Short-term function and immune-protection of microencapsulated adult porcine islets with alginate incorporating CXCL12 in healthy and diabetic non-human primates without systemic immune suppression: A pilot study.

Replacement of insulin-producing pancreatic beta-cells by islet transplantation offers a functional cure for type-1 diabetes (T1D). We recently demonstrated that a clinical grade alginate micro-encapsulant incorporating the immune-repellent chemokine and pro-survival factor CXCL12 could protect and sustain the integrity and function of autologous islets in healthy non-human primates (NHPs) without systemic immune suppression. In this pilot study, we examined the impact of the CXCL12 micro encapsulant on the function and inflammatory and immune responses of xenogeneic islets transplanted into the omental tissue bilayer sac (OB; n = 4) and diabetic (n = 1) NHPs. Changes in the expression of cytokines after implantation were limited to 2-6-fold changes in blood, most of which did not persist over the first 4 weeks after implantation. Flow cytometry of PBMCs following transplantation showed minimal changes in IFNγ or TNFα expression on xenoantigen-specific CD4+  or CD8+  T cells compared to unstimulated cells, and these occurred mainly in the first 4 weeks. Microbeads were readily retrievable for assessment at day 90 and day 180 and at retrieval were without microscopic signs of degradation or foreign body responses (FBR). In vitro and immunohistochemistry studies of explanted microbeads indicated the presence of functional xenogeneic islets at day 30 post transplantation in all biopsied NHPs. These results from a small pilot study revealed that CXCL12-microencapsulated xenogeneic islets abrogate inflammatory and adaptive immune responses to the xenograft. This work paves the way toward future larger scale studies of the transplantation of alginate microbeads with CXCL12 and porcine or human stem cell-derived beta cells or allogeneic islets into diabetic NHPs without systemic immunosuppression.

Intrapleural transplantation of allogeneic pancreatic islets achieves glycemic control in a diabetic non-human primate.

Clinical islet transplantation has relied almost exclusively on intraportal administration of pancreatic islets, as it has been the only consistent approach to achieve robust graft function in human recipients. However, this approach suffers from significant loss of islet mass from a potent immediate blood-mediated inflammatory response (IBMIR) and a hypoxic environment. To avoid these negative aspects of the portal site, we explored an alternative approach in which allogeneic islets were transplanted into the intrapleural space of a non-human primate (NHP), treated with an immunosuppression regimen previously reported to secure routine survival and tolerance to allogeneic islets in NHP. Robust glycemic control and graft survival were achieved for the planned study period of >90 days. Our observations suggest the intrapleural space provides an attractive locale for islet transplantation due to its higher oxygen tension, ability to accommodate large transplant tissue volumes, and a lack of IBMIR-mediated islet damage. Our preliminary results reveal the promise of the intrapleural space as an alternative site for clinical islet transplantation in the treatment of type 1 diabetes.

In vivo induction of regulatory T cells by anti-CD45RB antibody causes transferable tolerance to discordant human xenogenic islets.

BACKGROUND: The treatment of diabetes by islet cell transplantation has become an accepted therapy, with transplantation of xenogeneic islet cells an attractive alternative to the problem. Previous studies in mice have demonstrated that anti-CD45RB induce immune tolerance in human pancreatic islet cells. The current study was to define the mechanism of action of anti-CD45RB induced nonspecific immune tolerance to heteroantigens. METHODS: A total of 1500 IEQ human islets were transplanted to diabetic B6µMT-/- mice, B6 mice, and µMT-/- diabetic mice undergoing thymectomy. These mice were treated short-term with doses of anti-CD45RB. CD4+Foxp3+Tregs were detected in the blood, peripheral lymphatic organs by flow cytometry, and immunohistochemistry. In addition, anti-CD25 mAb was administered to tolerant human islet cells B6µMT-/-mice. Mice then were transplanted with other human islet cells and received CD4+CD25+Tregs isolated from tolerant human islets mice to observe islet destruction. RESULTS: Anti-CD45RB treatment-induced tolerance to islets in both immunocompetent and B-cell-deficient mice (µMT-/- mice) by processes that were dependent on CD25+ Tregs, but not B cells. Anti-CD45RB treatment increased the number of CD4+Foxp3+Tregs cells. Anti-CD45RB treatment-induced immune tolerance that was antigen nonspecific, with Tregs playing an important role. Anti-CD45RB treatment-induced tolerance generated Tregs that could be transferred to another individual to manifest nonspecific immune tolerance. CONCLUSION: The results of the experiment suggest that anti-CD45RB induced tolerance to human islet xenografts is mediated by the proliferation of Tregs. These tolerogenic Tregs can be transferred to other mice and induce nonspecific immune tolerance.

