A Case Report on Hepatopancreatic Anomaly with its histomorphological study in Adult Cadaver

The pancreas is a soft, lobulated and mixed gland. As the liver and pancreas develop together from the hepatopancreatic bud it is common that congenital anomalies of liver and pancreas exist together. This study was done during routine practical dissection where a band of tissue was found extending from the head of the pancreas to the root of the Mesentry. On histological examination the tissue was confirmed to be pancreas. It is seen that the hepatic bud grows at the expense of the ventral pancreas. In the present study, we found an atrophied left lobe of liver and an abnormally rare and large uncinate process of pancreas.

METTL3-modified RNASEH1-AS1 regulates the malignant biological behaviors of colorectal cancer SW480 cells through the BUD13/ANXA2/Wnt/β-catenin axis / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探究甲基转移酶样蛋白3（METTL3）修饰的RNASEH1-AS1通过BUD13/膜联蛋白A2/ Wnt/β-连环蛋白（BUD13/ANXA2/Wnt/β-catenin）轴调控结直肠癌细胞SW480恶性生物学行为的分子机制。方法：选取2022年6月至2022年11月间在复旦大学附属中山医院厦门医院手术治疗的24例CRC患者，收集患者的CRC组织和对应的癌旁组织，用qPCR法检测其中METTL3、RNASEH1-AS1、BUD13、ANXA2、β-catenin、GSK-3β mRNA的表达，用Pearson法分析CRC组织中RNASEH1-AS1表达分别与METTL3和BUD13表达的相关性。常规培养结直肠癌细胞SW480，分为sh-NC组、sh-RNASEH1-AS1组、NC组、sh-METTL组、si-NC组、si-BUD13组、sh-RNASEH1-AS1+pc-NC组、sh-RNASEH1-AS1+pc-ANXA2组、sh-METTL+pc-NC组、sh-METTL+pc-ASEH1-AS1组，用Lipofectamine® 2000转染试剂将sh-NC、sh-RNASEH1-AS1、sh-NC、sh-METTL、si-NC、si-BUD13、sh-RNASEH1-AS1+pc-NC、sh-RNASEH1-AS1+pc-ANXA2、sh-METTL+pc-NC、sh-METTL+pc-ASEH1-AS1分别转染SW480细胞，用qPCR法检测后SW480细胞中METTL3、RNASEH1-AS1、BUD13、ANXA2的表达，细胞克隆形成实验检测转染后各组SW480细胞的增殖能力，FCM术检测转染后各组SW480细胞的凋亡情况，细胞划痕实验检测转染后各组SW480细胞的迁移能力，WB法检测转染后各组SW480细胞中ANXA2、β-catenin、p-β-catenin、GSK-3β、p-GSK-3β、c-Myc、cyclinD1蛋白表达，RNA免疫沉淀（RIP）法检测RNASEH1-AS1与BUD1、BUD1与ANXA2的靶向关系。结果：数据库CRC数据分析和中国人CRC组织和癌旁组织的qPCR法检测结果均显示，与癌旁组织相比CRC组织中METTL3、RNASEH1-AS1、BUD13、ANXA2、β-catenin、GSK-3β均呈明显高表达（均P<0.01），且RNASEH1-AS1表达与METTL3（r=0.698，P<0.01）、BUD13（r=0.784，P<0.01）的表达呈正相关。在结直肠癌各细胞中METTL3、RNASEH1-AS1、BUD13、ANXA2 mRNA均呈高表达（均P<0.05）；敲减RNASEH1-AS1或METTL3后SW480细胞中RNASEH1-AS1、BUD13、ANXA2表达显著降低（均P<0.05），而过表达RNASEH1-AS1后上述分子表达明显上调（均P<0.05）；敲减RNASEH1-AS1或METTL3可抑制SW480的增殖、迁移和p-β-catenin、p-GSK-3β、c-Myc、cyclinD1蛋白的表达，促进其凋亡（均P<0.05），而过表达RNASEH1-AS1则可促进SW480增殖、迁移和p-β-catenin、p-GSK-3β、c-Myc、cyclinD1蛋白的表达和抑制其凋亡（均P<0.05）；RNASEH1-AS1通过招募BUD13靶向促进ANXA2的表达（均P<0.05）；过表达ANXA2可部分逆转敲减RNASEH1-AS1对SW480细胞的影响（均P<0.05）。结论：METTL3修饰的RNASEH1-AS1通过BUD13/ANXA2/Wnt/β-catenin轴调控SW480细胞的恶性生物学行为。

