Network pharmacology combined with experimental validation to investigate the effect of Rongjin Niantong Fang on chondrocyte apoptosis in knee osteoarthritis.

Knee osteoarthritis (KOA) is a chronic degenerative disease that affects the quality of life of middle­aged and elderly individuals, and is one of the major factors leading to disability. Rongjin Niantong Fang (RJNTF) can alleviate the clinical symptoms of patients with KOA, but the molecular mechanism underlying its beneficial effects on KOA remains unknown. Using pharmacological analysis and in vitro experiments, the active components of RJNTF were analyzed to explore their potential therapeutic targets and mechanisms in KOA. The potential targets and core signaling pathways by which RJNTF exerts its effects on KOA were obtained from databases such as Gene Expression Omnibus, Traditional Chinese Medicine Systems Pharmacology and Analysis Platform. Subsequently, chondrocyte apoptosis was modeled using hydrogen peroxide (H2O2). Cell Counting Kit­8 assay involving a poly [ADP­ribose] polymerase­1 (PARP1) inhibitor, DAPI staining, reverse transcription­quantitative PCR, Annexin V­FITC/PI staining and flow cytometry, western blotting and co­immunoprecipitation analysis were used to determine the therapeutic efficacy of RJNTF on KOA and to uncover the molecular mechanism. It was found that PARP1­knockdown lentivirus, incubation with PARP1 inhibitor PJ34, medium and high doses of RJNTF significantly reduced H2O2­induced chondrocyte apoptosis. Medium and high doses of RJNTF downregulated the expression of cleaved caspase­3, cleaved PARP1 and PAR total proteins, as well as nucleus proteins of apoptosis­inducing factor (AIF) and migration inhibitory factor (MIF), and upregulated the expression of caspase­3, PARP1 total protein, as well as the cytoplasmic expression of AIF and MIF, suggesting that RJNTF may inhibit chondrocyte apoptosis through the PARP1/AIF signaling pathway.

Strategies to attenuate ciprofloxacin inhibition on enhanced biological phosphorus removal from wastewater and its recoverability.

The inhibiting effects of ciprofloxacin (CIP) on enhanced biological phosphorus removal (EBPR) were investigated with no change in reactor operation and with increased aeration rate and sludge retention time (SRT) to explore inhibition-alleviating solutions. Additionally, performance recoverability was evaluated. The results showed that the phosphorus removal efficiency in the presence of 0.002-0.092 mg/L CIP for 7 days was only 12.5%. Increasing the aeration rate relieved inhibition (33.5% phosphorus removal efficiency on Day 7), and increasing SRT slowed EBPR performance deterioration. The EBPR performance recovered from CIP inhibition and increases in the aeration rate and SRT resulted in different recovery phenomena. The maximum PO43--P release rate continued to decrease in the first 2 days of the recovery stage and then gradually increased. However, the maximum PO43--P uptake rate immediately increased at different rates among reactors, which might be attributed to variations in the microbial community structure, decreased poly-P content, and enhanced abundances of ABC transporters and quorum sensing. It was found that some microorganisms associated with phosphorus removal were more tolerant to CIP than glycogen accumulating organisms. Moreover, the increased relative abundance of the qepA gene indicated that the microorganisms in the EBPR system had strong antibiotic resistance capacity. The bacterial community structure was significantly affected by CIP and could not recover to the initial structure. The results help to provide technical support for the operation of the EBPR process in the presence of CIP and to increase the understanding of system recoverability.

Selenium intake and multiple health-related outcomes: an umbrella review of meta-analyses.