Spectroscopy, Thermodynamics and Molecular Docking of Fraxinellone with DNA.

Fraxinellone is an important botanical lactone compound and has been demonstrated to have insecticidal activity. To provide theoretical support to the assessment on the safety of utilizing fraxinellone as a natural insecticidal agent, the interactions between fraxinellone and armyworm DNA, salmon sperm DNA and calf thymus DNA were investigated using UV-Vis absorption spectroscopy, isothermal titration calorimetry, and molecular docking. Results showed that there were two types of combinations between fraxinellone and three kinds of DNA. Type I combination had an equilibrium constant of combination (Ka1) of about 105 and binding sites (n1) of 0.40-0.70, while type II combination had an equilibrium constant of combination (Ka2) of 103 and binding sites (n2) of 1.35-3.15. Results of molecular docking showed that there were non-classical embedding type interactions between fraxinellone and three kinds of DNA, with the reaction taking place in small groove areas of the DNA structure, resulting in relatively weak interactive forces.

Band structure analysis of phononic crystals based on the Chebyshev interval method.

Due to environmental changes and manufacturing errors, uncertainties inherently exist in phononic crystals, especially in the material properties and geometric parameters. To handle the uncertainties with limited information, two different methods are proposed for band structure analysis of phononic crystals with uncertainties. One is the Monte Carlo method (MCM). The main shortcoming of it is the high computational cost. The other is the simplex Chebyshev polynomial expansion (SPCE) method. In addition, the computational efficiency of the SPCE method is much higher. In the SPCE method, the simplex Chebyshev is employed to estimate the band structures of phononic crystals. Meanwhile, the simplified incremental sampling strategy is introduced for the simplex Chebyshev to retain calculation accuracy and improve computational efficiency simultaneously. In the Chebyshev surrogate model, the samples yielded with the MCM are used to calculate the interval ranges of the band structures in phononic crystals. Three numerical examples, including a two-dimensional (2D) Helmholtz resonator phononic crystal, a 2D solid-solid phononic crystal, and a three-dimensional phononic crystal, are introduced to verify the effectiveness and efficiency of the proposed method.

Developmental changes in digestive enzyme activity in American shad, Alosa sapidissima, during early ontogeny.

In order to assess the digestive physiological capacity of the American shad Alosa sapidissima and to establish feeding protocols that match larval nutritional requirements, we investigated the ontogenesis of digestive enzymes (trypsin, amylase, lipase, pepsin, alkaline phosphatase, and leucine aminopeptidase) in larvae, from hatching to 45 days after hatching (DAH). We found that all of the target enzymes were present at hatching, except pepsin, which indicated an initial ability to digest nutrients and precocious digestive system development. Trypsin rapidly increased to a maximum at 14 DAH. Amylase sharply increased until 10 DAH and exhibited a second increase at 33 DAH, which coincided with the introduction of microdiet at 30 DAH, thereby suggesting that the increase was associated with the microdiet carbohydrate content. Lipase increased until 14 DAH, decreased until 27 DAH, and then increased until 45 DAH. Pepsin was first detected at 27 DAH and then sharply increased until 45 DAH, which suggested the formation of a functional stomach. Both alkaline phosphatase and leucine aminopeptidase markedly increased until 18 DAH, which indicated intestinal maturation. According to our results, we conclude that American shad larvae possess the functional digestive system before mouth opening, and the significant increases in lipase, amylase, pepsin, and intestinal enzyme activities between 27 and 33 DAH suggest that larvae can be successfully weaned onto microdiets around this age.

Hydrolysis mechanism of carbendazim hydrolase from the strain Microbacterium sp. djl-6F.