Topophysis and Growth Regulators Effects on Buds and Zygotic Embryos Regeneration of African Bush Mango Tree Cultured In vitro

Background: African bush mango tree is an important fruit plant with high nutritional, medicinal, and commercial values. However, its seedling system remained a deep understanding. This study aimed to evaluate the effect of topophysis and growth regulators on the reactivity of different types of buds and zygotic embryos of wild mango. Methodology: Ripe fruits from two local varieties (Wossro and Sissro) were pulped. The nuts were extracted and dried for one week under greenhouse. Zygotic embryos were excised from nuts and disinfected with the bleach solution (NaClO 10% with 8% active chloride) for 10 min following by three successive rinsing using sterilized water. The second set of nuts was cultivated for under greenhouse in the polybags containing the sand. One month later, buds from different positions (apical, axillary and cotyledonary) were excised and disinfected with NaClO 10% for 10 min follow by the immersion in a mercuric chloride 0.1% added with two drops of Tween 20 especially for axillary and cotyledonary buds for 5 min while 0.01% of mercuric chloride was used for apical buds. The explants were cultured on 糓S and 絎PM media supplemented with BAP, KIN at different concentrations (0.2 mg/L and 3.5 mg/L respectively) and NAA (0.05 mg/L). Results: The best sprouting rate was obtained with the variety Wossro which showed a high bud break rate (26.47%) against (9.88%) for the variety Sissro. The topophysis significantly influenced the response of the buds in tissue culture. 24.48% of axillary buds were sprouted on � MS medium + 3.5 mg/L BAP+ 0.05mg/L NAA. The in vitro germination of embryos was significantly (p? 0.05) influenced by the genotype of the variety. 50.76% of buds were germinated for Wossro while 18.32% were germinated for Sissro. � MS + 0.2 mg/L KIN has significant influenced the plants growth and development. Implication: The findings will help to improve the regeneration rate and plantlets production of African bush mango tree.

Dormancy of Floral Buds of Rabbiteye Blueberry in a Mild Winter Climate

Abstract Blueberry is an important fruit crop, with many health benefits. Despite its importance, much remains to be studied concerning the dormancy dynamics in rabbiteye cultivars growing in a mild winter climate. In this research, the dormancy in blueberry, rabbiteye cultivars 'Bluegem', 'Climax', 'Delite', and 'Powderblue', was studied in a mild winter region. The single-node cuttings biological test and the evaluation of the hydric status were performed in dormant winter reproductive buds. These experiments were performed during fall and winter in one year (2016). Moreover, chilling hours under or equal to 7.2 ºC were measured, and chill units were calculated according to Utah Model [1], Modified Utah Model [2], and Blueberry Model [3]. In conclusion, the four cultivars showed a similar pattern, revealing a dormant state in the initial sampling dates and a released dormancy in the final treatments, showing the decrease of dormancy in June and July. However, Delite was earlier than the other cultivars. Bluegem and Delite required 134.0 chilling hours, 127.0 chill units (Utah Model), 198.5 chill units (Modified Utah Model), and 971.5 chill units (Blueberry Model) for 50% of their green tip buds reach the opened bud stage. Climax required 44.0, -11.0, 56.5, and 440.5, respectively. And Powderblue required 44.0, 5.5, 77.0, and 725.0 respectively. This study can bring some insights into crop management and production of this important fruit crop, especially in a global climate-changing scenario, related to flowering and dormancy control, as well as helping to select suitable cultivars to a region, concerning chilling requirements.

