Optimization of Antioxidant Activity of Compounds Generated during Ginseng Extract Fermentation Supplemented with Lactobacillus.

Ginseng holds high medicinal and cosmetic value, with stem and leaf extracts garnering attention for their abundant bioactive ingredients. Meanwhile, fermentation can enhance the effectiveness of cosmetics. The aim of this study was to optimize ginseng fermentation to produce functional cosmetics. Ginseng stem and leaf extracts were fermented with five different strains of lactic acid bacteria. Using 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl radical (·OH), and superoxide anion (O2·-) scavenging activities as indicators, the fermentation process was optimized via response surface methodology. Finally, validation of the antioxidant activity of the optimized fermentation broth was performed using human skin cells (HaCaT and BJ cells). Based on the antioxidant potency composite comprehensive index, Lactiplantibacillus plantarum 1.140 was selected, and the optimized parameters were a fermentation time of 35.50 h, an inoculum size of 2.45%, and a temperature of 28.20 °C. Optimized fermentation boosted antioxidant activity: DPPH scavenging activity increased by 25.00%, ·OH by 94.00%, and O2·- by 73.00%. Only the rare ginsenoside Rg5 showed a substantial rise in content among the 11 ginsenosides examined after fermentation. Furthermore, the flavonoid content and ·OH scavenging activity were significantly negatively correlated (r = -1.00, p < 0.05), while the Rh1 content and O2·- scavenging activity were significantly positively correlated (r = 0.998, p < 0.05). Both the 0.06% (v/v) and 0.25% (v/v) concentrations of the optimized broth significantly promoted cell proliferation, and notable protective effects against oxidative damage were observed in HaCaT cells when the broth was at 0.06%. Collectively, we demonstrated that ginseng fermentation extract effectively eliminates free radicals, preventing and repairing cellular oxidative damage. This study has identified new options for the use of fermented ginseng in functional cosmetics.

Untargeted Metabolomic Analysis and Chemometrics to Identify Potential Marker Compounds for the Chemical Differentiation of Panax ginseng, P. quinquefolius, P. notoginseng, P. japonicus, and P. japonicus var. major.

The Panax L. genus is well-known for many positive physiological effects on humans, with major species including P. ginseng, P. quinquefolius, P. notoginseng, P. japonicus, and P. japonicus var. major, the first three of which are globally popular. The combination of UPLC-QTOF-MS and chemometrics were developed to profile "identification markers" enabling their differentiation. The establishment of reliable biomarkers that embody the intrinsic metabolites differentiating species within the same genus is a key in the modernization of traditional Chinese medicine. In this work, the metabolomic differences among these five species were shown, which is critical to ensure their appropriate use. Consequently, 49 compounds were characterized, including 38 identified robust biomarkers, which were mainly composed of saponins and contained small amounts of amino acids and fatty acids. VIP (projection variable importance) was used to identify these five kinds of ginseng. In conclusion, by illustrating the similarities and differences between the five species of ginseng with the use of an integrated strategy of combining UPLC-QTOF-MS and multivariate analysis, we provided a more efficient and more intelligent manner for explaining how the species differ and how their secondary metabolites affect this difference. The most important biomarkers that distinguished the five species included Notoginsenoside-R1, Majonoside R1, Vinaginsenoside R14, Ginsenoside-Rf, and Ginsenoside-Rd.

[Quality evaluation of Commelina communis from different origins based on multicomponent content determination combined with chemometrics].

This research established a high-performance liquid chromatography(HPLC) method for simultaneous determination of isoorientin, orientin, vitexin, and isovitexin in Commelina communis to conduct content difference analysis and quality evaluation of 62 batches of C. communis from different origins. The HPLC content determination was performed on a Dikma Platisil ODS chromatographic column(4.6 mm×250 mm, 5 µm), with acetonitrile-0.1% formic acid(14∶86) as the mobile phase. The detection wavelength was set at 348 nm, the flow rate was 1.0 mL·min~(-1), and the column temperature was 35 ℃. The differences in origins and quality of 62 batches of C. communis were studied by chemometrics. The results showed that the determination of four components mani-fested a good linear relationship in the range of mass concentration(r>0.999 9), and the average recovery rate was 96.17%-103.0%. The relative standard deviations(RSDs) of precision, stability, and repeatability were all less than 2.0%. The content of four components from high to low was isoorientin>isovitexin>orientin>vitexin. Forty-seven batches of C. communis with clear origins were classified into six categories by chemometrics. C. communis from different origins had different qualities. Generally, C. communis from Western China, Central China, and South of China had superior qualities. The HPLC method established in this study is specific, simple, and efficient, which provides references for the comprehensive evaluation of the quality of C. communis. The chemometrics shows that the qualities of C. communis from different origins are largely different. Isoorientin can be used as an index to determine the content of C. communis, and its content limit should be set no less than 0.023%.