The carbendazim (MBC) hydrolyzing enzyme gene was cloned and heterologously expressed in Escherichia coli BL21 (DE3) from a newly isolated MBC-degrading bacterium strain Microbacterium sp. strain djl-6F. High performance liquid chromatography-mass spectrometry (HPLC-MS) analysis revealed that purified MheI-6F protein catalyzes direct hydrolysis of MBC into 2-aminobenzimidazole (2-AB) with a high turnover rate and moderate affinity (Km of 6.69µmol/L and kcat of 160.88/min) without the need for any cofactors. The optimal catalytic condition of MheI-6F was identified as 45°C, pH7.0. The enzymatic activity of MheI-6F was found to be diminished by metal ions, and strongly inhibited by sodium dodecyl sulfate (SDS). Through generating amino acid mutations in MheI-6F, Cys16 and Cys222 were identified as the catalytic groups that are essential for the hydrolysis of MBC. This is the first report on the biodegradation of MBC at the enzymatice level.

Stress and immune responses in skin of turbot (Scophthalmus maximus) under different stocking densities.

Fish skin and its mucus provide the first line of defense against chemical, physical and biological stressors, but little is known about the role of skin and its mucus in immune response to crowding stress. In the present study, we investigated the stress and immune responses in skin of turbot (Scophthalmus maximus) under different stocking densities. Turbot (average weight 185.4 g) were reared for 120 days under three densities: low density (LD), medium density (MD), and high density (HD). After 120 days, fish were weighed and sampled to obtain blood, mucus and skin tissues which were used for analyses of biochemical parameters and genes expression. The results showed HD treatment significantly suppressed growth and enhanced plasma cortisol and glucose levels (P < 0.05). In mucus, the activities of lysozyme (LZM), alkaline phosphatase (ALP) and esterase in HD treatment were lower than LD and MD treatments (P < 0.05) In skin, HD treatment resulted in up-regulation in malondialdehyde (MDA) formation and heat shock protein 70 (HSP 70) mRNA level, and down-regulation in activity of superoxide dismutase (SOD) and the transcriptions of glutathione-s-transferase (GST), interleukin-1ß (IL-1ß), tumor necrosis factor -α (TNF-α), insulin-like growth factor- (IGF-) and LZM (P < 0.05). Overall, the data suggested that overly high stocking density was a stressor which caused an immunosuppression in skin of turbot. Moreover, this information would help to understand the skin immunity and their relation with stress and disease in fish.

Molecular characterization and expression analysis of AMPK α subunit isoform genes from Scophthalmus maximus responding to salinity stress.

AMP-activated protein kinase (AMPK) is a highly conserved and multi-functional protein kinase that plays important roles in both intracellular energy balance and cellular stress response. In the present study, molecular characterization, tissue distribution and gene expression levels of the AMPK α1 and α2 genes from turbot (Scophthalmus maximus) under salinity stress are described. The complete coding regions of the AMPK α1 and α2 genes were isolated from turbot through degenerate primers in combination with RACE using muscle cDNA. The complete coding regions of AMPK α1 (1722 bp) and α2 (1674 bp) encoded 573 and 557 amino acids peptides, respectively. Multiple alignments, structural analysis and phylogenetic tree construction indicated that S. maximus AMPK α1 and α2 shared a high amino acid identity with other species, especially fish. AMPK α1 and α2 genes could be detected in all tested tissues, indicating that they are constitutively expressed. Salinity challenges significantly altered the gene expression levels of AMPK α1 and α2 mRNA in a salinity- and time-dependent manners in S. maximus gill tissues, suggesting that AMPK α1 and α2 played important roles in mediating the salinity stress in S. maximus. The expression levels of AMPK α1 and α2 mRNA were a positive correlation with gill Na+, K+-ATPase activities. These findings will aid our understanding of the molecular mechanism of juvenile turbot in response to environmental salinity changes.

TGF-ß-producing regulatory B cells induce regulatory T cells and promote transplantation tolerance.

