Effect of volatile versus propofol anaesthesia on major complications and mortality after cardiac surgery: a multicentre randomised trial.

BACKGROUND: The comparative effectiveness of volatile anaesthesia and total intravenous anaesthesia (TIVA) in terms of patient outcomes after cardiac surgery remains a topic of debate. METHODS: Multicentre randomised trial in 16 tertiary hospitals in China. Adult patients undergoing elective cardiac surgery were randomised in a 1:1 ratio to receive volatile anaesthesia (sevoflurane or desflurane) or propofol-based TIVA. The primary outcome was a composite of predefined major complications during hospitalisation and mortality 30 days after surgery. RESULTS: Of the 3123 randomised patients, 3083 (98.7%; mean age 55 yr; 1419 [46.0%] women) were included in the modified intention-to-treat analysis. The composite primary outcome was met by a similar number of patients in both groups (volatile group: 517 of 1531 (33.8%) patients vs TIVA group: 515 of 1552 (33.2%) patients; relative risk 1.02 [0.92-1.12]; P=0.76; adjusted odds ratio 1.05 [0.90-1.22]; P=0.57). Secondary outcomes including 6-month and 1-yr mortality, duration of mechanical ventilation, length of ICU and hospital stay, and healthcare costs, were also similar for the two groups. CONCLUSIONS: Among adults undergoing cardiac surgery, we found no difference in the clinical effectiveness of volatile anaesthesia and propofol-based TIVA. CLINICAL TRIAL REGISTRATION: Chinese Clinical Trial Registry (ChiCTR-IOR-17013578).

Computation-Guided Rational Design of Cysteine-Less Protein Variants in Engineered hCGL.

The engineered human cystathionine-γ-lyase (hCGL) resulting in enhanced activity toward both cysteine and cystine unveils a potential robust antitumor activity. However, the presence of cysteine residues has the potential to induce oligomerization or incorrect disulfide bonding, which may decrease the bioavailability of biopharmaceuticals. Through a meticulous design process targeting the cysteine residues within engineered hCGL, a set of potential beneficial mutants were obtained by virtual screening employing Rosetta and ABACUS. Experimental measurements have revealed that most of the mutants showed increased activity toward both substrates l-Cys and CSSC. Furthermore, mutants C109V and C229D demonstrated Tm value increases of 8.2 and 1.8 °C, respectively. After an 80 min incubation at 60 °C, mutant C229D still maintained high residual activity. Unexpectedly, mutant C109V, displaying activity approximately 2-fold higher than the activity of wild type (WT) for both substrates, showed disappointing instability in plasma, which suggests that computational design still requires further consideration. Analysis of their structure and molecular dynamics (MD) simulation revealed the impact of hydrophobic interaction, hydrogen bonds, and near-attack conformation (NAC) stability on activity and stability. This study acquired information about mutants that exhibit enhanced activity or thermal resistance and serve as valuable guidance for subsequent specific cysteine modifications.

IGF2BP3-mediated enhanced stability of MYLK represses MSC adipogenesis and alleviates obesity and insulin resistance in HFD mice.

Mesenchymal stem cells (MSCs) hold immense potential as multipotent stem cells and serve as a primary source of adipocytes. The process of MSC adipogenesis plays a crucial role in maintaining systemic metabolic homeostasis and has garnered significant attention in tissue bioengineering. N6-methyladenosine (m6A), the most prevalent RNA modification, is known to regulate cell fate and disease. However, the precise involvement of m6A readers in MSC adipogenesis remains unclear. In this study, we investigated the impact of IGF2BP3, a prominent m6A reader, on MSC adipogenesis. Our findings revealed a decrease in IGF2BP3 expression during the natural adipogenic differentiation of MSCs. Furthermore, IGF2BP3 was found to repress MSC adipogenesis by augmenting the levels of MYLK, a calcium/calmodulin-dependent kinase. Mechanistically, IGF2BP3 interacted with MYLK mRNA in an m6A-dependent manner, extending its half-life and subsequently inhibiting the phosphorylation of the ERK1/2 pathway, thereby impeding the adipogenic differentiation of MSCs. Additionally, we successfully achieved the overexpression of IGF2BP3 through intraperitoneal injection of adeno-associated virus serotype Rec2, which specifically targeted adipose tissue. This intervention resulted in reduced body weight and improved insulin resistance in high-fat diet mice. Overall, our study provides novel insights into the role of IGF2BP3 in MSC adipogenesis, shedding light on adipocyte-related disorders and presenting potential targets for related biomedical applications.

