Phloroglucinol Research Articles

In vitro propagation via organogenesis and embryogenesis of Cyrtanthus mackenii: a valuable threatened medicinal plant

Efficient and simple, organogenesis (direct and indirect) and somatic embryogenesis (cell suspension) systems were developed for in vitro propagation of Cyrtanthus mackenii, a valuable economic plant from leaf explants cultured on Murashige and Skoog (MS) medium supplemented with various concentrations and combinations of sucrose, plant growth regulators (PGRs), glutamine, phloroglucinol (PG) and 6-(2-hydroxy-3-methylbenzylamino) purine (PI55). MS medium solidified with 8 g L−1 agar (MSS) containing 40 g L−1 sucrose, 10 µM picloram, 2.5 µM benzyladenine (BA) and 20 µM glutamine produced a higher number of shoots from white nodular callus. This was however, not significantly different to direct shoot regeneration on media containing 10 µM picloram, 2.5 µM BA and a reduced concentration of sucrose and glutamine. The regenerated shoots were rooted best with MSS medium incorporating 10 µM PG. The number of somatic embryos (SEs) were significantly higher using liquid MS medium containing 30 g L−1 sucrose, 0.5 µM picloram, 1 µM thidiazuron or BA and 3 µM glutamine or gibberellic acid. The embryos were germinated in PGR-free MSS medium. All plantlets were successfully acclimatized in the greenhouse. Histological studies confirmed the different developmental stages and bipolar structure of SE. The organogenesis and somatic embryogenesis protocols provides a system for large scale propagation and germplasm conservation. Developed protocols can be used for clonal production and pharmacological and genetic transformation studies.

Influence of biostimulants-seed-priming on Ceratotheca triloba germination and seedling growth under low temperatures, low osmotic potential and salinity stress

Extreme temperatures, drought and salinity stress adversely affect seed germination and seedling growth in crop species. Seed priming has been recognized as an indispensable technique in the production of stress-tolerant plants. Seed priming increases seed water content, improves protein synthesis using mRNA and DNA and repair mitochondria in seeds prior to germination. The current study aimed to determine the role of biostimulants-seed-priming during germination and seedling growth of Ceratotheca triloba (Bernh.) Hook.f. (an indigenous African leafy vegetable) under low temperature, low osmotic potential and salinity stress conditions. Ceratotheca triloba seeds were primed with biostimulants [smoke-water (SW), synthesized smoke-compound karrikinolide (KAR1), Kelpak® (commercial seaweed extract), phloroglucinol (PG) and distilled water (control)] for 48h at 25°C. Thereafter, primed seeds were germinated at low temperatures, low osmotic potential and high NaCl concentrations. Low temperature (10°C) completely inhibited seed germination. However, temperature shift to 15°C improved germination. Smoke-water and KAR1 enhanced seed germination with SW improving seedling growth under different stress conditions. Furthermore, priming seeds with Kelpak® stimulated percentage germination, while PG and the control treatment improved seedling growth at different PEG and NaCl concentrations. Generally, high concentrations of PEG and NaCl brought about detrimental effects on seed germination and seedling growth. Findings from this study show the potential role of seed priming with biostimulants in the alleviation of abiotic stress conditions during seed germination and seedling growth in C. triloba plants.

