Identification, validation and biological characterisation of novel glioblastoma tumour microenvironment subtypes: implications for precision immunotherapy.

BACKGROUND: New precision medicine therapies are urgently required for glioblastoma (GBM). However, to date, efforts to subtype patients based on molecular profiles have failed to direct treatment strategies. We hypothesised that interrogation of the GBM tumour microenvironment (TME) and identification of novel TME-specific subtypes could inform new precision immunotherapy treatment strategies. MATERIALS AND METHODS: A refined and validated microenvironment cell population (MCP) counter method was applied to >800 GBM patient tumours (GBM-MCP-counter). Specifically, partition around medoids (PAM) clustering of GBM-MCP-counter scores in the GLIOTRAIN discovery cohort identified three novel patient clusters, uniquely characterised by TME composition, functional orientation markers and immune checkpoint proteins. Validation was carried out in three independent GBM-RNA-seq datasets. Neoantigen, mutational and gene ontology analysis identified mutations and uniquely altered pathways across subtypes. The longitudinal Glioma Longitudinal AnalySiS (GLASS) cohort and three immunotherapy clinical trial cohorts [treatment with neoadjuvant/adjuvant anti-programmed cell death protein 1 (PD-1) or PSVRIPO] were further interrogated to assess subtype alterations between primary and recurrent tumours and to assess the utility of TME classifiers as immunotherapy biomarkers. RESULTS: TMEHigh tumours (30%) displayed elevated lymphocyte, myeloid cell immune checkpoint, programmed cell death protein 1 (PD-1) and cytotoxic T-lymphocyte-associated protein 4 transcripts. TMEHigh/mesenchymal+ patients featured tertiary lymphoid structures. TMEMed (46%) tumours were enriched for endothelial cell gene expression profiles and displayed heterogeneous immune populations. TMELow (24%) tumours were manifest as an 'immune-desert' group. TME subtype transitions upon recurrence were identified in the longitudinal GLASS cohort. Assessment of GBM immunotherapy trial datasets revealed that TMEHigh patients receiving neoadjuvant anti-PD-1 had significantly increased overall survival (P = 0.04). Moreover, TMEHigh patients treated with adjuvant anti-PD-1 or oncolytic virus (PVSRIPO) showed a trend towards improved survival. CONCLUSIONS: We have established a novel TME-based classification system for application in intracranial malignancies. TME subtypes represent canonical 'termini a quo' (starting points) to support an improved precision immunotherapy treatment approach.

Effect of transauricular nerve stimulation on perioperative pain: a single-blind, analyser-masked, randomised controlled trial.

BACKGROUND: Activation of central autonomic pathways, including those regulating the arterial baroreflex, might reduce acute pain. We tested the hypothesis that transcutaneous auricular nerve stimulation (TAN) reduces pain after orthopaedic trauma surgery through autonomic modulation. METHODS: A total of 86 participants aged >18 yr were randomly assigned to 50 min of either sham or active bilateral TAN, undertaken before, and again 24 h after, surgery for orthopaedic trauma. The primary outcome was absolute change in pain 24 h postoperatively, comparing the 100 mm visual analogue scale (VAS) before and after TAN. Secondary outcomes included the minimal clinically important difference in pain (>10 mm increase or reduction in VAS) before/after surgery, using intention-to-treat analysis. Holter monitoring, the analysis of which was masked to allocation, quantified autonomic modulation of heart rate. RESULTS: From June 22, 2021 to July 7, 2022, 79/86 participants (49 yr; 45% female) completed TAN before and after surgery. For the primary outcome, the mean reduction in VAS was 19 mm (95% confidence interval [CI]: 12-26) after active TAN (n=40), vs 10 mm (95% CI: 3-17) after sham TAN (n=39; P=0.023). A minimally clinically important reduction in postoperative pain occurred in 31/40 (78%) participants after active TAN, compared with 15/39 (38%) allocated to sham TAN (odds ratio 5.51 [95% CI: 2.06-14.73]; P=0.001). Only active TAN increased heart rate variability (log low-frequency power increased by 0.19 ms2 [0.01-0.37 ms2]). Prespecified adverse events (auricular skin irritation) occurred in six participants receiving active TAN, compared with two receiving sham TAN. CONCLUSION: Bilateral TAN reduces perioperative pain through autonomic modulation. These proof-of-concept data support a non-pharmacological, generalisable approach to improve perioperative analgesia.

