UBE2C orchestrates bone formation through stabilization of SMAD1/5.

While previous studies have demonstrated the role of ubiquitin-conjugating enzyme 2C (UBE2C) in promoting ß-cell proliferation and cancer cell lineage expansion, its specific function and mechanism in bone marrow mesenchymal stem/stromal cells (BMSCs) growth and differentiation remain poorly understood. Our findings indicate that mice with conditional Ube2c deletions in BMSCs and osteoblasts exhibit reduced skeletal bone mass and impaired bone repair. A significant reduction in the proliferative capacity of BMSCs was observed in conditional Ube2c knockout mice, with no effect on apoptosis. Additionally, conditional Ube2c knockout mice exhibited enhanced osteoclastic activity and reduced osteogenic differentiation. Furthermore, human BMSCs with stable UBE2C knockdown exhibited diminished capacity for osteogenic differentiation. Mechanistically, we discovered that UBE2C binds to and stabilizes SMAD1/5 protein expression levels. Interestingly, UBE2C's role in regulating osteogenic differentiation and SMAD1/5 expression levels appears to be independent of its enzymatic activity. Notably, UBE2C regulates osteogenic differentiation through SMAD1/5 signaling. In conclusion, our findings underscore the pivotal role of UBE2C in bone formation, emphasizing its contribution to enhanced osteogenic differentiation through the stabilization of SMAD1/5. These results propose UBE2C as a promising target for BMSC-based bone regeneration.

M2 Macrophages Guide Periosteal Stromal Cell Recruitment and Initiate Bone Injury Regeneration.

The periosteum plays a critical role in bone repair and is significantly influenced by the surrounding immune microenvironment. In this study, we employed 10× single-cell RNA sequencing to create a detailed cellular atlas of the swine cranial periosteum, highlighting the cellular dynamics and interactions essential for cranial bone injury repair. We noted that such injuries lead to an increase in M2 macrophages, which are key in modulating the periosteum's immune response and driving the bone regeneration process. These macrophages actively recruit periosteal stromal cells (PSCs) by secreting Neuregulin 1 (NRG1), a crucial factor in initiating bone regeneration. This recruitment process emphasizes the critical role of PSCs in effective bone repair, positioning them as primary targets for therapeutic interventions. Our results indicate that enhancing the interaction between M2 macrophages and PSCs could significantly improve the outcomes of treatments aimed at cranial bone repair and regeneration.

General Approach to Amides through Decarboxylative Radical Cross-Coupling of Carboxylic Acids and Isocyanides.

Herein, we report a silver-catalyzed protocol for decarboxylative cross-coupling between carboxylic acids and isocyanides, leading to linear amide products through a free-radical mechanism. The disclosed approach provides a general entry to a variety of decorated amides, accommodating a diverse array of radical precursors, including aryl, heteroaryl, alkynyl, alkenyl, and alkyl carboxylic acids. Notably, the protocol proved to be efficient for decarboxylative late-stage functionalization of several elaborate pharmaceuticals, demonstrating its potential applications.

The expression mechanism of programmed cell death 1 ligand 1 and its role in immunomodulatory ability of mesenchymal stem cells.

Programmed cell death 1 ligand 1 (PD-L1) is an important immunosuppressive molecule, which inhibits the function of T cells and other immune cells by binding to the receptor programmed cell death-1. The PD-L1 expression disorder plays an important role in the occurrence, development, and treatment of sepsis or other inflammatory diseases, and has become an important target for the treatment of these diseases. Mesenchymal stem cells (MSCs) are a kind of pluripotent stem cells with multiple differentiation potential. In recent years, MSCs have been found to have a strong immunosuppressive ability and are used to treat various inflammatory insults caused by hyperimmune diseases. Moreover, PD-L1 is deeply involved in the immunosuppressive events of MSCs and plays an important role in the treatment of various diseases. In this review, we will summarize the main regulatory mechanism of PD-L1 expression, and discuss various biological functions of PD-L1 in the immune regulation of MSCs.