Advantage of childbirth with ball-assisted free body position

Objective To evaluate the clinical effect and safety of intravenous administration of phloroglucinol combined with delivery of ball-assisted free position in parturient women during labor. Methods One hundred and fifty-two primiparous women who were treated by vaginal delivery from June 2015 to August 2016 in Beijing Navy General Hospital were selected as the study subjects.They were given 80 mg of phloroglucinol diluted in saline injection intravenous injection.Observation group were treated with childbirth.Control group treated with traditional supine or lateral body. Results The rate of descending of the observed group was longer than that of the control group((1.09±0.22) cm/h vs.(0.83±0.15) cm/h), the first labor time, the second labor time and the total labor time of the observation group were shorter than the control group((9.57±1.71) h vs.(12.13±2.64) h, (35.75±8.68) min vs.(61.21±10.03) min, (11.34±1.96) h vs.(14.66±2.70) h), the difference between the two groups was statistically significant(t=3.611, 4.204, 3.307, 3.456, P 0.05). The percentage of vaginal delivery in the observation group was 89.47%(68/76), significantly higher than that in the control group(73.68%(56/76)), the difference was statistically significant(χ2=4.514, P<0.05). The incidence of vaginal midwifery was 1.32%(1/76) and that of cesarean section was 9.21%(7/76) of observation group, significantly lower than that in control group(5.26%(4/76), 21.05% (16/76)), the difference was statistically significant(χ2=5.323, 4.376, P<0.05). Observation group 24 h postpartum hemorrhage was (273.12±58.23) ml, significantly less than the control group((385.42±70.13) ml), the difference between the two groups was statistically significant(t=3.744, P<0.05). The observation group had postpartum hemorrhage of 1.32%(1/76), urinary retention of 2.63%(2/76), and the percentage of neonatal asphyxia of 2.63%(2/76), were lower than that of the control group(5.26%(4/76), 10.53%(8/76), 9.21%(7/76)), the difference between the two groups was statistically significant(χ2=4.103, 4.925, 4.421, P<0.05). Conclusion The delivery of ball-assisted free-body delivery in active stage of delivery can promote natural childbirth.The medical staff should update the concept of delivery and raise awareness to encourage the use of phloroglucinol combined with free-body delivery. Key words: Active phase; Phloroglucinol; Delivery ball; Free postural delivery

In vitro plant regeneration and alleviation of physiological disorders in Scadoxus puniceus

The frequent overharvesting of slow-growing bulbs has led to a severe reduction in wild plant populations. In vitro propagation techniques have been useful in reducing the stress placed on wild plant populations. The techniques involved in in vitro propagation of the Amaryllidaceae are often challenging. The current study evaluated the role of phloroglucinol (PG), gallic acid (GCA; an inhibitor of phenylalanine ammonia lyase) and photoperiod on the development of bulblets, reduction of phenolic exudation and hyperhydricity in Scadoxus puniceus grown in vitro. The combination of PG and naphthaleneacetic acid (NAA) significantly increased the number of bulblets that developed on leaf explants. GCA (1, 5 and 10μM) and 5μM benzyladenine (BA) significantly improved the number of bulblets produced on leaf explants and plant development. The longest shoots (66.0±9.03mm) were produced at 10μM GCA. In addition, red pigmentation characteristic of phenolic exudation in the media were reduced with treatments of PG and GCA as well as a 24h dark photoperiod. Hyperhydricity was alleviated with the application of meta-topolins. The shoots of regenerated bulb halves developed to a length of 180mm under a 24h dark photoperiod. The current study is the first to describe an in vitro propagation protocol for S. puniceus by limiting the effect of phenolic exudation thus providing a better culture condition and it provides a better conservation strategy for this over-utilised species.

Ultrastructure of somatic embryo development and plant propagation for Lachenalia montana

Embryogenesis for plant propagation by cell suspension culture was achieved for the first time using friable embryogenic callus (FEC) from leaf explants of Lachenalia montana Schltr. ex W.F. Barker. FEC was established with solid (8gL−1 agar) MS (Murashige and Skoog, 1962) medium containing various concentrations and combinations of sucrose and plant growth regulators (PGRs). The somatic embryos (SEs) were developed from FEC in liquid MS (MSL) medium with or without PGRs. A higher number of SEs of different developmental stages (26.0–19.4, globular to cotyledonary-stages respectively) were obtained on MSL medium with 0.5μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 1μM thidiazuron (TDZ). The germination frequency (33.7%) was highest in MSL medium containing 1μM 2,4-D and 2μM TDZ. The different stages of SEs germinated (92%) on solid full-strength MS medium containing 15gL−1 sucrose and 10μM phloroglucinol (PG). The plantlets were successfully acclimatized in the greenhouse. Developmental stages and features of embryoids were confirmed by histological and ultrastructural studies using light and transmission electron microscopy (TEM). Many mitochondria, lipid bodies together with starch grains, chloroplasts, Golgi apparatuses, vacuoles and nuclei were detected. The system developed offers controlled large-scale clonal propagation which ensures germplasm conservation, confirms viability of embryogenesis by ultrastructural studies and provides a system for genetic transformation studies for horticultural and ornamental improvement.

