Qualitative and quantitative analysis of chemical components in Qianggan capsule by UHPLC-Q-TOF-MS/MS and LC-sMRM.

Qianggan capsule (QGC) is a complex preparation composed of 16 traditional Chinese medicines (TCM) that can clear heat and dampness, fortify the spleen and blood, typify qi and relieve depression. However, the chemical composition of QGC remains incompletely understood, despite its clinical use in treating chronic hepatitis and liver injury. The objective of this study was to explore the quality markers of QGC through qualitative and quantitative analysis of its chemical components. First, the chemical composition of QGC was qualitatively analyzed using UHPLC-Q-TOF-MS/MS. Subsequently, the LC-sMRM method was developed and optimized to accurately quantify various chemical components of 10 batches of QGC. Finally, the variations in chemical components between batches were analyzed via multivariate statistical analysis. UHPLC-Q-TOF-MS/MS analysis revealed 167 chemical constituents in QGC, comprised of 48 flavonoids, 32 terpenoids, 18 phenolic acids, 9 coumarins, 9 phenylpropanoids, and 51 nucleosides, sugars, amino acids, anthraquinones, and other compounds. The LC-sMRM method was established for the quantitative analysis of 42 chemical components in 10 batches of QGC. The ultrasonic-assisted extraction parameters were optimized using RSM. Compared with conventional MRM, sMRM demonstrated superior sensitivity and precision. PCA and OPLS-DA identified eight chemical components with content differences among batches. This study established the chemical composition of QGC, offering useful guidance for assessing its quality.

Metabolic activation and cytochrome P450 inhibition of piperlonguminine mediated by CYP3A4.

Piperlonguminine (PLG) is a major alkaloid found in Piper longum fruits. It has been shown to possess a variety of biological activities, including anti-tumor, anti-hyperlipidemic, anti-renal fibrosis and anti-inflammatory properties. Previous studies have reported that PLG inhibits various CYP450 enzymes. The main objective of this study was to identify reactive metabolites of PLG in vitro and assess its ability to inhibit CYP450. In rat and human liver microsomal incubation systems exposed to PLG, two oxidized metabolites (M1 and M2) were detected. Additionally, in microsomes where N-acetylcysteine was used as a trapping agent, N-acetylcysteine conjugates (M3, M4, M5 and M6) of four isomeric O-quinone-derived reactive metabolites were found. The formation of metabolites was dependent on NADPH. Inhibition and recombinant CYP450 enzyme incubation experiments showed that CYP3A4 was the primary enzyme responsible for the metabolic activation of PLG. This study characterized the O-dealkylated metabolite (M1) through chemical synthesis. The IC50 shift assay showed time-dependent inhibition of CYP3A4, 2C9, 2E1, 2C8 and 2D6 by PLG. This research contributes to the understanding of PLG-induced enzyme inhibition and bioactivation.

Rapid visual detection of sulfur dioxide residues in food using acid-sensitive CdTe quantum dots-loaded alginate hydrogel beads.

Acid-sensitive CdTe quantum dots-loaded alginate hydrogel (CdTe QDs-AH) beads were designed for the visual detection of SO2 residues. As proof of concept, two types of CdTe QDs were selected as model probes and embedded in AH beads. The entire test was performed within 25 min in a modified double-layer test tube with one bead fixed above the sample solution. Adding citric acid and heating at 70 ℃ for 20 min transformed the sulfites in the solution into SO2 gas, which then quenched the fluorescence of the CdTe QDs-AH beads. Using this assay, qualitative, naked-eye detection of SO2 residues was achieved in the concentration range of 25-300 ppm, as well as precise quantification was possible based on the difference in the average fluorescence brightness of the beads before and after the reaction. Five food types were successfully analysed using this method, which is simpler and more economical than existing methods, and does not require complex pretreatment.

Metabolic engineering of glycolysis in Escherichia coli for efficient production of patchoulol and τ-cadinol.