Vector analysis of astigmatic correction after single-step transepithelial photorefractive keratectomy and femtosecond-assisted laser in-situ keratomileusis for low to moderate myopic astigmatism.

Purpose: This study aimed to evaluate the outcomes of astigmatic correction by single-step transepithelial photorefractive keratectomy (TransPRK) and femtosecond-assisted laser in-situ keratomileusis (Femto-LASIK) surgeries. Methods: A total of 218 subjects received TransPRK or Femto-LASIK surgery for the treatment of myopia and astigmatism (-2.25 to -0.25 D). Refraction errors and uncorrected (UDVA) and corrected distance visual acuity (CDVA) were examined before and at 3 months after surgery. Astigmatism changes were assessed by vector analysis. Results: Preoperative parameters of the TransPRK group were similar to the Femto-LASIK group. UDVA and CDVA at 3 months were similar between both groups. Manifest refraction (MR) spherical equivalent in the TransPRK group (0 ± 0.20 D) was slightly lower compared with the Femto-LASIK group at 3 months (0.11 ± 0.25 D, P = 0.001). MR cylinder was -0.06 ± 0.19 D in the TransPRK group and -0.02 ± 0.15 D in the Femto-LASIK group at 3 months (P = 0.135). The index of success (IS) was 0.15 ± 0.36 in the TransPRK group and 0.06 ± 0.17 in the Femto-LASIK group (P = 0.125). The correction index (CI) was 1.03 ± 0.19 in the TransPRK group and 1.01 ± 0.11 in the Femto-LASIK group (P = 0.815). Conclusion: For low to moderate myopic astigmatism, TransPRK provided a comparable astigmatic treatment effect as Femto-LASIK. Myopic astigmatism was both slightly overcorrected after TransPRK and Femto-LASIK surgeries.

Phytochemical and pharmacological progress on Syringa oblata, a traditional Mongolian medicine.

Syringa oblata is a traditional Mongolian medicine mainly distributed in the Helan Mountains (the boundaries of Inner Mongolia and Ningxia, China) and the north of Yan Mountains (Aohan Qi, Inner Mongolia, China). It is clinically used to treat diseases caused by Heyi, such as heartache and heat pathogen in the heart. Phytochemical studies on S. oblata revealed the presence of iridoids, lignans, triterpenes, phenylpropanoids, phenylethanoids, and volatile components. Pharmacological investigations revealed a broad spectrum of bioactivities, such as antimicrobial, antioxidant, antiproliferative, and hepatoprotective effects. This article summarized the chemical components and pharmacological activities of S. oblata, providing a scientific rationale for its bioactive constituents, quality control, and utilization as an important medicine.

Changes in effective optical zone after small-incision lenticule extraction in high myopia.

PURPOSE: To evaluate the four measurement approaches on the determination of effective optical zone (EOZ) using Scheimpflug tomography after small-incision lenticule extraction surgery in eyes with high myopia. SETTING: Corneal refractive surgery conducted in an eye hospital in southern China. DESIGN: This is a retrospective cohort study. METHODS: In total, 74 subjects were recruited. EOZ was measured at 3 months postoperatively using vertex-based (EOZV), pupil-based (EOZP), 4 mm-ring-based total corneal refraction method (EOZ4) and tangential curvature difference map method (EOZD), and their consistencies were compared. EOZs and planned optical zone (POZ) were compared and analyzed with eccentricity, ablation degree (AD) and total corneal aberrations. RESULTS: At 3 months after surgery, the mean root mean square of ΔHOA, ΔComa, ΔTrefoil and ΔSA were 0.53 ± 0.27 µm, 0.36 ± 0.20 µm, 0.01 ± 0.84 µm and 0.16 ± 0.14 µm, respectively. EOZV, EOZP, EOZ4 and EOZD were 5.87 ± 0.44 mm, 5.85 ± 0.45 mm, 4.78 ± 0.40 mm and 5.29 ± 0.27 mm, respectively, which were significantly smaller than POZ 6.48 ± 0.16 mm. Bland-Altman plots showed a good consistency among the four EOZs. The difference between the EOZV and EOZP was 0.02 mm within the range of clinically acceptable difference. In addition, the eccentricity was positively correlated with ΔHOA, ΔComa and ΔSA. CONCLUSIONS: All 4 measurement approaches demonstrated the reduction of EOZs compared to POZ. The EOZV was the closest to POZ, followed by EOZP. The ΔEOZs showed no significant difference with eccentricity, AD and corneal aberrations.

