Determination of Phenolic Acids Using Ultra-High-Performance Liquid Chromatography Coupled with Triple Quadrupole (UHPLC-QqQ) in Fruiting Bodies of Sanghuangporus baumii (Pilát) L.W. Zhou and Y.C. Dai.

Sanghuangporus, a medicinal mushroom, has gained significant attention due to its beneficial properties. Phenolic acids are among the major bioactive compounds in Sanghuangporus, known for their antioxidant and anti-inflammatory activities. To precisely quantify the phenolic acid content, we developed a method utilizing ultra-high-performance liquid chromatography with triple quadrupole (UHPLC-QqQ). This study optimized the UHPLC-QqQ conditions to simultaneously separate and detect eight phenolic acids in Sanghuangporus baumii (Pilát) L.W. Zhou and Y.C. Dai, including chlorogenic acid, p-coumaric acid, caffeic acid, cryptochlorogenic acid, protocatechuic acid, ferulic acid, sinapic acid, and syringic acid. The separation process utilized a ZORBAX Eclipse Plus C18 column using 0.01% formic acid and 2 mmol/L ammonium formate in water as the aqueous phase and methanol containing 0.01% formic acid and 2 mmol/L ammonium formate as the organic phase. Calibration curves were constructed using standard solutions to quantitatively determine the phenolic acid content. The results showed significant variation in phenolic acid content among S. baumii fruiting bodies, with Protocatechuic acid, p-coumaric acid, and caffeic acid being the most abundant. This method is valuable for quantifying phenolic acid compounds under different cultivation conditions. It provides excellent sensitivity, selectivity, and reproducibility for the quantification of phenolic acids in Sanghuangporus, contributing to a better understanding of its chemical composition and potential health benefits. This approach represents a novel technical means for the simultaneous analysis of compound phenolic acids in Sanghuangporus fruiting bodies.

Effects of Cultivation Methods on the Nutritional Content, Active Component Content, and Antioxidant Activity of Fruiting Bodies of Sanghuangporus baumii (Agaricomycetes).

To provide a scientific reference for improving the sawdust cultivation of Sanghuangporus baumii, comparative studies were conducted on the contents of nutritional components and active components and the antioxidant activity of the fruiting bodies of S. baumii cultivated with sawdust and cut logs. The results indicate that, first, cultivation methods had little effect on the contents of crude fat and the measured 16 kinds of amino acids [including total essential amino acids (EAA), total nonessential amino acids (NEAA), EAA/NEAA, and EAA/total amino acid (TAA)], but had a great influence on the contents of crude protein, crude fiber and TAA. These results suggest that the nutritional content under sawdust cultivation was significantly higher than that under cut-log cultivation. Second, the cultivation methods had little effect on the content of triterpenoids but had a great effect on the contents of polysaccharides, total flavonoids and total phenols, which showed that cut-log cultivation was significantly higher than sawdust cultivation. Third, the cultivation methods had a great effect on the antioxidant activities (ABTS and FRAP), which showed that cut-log cultivation was significantly higher than sawdust cultivation. The contents of polysaccharides, total flavonoids, and total phenols and the ABTS and FRAP activities using sawdust cultivation were lower than those using cut-log cultivation, which may be related to the mushroom strains, cultivation medium formula and cultivation technology. The results provide a solid basis for the improvement and promotion of new cultivation technologies for S. baumii.

Quantitative Analysis of Flavonoids in Fruiting Bodies of Sanghuangporus Using Ultra-High-Performance Liquid Chromatography Coupled with Triple Quadrupole Mass Spectrometry.

A rapid, precise, and dependable method for quantifying flavonoids in the fruiting bodies of Sanghuangporus was established using ultra-high-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC-QQQ-MS/MS). Separation was achieved using a ZORBAX Eclipse Plus C18 column (1.8 µm, 3.0 mm × 100 mm) with a 15 min gradient of a mobile phase consisting of 0.01% aqueous formic acid and 2 mm/L ammonium formate (mobile phase A), and 0.01% formic acid and 2 mm/L ammonium formate in methanol (mobile phase B). A mass spectrometry analysis was performed using the multiple reaction monitoring (MRM) mode with an electrospray ion source. This method enabled the simultaneous detection of 10 flavonoids (sakuranetin, quercitrin, myricitrin, kaempferol, luteolin, rutin, hyperoside, kaempferol-3-O-rutinoside, catechin, and catechin gallate) in the fruiting bodies of Sanghuangporus. Additionally, we applied this method to analyze the flavonoid content in fruiting bodies of various Sanghuangporus species. The results revealed substantial variations in flavonoid content, up to a 100-fold difference, among different species, with myricitrin, hyperoside, and rutin identified as the most abundant flavonoids. This protocol serves as a valuable tool for quantifying flavonoid compounds in different Sanghuangporus species or under diverse cultivation conditions, particularly for identifying species with high levels of specific flavonoid compounds.

