Reduced miR-99a-3p levels in systemic lupus erythematosus may promote B cell proliferation via NCAPG and the PI3K/AKT signaling pathway.

BACKGROUND: Systemic lupus erythematosus is an immunologically dysregulated disease characterized by the presence of multiple autoantibodies. In SLE, B lymphocytes contribute to the dysregulated production of autoantibodies and cytokines. Recently, we discovered that miR-99a-3p binds to both EIF4EBP1 and NCAPG mRNA and that lowering miR-99a-3p can promote B cell autophagy in SLE by increasing EIF4EBP1 expression. However, the functions of miR-99a-3p and NCAPG in SLE have not been extensively investigated. OBJECTIVE: This work aims to evaluate the levels of miR-99a-3p and NCAPG expression in SLE B cells and to determine whether the aberrant expression of miR-99a-3p and NCAPG contributes to the pathological mechanisms in SLE. METHODS: B lymphocytes were obtained through immunomagnetic negative selection. Using RT-qPCR, miR-99a-3p and NCAPG mRNA expressions in B lymphocytes and in the BALL-1 cell line were measured. To determine the relative abundance of NCAPG, PI3K, p-PI3K, AKT, and p-AKT, we normalize them to the level of ß-actin using Western blotting. Evaluation of miR-99a-3p and NCAPG's impact on cell proliferation was done utilizing CCK-8 assay. Using flow cytometry, the cell cycle and apoptosis were both measured. RESULTS: Comparing SLE B cells to healthy controls, miR-99a-3p expression was significantly downregulated. Additionally, it was observed that SLE B cells had significantly higher NCAPG mRNA expression. Blocking miR-99a-3p expression in BALL-1 cells with an antagomir elevated NCAPG expression, facilitated PI3K/AKT pathway activation, improved cell proliferation, raised the fraction of S-phase cells, and prevented cell apoptosis. The opposite effects of upregulated miR-99a-3p levels on BALL-1 cells were observed by using an agomir. Furthermore, the effect of decreased miR-99a-3p expression on cell proliferation was partially mediated by elevating NCAPG levels and activating the PI3K/AKT pathway. CONCLUSION: Our research indicates that lower miR-99a-3p expression in SLE B cells appears to boost B cell number via the NCAPG and PI3K/AKT pathways.

Estimating the Number of Communities in Weighted Networks.

Community detection in weighted networks has been a popular topic in recent years. However, while there exist several flexible methods for estimating communities in weighted networks, these methods usually assume that the number of communities is known. It is usually unclear how to determine the exact number of communities one should use. Here, to estimate the number of communities for weighted networks generated from arbitrary distribution under the degree-corrected distribution-free model, we propose one approach that combines weighted modularity with spectral clustering. This approach allows a weighted network to have negative edge weights and it also works for signed networks. We compare the proposed method to several existing methods and show that our method is more accurate for estimating the number of communities both numerically and empirically.

M2 macrophages mediate fibrotic scar formation in the early stages after cerebral ischemia in rats.

In the central nervous system, the formation of fibrotic scar after injury inhibits axon regeneration and promotes repair. However, the mechanism underlying fibrotic scar formation and regulation remains poorly understood. M2 macrophages regulate fibrotic scar formation after injury to the heart, lung, kidney, and central nervous system. However, it remains to be clarified whether and how M2 macrophages regulate fibrotic scar formation after cerebral ischemia injury. In this study, we found that, in a rat model of cerebral ischemia induced by middle cerebral artery occlusion/reperfusion, fibrosis and macrophage infiltration were apparent in the ischemic core in the early stage of injury (within 14 days of injury). The number of infiltrated macrophages was positively correlated with fibronectin expression. Depletion of circulating monocyte-derived macrophages attenuated fibrotic scar formation. Interleukin 4 (IL4) expression was strongly enhanced in the ischemic cerebral tissues, and IL4-induced M2 macrophage polarization promoted fibrotic scar formation in the ischemic core. In addition, macrophage-conditioned medium directly promoted fibroblast proliferation and the production of extracellular matrix proteins in vitro. Further pharmacological and genetic analyses showed that sonic hedgehog secreted by M2 macrophages promoted fibrogenesis in vitro and in vivo, and that this process was mediated by secretion of the key fibrosis-associated regulatory proteins transforming growth factor beta 1 and matrix metalloproteinase 9. Furthermore, IL4-afforded functional restoration on angiogenesis, cell apoptosis, and infarct volume in the ischemic core of cerebral ischemia rats were markedly impaired by treatment with an sonic hedgehog signaling inhibitor, paralleling the extent of fibrosis. Taken together, our findings show that IL4/sonic hedgehog/transforming growth factor beta 1 signaling targeting macrophages regulates the formation of fibrotic scar and is a potential therapeutic target for ischemic stroke.