Improving the production of acetyl-CoA-derived chemicals in Escherichia coli BL21(DE3) through iclR and arcA deletion

BackgroundAcetyl-CoA-derived chemicals are suitable for multiple applications in many industries. The bio-production of these chemicals has become imperative owing to the economic and environmental problems. However, acetate overflow is the major drawback for acetyl-CoA-derived chemicals production. Approaches for overcoming acetate overflow may be beneficial for the production of acetyl-CoA-derived chemicals.ResultsIn this study, a transcriptional regulator iclR was knocked out in E.coli BL21(DE3) to overcome acetate overflow and improve the chemicals production. Two important acetyl-CoA-derived chemicals, phloroglucinol (PG) and 3-hydroxypropionate (3HP) were used to evaluate it. It is revealed that knockout of iclR significantly increased expressions of aceBAK operon. The cell yields and glucose utilization efficiencies were higher than those of control strains. The acetate concentrations were decreased by more than 50% and the productions of PG and 3HP were increased more than twice in iclR mutants. The effects of iclR knockout on cell physiology, cell metabolism and production of acetyl-CoA-derived chemicals were similar to those of arcA knockout in our previous study. However, the arcA-iclR double mutants couldn’t gain higher productions of PG and 3HP. The mechanisms are unclear and needed to be resolved in future.ConclusionsKnockout of iclR significantly increased gene expression of aceBAK operon and concomitantly activated glyoxylate pathway. This genetic modification may be a good way to overcome acetate overflow, and improve the production of a wide range of acetyl-CoA-derived chemicals.

Enhancing plant regeneration of Lachenalia viridiflora, a critically endangered ornamental geophyte with high floricultural potential

Lachenalia viridiflora is a critically endangered bulbous plant with high potential on the international floriculture market. In the present study, an efficient protocol for in vitro plantlet regeneration through somatic embryogenesis was developed. Embryogenic callus was established from leaf explants cultured on Murashige and Skoog (MS) basal medium supplemented with various concentrations and combinations of picloram and thidiazuron (TDZ). A high number of somatic embryos (28.5±1.49) at different developmental stages of somatic embryos (SEs: globular embryos, juvenile embryo and cotyledon with bipolar characteristics) was obtained on Murashige and Skoog (1962) medium with 2.5μM picloram, and 1.0μM TDZ. Histological and scanning electron microscope (SEM) analyses confirmed the presence of somatic embryos. Mature somatic embryos germinated and developed into plantlets after 6 weeks on full strength MS medium. High plant regeneration frequency (91.1%) was achieved on full-strength MS medium supplemented with 5μM phloroglucinol (PG). Well-developed healthy plantlets were successfully acclimatized in the greenhouse with a survival rate of 90%. The result of this study is beneficial in the mass propagation of high-quality L. viridiflora plants for the commercial horticultural market and also provides a platform for future genetic transformation studies.

Supramolecular hydrogen-bonding patterns of co-crystals containing the active pharmaceutical ingredient (API) phloroglucinol and N-heterocycles.

The active pharmaceutical ingredient phloroglucinol (PHL) has been taken as an illustrative molecule to explore the intermolecular interactions which can be established with other molecular entities to build PHL pharmaceutical co-crystals. The crystal structures of five newly synthesized co-crystals are reported, where PHL is crystallized with N-heterocycles, namely 2-hydroxy-6-methylpyridine (1), 2,4-dimethyl-6-hydroxypyrimidine (2), 4-phenylpyridine (3), 2-hydroxypyridine (4) and 2,3,5,6-tetramethylpyrazine (5). The structural characteristics of these co-crystals, as far as the hydrogen-bonding networks and the crystalline architectures are concerned, are strongly dependent on the chemical features of the coformer molecules, as well as on their size and shape. A detailed analysis of the intermolecular interactions established in all the PHL co-crystals of known structures has allowed the recognition of some regularities in the packing modes that can be useful in the design of new supramolecular adducts forming predictable structural motifs.

Enolate-Forming Phloretin Pharmacophores: Hepatoprotection in an Experimental Model of Drug-Induced Toxicity.