Research Progress on Herbal Textual Research and Modern Quality Evaluation of Sojae Semen Germinatum / 中国实验方剂学杂志

Sojae Semen Germinatum was firstly recorded in Shennong Bencaojing， and it has a long history of edible and medicinal use. Most ancient medical practitioners described that Sojae Semen Germinatum was processed with black soya bean， while some others recorded that Sojae Semen Germinatum was processed with black soya bean and soybean or with soybean only. In modern times， black soya bean and soybean are both used. Before the Northern and Southern dynasties， the germination process of Sojae Semen Germinatum was mostly soil culture， and then changed into water culture later. The medicinal part of Sojae Semen Germinatum may also change from the initial aboveground part to the whole processed products including the soybean and the bud. The bud length was used to control the processing ending of Sojae Semen Germinatum， but there were different views of the bud length in ancient and modern times. Before the Tang dynasty， Sojae Semen Germinatum was mostly used directly. Since the Tang dynasty， various subsequent processed products of Sojae Semen Germinatum appeared. Most ancient medical practitioners confirmed that Sojae Semen Germinatum was sweet flavor， neutral in nature and non-toxic， and the mainstream believed that it belonged to the spleen， lung and stomach meridians. However， there were different opinions on its efficacy of relieving exterior syndrome by diaphoresis. In this paper， the evolution of Sojae Semen Germinatum was explored after consulting all kinds of ancient books， its historical name， processing history， quality evaluation and others were systematically summarized in order to clarify its historical development and lay a good foundation for the clinical use and further development of Sojae Semen Germinatum.

Early differential diagnosis between COVID-19 and mycoplasma pneumonia with chest CT scan / 浙江大学学报·医学版

OBJECTIVE@#To early differentiate between coronavirus disease 2019 (COVID-19) and adult mycoplasma pneumonia with chest CT scan.@*METHODS@#Twenty-six patients with COVID-19 and 21 patients with adult mycoplasma pneumonia confirmed with RT-PCR test were enrolled from Zibo First Hospital and Lanshan People's Hospital during December 1st 2019 and March 14th 2020. The early chest CT manifestations were analyzed and compared between the two groups.@*RESULTS@#The interstitial changes with ground glass density shadow (GGO) were similar in two groups during first chest CT examination (>0.05). There were more lung lobes involved on the first chest CT in COVID-19 patients, which were mostly distributed in the dorsal outer zone (23/26, 88.5%), and nearly half of them (12/26, 46.2%) were accompanied by crazy-paving sign; while the lesions in adult mycoplasma pneumonia patients were mostly distributed along the bronchi, and the bronchial wall was thickened (19/21, 90.5%), accompanied with tree buds / fog signs (19/21, 90.5%). The above CT signs were significantly different between the two kinds of pneumonia (all <0.01). COVID-19 had a longer course compared with mycoplasma pneumonia, the disease peaks of COVID-19 patients was on day (10.5±3.8), while the disease on CT was almost absorbed on day (7.9±2.2) in adult mycoplasma pneumonia. The length of hospital stay in COVID-19 patients was significantly longer than that of mycoplasma pneumonia patients [(19.5±4.3) d vs (7.9±2.2) d, <0.01].@*CONCLUSIONS@#The lesions of adult mycoplasma pneumonia are mostly distributed along the bronchi with tree buds/fog signs, while the lesions of COVID-19 are mainly distributed in the dorsal outer zone accompanied by crazy-paving sign, which can early distinguish two diseases.

