Simultaneous determination of three constituents in Chunyang Zhengqi capsules by HPLC / 药学实践杂志

Objective To establish method for simultaneous determination of hesperidin, cinnamaldehyde and eugenol in Chunyang Zhengqi capsules by high performance liquid chromatography. Methods The column was Agilent PorosheⅡ 120 EC-C18 (4.6 mm×150 mm, 4 μm). The mobile phase was acetonitrile-water with gradient elution. The column temperature was 35℃. The flow rate was 1.0 ml/min, and the detection wavelength was 284 nm. Results The methodological verification showed that hesperidin, cinnamaldehyde and eugenol had a good linearity (r≥0.999 9). The precisions were less than 2.0%. The average recovery was between 98.0% and 101.9%. The stability and repeatability of RSD were also less than 3.0%, which met the requirements of method validation. Conclusion The method is simple, stable, reproducible and accurate, which could be used to the quality control of Chunyang Zhengqi capsules.

POSTOPERATIVE PAIN INCIDENCE AND INTENSITY FOLLOWING ROOT CANAL OBTURATION WITH BIOCERAMIC AND OTHER SEALERS: A SYSTEMATIC REVIEW WITH META-ANALYSIS

Aims: To investigate whether bioceramicsealers induce a lower incidence and intensity of postoperative pain compared to other sealers. Materials and Methods: Six electronic databases were searched for studies published up to April 2022, following the PICOS strategy: (P) adult patients undergoing root canal treatment or retreatment; (I) root canal filling using bioceramic sealer; (C) root canal filling using other types of sealers; (O) Primary: postoperative pain incidence and/or intensity; Secondary: number of medication intake; (S) randomizedclinical trials. Risk of bias assessment was performed with the revised Cochrane risk of bias tools for randomized trials (RoB 2). Overall certainty of evidence was assessed through the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) tool. Results: Ten studies were included. Eight studies had a low risk of bias, and two had some concerns risk. Meta-analyses showed no differences regarding postoperative pain intensity and incidence between bioceramic sealers and AH Plus. Number of medication intake seemed to be associated to the preoperative diagnosis. Zinc oxide-eugenol sealer demonstrated an intense postoperative pain compared to bioceramic sealers and AH Plus. GRADE analysis showed a low certainty of evidence for all outcomes. Conclusions: There seem to be no differences between bioceramic sealers and AH Plus regarding postoperative pain intensity and incidence. Number of medication intake seem to be associated to the preoperative diagnosis. Zinc oxide-eugenol evoked a more pronounced postoperative pain.

Characteristic features and comparative analysis of essential oil composition of selected genus of Ocimum sanctum L. through GC-MS

Background & objectives: The main aim of this research is to provide literature on the Ocimum plant, and to know the significance of the Ocimum species carried out by pharmacognostic study and experimental design for GC-MS. Ocimum genus are very important for their therapeutic potential among the most important aromatic herbs. Methods: Extreme attention has been put on literature reports in which the utilization of tulsi and their pharmacognostic study has been done by performing morphological and microscopic leaf experimental design and by using essential oil through the GC-MS instrumentation method. Results:The utilization of these characteristics would be important for the drug discovery scientist to develop a specific formulation of the crude drug, which will be a magical therapeutic agent in the future, with many advantages. GC-MS chromatogram of Ocimum sanctum, Ocimum canum, and Ocimum gratissimum oil showed major peaks and has been identified after comparison of the mass spectra with the NIST library, indicating the presence of three phytocomponents. From the results, the GC-MS study suggested that anethole which is well reported antimicrobial compound is more in O. canum (2.66%) in comparison to O. sanctum (1,28%) but absent in O. gratissimum. The results indicated that the antimicrobial activity is more in O. canum due to the presence ofa high amount of anethole in comparison to O. gratissimum and O. sanctum. Interpretation & conclusion: The result revealed that O. canum has a microscopic character that can be identified by the characteristic GC MS analysis of extracts to distinguish between different species of the ocimum plant.

The effectiveness of EDTA 17% as a cleaning solution for the fiber post space after filling with cements

Aim: To evaluate the resistance of the union between a glass fiber post and radicular dentine after cleaning the root with 17% EDTA and filling with different endodontic cements. Methods: Forty uniradicular bovine incisors were removed to obtain root lengths of 18 mm. Endodontic treatment was performed on all roots using different filling cements (zinc oxide and eugenol-based, OZE; cement based on epoxy resin, AH) and cleaning solutions (saline, SA or EDTA), which made it possible to obtain four groups: OZESA, OZEEDTA, AHSA and AHEDTA. Subsequently, 12 mm of filling material was removed from the roots, and they were prepared to receive fiber posts luted with resin cement. To execute the mechanical cycles (2x106 cycles, 90 N, 4 Hz), coronal reconstruction was performed with a silicon matrix. The roots were then sliced (2-mm thick) to perform the push-out test. The results were analyzed using analysis of variance (one factor and two factors) and Tukey's test (α=0,05). Results: Bond strength (Mpa) was significantly higher for OZEEDTA (9,18) and AHEDTA (8,70) than for OZESA (6,06) AHSA (8,7). OZEEDTA also presented the highest values in the cervical region (15,18) but was significantly lower in the apical region (2,99). However, AHEDTA had a homogeneous bond strength in all thirds. Conclusion: Regardless of the endodontic cement used, EDTA was used as an irrigating solution, culminating in a higher bond strength between the glass fiber post and dentin