[Applications and challenges of large language models in critical care medicine].

The rapid development of big data methods and technologies has provided more and more new ideas and methods for clinical diagnosis and treatment. The emergence of large language models (LLM) has made it possible for human-computer interactive dialogues and applications in complex medical scenarios. Critical care medicine is a process of continuous dynamic targeted treatment. The huge data generated in this process needs to be integrated and optimized through models for clinical application, interaction in teaching simulation, and assistance in scientific research. Using the LLM represented by generative pre-trained transformer ChatGPT can initially realize the application in the diagnosis of severe diseases, the prediction of death risk and the management of medical records. At the same time, the time and space limitations, illusions and ethical and moral issues of ChatGPT emerged as the times require. In the future, it is undeniable that it may play a huge role in the diagnosis and treatment of critical care medicine, but the current application should be combined with more clinical knowledge reserves of critical care medicine to carefully judge its conclusions.

The value of first-line chemotherapy and targeted therapy in the treatment of breast cancer.

OBJECTIVE: To investigate the value (survival benefit and cost) of first-line chemotherapy and targeted therapy in breast cancer at a population level. METHODS: Based on guideline recommendations, a model of optimal utilisation was constructed for first-line chemotherapy and targeted therapy in breast cancer, calculating the survival benefit and average cost of all regimens recommended for each treatment indication at 5 years and at 10 years. RESULTS: Survival benefits from chemotherapy and targeted therapy differ markedly depending on the treatment indications. The cost per life-year gained at 5 years is $38,044 for stages I and II, $33,749 for stage III and $ 151,668 for patients presenting with stage IV breast cancer. The cost per life-year gained at 10 years is $ 13,587 for early breast cancer. The most expensive chemotherapy indication in breast cancer is the treatment of metastatic HER2-positive breast cancer costing $330,978 per LYG for a survival benefit of 11% at 5 years falling to zero survival benefit at 10 years. CONCLUSION: There are large differences in value between the different indications for first-course chemotherapy and targeted therapy in the treatment of breast cancer that should be considered when pricing cancer drugs.

Robust, tunable, and high purity triggered single photon source at room temperature using a nitrogen-vacancy defect in diamond in an open microcavity.

We report progress in the development of tunable room temperature triggered single photon sources based on single nitrogen-vacancy (NV) centres in nanodiamond coupled to open access optical micro-cavities. The feeding of fluorescence from an NV centre into the cavity mode increases the spectral density of the emission and results in an output stream of triggered single photons with spectral line width of order 1 nm, tunable in the range 640 - 700 nm. We record single photon purities exceeding 96% and estimated device efficiencies up to 3%. We compare performance using plano-concave microcavities with radii of curvature from 25 µm to 4 µm and show that up to 17% of the total emission is fed into the TEM00 mode. Pulsed Hanbury-Brown Twiss (HBT) interferometry shows that an improvement in single photon purity is facilitated due to the increased spectral density.

[Pigmented extramammary Paget's disease accompanied with condyloma acuminatum: a case report].

