3D printing-based full-scale human brain for diverse applications.

Surgery is the most frequent treatment for patients with brain tumors. The construction of full-scale human brain models, which is still challenging to realize via current manufacturing techniques, can effectively train surgeons before brain tumor surgeries. This paper aims to develop a set of three-dimensional (3D) printing approaches to fabricate customized full-scale human brain models for surgery training as well as specialized brain patches for wound healing after surgery. First, a brain patch designed to fit a wound's shape and size can be easily printed in and collected from a stimuli-responsive yield-stress support bath. Then, an inverse 3D printing strategy, called "peeling-boiled-eggs," is proposed to fabricate full-scale human brain models. In this strategy, the contour layer of a brain model is printed using a sacrificial ink to envelop the target brain core within a photocurable yield-stress support bath. After crosslinking the contour layer, the as-printed model can be harvested from the bath to photo crosslink the brain core, which can be eventually released by liquefying the contour layer. Both the brain patch and full-scale human brain model are successfully printed to mimic the scenario of wound healing after removing a brain tumor, validating the effectiveness of the proposed 3D printing approaches.

Intracranial Rosai­Dorfman disease complicated by mucosa­associated lymphoid tissue: A case report.

Intracranial Rosai-Dorfman disease (RDD) is a rare, self-limiting histiocytic disease of unknown etiology. Extranodal marginal zone lymphoma of the mucosa-associated lymphoid tissue (MALT lymphoma) is also rare and intracranial RDD complicated by MALT lymphoma is even rarer. The present study reports a case of a 55-year-old female who was admitted to The Second Affiliated Hospital of Jiaxing University (Jiaxing, China) with headache for half a month and ptosis of the right eyelid for 4 days. Computerised tomography and magnetic resonance imaging revealed a right parasellar tumor and, subsequently, subtotal resection of the tumor was performed. Postoperative pathology revealed intracranial RDD complicated by MALT lymphoma. The patient received chemotherapy after surgery and achieved good therapeutic effects. After 12 months of follow-up, the residual tumor disappeared and the ptosis prominently improved. To the to the best of the authors' knowledge, the present case is the first reported case of an adult intracranial RDD complicated by MALT lymphoma.

Cedrol, a Ginger-derived sesquiterpineol, suppresses estrogen-deficient osteoporosis by intervening NFATc1 and reactive oxygen species.

Osteoporosis is a prevalent bone metabolic disease in menopause, and long-term medication is accompanied by serious side effects. Ginger, a food spice and traditional medicine with ancient history, exhibits the potential to alleviate osteoporosis in preclinical experiments, whereas its complex composition leads to ambiguous pharmacological mechanisms. The purpose of this study was to investigate the effect and mechanism of Ced in estrogen-deficient osteoporosis, a sesquiterpene alcohol recently discovered from Ginger with multiple pharmacological properties. RANKL was stimulated BMM (bone marrow macrophages) differentiation into osteoclasts in vitro. And the osteoclast activity and number were assessed by TRAcP and SEM. We found that Ced mitigated RANKL-induced osteoclastogenesis by descending the ROS content and obstructing NFATc1, NF-κB, and MAPK signaling. Also, Ced-mediated anti-osteolytic property was found in ovariectomized mice by Micro-CT scanning and histological staining. Summarily, our works demonstrated the anti-osteoporotic potential of Cedrol in Ginger for the first time, which also offered more pharmacological evidence for Ginger as food or medicine used for bone metabolic disease.

Olfactory receptor 2 in vascular macrophages drives atherosclerosis by NLRP3-dependent IL-1 production.

Atherosclerosis is an inflammatory disease of the artery walls and involves immune cells such as macrophages. Olfactory receptors (OLFRs) are G protein­coupled chemoreceptors that have a central role in detecting odorants and the sense of smell. We found that mouse vascular macrophages express the olfactory receptor Olfr2 and all associated trafficking and signaling molecules. Olfr2 detects the compound octanal, which activates the NLR family pyrin domain containing 3 (NLRP3) inflammasome and induces interleukin-1ß secretion in human and mouse macrophages. We found that human and mouse blood plasma contains octanal, a product of lipid peroxidation, at concentrations sufficient to activate Olfr2 and the human ortholog olfactory receptor 6A2 (OR6A2). Boosting octanal levels exacerbated atherosclerosis, whereas genetic targeting of Olfr2 in mice significantly reduced atherosclerotic plaques. Our findings suggest that inhibiting OR6A2 may provide a promising strategy to prevent and treat atherosclerosis.