Childhood parental companionship, self-esteem and prosocial behavior in college students: A correlation analysis.

BACKGROUND: We aimed to evaluate the correlation of Childhood parental companionship, self-esteem and prosocial behavior in college students. METHODS: We conducted a survey to assess the childhood parental companionship, self-esteem and prosocial behavior in our college from November 1, 2021 to December 15, 2021. The parental companionship status questionnaire, Self-Esteem Scale (SES) and prosocial behavior questionnaire were used for survey. Pearson linear correlation analysis was used for evaluating the correlation of childhood parental companionship, self-esteem and prosocial behavior in college students. The Bootstrap method was used to test the potentially mediating effect. RESULTS: A total of 2186 college students were included. The average total companionship score was (60.52 ± 5.17), the average self-esteem scale score was (27.15 ± 8.56), the prosocial behavior questionnaire score was (61.19 ± 15.04). Pearson correlation analysis indicated that childhood parental companionship was positively correlated with self-esteem (r = 0.679) and prosocial behavior(r = 0.679) in included college students (all P < 0.05). Self-esteem had mediating effect on parental companionship and prosocial behavior of included college students, its mediating effect was -0.445, accounting for 77.92 % of the total effect. CONCLUSIONS: Childhood parental companionship is positively correlated with self-esteem and prosocial behavior, and self-esteem play a mediating role in the parental companionship and prosocial behavior of college students.

Wenxin Granule Ameliorates Hypoxia/Reoxygenation-Induced Oxidative Stress in Mitochondria via the PKC-δ/NOX2/ROS Pathway in H9c2 Cells.

Myocardial infarction and following reperfusion therapy-induced myocardial ischemia/reperfusion (I/R) injury have been recognized as an important subject of cardiovascular disease with high mortality. As the antiarrhythmic agent, Wenxin Granule (WXG) is widely used to arrhythmia and heart failure. In our pilot study, we found the antioxidative potential of WXG in the treatment of myocardial I/R. This study is aimed at investigating whether WXG could treat cardiomyocyte hypoxia/reoxygenation (H/R) injury by inhibiting oxidative stress in mitochondria. The H9c2 cardiomyocyte cell line was subject to H/R stimuli to mimic I/R injury in vitro. WXG was added to the culture medium 24 h before H/R exposing as pretreatment. Protein kinase C-δ (PKC-δ) inhibitor rottlerin or PKC-δ lentivirus vectors were conducted on H9c2 cells to downregulate or overexpress PKC-δ protein. Then, the cell viability, oxidative stress levels, intracellular and mitochondrial ROS levels, mitochondrial function, and apoptosis index were analyzed. In addition, PKC-δ protein expression in each group was verified by western blot analysis. Compared with the control group, the PKC-δ protein level was significantly increased in the H/R group, which was remarkably improved by WXG or rottlerin. PKC-δ lentivirus vector-mediated PKC-δ overexpression was not reduced by WXG. WXG significantly improved H/R-induced cell injury, lower levels of SOD and GSH/GSSG ratio, higher levels of MDA, intracellular and mitochondrial ROS content, mitochondrial membrane potential and ATP loss, mitochondrial permeability transition pore opening, NOX2 activation, cytochrome C release, Bax/Bcl-2 ratio and cleaved caspase-3 increasing, and cell apoptosis. Similar findings were obtained from rottlerin treatment. However, the protective effects of WXG were abolished by PKC-δ overexpression, indicating that PKC-δ was a potential target of WXG treatment. Our findings demonstrated a novel mechanism by which WXG attenuated oxidative stress and mitochondrial dysfunction of H9c2 cells induced by H/R stimulation via inhibitory regulation of PKC-δ/NOX2/ROS signaling.

[Effects of nitrogen level on growth of Tetrastigma hemsleyanum and phytochemical content and antioxidant activity in stems and leaves].

As a rare endangered medical plant that newly cultivated,little experimental information is available for growth and metabolites of Tetrastigma hemsleyanum in response to nitrogen( N). The effects of different levels of N on growth of T. hemsleyanum and the content of phytochemicals( polysaccharide,total flavonoids and phenolics) and antioxidant activity( ABTS and FRAP) in stems and leaves were investigated in this study. A certain amount of N had positive effects on most of biological traits,and excessive dose of N went against growth of T. hemsleyanum. With N levels decreased,the polysaccharide content in stems and leaves had no significant change,while the total flavonoid and phenolic content,and antioxidant activities increased steadily. Antioxidant activities and total flavonoid and phenolic content had significant positive correlation. Excessive N fertilizer should be avoided by cultivation.

Association of various fruiting body macromorphological traits with spore yield in Ganoderma Lingzhi (higher Basidiomycetes), a new medicinal mushroom from China.