Pigmented extramammary Paget's disease (PEMPD) is an uncommon intraepithelial adenocarcinoma and a rare variant of Paget's disease, characterized as a superficial pigmented scaly macule clinically and an increased number of melanocytes scattered between the Paget's cells histologically. So it may be confused clinically and histologically with melanocytic tumors, dermatitis and other dermatoses. Different therapeutic attitudes are required in this case of adenocarcinoma in situ as opposed to melanoma and dermatitis. Condyloma acuminatum (CA) is a common sexually transmitted disease caused by human papilloma virus infection, which is also called as genital warts. In this article, we first reported a case of a 65-year-old Chinese man who had pigmented extramammary Paget's disease complicated with CA. This patient presented with verrucous papules on the scrotum for 3.5 years, infiltrative erythema with itch on the mons pubis for 3 years, and scrotum and penis involved gradually for 4 months. Physical examination showed a 8 cm×10 cm dark red patch on the upper part of the scrotum, penis and mons pubis, as well as few maculopapules and nodules. Histopathologic examination of the lesion on the scrotum revealed a focus of Paget's disease, characterized by the presence of large round cells with abundant pale or granular/dusty cytoplasm, pleomorphic vesicular nuclei and prominent nucleoli (Paget's cells), while the histology of the verrucous lesion was consistent with CA. Immunohistochemistry was performed, which showed diffuse positive staining with CK, CEA, PAS, CK20, EMA, CK7, and Ki-67 (40%), HER2 in Paget's cells and negative with P53, P16, CK5/6, S100, MelanA, HMB45, estrogen receptor, progesterone receptor, and gross cystic disease flid protein 15 (GCDFP15). Human papillomavirus-11 (HPV-11) was positive by genotyping using gene amplification in the lesion of scrotum. According to clinical features and laboratory findings, a diagnosis of PEMPD complicated with CA was made. Local excision of the lesion was performed and sent for histological examination, with all margins clear of tumor. Both aforementioned diseases often occur in the vulva. Even so, it has been rarely reported coexisting of the above two diseases, of which the clinical significance and association are also unclear. In this article, we also reviewed the literature relating to PEMPD, and on this basis, the profile of this disease is discussed including its pathogenesis, clinical manifestation, diagnosis, treatment and advances. Due to PEMPD occasionally accompanied with an underlying carcinoma, it's essential to make an accurate diagnosis. Besides, review of the literature reveals that pigmented variant of Paget's disease could be initially misdiagnosed as melanocytic tumors and other dermatoses unless the entity is considered in the differential diagnosis and additional confirmatory studies are performed.

[Acute respiratory failure due to Pneumocystis pneumonia in connective tissue disease patients: clinical manifestation and prognostic factors related to hospital mortality].

Objective: To investigate the clinical manifestations and prognostic factors of hospital death in connective tissue disease patients with acute respiratory failure caused by pneumocystis pneumonia (PCP) admitted to two medical intensive care units(MICU). Methods: A retrospective review was conducted for all connective tissue disease (CTD) patients with acute respiratory failure from PCP in MICU of 2 academic medical centers between 2010 and 2015. The patients were divided into survivors and non-survivors. Demographic and clinical data, including laboratory, radiological and microbiological findings, as well as therapy, clinical course, mortality and prognostic factors of hospital mortality were included in the analysis. Logistic regression models were used to determine the effect of prognostic factors on hospital death after adjusting for covariates of which the p values were less than 0.1. Results: A total of 41 patients with connective tissue disease were identified. The PaO(2)/FiO(2) ratio (PFR) on ICU admission was 120 mmHg(55-180 mmHg, 1 mmHg=0.133 kPa). Common clinical features included dyspnea (90.2%, 37/41), fever (87.8%, 36/41) and dry cough(65.9%, 30/41). 58.5%(24/41) and 17.1%(7/41) patients were co-infected by CMV and aspergillus, respectively. The overall mortality rate was 75.6%(31/41). Compared with survivors, the age, APACHEⅡ score and incidence of barotrauma in non-survivors were higher (39±17 vs 58±15, t=3.018, P=0.002), (15±6 vs 19±5, t=2.528, P=0.019), (0/10 vs 12/31, χ(2)=5.473, P=0.021), while PFR on ICU admission was lower in non-survivors (172±68 vs 116±49, t=-1.893, P=0.007). Logistic analysis showed that PFR on ICU admission was the independent risk factor for hospital death (OR=1.004, 95%CI: 1.002-1.006, P=0.048). Conclusions: Mortality rate among patients with acute respiratory failure secondary to CTD related PCP is still high, and the poor prognostic factors of hospital mortality included PFR on ICU admission and barotrauma.

Epigallocatechin gallate inhibits the growth of salivary adenoid cystic carcinoma cells via the EGFR/Erk signal transduction pathway and the mitochondria apoptosis pathway.