Estimating Mixed Memberships in Directed Networks by Spectral Clustering.

Community detection is an important and powerful way to understand the latent structure of complex networks in social network analysis. This paper considers the problem of estimating community memberships of nodes in a directed network, where a node may belong to multiple communities. For such a directed network, existing models either assume that each node belongs solely to one community or ignore variation in node degree. Here, a directed degree corrected mixed membership (DiDCMM) model is proposed by considering degree heterogeneity. An efficient spectral clustering algorithm with a theoretical guarantee of consistent estimation is designed to fit DiDCMM. We apply our algorithm to a small scale of computer-generated directed networks and several real-world directed networks.

Asymmetric activation of microglia in the hippocampus drives anxiodepressive consequences of trigeminal neuralgia in rodents.

BACKGROUND AND PURPOSE: Patients suffering from trigeminal neuralgia are often accompanied by anxiety and depression. Microglia-mediated neuroinflammation is involved in the development of neuropathic pain and anxiodepression pathogenesis. Whether and how microglia are involved in trigeminal neuralgia-induced anxiodepression remains unclear. EXPERIMENTAL APPROACH: Unilateral constriction of the infraorbital nerve (CION) was performed to establish trigeminal neuralgia in rat and mouse models. Mechanical allodynia and anxiodepressive-like behaviours were measured. Optogenetic and pharmacological manipulations were employed to investigate the role of hippocampal microglia in anxiety and depression caused by trigeminal neuralgia. KEY RESULTS: Trigeminal neuralgia activated ipsilateral but not contralateral hippocampal microglia, up-regulated ipsilateral hippocampal ATP and interleukin-1ß (IL-1ß) levels, impaired ipsilateral hippocampal long-term potentiation (LTP) and induced anxiodepressive-like behaviours in a time-dependent manner in rodents. Pharmacological or optogenetic inhibition of ipsilateral hippocampal microglia completely blocked trigeminal neuralgia-induced anxiodepressive-like behaviours. Activation of unilateral hippocampal microglia directly elicited an anxiodepressive state and impaired hippocampal LTP. Knockdown of ipsilateral hippocampal P2X7 receptors prevented trigeminal neuralgia-induced microglial activation and anxiodepressive-like behaviours. Furthermore, we demonstrated that microglia-derived IL-1ß mediated microglial activation-induced anxiodepressive-like behaviours and LTP impairment. CONCLUSION AND IMPLICATIONS: These findings suggest that priming of microglia with ATP/P2X7 receptors in the ipsilateral hippocampus drives pain-related anxiodepressive-like behaviours via IL-1ß. An asymmetric role of the bilateral hippocampus in trigeminal neuralgia-induced anxiety and depression was uncovered. The approaches targeting microglia and P2X7 signalling might offer novel therapies for trigeminal neuralgia-related anxiety and depressive disorder.

TMEM16F may be a new therapeutic target for Alzheimer's disease.