The expression mechanism of programmed cell death 1 ligand 1 and its role in immunomodulatory ability of mesenchymal stem cells / 中华创伤杂志(英文版)

Programmed cell death 1 ligand 1 (PD-L1) is an important immunosuppressive molecule, which inhibits the function of T cells and other immune cells by binding to the receptor programmed cell death-1. The PD-L1 expression disorder plays an important role in the occurrence, development, and treatment of sepsis or other inflammatory diseases, and has become an important target for the treatment of these diseases. Mesenchymal stem cells (MSCs) are a kind of pluripotent stem cells with multiple differentiation potential. In recent years, MSCs have been found to have a strong immunosuppressive ability and are used to treat various inflammatory insults caused by hyperimmune diseases. Moreover, PD-L1 is deeply involved in the immunosuppressive events of MSCs and plays an important role in the treatment of various diseases. In this review, we will summarize the main regulatory mechanism of PD-L1 expression, and discuss various biological functions of PD-L1 in the immune regulation of MSCs.

Design, synthesis, and biological evaluation of harmine derivatives as topoisomerase I inhibitors for cancer treatment.

A series of ß-carboline derivatives were designed and synthesized by introducing the chalcone moiety into the harmine. The synthesized derivatives were evaluated their anti-proliferative activities against six human cancer cell lines (MCF-7, MDA-MB-231, HepG2, HT29, A549, and PC-3) and one normal cell line (L02). Among them, compound G11 exhibited the potent anti-proliferative activity against MCF-7 cell line, with an IC50 value of 0.34 µM. Further biological studies revealed that compound G11 inhibited colony formation of MCF-7 cells, suppressed MCF-7 cell migration by downregulating migration-associated protein MMP-2. In addition, it could induce apoptosis of MCF-7 cells by downregulating Bcl-2 and upregulating Cleaved-PARP, Bax, and phosphorylated Bim proteins. Furthermore, compound G11 can act as a Topo I inhibitor, affecting DNA synthesis and transcription, thereby inhibiting cancer cell proliferation. Moreover, compound G11 inhibited tumor growth in 4T1 syngeneic transplant mice with an inhibition rate of 43.19 % at a dose of 10 mg/kg, and 63.87 % at 20 mg/kg, without causing significant toxicity to the mice or their organs, achieving the goal of reduced toxicity and increased efficacy. All these results indicate of G11 has enormous potential as an anti-tumor agent and merits further investigation.

Meroterpenoids with divers' rings systems from Phyllosticta capitalensis and their anti-inflammatory activity.

Four undescribed sesquiterpene-shikimates (1-4), eight undescribed monoterpene-shikimates (5-12), together with two known ones were isolated and identified from the 95% ethanol extract of the plant endophytic fungus Phyllosticta capitalensis cultured in rice medium. Capitalensis A (1) was identified as the first sesquiterpene-shikimate-conjugated spirocyclic meroterpenoid degradation product, while capitalensis B (2) is a sesquiterpene-shikimate-conjugated spirocyclic meroterpenoid with a unique D-ring formed by a C-2-O-C-9' connection. The structures of these previously undescribed compounds were elucidated by multiple techniques, including IR, HR-ESI-MS, and NMR analysis. Furthermore, their absolute configurations were established through the comprehensive approach that involved the calculations of ECD spectra, optical rotation values, and single-crystal X-ray analysis. Moreover, the anti-inflammatory activity of all isolated compounds was evaluated using a lipopolysaccharide (LPS)-induced inflammation model in BV2 microglial cells. Meanwhile, these compounds exhibited activity in inhibiting NO production. Four compounds, capitalensis C (3), capitalensis D (4), 15-hydroxyl tricycloalternarene 5b (13) and guignarenone A (14) showed strong inhibitory effects with IC50 values of 21.6 ± 1.33, 12.2 ± 1.08, 18.6 ± 1.27, and 15.8 ± 1.20 µM, respectively. In addition, the structure-activity relationship of the anti-inflammatory activity of the compounds was discussed.

Organoid Culture Development for Skeletal Systems.

