A machine learning approach for dynamical modelling of Al distributions in zeolites via23Na/27Al solid-state NMR.

One of the main limitations in supporting experimental characterization of Al siting/pairing via modelling is the high computational cost of ab initio calculations. For this reason, most works rely on static or very short dynamical simulations, considering limited Al pairing/siting combinations. As a result, comparison with experiment suffers from a large degree of uncertainty. To alleviate this limitation we have developed neural network potentials (NNPs) which can dynamically sample across broad configurational and chemical spaces of sodium-form aluminosilicate zeolites, preserving the level of accuracy of the ab initio (dispersion-corrected metaGGA) training set. By exploring a wide range of Al/Na arrangements and a combination of experimentally relevant Si/Al ratios, we found that the 23Na NMR spectra of dehydrated high-silica CHA zeolite offer an opportunity to assess the distribution and pairing of Al atoms. We observed that the 23Na chemical shift is sensitive not only to the location of sodium in 6- and 8MRs, but also to the Al-Sin-Al sequence length. Furthermore, neglect of thermal and dynamical contributions was found to lead to errors of several ppm, and has a profound influence on the shape of the spectra and the dipolar coupling constants, thus necessitating the long-term dynamical simulations made feasible by NNPs. Finally, we obtained a predictive regression model for the 23Na chemical shift in CHA (Si/Al = 35, 17, 11) that circumvents the need for expensive NMR density functional calculations and can be easily extended to other zeolite frameworks. By combining NNPs and regression methods, we can expedite the simulations of NMR properties and capture the effect of dynamics on the spectra, which is often overlooked in computational studies despite its clear manifestation in experimental setups.

Effects of sodium-glucose cotransporter 2 inhibitors on bone metabolism in patients with type 2 diabetes mellitus: a systematic review and meta-analysis.

BACKGROUND: Sodium glucose cotransporter 2 (SGLT2) inhibitors are widely used in type 2 diabetes mellitus (T2DM) therapy. The impact of SGLT2 inhibitors on bone metabolism has been widely taken into consideration. But there are controversial results in the study on the effect of SGLT2 inhibitors on bone metabolism in patients with T2DM. Therefore, we aimed to examine whether and to what extent SGLT2 inhibitors affect bone metabolism in patients with T2DM. METHODS: A literature search of randomized controlled trials (RCTs) was conducted through PubMed, Web of Science, Embase, Cochrane databases, and Scopus from inception until 15 April 2023. Eligible RCTs compared the effects of SGLT2 inhibitors versus placebo on bone mineral density and bone metabolism in patients with T2DM. To evaluate the differences between groups, a meta-analysis was conducted using the random effects inverse-variance model by utilizing standardized mean differences (SMD). RESULTS: Through screening, 25 articles were finally included, covering 22,828 patients. The results showed that, compared with placebo, SGLT2 inhibitors significantly increased parathyroid hormone (PTH, SMD = 0.13; 95%CI: 0.06, 0.20), and cross-linked C-terminal telopeptides of type I collagen (CTX, SMD = 0.11; 95%CI: 0.01, 0.21) in patients with T2DM, decreased serum alkaline phosphatase levels (ALP, SMD = -0.06; 95%CI: -0.10, -0.03), and had no significant effect on bone mineral density (BMD), procollagen type 1 N-terminal propeptide (P1NP), 25-hydroxy vitamin D, tartrate resistant acid phosphatase-5b (TRACP-5b) and osteocalcin. CONCLUSIONS: SGLT2 inhibitors may negatively affect bone metabolism by increasing serum PTH, CTX, and decreasing serum ALP. This conclusion needs to be verified by more studies due to the limited number and quality of included studies. SYSTEMATIC REVIEW REGISTRATION: PROSPERO, identifier CRD42023410701.

Caffeic Acid O-Methyltransferase Gene Family in Mango (Mangifera indica L.) with Transcriptional Analysis under Biotic and Abiotic Stresses and the Role of MiCOMT1 in Salt Tolerance.