Novel insights into STAT3 in renal diseases.

Signal transducer and activator of transcription 3 (STAT3) is a cell-signal transcription factor that has attracted considerable attention in recent years. The stimulation of cytokines and growth factors can result in the transcription of a wide range of genes that are crucial for several cellular biological processes involved in pro- and anti-inflammatory responses. STAT3 has attracted considerable interest as a result of a recent upsurge in study because of their role in directing the innate immune response and sustaining inflammatory pathways, which is a key feature in the pathogenesis of many diseases, including renal disorders. Several pathological conditions which may involve STAT3 include diabetic nephropathy, acute kidney injury, lupus nephritis, polycystic kidney disease, and renal cell carcinoma. STAT3 is expressed in various renal tissues under these pathological conditions. To better understand the role of STAT3 in the kidney and provide a theoretical foundation for STAT3-targeted therapy for renal disorders, this review covers the current work on the activities of STAT3 and its mechanisms in the pathophysiological processes of various types of renal diseases.

ALKBH5 facilitates CYP1B1 mRNA degradation via m6A demethylation to alleviate MSC senescence and osteoarthritis progression.

Improving health and delaying aging is the focus of medical research. Previous studies have shown that mesenchymal stem cell (MSC) senescence is closely related to organic aging and the development of aging-related diseases such as osteoarthritis (OA). m6A is a common RNA modification that plays an important role in regulating cell biological functions, and ALKBH5 is one of the key m6A demethylases. However, the role of m6A and ALKBH5 in MSC senescence is still unclear. Here, we found that the m6A level was enhanced and ALKBH5 expression was decreased in aging MSCs induced by multiple replications, H2O2 stimulation or UV irradiation. Downregulation of ALKBH5 expression facilitated MSC senescence by enhancing the stability of CYP1B1 mRNA and inducing mitochondrial dysfunction. In addition, IGF2BP1 was identified as the m6A reader restraining the degradation of m6A-modified CYP1B1 mRNA. Furthermore, Alkbh5 knockout in MSCs aggravated spontaneous OA in mice, and overexpression of Alkbh5 improved the efficacy of MSCs in OA. Overall, this study revealed a novel mechanism of m6A in MSC senescence and identified promising targets to protect against aging and OA.

RSRC2 Expression Inhibits Malignant Progression of Triple-Negative Breast Cancer by Transcriptionally Regulating SCIN Expression.

Triple-negative breast cancer (TNBC) has a shorter survival time and higher mortality rate than other molecular subtypes. RSRC2 is a newly discovered tumor suppressor gene. However, the potential functional mechanism of RSRC2 in TNBC remains unknown so far. Multiple bioinformatics databases were used. A Human Transcriptome Array 2.0 analysis, ChIP-seq analysis, ChIP-qPCR, RT-qPCR, Western blot, cell function assays in vitro and a metastatic mouse model in vivo were performed to demonstrate the role of RSRC2 in TNBC. Through the analysis of various databases, RSRC2 expression was the lowest in TNBC tissues compared to other molecular subtypes. The low expression of RSRC2 was associated with a worse prognosis for patients with breast cancer. The transcriptome array, ChIP-seq and bioinformatics analysis identified that GRHL2 and SCIN might have a close relationship with RSRC2. The functional bioinformatics enrichment analysis and functional cell experiments showed that RSRC2 was involved in cell adhesion, cell proliferation, cell migration and invasion. Furthermore, RSRC2 expression suppressed SCIN expression but not GRHL2 expression. SCIN re-expression in the RSRC2 overexpression cells or SCIN knockdown in the RSRC2 knockdown cells reversed the cellular function caused by RSRC2. Mechanistically, RSRC2 transcriptionally inhibited SCIN expression. In summary, our study reveals that RSRC2 acts as a tumor suppressor in TNBC development and progression through negatively regulating SCIN-mediated cell function, thus providing a potential target for TNBC treatment.

