Network pharmacology-based study on compounds from Camptotheca acuminate inhibiting liver cancer cells

Camptotheca acuminate belongs to Nyssaceae family and has been used as traditional Chinese medicine for several years.A number of compounds with anti-tumor activities have been isolated from C.acuminate.However,the targets of some compounds have not been determined.In this study,network pharmacology methods were used to analyze 74 compounds from C.acuminate and 49 target proteins of liver cancer.The results showed that the target with the largest number of directed edges was Dihydroorotate Dehydrogenase(1d3g).The compound docked with 1d3g best was AL.23(19-O-methyangustoline).The results of this study provide further insight into the role of network pharmacology as an effective tool to illuminate the research on liver cancer.

Isolation and structural elucidation of the compounds from the fruits of Camptotheca acuminate

The fruit of Camptotheca acuminate is used as a traditional Chinese medicine.Eight compounds were isolated from the fruit of Camptotheca acuminate for the first time,including(+)-(S)-abscisic acid(1),blumenol A(2),(E)-3-hydroxymegastigm-7-en-9-one(3),(-)-(3R)-3,5-dimethyl-8-methoxy-3,4-dihydroisocoumarin(4),(-)-(3R)-8-methoxy-3-methyl-3,4-dihydro-1H-isochromen-1-one(5),neocnidilide(6),(3S)-butylphthalide(7)and senkyunolide I(8).Their structures were determined by comprehensive spectroscopic methods,including HRESIMS and 1D NMR.

Structure-based identification and pathway elucidation of flavonoids in Camptotheca acuminate

Flavonoid metabolism in Camptotheca acuminate remained an untapped area for years.A tandem MS approach was used and focused on the mining and characterizing of flavonoids in mature C.acuminate.Fifteen new flavonoids and forty-three known flavonoids,including fifteen flavone analogs,sixteen flavonol analogs,seven flavanone analogs,six chalcone analogs,four xanthone analogs,ten flavane analogs were mined and identified based on their MS/MS fragments.Fifty-three of them were firstly characterized in C.acuminate.Eight biosynthetic precursors for these flavonoids were also identified.We constructed a specific metabolic map for flavonoids according to their relative contents in the flowers,fruits,stems,and leaves of C.acuminate.Furthermore,the most probable genes involved in chalcone biosynthesis,flavonoid hydroxylation,methylation,and glycosylation were further mined and fished in the gene reservoir of C.acuminate according to their conserved domains and co-expression analysis.These findings enable us to acquire a better understanding of versatile flavonoid metabolism in C.acuminate.

Cloning and Sequence Analysis of Ma-14-3-3d Encoding a Homologue 14-3-3 Protein from Banana

[Objective] The aim of the study is to clone and analyze the gene encoding 14-3-3 protein from banana. [Method] Combined with PCR amplification, RACE (rapid amplification of cDNA ends) technique was employed to clone 14-3-3 gene from banana; then the amplified sequence was sequenced and homologically analyzed. [Result] A new cDNA homologous with 14-3-3 protein genes were obtained by RT-PCR and RACE ( rapid amplification of cDNA ends ) approaches. The full length of this cDNA was 866 bp encoding 197 amino acids. Alignment of deduced amino acid sequence with those from other plants revealed that the cDNA shared high homology with 14-3-3 protein genes from other plants, and was designated as Musa acuminata 14-3-3 gene (Ma-14-3-3d). Phylogenetic analysis reveals that Ma-14-3-3d has closer genetic relationship with those from monocotyledon species than those from other species. [Conclusion] Ma-14-3-3d belongs to the same lineage of 14-3-3 from monocotyledon.

香蕉柠檬酸合酶基因MaGCS的克隆及生物信息学分析

采用RACE-PCR的方法首次在香蕉中获得一个柠檬酸合酶基因的cDNA序列。经序列测定和Blastx比对分析表明,该cDNA全长1885bp,编码513个氨基酸残基,具有植物柠檬酸合酶基因的特征结构域,并与其他植物来源的乙醛酸循环体的柠檬酸合酶具有很高的序列相似性,将其命名为MaGCS(Musa glyoxysomal citrate synthase),并对其进行生物信息学分析,初步预测其理化性质及功能等。

Ri质粒诱导喜树毛状根培养基中喜树碱的研究

喜树(Camptotheca acuminate)可产生具有抗癌作用的喜树碱,有许多用细胞培养方法制备喜树碱的尝试,但都因极低的产率而无从深入。植物毛状根培养的应用将成为一种生物碱生产的替代方法。以发根农杆菌(Agrobacterium rhizogenes)A41.2556感染喜树诱导产生毛状根,建立喜树碱毛状根产生体系。甘露碱检测及PCR结果证实发根农杆菌Ri质粒的rolB基因已在喜树毛状根基因组中整合并得到表达。确定了用RP-HPLC测定发根培养基中喜树碱的的方法,比较了不同的吸附树脂对培养基中喜树碱的富集能力,从培养30d的喜树培养基中获得喜树碱干质量分数为0.14mg·g-1。