Regulatory B (Breg) cells have been shown to play a critical role in immune homeostasis and in autoimmunity models. We have recently demonstrated that combined anti-T cell immunoglobulin domain and mucin domain-1 and anti-CD45RB antibody treatment results in tolerance to full MHC-mismatched islet allografts in mice by generating Breg cells that are necessary for tolerance. Breg cells are antigen-specific and are capable of transferring tolerance to untreated, transplanted animals. Here, we demonstrate that adoptively transferred Breg cells require the presence of regulatory T (Treg) cells to establish tolerance, and that adoptive transfer of Breg cells increases the number of Treg cells. Interaction with Breg cells in vivo induces significantly more Foxp3 expression in CD4(+) CD25(-) T cells than with naive B cells. We also show that Breg cells express the TGF-ß associated latency-associated peptide and that Breg-cell mediated graft prolongation post-adoptive transfer is abrogated by neutralization of TGF-ß activity. Breg cells, like Treg cells, demonstrate preferential expression of both C-C chemokine receptor 6 and CXCR3. Collectively, these findings suggest that in this model of antibody-induced transplantation tolerance, Breg cells promote graft survival by promoting Treg-cell development, possibly via TGF-ß production.

OLC1 is overexpressed in breast cancer and its expression correlates with poor patient survival.

The overexpressed in lung cancer 1 (OLC1) has been demonstrated to be associated with numerous biological and pathological processes. However, the role of OLC1 in breast cancer has not been thoroughly elucidated. The purpose of this study was to assess OLC1 expression and to explore its contribution to the breast cancer. Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) was conducted to detect OLC1 messenger RNA (mRNA) expression in 45 pairs of fresh-frozen breast cancer tissues and corresponding noncancerous tissues. Immunohistochemistry was performed to detect the expression of OLC1 in 145 breast cancer tissues. The relationship between the expression of OLC1 and clinicopathological characteristics and prognosis was statistically analyzed. We found that the expression levels of OLC1 mRNA and protein in breast cancer tissues were significantly higher than those in adjacent noncancerous tissues (P < 0.001). In addition, OLC1 expression was significantly correlated with tumor size (P = 0.034), grade (P = 0.015), stage (P < 0.001), and lymph node metastases (P = 0.028). Kaplan-Meier survival analysis showed that a high expression level of OLC1 resulted in a significantly poor prognosis of breast cancer patients. Further, Cox multivariate analysis indicated that OLC1 expression level was an independent prognostic factor for the overall survival rate of breast cancer patients. These findings provide evidence that a high expression level of OLC1 serves as a biomarker for poor prognosis for breast cancer. Thus, we speculate that OLC1 may be a potential target of antiangiogenic therapy for breast cancer.

An immunosufficient murine model for the study of human islets.

For the sake of therapy of diabetes, it is critical to understand human beta cell function in detail in health and disease. Current studies of human beta cell physiology in vivo are mostly limited to immunodeficient mouse models, which possess significant technical limitations. This study aimed to create a new model for the study of human islets through induction of transplant tolerance in immunosufficient mice. B6 diabetic mice were transplanted with human islets and treated with anti-CD45RB. To assess whether anti-CD45RB-induced transplant tolerance requires B cells, B6 recipients received additional anti-CD20 or B6µMT-/- mice were used. For some anti-CD45RB-treated B6µMT-/- mice, additional anti-CD25 mAb was applied at the early or late stage post-transplant. Immunohistology was performed to show the Foxp3 cells in grafted anti-CD45RB/anti-CD20-treated Foxp3-GFP B6 mice. The results showed that anti-CD45RB alone allowed indefinite graft survival in 26.6% of B6 mice, however 100% of xenografts were accepted in mice treated simultaneously with anti-CD20, and 88.9% of xenografts accepted in anti-CD45RB-treated µMT-/- mice. These µMT-/- mice accepted the islets from another human donor but rejected the islets from baboon. Additional administration of anti-CD25 mAb at the time of transplantation resulted in 100% rejection, whereas 40% of grafts were rejected while the antibody was administrated at days 60 post-transplant. Immunohistologic examination showed Foxp3+ cells accumulated around grafts. We conclude that induction of tolerance to human islets in an immunosufficient mouse model could be generated by targeting murine CD45RB and CD20. This new system will facilitate study of human islets and accelerate the dissection of the critical mechanisms underlying islet health in human disease.

Prediction of bowel necrosis by reduced bowel wall enhancement in closed-loop small bowel obstruction: Quantitative methods.

