection of hybridization parents in the future.

The Interplanting Technology of Pleurotus ostreatus and Dendrobium officinale

In summer, Cha No. 39 of 292 bags and 205 of 244 bags were inter- planted with Dendrobium officinale on cultivation bed, respectively, whose yields and biological efficiencies reached 102.46 kg and 103.98%, and 96.77 kg and 113.32%, accordingly. The seedlings of Dendrobium officinale interplanted with Pleurotus os- treatus grew by 2.78 cm during 10 weeks on cultivation beds, and the seedlings grew by 2.68 cm in the cultivation beds without interplanting arrangement. These demonstrated that the interplanting of Dendrobium officinale and Pleurotus ostreatus has none adverse effects on growth of Dendrobium officinale.

Effects of Six Kinds of Fungal Elicitors on Growth of Dendrobium hybrida cultivar ‘088' Tissue Culture Seedlings

To optimize the technique of rapid propagation of Dendrobium hybrida seedlings and to explore a hormone-free tissue culture method for D.hybrida,six kinds of mycorrhizal fungi which were isolated from the wild orchids were made into fungal elicitors.These fungal elicitors were added into the DE medium with concentrations of 40,60 and 80 ml/L,respectively.After a 90-d culturing,the effects of fungal elicitors on the growth of D.hybrida cultivar ‘088＇ tissue culture seedlings were studied.The results showed that treatment 13（T13） extremely significantly increased the fresh weight,but other treatment groups had no significant effects.In addition,T1,T5,T9,T11 and T13 extremely significantly influenced the contents of chlorophyll a and chlorophyll a ＋ b.However,T1 and T11 had extremely significantly effect on the content of chlorophyll b.Combining the effects on fresh weight and chlorophyll content,it could be concluded T13（40 ml/L of Y05） has promoting effects on the growth of D.hybrida tissue culture seedlings.

Evaluation of Cold Tolerance of Different Dendrobium Species in North China

T o investigate cold tolerance of different cultivars and species in Dendrobium Sw. in North China, 8 strains of Dendrobium officinal and other 5 Dendrobium species were used to compare their cold-hardiness index in this study. The content of electrolyte leakage was increasing in Dendrobium Sw. as the temperature decreasing, while the content of peroxidase(POD) activity and the soluble sugar showed a rising trend before a downward trend. Besides, external morphology was another evaluation standard of cold tolerance. Thus, there was a significant difference between Dendrobium officinal and other Dendrobium species in the cold tolerance, and their cold tolerance capacity was as follows: s1 > s100 >s13 > 1301 > s300 > Dendrobium nobile > s126 > s145 > s95 > Dendrobium lindleyi > Dendrobium chrysotoxum > Dendrobium thyrsiflorum > Dendrobium lohohense.

A New Fluorenone Derivative from Dendrobium Chrysotoxum

A new fluorenone derivative, named chrysotoxone(3) was isolated from the ethanolic extract of the stems of Dendrobium chrysotoxum (Orchidaceae), along with three known compounds. The structure of the new fluorenone was elucidated as 2,5,8trihydroxy1,4dimethoxy9fluorenone.

Microwave Technique for Polysaccharide Extraction from Dendrobium huoshanense

The effect of material granularity, extraction times, extraction time, mi-crowave power and solid-liquid ratio on the yield of polysaccharide of Dendrobium huoshanense in microwave-assisted extraction was discussed through single factor experiment. The key factors in polysaccharide extraction and the advantages over the traditional extraction method were studied based on orthogonal experiment. The results showed that the influences became unobvious with the promotion of the fac-tor’s level. The orthogonal experiment showed that the optimal factor in the mi-crowave-assisted extraction was that the sample was extracted for four times with an extraction time of 40 min under 550 W. Al these results would lay a founda-tion for exploring a new method for the extraction of polysaccharide from Dendrobi-um huoshanense.

大苞鞘石斛(Dendrobium wardianum)种子离体萌发培养

通过无菌播种,对影响大苞鞘石斛(Dendrobium wardianum)种子萌发的各因素进行比较研究。结果表明,将种胚播于B5+20 g/L绵白糖+20%椰乳+pH 5.4上最有利于种胚萌发,培养50 d萌发率达79%。基本培养基、激素、氮源形态及有机添加物对大苞鞘石斛种胚萌发影响明显,萌发时需要硝态氮的量较大,并且基本培养基的正确选择可以有效地避免褐变的发生。

香豆素类成分在球花石斛(Dendrobium thyrsiflorum Rchb.f.)根中的分布和相对含量

利用激光共聚焦显微技术(laserscanningconfocalmicroscopy,LSCM)观测了香豆素类成分(coumarins)在球花石斛(DendrobiumthyrsiflorumRchb.f.)根中的分布及相对含量,为合理利用药用石斛提供科学的理论依据。观察结果显示香豆素类成分主要分布于球花石斛根的细胞壁,尤以细胞的角隅处为多;自发荧光强度以外皮层的最高,根被的最低。