To study the correlations and relationship between spore yield and various macromorphological traits of Ganoderma lingzhi, a field experiment was conducted in a randomized block design with 3 replications. Ten macromorphological traits, including pileus diameter, pileus crust thickness, context thickness, tube thickness, pileus thickness, stipe length, stipe diameter, stipe weight, pileus weight, and spore yield, were recorded for all of the tested strains. There was significant variation among the strains for all of the traits studied. The results indicated that the highest variation was observed in spore yield and pileus weight. Correlation studies revealed that among 9 macromorphological traits, only the pileus weight of the fruiting body was significantly positively correlated with spore yield (r2 = 0.674*). Pileus diameter showed significant positive association with pileus weight of fruiting body (r2 = 0.838*). Stepwise multiple regression analysis showed that pileus weight and spore yield had a linear relationship (spore yield = -21.95 + 1.51 * pileus weight). The coefficient of determination of stepwise regression analysis (r2 = 0.4543) revealed 45.43% variation in the spore yield because of its relationship with pileus weight. Regression coefficient (b = 1.51) showed that a unit (1 g) increase in the pileus weight of the fruiting body resulted in a proportional increase of 1.51 g in spore yield. The derived information would be very useful when selecting potentially breeding strains for future G. lingzhi improvement programs.

Optimization of fermentation process for the production of intracellular polysaccharide from Paecilomyces cicadae and the immuno-stimulating activity of intracellular polysaccharide.

Optimization of fermentation process for the production of intracellular polysaccharide (IPS) from the fungus Paecilomyces cicadae and the immuno-stimulating activity of IPS were carried out. The quantitative effects of initial pH, fermentation temperature and time on the yield of IPS content produced by P. cicadae in submerged fermentation were investigated separately using response surface methodology (RSM). The three factors chosen for the present investigation were based on the results of a previous Plackett-Burman (PB) design. The experimental data obtained were fitted to a second-order polynomial equation using multiple regression analysis and also analyzed by appropriate statistical methods. RSM analysis showed good correspondence between experimental and predicted values. It was found that three parameters represented significant effect. Probability value (p < 0.0001) demonstrated a very high significance for the regression model. By solving the regression equation and analyzing the response surface contour plots, the optimal process parameters were determined, i.e. fermentation temperature 24.53 °C, initial pH 7.46 and fermentation time 73.9 h. The maximum predicted yield of IPS was 356.02 µg/ml under the optimal conditions. Meanwhile, IPS from P. cicadae was found to have direct immuno-stimulating activity in vitro on murine macrophage RAW264.7 proliferative response and to stimulate nitric oxide generation in a dose-dependent manner.

SCAR makers and multiplex PCR-based rapid molecular typing of Lentinula edodes strains.

Lentinula edodes is the second most important cultivated mushroom worldwide, the most commercial strains have been identified only through traditional phenotypic analysis. In this study, a simple rapid PCR-based molecular method was developed for distinguishing commercial strains of L. edodes by developing specific sequence characterized amplified region (SCAR) markers and establishing multiplex PCR assays with the SCAR primers. Derived from the randomly amplified polymorphic DNA (RAPD) and sequence-related amplified polymorphism (SRAP) techniques, 10 informative SCAR markers were generated from 10 polymorphic RAPD and SRAP bands. The differences in SCAR phenotypes among different strains made these SCAR markers potentially useful to characterize 6 strains and identify them from other studied strains. Moreover, different SCAR phenotypes also made the other 17 studied strains to be divided into four distinguishable groups. The multiplex PCR assays were further established for the joint use of some SCAR markers efficiently. Compared with some identification methods reported previously, the special feature of this new molecular method is technically rapid and convenient in the practical use and suitable for analyzing large numbers of samples. Thus, the simple rapid PCR-based molecular method can be used as a helpful assistant tool for the lentinula industry. To our knowledge, this study is the first to describe a development of a new SCAR maker-based multiplex PCR assay for rapid molecular typing of edible mushroom.

New available SCAR markers: potentially useful in distinguishing a commercial strain of the superior type from other strains of Lentinula edodes in China.

At present, more than 100 strains of Lentinula edodes are cultivated on a commercial scale in China. A simple, reliable, and effective method to distinguish some commercial strains of the superior type from other commercial strains is very important for the Lentinula industry. In this study, 23 commercial strains of L. edodes cultivated widely in China at present were collected and analyzed with randomly amplified polymorphic DNA (RAPD) technique. Three informative dominant sequence characterized amplified region (SCAR) markers were developed by designing three pairs of specific SCAR primers from three sequenced differential RAPD bands, respectively. Based on the three SCAR markers, three different multiplex polymerase chain reaction (PCR) phenotypes were detected among the 23 studied commercial strains and in which a multilocus phenotype characterizing a commercial strain Cr02 of the superior type could potentially be used to distinguish this strain from the other 22 studied commercial strains. To our knowledge, this study is the first to describe the development of a multiplex PCR technique based on SCAR markers for detecting the molecular phenotypes among commercial strains of L. edodes in China.