Comparative Transcriptomics and Metabolites Analysis of Two Closely Related Euphorbia Species Reveal Environmental Adaptation Mechanism and Active Ingredients Difference.

Roots of Euphorbia fischeriana and Euphorbia ebracteolata are recorded as the source plant of traditional Chinese medicine "Langdu," containing active ingredients with anticancer and anti-AIDS activity. However, the two species have specific patterns in the graphic distribution. Compared with E. ehracteolata, E. fischeriana distributes in higher latitude and lower temperature areas and might have experienced cold stress adaptation. To reveal the molecular mechanism of environmental adaptation, RNA-seq was performed toward the roots, stems, and leaves of E. fischeriana and E. ehracteolata. A total of 6,830 pairs of putative orthologs between the two species were identified. Estimations of non-synonymous or synonymous substitution rate ratios for these orthologs indicated that 533 of the pairs may be under positive selection (Ka/Ks > 0.5). Functional enrichment analysis revealed that significant proportions of the orthologs were in the TCA cycle, fructose and mannose metabolism, starch and sucrose metabolism, fatty acid biosynthesis, and terpenoid biosynthesis providing insights into how the two closely related Euphorbia species adapted differentially to extreme environments. Consistent with the transcriptome, a higher content of soluble sugars and proline was obtained in E. fischeriana, reflecting the adaptation of plants to different environments. Additionally, 5 primary or secondary metabolites were screened as the biomarkers to distinguish the two species. Determination of 4 diterpenoids was established and performed, showing jolkinolide B as a representative component in E. fischeriana, whereas ingenol endemic to E. ebracteolate. To better study population genetics, EST-SSR markers were generated and tested in 9 species of Euphorbia. A total of 33 of the 68 pairs were screened out for producing clear fragments in at least four species, which will furthermore facilitate the studies on the genetic improvement and phylogenetics of this rapidly adapting taxon. In this study, transcriptome and metabolome analyses revealed the evolution of genes related to cold stress tolerance, biosynthesis of TCA cycle, soluble sugars, fatty acids, and amino acids, consistent with the molecular strategy that genotypes adapting to environment. The key active ingredients of the two species were quantitatively analyzed to reveal the difference in pharmacodynamic substance basis and molecular mechanism, providing insights into rational crude drug use.

Icariin inhibits RANKL-induced osteoclastogenesis in RAW264.7 cells via inhibition of reactive oxygen species production by reducing the expression of NOX1 and NOX4.

Icariin (ICA), isolated from Herba Epimedii, is a natural flavonoid glycoside that possesses antioxidant properties and inhibits osteoclastogenesis. However, the mechanism underlying osteoclastogenesis inhibition by ICA remains unclear. Here, we investigated the effects of ICA on receptor activator of nuclear factor kappa-Β ligand (RANKL)-induced osteoclastogenesis in RAW264.7 cells. ICA inhibited the expression of osteoclastogenesis-related genes in RAW264.7 cells induced by RANKL. ICA could inhibit osteoclastogenesis without inhibiting the viability of RAW264.7 cells. In addition, ICA inhibited reactive oxygen species production in RANKL-induced RAW264.7 cells. ICA reduced the expression of nuclear factor in activated T cells, cytoplasmic 1, and tartrate-resistant acid phosphatase, which are osteoclast-related molecules. Moreover, ICA decreased the expression of nicotinamide adenine dinucleotide phosphate oxidase (NOX), specifically NOX1 and NOX4, in RANKL-induced RAW264.7 cells. Our findings suggest that ICA can be used as a potential therapeutic agent for osteolytic diseases such as osteoporosis.

Quercetin Improves Cardiomyocyte Vulnerability to Hypoxia by Regulating SIRT1/TMBIM6-Related Mitophagy and Endoplasmic Reticulum Stress.

