Nicotinamide N-Methyl Transferase as a Predictive Marker of Tubular Fibrosis in CKD.

Purpose: Chronic kidney disease (CKD) progression is complex. There are not standardized methods for predicting the prognosis of CKD. Nicotinamide N-methyltransferase (NNMT) has been shown to be associated with renal fibrosis. This study aimed to validate NNMT as a prognostic biomarker of progressive CKD. Patients and Methods: We explored the relationship between NNMT expression and CKD-related outcome variables using the NephroseqV5 and GEO databases. Additionally, a validation set of 37 CKD patients was enrolled to measure the correlation between NNMT expression levels and CKD outcomes. Furthermore, single-cell RNA sequencing data and the Human Protein Atlas were reanalyzed to investigate the expression specificity of NNMT in the kidney. Finally, to detect the status of NNMT expression with tubular fibrosis in vivo, we constructed a unilateral ureteral obstruction (UUO) mouse treated with an NNMT inhibitor. Results: Analyzing the datasets showed that NNMT was expressed mainly in proximal tubule compartments. And patients with high NNMT expression levels had a significantly lower overall survival rate compared to those with low NNMT expression levels (P = 0.013). NNMT was independent of prognosis factors in the multivariate Cox regression model, and the AUCs for CKD progression at 1, 3, and 5 years were 0.849, 0.775, and 0.877, respectively. Pathway enrichment analysis indicated that NNMT regulates the biological processes of tubulointerstitial fibrosis (TIF). In the validation group, NNMT levels were significantly higher in the CKD group combined with interstitial fibrosis. In vivo, NNMT was a high expression in the UUO group, peaking at postoperative day 21. Treatment with an NNMT inhibitor improved renal tubular interstitial fibrosis, and expression levels of FN, α-SMA, VIM, and TGF-ß1 were decreased compared with UUO (P < 0.05). Conclusion: NNMT was expressed mainly in tubular renal compartments, and associated with CKD prognosis. It holds potential as a diagnostic biomarker for tubular fibrosis in CKD.

CCNB1 is involved in bladder cancer pathogenesis and silencing CCNB1 decelerates tumor growth and improves prognosis of bladder cancer.

In search of an effective therapeutic target for bladder urothelial carcinoma (BLCA), the present study aimed to investigate the expression of cyclin B1 (CCNB1) and its putative mechanism in BLCA. BLCA sequencing data from Gene Expression Omnibus and The Cancer Genome Atlas were used to analyze expression of CCNB1 mRNA and high CCNB1 expression had a poorer prognosis compared with those with low expression. Immunohistochemistry (IHC) samples collected from the Human Protein Atlas database were analyzed for CCNB1 protein expression. Short hairpin (sh) CCNB1-transfected BLCA T24 and 5637 cells were used to investigate the effects of CCNB1 and inhibit the proliferation, migration and invasion of BLCA cells, affect the cell cycle distribution and promote apoptosis of 5637 cells. A sh-CCNB1 BLCA chicken embryo chorioallantoic membrane (CAM) transplantation model was established to observe the impacts of sh-CCNB1 on the tumorigenesis of BLCA in vivo. Analysis of sequencing data showed that CCNB1 mRNA was significantly elevated in tumor and BLCA compared with normal tissues [standardized mean difference (SMD)=1.21; 95% CI: 0.26-2.15; I²=95.9%]. IHC indicated that CCNB1 protein was localized in the nucleus and cytoplasm and was significantly increased in BLCA tumor tissues. The in vitro tests demonstrated that proliferation of T24 and 5637 cells transfected with sh-CCNB1 was significantly inhibited and cell migration and invasion ability were significantly decreased. sh-CCNB1 decreased the percentage of T24 cells in G0/G1, 5637 cells in the G0/G1 phase and S phase and increased percentage of 5637 cells in the G2/M phase and increased early apoptosis of 5637 cells. The in vivo experiments demonstrated that the mass of transplanted tumors was significantly decreased compared with the control group following silencing of CCNB1. The present results suggested that CCNB1 was involve in the development and prognosis of BLCA and silencing of CCNB1 may be a promising targeted therapy for BLCA.