Organoids are widely considered to be ideal in vitro models that have been widely applied in many fields, including regenerative medicine, disease research and drug screening. It is distinguished from other three-dimensional in vitro culture model systems by self-organization and sustainability in long-term culture. The three core components of organoid culture are cells, exogenous factors, and culture matrix. Due to the complexity of bone tissue, and heterogeneity of osteogenic stem/progenitor cells, it is challenging to construct organoids for modeling skeletal systems. In this study, we examine current progress in the development of skeletal system organoid culture systems and analyze the current research status of skeletal stem cells, their microenvironmental factors, and various potential organoid culture matrix candidates to provide cues for future research trajectory in this field. Impact Statement The emergence of organoids has brought new opportunities for the development of many biomedical fields. The bone organoid field still has much room for exploration. This review discusses the characteristics distinguishing organoids from other three-dimensional model systems and examines current progress in the organoid production of skeletal systems. In addition, based on core elements of organoid cultures, three main problems that need to be solved in bone organoid generation are further analyzed. These include the heterogeneity of skeletal stem cells, their microenvironmental factors, and potential organoid culture matrix candidates. This information provides direction for the future research of bone organoids.

The value of CT radiomics in predicting severe hyperlipidemic acute pancreatitis / 实用放射学杂志

Objective To explore the value of CT enhanced radiomics model in predicting severe hyperlipidemic acute pancreatitis(HLAP).Methods The data of 117 HLAP patients were analyzed retrospectively and the patients were randomly divided into a training set(93 cases)and a test set(24 cases)in the ratio of 8∶2.CT enhanced images of arterial phase and venous phase were collected,and the optimal radiomics features were extracted and screened.The arterial phase model and venous phase model were established by support vector machine(SVM).Meanwhile,the bedside index for severity in acute pancreatitis(BISAP)was scored and a BISAP model was developed for the patients based on clinical information.The area under the curve(AUC)under the receiver operating characteristic(ROC)curve was used as the evaluation criterion for the model.DeLong test was used to compare the prediction efficiency of each model.Results The training set AUC of the arterial phase model,venous phase model and BISAP model were 0.777,0.788 and 0.732,respectively.And the AUC of the test set were 0.836,0.734 and 0.695,respectively.DeLong test results showed that the training set AUC of the arterial phase model and the venous phase model was better than that of BISAP model(P＜0.05),but there was no significant difference between the AUC of the test set AUC(P＞0.05).There was no significant difference in AUC between arterial phase model and venous phase model(P＞0.05).Conclusion The radiomics model based on CT enhancement can predict the severity of HLAP at the early stage,which helps to target the treatment of HLAP patients in the early clinical stage.

Study on the mechanism of Chonghe paste promoting the dissipation of swollen lesions in vivo and in vitro / 中国药房

OBJECTIVE To explore the mechanism of Chonghe paste promoting the dissipation of swollen lesions. METHODS The bacteriostatic effects of Chonghe paste against Staphylococcus aureus， Escherichia coli， Pseudomonas aeruginosa， Staphylococcus albus and Streptococcus pneumoniae were detected by punching method. The subcutaneous soft tissue infection model of rats was established by subcutaneous injection of S. aureus. The effects of 14 d intervention of Chonghe paste （Compound polymyxin B ointment as positive control） on the pathological changes of subcutaneous soft tissue， the protein expressions of type Ⅰ collagen， type Ⅲ collagen， matrix metalloproteinase-2 （MMP-2） and MMP-9 in subcutaneous soft tissue， and the contents of transforming growth factor-β （TGF-β） and basic fibroblast growth factor （bFGF） in serum were investigated. RESULTS Chonghe paste had varying degrees of bacteriostatic effect on the above 4 bacteria （except for S. pneumoniae）， especially on S. aureus. Compared with the model group， on the 7th day of treatment， collagen fibers in the Chonghe paste group were arranged in an orderly manner， pus dissipated faster； the protein expressions of type Ⅰ and type Ⅲ collagen and the contents of TGF-β and bFGF were up-regulated significantly， while protein expressions of MMP-2 and MMP-9 were decreased significantly （P＜0.05）. On the 14th day of administration， collagen deposition was obvious in the Chonghe paste group， subcutaneous appendages gradually formed； the protein expressions of type Ⅰ and type Ⅲ collagen and the contents of TGF-β and bFGF were down-regulated significantly， while the protein expressions of MMP-2 and MMP-9 were increased significantly （P＜0.05）. CONCLUSIONS Chonghe paste has the bacteriostatic effect and may play a role in promoting the dissipation of swollen lesions by regulating the formation and decomposition of fibrin and increasing the secretion of bFGF and TGF-β.