Caffeic acid O-methyltransferase (COMT) participates in various physiological activities in plants, such as positive responses to abiotic stresses and the signal transduction of phytohormones. In this study, 18 COMT genes were identified in the chromosome-level reference genome of mango, named MiCOMTs. A phylogenetic tree containing nine groups (I-IX) was constructed based on the amino acid sequences of the 71 COMT proteins from seven species. The phylogenetic tree indicated that the members of the MiCOMTs could be divided into four groups. Quantitative real-time PCR showed that all MiCOMT genes have particularly high expression levels during flowering. The expression levels of MiCOMTs were different under abiotic and biotic stresses, including salt and stimulated drought stresses, ABA and SA treatment, as well as Xanthomonas campestris pv. mangiferaeindicae and Colletotrichum gloeosporioides infection, respectively. Among them, the expression level of MiCOMT1 was significantly up-regulated at 6-72 h after salt and stimulated drought stresses. The results of gene function analysis via the transient overexpression of the MiCOMT1 gene in Nicotiana benthamiana showed that the MiCOMT1 gene can promote the accumulation of ABA and MeJA, and improve the salt tolerance of mango. These results are beneficial to future researchers aiming to understand the biological functions and molecular mechanisms of MiCOMT genes.

Improving yield-related traits by editing the promoter of the heading date gene Ehd1 in rice.

KEY MESSAGE: We developed an efficient promoter editing method to create different weak Ehd1 alleles in elite japonica rice variety ZJ8 with slightly delayed heading and improved yield for use in breeding. Heading date is an important agronomic trait of rice (Oryza sativa) that determines the planting areas and cultivation seasons of different varieties, thus affecting final yield. Early heading date 1 (Ehd1) is a major rice integrator gene in the regulatory network of heading date whose expression level is negatively correlated with heading date and grain yield. Some elite japonica varieties such as Zhongjia 8 (ZJ8) show very early heading with poor agronomic traits when planted in South China. This problem can be addressed by downregulating the expression of Ehd1. In this study, we analyzed the cis-regulatory elements in the Ehd1 promoter region. We then used CRISPR/Cas9-mediated editing to modify the Ehd1 promoter at multiple target sites in ZJ8. We rapidly identified homozygous allelic mutations in the T2 generation via long-read sequencing. We obtained several Ehd1 promoter mutants with different degrees of lower Ehd1 expression, delayed heading date, and improved yield-related traits. We developed an efficient promoter editing method to create different weak Ehd1 alleles for breeding selection. Using this method, a series of heading date materials from elite varieties can be created to expand the planting area of rice and improve grain yields.

The efficacy and safety of patent Foramen Ovale Closure for Refractory Epilepsy (PFOC-RE): a prospectively randomized control trial of an innovative surgical therapy for refractory epilepsy patients with PFO of high-grade right-to-left shunt.

BACKGROUND: A significant proportion of patients with epilepsy have an unknown etiology and lack effective targeted therapeutic drugs. Patent Foramen Ovale (PFO) induces hypoxia and microembolism, leading to cerebral neurological dysfunction and increased epilepsy risk. This study aims to assess the efficacy and safety of PFO closure for relieving epileptic seizures in patients with refractory epilepsy associated with PFO. METHODS/DESIGN: Recruitment takes place at the West China Hospital of Sichuan University, China, for an open-label, randomized controlled clinical trial. The trial will include 110 patients with refractory epilepsy and PFO. Disease diagnoses will conform to the diagnostic criteria of the International League Against Epilepsy (ILAE) for refractory epilepsy and the American Society of Echocardiography (ASE) for PFO. Refractory epilepsy and high-grade right-to-left shunt (RLS) of the PFO will be further diagnosed using 24-hour video electroencephalogram and transthoracic echocardiography with contrast injection, respectively. Eligible participants require a secondary or higher volume of RLS. TRIAL REGISTRATION: Chinese Clinical Trial Registry (ChiCTR2200065681). Registered on November 11, 2022.

A Cu-Based Metal-Organic Framework Cu-Cip with Cuproptosis for Cancer Therapy and Inhibition of Cancer Cell Migration.

Microflora within cancer cells plays a pivotal role in promoting metastasis of cancer. However, contemporary anticancer research often overlooks the potential benefits of combining anticancer and antibacterial agents. Consequently, a metal-organic framework Cu-Cip with cuproptosis and antibacterial properties was synthesized for cancer therapy. To enhance the anticancer effect of the material, Mn2+ was loaded into Cu-Cip, yielding Mn@Cu-Cip. The fabricated material was characterized using single-crystal X-ray diffraction, PXRD, and FT-IR. By interacting with overexpressed H2O2 to produce ROS and accumulating Cu ions in cancer cells, MOFs exhibited excellent anticancer performance. Moreover, the material displayed the function of damaging Staphylococcus aureus and Escherichia coli, revealing the admirable antibacterial properties of the material. In addition, the antibacterial ability could inhibit tumor cell migration. The Cu-based MOF revealed promising applications in the field of tumor treatment.

