Qualitative and quantitative analysis of major components of Renshen-Yangrong Pill by UPLC-LTQ/Orbitrap/MS and UPLC-MS/MS.

Renshen-Yangrong Pill (RYP) is a classical traditional Chinese medicine (TCM) preparation for the treatment of asthenic symptoms, while its multiple herbal compositions bring a wide variety of unclear chemical components which seriously hinder the effective quality control and clinical practice. The present study aimed to investigate the overall chemical profile of RYP by UPLC-LTQ/Orbitrap/MS, and further obtain the quantitative distributions of representing components in the preparations. A total of 132 components in RYP including flavonoids, triterpenoid saponins, phenylpropanoids, and monoterpenoid glycosides were identified or tentatively characterized by authentic compounds or accurate masses and fragmentation, in which 52 characteristic components were selected for further quantitation by UPLC-MS/MS. The assay was validated in terms of linearity, precision, repeatability, recovery and successfully applied for the quality control of 40 batches of RYP. Hesperidin and paeoniflorin were revealed as the most abundant constituents in RYP, and the samples of different origins and dosage forms were clearly classified based on hierarchical cluster analysis. This study provided a deep insight into the chemical profiling of RYP, as well as a new approach for determining the marker compounds, which laid a valuable foundation for further investigation of potential effective components and comprehensive quality control of RYP and related preparations.

Effect of total parathyroidectomy in patients with secondary hyperparathyroidism: a retrospective study.

PURPOSE: To investigate the therapeutic efficacy, feasibility, and safety of total parathyroidectomy (tPTX) in the treatment of secondary hyperparathyroidism (SHPT). METHODS: The clinical data of 34 SHPT patients admitted to the Department of Nephrology, Yuxi People's Hospital, from January 2018 to January 2021 who had received tPTX, were retrospectively analyzed. The indications for tPTX were severe SHPT that did not respond to medical treatment and was ineligible for kidney transplantation. tPTX without autotransplantation was adopted to compare the level of symptom relief and changes in serum intact parathyroid hormone (iPTH), blood calcium, and blood phosphorus pre- and postoperatively. RESULTS: In 34 patients, 142 parathyroid glands were removed, including 21 ectopic parathyroid glands (14.78%). Six patients (17.64%, 6/34) had supernumerary parathyroid glands. At 6 h postoperatively, arthralgia and bone pain were significantly reduced to almost zero in 94.12% (32/34) of patients. At 24 h postoperatively, relief of bone pain and improvement of limb movement were observed in 100% (34/34) of patients, and pruritus almost disappeared in 86.36% (19/22) of patients. There were significant differences in iPTH (χ2 = 134.93, P < 0.05), calcium (χ2 = 23.02, P < 0.05), and phosphorus (χ2 = 102.11, P < 0.05) levels preoperatively and 40 min, 24 h, 1 week, half a year, and last available (> 1 year) postoperatively. The patients were followed up for 15-47 months (median 33 months). Hypoparathyroidism was observed in three patients, who underwent neck dissection or partial thymotomy concurrently for different reasons. No intractable hypocalcemia or adynamic bone disease occurred during the follow-up period. CONCLUSION: In SHPT patients who were ineligible for renal transplantation, tPTX was effective, safe, and reliable, with a low recurrence rate. However, when tPTX was performed alone without autologous transplantation, bilateral neck exploration was sufficient, and central neck dissection and thymic resection were inadvisable.

Development and evaluation of a chewing gum containing antimicrobial peptide GH12 for caries prevention.