Bone Marrow Transplantation Rescues Monocyte Recruitment Defect and Improves Cystic Fibrosis in Mice.

Cystic fibrosis (CF) is an inherited life-threatening disease accompanied by repeated lung infections and multiorgan inflammation that affects tens of thousands of people worldwide. The causative gene, cystic fibrosis transmembrane conductance regulator (CFTR), is mutated in CF patients. CFTR functions in epithelial cells have traditionally been thought to cause the disease symptoms. Recent work has shown an additional defect: monocytes from CF patients show a deficiency in integrin activation and adhesion. Because monocytes play critical roles in controlling infections, defective monocyte function may contribute to CF progression. In this study, we demonstrate that monocytes from CFTRΔF508 mice (CF mice) show defective adhesion under flow. Transplanting CF mice with wild-type (WT) bone marrow after sublethal irradiation replaced most (60-80%) CF monocytes with WT monocytes, significantly improved survival, and reduced inflammation. WT/CF mixed bone marrow chimeras directly demonstrated defective CF monocyte recruitment to the bronchoalveolar lavage and the intestinal lamina propria in vivo. WT mice reconstituted with CF bone marrow also show lethality, suggesting that the CF defect in monocytes is not only necessary but also sufficient to cause disease. We also show that monocyte-specific knockout of CFTR retards weight gains and exacerbates dextran sulfate sodium-induced colitis. Our findings show that providing WT monocytes by bone marrow transfer rescues mortality in CF mice, suggesting that similar approaches may mitigate disease in CF patients.

Identification of water-soluble peptides in distilled spent grain and its angiotensin converting enzyme (ACE) inhibitory activity based on UPLC-Q-TOF-MS and proteomics analysis.

Distilled spent grain (DSG) is the biggest by-product in baijiu (Chinese liquor) production, releasing approximately 23.44 million tons every year. Aiming at comprehensive identification of more bioactive peptides, in this work, the new bioassay-guided proteomics and Biolynx peptide sequencer based on ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) were developed. Moreover, 22 peptides with angiotensin converting enzyme (ACE) inhibitory activities were identified. Seven peptides were successfully quantified using electrospray ionization with triple-quadrupole mass spectrometry (ESI-QQQ-MS) in the multiple reaction monitoring (MRM). Of these identified peptides, Pro-Arg was the most abundant (92.14 µg g-1 dry weight (DW)) and acted as a competitive inhibitor of ACE by molecular docking. Therefore, peptides from DSG can be considered as promising candidates for ACE inhibition; in addition, the new strategy for peptide sequencing can be extended to any food matrices containing peptide mixture or protein hydrolysate.

Kindlin-3 recruitment to the plasma membrane precedes high-affinity ß2-integrin and neutrophil arrest from rolling.

Integrin-mediated neutrophil adhesion starts by arrest from rolling. Activation of integrins involves conformational changes from an inactive, bent conformation to an extended conformation (E+) with high affinity for ligand binding (H+). The cytoplasmic protein kindlin-3 is necessary for leukocyte adhesion; mutations of kindlin-3 cause leukocyte adhesion deficiency type 3. Kindlin-3 binds the ß2-integrin cytoplasmic tail at a site distinct from talin-1, but the molecular mechanism by which kindlin-3 activates ß2-integrins is unknown. In this study, we measured the spatiotemporal dynamics of kindlin-3 and ß2-integrin conformation changes during neutrophil and HL-60 cell rolling and arrest under flow. Using high-resolution quantitative dynamic footprinting microscopy and kindlin-3-fluorescent protein (FP) fusion proteins, we found that kindlin-3 was recruited to the plasma membrane in response to interleukin-8 (IL-8) before induction of the H+ ß2-integrin conformation. Intravital imaging revealed that EGFP-kindlin-3-reconstituted, kindlin-3-knockout neutrophils arrest in vivo in response to CXCL1. EGFP-kindlin-3 in primary mouse neutrophils was also recruited to the plasma membrane before arrest. Upon arrest, we found small clusters of high-affinity ß2-integrin molecules within large areas of membrane-proximal kindlin-3 FP. Deletion of kindlin-3 or its pleckstrin homology (PH) domain in neutrophil-like HL-60 cells completely abolished H+ ß2-integrin induction. IL-8 also triggered recruitment of the isolated kindlin-3 PH domain to the plasma membrane before arrest. In summary, we showed that the kindlin-3 PH domain is necessary for recruitment to the plasma membrane, where full-length kindlin-3 is indispensable for the induction of high-affinity ß2-integrin.