Barriers and facilitators of implementing the practice programme for upright positions in the second stage of labour: A mixed-method study.

OBJECTIVE: To explore the possible barriers and facilitators to implementing the Upright Positions in the Second Stage of Labour (UPSSL) programme in Chinese healthcare settings. DESIGN: A mixed-method convergent design with the guidance of Consolidated Framework for Implementation Research (CFIR). METHODS: An online survey study and semi-structured interviews were conducted between March and May 2023. Healthcare professionals were recruited from four hospitals in Shijiazhuang, China. One hundred and thirty-one participants completed the survey study, and 23 of them were interviewed individually. Descriptive statistics evaluated the possible barriers and facilitators of implementing the UPSSL programme within the CFIR framework quantitatively. Guided by the CFIR framework, qualitative data were analysed using directed content analysis to summarize healthcare professionals' perspectives on barriers and facilitators of the UPSSL programme. RESULTS: Multiple intersectional barriers and facilitators were identified from the survey and semi-interviews. Healthcare professionals believed that the UPSSL programme has a scientific evidence base, systematic contents, and possible benefits for women. However, various barriers existed at individual, system, and organizational levels. Major barriers included healthcare professionals and women's safety concerns towards the use of upright positions during childbirth, the healthcare professionals' unfamiliarity with assisting an upright position birth, poor adaptability of the programme protocol, inadequate facilities and staffing, and a lack of readiness to change in the clinical setting. CONCLUSIONS: To facilitate the implementation of the UPSSL programme in China, tailored antenatal education on upright positions, especially addressing safety-related issues, should be provided to pregnant women, their families, or peers to enhance their understanding of and familiarity with such positions. Healthcare professionals should also be offered adequate training opportunities and necessary facilities. Furthermore, national-level policy changes might be required to address midwifery workforce shortages. Additionally, further research is warranted to select, adapt, and test effective implementation strategies for programme adoption. IMPLICATIONS FOR THE PROFESSION AND/OR PATIENT CARE: What problem did the study address? The adoption of upright positions during the second stage of labour could promote better maternal and neonatal outcomes and a positive childbirth experience. However, the adoption of upright positions during the second stage of labour is suboptimal in healthcare settings in China. Barriers and facilitators of implementing upright positions during childbirth are unclear. What were the main findings? A range of barriers and facilitators within the CFIR framework to promote upright positions during childbirth from healthcare professionals' perspectives were identified, and the major barriers included safety concerns towards and unfamiliarity with an upright position birth, inadequate facilities and staffing, and a lack of readiness to change in the clinical setting. Where and on whom will the research have an impact? This study will enable a better understanding of the barriers and facilitators to promoting upright positions in the second stage of labour in China. The smooth and effective implementation of the UPSSL programme could help to promote better maternal and neonatal outcomes and improve women's childbirth experiences. REPORTING METHOD: The reporting of this study followed the Consolidated Criteria for Reporting Qualitative Research (COREQ) and Good Reporting of A Mixed Methods Study (GRAMMS) guidelines. PATIENT OR PUBLIC CONTRIBUTION: In this study, healthcare professionals were involved in refining the topic guides and survey questions. Additionally, findings from the interviews were returned to them for comments and corrections.

Modern Image-Guided Surgery: A Narrative Review of Medical Image Processing and Visualization.

Medical image analysis forms the basis of image-guided surgery (IGS) and many of its fundamental tasks. Driven by the growing number of medical imaging modalities, the research community of medical imaging has developed methods and achieved functionality breakthroughs. However, with the overwhelming pool of information in the literature, it has become increasingly challenging for researchers to extract context-relevant information for specific applications, especially when many widely used methods exist in a variety of versions optimized for their respective application domains. By being further equipped with sophisticated three-dimensional (3D) medical image visualization and digital reality technology, medical experts could enhance their performance capabilities in IGS by multiple folds. The goal of this narrative review is to organize the key components of IGS in the aspects of medical image processing and visualization with a new perspective and insights. The literature search was conducted using mainstream academic search engines with a combination of keywords relevant to the field up until mid-2022. This survey systemically summarizes the basic, mainstream, and state-of-the-art medical image processing methods as well as how visualization technology like augmented/mixed/virtual reality (AR/MR/VR) are enhancing performance in IGS. Further, we hope that this survey will shed some light on the future of IGS in the face of challenges and opportunities for the research directions of medical image processing and visualization.