Enhanced Rishirilide Biosynthesis by a Rare In-Cluster Phosphopantetheinyl Transferase in Streptomyces xanthophaeus.

Phosphopantetheinyl transferases (PPTases) play important roles in activating apo-acyl carrier proteins (apo-ACPs) and apo-peptidyl carrier proteins (apo-PCPs) in both primary and secondary metabolism. PPTases catalyze the posttranslational modifications of those carrier proteins by covalent attachment of the 4'-phosphopantetheine group to a conserved serine residue. The protein-protein interactions between a PPTase and a cognate acyl or peptidyl carrier protein have important regulatory functions in microbial biosynthesis, but the molecular mechanism underlying their specific recognition remains elusive. In this study, we identified a new rishirilide biosynthetic gene cluster with a rare in-cluster PPTase from Streptomyces xanthophaeus no2. The function of this Sfp-type PPTase, SxrX, in rishirilide production was confirmed using genetic mutagenesis and biochemical characterization. We applied molecular modeling and site-directed mutagenesis to identify key residues mediating the protein-protein interaction between SxrX and its cognate ACP. In addition, six natural products were isolated from wild-type S. xanthophaeus no2 and the ΔsxrX mutant, including rishirilide A and lupinacidin A, that exhibited antimicrobial and anticancer activities, respectively. SxrX is the first Sfp-type PPTase identified from an aromatic polyketide biosynthetic gene cluster and shown to be responsible for high-level production of rishirilide derivatives. IMPORTANCE Genome mining has been a vital means for natural product drug discovery in the postgenomic era. The rishirilide-type polyketides have attracted attention due to their potent bioactivity, but the poor robustness of production hosts has limited further research and development. This study not only identifies a hyperproducer of rishirilides but also reveals a rare, in-cluster PPTase SxrX that plays an important role in boosting rishirilide biosynthesis. Experimental and computational investigations revealed new insights on the protein-protein interaction between SxrX and its cognate ACP with wide implications for understanding polyketide biosynthesis.

Gender differences in knee kinematics during weight-bearing knee flexion for patients with arthrofibrosis after anterior cruciate ligament reconstruction.

BACKGROUND: Knowledge of tibiofemoral and patellofemoral joint kinematics is important for understanding gender-related dimorphism in developing knee arthrofibrosis and advancement of related treatments. The objective of our study was to investigate gender differences existing in tibiofemoral kinematics and patellar tracking in patients with arthrofibrosis after anterior cruciate ligament (ACL) reconstruction during weight-bearing knee flexion. METHODS: The tibiofemoral and patellofemoral joint kinematics were measured in 30 patients (15 male and 15 female) with arthrofibrosis after ACL reconstruction during a lunge task, using computed tomography and dual fluoroscopic imaging system. These data were analyzed for gender differences. RESULTS: The range of tibial rotation, patellar inferior shift, tilt, and flexion were significantly decreased in the affected knee compared to the contralateral knee from 15° to 75° of knee flexion (P ≤ 0.04). Statistically significant difference was detected for medial tibial translation between male and female patients at 60° (P = 0.04) and 75° of knee flexion (P = 0.02). The tibial rotation was significantly decreased at 60° (P = 0.03) and 75° of knee flexion (P < 0.01) in females. The inferior patellar shift in females was significantly lower than that in males at 15° (P = 0.04) and 30° of knee flexion (P = 0.01). The patellar tilt was significantly lower at 60° (P = 0.02) and 75° of knee flexion (P < 0.01) in females compared to males. CONCLUSIONS: The results indicated a significant effect of gender on knee kinematics in patients with arthrofibrosis after ACL reconstruction during weight-bearing knee flexion. These gender differences in tibiofemoral kinematics and patellar tracking may warrant further investigations to determine implications for making gender-specific surgical treatments and rehabilitation programs.

Prognostic factor analysis in patients with temporomandibular disorders after reversible treatment: study protocol for a prospective cohort study in China.