PURPOSE: To assess diagnostic performance and reproducibility of reduced bowel wall enhancement evaluated by quantitative methods using CT to identify bowel necrosis among closed-loop small bowel obstruction (CL-SBO) patients. METHODS: This retrospective single-center study included patients who diagnosed with CL-SBO caused by adhesion or internal hernia during January 2016 and May 2022. Patients were divided into necrotic group (n = 41) and non-necrotic group (n = 67) according to surgical exploration and postoperative pathology. Two doctors independently measured the attenuation of bowel wall and consensus was reached through panel discussion with a third gastrointestinal radiologist. Reduced bowel wall enhancement was assessed by four quantitative methods. Univariate analyses were used to evaluate the association between each method and bowel necrosis, and kappa/intraclass correlation coefficient values were used to assess interobserver agreement. Diagnostic performance parameters were calculated for each method. RESULTS: Reduced bowel wall enhancement in arterial phase (OR 8.98, P < 0.0001), reduced bowel wall enhancement in portal phase (OR 16.84, P < 0.001), adjusted reduced bowel wall enhancement in arterial phase (OR 29.48, P < 0.001), adjusted reduced bowel wall enhancement in portal phase (OR 145.69, P < 0.001) were significantly associated with bowel necrosis. Adjusted reduced bowel wall enhancement in portal phase had the best diagnostic performance (AUC: 0.92; Youden index: 0.84; specificity: 94.03 %) and interobserver agreement (kappa value of 0.59-0.73) to predict bowel necrosis. CONCLUSION: When assessing reduced bowel enhancement to predict bowel necrosis among CL-SBO patients, using unenhanced CT images and proximal dilated loop as standard references in portal phase is the most accurate quantitative method among those tested.

Manufacturing, quality control, and GLP-grade preclinical study of nebulized allogenic adipose mesenchymal stromal cells-derived extracellular vesicles.

BACKGROUND: Human adipose stromal cells-derived extracellular vesicles (haMSC-EVs) have been shown to alleviate inflammation in acute lung injury (ALI) animal models. However, there are few systemic studies on clinical-grade haMSC-EVs. Our study aimed to investigate the manufacturing, quality control (QC) and preclinical safety of clinical-grade haMSC-EVs. METHODS: haMSC-EVs were isolated from the conditioned medium of human adipose MSCs incubated in 2D containers. Purification was performed by PEG precipitation and differential centrifugation. Characterizations were conducted by nanoparticle tracking analysis, transmission electron microscopy (TEM), Western blotting, nanoflow cytometry analysis, and the TNF-α inhibition ratio of macrophage [after stimulated by lipopolysaccharide (LPS)]. RNA-seq and proteomic analysis with liquid chromatography tandem mass spectrometry (LC-MS/MS) were used to inspect the lot-to-lot consistency of the EV products. Repeated toxicity was evaluated in rats after administration using trace liquid endotracheal nebulizers for 28 days, and respiratory toxicity was evaluated 24 h after the first administration. In vivo therapeutic effects were assessed in an LPS-induced ALI/ acute respiratory distress syndrome (ARDS) rat model. RESULTS: The quality criteria have been standardized. In a stability study, haMSC-EVs were found to remain stable after 6 months of storage at - 80°C, 3 months at - 20 °C, and 6 h at room temperature. The microRNA profile and proteome of haMSC-EVs demonstrated suitable lot-to-lot consistency, further suggesting the stability of the production processes. Intratracheally administered 1.5 × 108 particles/rat/day for four weeks elicited no significant toxicity in rats. In LPS-induced ALI/ARDS model rats, intratracheally administered haMSC-EVs alleviated lung injury, possibly by reducing the serum level of inflammatory factors. CONCLUSION: haMSC-EVs, as an off-shelf drug, have suitable stability and lot-to-lot consistency. Intratracheally administered haMSC-EVs demonstrated excellent safety at the tested dosages in systematic preclinical toxicity studies. Intratracheally administered haMSC-EVs improved the lung function and exerted anti-inflammatory effects on LPS-induced ALI/ARDS model rats.

Mitigation of biofouling with co-deposition of polydopamine and curcumin on the surface of a thin-film composite membrane.