Eugenol and its liposome-based nano carrier reduce anxiety by inhibiting glyoxylase-1 expression in mice / Eugenol e seu nanocarreador à base de lipossoma reduzem a ansiedade ao inibir a expressão de glioxalase 1 em camundongos

The most common form of psycho-social dysfunction is anxiety with depression being related closely without any age bar. They are present with combined state of sadness, confusion, stress, fear etc. Glyoxalase system contains enzyme named glyoxalase 1 (GLO1).It is a metabolic pathway which detoxifies alpha-oxo-aldehydes, particularly methylglyoxal (MG). Methylglyoxal is mainly made by the breakdown of the glycolytic intermediates, glyceraldehyde-3-phosphates and dihydroxyacetone phosphate. Glyoxylase-1 expression is also related with anxiety behavior. A casual role or GLO-1 in anxiety behavior by using viral vectors for over expression in the anterior cingulate cortex was found and it was found that local GLO-1 over expression increased anxiety behavior. The present study deals with the molecular mechanism of protective activity of eugenol against anxiolytic disorder. A pre-clinical animal study was performed on 42 BALB/c mice. Animals were given stress through conventional restrain model. The mRNA expression of GLO-1 was analyzed by real time RT-PCR. Moreover, the GLO-1 protein expression was also examined by immunohistochemistry in whole brain and mean density was calculated. The mRNA and protein expressions were found to be increased in animals given anxiety as compared to the normal control. Whereas, the expressions were decreased in the animals treated with eugenol and its liposome-based nanocarriers in a dose dependent manner. However, the results were better in animals treated with nanocarriers as compared to the compound alone. It is concluded that the eugenol and its liposome-based nanocarriers exert anxiolytic activity by down-regulating GLO-1 protein expression in mice.

A forma mais comum de disfunção psicossocial é a ansiedade intimamente relacionada com a depressão, sem qualquer barreira de idade. Elas estão presentes em um estado combinado de tristeza, confusão, estresse, medo etc. O sistema de glioxalase contém uma enzima chamada glioxalase 1 (GLO1). É uma via metabólica que desintoxica alfa-oxo-aldeídos, particularmente metilglioxal (MG). O metilglioxal é produzido principalmente pela quebra dos intermediários glicolíticos, gliceraldeído-3-fosfatos e fosfato de diidroxiacetona. A expressão da glioxalase 1 também está relacionada ao comportamento de ansiedade. Um papel casual ou GLO1 no comportamento de ansiedade usando vetores virais para superexpressão no córtex cingulado anterior foi encontrado e descobriu-se que a superexpressão local de GLO1 aumentava o comportamento de ansiedade. O presente estudo trata do mecanismo molecular da atividade protetora do eugenol contra o transtorno ansiolítico. Um estudo pré-clínico em animais foi realizado em 42 camundongos BALB / c. Os animais foram submetidos ao estresse por meio do modelo de contenção convencional. A expressão de mRNA de GLO1 foi analisada por RT-PCR em tempo real. Além disso, a expressão da proteína GLO1 também foi examinada por imuno-histoquímica em todo o cérebro e a densidade média foi calculada. Verificou-se que as expressões de mRNA e proteínas estavam aumentadas em animais que receberam ansiedade em comparação com o controle normal. Considerando que as expressões foram diminuídas nos animais tratados com eugenol e seus nanocarreadores baseados em lipossomas de forma dependente da dose. No entanto, os resultados foram melhores em animais tratados com nanocarreadores em comparação com o composto sozinho. Conclui-se que o eugenol e seus nanocarreadores baseados em lipossomas exercem atividade ansiolítica por regulação negativa da expressão da proteína GLO1 em camundongos.

Eugenol and its liposome-based nano carrier reduce anxiety by inhibiting glyoxylase-1 expression in mice

Abstract The most common form of psycho-social dysfunction is anxiety with depression being related closely without any age bar. They are present with combined state of sadness, confusion, stress, fear etc. Glyoxalase system contains enzyme named glyoxalase 1 (GLO1).It is a metabolic pathway which detoxifies alpha-oxo-aldehydes, particularly methylglyoxal (MG). Methylglyoxal is mainly made by the breakdown of the glycolytic intermediates, glyceraldehyde-3-phosphates and dihydroxyacetone phosphate. Glyoxylase-1 expression is also related with anxiety behavior. A casual role or GLO-1 in anxiety behavior by using viral vectors for over expression in the anterior cingulate cortex was found and it was found that local GLO-1 over expression increased anxiety behavior. The present study deals with the molecular mechanism of protective activity of eugenol against anxiolytic disorder. A pre-clinical animal study was performed on 42 BALB/c mice. Animals were given stress through conventional restrain model. The mRNA expression of GLO-1 was analyzed by real time RT-PCR. Moreover, the GLO-1 protein expression was also examined by immunohistochemistry in whole brain and mean density was calculated. The mRNA and protein expressions were found to be increased in animals given anxiety as compared to the normal control. Whereas, the expressions were decreased in the animals treated with eugenol and its liposome-based nanocarriers in a dose dependent manner. However, the results were better in animals treated with nanocarriers as compared to the compound alone. It is concluded that the eugenol and its liposome-based nanocarriers exert anxiolytic activity by down-regulating GLO-1 protein expression in mice.