ACC is one of the most malignant tumors in salivary gland, and of poor prognosis. A critical role in ACC development and progression is played by EGFR family members including EGFR. EGCG, a low molecular weight polyphenol contained in green tea, has broad anticancer properties, but whether EGCG regulates activity of ACC is unknown. In the present study, the effects of EGCG were investigated in vitro with particular attention to the pathway of EGFR/Erk and mitochondria apoptosis in SACC-83 cell lines. The results of MTS assay and flow cytometry demonstrated that EGCG (20-80 µM) could inhibit proliferation and promote apoptosis of SACC-83 cells. Furthermore, by Western blotting with antibodies specific for EGFR, Erk 1/2 (p-Erk 1/2), Mek (p-Mek), Bcl-2, and Bax, it was demonstrated that EGCG could reduce the expression of EGFR, inhibit phosphorylation of Erk 1/2 and Mek, downregulate Bcl-2, and upregulate Bax. In addition, it was also shown that EGCG could inhibit mRNA expression of P90 RSK by RT-PCR. In conclusion, the results suggest that EGCG might be a potential therapeutic or adjuvant strategy for the treatment of patients with ACC, by inhibiting proliferation and inducing the apoptosis of the tumor cells.

[The 455th case: swollen leg, jaundice and mental disturbance].

A 17-year-old young man with a history of swollen leg and intermittent jaundice was presented to Peking Union Medical College Hospital with acute fever and mental disturbance. He developed deep venous thrombosis, acute myocardial infarction and plantar skin necrosis during the past four years, and was presented with an acute episode of fever, thrombocytopenia, acute kidney injury, acute myocardial infarction, mental disturbance, and obstructive jaundice. Laboratory tests showed schistocytes on peripheral blood smear.High titer of antiphospholipid antibodies was detected.Strikingly, the activity of a disintegrin and metalloprotease with a thrombospondin type 1 motif, member 13 (ADAMTS13)was significantly decreased without the production of inhibitors. Images indicated stenosis of the common bile duct, common hepatic duct, and cystic duct, which caused dilation of bile ducts and the gall bladder. Corticosteroids and anticoagulation therapy were effective at first, but the disease relapsedonce the corticosteroids tapered down. Plasma exchange was administrated for 17 times, which was effective temporarily during this episode. Methylprednisolone pulse therapy, intravenous immunoglobulin, rituximab, anticoagulation therapy, and bile drainage, were all tried but still could not control the disease. The patient's family agreed to withdraw treatment after he developed septic shock.

[Left atrial appendage volume is a valuable predictor of atrial fibrillation recurrence after radiofrequency catheter ablation].

Objective: To investigate the association between the left atrial appendage (LAA) volume and atrial fibrillation (AF) recurrence after radiofrequency catheter ablation. Methods: We prospectively enrolled sixty-two patients with AF (40 cases with paroxysmal AF, 22 cases with persistent AF) who successfully underwent a first AF catheter ablation and had performed contrast-enhanced cardiac computed tomography (CT) prior to the procedure to measure LAA volumes in our hospital from January 2012 to August 2015. Circumferential pulmonary vein isolation was performed under the guidance of three-dimension mapping system (CARTO system). Linear ablation or ablation of complex fractioned atrial electrograms was also undertaken if necessary. All patients were followed up at the 3rd, 6th and 12th months after ablation by 24-hour ambulatory Holter monitoring, and were divided into the non-recurrence group (n=42) and the AF recurrence group (n=20). Univariate and multivariate Cox proportional hazards regression analysis were used to assess the factors related to AF recurrence. The receiver operating characteristic (ROC) curve was calculated to assess the best cut-off value of LAA volume to predict AF recurrence. Kaplan-Meier method was used to evaluate the rate of freedom from AF recurrence. Results: Mean LAA volume in all patients was (9.5±3.6)ml. AF recurrence occurred in 20 patients (32%) during the follow-up period. The LAA volume was significantly larger in the AF recurrence group than in the non-recurrence group ((11.5±3.8)ml vs. (8.3±3.1)ml, P=0.002). In the univariate regression analysis, LAA volume (HR=1.36, 95%CI 1.14-1.82, P<0.001), persistent AF (HR=4.43, 95%CI 1.52-12.06, P<0.001) and hypertension (HR=1.61, 95%CI 1.13-2.04, P=0.041) were risk factors of AF recurrence. However, multivariate regression analysis revealed that LAA volume (HR=1.32, 95%CI 1.12-1.51, P<0.001) and persistent AF (HR=4.22, 95% CI 1.48-11.05, P<0.001) were independent predictors for AF recurrence after ablation. The receiver operating characteristic (ROC) curve analysis revealed that a LAA volume >8.80 ml was associated with AF recurrence after ablation (sensitivity: 94% and specificity: 66%, area under the curve=0.76). Kaplan-Meier analysis showed a lower rate free from AF recurrence in the group with LAA volume >8.80 ml (P<0.001). Conclusion: Larger LAA volume is associated with AF recurrence after catheter ablation in patients with AF. A LAA volume greater than 8.80 ml could be used to predict AF recurrence after ablation.