Cardiomyocyte apoptosis is an important pathological mechanism underlying cardiovascular diseases and is commonly caused by hypoxia. Moreover, hypoxic injury occurs not only in common cardiovascular diseases but also following various treatments of heart-related conditions. One of the major mechanisms underlying hypoxic injury is oxidative stress. Quercetin has been shown to exert antioxidant stress and vascular protective effects, making it a promising candidate for treating cardiovascular diseases. Therefore, we examined the protective effect of quercetin on human cardiomyocytes subjected to hypoxia-induced oxidative stress damage and its underlying mechanism. Human cardiomyocytes were subjected to hypoxia/reoxygenation (H/R) in vitro with or without quercetin pretreatment; thereafter, flow cytometry, Cell Counting Kit-8 assay, laser scanning confocal microscopy, quantitative PCR, western blotting, and enzyme-linked immunosorbent assay were performed to analyze the effects of quercetin on cardiomyocytes. We found that H/R induced reactive oxygen species overproduction and endoplasmic reticulum stress, as well as inhibited the function of the mitochondria/endoplasmic reticulum and mitophagy, eventually leading to apoptosis and decreasing the viability of human cardiomyocytes. Quercetin pretreatment inhibited H/R-mediated overproduction of reactive oxygen species and damage caused by oxidative stress, increased mitophagy, regulated mRNA and protein expression of transmembrane BAX inhibitor-1 motif-containing 6 (TMBIM6), regulated endoplasmic reticulum stress, and improved the vulnerability of human cardiomyocytes to H/R. Furthermore, transfection with short interfering RNA against silent information regulator protein 1 (SIRT1) counteracted the protective effects of quercetin on cardiomyocytes. Thus, quercetin was predicted to regulate mitophagy and endoplasmic reticulum stress through SIRT1/TMBIM6 and inhibit H/R-induced oxidative stress damage. These findings may be useful for developing treatments for hypoxic injury-induced cardiovascular diseases and further highlight the potential of quercetin for regulating mitochondrial quality control and endoplasmic reticulum function.

Elucidation of metabolite isomers of Leonurus japonicus and Leonurus cardiaca using discriminating metabolite isomerism strategy based on ultra-high performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry.

The isomer structural discrimination is a significant challenge in metabolome analysis based on ultrahigh performance liquid chromatography tandem high-resolution mass spectrometry (UHPLC-HRMS). In this study, a new discriminating metabolite isomerism strategy is proposed to elucidate the metabolome, especially the isomers, of Leonurus japonicus and Leonurus cardiaca. This strategy consists of three steps. First, the metabolite biosynthesis pathways are constructed based on a home-built compound database to rapidly profile the compounds of interest using the multiple diagnostic product ions (DPIs) screening analysis and binary comparison based on SUMPRODUCT function. Second, the fragmentation patterns (e.g. the high-resolution DPIs, DPI ratios) and chromatographic elution order are defined based on scattered reference chromatographic and mass spectrometry data, calculated lipophilicity parameters, molecular hydrogen bond analysis, and chemical reference standards. Finally, all discovered isomerisms are mapped with the defined applicable rules and the isomers are identified conveniently. Using this strategy, a total of 257 compounds were tentatively characterized, including 212 potential novel compounds and 67 pairs of cis-, trans-, and positional isomers of flavonoids, phenylethanoid glycosides, glucaric acids, novel quinic acids, and esters of fatty acids. Moreover, 56 characteristic markers were identified to discriminate these two herbal medicines. This strategy may significantly improve the efficiency and reliability of identifying isomers found in metabolite biosynthesis pathways.

Biogenic Synthesis and Spatial Distribution of Endogenous Phytohormones and Ginsenosides Provide Insights on Their Intrinsic Relevance in Panax ginseng.

Ginseng, the root and rhizome of Panax ginseng C. A. Mey., is a well-known and valuable traditional Chinese medicine. The pharmacological activities of ginseng are mainly attributed to the presence of ginsenosides, which are considered to be critical biomarkers for evaluating ginseng quality. The biosynthesis of triterpenes, which serve defensive functions in plants, is regulated by endogenous phytohormones that play key roles in growth and defense of plant populations. However, the role of major hormones that are closely related to secondary metabolism pathways in P. ginseng is poorly understood. To gain insight into their potential correlation, we performed a spatial synthesis analysis and studied the distribution of endogenous phytohormones and ginsenosides in different tissue regions of the entire P. ginseng plant. Gibberellins are growth hormones that accumulate in the fiber root. In contrast, abscisic acid (ABA), salicylic acid (SA), and jasmonic acid (JA), which are considered stress hormones, were predominantly found in the leaf and leaf peduncle. We observed a tissue-specific distribution of phytohormones consistent with the expression of genes involved in hormone biosynthesis that influenced ginsenoside synthesis and distribution. The aim of this study was to investigate the role of different endogenous phytohormones on triterpene metabolites in ginseng innate immunity.

[Advances in research on relationships among Lycium species and origin of cultivated Lycium in China].