INTRODUCTION: Temporomandibular disorders (TMDs) are complex multifactorial disorders. Reversible treatment has been suggested for the initial management of TMD; however, comparable therapeutic effects of different reversible intervention modalities remain controversial. Various biopsychosocial factors, which may be putative prognostic factors that influence the response to reversible treatment for TMD, have been reported to increase the risk of developing first-onset TMD. However, there is a paucity of research that aims to identify prognostic factors associated with the clinical outcomes of reversible treatment in people with TMD. The objective of this prospective cohort study is to identify prognostic factors that are associated with clinical outcomes of reversible treatment in patients with TMD and to analyse the risk factors that influence the development of chronic TMD. METHODS AND ANALYSIS: We plan to recruit 834 patients with TMD who meet the inclusion criteria. Once informed consent is obtained, baseline data, including anamnestic data, physical assessments and self-report questionnaires, will be collected from participants at their first clinic visit; subsequently, they will receive 1-4 weeks of reversible treatment. The primary treatment outcome measures will be a change in the anterior maximum mouth opening, worsening of TMD pain scores assessed using a visual analogue scale (VAS) and a reduction in characteristic pain intensity. A good outcome will be defined as an anterior maximal opening ≥35 mm and at least a 30% reduction in VAS scores 3 months after baseline. The association between candidate prognostic factors and clinical outcomes of reversible TMD treatment will be analysed. ETHICS AND DISSEMINATION: The protocol has been approved by the Ethics Committee of Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China, based on the guidelines outlined in the Declaration of Helsinki (SH9H-2019-T316-4). The results of this study will be reported in accordance with the Strengthening the Reporting of Observational Studies in Epidemiology statement. The authors intend to publish the results in a peer-reviewed journal. TRIAL REGISTRATION NUMBER: ChiCTR2000033328.

Macrophage migration inhibitory factor regulates joint capsule fibrosis by promoting TGF-ß1 production in fibroblasts.

Joint capsule fibrosis caused by excessive inflammation results in post-traumatic joint contracture (PTJC). Transforming growth factor (TGF)-ß1 plays a key role in PTJC by regulating fibroblast functions, however, cytokine-induced TGF-ß1 expression in specific cell types remains poorly characterized. Macrophage migration inhibitory factor (MIF) is a proinflammatory cytokine involved in inflammation- and fibrosis-associated pathophysiology. In this study, we investigated whether MIF can facilitate TGF-ß1 production from fibroblasts and regulate joint capsule fibrosis following PTJC. Our data demonstrated that MIF and TGF-ß1 significantly increased in fibroblasts of injured rat posterior joint capsules. Treatment the lesion sites with MIF inhibitor 4-Iodo-6-phenylpyrimidine (4-IPP) reduced TGF-ß1 production and relieved joint capsule inflammation and fibrosis. In vitro, MIF facilitated TGF-ß1 expression in primary joint capsule fibroblasts by activating mitogen-activated protein kinase (MAPK) (P38, ERK) signaling through coupling with membrane surface receptor CD74, which in turn affected fibroblast functions and promoted MIF production. Our results reveal a novel function of trauma-induced MIF in the occurrence and development of joint capsule fibrosis. Further investigation of the underlying mechanism may provide potential therapeutic targets for PTJC.

Manipulation under anaesthesia versus lysis of adhesions for arthrofibrosis of the knee: a 6-month randomized, multicentre, non-inferiority comparative effectiveness protocol.

BACKGROUND: Recent studies show similar outcomes between lysis of adhesions (LOA) and manipulation under anaesthesia (MUA) in patients with arthrofibrosis within 3 to 6 months after arthroscopic knee procedures. As MUA offers positive efficacy with less expense and more convenience, the first consideration in clinical practice shifting to MUA may save much medical cost when MUA is non-inferior to surgery. There is a lack of evidence comparing the clinical outcomes of these two surgical techniques. The purpose of our study is to determine whether MUA is non-inferior to LOA for improving clinical outcomes in patients with loss of flexion within 3 and 6 months after arthroscopic knee surgery. METHODS: This multicentre randomized controlled trial (RCT) will recruit 360 patients within 3 to 6 months after arthroscopic knee surgery with a magnetic resonance imaging (MRI)-confirmed arthrofibrosis to prove non-inferiority of MUA. Patients are randomly assigned to the LOA group (n=180) or the MUA group (n=180) in a 1:1 ratio with random sequences using the SPSS Statistics V.22. The standard surgical procedures will be performed within 2 weeks after randomization, and each patient will receive unified perioperative physical therapy (PT). The primary outcome measure is the range of knee flexion. Secondary outcome measures include physical function, activity level, general health, knee pain, and the occurrence of adverse events. Safety measures include physiological parameters (i.e., blood pressure, pulse, respiratory rate) and complications (i.e., component damage, patellar tendon tear and fracture during surgical procedures, neurological or internal medicine conditions, venous thromboembolism, bleeding and infection postoperatively). The noninferiority margin is defined as a difference of 10 degrees on the range of knee flexion between groups and was assessed with a one-sided α of .025. To investigate relative effectiveness between the groups, linear mixed-effects model will be used for continuous data, and generalised estimating equation for dichotomous data. Analyses will follow the intention-to-treat principle. DISCUSSION: Findings of our study will help clinicians determine the risk-benefit balance of MUA and LOA, two frequently used surgical procedures for patients with knee arthrofibrosis. TRIAL REGISTRATION: Chinese Clinical Trial Registry (www.chictr.org.cn); ChiCTR2000033467.