[Clinical features of acute respiratory failure due to pneumocystis pneumonia in non-HIV immunocompromised patients].

Objective: To examine the clinical features of patients with acute respiratory failure (ARF) caused by pneumocystis pneumonia (PCP) admitted into two medical intensive care units (ICU) in non- human immunodeficiency virus (HIV) infected immunocompromised patients. Methods: A retrospective review was conducted among 92 non-HIV patients with ARF caused by PCP in medical ICU of Peking Union Medical College Hospital and China-Japan Friendship Hospital between Jan 2010 and Dec 2015. Patient characteristics, clinical presentation, laboratory and radiological findings, complications, as well as therapy and mortality were included in the analysis. Results: All patients were immunocompromised before PCP, among which 69.6% (64/92) patients were suffered from autoimmune disease. The diagnosis of PCP was made by the identification of P. jiroveci organisms with Giemsa or polymerase chain reaction in specimens of bronchoalveolar lavage, sputum or tracheal aspiration. The Acute Physiology and Chronic Health Evaluation (APACHE) Ⅱ was high (19±5) and the partial pressure of oxygen/ fraction of inspiration oxygen(PaO2/FiO2) ratio was low[(139.6±65.4) mmHg]on ICU admission, with all patients diagnosed as acute respiratory failure during ICU stay. Radiologic findings showed bilateral diffused ground glass opacity (94.6%, 87/92). All patients received Compound Sulfamethoxazole (SMZ/TMP) and only 3.3% (3/92) patients were not given conjunctive corticosteroid. 57.6% (53/92) and 21.7% (20/92) patients were coinfected by cytomegalovirus (CMV) and aspergillos. Invasive ventilatory support was required in 90% (81/90) patients. 22% (18/82) patients used non-invasive positive pressure ventilation (NPPV) on ICU admission but most of them (83.3%, 15/18) failed and switched to invasive positive pressure ventilation (IPPV). Median ICU and hospital length of stay were 11 and 17 days, respectively. The overall hospital mortality rate was 76.1% (70/92). Conclusions: Among patients with ARF secondary to non-HIV related PCP, autoimmune system diseases are the most common primary diagnosis. The clinical manifestations were severe and coinfections are common, with poor prognosis.

Rearrangement of a large novel Pseudomonas aeruginosa gene island in strains isolated from a patient developing ventilator-associated pneumonia.

Bacterial gene islands add to the genetic repertoire of opportunistic pathogens. Here, we perform comparative analyses of three Pseudomonas aeruginosa strains isolated sequentially over a 3-week period from a patient with ventilator-associated pneumonia (VAP) who received clindamycin and piperacillin-tazobactam as part of their treatment regime. While all three strains appeared to be clonal by standard pulsed-field gel electrophoresis, whole-genome sequencing revealed subtle alterations in the chromosomal organization of the last two strains; specifically, an inversion event within a novel 124-kb gene island (PAGI 12) composed of 137 open reading frames [ORFs]. Predicted ORFs in the island included metabolism and virulence genes. Overexpression of a gene island-borne putative ß-lactamase gene was observed following piperacillin-tazobactam exposure and only in those strains that had undergone the inversion event, indicating altered gene regulation following genomic remodeling. Examination of a separate cohort of 76 patients with VAP for integration at this tRNA(lys) recombination site demonstrated that patients exhibiting evidence of integration at this site had significantly higher 28-day mortality. These findings provide evidence that P. aeruginosa can integrate, rapidly remodel, and express exogenous genes, which likely contributes to its fitness in a clinical setting.

Genome-wide discovery of genetic variants affecting tamoxifen sensitivity and their clinical and functional validation.