Drug-induced toxicity is often mediated by electrophilic metabolites, such as bioactivation of acetaminophen (APAP) to N-acetyl-p-benzoquinone imine (NAPQI). We have shown that APAP hepatotoxicity can be prevented by 2-acetylcyclopentanone (2-ACP). This 1,3-dicarbonyl compound ionizes to form an enolate nucleophile that scavenges NAPQI and other electrophilic intermediates. In this study, we expanded our investigation of enolate-forming compounds to include analyses of the phloretin pharmacophores, 2',4',6'-trihydroxyacetophenone (THA) and phloroglucinol (PG). Studies in a mouse model of APAP overdose showed that THA provided hepatoprotection when given either by intraperitoneal injection or oral administration, whereas PG was hepatoprotective only when given intraperitoneally. Corroborative research characterized the molecular pharmacology (efficacy, potency) of 2-ACP, THA, and PG in APAP-exposed isolated mouse hepatocytes. For comparative purposes, N-acetylcysteine (NAC) cytoprotection was also evaluated. Measurements of multiple cell parameters (e.g., cell viability, mitochondrial membrane depolarization) indicated that THA and, to a lesser extent, PG provided concentration-dependent protection against APAP toxicity, which exceeded that of 2-ACP or NAC. The enolate-forming compounds and NAC truncated ongoing APAP exposure and thereby returned intoxicated hepatocytes toward normal viability. The superior ability of THA to protect is related to multifaceted modes of action that include metal ion chelation, free radical trapping, and scavenging of NAPQI and other soft electrophiles involved in oxidative stress. The rank order of potency for the tested cytoprotectants was consistent with that determined in a parallel mouse model. These data suggest that THA or a derivative might be useful in treating drug-induced toxicities and other conditions that involve electrophile-mediated pathogenesis.

Deletion of arcA increased the production of acetyl-CoA-derived chemicals in recombinant Escherichia coli.

Acetyl-CoA is used to produce many valuable metabolites in Escherichia coli. However, acetate overflow is a major shortcoming. Knockout of the global regulator gene, arcA, may solve this problem. The arcA gene of E. coli BL21(DE3) was knocked out, and the production of phloroglucinol (PG) and 3-hydroxypropionate (3HP), both derived from acetyl-CoA, were used to evaluate its effect. The arcA mutants had higher cell yields and higher glucose utilization efficiencies than the corresponding control strains, and the productions of PG and 3HP were 0.92 g/l and 0.27 g/l, respectively; more than twice that of the control strains. Furthermore, arcA knockout also showed significant repression on formation of acetate, the major byproduct in fermentation. Acetate concentrations were decreased 69.4 % and 87 % by arcA knockout during the production of PG and 3HP, respectively. The arcA gene knockout is a solution to acetate overflow and may improve production of a wide range of acetyl-CoA-derived metabolites.

In vitro propagation and biochemical changes in Aloe pruinosa

Aloe pruinosa is a vulnerable traditional medicinal plant extensively used in South Africa. Efficient in vitro mass propagation and analysis of biochemical changes of the species was established to counter extinction threats, for commercial cultivation and to understand the effect of oxidative enzymes and phenolic metabolism on oxidative browning in vitro from shoot-tip and leaf explants using various concentrations and combinations of plant growth regulators (PGRs), sucrose and phloroglucinol (PG). The highest number of regenerated shoots per shoot-tip explant (15.4±0.9) was obtained on solid Murashige and Skoog (MSS) medium supplemented with a combination of 5μM benzyladenine (BA) and 4μM indole-3-acetic acid (IAA). Shoots developed significantly more roots (8.6±0.7 per shoot) on ½-MSS medium containing 2μM indole-3-butyric acid (IBA) and 40μM PG. The plantlets were successfully acclimatized (100%) ex vitro in a vermiculite-soil mixture (1:1 v/v) in the greenhouse. Somatic embryogenesis with all developmental stages of somatic embryos (SEs: 38–26) were obtained from friable embryogenic callus (FEC) of leaf explants in liquid MS (MSL) containing 0.5μM picloram (Pic), 2μM BA or 1μM thidiazuron (TDZ) and 10μM PG. However, germination and conversion of plantlets from SEs were inhibited as a result of oxidative browning. Polyphenolic enzymes (polyphenol oxidase (PPO) and peroxidase (POD)) activity and total phenolics of different in vitro growth phases (adventitious shoots, regenerated plantlets and calli) with different PGR treatments were determined. The application of PGRs and PG resulted in the control of morphogenic properties, oxidative exudates and browning in A. pruinosa. The study highlights the role of natural PGR (meta-topolin riboside) and PG in reducing the impact of polyphenolic exudation and can be used to improve future in vitro programmes. In addition, micropropagation protocols facilitate commercial and rapid propagation of A. pruinosa for conservation and could satisfy pharmaceutical, cosmetic and food industry demands.