Dynamical Equilibrium between Brønsted and Lewis Sites in Zeolites: Framework-Associated Octahedral Aluminum.

While the structures of Brønsted acid sites (BAS) in zeolites are well understood, those of Lewis acid sites (LAS) remain an active area of investigation. Under hydrated conditions, the reversible formation of framework-associated octahedral aluminum has been observed in zeolites in the acidic form. However, the structure and formation mechanisms are currently unknown. In this work, combined experimental 27 Al NMR spectroscopy and computational data reveal for the first time the details of the zeolite framework-associated octahedral aluminium. The octahedral LAS site becomes kinetically allowed and thermodynamically stable under wet conditions in the presence of multiple nearby BAS sites. The critical condition for the existence of such octahedral LAS appears to be the availability of three protons: at lower proton concentration, either by increasing the Si/Al or by ion-exchange to non-acidic form, the tetrahedral BAS becomes thermodynamically more stable. This work resolves the question about the nature and reversibility of framework-associated octahedral aluminium in zeolites.

Evaluation of droplet digital PCR rapid detection method and precise diagnosis and treatment for suspected sepsis (PROGRESS): a study protocol for a multi-center pragmatic randomized controlled trial.

BACKGROUND: Sepsis is still a major public health concern and a medical emergency due to its high morbidity and mortality. Accurate and timely etiology diagnosis is crucial for sepsis management. As an emerging rapid and sensitive pathogen detection tool, digital droplet PCR (ddPCR) has shown promising potential in rapid identification of pathogens and antimicrobial resistance genes. However, the diagnostic value and clinical impact of ddPCR tests remains to be studied in patients with suspected sepsis. PROGRESS trial is aimed to evaluate the clinical effectiveness of a novel ddPCR assay compared with standard practice. METHODS: PROGRESS is a multicenter, open-label, pragmatic randomized controlled trial (pRCT) set in ten hospitals, including departments of infectious disease and intensive care units. In this study, a total of 2292 patients with suspected sepsis will be randomly assigned to two arms: the ddPCR group and the control group with a ratio of 3:1. The primary outcome is the diagnostic efficacy, that is, the sensitivity and specificity of the ddPCR assay compared with the synchronous blood culture. Secondary outcomes include the mortality rates and the mean Sequential Organ Failure Assessment (SOFA) score at follow-up time points, the length of stay in the hospital, the time to directed antimicrobial therapy, duration of broad-spectrum antibiotic use, and the EQ-5D-5L score on day 90. DISCUSSION: It is the first multicenter pragmatic RCT to explore the diagnostic efficacy and clinical impact of the ddPCR assay in patients with suspected sepsis, taking advantage of both RCT's ability to establish causality and the feasibility of pragmatic approaches in real-world studies (RWS). This trial will help us to get a comprehensive view of the assay's capacity for precise diagnosis and treatment of sepsis. It has the potential to monitor the pathogen load change and to guide the antimicrobial therapy, making a beneficial impact on the prognosis of sepsis patients. TRIAL REGISTRATION: ClinicalTrial.gov, NCT05190861. Registered January 13, 2022-'Retrospectively registered', https://clinicaltrials.gov/ct2/show/NCT05190861 .

A predictive model of macrosomic birth based upon real-world clinical data from pregnant women.