LXYL-P1-2 immobilized on magnetic nanoparticles and its potential application in paclitaxel production

BACKGROUND: LXYL-P1-2 is the first reported glycoside hydrolase that can catalyze the transformation of 7-b-xylosyl-10-deacetyltaxol (XDT) to 10-deacetyltaxol (DT) by removing the D-xylosyl group at the C7 position. Successful synthesis of paclitaxel by one-pot method combining the LXYL-P1-2 and 10- deacetylbaccatin III-10-b-O-acetyltransferase (DBAT) using XDT as a precursor, making LXYL-P1-2 a highly promising enzyme for the industrial production of paclitaxel. The aim of this study was to investigate the catalytic potential of LXYL-P1-2 stabilized on magnetic nanoparticles, the surface of which was modified by Ni2+-immobilized cross-linked Fe3O4@Histidine. RESULTS: The diameter of matrix was 20­40 nm. The Km value of the immobilized LXYL-P1-2 catalyzing XDT (0.145 mM) was lower than that of the free enzyme (0.452 mM), and the kcat/Km value of immobilized enzyme (12.952 mM s 1 ) was higher than the free form (8.622 mM s 1 ). The immobilized form maintained 50% of its original activity after 15 cycles of reuse. In addition, the stability of immobilized LXYL-P1-2, maintained 84.67% of its initial activity, improved in comparison with free form after 30 d storage at 4 C. CONCLUSIONS: This investigation not only provides an effective procedure for biocatalytic production of DT, but also gives an insight into the application of magnetic material immobilization technology.

The Impact of Education and Physical Therapy on Oral Behaviour in Patients with Temporomandibular Disorder: A Preliminary Study.

Patient education is important in the treatment of temporomandibular disorder (TMD), but little is known about its effect on oral behaviors. We aimed to determine the dominant oral behaviours in patients with TMD and assess the impact of education on such behaviours. Between July 2018 and April 2019, 54 patients diagnosed with TMD according to DC/TMD were recruited. They received physical therapy and were provided education on TMD and offered a list of recommendations for improving their oral behaviours. The patient education process usually lasted for 10-20 min. Of these patients, 48 were reexamined at the outpatient clinic, 3-9 months posttreatment. We recorded the Oral Behaviour Checklist (OBC) score, maximum painless mouth opening (mm), visual analogue scale (VAS) score for pain, and Jaw Functional Limitation Scale (JFLS) score pre- and posttreatment. Wilcoxon signed rank test and paired sample t-test were used for statistical analysis. Results showed that the most dominant oral behaviours included "putting pressure on the jaw" (59.3%); "chewing food on one side" (46.3%); "pressing, touching, or holding teeth together at times other than eating" (33.3%); and "eating between meals" (33.3%). Posttreatment, the patients reported a decrease in "chewing gum" (P = 0.002), "leaning with the hand on the jaw" (P = 0.013), "chewing food on one side" (P ≤ 0.001), and "eating between meals" (P = 0.007), but this change was not significant in subgroups with a follow-up interval of 9 months. We also observed a significant improvement in the maximum painless mouth opening (P ≤ 0.001), JFLS score (P ≤ 0.001), and VAS score (P ≤ 0.001) for pain, posttreatment. In conclusion, patient education can facilitate management of oral behaviours and should be targeted towards specific oral behaviours.