Novel anticancer activity of phloroglucinol against breast cancer stem-like cells

Poor prognosis of breast cancer patients is closely associated with metastasis and relapse. There is substantial evidence supporting that cancer stem-like cells (CSCs) are primarily responsible for relapse in breast cancer after anticancer treatment. However, there is a lack of suitable drugs that target breast cancer stem-like cells (BCSCs). Here, we report that phloroglucinol (PG), a natural phlorotannin component of brown algae, suppresses sphere formation, anchorage-independent colony formation and in vivo tumorigenicity. In line with these observations, treatment with PG also decreased CD44+ cancer cell population as well as expression of CSC regulators such as Sox2, CD44, Oct4, Notch2 and β-catenin. Also, treatment with PG sensitized breast cancer cells to anticancer drugs such as cisplatin, etoposide, and taxol as well as to ionizing radiation. Importantly, PG inhibited KRAS and its downstream PI3K/AKT and RAF-1/ERK signaling pathways that regulate the maintenance of CSCs. Taken together, our findings implicate PG as a good candidate to target BCSCs and to prevent the disease relapse.

The clinical study of different time used phloroglucin on ureteral calculi by extracorporeal shock wave lithotripsy

Objective To discuss the application of different time phloroglucinol for calculi after extracorporeal shock wave lithotripsy eduction. Methods 102 patients with ureteral calculi were chosen as the research objects, who were randomly divided into the observation group and control group, both for extracorporeal shock wave lithotripsy treatment, the control group of gravel for 15 min after intravenous injection of phloroglucinol,the observation group of gravel 15 min before application of phloroglucinol, observed the two groups of rows of stone effect. Results T- test andχ2 test method, the observation group were cured 30 cases(58.82%), compared with the control group(χ2= 7.140,P= 0.007), the difference was statistically significant. Two incidence of adverse reactions and renal colic recurrence rate was relatively(χ2= 0.210,0.101,P= 0.646,0.749), the difference was not statistically significant. Conclusion Extracorporeal shock wave lithotripsy 15 min before application of phloroglucinol can promote calculous eduction, improve the effect of clinical treatment, no increase in adverse reactions, whichis worth popularizing in clinical use. Key words: Extracorporeal shock wave lithotripsy; Ureteral calculi; Phloroglucin

Physiological role of phenolic biostimulants isolated from brown seaweed Ecklonia maxima on plant growth and development.

Eckol, a major phenolic compound isolated from brown seaweed significantly enhanced the bulb size and bioactive compounds in greenhouse-grown Eucomis autumnalis. We investigated the effect of eckol and phloroglucinol (PG) (phenolic compounds) isolated from the brown seaweed, Ecklonia maxima (Osbeck) Papenfuss on the growth, phytochemical and auxin content in Eucomis autumnalis (Mill.) Chitt. The model plant is a popular medicinal species with increasing conservation concern. Eckol and PG were tested at 10(-5), 10(-6) and 10(-7)M using soil drench applications. After 4months, growth parameters, phytochemical and auxin content were recorded. When compared to the control, eckol (10(-6)M) significantly improved bulb size, fresh weight and root production while the application of PG (10(-6)M) significantly increased the bulb numbers. However, both compounds had no significant stimulatory effect on aerial organs. Bioactive phytochemicals such as p-hydroxybenzoic and ferulic acids were significantly increased in eckol (10(-5)M) and PG (10(-6)M) treatments, compared to the control. Aerial (1,357pmol/g DW) and underground (1,474pmol/g DW) parts of eckol-treated (10(-5)M) plants yielded the highest concentration of indole-3-acetic acid. Overall, eckol and PG elicited a significant influence on the growth and physiological response in E. autumnalis. Considering the medicinal importance of E. autumnalis and the increasing strains on its wild populations, these compounds are potential tools to enhance their cultivation and growth.