BACKGROUND: Fetal macrosomia is associated with an increased risk of several maternal and newborn complications. Antenatal predication of fetal macrosomia remains challenging. We aimed to develop a nomogram model for the prediction of macrosomia using real-world clinical data to improve the sensitivity and specificity of macrosomia prediction. METHODS: In the present study, we performed a retrospective, observational study based on 13,403 medical records of pregnant women who delivered singleton infants at a tertiary hospital in Shanghai from 1 January 2018 through 31 December 2019. We split the original dataset into a training set (n = 9382) and a validation set (n = 4021) at a 7:3 ratio to generate and validate our model. The candidate variables, including maternal characteristics, laboratory tests, and sonographic parameters were compared between the two groups. A univariate and multivariate logistic regression was carried out to explore the independent risk factors for macrosomia in pregnant women. Thus, the regression model was adopted to establish a nomogram to predict the risk of macrosomia. Nomogram performance was determined by discrimination and calibration metrics. All the statistical analysis was analyzed using R software. RESULTS: We compared the differences between the macrosomic and non-macrosomic groups within the training set and found 16 independent risk factors for macrosomia (P < 0.05), including biparietal diameter (BPD), head circumference (HC), femur length (FL), amniotic fluid index (AFI) at the last prenatal examination, pre-pregnancy body mass index (BMI), and triglycerides (TG). Values for the areas under the curve (AUC) for the nomogram model were 0.917 (95% CI, 0.908-0.927) and 0.910 (95% CI, 0.894-0.927) in the training set and validation set, respectively. The internal and external validation of the nomogram demonstrated favorable calibration as well as discriminatory capability of the model. CONCLUSIONS: Our model has precise discrimination and calibration capabilities, which can help clinical healthcare staff accurately predict macrosomia in pregnant women.

Frailty index is useful for predicting postoperative morbidity in older patients undergoing gastrointestinal surgery: a prospective cohort study.

BACKGROUND: Many assessment tools have been used to identify frail surgical patients. This study was designed to explore the prediction value of the frailty index (FI) for postoperative morbidity in older patients undergoing elective gastrointestinal surgery. METHODS: Between January 2019 and September 2020, we conducted a prospective study in our hospital, and patients aged over 65 years were enrolled. The FI assessment was conducted by two specialist nurses based on the 38-item scale, and patients were considered frail if the FI score was ≥ 0.25. The primary outcome was 30-day postoperative morbidity. Univariable and multivariable analyses were used to find the risk factors related to postoperative morbidity. RESULTS: A total of 246 consecutive patients were enrolled, for whom the median age was 72.0 [interquartile range (IQR): 67.0-77.0] years old, and 175 (71.1%) were male. Of these, 47 (19.1%) were frail. Patients with frailty were associated with older age (p < 0.001), higher American Society of Anesthesiologists (ASA) grade (p = 0.006), lower body mass index (p = 0.001), lower albumin (p = 0.003) and haemoglobin (p < 0.001) levels, increased blood loss (p = 0.034), increased risk of postoperative morbidity (p < 0.001), increased median length of stay (p = 0.017), and increased median postoperative hospital stay (p = 0.003). Multivariable analysis revealed that ASA grade [odds ratio (OR): 2.59, 95% confidence interval (CI) 1.19-5.64, p = 0.016], FI score (OR 7.68, 95% CI 3.19-18.48, p < 0.001) and surgical complexity (OR 22.83, 95% CI 5.46-95.51, p < 0.001) were independent predictors of 30-day postoperative morbidity. However, for patients with major surgery, FI score was the only independent predictor (OR 8.67, 95% CI 3.23-23.25, p < 0.001). CONCLUSION: Frailty was associated with adverse perioperative outcomes, and the 38-item FI scale was a useful frailty screening tool for older patients undergoing elective gastrointestinal surgery. For patients with major surgery, frailty was a more reliable predictor of postoperative 30-day morbidity than age and ASA grade.

Relaxin inhibits renal fibrosis and the epithelial-to-mesenchymal transition via the Wnt/ß-catenin signaling pathway.

Renal fibrosis is a common characteristic and the final pathological mechanism of chronic kidney disease (CKD). Although CKD remains incurable, inhibition of renal fibrosis is beneficial to inhibit the CKD process. Relaxin alleviates renal fibrosis in some experimental models, but its mechanism remains unclear. In the following, we studied the regulatory effect of relaxin on epithelial-mesenchymal transition (EMT) after unilateral ureteral obstruction (UUO). Our results demonstrate that relaxin could downregulate Wnt/ß-catenin signaling and decrease EMT, thus protecting against loss of transporters in tubular epithelial cells (TECs) and abrogate renal interstitial fibrosis following UUO. We confirmed that relaxin can downregulate Wnt/ß-catenin signaling and decrease EMT in NRK52E, thus abrogating G2 cell cycle arrest in vitro experiments. Therefore, a novel mechanism by which relaxin is antifibrotic is that relaxin regulates the EMT program of TECs via Wnt/ß-catenin signaling pathway. The inhibition of EMT contributes to protecting the functional capabilities of TECs and promoting the regeneration of TECs.

Comparison of alveolar bone width and sagittal tooth angulation of maxillary central incisors in Class I and Class III canine relationships: a retrospective study using CBCT.