Application of plant medicine containing spermine in pingyangmycin fermentation / 中草药

Objective: Screening a plant medicine containing spermidine to promote the fermentation of pingyangmycin (PYM). Methods: Based on the screening method of carbon and nitrogen sources in biological fermentation, the plant medicine with the best promotion effect on PYM was screened out by HPLC method in Lycium ruthenicum, Lycium barbarum, Lycium barbarum bud, Carthamus tinctorius, and Coicis Semen. And the best adding time, amount and frequency were also screened out. Results: The results showed that Lycium barbarum bud had the best effect on PYM fermentation, and the initial adding time was 24 h, the adding interval was 24 h, and the adding frequency was three times. The shaking bottle fermentation level was 21.3 μg/mL when the addition amount was 7 g/L/per batch, and the 25 L tank fermentation level was 37.5 μg/mL when the addition amount was 18 g/L/per batch, which increased by 23.8% and 118.0%, respectively. Compared with the addition of spermidine, the yield was increased by 45.3%. Conclusion: Most plant drugs containing spermidine can promote the fermentation of pingyangmycin, and the selection of Lycium barbarum bud as raw materials for pingyangmycin fermentation meets the production requirements in terms of cost, yield and environmental protection.

Callus Induction from Amomum villosum / 中国实验方剂学杂志

Objective: To set up a callus induction system for Amomum villosum by tissue culture. Method: The rhizome buds of A. villosum and stem segments,root tip segments of sterile A. villosum plantles were used as explants and cultured in MS media with different concentrations of 6-BA,NAA and 2,4-D (the pH of each medi is about 5.8). A callus induction system was established to explore the effect of different explants and different medium on callus induction for A. villosum. Result:The findings showed that the rhizome buds and sterile plantlet stems and root tip segments of three different explants can be successfully induced into calli. The most suitable medium for callus induction from rhizome buds and sterile plantlet stems was MS with 6-BA (1.5 mg·L-1),2,4-D (1.0 mg·L-1) and NAA (0.5 mg·L-1) with the highest induction rates of 15% and 60% respectively. MS medium combined with 6-BA (2.0 mg·L-1),2,4-D (1.0 mg·L-1) and NAA (1.0 mg·L-1) was the most suitable proposal for inducing the callus from sterile root tip segments with the highest induction rate of 76%. Conclusion:Under certain culture conditions,rhizome buds,stem or root tip segments of sterile plantlet can be effectively induced into callus. The callus induction system of A. villosum is preliminarily established, and root tip segments of sterile plantlet are the optimal explant.

Effect of irrigation at critical stages on the phenology of flowering and fruiting of the cactus Opuntia spp / Efeito da irrigação em estágios críticos sobre a fenologia de floração e frutificação do cacto Opuntia spp

Abstract This paper briefly reports some effects of irrigations at two critical periods on the phenology of three varieties of cactus pear cultivated in Agadir area: the spineless varieties 'Aissa' and 'Moussa' and the thorny one 'Achefri'. In the first year experiments (2010-2011) treatments of irrigation used were: (T1) 0 mm, (T2) 30 mm during flowering and 30 mm during fruit enlargement and (T3) 30 mm only during fruit enlargement. In the 2nd year experiments, irrigation treatments were: (T1) 0 mm, (T2) 60 mm during flowering and 60 mm during fruit enlargement and (T3) 60 mm only during fruit enlargement. Treatments of irrigation were applied between mid-April and mid-June in the 1st year experiments and in February and May in the 2nd year experiments. Results of the first year experiments showed that the emission of buds was higher in the thorny variety than in the spineless ones (more than 6 emitted buds/cladode vs less than 4.5 in the spineless varieties). In the second year, irrigation increased the emission of buds in the three varieties (more than 7 emitted buds/cladode for each T2 and T3 of all varieties vs not more than 5 for T1) and the duration of the flowering phase of these varieties. However, irrigation did not modify the proportions of fruits reaching commercial maturity during the early or the late period of maturation.