Resumo A forma mais comum de disfunção psicossocial é a ansiedade intimamente relacionada com a depressão, sem qualquer barreira de idade. Elas estão presentes em um estado combinado de tristeza, confusão, estresse, medo etc. O sistema de glioxalase contém uma enzima chamada glioxalase 1 (GLO1). É uma via metabólica que desintoxica alfa-oxo-aldeídos, particularmente metilglioxal (MG). O metilglioxal é produzido principalmente pela quebra dos intermediários glicolíticos, gliceraldeído-3-fosfatos e fosfato de diidroxiacetona. A expressão da glioxalase 1 também está relacionada ao comportamento de ansiedade. Um papel casual ou GLO1 no comportamento de ansiedade usando vetores virais para superexpressão no córtex cingulado anterior foi encontrado e descobriu-se que a superexpressão local de GLO1 aumentava o comportamento de ansiedade. O presente estudo trata do mecanismo molecular da atividade protetora do eugenol contra o transtorno ansiolítico. Um estudo pré-clínico em animais foi realizado em 42 camundongos BALB / c. Os animais foram submetidos ao estresse por meio do modelo de contenção convencional. A expressão de mRNA de GLO1 foi analisada por RT-PCR em tempo real. Além disso, a expressão da proteína GLO1 também foi examinada por imuno-histoquímica em todo o cérebro e a densidade média foi calculada. Verificou-se que as expressões de mRNA e proteínas estavam aumentadas em animais que receberam ansiedade em comparação com o controle normal. Considerando que as expressões foram diminuídas nos animais tratados com eugenol e seus nanocarreadores baseados em lipossomas de forma dependente da dose. No entanto, os resultados foram melhores em animais tratados com nanocarreadores em comparação com o composto sozinho. Conclui-se que o eugenol e seus nanocarreadores baseados em lipossomas exercem atividade ansiolítica por regulação negativa da expressão da proteína GLO1 em camundongos.

Eugenol and its liposome-based nano carrier reduce anxiety by inhibiting glyoxylase-1 expression in mice / Eugenol e seu nanocarreador à base de lipossoma reduzem a ansiedade ao inibir a expressão de glioxalase 1 em camundongos

Abstract The most common form of psycho-social dysfunction is anxiety with depression being related closely without any age bar. They are present with combined state of sadness, confusion, stress, fear etc. Glyoxalase system contains enzyme named glyoxalase 1 (GLO1).It is a metabolic pathway which detoxifies alpha-oxo-aldehydes, particularly methylglyoxal (MG). Methylglyoxal is mainly made by the breakdown of the glycolytic intermediates, glyceraldehyde-3-phosphates and dihydroxyacetone phosphate. Glyoxylase-1 expression is also related with anxiety behavior. A casual role or GLO-1 in anxiety behavior by using viral vectors for over expression in the anterior cingulate cortex was found and it was found that local GLO-1 over expression increased anxiety behavior. The present study deals with the molecular mechanism of protective activity of eugenol against anxiolytic disorder. A pre-clinical animal study was performed on 42 BALB/c mice. Animals were given stress through conventional restrain model. The mRNA expression of GLO-1 was analyzed by real time RT-PCR. Moreover, the GLO-1 protein expression was also examined by immunohistochemistry in whole brain and mean density was calculated. The mRNA and protein expressions were found to be increased in animals given anxiety as compared to the normal control. Whereas, the expressions were decreased in the animals treated with eugenol and its liposome-based nanocarriers in a dose dependent manner. However, the results were better in animals treated with nanocarriers as compared to the compound alone. It is concluded that the eugenol and its liposome-based nanocarriers exert anxiolytic activity by down-regulating GLO-1 protein expression in mice.

Resumo A forma mais comum de disfunção psicossocial é a ansiedade intimamente relacionada com a depressão, sem qualquer barreira de idade. Elas estão presentes em um estado combinado de tristeza, confusão, estresse, medo etc. O sistema de glioxalase contém uma enzima chamada glioxalase 1 (GLO1). É uma via metabólica que desintoxica alfa-oxo-aldeídos, particularmente metilglioxal (MG). O metilglioxal é produzido principalmente pela quebra dos intermediários glicolíticos, gliceraldeído-3-fosfatos e fosfato de diidroxiacetona. A expressão da glioxalase 1 também está relacionada ao comportamento de ansiedade. Um papel casual ou GLO1 no comportamento de ansiedade usando vetores virais para superexpressão no córtex cingulado anterior foi encontrado e descobriu-se que a superexpressão local de GLO1 aumentava o comportamento de ansiedade. O presente estudo trata do mecanismo molecular da atividade protetora do eugenol contra o transtorno ansiolítico. Um estudo pré-clínico em animais foi realizado em 42 camundongos BALB / c. Os animais foram submetidos ao estresse por meio do modelo de contenção convencional. A expressão de mRNA de GLO1 foi analisada por RT-PCR em tempo real. Além disso, a expressão da proteína GLO1 também foi examinada por imuno-histoquímica em todo o cérebro e a densidade média foi calculada. Verificou-se que as expressões de mRNA e proteínas estavam aumentadas em animais que receberam ansiedade em comparação com o controle normal. Considerando que as expressões foram diminuídas nos animais tratados com eugenol e seus nanocarreadores baseados em lipossomas de forma dependente da dose. No entanto, os resultados foram melhores em animais tratados com nanocarreadores em comparação com o composto sozinho. Conclui-se que o eugenol e seus nanocarreadores baseados em lipossomas exercem atividade ansiolítica por regulação negativa da expressão da proteína GLO1 em camundongos.