BACKGROUND: Canine relationship is a key reference identifying anterior malocclusion and an important implication for evaluating preimplantation bone morphology at maxillary esthetic zone. This study aimed to compare the differences of maxillary central incisor-related measurements (alveolar bone thickness and tooth sagittal angulation) between Class I and Class III canine relationship and further explore the risk factors for immediate implant placement in the anterior maxilla based on cone beam computed tomography (CBCT) data. METHODS: CBCT digital imaging and communications in medicine (DICOM) files of 107 patients (54 with Class I canine relationship and 53 with Class III canine relationship) were collected and the alveolar bone thickness at mid-root (mid-root buccal thickness/MBT; palatal/MPT), apical regions (apical buccal thickness/ABT; palatal/APT) and sagittal angulation (SA) of the maxillary central incisor at the examined side were measured on the mid-sagittal observation plane. Descriptive statistical analysis and frequency distributions of the measurements based on Class I or Class III canine relationship were established. Statistical analyses were performed using Fisher's exact test, independent samples t test and Pearson correlation test with the significance level set at p < 0.05. RESULTS: The frequency distributions of maxillary central incisors' MPT, ABT, APT and SA showed significant differences between Class I and Class III canine relationships (p = 0.030, 0.024, 0.000 and 0.000, respectively). MPT (2.48 ± 0.88 mm vs. 3.01 ± 1.04 mm, p = 0.005), APT (6.79 ± 1.65 mm vs. 8.47 ± 1.93 mm, p = 0.000) and SA (12.23 ± 5.62° vs. 16.42 ± 4.49°, p = 0.000) were significantly smaller in patients with Class III canine relationship. Moreover, SA showed a strong positive correlation with APT (R = 0.723, p = 0.000) and a moderate negative correlation with ABT (R = - 0.554, p = 0.000). CONCLUSIONS: In populations with Class III canine relationship, maxillary central incisors were significantly more labially inclined and have a thinner palatal bone plate at the apex compared with Class I relationship. Clinicians should avoid palatal perforation during immediate implantation at sites of originally protrusive maxillary incisors.

Negative Pressure Wound Therapy With Chymotrypsin Irrigation: A Maximal Implant Retention Procedure Treating the Exposure/Infection of Titanium Mesh in Cranioplasty.

This preliminary study aims to investigate the effects of a maximal implant retention procedure. The authors retrospectively reviewed the use of negative pressure wound therapy with chymotrypsin irrigation treating implant infection/exposure in titanium mesh cranioplasty by comparing patients with titanium mesh totally retained, partially removed, or totally removed according to the evaluation during the surgery. Negative pressure wound therapy with chymotrypsin irrigation was applied 5 days after the surgery. The negative pressure was set at -125 to -150 mmHg. A total of 21 patients were included, 4 patients treated with titanium mesh totally removed; 3 patients treated with titanium mesh partially removed; and 14 patients treated with U-shape debridement with titanium mesh preserved completely. However, 1 patient in the U-shape group required a second debridement to remove all implant. Negative pressure wound therapy with chymotrypsin irrigation is a novel procedure and could be used to treat implant-related infection without the exchange of implant.

[Effect of Xiaoyao San on OVX combined with CUS anxiety and depression model rats based on hippocampal microglia M1 polarization].