The Regulation Effect of Team Leadership and Cohesion on Athletes' Sports Grade / El efecto de regulación del liderazgo y la cohesión del equipo en el grado deportivo de los atletas

Team cohesion is a crucial element in bringing the group together. Team cohesion is the engine that drives the progression of a sports team, and it plays a crucial part in boosting the team's confidence and overall performance. This study aims to investigate the connection between athlete leadership and team cohesion and the potential mediating and regulating effects between emotional infection and sports grade. The Athlete Leadership Questionnaire, Group Environment Questionnaire, and Emotional Infection Scale were administered to 316 athletes, and the data were analyzed using SPSS. The Bootstrap approach was used to determine whether emotional infection significantly mediated the link between athlete conduct and team cohesion. In the main effect, athlete leadership can positively and significantly predict team cohesion (β=0.531, p<0.01), and the interaction item between athlete's leadership and sports level also reached a significant level (β=0.355, p<0.01). Age and sex did not pass the F test (F=1.148, p>0.05), indicating that they will not affect group cohesion. This study found that team cohesiveness and external leaders had the most impact on social cohesion, but social leaders had the greatest impact on task cohesion. In addition, the study found that emotional infection serves as a partial mediator between the leadership of athletes and their numerous aspects and team cohesion.(AU)

Coaches’ leadership behavior, motivational atmosphere and athletes’ motivational internalization: the perspective of self-determination theory / Comportamiento de liderazgo de los entrenadores, atmósfera motivacional e internalización motivacional de los atletas: la perspectiva de la teoría de la autodeterminación

Objective: The objective of this study is to examine the leadership behaviour of coaches and the relationship between the mastery motivational atmosphere and the respective sub-dimensions of basic psychological needs. This is done with a view to better solve the path selection and optimization problem noted in coaches' leadership behaviour’s effectiveness based on the self-determination theory.Methods: This research focuses on volleyball players and uses the maximum likelihood method to estimate the hypothetical model by ccarrying out the path coefficient significance test.Results: In the incentive atmosphere of cooperative learning, athletes will continue to find problems, correct mistakes, and improve their understanding of training and competition. Therefore, it is reasonable to assume that the training-related guidance of coaches plays a key role in the construction of an incentive atmosphere of cooperative learning.Conclusion: The coaches’ positive behaviours, tolerance of failure, and encouragement of attempts can help build a task-oriented motivational atmosphere, and through the transfer of athletes to realize the internalization of external incentives, athletes show a positive behaviour orientation.(AU)

Study on the influencing factors of removal of NOX from cement kiln flue gas by sewage sludge as a denitration agent.

Experimental study on the influencing factors of using sewage sludge as a denitration agent for cement industry was carried out on a self-made laboratory-scale fluid-bed reactor. Results indicate that sludge combustion at 900 °C shows an ideal NOX (the sum of NO and NO2) removal activity under simulated working conditions of cement precalciner. The optimal removal efficiency of NOX can reach 70.36 ± 3.59% in the presence of cement raw meal (CRM) at a sludge particle size range of 0.18-0.25 mm and the sludge dosage of 0.75 g/min. Besides, the NOX removal efficiency increases to 76.94 ± 5.02% in the absence of CRM, indicating that cement raw meal inhibits the NOX removal. This phenomenon may be attributed to the fact that CRM has promotion effect on NH3 produced and obvious inhibitory effect on CO produced; while NH3 and CO play a leading role in NOX reduction, the combined effect leads to the decrease of NOX removal. Moreover, the relationship between the composition of CRM on the inhibition of NOX removal is MgO < CaCO3 < CRM < Al2O3

Long-term effect of ropivacaine nanoparticles for sciatic nerve block on postoperative pain in rats.

PURPOSE: The analgesic effect of ropivacaine (Rop) for nerve block lasts only ~3-6 hours for single use. The aim of this study was to develop long-acting regional anesthetic Rop nanoparticles and investigate the effects of sciatic nerve block on postoperative pain in rats. MATERIALS AND METHODS: Rop nanoparticles were developed using polyethylene glycol-co-polylactic acid (PELA). One hundred and twenty adult male Wistar rats were randomly divided into four groups (n=30, each): Con (control group; 0.9% saline, 200 µL), PELA (PELA group; 10 mg), Rop (Rop group; 0.5%, 200 µL), and Rop-PELA (Rop-PELA group; 10%, 10 mg). Another 12 rats were used for the detection of Rop concentration in plasma. The mechanical withdrawal threshold and thermal withdrawal latency were measured at 2 hours, 4 hours, 8 hours, 1 day, 2 days, 3 days, 5 days, and 7 days after incision. The expression of c-FOS was determined by immunohistochemistry at 2 hours, 8 hours, 48 hours, and 7 days. Nerve and organ toxicities were also evaluated at 7 days. RESULTS: The duration of Rop absorption in the plasma of the Rop-PELA group was longer (>8 hours) than that of the Rop group (4 hours). Mechanical withdrawal threshold and thermal withdrawal latency in the Rop-PELA group were higher than that in other groups (4 hours-3 days). c-FOS expression in the Rop-PELA group was lower than that in the control group at 2 hours, 8 hours, and 48 hours and lower than that in the Rop group at 8 hours and 48 hours after paw incision. Slight foreign body reactions were observed surrounding the sciatic nerve at 7 days. No obvious pathophysiological change was found in the major organs after Rop-PELA administration at 7 days. CONCLUSION: Rop-PELA provides an effective analgesia for nerve block over 3 days after single administration, and the analgesic mechanism might be mediated by the regulation of spinal c-FOS expression. However, its potential long-term tissue toxicity needs to be further investigated.