Machanism of methyleugenoc in protecting islet cells from hypoxia/reoxygenation injury / 中华器官移植杂志

Objective:To explore the protective effect of methyl eugenol (Me) on islet ischemia/reperfusion (I/R) injury and elucidate its underlying mechanism.Methods:The islets were isolated and purified from 6-8 week male BALB/c mice and divided into four groups of normal control (normal culture without any treatment), hypoxia/reoxygenation (H/R treatment), H/R+ dimethyl sulfoxide (DMSO dosing plus H/R treatment) and H/R+ Me (Me dosing plus H/R). Viability of islet cells in each group was detected by acridine orange (AO)/propidium iodide (PI) double stain.Function of islet cells (insulin secretion) was measured by enzyme-linked immunosorbent assay (ELISA). Murine islet β Min6 cells were selected for detecting the effect of Me on the proliferative activity of normal cultured and H/R treated islet cells under different concentration gradients by CCK8.Then Min6 cells were divided into four groups of normal, H/R, H/R+ DMSO and H/R+ Me.The definition of group was the same as that of primary murine islets.Flow cytometry and Hoechst 33342 nuclear stain were utilized for detecting cell apoptotic rate in each group.The protein expressions of p-JNK, p-p38, JNK, p38, Bcl-2 and Bax were detected by Western blot.And the data were processed by one-way ANOVA or t test.Results:The proportion of dead islet cells in H/R group was (29.47±2.65)% and it was significantly lower than that in normal group (7.63±1.53)%.And the inter-group differences were statistically significant ( P<0.001). The proportion of dead islet cells was (20.63±3.07)% in H/R+ Me group.It was higher than that in H/R group (29.47±2.65)% and in H/R+ DMSO group (30.13±1.50)% and inter-group difference was statistically significant ( P<0.05 & P<0.01). Under the stimulation of high glucose, the insulin secretion level of islet in H/R+ Me group was (1.76+ 0.08) mg/L, which was higher than that in H/R group and H/R+ DMSD group(1.24±0.14)mg/L and(1.27±0.05)mg/L, and the difference was statistically significant[(1.76±0.08) vs. (1.24±0.14) mg/L; (1.76±0.08) vs.(1.27±0.05) mg/L, P<0.01]. There was no significant effect on cell viability after Me dosing within a certain concentration range (0-40 μmol/L). After Me dosing (5 μmol/L), cell viability of H/R-treated Min6 cells was significantly higher than that without Me.And the difference was statistically significant[(1.19±0.03) vs.(1.00±0), P<0.01]. As compared with H/R and H/R+ DMSO groups, overall apoptotic rate declined in H/R+ Me group (Hoechst 33342 stain: 14.50%±1.05% vs. 23.30%±1.18%, 14.50%±1.05% vs. 22.77%±1.75%, P<0.001; Flow cytometry: 4.36%±0.54% vs. 21.44%±1.02%, 4.36%±0.54% vs. 21.68%±3.06%, P<0.01). The expressions of p-JNK and p-p38 were down-regulated (p-JNK: 0.77±0.06 vs. 1.03±0.05, 0.77±0.06 vs.0.93±0.04, P<0.001; p-p38: 0.80±0.05 vs. 1.01±0.08; 0.80±0.05 vs. 1.00±0.05, P<0.05) while Bcl-2/Bax ratio rose (1.62±0.13 vs. 0.72±0.10, 1.62±0.13 vs. 0.74±0.13, P<0.01). Conclusions:Me can improve the viability and function of islets and suppress the apoptosis of Min6 cells after H/R.The mechanism is correlated with JNK and p38 MAPK signaling pathways.

Anti-proliferating effect of Ocimum sanctum and Centella asiatica plant extract on growth of human glioblastoma cells: An in vitro study

Glioblastoma is aggressive brain tumour with poor prognosis with conventional chemotherapy, hence there is need to find alternative targets for developing newer treatment. Advent of new treatment methods involving medicinal plants have shown to reduced Cancer mortalities and prevents development of drug resistance for chemotherapy. Present study aimed at investigates the anti-proliferating activity of two promising medicinal plants, Ocimum sanctum and Centella asiatica. We studied the effect of their plant extract on U87MG Glioblastoma cells proliferation, survival effect and apoptosis. Cytotoxic activity was assessed, after the plant extract treatment on U87MG using MTT assay with dose of 1 mg/mL to 25mg/mL and apoptosis assess was done using Annexin V assay with the three dose (1.5 mg/mL, 2 mg/mL and 2.5 mg/mL). Survivin gene expression was studied using QRT-PCR (Rotar gene Q, Qiagene) has a marker of proliferation. Ocimum sanctum and Centella asiatica treatment of U87MG cells with dosage of 1.5 mg/mL, 2.0 mg/mL, 2.5 mg/mL showed increase in mean apoptotic cells 2.8 %, 4.9%, 10 % and 3.1%, 5.8% and 7.2%, respectively, compared to untreated U87MG cells. Survivin gene analysis of U87MG cells showed down-regulation in gene expression and differences was significant in comparison to untreated control group with both the plant extract, Centella asiatica showed more down-regulation (97% with 2.5 mg/mL) than Ocimum sanctum. Ocimum sanctum and Centella asiatica exhibited promising anti-proliferating activity and induces apoptosis by down regulation of survivin gene expression