Kinetic Entrapment of a Hidden Curcumin Cocrystal with Phloroglucinol

The existence of a cocrystal between curcumin (CUR) and phloroglucinol (PHL) was suspected but could not be demonstrated in a recent systematic effort to synthesize novel curcumin cocrystals. We hypothesize that the elusive CUR–PHL cocrystal is a kinetically stable form that can be prepared by trapping it using a fast solvent removal crystallization process. The polarity of crystallization solvent and relative solubility of cocrystal formers in the solvent appear to be critical parameters governing the phase purity of the resulting cocrystal. Organic solvents of higher polarity and in which the cocrystal formers exhibited congruent solubility tended to afford a purer cocrystal phase. Essentially phase-pure CUR–PHL cocrystals were successfully obtained from acetone, and their 1:1 stoichiometry was confirmed by differential scanning calorimetry and solid state nuclear magnetic resonance spectroscopy. Compared with the individual component phases, the cocrystal displayed reduced hygroscopicity and improved t...

Influence of m-trihydroxybenzene on the recurrence rate in children with intussusception hydrostatic enema

Objective To explore the application value of m-trihydroxybenzene in pediatric intussusception in the course of treatment,to observe its curative effect and safety.Methods 200 cases of enema intussusception were selected and randomly divided into two groups by the method of random number table.100 cases in the observation group were given m-trihydroxybenzene injection (2mg/kg) 15 minutes before hydrostatic enema,and reduction of hydrostatic enema.If reduction,continued to give benzene three phenol intravenous drip,once a day.100 cases in the control group used atropine(0.02mg/kg) intramuscular injection before hydrostatic enema,after successful reduction did not continue to use the spasmolytic drug.The success rate,short-term recurrence rate,reduction time and adverse reaction were compared between the two groups.Results The hydrostatic pressure enema reduction success rate of the observation group was 99％,which was significantly higher than 87％ in the control group (x2 =4.38,P ＜ 0.05).The reduction time of the observation group was (7.56 ± 2.54) min,which was significantly shorter than (10.45 ± 3.03) min of the control group (t =-4.285,P ＜ 0.05).The short-term recurrence rate in 3 days of the observation group was 3％,which was significantly lower than 13％ in the control group (x2 =2.79,P ＜0.05).The observation group had no obvious adverse reaction.Conclusion m-Trihydroxybenzene in water enema intussusception reset before and after use is safe and effective,it can not only improve the success rate reduction,but also can significantly reduce the short-term recurrence of intestinal intussusception reduction rate. Key words: Phloroglucinol; Intussusception; Enema; Ultrasonography; Child

Posttranscriptional regulation of 2,4-diacetylphloroglucinol production by GidA and TrmE in Pseudomonas fluorescens 2P24.

Pseudomonas fluorescens 2P24 is a soilborne bacterium that synthesizes and excretes multiple antimicrobial metabolites. The polyketide compound 2,4-diacetylphloroglucinol (2,4-DAPG), synthesized by the phlACBD locus, is its major biocontrol determinant. This study investigated two mutants defective in antagonistic activity against Rhizoctonia solani. Deletion of the gidA (PM701) or trmE (PM702) gene from strain 2P24 completely inhibited the production of 2,4-DAPG and its precursors, monoacetylphloroglucinol (MAPG) and phloroglucinol (PG). The transcription of the phlA gene was not affected, but the translation of the phlA and phlD genes was reduced significantly. Two components of the Gac/Rsm pathway, RsmA and RsmE, were found to be regulated by gidA and trmE, whereas the other components, RsmX, RsmY, and RsmZ, were not. The regulation of 2,4-DAPG production by gidA and trmE, however, was independent of the Gac/Rsm pathway. Both the gidA and trmE mutants were unable to produce PG but could convert PG to MAPG and MAPG to 2,4-DAPG. Overexpression of PhlD in the gidA and trmE mutants could restore the production of PG and 2,4-DAPG. Taken together, these findings suggest that GidA and TrmE are positive regulatory elements that influence the biosynthesis of 2,4-DAPG posttranscriptionally.

In vitro regeneration and conservation of the lentisk (Pistacia lentiscus L.)