Advances of Engineered Hydrogel Organoids within the Stem Cell Field: A Systematic Review.

Organoids are novel in vitro cell culture models that enable stem cells (including pluripotent stem cells and adult stem cells) to grow and undergo self-organization within a three-dimensional microenvironment during the process of differentiation into target tissues. Such miniature structures not only recapitulate the histological and genetic characteristics of organs in vivo, but also form tissues with the capacity for self-renewal and further differentiation. Recent advances in biomaterial technology, particularly hydrogels, have provided opportunities to improve organoid cultures; by closely integrating the mechanical and chemical properties of the extracellular matrix microenvironment, with novel synthetic materials and stem cell biology. This systematic review critically examines recent advances in various strategies and techniques utilized for stem-cell-derived organoid culture, with particular emphasis on the application potential of hydrogel technology in organoid culture. We hope this will give a better understanding of organoid cultures for modelling diseases and tissue engineering applications.

Implementation of an interactive virtual microscope laboratory system in teaching oral histopathology.

Laboratory course acts as a key component of histopathology education. Recent trends of incorporating visual and interactive technology in active and inquiry-based learning pedagogical methods have led to significant improvement of histopathology laboratory courses. The present work aimed to describe interactive virtual microscope laboratory system (IVMLS) as a virtual platform for teaching histopathology in order to improve the quality and efficiency of teaching. The system is based on interactive technology and consists of interactive software, slide-reading software, teaching resources and integrated auxiliary equipment. It allows real-time interaction between teachers and students and provides students with a wealth of learning and review materials. In order to evaluate the effectiveness of the system, we conducted a comparative study with the use of light microscope (LM) as a method. Specifically, we compared the results of six assignments and one laboratory final exam between IVMLS group and LM group to analyse the impact of IVMLS on students' academic performance. A questionnaire survey was also conducted to obtain students' attitudes and views on this system. There was no overall difference in assignment performance between IVMLS group and LM group. But laboratory final test grades increased from a mean of 62% (43.8-80.0, 95% CI) before to 83% (71.0-94.2, 95% CI) after implement IVMLS, suggesting highly significant (p < 0.001) improvement on students' histopathology laboratory performance. Feedback of the questionnaire was positive, indicating that students were satisfied with the system, which they believed improved student communication and teacher-student interaction, increased learning resources, increased their focus on learning, and facilitated their independent thinking process. This study proves that IVMLS is an efficient and feasible teaching technology and improves students' academic performance.

Effects of Postmortem Hemolysis and Ultrafiltration on Creatinine Detection Results / 法医学杂志

OBJECTIVES@#To investigate the interference of postmortem hemolysis on the detection of creatinine and whether ultrafiltration can reduce the interference.@*METHODS@#A total of 33 non-hemolyzed whole blood samples from the left heart were collected. Hemolyzed samples with 4 hemoglobin mass concentration gradients H1-H4 were artificially prepared. Ultrafiltration was performed on each hemolyzed sample. Creatinine concentrations in non-hemolyzed serum (baseline serum), hemolyzed samples and ultrafiltrate were detected. Bias (B), Pearson correlation and receiver operator characteristic (ROC) of baseline creatinine concentration between before and after ultrafiltration were analyzed.@*RESULTS@#As the hemoglobin mass concentration increased, B of the hemolyzed samples in the H1-H4 groups gradually increased, the |B| was 2.41(0.82, 8.25)-51.31(41.79, 188.25), reaching a maximum of 589.06%, and there was no statistically significant between the creatinine concentration and the baseline creatinine concentration (P=0.472 7, r=0.129 5). After ultrafiltration of hemolyzed samples, the interference of creatinine concentration in ultrafiltrate was significantly reduced, the |B| was 5.32(2.26, 9.22)-21.74(20.06, 25.58), reaching a maximum of 32.14%, and there was a positive correlation with baseline creatinine concentration (P<0.05, r=0.918 2). In the hemolyzed samples of H3 and H4 groups, there were 7 false-positive samples and 1 false-negative sample; in the ultrafiltrate samples, there were no false-positive sample and 1 false-negative sample. ROC analysis results showed the hemolyzed samples were lack of diagnostic value (P=0.117 5).@*CONCLUSIONS@#The postmortem hemolysis significantly interferes creatinine detection results of blood samples, ultrafiltration can reduce hemolysis-induced interference in postmortem creatinine detection.