Glucose metabolism suppresses the microbial synthesis of sesquiterpenes with a syndrome of "too much of a good thing can be bad". Here, patchoulol production in Escherichia coli was increased 2.02 times by engineering patchoulol synthase to obtain an initial strain. Knocking out the synthetic pathway for cyclic adenosine monophosphate relieved glucose repression and improved patchoulol titer and yield by 27.7 % and 43.1 %, respectively. A glycolysis regulation device mediated by pyruvate sensing was constructed which effectively alleviated overflow metabolism in a high-glucose environment with 10.2 % greater patchoulol titer in strain 070QA. Without fine-tuning the glucose-feeding rate, patchoulol titer further increased to 1675.1 mg/L in a 5-L bioreactor experiment, which was the highest level reported in E. coli. Using strain 070QA as a chassis, the τ-cadinol titer reached 15.2 g/L, representing the first report for microbial production of τ-cadinol. These findings will aid in the industrial production of sesquiterpene.

A virtual reality platform to evaluate the effects of supernumerary limbs' appearance.

Supernumerary robot limbs (SL) can expand the ability of users by increasing the number of degrees of freedom that they control. While several SLs have been designed and tested on human participants, the effect of the limb's appearance on the user's acceptance, embodiment and device usage is not yet understood. We developed a virtual reality platform with a three-arm avatar that enabled us to systematically investigate the effect of the supernumerary limb's appearance on their perception and motion control performance. A pilot study with 14 participants exhibited similar performance, workload and preference in human-like or robot-like appearance with a trend of preference for the robotic appearance.

Using Inducible Osteoblastic Lineage-Specific Stat3 Knockout Mice to Study Alveolar Bone Remodeling During Orthodontic Tooth Movement.

The alveolar bone, with a high turnover rate, is the most actively-remodeling bone in the body. Orthodontic tooth movement (OTM) is a common artificial process of alveolar bone remodeling in response to mechanical force, but the underlying mechanism remains elusive. Previous studies have been unable to reveal the precise mechanism of bone remodeling in any time and space due to animal model-related restrictions. The signal transducer and activator of transcription 3 (STAT3) is important in bone metabolism, but its role in osteoblasts during OTM is unclear. To provide in vivo evidence that STAT3 participates in OTM at specific time points and in particular cells during OTM, we generated a tamoxifen-inducible osteoblast lineage-specific Stat3 knockout mouse model, applied orthodontic force, and analyzed the alveolar bone phenotype. Micro-computed tomography (Micro-CT) and stereo microscopy were used to access OTM distance. Histological analysis selected the area located within three roots of the first molar (M1) in the cross-section of the maxillary bone as the region of interest (ROI) to evaluate the metabolic activity of osteoblasts and osteoclasts, indicating the effect of orthodontic force on alveolar bone. In short, we provide a protocol for using inducible osteoblast lineage-specific Stat3 knockout mice to study bone remodeling under orthodontic force and describe methods for analyzing alveolar bone remodeling during OTM, thus shedding new light on skeletal mechanical biology.

Insights into rapidly recovering the autotrophic nitrogen removal performance of single-stage partial nitritation-anammox systems: Reconstructing granular sludge and its functional microbes synergy.

Partial nitritation-anammox (PNA) deteriorates easily and is difficult to recover. After an airlift inner-circulation partition bioreactor was impacted by low NH4+-N wastewater containing organic matter, Nitrospira and Denitratisoma propagated rapidly, granular sludge disintegrated, and the total nitrogen removal efficiency (TNRE) decreased from 68.27 % to 5.97 %. This study used a unique strategy to recover deteriorated single-stage PNA systems and explored the mechanism of rapid performance recovery. The TNRE of the system recovered up to 61.77 % in 43 days. The high nitrogen loading rate and hydraulic shear force from the airlift caused the sludge in the reactor to granulate again. The microbial community structure recovered, with a decrease in the abundance of Nitrospira (0.05 %) and enrichment of Candidatus Brocadia (8.82 %). A favorable synergy among functional microbes in the reactor was thus re-established, promoting the rapid recovery of the nitrogen removal performance. This study provides a feasible recovery strategy for PNA processes.

BmCDK5 Affects Cell Proliferation and Cytoskeleton Morphology by Interacting with BmCNN in Bombyx mori.