The purpose of this study was to develop a chewing gum containing a novel antimicrobial peptide GH12 and evaluate its biocompatibility, antimicrobial activity, and caries-preventive effects in vivo and in vitro. GH12 chewing gum was developed using a conventional method and its extracts were prepared in artificial saliva. GH12 concentration in the extracts was determined by high-performance liquid chromatography; extracts were used for growth curve assay, time-kill assay, crystal violet staining assay, scanning electron microscopy, and Cell Counting Kit-8 assay. A rat caries model was established, and molars were treated topically with extracts for 5 weeks. Weight gain monitoring, hematoxylin-eosin staining, micro-computed tomography, and Keyes scoring were conducted. Significant inhibition of Streptococcus mutans growth and biofilm formation was observed. Extracts displayed low cytotoxicity against human gingival epithelial cells. No significant differences in weight gain or signs of harm to the mucosal tissues in any of the rats were observed. Keyes scores of caries lesions in the GH12 chewing gum group were lower than those of the negative control group. It was concluded that GH12 chewing gum showed good biocompatibility, antimicrobial activity, and caries-preventive effects, exhibiting great potential to prevent dental caries as an adjuvant to regular oral hygiene.

YY1 Promotes Telomerase Activity and Laryngeal Squamous Cell Carcinoma Progression Through Impairment of GAS5-Mediated p53 Stability.

Yin Yang 1 (YY1) is a key transcription factor that exerts functional roles in the cell biological process of various cancers. The current study aimed to elucidate the role and mechanism of YY1 in laryngeal squamous cell carcinoma (LSCC). YY1 mRNA and protein expression in human LSCC cell lines was detected by RT-qPCR and Western blot analysis. An interaction of YY1, GAS5, and p53 protein stability was predicted and confirmed by bioinformatics, ChIP, Co-IP, RIP, and FISH assays. Following loss- and gain-function assays, LSCC cell proliferation, colony formation, cell cycle, telomere length and telomerase activity were evaluated by CCK-8 assay, colony formation assay, flow cytometry, and PCR-ELISA, respectively. Nude mice were xenografted with the tumor in vivo. LSCC cell lines presented with upregulated expression of YY1, downregulated GAS5 expression, and decreased p53 stability. YY1 inhibited the expression of GAS5, which in turn recruited p300 and bound to p53, thus stabilizing it. Moreover, YY1 could directly interact with p300 and suppressp53 stability, leading to enhancement of cell proliferation, telomere length and telomerase activity in vitro along with tumor growth in vivo. Collectively, YY1 can stimulate proliferation and telomerase activity of LSCC cells through suppression of GAS5-dependent p53 stabilization or by decreasing p53 stability via a direct interaction with p300, suggesting that YY1 presents a therapeutic target as a potential oncogene in LSCC development and progression.

In Vivo and In Vitro Antioxidant Activities of Methanol Extracts from Olive Leaves on Caenorhabditis elegans.

The aim of this study was to evaluate the antioxidant activities of extracts from olive leaves (EOL). The main contents of EOL were determined by colorimetric methods. The antioxidant activities were assessed by measuring the scavenging free radicals in vitro. To investigate the antioxidant activity in vivo, we detected the survival of Caenorhabditis elegans, under thermal stress. Subsequently the reactive oxygen species (ROS) level, activities of antioxidant enzymes, the expression of HSP-16.2 and the translocation of daf-16 were measured. The results showed that, polyphenols was the main component. EOL could well scavenge DPPH and superoxide anion radicals in vitro. Compared to the control group, the survival rate of C. elegans treated with EOL was extended by 10.43%, under heat stress. The ROS level was reduced, while the expression of hsp-16.2 was increased to protect the organism against the increasing ROS. The level of malondialdehyde (MDA) also decreased sharply. The activities of inner antioxidant enzymes, such as catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GSH-PX) were potentiated, which might have had a correlation with the DAF-16 transcription factor that was induced-turned into the nuclear. Therefore, EOL showed a strong antioxidant ability in vitro and in vivo. Hence, it could be a potential candidate when it came to medicinal and edible plants.

Arbuscular mycorrhizal fungi alleviate arsenic toxicity to Medicago sativa by influencing arsenic speciation and partitioning.