Selenium is an essential trace metalloid element that is associated with fundamental importance to human health. Our umbrella review aimed to evaluate the quality of evidence, validity, and biases in the relationship between selenium intake and health-related outcomes according to published systematic reviews with pooled data and meta-analyses. Selenium intake is associated with a decreased risk of digestive system cancers, all-cause mortality, depression, and Keshan disease, when in children reduce the risk of Kashin-Beck disease. Additionally, selenium supplementation can improve sperm quality, polycystic ovary syndrome, autoimmune thyroid disease, cardiovascular disease, and infective outcomes. Selenium supplementation also has relationship with a decreased concentration of serum lipids including total cholesterol and very low-density lipoprotein cholesterol. However, no evidence has shown that selenium is associated with better outcomes among patients in intensive care units. Furthermore, selenium intake may be related with a higher risk of type 2 diabetes and non-melanoma skin cancers. Moreover, most of included studies are evaluated as low quality according to our evidence assessment. Based on our study findings and the limited advantages of selenium intake, it is not recommended to receive extra supplementary selenium for general populations, and selenium supplementation should not be continued in patients whose selenium-deficient status has been corrected.

Mitochondria-anchoring self-assembled nanoparticles for multi-path energy depletion: A "nano bomb" in chemo-co-starvation therapy.

As the main systemic treatment for triple-negative breast cancer (TNBC), the bleak medical prognosis of chemotherapy resulted in impaired life quality by tumor recurrence and metastasis. The feasible cancer starvation therapy could inhibit tumor progression by blocking energy supplements, however, the mono-therapeutic modality showed limited curing efficacy due to heterogeneity and abnormal energy metabolism of TNBC. Thus, the development of a synergistic nano-therapeutic modality involving different anti-tumor mechanisms to simultaneously transport medicines to the organelle where metabolism took place, might remarkably improve curing efficacy, targeting ability, and bio-safety. Herein, the hybrid BLG@TPGS NPs were prepared by doping multi-path energy inhibitors Berberine (BBR) and Lonidamine (LND) as well as the chemotherapeutic agent Gambogic acid (GA). Our research indicated that Nanobomb-BLG@TPGS NPs inherited the mitochondria targeting ability from BBR to accumulate precisely at the "energy factory" mitochondria, and then induce starvation therapy to efficiently eradicated cancer cells by coordinately powered off tumor cells via a "three-prone strategy" to cut off mitochondrial respiration, glycolysis, and glutamine metabolism. The inhibition of tumor proliferation and migration was enlarged by the synergistic combination with chemotherapy. Besides, apoptosis via mitochondria pathway and mitochondria fragmentation supported the hypothesis that NPs eliminated MDA-MB-231 cells by violently attacking MDA-MB-231 cells and especially the mitochondria. In summary, this synergistic chemo-co-starvation nanomedicine proposed an innovative site-specific targeting strategy for improved tumor treatment and decreased toxicity to normal tissues, which provided an option for clinical TNBC-sensitive treatment.

Iron intake and multiple health outcomes: Umbrella review.

Iron is an essential trace element, while excess iron can lead to different levels of physical abnormalities or diseases. This umbrella review aimed to conduct a systematic evaluation of the possible relationships between iron intake and various health outcomes. We retrieved PubMed, Embase, Web of Science, Scopus, and the Cochrane Database of Systematic Reviews from inception through May 2021. A total of 34 meta-analyses with 46 unique health outcomes were identified. Heme iron intake was positively associated with nine outcomes, including colorectal cancer, type 2 diabetes mellitus, and cardiovascular disease mortality, while dietary total iron intake could decrease the risk of colorectal adenoma, esophageal cancer, coronary heart disease, and depression. Iron supplementation was a protective factor against eight outcomes. However, it was associated with decreased length and weight gain. The quality of evidence for most outcomes was "low" or "very low" with the remaining eleven as "high" or "moderate". All outcomes were categorized as class III, IV, or NS based on evidence classification. Although high iron intake has been identified to be significantly associated with a range of outcomes, firm universal conclusions about its beneficial or negative effects cannot be drawn given the low quality of evidence for most outcomes.

Zinc Intakes and Health Outcomes: An Umbrella Review.