Hepatic portal venous gas: A case report and analysis of 131 patients using PUBMED and MEDLINE database.

OBJECTIVE: Hepatic portal pneumatosis has a high mortality rate, and whether surgical intervention is necessary remains controversial. This experiment retrospectively analyzed the etiology, treatment methods and prognosis of adult patients with hepatoportal pneumocele to provide a theoretical basis for the treatment of this disease. METHODS: We analyzed the clinical symptoms and post-treatment of a 43-year-old male patient with HPVG admitted to hospital. We retrieved adult non-iatrogenic HPVG cases with complete clinical data in PUBMED,  and MEDLINE and other databases were retrieved for analysis, and summarized the pathogenesis, clinical symptoms, pathogenesis, pathogenesis and prognosis of different treatment schemes were summarized. RESULTS: The main etiology of HPVG are intestinal ischemia (27%), severe enteritis/intestinal perforation/intestinal fistula (16%), intestinal obstruction (7%), abdominal infection (7%), gastric diseases (11%), appendicitis and its complications (5%), acute hemorrhage or necrotizing pancreatitis (5%), Crohn's disease and its complications (4%), trauma (traffic accidents, falls) (2%), diverticulitis and perforation (6%), nephrogenic diseases (4%), spontaneous pneumohepatic portal vein (2%), other reasons (4%). And after analysis, we found that the survival rate of patients treated by surgery was 40.5% and the mortality rate was 19.1%, the difference between the two was significant. CONCLUSIONS: Etiology should be actively explored and surgical treatment is necessary.

Localised delivery of quercetin by thermo-sensitive PLGA-PEG-PLGA hydrogels for the treatment of brachial plexus avulsion.

Accumulating evidence indicates that oxidative stress and inflammation are implicated in brachial plexus avulsion (BPA). Quercetin has anti-inflammatory, anti-oxidant, anti-apoptotic, and neuroprotective properties. This study investigated the therapeutic efficacy of a temperature-sensitive poly(D,L-lactide-co-glycolide)-poly(ethylene-glycol)-poly(D,L-lactide-co-glycolide) (PLGA-PEG-PLGA) hydrogel sustained-release system of quercetin in BPA. In situ injections of the hydrogel loaded with different concentrations of quercetin were conducted in a rat model of BPA. Significantly reduced reactive oxygen species and interleukin-6 levels in the injured spinal cord 24 h post-surgery, increased number of anterior horn motor and functional neurons in the spinal cord 6 weeks post-surgery, thickened biceps muscle fibres and enlarged endplate area with clear structure, reduced demyelinated peripheral nerves, and significantly increased Terzis grooming test scores were found in the groups with 50 or 100 mg/mL quercetin-loaded hydrogels compared with the control and blank hydrogel groups. In conclusion, the temperature-sensitive quercetin loaded PLGA-PEG-PLGA hydrogel sustained-release system can alleviate oxidative damage and inflammation in the spinal cord, increase neuron survival rate, and promote nerve regeneration and motor function recovery in rats with early BPA. The findings suggest that this drug-loaded hydrogel has potential applications in the clinical treatment of BPA.

Imaging of the immune system - towards a subcellular and molecular understanding.

Immune responses involve many types of leukocytes that traffic to the site of injury, recognize the insult and respond appropriately. Imaging of the immune system involves a set of methods and analytical tools that are used to visualize immune responses at the cellular and molecular level as they occur in real time. We will review recent and emerging technological advances in optical imaging, and their application to understanding the molecular and cellular responses of neutrophils, macrophages and lymphocytes. Optical live-cell imaging provides deep mechanistic insights at the molecular, cellular, tissue and organism levels. Live-cell imaging can capture quantitative information in real time at subcellular resolution with minimal phototoxicity and repeatedly in the same living cells or in accessible tissues of the living organism. Advanced FRET probes allow tracking signaling events in live cells. Light-sheet microscopy allows for deeper tissue penetration in optically clear samples, enriching our understanding of the higher-level organization of the immune response. Super-resolution microscopy offers insights into compartmentalized signaling at a resolution beyond the diffraction limit, approaching single-molecule resolution. This Review provides a current perspective on live-cell imaging in vitro and in vivo with a focus on the assessment of the immune system.

l-Theanine and NEP1-40 promote nerve regeneration and functional recovery after brachial plexus root avulsion.