BACKGROUND: Beyond estrogen receptor (ER), there are no validated predictors for tamoxifen (TAM) efficacy and toxicity. We utilized a genome-wide cell-based model to comprehensively evaluate genetic variants for their contribution to cellular sensitivity to TAM. DESIGN: Our discovery model incorporates multidimensional datasets, including genome-wide genotype, gene expression, and endoxifen-induced cellular growth inhibition in the International HapMap lymphoblastoid cell lines (LCLs). Genome-wide findings were further evaluated in NCI60 cancer cell lines. Gene knock-down experiments were performed in four breast cancer cell lines. Genetic variants identified in the cell-based model were examined in 245 Caucasian breast cancer patients who underwent TAM treatment. RESULTS: We identified seven novel single-nucleotide polymorphisms (SNPs) associated with endoxifen sensitivity through the expression of 10 genes using the genome-wide integrative analysis. All 10 genes identified in LCLs were associated with TAM sensitivity in NCI60 cancer cell lines, including USP7. USP7 knock-down resulted in increasing resistance to TAM in four breast cancer cell lines tested, which is consistent with the finding in LCLs and in the NCI60 cells. Furthermore, we identified SNPs that were associated with TAM-induced toxicities in breast cancer patients, after adjusting for other clinical factors. CONCLUSION: Our work demonstrates the utility of a cell-based model in genome-wide identification of pharmacogenomic markers.

[Evaluation of human milk feeding in hospitalized very low and extremely low birth weight infants].

Objective: To investigate the current situation of human milk (HM) feeding in hospitalized very low and extremely low birth weight infants. Methods: The study retrospectively extracted the data of 601 infants with birth weight <1 500 g, and admitted within 24 hours after birth to the Neonatal Intensive Care Unit of Nanjing Maternity and Child Health Care Hospital from January 2016 to December 2018. The infants were grouped into exclusive mother's-own-milk (MOM) group, donor human milk (DHM) group (partial or none MOM), and mixed (HM and formula) feeding group according to the feeding strategy. Qualitative and quantitative variables in the three groups were compared with One-way ANOVA, Kruskal-Wallis test, Chi-square test or Fisher exact test. Kappa and McNemar test were used for consistency testing. Results: Among the 601 infants (309 boys and 292 girls), 6 (1.0%) infants had never been fed with MOM. The gestational age and birth weight were (29.3±1.9) weeks and 1 260(1 115, 1 400) g in 601 infants. A total of 8 (1.3%) infants were grouped into MOM group, 542 (90.2%) were grouped into DHM group, and 51 (8.5%) were grouped into mixed feeding group. The percentage of enteral feedings with MOM in the stage of hospitalization 1-7 d, 8-14 d and 15-28 d were 73.6% (42.9%, 86.7%), 97.5% (78.6%, 100.0%) and 99.3% (93.0%, 100.0%), respectively (H=414.95, P<0.01), and the pairwise comparison suggested that the stage of hospitalization 1-7 d was the lowest (adjusted both P<0.05). The average weight adjusted daily dose of MOM were 9.7 (4.3, 18.2), 59.1 (26.5, 93.5) and 116.0 (60.3, 142.6) ml/(kg·d) in the stage of hospitalization 1-7 d, 8-14 d and 15-28 d, respectively (H=759.75, P<0.01), and the pairwise comparison suggested that the stage of hospitalization 1-7 d was the lowest (adjusted both P<0.05). The weight adjusted daily dose of MOM in exclusive MOM group, DHM and Mixed feeding group were 95.2 (40.0, 117.2), 82.9(53.6, 103.1) and 55.7 (16.6, 97.5) ml/(kg·d), respectively (H=10.78, P=0.005).Additionally, the percentage and weight adjusted daily dose of MOM showed a general consistency of 0.703 (P>0.05, Kappa=0.408). Conclusions: The rate of exclusive MOM feeding is low, especially during the first 7 days of hospitalization. The percentage of total enteral feedings with MOM and the average weight adjusted daily dose of MOM can well evaluate the situation of HM feeding during hospitalization quantitively.

Inflammation induction of Dickkopf-1 mediates chondrocyte apoptosis in osteoarthritic joint.