TMEM16F is involved in many physiological processes such as blood coagulation, cell membrane fusion and bone mineralization. Activation of TMEM16F has been studied in various central nervous system diseases. High TMEM16F level has been also found to participate in microglial phagocytosis and transformation. Microglia-mediated neuroinflammation is a key factor in promoting the progression of Alzheimer's disease. However, few studies have examined the effects of TMEM16F on neuroinflammation in Alzheimer's disease. In this study, we established TMEM16F-knockdown AD model in vitro and in vivo to investigate the underlying regulatory mechanism about TMEM16F-mediated neuroinflammation in AD. We performed a Morris water maze test to evaluate the spatial memory ability of animals and detected markers for the microglia M1/M2 phenotype and NLRP3 inflammasome. Our results showed that TMEM16F was elevated in 9-month-old APP/PS1 mice. After TMEM16F knockdown in mice, spatial memory ability was improved, microglia polarization to the M2 phenotype was promoted, NLRP3 inflammasome activation was inhibited, cell apoptosis and Aß plaque deposition in brain tissue were reduced, and brain injury was alleviated. We used amyloid-beta (Aß25-35) to stimulate human microglia to construct microglia models of Alzheimer's disease. The levels of TMEM16F, inducible nitric oxide synthase (iNOS), proinflammatory cytokines and NLRP3 inflammasome-associated biomarkers were higher in Aß25-35 treated group compared with that in the control group. TMEM16F knockdown enhanced the expression of the M2 phenotype biomarkers Arg1 and Socs3, reduced the release of proinflammatory factors interleukin-1, interleukin-6 and tumor necrosis factor-α, and inhibited NLRP3 inflammasome activation through reducing downstream proinflammatory factors interleukin-1ß and interleukin-18. This inhibitory effect of TMEM16F knockdown on M1 microglia was partially reversed by the NLRP3 agonist Nigericin. Our findings suggest that TMEM16F participates in neuroinflammation in Alzheimer's disease through participating in polarization of microglia and activation of the NLRP3 inflammasome. These results indicate that TMEM16F inhibition may be a potential therapeutic approach for Alzheimer's disease treatment.

Degree-corrected distribution-free model for community detection in weighted networks.

A degree-corrected distribution-free model is proposed for weighted social networks with latent structural information. The model extends the previous distribution-free models by considering variation in node degree to fit real-world weighted networks, and it also extends the classical degree-corrected stochastic block model from un-weighted network to weighted network. We design an algorithm based on the idea of spectral clustering to fit the model. Theoretical framework on consistent estimation for the algorithm is developed under the model. Theoretical results when edge weights are generated from different distributions are analyzed. We also propose a general modularity as an extension of Newman's modularity from un-weighted network to weighted network. Using experiments with simulated and real-world networks, we show that our method significantly outperforms the uncorrected one, and the general modularity is effective.

Studying Asymmetric Structure in Directed Networks by Overlapping and Non-Overlapping Models.

We consider the problem of modeling and estimating communities in directed networks. Models to this problem in the previous literature always assume that the sending clusters and the receiving clusters have non-overlapping property or overlapping property simultaneously. However, previous models cannot model the directed network in which nodes in sending clusters have overlapping property, while nodes in receiving clusters have non-overlapping property, especially for the case when the number of sending clusters is no larger than that of the receiving clusters. This kind of directed network exists in the real world for its randomness, and by the fact that we have little prior knowledge of the community structure for some real-world directed networks. To study the asymmetric structure for such directed networks, we propose a flexible and identifiable Overlapping and Non-overlapping model (ONM). We also provide one model as an extension of ONM to model the directed network, with a variation in node degree. Two spectral clustering algorithms are designed to fit the models. We establish a theoretical guarantee on the estimation consistency for the algorithms under the proposed models. A small scale computer-generated directed networks are designed and conducted to support our theoretical results. Four real-world directed networks are used to illustrate the algorithms, and the results reveal the existence of highly mixed nodes and the asymmetric structure for these networks.

A Useful Criterion on Studying Consistent Estimation in Community Detection.

In network analysis, developing a unified theoretical framework that can compare methods under different models is an interesting problem. This paper proposes a partial solution to this problem. We summarize the idea of using a separation condition for a standard network and sharp threshold of the Erdös-Rényi random graph to study consistent estimation, and compare theoretical error rates and requirements on the network sparsity of spectral methods under models that can degenerate to a stochastic block model as a four-step criterion SCSTC. Using SCSTC, we find some inconsistent phenomena on separation condition and sharp threshold in community detection. In particular, we find that the original theoretical results of the SPACL algorithm introduced to estimate network memberships under the mixed membership stochastic blockmodel are sub-optimal. To find the formation mechanism of inconsistencies, we re-establish the theoretical convergence rate of this algorithm by applying recent techniques on row-wise eigenvector deviation. The results are further extended to the degree-corrected mixed membership model. By comparison, our results enjoy smaller error rates, lesser dependence on the number of communities, weaker requirements on network sparsity, and so forth. The separation condition and sharp threshold obtained from our theoretical results match the classical results, so the usefulness of this criterion on studying consistent estimation is guaranteed. Numerical results for computer-generated networks support our finding that spectral methods considered in this paper achieve the threshold of separation condition.