ANTI-PARKINSON POTENTIAL OF OCCIMUM SANTUM THROUGH IN-SILICO DOCKING STUDY

Objective: Parkinson's disease is affecting millions of people worldwide. The prevalence of Parkinson's disease is 0.3% globally, rising to 1% in more than 60 years of age and 4% in more than 80 years of age and the figures are thought to be doubled by 2030. Thus, there is a great need to identify novel therapeutic strategies or candidate drug molecule which can rescue neuronal degeneration. The aim of the present study was to assess bioactive compounds found in Tulsi as potential antiparkinson activity using molecular docking and to provide scientific justification in term of its active ingredient to target protein for prevention and symptomatic treatment of diabetics. The active compounds of Ocimum sanctum is to reveal its pote Methods: ntiality by molecular docking analysis to find out its potent compound against parkinsonism which was done by Lipinski's rule in docking analysis. A wi Results: de range of docking score found during molecular docking analysis. Among the compounds Alpha-farnesene showed the highest negative value which is the best dock-score i.e., -6.2 followed by Cyclohexane-1,2,4- triethenyl (-5.9) followed by Benzene, 1, 2- dimethoxy-4-(1-propenyl) (-5.7) followed by Eugenol (-5.2). Conclusion: Alpha-farnesene and Cyclohexane-1,2,4- triethenyl are the best compounds for inhibiting of both, as it possessed best value in Molecular docking hence these are the potent antiparkinsonism agent.

Chemical composition and larvicidal activity of the essential oil of Pimenta dioica leaves / Composición química y actividad larvicida del aceite esencial de hojas de Pimenta dioica

In this study, we investigated the main constituent, the predominant class and biological activity of the essential oil extracted from the leaves of Pimenta dioica and the pattern of the major constituent against larvae in the third stage of Aedes aegypti. For this reason, we extracted the oil by hydrodistillation, identified its components by gas chromatography coupled with mass spectrometry (GC/MS) and calculated the lethal concentration (LC50) of the larvicidal activity using the Reed-Muench method. The results show that the oil consists mainly of eugenol, in which the phenylpropanoid class predominated and the lethal concentration, LC50, was 38.86 µg mL-1at a confidence level of 2.25 µg mL-1, while the eugenol standard presented LC5079.75 µg mL-1at a confidence level of 2.10 µg mL-1. Given the facts, we conclude that the oil is more active than the standard and that it has the potential to replace chemical larvicides.

En este estudio, investigamos el constituyente principal, la clase predominante y la actividad biológica del aceite esencial extraído de las hojas de Pimenta dioica y el patrón del constituyente principal contra las larvas en la tercera etapa de Aedes aegypti. Por este motivo, extrajimos el aceite por hidrodestilación, identificamos sus componentes mediante cromatografía de gases acoplada a espectrometría de masas (GC/MS) y calculamos la concentración letal (CL50) de la actividad larvicida mediante el método Reed-Muench. Los resultados muestran que el aceite está constituido principalmente por eugenol, en el que predominó la clase fenilpropanoide y la concentración letal, CL50, fue de 38,86 µg.mL-1 a un nivel de confianza de 2,25 µg.mL-1, mientras que el estándar de eugenol presentó CL50 79,75 µg.mL -1 a un nivel de confianza de 2,10 µg.mL-1. Dados los hechos, concluimos que el aceite es más activo que el estándar y que tiene el potencial de reemplazar los larvicidas químicos.

In vitro and in silico evaluation of the schistosomicidal activity of eugenol derivatives using biochemical, molecular, and morphological tools

Background Eugenol shows both antibacterial and antiparasitic activities, suggesting that it might be evaluated as an option for the treatment of praziquantel-resistant schistosome. Methods The in vitro activities of three eugenol derivatives (FB1, FB4 and FB9) on adult worms from Schistosoma mansoni were examined by fluorescence and scanning electron microscopy to analyze effects on the excretory system and integument damage, respectively. Biochemical tests with verapamil (a calcium channel antagonist) and ouabain (a Na+/K+-ATPase pump inhibitor) were used to characterize eugenol derivative interactions with calcium channels and the Na+/K+-ATPase, while in silico analysis identified potential Na+/K+-ATPase binding sites. Results The compounds showed effective doses (ED50) of 0.324 mM (FB1), 0.167 mM (FB4), and 0.340 mM (FB9). In addition, FB4 (0.322 mM), which showed the lowest ED50, ED90 and ED100 (p < 0.05), caused the most damage to the excretory system and integument, according to both fluorescence and scanning electron microscopy analysis. The death of adult worms was delayed by ouabain treatment plus FB1 (192 versus 72 hours) and FB9 (192 versus 168 hours), but the response to FB4 was the same in the presence or absence of ouabain. Besides, no changes were noted when all of the eugenol derivatives were combined with verapamil. Moreover, FB1 and FB9 inhibited Na+/K+-ATPase activity according to in silico analysis but FB4 did not show a time-dependent relationship and may act on targets other than the parasite Na+/K+-ATPase. Conclusion Eugenol derivatives, mainly FB4 when compared to FB1 and FB9, seem to act more effectively on the integument of adult S. mansoni worms.(AU)

Comparative study on in vivo and in vitro permeability of Huoxue Zhitong gel patch and microemulsion gel / 中国中药杂志

This study aims to establish a method for determination of paeonol(Pae), eugenol(Eug), and piperine(Pip) content in receptor liquid and research on the permeability and pharmacokinetics of Huoxue Zhitong gel patch and microemulsion gel. The Franz diffusion experiment was conducted to assess the percutaneous permeability, and the microdialysis method was employed to assess pharmacokinetics of Huoxue Zhitong gel patch and microemulsion gel. The content of Pae, Eug, and Pip in receptor liquid in vitro and in vivo was determined by HPLC and UPLC-MS. The Q_n and J_(ss) of Pae, Eug, and Pip in the gel patch were significantly higher than those in the microemulsion gel, indicating that the drug release was faster in the gel patch. The C_(max), AUC_(0-760), and MRT of Pae, Eug, and Pip in the gel patch were higher than those in the microemulsion gel, indicating that the gel patch can promote the penetration and prolong the skin residence of the drug. The results of this study provide reference for improving the dosage form of Huoxue Zhitong patch.