Stability of IgE in Postmortem Plasma and Hemolyzed Samples / 法医学杂志

Objective To investigate the stability of IgE in postmortem plasma and hemolyzed samples under different storage conditions and freezing-thawing. Methods Thirty nine cardiac blood samples were collected from non-frozen corpses with the postmortem interval of less than 48 hours, including 20 plasma samples and 19 hemolyzed samples taken from whole blood. The samples were stored at -20 ℃, 4 ℃ and 25 ℃ for 28 d and at -80 ℃ for 1 year to evaluate the stability of IgE under different storage conditions. Repeated freezing-thawing treatment was conducted for 5 times to explore the stability of IgE in postmortem plasma and hemolyzed samples. IgE concentration in plasma and hemolyzed samples was detected by electroluminescence before and after treatment. Results The degradation rates of IgE in plasma samples under the three storage conditions, -20 ℃, 4 ℃ and 25 ℃ were close. After 28 d, the mean value was about 15%, the degradation speed of IgE in hemolyzed samples was faster than that of plasma under the same condition （P<0.05） and the degradation rate was faster than other two conditions under 25 ℃ （P<0.05）. The differences in the concentration of plasma samples after freezing at -80 ℃ for 1 year and that before freezing had no statistical significance （ P>0.05）, while the concentration of hemolyzed samples was degraded after freezing at -80 ℃ for 1 year （P<0.05）. The differences between the detection results of plasma and hemolyzed samples after repeated freezing-thawing for 5 times and that before freezing-thawing showed no statistical significance （ P>0.05）. Conclusion IgE has good freezing-thawing stability in postmortem plasma and hemolyzed samples. Stability of IgE is better in postmortem plasma samples than hemolyzed samples, thus it is recommended to separate plasma from postmortem blood samples as soon as possible in forensic practice.

Efficacy of Spleen-invigorating Chinese Medicinals in Repairing Intestinal Mucosal Barrier Injury： A Review / 中国实验方剂学杂志

As the first line of defense between the intestinal environment and the outside world， the intestinal mucosal barrier is essential for maintaining the intestinal homeostasis. The intestinal mucosal barrier injury will change the intestinal permeability and allow bacterial translocation and the entry of endotoxins into blood， thus triggering a series of inflammatory responses， followed by the injury of related tissues and the aggravation of primary diseases. The spleen， the acquired foundation， is responsible for maintaining the internal and external balance of the body and resisting external evils. Its physiological function is similar to that of the intestinal mucosal barrier. Spleen deficiency easily leads to intestinal mucosal barrier dysfunction. Therefore， replenishing Qi， invigorating spleen， and restoring the efficacy of spleen and stomach qi in defensing and governing transportation and transformation are the keys to prevent and treat intestinal mucosal barrier injury. In recent years， studies have shown that the spleen-invigorating Chinese medicinals repair the intestinal mucosal injury by promoting the expression of intestinal epithelial tight junction proteins， regulating the intestinal immune function， microbial flora， and metabolites， and supplementing the intestinal nutrition， enabling them to gradually become a research hotspot. After reviewing the relevant articles published in China and abroad， this paper expounded the common syndrome types of traditional Chinese medicine （TCM）， the changes in intestinal mucosal barrier induced by spleen deficiency， the repairing effects of spleen-invigorating Chinese medicinals on intestinal mucosal barrier injury， in order to provide some clues for the research on the treatment of intestinal mucosal barrier dysfunction-related diseases with spleen-invigorating Chinese medicinals.

Osteosynthesis of segmental alveolar fractures by internal fixation and the prognosis of the fractures and teeth.