The ordered cell cycle is important to the proliferation and differentiation of living organisms. Cyclin-dependent kinases (CDKs) perform regulatory functions in different phases of the cell cycle process to ensure order. We identified a homologous gene of the Cyclin-dependent kinase family, BmCDK5, in Bombyx mori. BmCDK5 contains the STKc_CDK5 domain. The BmCDK5 gene was highly expressed in S phase. Overexpression of the BmCDK5 gene accelerates the process of the cell cycle's mitotic period (M) and promotes cell proliferation; knocking out the BmCDK5 gene inhibited cell proliferation. Furthermore, we identified a protein, BmCNN, which can interact with BmCDK5 and represents the same express patterns as the BmCDK5 gene in the cell cycle phase and the spatial-temporal expression of B. mori. This study revealed that BmCDK5 and BmCNN play roles in promoting cell proliferation and regulating cytoskeleton morphology, but do not induce expression changes in microtubule protein. Therefore, our findings provide a new insight; the BmCDK5 gene has a regulatory effect on the cell cycle and proliferation of B. mori, which is presumably due to the interaction between BmCDK5 and BmCNN regulating changes in the cytoskeleton.

Response surface model comparison and combinations for remifentanil and propofol in describing response to esophageal instrumentation and adverse respiratory events.

BACKGROUND: The primary aim of this essay was to explore the best fitting model in remifentanil-propofol combined administrations during esophageal instrumentation (EI) from five distinct response surface models. The secondary aim was to combine the models to give appropriate effect-site drug concentrations (Ces) range with maximal comfort and safety. METHODS: The Greco, reduced Greco, Minto, Scaled C50 Hierarchy and Fixed C50 Hierarchy models were constructed to fit four drug effects: loss of response to esophageal instrumentation (LREI), loss of response to esophageal instrumentation revised (LREIR), intolerable ventilatory depression (IVD) and respiratory compromise (RC). Models were tested by chi-square statistical test and evaluated with Akaike Information Criterion (AIC). Model prediction performance were measured by successful prediction rate (SPR) and three prediction errors. RESULTS: The reduced Greco model was the best fitting model for LREI and RC, and the Minto model was the best fitting model for LREIR and IVD. The SPRs of reduced Greco model for LREI and RC were 81.76% and 79.81%. The SPRs of Minto model for LREIR and IVD were 80.32% and 80.12%. Overlay of the reduced Greco model for LREI and Minto model for IVD offered visual aid for guidance in drug administration. CONCLUSION: Using proper response surface model to fit different drug effects will describe the interactions between anesthetic drugs better. Combining response surface models to select the more reliable effect-site drug concentrations range can be used to guide clinical drug administration with greater safety and provide an improvement of anesthesia precision.

Current understandings and clinical translation of nanomedicines for breast cancer therapy.

Breast cancer is one of the most frequently diagnosed cancers that is threatening women's life. Current clinical treatment regimens for breast cancer often involve neoadjuvant and adjuvant systemic therapies, which somewhat are associated with unfavorable features. Also, the heterogeneous nature of breast cancers requires precision medicine that cannot be fulfilled by a single type of systemically administered drug. Taking advantage of the nanocarriers, nanomedicines emerge as promising therapeutic agents for breast cancer that could resolve the defects of drugs and achieve precise drug delivery to almost all sites of primary and metastatic breast tumors (e.g. tumor vasculature, tumor stroma components, breast cancer cells, and some immune cells). Seven nanomedicines as represented by Doxil® have been approved for breast cancer clinical treatment so far. More nanomedicines including both non-targeting and active targeting nanomedicines are being evaluated in the clinical trials. However, we have to realize that the translation of nanomedicines, particularly the active targeting nanomedicines is not as successful as people have expected. This review provides a comprehensive landscape of the nanomedicines for breast cancer treatment, from laboratory investigations to clinical applications. We also highlight the key advances in the understanding of the biological fate and the targeting strategies of breast cancer nanomedicine and the implications to clinical translation.

Advances in Nanodelivery of Green Tea Catechins to Enhance the Anticancer Activity.

Cancer is one of the leading causes of death globally. A variety of phenolic compounds display preventative and therapeutic effects against cancers. Green teas are rich in phenolics. Catechins are the most dominant phenolic component in green teas. Studies have shown that catechins have anticancer activity in various cancer models. The anticancer activity of catechins, however, may be compromised due to their low oral bioavailability. Nanodelivery emerges as a promising way to improve the oral bioavailability and anticancer activity of catechins. Research in this area has been actively conducted in recent decades. This review provides the molecular mechanisms of the anticancer effects of catechins, the factors that limit the oral bioavailability of catechins, and the latest advances of delivering catechins using nanodelivery systems through different routes to enhance their anticancer activity.