The hypoglycemic effect of eugenol on type 2 diabetic mice and regulation of signaling transduction pathway of glucose and lipid metabolism in liver / 中国药理学通报

Aim To study the effects of eugenol on hypoglycemic effect and hepatic glucose and lipid metabolism in type 2 diabetic mice, and to explore the possible mechanism. Methods The model of type 2 diabetes induced by long term high-fat diet was divided into four groups. The blood glucose and body weight of each group were measured once a week. After six weeks, the liver tissues of mice in each group were dissected and the liver mass and body mass of mice were weighed. Liver index, lipid metabolism and liver function were measured. Oral glucose tolerance test was performed. The levels of blood glucose, insulin, triglyceride, cholesterol, resistin, leptin, auxin, glucagon and plasminogen activator inhibitor-1 in serum were measured. He staining was used to observe the pathological changes of liver tissues. The expressions of SHP, pfoxo1, pCREB, PEPCK and G6Pase proteins in liver were detected by Western blot. Results Compared with HFD group, E40 group had lower body weight, smaller liver volume and healthy dark red. Compared with HFD group, E40 group decreased liver index, lipid metabolism and liver function. OGTT test showed that glucose tolerance was enhanced and the area under the curve was decreased in E40 group compared with HFD group. The levels of blood glucose, insulin, triglyceride, resistin, leptin, glucagon and plasminogen activator inhibitor-1 in E40 group were lower than those in HFD group. He staining showed that hepatocytes in HFD group were larger and accompanied with bullous steatosis than those in RD group. Hepatocyte steatosis and liver pathological state were significantly improved in E40 group. The results of Western blot showed that compared with HFD group, the expression of SHP, pfoxo1 and pCREB protein in E40 group was up-regulated, and the expression of PEPCK and G6Pase protein was down-regulated. Conclusions Eugenol can regulate the SHP/pFOXO1/PCREB/PEPCK/G6Pase signaling pathway, regulate glucose and lipid metabolism, inhibit insulin resistance, improve blood glucose level and glucose and lipid metabolism disorders in type 2 diabetes mellitus.

Fabrication, Physicochemical and Rheological Characterisation of a Drug-therapeutic Oils (Doxycycline Hyclate-Nigella sativa-Eugenol) Complex Emulsion Stabilised by Lecithin and Hydroxypropyl Methylcellulose Intended for Delivery Into Periodontal Pocket

@#Introduction: Earlier attempts to stabilise an emulsion, intended for chronic periodontitis treatment which composed of doxycycline hyclate (DH), Nigella sativa oil (NSO), eugenol and several combinations of surfactants failed. To solve the issue, we investigated the ability of lecithin alone and its combination with hydroxypropyl methylcellulose (HPMC) to stabilise the emulsion. Method: Compatibility between DH and other ingredients was first investigated using DSC and ATR-IR. The emulsion was characterised, firstly by preparing three phases: doxycycline/preservatives with or without HPMC (varying quantities), NSO/eugenol and lecithin/surfactants as aqueous, oil and emulsifier phases, respectively. The phases were added and emulsified sequentially at 7000 rpm (10 min) with an overhead stirrer and then at 3000 rpm (15min) using a high-shear mixer. DH assay was performed using validated HPLC method. Results: All ingredients were found to be compatible with doxycycline based on DSC, ATR-IR and supported by acceptable recovery (98.2±2.2 %) of DH from the emulsion. Stable emulsions were produced with particle size of 198.6±8.2 to 279.3±10.7 nm and zeta potential of -48.2±0.4 to -64.0±3.9 mV. The emulsions showed high viscosity (~200 Pa.s) at zero shear rate and exhibited shear-thinning flow upon increased in shear stress yielding viscosity of ~3 Pa.s at 100 s-1 indicating pseudoplastic behaviour suitable for pre-filled syringe packaging intended for delivery into periodontal pocket. Conclusion: Lecithin is an excellent emulsifier that can also impart pseudoplasticity for a complex emulsion constitute of drug and natural oils. This could pave the way for a more complex emulsion formulation fusing contemporary and therapeutic oils

Market survey of Caryophylli Flos specifications / 中国中药杂志

The present study aimed to regulate the market circulation of Caryophylli Flos and formulate standards for commodity specifications and grades of Caryophylli Flos. Market survey was carried out in four major medicinal material markets with 48 samples of Caryophylli Flos collected. The property, 100-seed weight, impurity percentage, moisture, and eugenol content in Caryophylli Flos of different specifications from different producing areas were determined and analyzed. The results showed that 27.1% of the samples surveyed on the markets did not meet the requirements of Chinese Pharmacopoeia(2020 edition). The 100-seed weight and the property are important factors for the classification of Caryophylli Flos specifications. There were significant differences in the property, 100-seed weight, impurity percentage, and eugenol content in Caryophylli Flos samples of different specifications from different producing areas, and also differences in the proportions of different specifications in Caryophylli Flos samples from different producing areas. The African-originated Xiaohong(medium grade) and Guangxi-originated Xiaohong(medium grade) accounted for 70% and 66.7% respectively, the Indonesian-originated Dahong(top grade) for 56.2%. In conclusion, there are many problems in the circulation of Caryophylli Flos at present, mainly including the loss of origin information, no standards for specifications, non-implementation of grade standards, excessive impurities, and no evidence for authenticity identification. According to the classification of Caryophylli Flos specifications in this study, the average eugenol content of Xiaohong is significantly higher than the Dahong by 4.74%.