Resumo Este artigo relatou brevemente alguns efeitos das irrigações em dois períodos críticos na fenologia de três variedades de Opuntia cultivadas na área de Agadir: as variedades sem espinhos 'Aissa' e 'Moussa' e um espinhoso 'Achefri'. No primeiro ano (2010-2011) os seguintes tratamentos do experimento da irrigação foram usados: (T1) 0 milímetros, (T2) 30 milímetros durante a florescência e 30 milímetros durante a ampliação do fruto e (T3) 30 milímetros somente durante a ampliação do fruto. No segundo ano das experiências, os tratamentos da irrigação foram: (T1, 0 milímetros), (T2) 60 milímetros durante a florescência e 60 milímetros durante a ampliação do fruto e (T3) 60 milímetros somente durante a ampliação do fruto. Os tratamentos da irrigação foram aplicados entre a metade de abril e metade de junho nas experiências do primeira ano e em fevereiro e maio nas experiências do segundo ano. Os resultados das experiências do primeiro ano mostraram que a emissão de brotos foi maior na variedade espinhosa do que no rufia (mais de 6 emitida gomos/cladode vs inferior a 4,5 no rufia variedades). No segundo ano, a irrigação aumentou a emissão dos botões nas três variedades (mais de 7 botões emitidos/cladÓdio para cada T2 e T3 de todas as variedades não mais que 5 para o T1) e a duração da fase de florescência destas variedades. Contudo, a irrigação não alterou as proporções de frutos que alcançaram a maturidade comercial durante o período adiantado ou atrasado de maturação.

Ectopic Hypoplastic Gall Bladder- Case Report

Gallbladder, a derivative of foregut is a flask shaped structure present on the inferior surface of liver.it stores and concentrates bile. The diseases of this organ can be diagnosed by ultrasonography and operated through laparoscopy.But congenital anomalies are difficult to diagnose through scan and may lead to iatrogenic injury to biliary system.In order to avoid this a thorough knowledge of congenital variations is necessary. This is a case of rare congenital anomaly,in which a hypolpastic gallbladder was covered by liver tissue in the gall bladder fossa. This intrahepatic gallbladder is due to abnormal migration or incomplete canalization of cystic bud. Awareness of rare congenital anomalies will help the radiologist and surgeons to diagnose and plan their operative procedures which are either laparoscopic or open cholecystectomy .

Tree-in-Bud Pattern of Pulmonary Tuberculosis on Thin-Section CT: Pathological Implications

The “tree-in-bud-pattern” of images on thin-section lung CT is defined by centrilobular branching structures that resemble a budding tree. We investigated the pathological basis of the tree-in-bud lesion by reviewing the pathological specimens of bronchograms of normal lungs and contract radiographs of the post-mortem lungs manifesting active pulmonary tuberculosis. The tree portion corresponds to the intralobular inflammatory bronchiole, while the bud portion represents filling of inflammatory substances within alveolar ducts, which are larger than the corresponding bronchioles. Inflammatory bronchiole per se represents the “tree” (stem) and inflammatory alveolar ducts constitute the “buds” or clubbing. “Clusters of micronodules”, seen on 7-mm thick post-mortem radiographs with tuberculosis proved to be clusters of tree-in-bud lesions within the three-dimensional space of secondary pulmonary lobule based on radiological/pathological correlation. None of the post-mortem lung specimens showed findings of lung parenchymal lymphatics involvement.

In vitro propagation of Vitis vinifera L. cv. 'Monastrell'

Background: A protocol for the micropropagation of the grape (Vitis vinifera L.) cultivar 'Monastrell' was developed. Initial plant material was obtained from the sanitary selection of grapevine plants performed by real-time RT-PCR to confirm the absence of Grapevine fanleaf virus, Arabis mosaic virus, Grapevine leafroll-associated virus 1, Grapevine leafroll-associated virus 3, and Grapevine fleck virus. Results: The effects of the salt composition (comparing Lloyd and McCown woody plant medium and Murashige and Skoog medium 1/2 macronutrients) and the growth regulator benzylaminopurine (BAP), at 0 and 8.9 µM, on plant propagation were evaluated using nodes as explants. The most efficient procedure consisted of bud induction in the medium with Lloyd and McCown woody plant salts and 8.9 µM BAP for 30 d along with elongation in cytokinin-free medium for 60 d, which gave 22 nodes/explant (174 plants/initial plant). A second cycle of propagation in a medium without BAP for another 60 d could give approximately 10,000 nodes, which can be obtained after an additional 2 months of culture. All plants acclimatized after the second cycle of multiplication were successfully transferred to soil. Conclusion: We developed an optimal protocol for V. vinifera cv. 'Monastrell' micropropagation, the first described for this cultivar.