Predator odor exposure increases social contact in adolescents and parental behavior in adulthood in Brandt's voles.

Research suggests that predation risk during adolescence can program adult stress response and emotional behavior; however, little is known about the short-term and lasting residual effects of this experience on social behavior. We explored this concept in social Brandt's voles (Lasiopodomys brandtii). Adolescent male and female voles were exposed to distilled water, rabbit urine (as a non-predator stimulus), and cat urine for 60 min daily from postnatal day (PND) 28-49. Social play tests were conducted immediately following exposure on PND 28, 35, 42, and 49. In the social play test, repeated cat odor (CO) exposure enhanced the contact behavior of voles with their cagemate. Adolescent exposure to CO did not affect behavioral responses toward unrelated pups in the alloparental behavior test or same-sex individuals in the social interaction test. However, exposure to CO significantly enhanced the licking/grooming behavior of voles towards their own pups in the home cage parental behavior test. Repeated CO exposure significantly inhibited weight gain in male voles during adolescence. This effect was transmitted to the next generation, with lower weight gain in offspring before weaning. Following repeated CO exposure, males tended to have more female offspring whereas females produced more offspring, suggesting an adaptive strategy to increase inclusive fitness under predatory risk. These findings demonstrate that adolescent exposure to predatory risk augments adolescent social contact and adult parental behavior and suggest a role for improved inclusive fitness in mediating long-term outcomes.

Psychological distress and sleep quality of COVID-19 patients in Wuhan, a lockdown city as the epicenter of COVID-19.

The major Corona Virus Disease 2019 (COVID-19) outbreak caused tens of thousands of diagnosed patients quarantined and treated in designated hospitals in Wuhan, the epicenter of the disease in China. Evidence for the psychological problems of COVID-19 patients was limited. Here we report a cross-sectional study of the mental distress and sleep quality of patients in a single center in Wuhan. The study was based on a combined questionnaire of basic questions designed by the study group, Self-Rating Anxiety Scale (SAS), Self-Rating Depression Scale (SDS), and Pittsburgh Sleep Quality Index (PSQI). On Feb 17th and Mar 14th, two groups of patients were recruited respectively in a designated hospital for COVID-19. Univariate analysis and regression models were used to identify predictors for patients' psychological distress and sleep quality. In total, there were 202 participants in our combined sample. The average SAS, SDS, and PSQI score of participants were 44.2, 51.7, and 9.3 respectively. Factors associated with SAS score include gender, subjective evaluation of disease symptoms, and evaluation of medical staffs' attitude. Gender, age, education level, frequency of contacting with family, subjective knowledge level of COVID 19, and evaluation of medical staffs' attitude are associated with participants SDS score. Factors associated with PSQI score are age and subjective evaluation of disease symptoms.

Immunogenicity and protective efficacy induced by an mRNA vaccine encoding gD antigen against pseudorabies virus infection.

Messenger RNA-based vaccines represent new tools with prophylactic and therapeutic potential characterized by high flexibility of application for infectious diseases. Pseudorabies virus (PRV) is one of the major viruses affecting the pig industry. PRV has serious effects in piglets, sows, and growing-fattening pigs and can lead to huge economic losses. In this study, an envelope glycoprotein D (gD) gene-based specific mRNA vaccine was generated, and a mouse model was used to investigate the protective efficacy of the vaccine. The gD mRNA vaccine and the recombinant plasmid pVAX-gD were transfected into BHK21 cells, and the antigenicity of the expressed proteins was detected by Western blot analysis. Groups of mice were vaccinated with the gD mRNA vaccine, pVAX-gD, and PBS. T cell immune responses were measured by flow cytometry or ELISA and serum neutralization tests every two weeks. The challenge with the PRV-XJ strain was performed eight weeks after the primary immunization, and the response was monitored for 15 days. The levels of specific and neutralizing antibodies in the gD mRNA vaccine group were significantly increased in 8 weeks compared to those in the control group, and cytokine levels, including that of IFN-γ/IL-2, were considerably higher than those in the control animal. Additionally, the proportion of CD4+/CD8+ cells in peripheral lymphocytes was remarkably increased. Our data demonstrate that mRNA is a promising and effective tool for the development of vaccines. The PRV-gD-based mRNA vaccine can elicit an efficient neutralizing antibody response and induce effective protection in mice in defense against PRV infection.