Effect of methyl eugenol on hypoxia/reoxygenation injury of human renal tubular epithelial cells and its mechanism / 中国中药杂志

This study aimed to investigate the effect of methyl eugenol(ME) on hypoxia/reoxygenation(H/R)-induced injury of human renal tubular epithelial HK-2 cells and its mechanism. The viability of HK-2 cells cultured with different concentrations of ME and exposed to H/R was detected by cell counting kit-8(CCK-8) assay. The effect of ME on the morphology of HK-2 cells was observed under an inverted microscope. The content of intracellular reactive oxygen species in different groups was detected after 2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA) fluorescence staining. Cell apoptosis was determined by flow cytometry. Changes in mitochondrial membrane potential were monitored by JC-1 dye. The concentrations of nuclear factor erythroid 2 related factor 2(Nrf2), heme oxygenase-1(HO-1), and nicotinamide adenine dinucleotide phosphatase oxidase 4(Nox4) were measured by Western blot, followed by the assay of Nrf2 concentration changes in cytoplasm and nucleus by confocal fluorescence staining. The results showed that when the concentration of ME was 0-40 μmol·L~(-1), the activity of HK-2 cells was not affected. Compared with the model group, ME enhanced the activity of HK-2 cells and the cell morphology was normal. As revealed by further experiments, ME inhibited the release of reactive oxygen species and the decline in mitochondrial membrane potential of HK-2 cells after H/R injury, promoted Nrf2/HO-1 expression and Nrf2 translocation to the nucleus, and down-regulated the expression of Nox4, thereby significantly reducing apoptosis. This protective effect of ME could be reversed by the specific Nrf2 inhibitor ML385. These findings have preliminarily proved that ME effectively protected HK-2 cells against H/R injury, which might be related to its promotion of Nrf2/HO-1 signaling pathway and inhibition of Nox4. Such exploration on the possible mechanism of ME in the treatment of renal ischemia-reperfusion injury(IRI) and protection of organ function from the perspective of antioxidant stress has provided reference for related research on the treatment of acute kidney injury with traditional Chinese medicine.

Infiltration by Streptococcus mutans in provisional crowns fixed by two types of cementing agents ­ pilot double-blind randomized clinical trial / Infiltração por Streptococcus mutans em coroas dentárias provisórias fixadas por dois tipos de agentes cimentantes ­ estudo clínico randomizado duplo-cego piloto

Objective: When provisional acrylic crowns are used for a long time, they become more susceptible to marginal leakage by cariogenic bacteria. The objectives of this pilot clinical study were to compare cement based on zinc oxide-eugenol and calcium hydroxide by contamination with Streptococcus mutans, and calculate the sample size for the continuation of this study. Methods: Individuals receiving provisional crowns and following inclusion/exclusion criteria, were randomly distributed into 2 groups: zinc oxide-eugenol (n=8); calcium hydroxide (n=9). The temporary crowns were made by a blind researcher and cemented by another. Patients were also blinded by the cement used inside their crowns. After 2 months, a cement sample from the crowns' peripheral inner face was collected, placed in a tube containing 1 mL of sterile saline, serially diluted, plated on Mitis Salivarius Bacitracin agar, and incubated for 48 hours. Colony-forming units (CFU/mL) were counted. A statistical power analysis was performed to calculate sample size (1-ß=80%) and the Mann Whitney test to compare both cements (α=0.05). Results: Both cements were contaminated with S. mutans, with an average of 166.6 x 102 CFU/mL for calcium hydroxide and 435.3 x 102 CFU/mL for zinc oxide-eugenol, with no significant difference (p=0.311). The sample size calculated for this study was 36 per group. Conclusion: This pilot study suggests that there is important contamination inside provisional crowns used for two months, independent of the cement. The continuation of this study is needed, with a bigger sample size, to enable a comparison between the cements.