Regioselective C-H Amidation of (Alkyl)arenes by Iron(II) Catalysis.

A nondirected amidation reaction of aromatic C-H bond was developed under iron(II) catalysis, using sulfonyl azides as the nitrogen source. The reaction displayed a broad substrate scope and good regioselectivities in the aspects of aromatic ring vs alkyl chain and different aromatic position of (alkyl)arenes. This method provided a new protocol for the synthesis of some aromatic amines, which were hard to achieve in a previous report.

Insights into the Phosphoryl Transfer Mechanism of Human Ubiquitous Mitochondrial Creatine Kinase.

Human ubiquitous mitochondrial creatine kinase (uMtCK) is responsible for the regulation of cellular energy metabolism. To investigate the phosphoryl-transfer mechanism catalyzed by human uMtCK, in this work, molecular dynamic simulations of uMtCK∙ATP-Mg2+∙creatine complex and quantum mechanism calculations were performed to make clear the puzzle. The theoretical studies hereof revealed that human uMtCK utilizes a two-step dissociative mechanism, in which the E227 residue of uMtCK acts as the catalytic base to accept the creatine guanidinium proton. This catalytic role of E227 was further confirmed by our assay on the phosphatase activity. Moreover, the roles of active site residues in phosphoryl transfer reaction were also identified by site directed mutagenesis. This study reveals the structural basis of biochemical activity of uMtCK and gets insights into its phosphoryl transfer mechanism.

The whole-cell immobilization of D-hydantoinase-engineered Escherichia coli for D-CpHPG biosynthesis

Background: D-Hydroxyphenylglycine is considered to be an important chiral molecular building-block of antibiotic reagents such as pesticides, and β-lactam antibiotics. The process of its production is catalyzed by D-hydantoinase and D-carbamoylase in a two-step enzyme reaction. How to enhance the catalytic potential of the two enzymes is valuable for industrial application. In this investigation, an Escherichia coli strain genetically engineered with D-hydantoinase was immobilized by calcium alginate with certain adjuncts to evaluate the optimal condition for the biosynthesis of D-carbamoyl-p-hydroxyphenylglycine (D-CpHPG), the compound further be converted to D-hydroxyphenylglycine (D-HPG) by carbamoylase. Results: The optimal medium to produce D-CpHPG by whole-cell immobilization was a modified Luria-Bertani (LB) added with 3.0% (W/V) alginate, 1.5% (W/V) diatomite, 0.05% (W/V) CaCl2 and 1.00 mM MnCl2.The optimized diameter of immobilized beads for the whole-cell biosynthesis here was 2.60 mm. The maximized production rates of D-CpHPG were up to 76%, and the immobilized beads could be reused for 12 batches. Conclusions: This investigation not only provides an effective procedure for biological production of D-CpHPG, but gives an insight into the whole-cell immobilization technology.

The identification of goat peroxiredoxin-5 and the evaluation and enhancement of its stability by nanoparticle formation.

An anticancer bioactive peptide (ACBP), goat peroxiredoxin-5 (gPRDX5), was identified from goat-spleen extract after immunizing the goat with gastric cancer-cell lysate. Its amino acid sequence was determined by employing 2D nano-LC-ESI-LTQ-Orbitrap MS/MS combined with Mascot database search in the goat subset of the Uniprot database. The recombinant gPRDX5 protein was acquired by heterogeneous expression in Escherichia coli. Subsequently, the anti-cancer bioactivity of the peptide was measured by several kinds of tumor cells. The results indicated that the gPRDX5 was a good anti-cancer candidate, especially for killing B16 cells. However, the peptide was found to be unstable without modification with pharmaceutical excipients, which would be a hurdle for future medicinal application. In order to overcome this problem and find an effective way to evaluate the gPRDX5, nanoparticle formation, which has been widely used in drug delivery because of its steadiness in application, less side-effects and enhancement of drug accumulation in target issues, was used here to address the issues. In this work, the gPRDX5 was dispersed into nanoparticles before delivered to B16 cells. By the nanotechnological method, the gPRDX5 was stabilized by a fast and accurate procedure, which suggests a promising way for screening the peptide for further possible medicinal applications.