Evidence for a mouse origin of the SARS-CoV-2 Omicron variant

The rapid accumulation of mutations in the SARS-CoV-2 Omicron variant that enabled its outbreak raises questions as to whether its proximal origin occurred in humans or another mammalian host. Here, we identified 45 point mutations that Omicron acquired since divergence from the B.1.1 lineage. We found that the Omicron spike protein sequence was subjected to stronger positive selection than that of any reported SARS-CoV-2 variants known to evolve persistently in human hosts, suggesting the possibility of host-jumping. The molecular spectrum (i.e., the relative frequency of the twelve types of base substitutions) of mutations acquired by the progenitor of Omicron was significantly different from the spectrum for viruses that evolved in human patients, but was highly consistent with spectra associated with evolution in a mouse cellular environment. Furthermore, mutations in the Omicron spike protein significantly overlapped with SARS-CoV-2 mutations known to promote adaptation to mouse hosts, particularly through enhanced spike protein binding affinity for the mouse cell entry receptor. Collectively, our results suggest that the progenitor of Omicron jumped from humans to mice, rapidly accumulated mutations conducive to infecting that host, then jumped back into humans, indicating an inter-species evolutionary trajectory for the Omicron outbreak.

Nanopore Formation and Structural Changes in Black Shale During the Initial Weathering Stage: A Longmaxi Formation Profile in Northwestern Hunan, China.

Understanding the controls on composition changes and porosity evolution in the critical zone of shale remains a major challenge. The aim of the present study is to develop a model of the changes in mineral compositions, chemical compositions and nanopore formation in shale during the initial weathering stage. To understand these processes, we selected a Silurian shale profile rich in pyrite and organic matter located in South China. Based on X-ray diffraction (XRD) and bulk elemental data, the variations in mineralogical and chemical compositions with depth were studied. To characterize the full pore size spectrum and to gain insight into the nature of secondary pores and their relationship with weathering, nuclear magnetic resonance (NMR) measurements and petrographic observations were combined with scanning electron microscopy (SEM) imaging. The results show that Al and K are enriched slightly, while Ca and Na are depleted in the upper part of the weathering profile. Si, Mn and Ti are relatively stable from the bottom to the top of the profile. Quartz, feldspar, mica, illite and chlorite are the main minerals in the parent rock, and they are relatively stable along the profile. The rock density gradually decreases from 2.6 g/cm³ to 2.1 g/cm³ from the bottom to the top, and the color of the shales changes from black to grayish yellow, but no secondary minerals are detected. The chemical weathering of black shale is dominated by the oxidation of pyrite and organic matter, giving rise to color variation and nanopore formation. The increase in interparticle pores at the nanometer-micron scale is initiated by the dissolution of easily weathered components such as organic matter and pyrite. The removal of clay minerals and tiny particles by groundwater seepage may be the main cause of porosity enhancement during the initial weathering stage. This study suggests that nanoporosity may play an important role in the process of fluid-rock interaction within black shale during the initial weathering stage.

A fixed nitrous oxide/oxygen mixture as an analgesic for trauma patients in emergency department: study protocol for a randomized, controlled trial.

BACKGROUND: Acute pain is always the most common complaint in Emergency Department admissions and options for analgesia are limited. Nitrous oxide/oxygen possess many properties showing it may be an ideal analgesic method for the Emergency Department; it is quick-acting, well-tolerated, and does not mask signs and symptoms. The aim of this study is to evaluate the safety and analgesic effect of the fixed nitrous oxide/oxygen mixture for trauma patients in a busy emergency environment. METHODS: The randomized, double-blind, prospective, placebo-controlled study will be carried out in the Emergency Department of General Hospital of Ningxia Medical University. The target research objects are trauma patients who present to the Emergency Department and report moderate to severe intensities of acute pain. A total of 90 patients will be recruited and randomly assigned into the treatment and control group. The treatment group will receive conventional pain treatment plus nitrous oxide/oxygen mixture and the control group will receive conventional pain treatment plus oxygen. Neither patients, nor investigators, nor data collectors will know the nature of the gas mixture in each cylinder and the randomization list. Outcomes will be monitored at baseline(T0), 5 min (T1), and 15 min (T2) after the beginning of intervention and at 5 min post intervention (T3) for each group. The primary outcome is the level of pain relief after the initial administering of the intervention at T1, T2, and T3. Secondary outcomes include adverse events, physiological parameters, total time of the gas administration, satisfaction from both patients and healthcare professionals, and the acceptance of patients. DISCUSSION: Our previous studies suggested that a fixed nitrous oxide/oxygen mixture was an efficacious analgesic for the management of burning dressing pain and breakthrough cancer pain. The results of this study will provide a more in-depth understanding of the effect of this gas. If this treatment proves successful, it could help to generate preliminary guidelines and be implemented widely in trauma patients with pain in Emergency Departments. TRIAL REGISTRATION: Chinese Clinical Trial Register, ChiCTR-INR-16007807 . Registered on 21 January 2016.