Brachial plexus root avulsion causes severe sequelae Treatments and prognosis face many problems, including inflammatory reaction, oxidative damage, and myelin related inhibitory effect. l-Theanine has anti-inflammatory, anti-oxidative, and neuroprotective effects. NEP1-40 competitively inhibits Nogo-66 receptor (NgR1) promotes axonal regeneration. Forty-eight Sprague-Dawley rats were randomly assigned into four groups to establish an animal model of brachial plexus root avulsion. Inflammation and oxidative damage were evaluated by spectrophotometry and motor function of the upper limbs was assessed via Terzis grooming test after modeling. Immunofluorescence and hematoxylin and eosin staining were utilized to determine the content of reactive oxygen species, activation of microglial cells, neuroprotection, and nerve regeneration. Compared with the control group, the L-Theanine + NEP1-40 group had significantly decreased myeloperoxidase, malondialdehyde, interleukin-6, reactive oxygen species, and microglial cells, significantly increased score on the Terzis grooming test, increased motor neuron content, and thickened muscle fibers, increased area, and appearance of large and clear motor endplate structures. The results of this study suggest that l-Theanine combined with NEP1-40significantly promoted nerve regeneration after brachial plexus root avulsion, and may be a potential treatment for promoting nerve regeneration. Possible mechanisms underlying these results are alleviation of oxidative damage and inflammatory responses in the injured area and antagonism of myelin inhibition.

Analgesic therapy improves arterial endothelial function following non-cardiovascular surgery: A randomized, placebo-controlled trial.

Pain subsequent to non-cardiac surgery may affect the endothelial function, which in turn contributes to myocardial injury (MI). The present study examined whether effective pain control is able to improve the postoperative endothelial function. Patients (n=160) undergoing laparoscopic cholecystectomy were randomly assigned into two groups, treated with tramadol analgesic or saline (placebo) following surgery. On preoperative day 1 (baseline) and postoperatively at 2 h, 1 day and 5 days, pain was assessed on a visual analogue scale (VAS), and B-mode ultrasound was used to measure brachial endothelium-dependent flow-mediated dilation (FMD) and nitroglycerin-induced dilation. At 2 h postoperatively, the FMD in the two groups was significantly lower compared with that at the other three time points (P≤0.005), while VAS was significantly higher (P<0.05). Patients in the tramadol group presented significantly reduced VAS values in comparison with those in the placebo group at 2 h and 1 day postoperatively (P=0.013 and 0.031, respectively), as well as significantly higher FMD at 2 h (6.7±1.5 vs. 6.0±1.7%; P=0.001) and 1 day postoperatively (7.3±1.3 vs. 6.9±1.4%; P=0.03). A VAS score of <5 was independently associated with postoperative FMD of ≥7 (odds ratio, 2.5; 95% confidence interval, 1.0-6.0; P=0.047). Backward multivariate linear regression also demonstrated that FMD was independently correlated with age and VAS score (B=-1.403, P=0.011; B=-0.579, P=0.003). The response to nitroglycerin-induced dilation remained stable in all patients at baseline and at all postoperative time points. In conclusion, analgesic treatment may improve the arterial endothelial function following non-cardiac surgery, which may help prevent postoperative MI.

Preoperative and postoperative analgesic techniques in the treatment of patients undergoing transabdominal hysterectomy: a preliminary randomized trial.

BACKGROUND: Although pre-emptive analgesia is commonly used for the management of postoperative pain in developed countries, no defined protocol has been carried out and widely practiced, especially in transabdominal hysterectomy. Keeping this in mind the present study aimed to investigate the effects of multimodal pre-emptive analgesia on pain management, stress response and inflammatory factors of patients undergoing transabdominal hysterectomy to find an optimized way of pre-emptive analgesia. METHODS: One hundred patients undergoing abdominal hysterectomy were randomly divided into four groups (Trial registration: ChiCTR-IPR-15005848). Group P1 was given intravenous flurbiprofen and epidural fentanyl + ketamine before surgery; Group P2 received intravenous flurbiprofen before surgery and epidural fentanyl + ketamine after surgery; Group P3 was given epidural fentanyl + ketamine before surgery and intravenous flurbiprofen after surgery; Patients in Group C received normal saline treatment. RESULTS: Compared with control group, the first time to request additional analgesics after surgery were significantly later (P < 0.05), 24 h dosage of analgesia were significantly less (P < 0.05), VAS score at all time periods after surgery were significantly lower (P < 0.05) in Group P1, P2, or P3. At 12 h or 24 h after surgery, VAS score in Group P1 was significantly lower than that in group P2 or P3 (P < 0.05, P < 0.05). No significant adverse effects were found among the groups (P > 0.05). At 1 or 2 days after surgery, the levels of cortisol, glucose, and IL-6, TNF-α in group P1, P2, and P3 were significantly lower than those in group C (P < 0.05); while, the levels in group P2, P3 were significantly lower than those in group P1 (P < 0.05). CONCLUSION: Multimodal pre-emptive analgesia could significantly lower VAS score, inhibit stress response, and reduce inflammatory response in patients undergoing transabdominal hysterectomy, which can be a rational strategy for pain control in future. TRIAL REGISTRATION: ChiCTR-IPR-15005848 on January 17, 2015.