Reverse osmosis (RO) membrane has been widely used in various water treatment fields as an efficient desalination technology, but serious biofouling problem arises in the actual application process. Curcumin is known as a natural compound that can reduce biofouling by inhibiting the growth of microorganisms based on quorum sensing. Dopamine, a molecule with excellent adhesion and functionalization on the material's surface, has high research value for applying a curcumin coating to the membrane surface. Curcumin degrades under alkaline conditions, whereas dopamine must polymerize under alkaline conditions. Simultaneously, a coating may adversely affect curcumin. Therefore, a two-step coating process was considered by self-polymerizing dopamine on the thin-film composite membrane surface and then dip-coating curcumin attached to the polydopamine layer. Furthermore, the effect of time and concentration on the surface modification before and after membrane modification was investigated. The highest permeability of 1.39 L/m2/hr/bar was achieved with the modified membranes. The number of gram-positive bacteria decreased from 6.71 × 106 to 9.67 × 105 CFU/mL. This result is meaningful for antifouling through modification of the membrane surface. Use of curcumin can be applied to reduce biofouling and extend the lifetime of the membrane without pretreatment or membrane cleaning.

Extracellular Non-Coding RNAs in Cardiovascular Diseases.

Cardiovascular diseases (CVDs) remain the world's leading cause of death despite the best available healthcare and therapy. Emerging as a key mediator of intercellular and inter-organ communication in CVD pathogenesis, extracellular vesicles (EVs) are a heterogeneous group of membrane-enclosed nano-sized vesicles released by virtually all cells, of which their RNA cargo, especially non-coding RNAs (ncRNA), has been increasingly recognized as a promising diagnostic and therapeutic target. Recent evidence shows that ncRNAs, such as small ncRNAs, circular RNAs, and long ncRNAs, can be selectively sorted into EVs or other non-vesicular carriers and modulate various biological processes in recipient cells. In this review, we summarize recent advances in the literature regarding the origin, extracellular carrier, and functional mechanisms of extracellular ncRNAs with a focus on small ncRNAs, circular RNAs, and long ncRNAs. The pathophysiological roles of extracellular ncRNAs in various CVDs, including atherosclerosis, ischemic heart diseases, hypertension, cardiac hypertrophy, and heart failure, are extensively discussed. We also provide an update on recent developments and challenges in using extracellular ncRNAs as biomarkers or therapeutical targets in these CVDs.

Finite-element-analysis of connection strength of assembled camshafts with different cam-bore profiles using tube hydroforming technology.

The assembled camshaft is a novel manufacturing product which connects the cam and the mandrel by tube hydroforming (THF) technology after they are processed separately. However, in the process of THF, the structure of the cam-bores has a crucial influence on the connection strength of the assembled camshafts. Therefore, three kinds of cam-bores with circular structure, isometric-trilateral profile and logarithmic spiral profile are selected for hydroforming with a hollow mandrel (tube) in this study. The finite-element-analysis is carried out by ABAQUS software, the variations of (residual) contact pressure and contact area under different structures are obtained, and the torsional angle variations after assembly are measured. Further, the connection strength of the assembled camshaft under three structures is discussed. The results show that the evaluation of connection strength of the assembled camshaft is affected by many factors, including contact pressure, maximum residual contact pressure, axial and circular residual contact pressure, contact area and its rate, residual contact area percentage and torsional angle. Through the comprehensive analysis of various factors, the torsional angle of the camshaft with circular structure is the largest, i.e. poor connection strength. By contrast, the torsional strength of the camshaft with isometric-trilateral profile is the largest, namely, the best connection strength.

Clinical characteristics, treatment patterns and survival outcomes of early-onset pancreatic adenocarcinoma: a population-based study.

BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) is a rare and refractory malignancy. Early-onset pancreatic cancer (EOPC), defined as pancreatic cancer diagnosed before the age of 50 years, is very rare. Clinical presentation and oncological outcomes of EOPC are confusing according to previous studies. METHODS: We performed a retrospective, population-based study by querying the SEER database to analyze patients with PDAC from 2004 to 2018. Data on demographics, pathological characteristics, treatment patterns, and survival outcomes were compared between EOPC and pancreatic cancer in older patients. Propensity score matching (PSM) was used to minimize the potential bias of baseline characteristics between the two groups. The effect of age on changes in treatment modalities was evaluated using the Cochran-Armitage trend test. RESULTS: The entire study enrolled 42,414 patients, including 2,916 (6.9%) patients with EOPC. Patients with EOPC were more likely to be male (56.6% vs. 51.0%, P < 0.001) and more frequently to present with a larger tumor size (40 mm vs. 37 mm, P < 0.001), vascular invasion (28.6% vs. 25.9%, P = 0.022) and distant metastasis (56.2% vs. 50.8%, P < 0.001) compared with older group. However, surgical resection rates (29.3% vs. 28.3%, P = 0.284) were fairly comparable, and most clinicopathologic characteristics were similar in the patients underwent resection. Younger patients had longer 5-year overall survival (6.9% vs. 5.5%, P < 0.001) and 5-year cancer-specific survival (8.4% vs. 7.3%, P < 0.001) among the overall cohort but had comparable prognosis among patients received surgery (both P > 0.05). Similar survival outcomes were obtained after PSM. In addition, operated patients tended to receive fewer systemic treatments at an increasing age (Ptrend < 0.001). The survival analysis, which was stratified by age groups, suggested that younger patients only had a better prognosis than those over 70. CONCLUSIONS: Patients with EOPC exhibited an advanced stage and a male predilection at diagnosis in the overall cohort but broadly similar clinicopathologic characteristics in the operated patients. In the surgical cohort, although younger patients were more likely to receive systemic treatment, patients with EOPC presented comparable outcomes compared with elderly patients. We suggest that more research should be conducted to uncover the unique characteristics of EOPC for better clinical management.

Preventive therapeutic effect of Lactobacillus-fermented black wolfberry juice on sodium dextran sulfate-induced ulcerative colitis in mice.

In this study, male mice were treated with fermented and unfermented Lactobacillus plantarum, Lactobacillus bulgaricus, and Lactobacillus rhamnosus black wolfberry juice (10 mL/kg/day) for 40 days, and their prophylactic effects on ulcerative colitis (UC) induced by dextran sodium sulfate were investigated. The intervention of black wolfberry juice reduced the levels of pro-inflammatory cytokines and increased the content of anti-inflammatory cytokines in the serum and colon. In addition, the pathological changes in colon tissue were alleviated, the expression of Bcl-2 protein in the colon was enhanced, and the intestinal microbiota of the mice was regulated, with an increase in Bacteroidetes and a decrease in Helicobacter. These results suggested that black wolfberry juice had an anti-UC function and Lactobacillus fermentation enhanced the anti-inflammatory effect of black wolfberry juice by modulating the intestinal microbiota.

Experience in the management of sigmoid sinus thrombophlebitis secondary to middle ear cholesteatoma.

OBJECTIVE: To discuss the management of sigmoid sinus thrombophlebitis secondary to middle ear cholesteatoma. METHODS: We retrospectively analyzed all cases of sigmoid sinus thrombophlebitis caused by middle ear cholesteatoma over a period of 7 years. 7 male and 2 female patients, ranging in age from 9 to 66 years, were diagnosed with sigmoid sinus thrombophlebitis by clinical presentation and radiological examination. By executing a modified mastoidectomy and tympanoplasty (canal wall-down tympanoplasty) to entirely remove the cholesteatoma-like mastoid epithelium, all patients were effectively treated surgically without opening the sigmoid sinus. All patients were treated with broad-spectrum antibiotics, but no anticoagulants were used. RESULTS: 9 patients had otogenic symptoms such as ear pus, tympanic membrane perforation, and hearing loss. In the initial stage of the surgery, modified mastoidectomy and tympanoplasty were performed on 8 of the 9 patients. 1 patient with a brain abscess underwent puncturing (drainage of the abscess) to relieve cranial pressure, and 4 months later, a modified mastoidectomy and tympanoplasty were carried out. Following surgery and medication, the clinical symptoms of every patient improved. After the follow-up of 6 months to 7 years, 3 patients were re-examined for MRV and showed partial sigmoid sinus recovery with recanalization. 4 months following middle ear surgery, the extent of a patient's brain abscess lesions was significantly reduced. 1 patient experienced facial paralysis after surgery and recovered in 3 months. None of the patients had a secondary illness, an infection, or an abscess in a distant organ. CONCLUSION: The key to a better prognosis is an adequate course of perioperative antibiotic medication coupled with surgical treatment. A stable sigmoid sinus thrombus can remain for a long time after middle ear lesions have been removed, and it is less likely to cause infection and abscesses in the distant organs. The restoration of middle ear ventilation is facilitated by tympanoplasty. It is important to work more closely with multidisciplinary teams such as neurology and neurosurgery when deciding whether to perform lateral sinusotomies to remove thrombus or whether to administer anticoagulation.