In a pot experiment, Medicago sativa inoculated with/without arbuscular mycorrhizal (AM) fungus Rhizophagus irregularis were grown in four levels (0, 10, 25, and 75 mg/kg) of arsenic (As)-polluted soil to investigate the influences of AM symbiosis on plant As tolerance. The results showed that mycorrhizal inoculation significantly increased plant biomass, while As addition decreased mycorrhizal colonization and hyphal length density. Mycorrhizal inoculation dramatically improved plant phosphorus (P) nutrition, restricted As uptake and retained more As in roots by upregulating the expression of the AM-induced P transporter gene MsPT4 and the metallothionein gene MsMT2. High soil As content downregulated MsPT4 expression. Dimethylarsenic acid (DMA) was detected only in the shoots of mycorrhizal plants, indicating that AM fungi likely play an essential role in As detoxification by biological methylation. The present investigation allowed deeper insights into the As detoxification mechanisms of AM associations and demonstrated the important role of AM fungi in plant resistance under As-contaminated conditions.

Arbuscular mycorrhiza facilitates the accumulation of glycyrrhizin and liquiritin in Glycyrrhiza uralensis under drought stress.

Liquorice (Glycyrrhiza uralensis) is an important medicinal plant for which there is a huge market demand. It has been reported that arbuscular mycorrhizal (AM) symbiosis and drought stress can stimulate the accumulation of the active ingredients, glycyrrhizin and liquiritin, in liquorice plants, but the potential interactions of AM symbiosis and drought stress remain largely unknown. In the present work, we investigated mycorrhizal effects on plant growth and accumulation of glycyrrhizin and liquiritin in liquorice plants under different water regimes. The results indicated that AM plants generally exhibited better growth and physiological status including stomatal conductance, photosynthesis rate, and water use efficiency compared with non-AM plants. AM inoculation up-regulated the expression of an aquaporin gene PIP and decreased root abscisic acid (ABA) concentrations under drought stress. In general, AM plants displayed lower root carbon (C) and nitrogen (N) concentrations, higher phosphorus (P) concentrations, and therefore, lower C:P and N:P ratios but higher C:N ratio than non-AM plants. On the other hand, AM inoculation increased root glycyrrhizin and liquiritin concentrations, and the mycorrhizal effects were more pronounced under moderate drought stress than under well-watered condition or severe drought stress for glycyrrhizin accumulation. The accumulation of glycyrrhizin and liquiritin in AM plants was consistent with the C:N ratio changes in support of the carbon-nutrient balance hypothesis. Moreover, the glycyrrhizin accumulation was positively correlated with the expression of glycyrrhizin biosynthesis genes SQS1, ß-AS, CYP88D6, and CYP72A154. By contrast, no significant interaction of AM inoculation with water treatment was observed for liquiritin accumulation, while we similarly observed a positive correlation between liquiritin accumulation and the expression of a liquiritin biosynthesis gene CHS. These results suggested that AM inoculation in combination with proper water management potentially could improve glycyrrhizin and liquiritin accumulation in liquorice roots and may be practiced to promote liquorice cultivation.

[Experimental study on Rhizoma sparganii and Radices zedoariae treating hepatic fibrosis].

OBJECTIVE: To study the effects of rhizoma sparganii and radices zedoariae on hepatic fibrosis. METHOD: The rat immunohepatic fibrosis model was made by intraperitoneal injection of porcine serum and treated with rhizoma sparganii and radices zedoariae. The ALT, GGT, TP, ALb, A/G, IVC, LN, HA and the pathological change of the liver were observed. RESULT: Rhizoma sparganii and radices zedoariae could increase TP, ALb, A/G, decrease ALT, GGT, IVC, LN, HA and improve the pathological change. CONCLUSION: Rhizoma sparganii and radices zedoariae can protect hepatic cells, alleviate degeneration and necrosis, recover structure and function, and reduce the proliferation of fibrous tissue.