Therapeutic effect of aqueous extract from Ecliptae herba on bone metabolism of ovariectomized rats.

OBJECTIVE: Ecliptae herba (EH) has long been used in China to strengthen bones. Accumulating evidence indicates that EH may have antiosteoporotic effects. The aim of this study was to evaluate the effects of aqueous EH extract (EHE) on rats that had osteoporosis-like features induced by ovariectomy, using aqueous Fructus Ligustri Lucidi extract as positive control agent. METHODS: Three-month-old female rats that underwent ovariectomy were treated with EHE (1.4 g/kg per day). After 12 weeks, bone mineral density and bone histomorphometric indices of tibiae were measured. Protein and messenger RNA expressions of osteoprotegerin and receptor activator of nuclear factor κ-B ligand (RANKL) in tibiae were evaluated by immunohistochemistry and in situ hybridization. In addition, serum concentrations of osteocalcin, interleukin-1ß, interleukin-6 (IL-6), calcitonin (CT), and parathyroid hormone were determined by enzyme-linked immunosorbent assay. RESULTS: EHE treatment prevented body weight gain and loss of uterine wet weight in ovariectomized rats. It remarkably increased bone mass in ovariectomized rats compared with ovariectomized controls. EHE treatment significantly down-regulated RANKL expression in tibiae from ovariectomized rats compared with controls; however, it had no significant effect on osteoprotegerin expression. In addition, EHE treatment significantly reduced serum IL-6 levels and remarkably increased CT levels but had no effect on parathyroid hormone. CONCLUSIONS: EHE increases bone mass in ovariectomized rats by inhibiting bone loss: down-regulated RANKL expression in tibiae and IL-6 level in serum, and up-regulated CT level in serum. This suggests that EHE may be developed as an alternative therapeutic agent for osteoporosis induced by postmenopause.

Isolation of mouse mesenchymal stem cells with normal ploidy from bone marrows by reducing oxidative stress in combination with extracellular matrix.

BACKGROUND: Isolation of mouse MSCs (mMSCs) with normal ploidy from bone marrow remains challenging. mMSCs isolated under 20% O(2) are frequently contaminated by overgrown hematopoietic cells, and could also be especially vulnerable to oxidative damage, resulting in chromosomal instability. Culture under low oxygen or extracellular matrix (ECM) improves proliferation of MSCs in several species. We tested the hypothesis that culture under low oxygen in combination with ECM prepared from mouse embryonic fibroblast (MEF-ECM) could be used to purify proliferative mMSCs, and to reduce oxidative damage and maintain their chromosomal stability. RESULTS: Optimization of culture conditions under 20% O(2) resulted in immortalization of mMSCs, showing extensive chromosome abnormalities, consistent with previous studies. In contrast, culture under low oxygen (2% O(2)) improved proliferation of mMSCs and reduced oxidative damage, such that mMSCs were purified simply by plating at low density under 2% O(2). MEF-ECM reduced oxidative damage and enhanced proliferation of mMSCs. However, these isolated mMSCs still exhibited high frequency of chromosome abnormalities, suggesting that low oxygen or in combination with MEF-ECM was insufficient to fully protect mMSCs from oxidative damage. Notably, antioxidants (alpha -phenyl-t-butyl nitrone (PBN) and N-acetylcysteine (NAC)) further reduced DNA damage and chromosomal abnormalities, and increased proliferation of mMSCs. mMSCs isolated by the combination method were successfully used to generate induced pluripotent stem (iPS) cells by ectopic expression of Oct4, Sox2, Klf4 and c-Myc. CONCLUSIONS: We have developed a technique that allows to reduce the number of karyotypic abnormalities for isolation of primary mMSCs and for limited culture period by combination of low oxygen, MEF-ECM, antioxidants and low density plating strategy. The effectiveness of the new combination method is demonstrated by successful generation of iPS cells from the isolated mMSCs. However, a culture system for mMSCs still is needed to prevent all the anomalies, especially after a long-term culture period.