BACKGROUND/AIMS: Fracture of the alveolar process is a common injury, but the traditional splinting fixation may not be possible in some cases. The aim of this study was to describe the osteosynthesis performed in segmental alveolar fractures by internal fixation and evaluate the prognosis of the fractures and teeth involved. MATERIALS AND METHODS: Twenty-two patients who were identified as having segmental alveolar fractures treated with osteosynthesis by internal fixation at the authors' department from January 2007 to December 2016 were included with 90 traumatized teeth. The prognosis of the fractures and teeth involved in the fractures was evaluated by the post-operative computed tomography combined with a follow-up study. RESULTS: All patients achieved healing and consolidation of the alveolar fractures. Furthermore, the occlusion was restored and the wounds healed. During the surgical procedures, no observed iatrogenic dental damage caused by the interdental drilling was found. Eventually, only 15.6% of the teeth had pulp necrosis, whereas the other healing complications were rare or not observed in the study. CONCLUSIONS: Osteosynthesis by internal fixation is an effective and safe treatment for some segmental alveolar fractures. The teeth involved in these fractures also have good prognosis.

Tibetan medicine Ru-yi-Zhen-bao Pills exhibits anti-migraine effect through mediating PAG anti-nociceptive channel.

ETHNOPHARMACOLOGICAL RELEVANCE: Migraine is a disabling neurovascular disorder, which increases risk of cardiovascular events and is a social burden worldwide. The present first-line anti-migraine medications can cause overwhelming side-effects, of which one includes the onset of cardiovascular disease. As one of the marketed Tibetan drugs, Ru-yi-Zhen-bao Pills (RYZBP) have been clinically used to treat cardiovascular disorders and as anti-migraine medication. However, there is currently no research exploring the anti-migraine actions of RYZBP. AIM OF THE STUDY: The current research was designed to assess the anti-migraine roles of RYZBP and explore the underlying mechanisms in a nitroglycerin (NTG)-induced migraine rat model trial. MATERIALS AND METHODS: 120 rats were randomly divided into the following six groups of 20 rats each: normal control group, model control group, positive control group, and RYZBP high/medium/low-dose groups (Ru-yi-Zhen-bao Pills; TH 1.00 g/kg, TM 0.50 g/kg and TL 0.25 g/kg). All rats were administered intragastrically for 7 consecutive days, which were subcutaneously injected with the NTG (10 mg/kg) after the last gavage (except in the normal control group). 3min after NTG treatment, 30 rats (5 rats from each group) were anesthetized and devoted to electroencephalogram(EEG) testing, which was used to evaluate the analgesic effect of RYZBP. One hour after NTG treatment, the rest of the 90 rats (15 rats from each group) were anesthetized and midbrain tissue sample was dissected. The dissection was then washed with physiological saline and collected. The histopathological changes in the periaqueductal gray(PAG) of 5 tissue samples were determined by aematoxylin-eosin (H&E) staining, as well as an estimation of substance P (SP) and neurokinin 1 receptor (NK1R) expression through immunohistochemically staining(IHC). Another 5 midbrain preparations were carried out to evaluate calcitonin gene-related peptide (CGRP), proenkephalin (PENK), SP, and cholecystokinin (CCK) expressions by real-time quantitative polymerase chain reaction (RT-qPCR). The rest of the 5 brainstem tissues were then used to measure CCK, CGRP, and opioid peptide receptor (DORR) levels by western blotting(WB). RESULTS: In the EEG test, RYZBP (TM 0.50 g / kg) treatment transformed the EEG pain-wave of the NTG-induced migraine model rats in different time period. In the mechanism assay, compared with the model control group, RYZBP pretreatment reduced inflammatory cell infiltration, fibrosis and vacuolation of neuronal cells of PAG tissue seen by HE staining. IHC experiments further showed that RYZBPTM up-regulated SP expression levels and enhanced NK1R levels in the NTG-induced migraine rats (P < 0.05). Therapeutic administration of RYZBP also increased PENK mRNA expression and DORR protein level. Both RT-qPCR and western blotting trials indicated that RYZBP treatment significantly decreased CCK and CGRP expression levels (P < 0.01 or P < 0.05) in the NTG-induced migraine rats. CONCLUSIONS: RYZBP has the potential to be an effective anti-migraine treatment through suppressing the EEG pain-wave, increasing the levels of SP, PENK, DORR and reducing expression of CCK and CGRP. Mediating the PAG anti-nociceptive channel and inhibiting central sensitization were the two potential mechanisms, which offers further evidence for clinical therapy.