Deciphering the Subtype Differentiation History of SARS-CoV-2 Based on a New Breadth-First Searching Optimized Alignment Method Over a Global Data Set of 24,768 Sequences.

SARS-CoV-2 has caused a worldwide pandemic. Existing research on coronavirus mutations is based on small data sets, and multiple sequence alignment using a global-scale data set has yet to be conducted. Statistical analysis of integral mutations and global spread are necessary and could help improve primer design for nucleic acid diagnosis and vaccine development. Here, we optimized multiple sequence alignment using a conserved sequence search algorithm to align 24,768 sequences from the GISAID data set. A phylogenetic tree was constructed using the maximum likelihood (ML) method. Coronavirus subtypes were analyzed via t-SNE clustering. We performed haplotype network analysis and t-SNE clustering to analyze the coronavirus origin and spread. Overall, we identified 33 sense, 17 nonsense, 79 amino acid loss, and 4 amino acid insertion mutations in full-length open reading frames. Phylogenetic trees were successfully constructed and samples clustered into subtypes. The COVID-19 pandemic differed among countries and continents. Samples from the United States and western Europe were more diverse, and those from China and Asia mainly contained specific subtypes. Clades G/GH/GR are more likely to be the origin clades of SARS-CoV-2 compared with clades S/L/V. Conserved sequence searches can be used to segment long sequences, making large-scale multisequence alignment possible, facilitating more comprehensive gene mutation analysis. Mutation analysis of the SARS-CoV-2 can inform primer design for nucleic acid diagnosis to improve virus detection efficiency. In addition, research into the characteristics of viral spread and relationships among geographic regions can help formulate health policies and reduce the increase of imported cases.

Comparative Study of Microbiological Monitoring Results from Three Types of Sampling Methods after Gastrointestinal Endoscope Reprocessing.

OBJECTIVE: Compare the effects of three sampling methods on the microbiological monitoring results after reprocessing of gastrointestinal endoscopes, providing scientific basis for improving the monitoring quality of gastrointestinal endoscope cleaning and disinfection. METHOD: Gastrointestinal endoscopes after reprocessing were selected randomly at the gastrointestinal endoscopy center of a tertiary hospital in Shanghai from October 2018 to February 2019. The endoscopes selected were all sampled in three different methods under continuous sampling and intermittent sampling respectively. Methods used includes, the biopsy channel group (Group A), the entire channel group (Group B), and the disc brush group (Group C). Then the colony forming units (CFU/piece) were counted in the laboratory. RESULTS: A total of 12 endoscopes were sampled by using continuous sampling approach, in which the detection rate of bacteria in disc brush group (33.3%) and entire channel group (33.3%) was higher than biopsy channel group (8.3%). Among the 12 endoscopes sampled with intermittent approach, the detection rate of bacteria from high to low was the disc brush group (50%), the entire channel group (41.7%), and the biopsy channel group (8.3%). CONCLUSION: Different sampling methods will lead to the difference of microbiological culture results after reprocessing of gastrointestinal endoscope, indicating that the improved sampling method is beneficial to objectively reflect the endoscope cleaning and disinfection effect, and improve the monitoring quality of endoscope disinfection.

In vitro modulation of mercury-induced rat liver mitochondria dysfunction.

Mercury (Hg) is a toxic environmental pollutant that exerts its cytotoxic effects as cations by targeting mitochondria. In our work, we determined different mitochondrial toxicity factors using specific substrates and inhibitors following the addition of Hg2+ to the mitochondria isolated from Wistar rat liver in vitro. We found that Hg2+ induced marked changes in the mitochondrial ultrastructure accompanied by mitochondrial swelling, mitochondrial membrane potential collapse, mitochondrial membrane fluidity increase and Cytochrome c release. Additionally, the effects of Hg2+ on heat production of mitochondria were investigated using microcalorimetry; simultaneously, the effects on mitochondrial respiration were determined by Clark oxygen-electric methods. Microcalorimetry could provide detailed kinetic and thermodynamic information which demonstrated that Hg2+ had some biotoxicity effect on mitochondria. The inhibition of energy metabolic activities suggested that high concentrations of Hg2+ could induce mitochondrial ATP depletion under MPT and mitochondrial respiration inhibition. These results help us learn more about the toxicity of Hg2+ at the subcellular level.