It is widely accepted that the zinc element is crucial in human beings. Zinc has gained more attention during the COVID-19 pandemic due to its utilization for the treatment and prevention of respiratory tract infections. However, some studies also pointed out that zinc intake might cause unwanted side effects and even be dangerous when overdosed. To reveal the relationship between zinc intake and health outcomes, we performed an umbrella review from human studies. In total, the umbrella review included 43 articles and identified 11 outcomes for dietary zinc intake and 86 outcomes for supplementary zinc intake. Dietary zinc intake in the highest dose would decrease the risk of overall and specific digestive tract cancers, depression, and type 2 diabetes mellitus (T2DM) in adults. Supplementary zinc consumption in adults was linked to an improvement of depression, antioxidant capacity and sperm quality, higher serum zinc concentration, and lower concentration of inflammatory markers. Zinc supplementation in children would reduce the incidence of diarrhea and pneumonia, improve zinc deficiency and boost growth. However, zinc might not decrease all-cause mortality in adults or the in-hospital mortality of COVID-19. And better maternal and neonatal outcomes may not derive from pregnant women who consumed higher or lower doses of zinc supplementation (>20 mg/day and <20 mg/day, respectively). Dose-response analyses revealed that a daily 5 mg increment of zinc would lower the risk of colorectal and esophageal cancer, whereas a large dose of zinc supplementation (daily 100 mg) showed no benefit in reducing prostate cancer risk.

Serum Metabolic Profiling Analysis of Gout Patients Treated with Traditional Chinese Medicine Tongfengtai Granules Based on Gas Chromatography-Mass Spectrometry.

Gout has become a public health problem that seriously threatens human health. Traditional Chinese medicines (TCMs) have a long history of treating gout and have some advantages compared with the conventional medicines. Compound TCM Tongfengtai granules are gradually being used for clinical treatment of gout, but its mechanism is still unclear. The purpose of this study was to explore the metabolic profiling of serum from gout patients before and after treatment with Tongfengtai granules and identify the differential metabolites and related metabolic pathways. A total of 40 gout patients hospitalized in Shenzhen Traditional Chinese Medicine Hospital from 2018 to March 2019 were recruited in the current study, and serum samples from these patients before and after treatment with Tongfengtai granules were collected. Gas chromatography-mass spectrometry (GC-MS) assay was used to identify serum metabolites. The OPLS-DA VIP method was used to screen for potential metabolic biomarkers, and MetaboAnalyst 4.0 was used to identify related metabolic pathways. The result showed that there was a significant difference in the concentrations of six metabolites in the serum after treatment: D-galactose, lactic acid, 3-hydroxybutyric acid, D-pyran (type) glucose, alanine, and L-isoleucine. Except D-pyran (type) glucose, the serum concentrations of the other five metabolites were all significantly reduced. Besides, pathway enrichment analysis found that these potential metabolic biomarkers were mainly involved in lactose degradation and the glucose-alanine cycle. Thus, the serum metabolic profiling of gout patients treated with Tongfengtai granules changed, and the differential metabolites and related metabolic pathways might provide clues for understanding the mechanism of Tongfengtai granules.

Phenylethanoid glycosides of Callicarpa kwangtungensis Chun exert cardioprotective effect by weakening Na+-K+-ATPase/Src/ERK1/2 pathway and inhibiting apoptosis mediated by oxidative stress and inflammation.