Objetivo: Quando coroas dentais provisórias são utilizadas por um longo período, elas se tornam susceptíveis à infiltração marginal por bactérias cariogênicas. O objetivo deste estudo clínico piloto foi comparar os cimentos a base de óxido de zinco e eugenol e hidróxido de cálcio pela contaminação com Streptococcus mutans e calcular o tamanho amostral para continuação deste estudo. Métodos: Indivíduos recebendo coroas provisórias e seguindo critérios de inclusão/exclusão, foram distribuídos aleatoriamente em dois grupos: óxido de zinco e eugenol (n=8); hidróxido de cálcio (n=9). As coroas provisórias foram feitas por um pesquisador cego e cimentadas por outro. Os pacientes também foram cegos quanto ao cimento utilizado dentro de suas coroas. Depois de 2 meses, amostras de cimentos foram coletadas da face interna periférica das coroas, colocadas em um tubo contendo 1 mL de solução salina estéril, diluídas de forma seriada, plaqueadas em ágar Mitis Salivarius Bacitracina e incubadas por 48 horas. Unidades formadoras de colônias (UFC/mL) foram contadas. Um teste de poder estatístico foi realizado para calcular o tamanho amostral (1-ß=80%) e o teste de Mann Whitney para comparar os dois cimentos (α=0.05). Resultados: Os dois cimentos foram contaminados com S. mutans, com uma média de 166.6 x 102 UFC/mL para o hidróxido de cálcio e 435.3 x 102 UFC/mL para o óxido de zinco e eugenol, com nenhuma diferença significativa (p=0.311). O cálculo amostral para este estudo foi 36 indivíduos por grupo. Conclusão: Este estudo piloto sugere que existe importante contaminação dentro de coroas provisórias utilizadas por 2 meses, independente do cimento. A continuação deste estudo é necessária, com maior tamanho amostral, para possibilitar a comparação entre os cimentos.

Methyl Eugenol, 1,8-Cineole and Nerolidol Rich Essential Oils with their Biological Activities from three Melaleuca Species Growing in Tarai Region of North India

Abstract The essential oils from the fresh leaves of three Melalecua spp. viz; Melaleuca leucadendron (L.) Melaleuca linariifolia Sm. and Melaleuca bracteata F. Muell. growing in Tarai region of North India were analyzed by a combination of gas chromatography/mass spectrometry. The analysis revealed the presence of several constituents of industrial and pharmacological importance. M. leucadendron essential oil was found to be dominated by E-nerolidol (85.7%) rich chemotype. 1,8-cineole (61.1%) along with significant presence of α-terpineol (12.3%), α-pinene (4.0%), β-myrcene (3.8%), and E-caryophyllene (1.7%) were identified in the essential oil from M. linariifolia Similarly M. bracteata was dominated by the presence of phenylpropanoids viz; methyl eugenol (74.8%) and methyl cinnamate (8.0%). The essential oils were studied for their in-vitro antioxidant, anti-inflammatory and antimicrobial potential. All the oils revealed potential antioxidant activity with maximum in M. bracteata essential oil. All the oils exhibited significant antibacterial activity against Bacillus megaterium, Staphylococcus aureus, Escherichia coli,Salmonella typhimurium and anti-fungal activity against phytopathogenic fungi Fusarium oxysporum, Sclerotinia sclerotiorum, Exserohilum turcicum and Curvularia lunata. The observations from present study suggest further cultivation of Melaleucas and its commercialization as industrial crops.

A influência do eugenol na microdureza de duas resinas compostas comerciais / The influence of eugenol on the microhardness of two commercial composite resins

Objetivo: avaliar, in vitro, a dureza superficial e profunda de dois tipos diferentes de resina composta, quandoem contato com o cimento de óxido de zinco e eugenol (IRM®). Métodos: foram selecionadas as resinasCharisma® e Vittra® e confeccionados 80 corpos de prova, sendo 40 para cada marca de compósito, queforam divididos em oito grupos (n=10) conforme o contato (imediato, 7 dias, 14 dias) ou não (grupo controle)com o eugenol. Colocou-se num pote dappen uma porção de IRM e um espécime de resina pronto, que,após a presa do material restaurador temporário e a remoção do exemplar colocado, gerou o molde pararealização das amostras em contato com o eugenol, sendo armazenado em água em temperatura ambiente,para os subgrupos 7 e 14 dias. Em sequência, foi inserido um único incremento de resina composta e polimerizadopor 40 segundos, gerando os espécimes que foram armazenados em local seco para serem submetidosao teste de microdureza Vickers sob uma carga de 300 gramas, com tempo de penetração de 10 segundos.Os dados obtidos foram analisados pelo teste ANOVA, a um nível de significância de p<0,05. Resultados:somente o grupo da resina Charisma® que teve contato imediato com eugenol não atingiu 80% de durezaprofunda em relação à superficial. Entretanto, para todos os grupos analisados, tanto a dureza superficialquanto a profunda aumentaram após o contato com o eugenol em relação ao grupo controle. Conclusão: oeugenol não influenciou negativamente a dureza das resinas compostas.(AU)

Objective: to evaluate, in vitro, the superficial and deep hardness of two different types of composite resin when in contact with zinc oxide eugenol cement (IRM). Method: Charisma® and Vittra® resins were selected and 80 specimens were made, 40 for each composite brand that were divided into eight groups (n = 10) according to contact (immediate, 7 days, 14 days) or no (control group) with eugenol. A portion of IRM and a ready-made resin specimen were placed in a dappen pot, which after setting the temporary restorative material and removing the placed specimen, generated the template for making the samples in contact with eugenol, being stored in water in room temperature for subgroups 7 and 14 days. In sequence, a single increment of composite was inserted and polymerized resin for 40 seconds, generating specimens that were stored in a dry place to be submitted to the Vickers microhardness test under a load of 300 grams, with a penetration time of 10 seconds. The data obtained were analyzed by the ANOVA test, at a significance level of p <0.05. Results: only the Charisma® resin group that had immediate contact with eugenol did not reach 80% deep hardness in relation to superficial hardness. However, for all groups analyzed, both superficial and deep hardness increased after contact with eugenol compared to the control group. Conclusion: eugenol did not negatively influence the hardness of composite resins.(AU)