OBJECTIVE: Dysregulated Wnt signaling appears to modulate chondrocyte fate and joint disorders. Dickkopf-1 (DKK1) regulates the pathogenesis of skeletal tissue by inhibiting Wnt actions. This study examined whether DKK1 expression is linked to chondrocyte fate in osteoarthritis (OA). METHOD: Articular cartilage specimens harvested from nine patients with knee OA and from six controls with femoral neck fracture were assessed for DKK1, interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha), Bad, Bax, Bcl2 and caspase-3 expression by real time-polymerase chain reaction (RT-PCR) and immunohistochemistry. Apoptotic chondrocytes were detected by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end-labelling (TUNEL) and 4', 6-dianidino-2-phenylindole dihydrochloride (DAPI) staining. Human chondrocyte cultures were treated with recombinant IL-1beta and monoclonal DKK1 antibody to determine whether DKK1 impairs chondrocyte survival. RESULTS: Expression of DKK1 correlated with inflammatory cytokine levels (IL-1beta and TNF-alpha expressions), proapoptosis regulators (Bad and caspase-3 expressions) and TUNEL staining in OA cartilage tissues. The IL-1beta induced expressions of DKK1, Bax, Bad and caspase-3-dependent apoptosis of chondrocyte cultures. Neutralization of DKK1 by monoclonal DKK1 antibody significantly abrogated IL-1beta-mediated caspase-3 cleavage and apoptosis and reversed chondrocyte proliferation. Recombinant DKK1 treatment impaired chondrocyte growth and promoted apoptosis. By suppressing nuclear beta-catenin accumulation and Akt phosphorylation, DKK1 mediated IL-1beta promotion of chondrocyte apoptosis. CONCLUSION: Chondrocyte apoptosis correlates with joint OA. Expression of DKK1 contributes to cartilage deterioration and is a potent factor in OA pathogenesis. Attenuating DKK1 may reduce cartilage deterioration in OA.

Factors Associated With the Willingness of Clinical Health Care Professionals for Living Organ Donation.

Understanding the attitudes and willingness in regard to living organ donation of health care professionals is important for clinical practice and needs to be addressed. Thus, the objective of this study was to examine willingness and its influencing factors in regard to living organ donation in clinical health care professionals. METHODS: This study used a cross-sectional design. A total of 375 health care professionals from a regional teaching hospital in northern Taiwan were included in this study. RESULTS: Nearly 65% of the participants indicated a willingness to donate their living organ. Of these participants, 97.1% were willing to donate to family members, 80.8% were willing to donate to friends, and 78.9% were willing to donate to strangers. The predictors of willingness to engage in living organ donation were the desire to help others (odds ratio [OR] = 2.96; P < .01), positive attitude toward living organ donation (OR = 1.12; P < .01), financial support from the government (OR = 4.99; P < .01), and fewer physical concerns (OR = 0.97; P = .04). The willingness to donate a living organ was not associated with age, sex, religious belief, education level, participation in voluntary work, years of clinical work, type of profession, or knowledge about living organ donation. CONCLUSION: In general, health care professionals had a positive attitude toward and willingness to engage in living organ donation. It is hoped that the results of this study will serve as a referent framework for policymaking in regard to living organ donation and transplantation.

Acute myeloid leukemia transforms the bone marrow niche into a leukemia-permissive microenvironment through exosome secretion.

Little is known about how leukemia cells alter the bone marrow (BM) niche to facilitate their own growth and evade chemotherapy. Here, we provide evidence that acute myeloid leukemia (AML) blasts remodel the BM niche into a leukemia growth-permissive and normal hematopoiesis-suppressive microenvironment through exosome secretion. Either engrafted AML cells or AML-derived exosomes increased mesenchymal stromal progenitors and blocked osteolineage development and bone formation in vivo. Preconditioning with AML-derived exosomes 'primed' the animals for accelerated AML growth. Conversely, disruption of exosome secretion in AML cells through targeting Rab27a, an important regulator involved in exosome release, significantly delayed leukemia development. In BM stromal cells, AML-derived exosomes induced the expression of DKK1, a suppressor of normal hematopoiesis and osteogenesis, thereby contributing to osteoblast loss. Conversely, treatment with a DKK1 inhibitor delayed AML progression and prolonged survival in AML-engrafted mice. In addition, AML-derived exosomes induced a broad downregulation of hematopoietic stem cell-supporting factors (for example, CXCL12, KITL and IGF1) in BM stromal cells and reduced their ability to support normal hematopoiesis. Altogether, this study uncovers novel features of AML pathogenesis and unveils how AML cells create a self-strengthening leukemic niche that promotes leukemic cell proliferation and survival, while suppressing normal hematopoiesis through exosome secretion.