Bidirectional modulation between infiltrating CD3+ T-lymphocytes and astrocytes in the spinal cord drives the development of allodynia in monoarthritic rats.

Increasing evidence suggests that T cells and glia participate in the process of neuropathic pain. However, little is known about the involvement of T cells or the interaction between glia and T cells at the molecular level. Here we investigated the phenotype of T cell infiltration into the spinal cord in inflammatory pain and explored potential crosstalk between glia and T cells. The establishment of monoarthritis produced T cell infiltration and astrocyte activation, exhibiting similar kinetics in the spinal cord. T-cell-deficient (Rag1-/-) mice significantly attenuated MA-induced mechanical allodynia and GFAP upregulation. Double immunofluorescence staining showed that CD3 mainly colocalized with interferon-gamma (IFN-γ). Western blot and flow cytometry showed that multiple intrathecal administrations of astrocytic inhibitor fluorocitrate decreased IFN-γ-production without decreasing T cell number in the spinal cord. Spinal IFN-γ blockade reduced MA-induced mechanical allodynia and astroglial activation. In contrast, treatment with rIFN-γ directly elicited persistent mechanical allodynia and upregulation of GFAP and pJNK1/2 in naïve rats. Furthermore, rIFN-γ upregulated the phosphorylation of NF-κB p65 in cultured astrocytes vitro and spinal dorsal horn vivo. The results suggest that Th1 cells and astrocytes maintain inflammatory pain and imply that there may be a positive feedback loop between these cells via IFN-γ.

Proteomic Analysis of the Hippocampus in Mouse Models of Trigeminal Neuralgia and Inescapable Shock-Induced Depression.

To investigate the behavioral and biomolecular similarity between neuralgia and depression, a trigeminal neuralgia (TN) mouse model was established by constriction of the infraorbital nerve (CION) to mimic clinical trigeminal neuropathic pain. A mouse learned helplessness (LH) model was developed to investigate inescapable foot-shock-induced psychiatric disorders like depression in humans. Mass spectrometry was used to assess changes in the biomolecules and signaling pathways in the hippocampus from TN or LH mice. TN mice developed not only significant mechanical allodynia but also depressive-like behaviors (mainly behavioral despair) at 2 weeks after CION, similar to LH mice. MS analysis demonstrated common and distinctive protein changes in the hippocampus between groups. Many protein function families (such as cell-to-cell signaling and interaction, and cell assembly and organization,) and signaling pathways (e.g., the Huntington's disease pathway) were involved in chronic neuralgia and depression. Together, these results demonstrated that the LH and TN models both develop depressive-like behaviors, and revealed the involvement of many psychiatric disorder-related biomolecules/pathways in the pathogenesis of TN and LH.

HeteroMeth: A Database of Cell-to-cell Heterogeneity in DNA Methylation / 基因组蛋白质组与生物信息学报·英文版

DNA methylation is an important epigenetic mark that plays a vital role in gene expression and cell differentiation. The average DNA methylation level among a group of cells has been extensively documented. However, the cell-to-cell heterogeneity in DNA methylation, which reflects the differentiation of epigenetic status among cells, remains less investigated. Here we established a gold standard of the cell-to-cell heterogeneity in DNA methylation based on single-cell bisulfite sequencing (BS-seq) data. With that, we optimized a computational pipeline for estimating the heterogeneity in DNA methylation from bulk BS-seq data. We further built HeteroMeth, a database for searching, browsing, visualizing, and downloading the data for heterogeneity in DNA methylation for a total of 141 samples in humans, mice, Arabidopsis, and rice. Three genes are used as examples to illustrate the power of HeteroMeth in the identification of unique features in DNA methylation. The optimization of the computational strategy and the construction of the database in this study complement the recent experimental attempts on single-cell DNA methylomes and will facilitate the understanding of epigenetic mechanisms underlying cell differentiation and embryonic development. HeteroMeth is publicly available at http://qianlab.genetics.ac.cn/HeteroMeth.