Effect of irrigation at critical stages on the phenology of flowering and fruiting of the cactus Opuntia spp

Abstract This paper briefly reports some effects of irrigations at two critical periods on the phenology of three varieties of cactus pear cultivated in Agadir area: the spineless varieties Aissa and Moussa and the thorny one Achefri. In the first year experiments (2010-2011) treatments of irrigation used were: (T1) 0 mm, (T2) 30 mm during flowering and 30 mm during fruit enlargement and (T3) 30 mm only during fruit enlargement. In the 2nd year experiments, irrigation treatments were: (T1) 0 mm, (T2) 60 mm during flowering and 60 mm during fruit enlargement and (T3) 60 mm only during fruit enlargement. Treatments of irrigation were applied between mid-April and mid-June in the 1st year experiments and in February and May in the 2nd year experiments. Results of the first year experiments showed that the emission of buds was higher in the thorny variety than in the spineless ones (more than 6 emitted buds/cladode vs less than 4.5 in the spineless varieties). In the second year, irrigation increased the emission of buds in the three varieties (more than 7 emitted buds/cladode for each T2 and T3 of all varieties vs not more than 5 for T1) and the duration of the flowering phase of these varieties. However, irrigation did not modify the proportions of fruits reaching commercial maturity during the early or the late period of maturation.

Resumo Este artigo relatou brevemente alguns efeitos das irrigações em dois períodos críticos na fenologia de três variedades de Opuntia cultivadas na área de Agadir: as variedades sem espinhos 'Aissa' e 'Moussa' e um espinhoso 'Achefri'. No primeiro ano (2010-2011) os seguintes tratamentos do experimento da irrigação foram usados: (T1) 0 milímetros, (T2) 30 milímetros durante a florescência e 30 milímetros durante a ampliação do fruto e (T3) 30 milímetros somente durante a ampliação do fruto. No segundo ano das experiências, os tratamentos da irrigação foram: (T1, 0 milímetros), (T2) 60 milímetros durante a florescência e 60 milímetros durante a ampliação do fruto e (T3) 60 milímetros somente durante a ampliação do fruto. Os tratamentos da irrigação foram aplicados entre a metade de abril e metade de junho nas experiências do primeira ano e em fevereiro e maio nas experiências do segundo ano. Os resultados das experiências do primeiro ano mostraram que a emissão de brotos foi maior na variedade espinhosa do que no rufia (mais de 6 emitida gomos/cladode vs inferior a 4,5 no rufia variedades). No segundo ano, a irrigação aumentou a emissão dos botões nas três variedades (mais de 7 botões emitidos/cladÓdio para cada T2 e T3 de todas as variedades não mais que 5 para o T1) e a duração da fase de florescência destas variedades. Contudo, a irrigação não alterou as proporções de frutos que alcançaram a maturidade comercial durante o período adiantado ou atrasado de maturação.

The application of cytokinesis-block method to analyze cytomics indicators in public health field / 中国医学装备

In recent years, cytokinesis-block method was used to analyze cytomics indicators including micronucleus, nuclear bridge, nuclear bud, nuclear division index, cell apoptosis and cell necrosis. In public health, it has become the common method to explore the impacts of different population structure, environment and occupational exposure for genomic instability, chromosome breakage, chromosome loss and cell proliferation. This article reviews and discusses the application of using cytokinesis block method to analyze cytomics indicators in public health field.