[Detection of binding of antibodies with recombinant house dust mite allergen (rDer p2) by surface plasmon resonance technique].

In the present paper, the authors aimed to detect the binding of monoclonal antibody and polyclonal antibodies in sera of patients with recombinant house dust mite allergen (rDer p2) by surface plasmon resonance (SPR) technique. This technique is superior to other methods, like enzyme-linked immunosorbent assays (ELISAs) in studying the interaction between biomolecules because no labeling and sample separation are needed. The allergen rDer p2 was immobilized on carboxymethyldextran-modified sensor chip surface by amine coupling. Surface plasmon resonance measurements of monoclonal antibody and polyclonal antibodies in patients' sera revealed that their bindings diverge widely; the binding of patients' sera was remarkably lower than that of monoclonal antibody. At the same time, the binding of patients' sera with rDer p2 varied among patients allergic to dust mite. This study could provide an easy, fast and real-time way for clinical allergic diseases diagnosis.

Impedance sensing of allergen-antibody interaction on glassy carbon electrode modified by gold electrodeposition.

The interactions between the recombinant dust mite allergen Der f2 and murine monoclonal antibody were monitored by electrochemical impedance spectroscopy (EIS). Allergen Der f2 were immobilized through the nanogold formed by electrodeposition of gold on planar glassy carbon electrode. A 30-s gold electrodeposition provided a desirable substrate for the immobilization of allergen. Electrochemical deposition of gold on a glassy carbon electrode showed significant improvement in allergen immobilization. The impedance measurements were based on the charge-transfer kinetics of the [Fe(CN)(6)](3-/4-) redox pair. The interactions between allergen and antibody occurred on electrode surface altered the interfacial electron transfer resistance, R(CT), by preventing the redox species approaching the electrode. The results showed that R(CT) increased with increasing concentration of monoclonal antibodies.

[Study on structures of blattela germanica allergen, Bla g 2 expressed by eukaryotic and prokaryotic vectors using fluorescence and CD spectra].

The recombinant allergen, Bla g 2, was expressed by prokaryotic vector E. coli and eukaryotic vector P. Pastoris. The different structures and configurations of the Bla g 2 from E. coli and P. Pastoris were studied by fluorescence and circular dichroism. The secondary structures of Bla g 2 in solution, and the composition besides the type of its tertiary structure were proposed. These studies help understand the differences between prokaryotic and eukaryotic expression systems, reveal the relationship between the structure and the function of Bla g 2, and improve the production of this significant allergen.

Application of electrochemical impedance spectroscopy for monitoring allergen-antibody reactions using gold nanoparticle-based biomolecular immobilization method.

Gold nanoparticles were used to enhance the immobilization amount and retain the immunoactivity of recombinant dust mite allergen Der f2 immobilized on a glassy carbon electrode (GCE). The interaction between allergen and antibody was studied by electrochemical impedance spectroscopy (EIS). Self-assembled Au colloid layer (ø=16nm) deposited on (3-mercaptopropyl)trimethoxysilane (MPTS)-modified GCE offered a basis to control the immobilization of allergen Der f2. The impedance measurements were based on the charge transfer kinetics of the [Fe(CN)(6)](3-/4-) redox pair, compared with bare GCE, the immobilization of allergen Der f2 and the allergen-antibody interaction that occurred on the electrode surface altered the interfacial electron transfer resistance and thereby slowed down the charge transfer kinetics by reducing the active area of the electrode or by preventing the redox species in electrolyte solution from approaching the electrode. The interactions of allergen with various concentrations of monoclonal antibody were also monitored through the change of impedance response. The results showed that the electron transfer resistance increased with increasing concentrations of monoclonal antibody.