To investigate the anti-anxiety and anti-depression effect and mechanism of Xiaoyao San on rats with ovariectomy(OVX) combined with chronic unpredictable stress(CUS) model. The model of perimenopausal depression was established by OVX and CUS; the level of anxiety and depression was evaluated by open field test; the levels of interleukin-1ß(IL-1ß) and interleukin-6(IL-6) mRNA in rat hippocampus were detected by Real-time qPCR; double staining immunofluorescence was used to detect the expression of ionized calcium binding adaptor molecule-1(Iba-1) and inducible nitric oxide synthase(iNOS) in microglia of rat dentate gyrus(DG); Western blot was used to detect the protein expression of Iba-1 and iNOS of microglia in DG region of rat hippocampus. The results showed that in the model group, the number of horizontal movement, the number of vertical movement and central residence time were significantly reduced, and the grooming time was significantly prolonged(P<0.05 or P<0.01); the levels of IL-1ß and IL-6 in hippocampus increased significantly(P<0.05); the number of positive cells with co-expression of Iba-1/iNOS of microglia cells in DG region of hippocampus increased; the expression levels of Iba-1 and iNOS protein in hippocampus were significantly increased(P<0.01), suggesting that microglia in DG region of hippocampus was activated and polarized toward M1 type in rats with stress. The high dose group of Xiaoyao San significantly increased the number of horizontal movement, vertical movement and central residence time of model rats(P<0.05 or P<0.01), and significantly down-regulated the levels of inflammatory factors IL-1ß and IL-6(P<0.05). Meanwhile, it reversed the activation and quantity change of microglia in hippocampus. Although the Xiaoyao San low dose group had no significant effect on the behavioral indicators in the open field test and the levels of IL-1ß and IL-6, they all showed a trend of improvement. Low dose Xiaoyao San significantly decreased iNOS protein level(P<0.05), and high dose Xiaoyao San significantly down-regulated the protein expression of Iba-1 and iNOS in hippocampus microglia(P<0.05 or P<0.01). In conclusion, Xiaoyao San can improve anxiety and depression-like behavior in OVX combined with CUS model rats, and its mechanism is related to its anti-inflammatory effect by inhibiting M1 polarization of hippocampal microglia.

Inhibition of curcumin on human lung adenocarcinoma LTEP-A2 cells and its mechanism.

This study was designed to explore the effects of curcumin on proliferation, migration and invasion of human lung adenocarcinoma LTEP-A2 cells and determine its mechanism. Human lung adenocarcinoma LTEP-A2 cell was cultured in vitro. After incubation with different concentrations of curcumin (5, 10, 15 µmol/L), the effects of curcumin on proliferation, migration and invasion of human lung adenocarcinoma LTEP-A2 cells were observed by MTT assay, wound healing and transwell assay. The expression levels of COX-2 and MMP-9 were detected by western blot. Compared with the blank control group, curcumin decreased the survival rate of LTEP-A2 cells, shorten the cell migration distance and decrease the number of LTEP-A2 cells penetrating membrane. The expression levels of COX-2 and MMP-9 were both down-regulated by curcumin. Curcumin can inhibit the proliferation, migration and invasion of human lung adenocarcinoma LTEP-A2 cells. The mechanism may be related to the down-regulation of COX-2 and MMP-9 expression.

TPD52L2 impacts proliferation, invasiveness and apoptosis of glioblastoma cells via modulation of wnt/ß-catenin/snail signaling.

Intratumoral heterogeneity greatly hinders efficiency of target therapy in glioblastoma (GBM). To decipher the underlying mechanisms of heterogeneity, patient-derived adult GBM cells were separately isolated from margins of T1 gadolinium enhancing tumor lesions (PNCs) and T1 gadolinium enhancing core lesions (ECs). Single clone culture was conducted in ECs and U87MG cell line to screen clones with distinct biological phenotypes. Single cell clones with diverse phenotypes were simultaneously separated from ECs and U87 cell line. PNCs, GCs(H) and U87(H) exhibited longer cellular protrusion than ECs, GCs(L) and U87(L), respectively. Cell strains with longer protrusion exhibited higher invasive ability and lower sensitivity to temozolomide (TMZ) and radiation. Subsequently, TPD52L2 was verified as the functional protein to regulate the cellular heterogeneity by the proteomics analysis. Downregulation of TPD52L2 enhanced cell invasion whereas inhibited cell proliferation rate and sensitivity to chemotherapy in vivo and in vitro, this condition was reversed when TPD52L2 was overexpressed. The invasiveness was facilitated by up-regulating CTNNB1/ß-catenin and SNAI1/Snail mediated EMT process. In addition, the clinical data of 88 GBM cases in our neurosurgery center was analyzed to reveal the influence of TPD52L2 in the prognosis of GBM. Low expression of TPD52L2 exacerbated prognosis of GBM patients received standard radiotherapy plus concomitant and adjuvant TMZ (Stupp strategy). Taken together, TPD52L2 is an important biomarker influencing GBM prognosis.

MLN64 deletion suppresses RANKL-induced osteoclastic differentiation and attenuates diabetic osteoporosis in streptozotocin (STZ)-induced mice.