ETHNOPHARMACOLOGICAL RELEVANCE: Callicarpa kwangtungensis Chun (C. kwangtungensis) is a very famous herbal medicine with the function of promoting blood circulation and removing blood stasis which is beneficial for cardiovascular disease (CVD). Phenylethanoid glycosides (PGs) are the major class of active ingredients in C. kwangtungensis and present significant anti-oxidative and anti-inflammatory property related to apoptosis. Therefore, this study aimed to investigate the effects of total phenylethanoid glycosides of C. kwangtungensis (CK-PGs) on isoproterenol (ISO) induced myocardial ischemic injury (MI) and the mechanisms related to the apoptosis mediated by oxidative damage and inflammation. METHODS: The myocardial ischemia animal model was established as subcutaneous injecting ISO. Echocardiography and biomarkers were employed to determine the degree of myocardial damage. Histopathological changes were observed by hematoxylin and eosin test. The TUNEL staining and activity of caspase-3 were measured to detect the level of apoptosis which is medicated by the oxidative damage detected by the level of MDA, GSH and ROS tested with the kit and the inflammation reflected by TNF-α. The activity of Na+-K+-ATPase (NKA) was detected by the commercial kits, whose expression was measured by immunohistochemistry analysis. At last, Western blot analysis was used to measure Na+-K+-ATPase/Src/ERK1/2 and Bax/Bcl-2 pathway. RESULTS: CK-PGs showed cardioprotective effect against ISO-induced myocardial ischemic injury evidenced by improving heart function and lowering myocardial injury markers. CK-PGs could inhibit the level of apoptosis as shown by the decrease of the TUNEL-positive cells, the activity of caspase-3 and increase of the expression of Bax. CK-PGs also reduced oxidative stress and inflammation to suppress apoptosis by decreasing the level of ROS, MDA, and increasing GSH activity and lowering the level of TNF-α. In addition, CK-PGs exerted the protection by increasing the activity and the expression of NKA. Meanwhile, Na+-K+-ATPase/Src/ERK1/2pathway was weakened for the inhibition of apoptosis. CONCLUSIONS: CK-PGs could protect cardiomyocytes from myocardial injury through suppressing Na+-K+-ATPase/Src/ERK1/2 pathway and inhibiting apoptosis mediated by oxidative stress and inflammation.

Self-Delivered and Self-Monitored Chemo-Photodynamic Nanoparticles with Light-Triggered Synergistic Antitumor Therapies by Downregulation of HIF-1α and Depletion of GSH.

Photodynamic therapy (PDT), a clinically approved cancer treatment, has faced many drawbacks that restricted its applications. For example, the hypoxia-induced elevated hypoxia-inducible factor-1α (HIF-1α) may desensitize tumors to PDT, and the high concentration of glutathione (GSH) in cancer cells can also neutralize the generated reactive oxygen species (ROS) during PDT, resulting in insufficient therapy. Moreover, extra probes for imaging-guided visualization therapy are always needed to track drug release or distribution, while it may decrease the drug loading of the drug delivery system (DDS). In the present study, we have designed and prepared a novel multifunctional combined therapy nanoparticle (ZnPc@Cur-S-OA NPs), in which curcumin (Cur) was not only used as a chemotherapy drug to achieve a combination therapy with PDT via downregulating HIF-1α and depleting GSH in B16F10 cells but also designed as a small-molecule ROS-triggered release prodrug to deliver the photosensitizer (PS). The red fluorescence of PS in the nanoparticles (NPs) can be used to track the NPs distribution, while the green fluorescence of Cur showed an "OFF-ON" activation, which enables additional imaging and real-time self-monitoring capabilities. These results proved that the prepared combined therapy NPs were more effective to inhibit the growth of B16F10 mouse melanoma tumor than was monotherapy without eliciting systemic toxicity either in vitro or in vivo, which indicated the combined therapy NPs as an effective way to improve the PDT efficacy via downregulation of HIF-1α and depletion of GSH. Thus, the strategy of using a multifunctional natural product as the stimuli-responsive carrier as well as the synergist with PDT for enhancing antitumor efficacy via multiple pathways may open an alternative avenue to fabricate new self-delivery combination therapy nanodrugs. Besides, the fluorescence emitted from the drug can be used for real-time self-monitoring of drug release and distribution, which has great potential in clinic to adjust the administration dose and irradiation time for different tumor types and stages for individual therapy.

The acute effects of erythromycin and oxytetracycline on enhanced biological phosphorus removal system: shift in bacterial community structure.