Effects of ginseng flower bud on bone metabolism in rats induced by D-galactose / 中国药理学通报

Aim To investigate the skeletal effects of ginseng flower bud(GF)on osteopenia induced by D-galactose using histomorphometry and biomechanical properties.Methods Fifty three-month-old male Sprague-Dawley rats were randomly divided into five groups.Rats in NS group(NS)were treated with NS(5 mL·kg-1·d-1)by subcutaneous injection and daily oral gavage with vehicle as control.Rats in the other four groups were given D-galactose at the dose of 100 mg·kg-1·d-1 by subcutaneous injection.Solvent control was performed between NS and DG: gastric irrigation with distilled water of 10 mL·kg-1·d-1.Other groups were: CP was gastric irrigated with integrated medicine(stanozolol 0.54 mg·kg-1·d-1+piracetam 432 mg·kg-1·d-1),GF(L)with ginseng flower bud of 0.486 g·kg-1·d-1 and GF(H)with ginseng flower bud of 2.43 g·kg-1·d-1 for 14 weeks.The longitudinal proximal tibial metaphyseal(PTM),the fifth lumbar vertebral body(LVB)and tibial shaft(Tx)sections were performed undecalcifiedly and used for bone histomorphometric analysis.858 Mini Bionix materials testing system was used to analyze the biomechanic properties of right femur via three-point bending test.The left femur was dried and assimilated,whose bone calcium(Ca),phosphate(P)content and bone hydroxyproline content were tested.Results Compared with D-glagatose group,in PTM of D-galactose treated rats,the％Tb.Ar was increased both in GF(L)and GF(H)treated groups.While the Tb.Sp was decreased.％Oc.S.Pm and Oc.N/mm decreased in GF(L),and those in GF(H)were decreased as well.In Tx,％Ct.Ar was raised,while％Ma.Ar was decreased in GF(L)and GF(H).The elastic load of femur was increased.Conclusions Compared with DG group,there are significant differences in bone histomorphometry of Tx and PTM in all doses of GF,but no significant changes are detected in hydroxyproline,Ca,and P content of femur.

Cloning of MADS-box gene AGL15 from Lonicera macranthoides and analysis on its bioinformatics and expression / 中草药

Objective: To clone the MADS-box gene denoted AGL15 from total RNA of Lonicera macranthoides based on previous transcriptome sequencing data and analyze the bioinformatics and expression of the gene. Methods: The gene containing intact open reading frame (ORF) was cloned by reverse transcription-PCR (RT-PCR) and rapid amplification of cDNA ends (RACE). The similarity comparison and homology analysis on the sequence were carried out using bioinformatic method, the coding protein was predicted and the physicochemical properties were analyzed. The expression of the gene in different locations of L. macranthoides was determined by semiquantitative PCR using gene-specific primers. Results: The Lm-AGL15 gene, containing a 795 bp ORF that encoded 264 amino acids, was cloned. The deduced protein sequence had the most similarity to the AGL15 in Vitis vinifera and exhibited two conserved motifs (MADS and K-BOX). Without transmembrane domain, Lm-AGL15 was located in cytoplasm and expressed only in each part. Conclusion: For the first time from L. macranthoides cloning could prolong the period of bud of gene, analysis of the gene expression in different parts of L. macranthoides which would provide a reference for the study of prolonging L. macranthoides bud stage.

Proliferation and organ differentiation of petiole-derived callus in Asarum heterotropoides / 中草药

Objective: To screen the optimum conditions of proliferation, regenerated bud differentiation, and adventitious roots induction of Asarum heterotropoides petiole-derived callus so as to establish the effective system for callus proliferation and regeneration. Methods: Petiole-derived calli were subcultured in the proliferation medium supplemented with different concentration of 6-BA and NAA. Then green and compact calli were chosen to culture in differentiation medium with different exogenous hormones for the induction of regeneration bud and adventitious roots. After being acclimated, regenerated plants were transplanted. Results: The effective callus proliferation medium was 1/2 MS supplemented with 0.40 mg/L 6-BA and 0.10 mg/L NAA, then 1/2 MS medium supplemented with 0.4 mg/L 6-BA and 0.05 mg/L NAA was suitable for regenerated bud differentiation while regenerated buds grew better in 1/2 MS medium without any hormone. The medium of 1/4 MS supplemented with 0.25 mg/L IBA was best for the induction of adventitious roots. Conclusion: The optimum culture conditions of callus proliferation and differentiation in A. heterotropoides are determined to establish the regeneration system and obtain the regenerated plantlet successfully.