Application of Hollow Fiber Ultrafiltration Technology in the Detection of Total IgE of Hemolytic Blood after Death / 法医学杂志

Objective To investigate the treatment effect of hollow fiber ultrafiltration technology on hemolytic samples and the differences between IgE concentration and serum concentration before hemolysis in ultrafiltrate. Methods The 33 postmortem blood samples of non-frozen corpses within 72 hours after death were collected, 4 mL blood was taken from each case, among which 1 mL was centrifuged to get serum, and the remaining 3 mL blood was frozen-thawed 3-5 times to cause complete hemolysis. The 2 mL hemolytic samples were processed by hollow fiber ultrafiltration to obtain ultrafiltrate. The hemoglobin concentration in serum, complete hemolytic sample and ultrafiltrate was determined by Van-Zij solution-cyanated methemoglobin assay method, and the total IgE in serum and ultrafiltrate was determined by electrochemical luminescence method. Results The hemoglobin concentration in ultrafiltrate was significantly lower than that in complete hemolytic samples （P<0.05）. There was a good correlation between the total IgE detection values of ultrafiltrate and serum （r=0.984）. The difference between the serum and the value of IgE in ultrafiltrate after correction had no statistical significance, and the differences between the two in positive rates had no statistical significance （P>0.05）. Conclusion Ultrafiltration technology has a good treatment effect on complete hemolytic samples, and the correction value of ultrafiltrate detection is close to the serum level before hemolysis, and therefore, it can be applied to the detection of total IgE of frozen corpse hemolytic samples.

MicroRNA-495 downregulates AQP1 and facilitates proliferation and differentiation of osteoblasts in mice with tibial fracture through activation of p38 MAPK signaling pathway.

Osteoblasts are implicated in the building of the vertebrate skeleton. The current study aimed to investigate the role of microRNA-495 (miR-495) in the osteoblasts of mice with tibial fractures and the underlying mechanism involving in aquaporin-1 (AQP1) and the p38 mitogen-activated protein kinase (p38 MAPK) signaling pathway. Initially, a microarray-based analysis was performed to screen the differentially expressed genes and miRNAs associated with tibial fracture. Following the establishment of a tibial fracture mouse model, the positive rate of the AQP1 protein in the fracture tissue was detected by immunohistochemistry (IHC). Next, to verify the binding site between miR-495 on AQP1, bioinformatics data were employed in addition to the application of a dual-luciferase reporter gene assay. The osteoblast cell line MC3T3-E1 was treated with miR-495 mimic, miR-495 inhibitor and Anisomycin to explore the potent effects of miR-495 on proliferation and differentiation of osteoblasts in mice with tibial fracture. The expression of miR-495, AQP1, p38 MAPK, PCNA, Cyclin D1, OCN, and OPN was subsequently evaluated by RT-qPCR and Western blot analysis. Cell viability, the number of calcium nodules and alkaline phosphatase (ALP) activity were detected by MTT assay, alizarin red staining, and ALP activity assay, respectively. Our results revealed that miR-495 was down-regulated while AQP1 was up-regulated in the mice with tibial fractures. AQP1 was verified as a target gene of miR-495. When the cells were treated with overexpressed miR-495 or activated p38 MAPK signaling pathway, elevated expression of PCNA, Cyclin, D1, OCN, and OPN along with an increased amount of calcium nodules, higher cell viability, and enhanced ALP activity was detected, while the expression of AQP1 was reduced. Collectively, the key findings of the present study support the notion that overexpressed miR-495 may activate the p38 MAPK signaling pathway to inhibit AQP1 and to promote the proliferation and differentiation of osteoblasts in mice with tibial fracture.

Effect of Corpse Cryopreservation on Forensic Pathological Identification / 法医学杂志

Because cryopreservation can effectively slow down the putrefaction and prolong the preservation time of corpses, it has become the main way of corpse preservation in China. However, it may cause a certain degree of non-specific effects on the corpses and thus interfere with forensic pathological identification. This paper summarizes relevant problems reported in domestic and foreign literature reports and practical identification, and analyzes the effects of cryopreservation on corpses from aspects of anatomical findings, histomorphology, postmortem biochemistry, and postmortem imaging, therefore to provide Chinese forensic workers assistance on problems in their practice of forensic pathology.