Since extensive application, an increasing amount of antibiotics has been released into wastewater treatment plants. In this study, the enhanced biological phosphorus removal (EBPR) system was fed with synthetic wastewater containing erythromycin (ERY) and oxytetracycline (OTC) for 7 days to evaluate the variations of solution ortho-P (SOP), volatile fatty acid (VFA), poly-bhydroxyalkanoates (PHAs), specific oxygen uptake rater (SOUR), and microbial community in the EBPR system. The obtained results showed that the P-removal efficiency decreased to 0.0%, and at the end of the experiment, only less than 20% of the VFA could be consumed. Besides, the variable processes of P and PHAs were destroyed. Moreover, to better grasp the inhibitory mechanism of antibiotics, microbial community compositions of activated sludge sampled in all reactors were investigated by high-throughput sequencing techniques. Results of comparative and evolutionary analysis revealed that high concentrations (5 and 10 mg/L) of ERY and OTC could seriously shift microbial communities, while combined antibiotics could induce more. Additionally, Accumulibacter and Competibacter were two primary microorganisms at the genus level in the EBPR system. Accumulibacter decreased seriously for exposure to antibiotics, while Competibacter increased in all experimental reactors especially in combined antibiotics reactor.

Understanding the performance of microbial community induced by ZnO nanoparticles in enhanced biological phosphorus removal system and its recoverability.

In this study, the impacts of ZnO Nanoparticles (NPs) on the microbial community in enhanced biological phosphorus removal (EBPR) system and its recoverability were investigated. High-throughput sequencing was applied to study the microbial community shift. Results show that the species richness in the EBPR system was reduced under the condition of ZnO NPs with high concentration (above 6mg/L). Evolution analysis suggests that higher concentration ZnO NPs induced more microbial community shift. According to the analysis on genus level, Competibacter was more impressionable than Accumulibacter after exposure to 2mg/L ZnO NPs. Nonetheless, this phenomenon could not be found as the concentration of ZnO NPs got higher (above 6mg/L). Accumulibacter could reach to the initial level after recover for 20days, whereas Competibacter could not recover even when the concentration of ZnO NPs was only 2mg/L. Interestingly, although the phosphorus removal (P-removal) process was re-achieved, the microbial community in reactors was irreversible.

Short-term performance of enhanced biological phosphorus removal (EBPR) system exposed to erythromycin (ERY) and oxytetracycline (OTC).

The effects of Erythromycin (ERY) and oxytetracycline (OTC), including individual and combinative effect, on enhanced biological phosphorus removal (EBPR) system within a short-term (24h) were evaluated in this study. Results showed that the P-removal efficiency decreased to 34.6% and 0.0% under the effect of ERY (10mg/L) and OTC (10mg/L) for 24h. OTC concentration higher than 5mg/L was sufficient to cause serious adverse impact on the EBPR performance. While the performance of EBPR system will be impacted by ERY above 10mg/L. OTC, due to its special antibacterial action to the gram-negative bacteria which most PAOs belong to, has more serious negative effect on the EBPR performance than ERY does. Moreover, in the combined antibiotics test, neither synergistic nor antagonistic effect was detected between ERY and OTC. Finally, ERY (10mg/L) and OTC (10mg/L) could inhibit the microorganisms' activity, while couldn't induce serious microorganisms death within 24h.

Understanding the impact of influent nitrogen concentration on granule size and microbial community in a granule-based enhanced biological phosphorus removal system.

To better understand the effect of influent nitrogen concentration on granule size and microbial community in a granule-based enhanced biological phosphorus removal system, three influent nitrogen concentrations were tested while carbon concentration was an unlimited factor. The results show that although ammonium and phosphate were well removed in the tested nitrogen concentration range (20-50 mg L(-1)), granule size, the amount of phosphate accumulating organisms (PAOs) and microbial activity were affected significantly. A possible mechanism for the effect of influent nitrogen concentration on granule size is proposed based on the experimental results. The increase in proteins/polysaccharides ratio caused by high influent nitrogen concentration plays a crucial role in granule breakage. The small granule size then weakens simultaneous nitrification-denitrification, which further causes higher nitrate concentration in the effluent and lower amount of PAOs in sludge. Consequently, phosphate concentration in the anaerobic phase decreases, which plays the secondary role in granule breakage.