Screening of reference genes in Lonicera macranthoides and spatio-temporal expression analysis of LmAGL15 in Mads-box family / 中草药

Objective: To screen the reference genes of Lonicera macranthoides for gene expression analysis and to study the spatio-temporal expression characteristics of LmAGL15 which was a member of Mads-Box family. Methods: In this study, 18 S rRNA, Ubiquilin, Actin and Efl-β of L. macranthoides were cloned and the stabilities of the four housekeeping genes were evaluated in different positions (leaves, stems, and buds) and different periods of bud development. In addition, the spatio-temporal expression of LmAGL15 gene was analyzed. Results: 18 S rRNA was the most suitable reference gene for spatio-temporal expression analysis in L. macranthoides; The relative expression of LmAGL15 was low in leaves and stems, and that in buds was higher. Conclusion: 18 S rRNA is the most suitable reference gene in L. macranthoides. The relative expression of LmAGL15 changes significantly in leaves, stems, and buds.

Study on lateral buds induction and culture of Huperzia serrata / 中草药

Objective: Due to the low reproductive capacity and scarce resources of Huperzia serrata, and in vitro culture of H. serrata has not been really successful, we wish to make a good progress in rapid propagation in this study. Methods: Living shoot tips were used as the materials, after a variety of surface sterilization, malachite green and AAS were used to eliminate the endophytic fungi of the seedling. Then, the aseptic seedlings were transplanted to the medium with 1/2 MS + 25 g/L sucrose to be cultured. After the new dichotomous branching occurred, and the branches grew to 4-5 cm high, we cut it and subcultured for the new dichotomous branching. Results: After four times of subculture of dichotomous branching, the aseptic seedlings got the different number of proliferating lateral buds, which came from the middle or top of the stem. Comparing the structure of lateral buds with the gemma, we found it was significantly different from the gemma. The number of lateral buds from different individuals was different, the most lateral buds could get to 14 in one seedling, some lateral bud generating point can produce two or more lateral buds. After removing the apical dominance of stem tip, the lateral buds in the middle of the stem grew faster. The surviving rate of different height lateral buds was different when they separated from the stock plant. Lateral buds which beyond 5 mm, the surviving rate could get 90%, while cultured in medium of 1/2 MS + 30 g/L sucrose, after 120 d, it grew into seedling about 2-3 cm high, lateral buds less 5 mm had high mortality with the same cultured medium. Conclusion: Though in vitro culture, H. serrata can break the reproductive way by which the leaf axil could generate spore only but no lateral bud naturally, and can produce the multiple lateral bud proliferation. The lateral buds can easily produce adventitious roots by stripping, which could provide an important view for rapid reproduction of H. serrata.

Study on volatile components of Lonicerae Japonicae Flos in bud stage extended type / 中国中药杂志

Volatile components of Lonicerae Japonicae Flos in bud stage extended type Beihua 1 were determined by the headspace solid-phase micro-extraction, compared with traditional cultivar Damaohua. There are fifty-two volatile compounds were identified and the relative content of the volatiles was calculated by the area normalization method. Thirty-nine compounds were found in Beihua 1, whereas thirty-three components in Damaohua. Total twenty identical compounds existed in Beihua 1 and Damaohua. The contents of alcohols and hydrocarbons of Beihua 1 were higher significantly than that of Damaohua, while significantly lower than that of Damaohua in ketones content. Besides, twenty components were only detected in Beihua 1, such as methyl nicotinate, hexadecanoic acid, methyl ester,acetophenone, nonanoic acid.