Condyloma acuminatum of the anal canal,treated with endoscopic submucosal dissection

Condyloma acuminatum(CA) is a common sexually transmitted disease caused by human papilloma virus infection. Not all individuals develop persistent, progressive disease, but careful follow up is required with moderate-to-severe dysplasia to prevent progression to malignancy. Standard therapies include surgical treatments(trans-anal resection and transanal endoscopic microsurgery) and immunotherapeutic and topical methods(topical imiquimod); however, local recurrence remains a considerable problem. Here, we report a case with superficial CA of the anal canal, treated with endoscopic submucosal dissection(ESD). A 28-year-old man presented with a chief complaint of hematochezia. Digital exam did not detect a tumor. Screening colonoscopy revealed 10-mm long, whitish condyles extending from the anal canal to the lower rectum. The lesion covered almost the whole circumference, and only a small amount of normal mucosa remained. Magnifying endoscopy with narrow band imaging showed brownish hairpin-shaped, coiled capillaries. Although histopathological diagnosis by biopsy revealed CA, accurate histological differentiation between CA, papilloma, and squamous cell carcinoma can be difficult with a small specimen. Therefore, weperformed diagnostic ESD, which provides a complete specimen for precise histopathological evaluation. The pathological diagnosis was CA, with moderate dysplasia(anal intraepithelial neoplasia 2). There was no recurrence at 16 mo after the initial ESD. Compared to surgical treatment, endoscopic diagnosis and resection could be performed simultaneously and the tumor margin observed clearly with a magnifying chromocolonoscopy, resulting in less recurrence. These findings suggest that endoscopic resection may be an alternative method for CA that prevents recurrence.

CO_2, H_2O exchange and stomatal regulation of regenerated Camptotheca acuminata plantlets during ex vitro acclimatization

For finding the changes in CO2, H20 exchange and their stomatal regulation during ex vitro acclimatization of regenerated Camptotheca acuminata plantlets, the net photosynthesis rate （Pn）, respiration rate （Ro）, light compensation point （Lc） and light saturation point （Ls）, transpiration rate （Tr）, stomatal conductance （gs） and water use efficiency（WUE） were measured during 37 days of ex vitro acclimatization. The results showed that Pn sharply increased until 29 days, then slightly decreased. A substantial decrease in Lc and a substantial increase of Ls in the former two weeks were observed, indicating the light regime enlargement for effective leaf photosynthesis. Tr and gs abruptly decreased during the first week then linearly increased until 29days ex vitro acclimatization, reflecting the strong regulation effect of stomata on water changes of ex vitro acclimating plantlets. Stomatal regulation effect on CO2 exchange was different from that on water exchange, i.e. P, was almost independent of gs during the first week, while P. was significantly correlated with gs thereafter （i.e. dual patterns）. Different from dual patterns of gs-Pn relation, the Tr monotonously linearly increased with gs. Furthermore, WUE was almost independent on gs during the first week, while a marked decreasing tendency with gs was found thereafter. At the beginning of the acclimatization, WUE was mainly determined by photosynthetic capacity, while transpiration becomes a main determinant factor for WUE from 7 to 37 days＇ acclimatization.

Characterization of Neopestalotiopsis clavispora , A New Etiological Agent of Leaf Spot Isolated from Banana

[Objectives]The study was to confirm the etiological agent of leaf spot on banana in Yunnan Province of China.[Methods]Fungus isolates were isolated from the diseased tissues,and cultured to observe the morphological characteristics of colony and spore.Furthermore,phylogenetic analyses and pathogenicity test were also conducted to confirm the pathogen.[Results]The fungus isolated from the diseased tissues was identified as Neopestalotiopsis clavispora.[Conclusions]N.clavispora is a new pathogen causing leaf spot of banana.This research provides the first description of N.clavispora as a causal agent on banana in China,and adds new insights related to the host range of N.clavispora.

Morphological and Molecular Identification of Curvularia geniculata Associated with Leaf Spot Disease of Banana in China

[Objectives]The paper was to identify the causal agent of leaf spot of banana in Hainnan,China.[Methods]Fungus isolates were isolated from the diseased tissues,and morphological characteristics of colony and spore were observed.Furthermore,phylogenetic analyses and pathogenicity test were conducted to confirm the pathogen.[Results]The fungus isolates from the diseased tissues was identified as Curvularia geniculata.[Conclusions]C.geniculata is a new pathogen causing leaf spot of banana in Hainan,China.This finding will help to broaden the distribution and host range of C.geniculata,indicating that it poses potential damage to banana in China.