[Ecological effect of recirculated landfill leachate with different hydraulic loading on plant-soil system].

The ecological effect of irrigating recirculated leachate with different hydraulic loading on bermudagrass (Cynodon dactylon) and soil system was evaluated. The results show that leachate irrigation with hydraulic loading of 2.77-12.00 mm d(-1) leads to high chlorophyll content, low proline (Pro) content, as well as low malondialdehyde (MDA) and H2O2 content. Soil enzyme activities, respirations, microbial biomass and ratio of biomass carbon to organic carbon (Cmic/Corg) are rather higher. Among leachte irrigation group, leachate irrigation with hydraulic loading of 6.46-10.15 mm x d(-1) leads to declined proline, increased chlorophyll content, low POD activity and low content of MDA and H2O2. Soil bio-activity related parameters, such as soil enzyme activities, respirations, microbial biomass and Cmic/Corg are all enhanced at 6.46-10.15 mm x d(-1) of hydraulic loading. However, when hydraulic loading increases to 12.00 mm x d(-1) or declines to 2.77-4.16 mm x d(-1), the stress of irrigation on bermudagrass is aggravated, and soil bioactivity declines. The results reveal that leachate irrigation could alleviate the stress of environment on bermudagrass and improve the bio-activity of soil. The positive effect of leachate irrigation on the plant-soil system might be contributed to changes of soil water and physico-chemical property after leachate irrigated to the soil. The experimental results suggest that leachate irrigation could benefit plant-soil system, especially when controlled at suitable hydraulic loading.

[Responses of antioxidation system of Cynodon dactylon to recirculated landfill leachate irrigation].

With pot experiment, this paper studied the membrane lipid peroxidation and the variations of antioxidation system in Cynodon dactylon under recirculated landfill leachate irrigation. The results showed that when irrigated with low dilution ratio (< 25%) leachate, the chlorophyll a/b ratio increased with increasing dilution ratio, membrane permeability and MDA and H2O2 contents were in adverse, and membrane lipid peroxidation was relatively weak. However, with the increasing leachate dilution ratio (> 25%), there existed an obvious negative fect on Cynodon dactylon, i.e., the chlorophyll a/b ratio decreased, while cell membrane permeability and MDA and H2O2 contents increased, which meant that the membrane lipid peroxidation was accelerated. The contents antioxidants AsA, GSH and Car also showed the similar trend, i.e., they increased with increasing leachate dilution ratio when irrigated with low dilution ratio leachate, but decreased under medium or high dilution ratio leachate irrigation. Among three test anti-oxidative enzymes, SOD and POD activities showed a similar change test antioxidants, and POD activity was more sensitive, while CAT activity was on the contrary. The contents test antioxidants and the activities of SOD and POD were negatively and significantly correlated to MDA content, indicating that they might play an important role in preventing Cynodon dactylon from cell membrane lipid peroxdation.

[Studies on organ phytomorphology of leaves from Rehmannia glutinosa].

OBJECTIVE: To set up the fittest medium and optimum condition for leaves from Rehmannia glutinosa to form regenerated plantlets directly and to induce microtubers. METHOD: Leaves of different phases obtained from the virus-free plantlets were incubated in some kinds of media on the back side. RESULT AND CONCLUSION: The fittest medium for the formation of regenerated plantlet was MS + 6-BA 2.0 mg.L-1 and the fittest medium for the induction of microtuber was MS + 6-BA 1.0 mg.L-1 + NAA 0.5 mg.L-1. The optimum incubation temperaue was 25 degrees C and the light length was (2,000-3,000 lx) 12 h.d-1.