Buildup of gold nanoparticle multilayer thin films based on the covalent-bonding interaction between boronic acids and polyols.

A novel method for the fabrication of gold nanoparticle multilayer films based on the covalent-bonding interaction between boronic acid and polyols, poly(vinyl alcohol) (PVA), was developed. The multilayer buildup was monitored by UV-vis absorbance spectroscopy, which showed a linear increase of the film absorbance with the number of adsorbed Au layers and indicated the stepwise and uniform assembling process. The atomic force microscopy (AFM) image showed that a compact gold multilayer thin film was successfully assembled. The residual boronic acid group on the surface of thin film could incorporate glycosylated-protein horseradish peroxidase (HRP), and good catalytic activity for H2O2 could be observed.

[Study of fluorescence spectra of blattela germanica allergen, derenatured and renatured Bla g 2].

Recombinant proteins extracted from inclusion body remain in denaturation status. Renaturation in vitro after initial purification is a key step of downstream processing. A common method of renaturation of recombinant proteins is the dilution method. With Bla g 2 as a model protein, the conformational changes of denatured and renatured Bla g 2 were investigated by applying fluorescence spectra. The effects of different urea concentrations, different SDS concentrations and different pH on the fluorescence intensity of renatured protein were also investigated. The reasons for these were studied with the knowledge of molecular structure.

Morphology study of gold-chitosan nanocomposites.

Gold nanoparticles were prepared in the presence of chitosan via reduction of HAuCl4 with sodium borohydride. The gold-chitosan nanocomposite was formed by adsorbing chitosan molecules onto the gold nanoparticle surfaces. The resulting gold nanoparticles were characterized by transmission electron microscopy and UV-vis spectroscopy. Morphology of gold-chitosan nanocomposite films was investigated by polarized optical microscopy. The morphology of chitosan crystal cast from the prepared nanocomposite was much different from that cast from chitosan solution due to the possible nucleation of gold nanoparticles. A branched-like structure or a cross-linked needle-like structure could be formed in nanocomposite films with different casting volumes.

Synthesis of chitosan-stabilized gold nanoparticles in the absence/presence of tripolyphosphate.

Gold nanoparticles were prepared by reducing gold salt with a polysaccharide, chitosan, in the absence/presence of tripolyphosphate (TPP). Here, chitosan acted as a reducing/stabilizing agent. The obtained gold nanoparticles were characterized with UV--vis spectroscopy and transmission electron microscopy. The results indicated that the shape and size distribution of gold nanoparticles changed with the molecular weight and concentration of chitosan. More interestingly, the gelation of chitosan upon contacting with polyanion (TPP) can also affect the shape and size distribution of gold nanoparticles. By adding TPP to chitosan solution before the reduction of gold salt, gold nanoparticles have a bimodal size distribution, and at the same time, polygonal gold particles were obtained in addition to spherical gold nanoparticles.

Preparation and characterization of metal-chitosan nanocomposites.

Various metal-chitosan nanocomposites were synthesized, including silver (Ag), gold (Au), platinum (Pt), and palladium (Pd) in aqueous solutions. Metal nanoparticles were formed by reduction of corresponding metal salts with NaBH4 in the presence of chitosan. And chitosan molecules adsorbing onto the surface of as-prepared metal nanoparticles formed the corresponding metal-chitosan nanocomposites. Transmission electron microscopy (TEM) images and UV-vis spectra of the nanocomposites revealed the presence of metal nanoparticles. Comparison of all the resulting particles size, it shows that silver nanoparticles are much larger than others (Au, Pt and Pd). In addition, the difference in particles size leads to develop different morphologies in the films cast from prepared metal-chitosan nanocomposites. Polarized optical microscopy (POM) images show a batonet-like structure for Ag-chitosan nanocomposites film, while for the films cast from other metal (Au, Pt, and Pd)-chitosan nanocomposites, some branched-like structures with a few differences among them were observed under POM observation.

Synthesis of polysaccharide-stabilized gold and silver nanoparticles: a green method.

A simple, green method was developed for the synthesis of gold and silver nanoparticles by using polysaccharides as reducing/stabilizing agents. The obtained positively charged chitosan-stabilized gold nanoparticles and negatively charged heparin-stabilized silver nanoparticles were characterized with UV-vis spectroscopy and transmission electron microscopy. The results illustrated the formation of gold and silver nanoparticles inside the nanoscopic polysaccharide templates. Moreover, the morphology and size distribution of prepared gold and silver nanoparticles varied with the concentration of both the polysaccharides and the precursor metal salts.