Pistacia lentiscus L. (lentisk or mastic tree) is an economically important member of the genus Pistacia due to its valuable mastic resin. Shoot tips and nodal segments were used as explant sources from in vitro-germinated seeds of lentisk. Shoot tips were found to be suitable for multiple shoot formation. Thereafter, the influences of different growth regulators [N6-benzyladenine (BA), gibberellic acid (GA3), naphthalene acetic acid (NAA), or jasmonic acid (JA)] together with various elicitors [silver nitrate (AgNO3) or phloroglucinol (PG)] were assessed to develop efficient micropropagation protocol. Our results showed that the combination of all tested concentrations of GA3 with 1.0 mg/L BA resulted in enhancement of multiple shoot formation. The maximum number of shoots per explant (3.95) was recorded on MS medium containing 1.0 mg/L BA and 0.3 mg/L GA3. In contrast, JA had a negative influence, while AgNO3 had no significant effect on multiple shoot formation. In terms of synthetic seed production, it was possible to encapsulate shoot tips at 4 °C in darkness for up to 6 months with a frequency of 87.5% plant regrowth. In vitro-propagated microshoots, including plantlets derived from synthetic seeds, were transferred to MS medium containing different concentrations (1.0, 2.0, or 4.0 mg/L) of indole butyric acid for rooting and successfully acclimatized to ex vitro conditions. The presented data suggest an efficient in vitro regeneration system and conservation via synthetic seed production for lentisk.

Encapsulation of nodal segments of Cassia angustifolia Vahl. for short-term storage and germplasm exchange

The present study described the encapsulation of nodal segments of Cassia angustifolia Vahl. excised from 1-month-old in vitro raised cultures for short-term conservation and propagation. Various concentrations and combinations of gelling matrix (sodium alginate) and complexing agents (calcium chloride) were tested to prepare uniform beads. The ideal beads were obtained through a combination of 3 % sodium alginate and 100 mM calcium chloride. The maximum conversion response (94 %) of encapsulated beads was obtained in Murashige and Skoog’s medium (MS medium) supplemented with 2.5 μM benzyladenine (BA) and 0.4 μM α-naphthalene acetic acid (NAA) after 6 weeks of culture. The encapsulated and non-encapsulated nodal segments were also stored at 4 °C for different time periods (0, 1, 2, 4, 6 and 8 weeks). The regenerated microshoots were best rooted in optimized rooting medium that comprised half-strength MS + 1.0 μM indole-3-butyric acid (IBA) + 5.0 μM phloroglucinol (PG) for the production of complete plantlets. The regenerated plantlets were successfully hardened and acclimatized in natural conditions with 70 % survival rate.

Clinical application of antispasmodic drugs in ureteroscopic lithotripsy

Objective To explore clinical value of preoperative use of ureteral antispasmodic drugs in ureteroscopy lithotripsy.Methods 252 patients with ureteral calculus were randomly divided into five groups:Group A with 50 patients,as the control group; Group B,51 patients,were treated with Phloroglucinol (80 mg,ivgtt,once daily); Group C,50 patients,were treated with tamsulosin (0.4 mg,po,once daily),Group D,49 patients,were treated with 654-2 (10 mg,ivgtt,twice daily); Group E,52 patients,were treated with progesterone (40 mg,im,once daily).All groups were given anti-inflammatory treatment preoperatively.8.0/9.8 F rigid ureteroscope was used for ureteroscopic lithotomy or lithotripsy 3 days after antispasmodic treatment.Results The operation was successful in all cases.The overall stone-free rate was 98.8％(249/252).In the operation,there was statistical difference in using reteroscope under direct vision between group A and E (P ＜ 0.05).There were statistical differences in postoperative complication incidence between group A and B,C,D,E (P ＜ 0.05).After 3-18 months' (average 9 months) follow-up period,there was no recurrence and no ureterostenosis was found.Conclusions Routine preoperative use of antispasmodic drugs can relax ureteral smooth muscle,make the ureter relaxed and diastolic,and significantly lower the incidence of intraoperative ureteral injuries.The antispasmodic effect of progesterone is the best among several antispasmodic drugs,with few side effects. Key words: Anticonvulsive drug; Ureteral injury; Ureteroscope