Diabetes mellitus is known as a main cause to induce osteoporosis. However, the pathogenesis of osteoporosis induced by diabetes has not been fully understood. MLN64 is highly homologous to the steroidogenic acute regulatory (STAR) protein, sharing the highly conserved START domain and exhibiting various biological activities. In the study, we attempted to explore the role of MLN64 in osteoporosis progression through the in vitro and in vivo studies. At first, the in vitro study suggested that MLN64 was over-expressed during the osteoclast differentiation induced by receptor activator of NF-kB ligand (RANKL). MLN64-knockdown markedly reduced the number of TRAP-positive multinucleated cells induced by RANKL, along with the down-regulation of specific genes related to osteoclastogenesis, including tartrate-resistant acid phosphatase (TRAP), nuclear factor of activated T cells (NFATc1), cathepsin K (CTSK), tartrate resistant (ACP5) and c-FOS and up-regulation of transcriptional receptor runt related transcription factor 2 (Runx2) and osteopontin (OPN). In contrast, over-expressing MLN64 significantly promoted the production of TRAP-positive multinucleated cells triggered by RANKL. Moreover, RANKL exposure led to remarkable increase in inflammatory factors, including tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), IL-6, and MMP-1, which was attenuated by the knockdown of MLN64. Additionally, the expression of phosphorylated (p)IKKα, p-IκBα and p-nuclear factor (NF)-κB stimulated by RANKL was clearly alleviated by MLN64 silence. However, RANKL-induced inflammation was accelerated by MLN64 over-expression. Further, the in vivo study suggested that streptozotocin (STZ)-caused reduction in body weight of mice was improved by the loss of MLN64. STZ-induced diabetic mice exhibited higher levels of blood alkaline phosphatase (ALP) and TRAP, while being down-regulated in MLN64-knockout mice. MLN64 deletion reversed STZ-induced trabecular deleterious effects and stimulated bone remodeling. What's more, MLN64 knockout inhibited the expression levels of TRAP, RANKL and RANK in femoral heads of STZ mice, accompanied with the repression of inflammatory response. In conclusion, the results in our study suggested that MLN64 played a critical role in the meditation of osteoclastic differentiation, and its suppression alleviated diabetic osteoporosis in STZ-induced mice. Thus, MLN64 could be served as an essential target for developing effective therapeutic strategy to prevent diabetic osteoporosis.

Rhododendron oldhamii leaf extract improves fatty liver syndrome by increasing lipid oxidation and decreasing the lipogenesis pathway in mice.

Some members of Rhododendron genus are traditionally used as medicinal plants for arthritis, acute and chronic bronchitis, asthma, pain, inflammation, rheumatism, hypertension and metabolic diseases. To the best of our knowledge, there is no report on the protective effects of R. oldhamii leaf extract on non-alcoholic fatty liver disease (NAFLD) in vivo and in vitro. In this study, the effects of R. oldhamii leaf extract on inhibiting the free fatty acid (FFA)-induced accumulation of fat in HepG2 cells and on improving fatty liver syndrome in mice with high fat diet (HFD)-induced NAFLD were investigated. For the in vitro assay, HepG2 cells were treated with FFAs (oleate/palmitate = 2:1) with or without treatment with R. oldhamii leaf ethyl acetate (EtOAc) fraction to observe lipid accumulation using Nile red and oil red O stains. For the in vivo assay, C57BL/6 mice were randomly assigned to three groups (n = 5), including the normal diet group, the HFD group and the HFD+EtOAc group. After 11 weeks, body weight, serum biochemical indices and the mRNA expressions of the liver tissue, as well as the outward appearance, weight and histopathological analysis of liver and adipose tissues were evaluated. Among the fractions derived from R. oldhamii leaf, the EtOAc fraction exhibited a strong fat-accumulation inhibitory activity. Following reverse-phase high-performance liquid chromatography (HPLC), four specific phytochemicals, including (2R, 3R)-astilbin (AS), hyposide (HY), guaijaverin (GU) and quercitrin (QU), were isolated and identified from the EtOAc fraction of R. oldhamii leaf extract. Among them, AS and HY showed excellent fat-accumulation inhibitory activity. Thus, the EtOAc fraction of R. oldhamii leaf and its derived phytochemicals have great potential in preventing FFA-induced fat accumulation. In addition, the EtOAc fraction of R. oldhamii leaf significantly improved fatty liver syndrome and reduced total cholesterol (TC) and triglyceride (TG) in HFD-induced NAFLD mice at a dosage of 200 mg/kg BW. These results demonstrated that the methanolic extracts from R. oldhamii leaf have excellent inhibitory activities against fat accumulation and anti-NAFLD activities and thus have great potential as a natural health product.