Proteomic Analysis of the Hippocampus in Mouse Models of Trigeminal Neuralgia and Inescapable Shock-Induced Depression / 神经科学通报·英文版

To investigate the behavioral and biomolecular similarity between neuralgia and depression, a trigeminal neuralgia (TN) mouse model was established by constriction of the infraorbital nerve (CION) to mimic clinical trigeminal neuropathic pain. A mouse learned helplessness (LH) model was developed to investigate inescapable foot-shock-induced psychiatric disorders like depression in humans. Mass spectrometry was used to assess changes in the biomolecules and signaling pathways in the hippocampus from TN or LH mice. TN mice developed not only significant mechanical allodynia but also depressive-like behaviors (mainly behavioral despair) at 2 weeks after CION, similar to LH mice. MS analysis demonstrated common and distinctive protein changes in the hippocampus between groups. Many protein function families (such as cell-to-cell signaling and interaction, and cell assembly and organization,) and signaling pathways (e.g., the Huntington's disease pathway) were involved in chronic neuralgia and depression. Together, these results demonstrated that the LH and TN models both develop depressive-like behaviors, and revealed the involvement of many psychiatric disorder-related biomolecules/pathways in the pathogenesis of TN and LH.

Contrast Researches of Liver Biopsy and Ultrasound in the Diagnosis of Fatty Liver / 昆明医科大学学报

Objective To evaluate the significance of liver biopsy and B ultrasonograpgy in the diagnosis of fatty liver. Methods The results of 62 patients with liver steatosis diagnosed by liver puncture biopsy but not by B-ultrasonograpgy were contrastively analyzed and combined with liver function, blood lipids, blood glucose, and body mass index. Results The 62 cases which were not diagnosed as fatty liver by B-ultrasonograpgy were proved to be 5％-33％liver steatosis after liver puncture biopsy. Among the 62 cases, 23 cases were indicated by the B-ultrasonograpgy that the liver parenchyma echo did not see abnormalities, 18 cases showed the liver parenchyma echo slightly was enlarged, 17 cases showed the liver parenchyma echo density was a bit enhanced and 4 cases were diffuse liver damage,which respectively were 37.01％、29.03％、27.42％and 6.45％. Pathologically it indicated that 45 cases were 5％≤liver steatosis≤19％. Among the 45 cases, 18 cases were indicated by the B-ultrasonograpgy that the liver parenchyma echo was not seen abnormalities, 8 cases showed the liver parenchyma echo slightly was enlarged, 17 cases showed the liver parenchyma echo density was a bit enhanced, and 2 cases were diffuse liver damage, and the change of ultrasound was mainly showed by the liver parenchyma echo not seen abnormalities and the enhanced liver parenchyma echo density. Pathologically it indicated that 17 cases were 20％≤liver steatosis≤33％, 6 cases were indicated by the B-ultrasonograpgy that the liver parenchyma echo did not see abnormalities, 5 cases showed the liver parenchyma echo slightly was enlarged, 5 cases showed the liver parenchyma echo density was a bit enhanced, and 1 case was diffuse liver damage, and the change of ultrasound was mainly showed by the liver parenchyma echo not seen abnormalities, the slightly enlarged liver parenchyma echo and the enhanced liver parenchyma echo density. By analyzing the influence to the ultrasound changes by patients' liver function, body mass index, blood fat and blood sugar, and with logistic regression analysis through a disorderly classification, it was found that the larger value of the glutamine transferase, alkaline phosphatase, body mass index, triglyceride and low density lipoprotein cholesterol, the higher possibility of diffuse liver damage, and the higher level of fatty degeneration, the higher possibility of diffuse liver damage. Conclusion In the diagnosis of fatty liver, when the fatty degeneration is below 1/3, B-ultrasonic examination can't show characteristic changes of fatty liver. It should be closely observed or take liver puncture biopsy to make a definite diagnosis of fatty liver.