The Lycium genus consists of consist of 7 species and 3 variety which are main distributed in Northwest region in China, the cultivated Goji berry appeared about 1 000 years ago. The phylogeny of the wild Goji berry and the domestication of cultivated Goji berry are important scientific and practical value due to the medicinal and economic value. In this paper, a new advances achieved in studies on the phylogeny of wild Goji berry is summarized. The origin of cultivated Goji berry includes when, where, how and ancestral specie were reviewed. The current situation of cultivation and existing problems were summarized. We considered that through mutations and artificial selections, wild ancestors were domesticated to current cultivar groups. To find direct evidence for the origin of cultivated Goji berry, we must rely on genetic analysis in addition to morphological characters. To make full use of the Goji berry germplasm resources, the investigation and mining on wild Goji berry resource should be strengthened in the future.

[Characterization of tissue expression of ginsenoside biosynthetic gene expression in Panax ginseng].

The study is aimed to characterize the tissue expression of 10 key ginsenoside biosynthetic genes using bioinformatics method and real-time quantitative PCR. Heatmap and cluster analysis of 10 ginsenoside biosynthetic genes were performed in four-year-old Jilin ginseng. Using real-time quantitative PCR, the expression correlation of 10 key genes involved in ginsenoside biosynthesis was analyzed in different organs of four-year-old Jilin ginseng including, tissue culture seedling and adventitious root. Pearson correlation was used to analyze the relation between those 10 key genes involved in ginsenoside biosynthesis. The results showed that ß-AS and CYP716A52v2 were expressed highly in root of Jilin ginseng and ginseng culture seedling, which was consistent with Ro distribution. In addition, CYP716A53v2 and CYP716A47 which involved in dammarane type ginsenoside biosynthesis were positively correlated, which revealed that the difference of ginsenoside distribution was caused by transport system.

[Situation analysis and standard formulation of pesticide residues in traditional Chinese medicines].

Chinese Pharmacopoeia provides nine pesticide Maximum Residual Limits(MRLs) of traditional Chinese medicines(TCMs), The number of pesticides used in production are far more than those listed in pharmacopoeia. The lack of the standards make it's hard to reflect the real situation of pesticide residues in TCMs correctly. The paper is aimed to analyze the data of pesticide residues in TCMs from 7 089 items in 140 reports, and judging the exceedance rate of pesticides in TCMs using the MRLs of European pharmacopoeia，which is widely accepted in many countries. The results show that：①Pesticide residues in 18 kinds of TCMs are higher than MRLs，while in 137 kinds are below MRLs, such as Atractylodis Macrocephalae Rhizoma, Menthae Haplocalycis Herba and Fritillariae Thunbergii Bulbus. The average exceedance rate of all TCMs is 1.72%. The average exceedance rates of organochlorine, organophosphorus and pyrethroid are 2.26%, 1.51%, 0.37%，respectively. ②The average exceedance rate of pesticides is 2.00%, and the exceedance rate is more than 5%, accounting for 8.33%, the exceedance rate is between 1%-5%, accounting for 18.75%. the exceedance rate is between 0%-1%, accounting for 18.75%. The remaining 29 kinds of pesticides were not exceeded, accounting for 60.42%.Some reports like Greenpeace's organization exaggerated the pesticide residues in TCMs.But the pesticide residue question is still worthy of attention, so we proposed to amend the Chinese Pharmacopoeia pesticide residues standards, to increase the pesticide species of traditional Chinese medicine in production on the basis of retaining the existing types of pesticide residues, to strengthen the system research of pesticide residues in TCMs, providing a basis for making standard and promoting import and export trade in TCMs.

Study on dissolution and absorption of four dosage forms of isosorbide mononitrate: level A in vitro-in vivo correlation.

The objective of the present study was to develop a novel in vitro system to simulate the process of dissolution and permeation of oral solid dosage forms in vivo, and to establish a correlation between in vitro permeation and in vivo absorption that could predict the bioavailability (BA) and bioequivalence (BE) of congeneric products. The in vitro dissolution and absorption kinetics of four dosage forms of isosorbide mononitrate (ISMN) were evaluated by the USP basket/paddle system and drug dissolution/absorption simulating system (DDASS). The corresponding pharmacokinetic study was performed in beagle dogs. A comparative study was carried out between the classical and the novel method to estimate the effectiveness of the modified DDASS in simulating the course of dissolution and absorption in vivo. Indeed, the correlation coefficients of in vitro dissolution and in vivo absorption obtained from DDASS and dogs were higher. Moreover, a higher level A in vitro-in vivo correlation (IVIVC) between DDASS permeation and dog absorption was established, with correlation coefficients of 0.9968, 0.9872, 0.9921, and 0.9728. The DDASS method was more accurate at modeling the process of dissolution and absorption in vivo for both immediate-release (IR) and sustained-release (SR) dosage forms of ISMN.