ABT-263 induces G1/G0-phase arrest, apoptosis and autophagy in human esophageal cancer cells in vitro.

Both the anti- and pro-apoptotic members of the Bcl-2 family are regulated by a conserved Bcl-2 homology (BH3) domain. ABT-263 (Navitoclax), a novel BH3 mimetic and orally bioavailable Bcl-2 family inhibitor with high affinity for Bcl-xL, Bcl-2 and Bcl-w has entered clinical trials for cancer treatment. But the anticancer mechanisms of ABT-263 have not been fully elucidated. In this study we investigated the effects of ABT-263 on human esophageal cancer cells in vitro and to explore its anticancer mechanisms. Treatment with ABT-263 dose-dependently suppressed the viability of 3 human esophageal cancer cells with IC50 values of 10.7±1.4, 7.1±1.5 and 8.2±1.6 µmol/L, in EC109, HKESC-2 and CaES-17 cells, respectively. ABT-263 (5-20 µmol/L) dose-dependently induced G1/G0-phase arrest in the 3 cancer cell lines and induced apoptosis evidenced by increased the Annexin V-positive cell population and elevated levels of cleaved caspase 3, cleaved caspase 9 and PARP. We further demonstrated that ABT-263 treatment markedly increased the expression of p21Waf1/Cip1 and decreased the expression of cyclin D1 and phospho-Rb (retinoblastoma tumor suppressor protein) (Ser780) proteins that contributed to the G1/G0-phase arrest. Knockdown of p21Waf1/Cip1 attenuated ABT-263-induced G1/G0-phase arrest. Moreover, ABT-263 treatment enhanced pro-survival autophagy, shown as the increased LC3-II levels and decreased p62 levels, which counteracted its anticancer activity. Our results suggest that ABT-263 exerts cytostatic and cytotoxic effects on human esophageal cancer cells in vitro and enhances pro-survival autophagy, which counteracts its anticancer activity.

Analysis on the Characteristics of Insomnia Groups Based on Web Data Mining / 医学信息学杂志

The paper introduces some common methods (including web crawler technology,Chinese words segmentation and keyword extraction algorithm) of Web data mining,acquires the data related to insomnia in the online medical website through the web crawler technology,classifies and processes the data,carries out words segmentation and keywords extraction of the text data based on the rules,and analyzes the gender and age distribution situations,symptoms,causes of disease and other characteristics of patients with insomnia.

[Synergistic anti-tumor effect of obatoclax and MG-132 in esophageal cancer cell line CaES-17].

OBJECTIVE: To explore whether MG-132 could enhance the anti-tumor activity of obatoclax against esophageal cancer cell line CaES-17. METHODS: MTT assay was used to determine the cytotoxicity of obatoclax and MG-132 in CaES-17 cells. The IC(50) of obatoclax and MG-132 were used to determine the molar ratio (1:2.4) of the two drugs for combined treatment of the cells. The concentrations of obatoclax and MG-132 ranged from 1/8 IC(50) to 4 IC(50) after serial dilution, and their combination index (CI) was calculated using CompuSyn software. The expression of ubiquitin and the cleavage of PARP, caspase-9, phospho-histone H3 and phospho-aurora A/B/C in the exposed cells were examined with Western blotting; the cell apoptosis was measured by flow cytometry with Annexin V staining, and the percentage of cells in each cell cycle phase was also determined by flow cytometry. RESULTS: The CI of obatoclax and MG-132 was 0.296 for a 50% inhibition of Caes-17 cells and was 0.104 for a 95% inhibition. The cells treated with obatoclax or MG-132 alone showed increased expression of ubiquitin and cleavage of PARP and caspase-9. Compared with the cells treated with obatoclax or MG-132 alone, the cells with a combined treatment exhibited significantly increased expression of ubiquitin, cleavage of PARP and caspase-9, and expression of phospho-Histone H3 (P<0.05). The combined treatment of the cells also resulted in significantly increased expression of phospho-Aurora A/B/C compared with obatoclax treatment alone. The cells with the combined treatment showed significantly higher percentages of apoptotic cells and cells in sub-G(1) and G(2)/M phases compared with the cells treated with either of the drugs (P<0.05). CONCLUSION: Obatoclax combined with MG-132 shows a significant synergistic anti-tumor effect against esophageal cancer CaES-17 cells by inducing apoptosis and cell cycle arrest.