Radiogenomic analysis for predicting lymph node metastasis and molecular annotation of radiomic features in pancreatic cancer.

BACKGROUND: To provide a preoperative prediction model for lymph node metastasis in pancreatic cancer patients and provide molecular information of key radiomic features. METHODS: Two cohorts comprising 151 and 54 pancreatic cancer patients were included in the analysis. Radiomic features from the tumor region of interests were extracted by using PyRadiomics software. We used a framework that incorporated 10 machine learning algorithms and generated 77 combinations to construct radiomics-based models for lymph node metastasis prediction. Weighted gene coexpression network analysis (WGCNA) was subsequently performed to determine the relationships between gene expression levels and radiomic features. Molecular pathways enrichment analysis was performed to uncover the underlying molecular features. RESULTS: Patients in the in-house cohort (mean age, 61.3 years ± 9.6 [SD]; 91 men [60%]) were separated into training (n = 105, 70%) and validation (n = 46, 30%) cohorts. A total of 1,239 features were extracted and subjected to machine learning algorithms. The 77 radiomic models showed moderate performance for predicting lymph node metastasis, and the combination of the StepGBM and Enet algorithms had the best performance in the training (AUC = 0.84, 95% CI = 0.77-0.91) and validation (AUC = 0.85, 95% CI = 0.73-0.98) cohorts. We determined that 15 features were core variables for lymph node metastasis. Proliferation-related processes may respond to the main molecular alterations underlying these features. CONCLUSIONS: Machine learning-based radiomics could predict the status of lymph node metastasis in pancreatic cancer, which is associated with proliferation-related alterations.

Cortical tagged synaptic long-term depression in the anterior cingulate cortex of adult mice.

The anterior cingulate cortex (ACC) is a key cortical region for pain perception and emotion. Different forms of synaptic plasticity, including long-term potentiation (LTP) and long-term depression (LTD), have been reported in the ACC. Synaptic tagging of LTP plays an important role in hippocampus-related associative memory. In this study, we demonstrate that synaptic tagging of LTD is detected in the ACC of adult male and female mice. This form of tagged LTD requires the activation of metabotropic glutamate receptor subtype 1 (mGluR1). The induction of tagged LTD is time-related with the strongest tagged LTD appearing when the interval between two independent stimuli is 30 min. Inhibitors of mGluR1 blocked the induction of tagged LTD, however, blocking N-methyl-d-aspartate (NMDA) receptors did not affect the induction of tagged LTD. Nimodipine, an inhibitor of L-type voltage-gated calcium channels (VGCCs), also blocked tagged LTD. In an animal model of amputation, we found that tagged LTD was either reduced or completely blocked. Together with our previous report of tagged LTP in the ACC, this study strongly suggests that excitatory synapses in the adult ACC are highly plastic. The biphasic tagging of synaptic transmission provides a new form of heterosynaptic plasticity in the ACC which has functional and pathophysiological significance in phantom pain.Significance Statement The anterior cingulate cortex (ACC) is a key cortical region for pain perception and chronic pain. Previous studies have reported a novel form of long-term heterosynaptic potentiation in the ACC. In this study, we discovered a long-term depression (LTD) form of synaptic tagging in the ACC of adult male and female mice. This form of tagged LTD requires the activation of metabotropic glutamate receptors (mGluRs). In an animal model of amputation, tagged LTD is reduced or completely blocked. Our results strongly suggest that brain cortices of adult mice are highly plastic and show biphasic tagging of plasticity. These findings of tagged LTD may provide a new direction for future treatment of phantom pain and amputation-related emotional disorders.

Enhanced long-term potentiation in the anterior cingulate cortex of tree shrew.

Synaptic plasticity is a key cellular model for learning, memory and chronic pain. Most previous studies were carried out in rats and mice, and less is known about synaptic plasticity in non-human primates. In the present study, we used integrative experimental approaches to study long-term potentiation (LTP) in the anterior cingulate cortex (ACC) of adult tree shrews. We found that glutamate is the major excitatory transmitter and α-amino-3-hydroxy-5-methyl-4-isoxazole-propionicacid (AMPA) receptors mediate postsynaptic responses. LTP in tree shrews was greater than that in adult mice and lasted for at least 5 h. N-methyl-d-aspartic acid (NMDA) receptors, Ca2+ influx and adenylyl cyclase 1 (AC1) contributed to tree shrew LTP. Our results suggest that LTP is a major form of synaptic plasticity in the ACC of primate-like animals. This article is part of a discussion meeting issue 'Long-term potentiation: 50 years on'.

Cortical nitric oxide required for presynaptic long-term potentiation in the insular cortex.

Nitric oxide (NO) is a key diffusible messenger in the mammalian brain. It has been proposed that NO may diffuse retrogradely into presynaptic terminals, contributing to the induction of hippocampal long-term potentiation (LTP). Here, we present novel evidence that NO is required for kainate receptor (KAR)-dependent presynaptic form of LTP (pre-LTP) in the adult insular cortex (IC). In the IC, we found that inhibition of NO synthase erased the maintenance of pre-LTP, while the induction of pre-LTP required the activation of KAR. Furthermore, NO is essential for pre-LTP induced between two pyramidal cells in the IC using the double patch-clamp recording. These results suggest that NO is required for homosynaptic pre-LTP in the IC. Our results present strong evidence for the critical roles of NO in pre-LTP in the IC. This article is part of a discussion meeting issue 'Long-term potentiation: 50 years on'.

Inhibition of cortical synaptic transmission, behavioral nociceptive, and anxiodepressive-like responses by arecoline in adult mice.

Areca nut, the seed of Areca catechu L., is one of the most widely consumed addictive substances in the world after nicotine, ethanol, and caffeine. The major effective constituent of A. catechu, arecoline, has been reported to affect the central nervous system. Less is known if it may affect pain and its related emotional responses. In this study, we found that oral application of arecoline alleviated the inflammatory pain and its induced anxiolytic and anti-depressive-like behavior. Arecoline also increased the mechanical nociceptive threshold and alleviated depression-like behavior in naïve mice. In the anterior cingulate cortex (ACC), which acts as a hinge of nociception and its related anxiety and depression, by using the multi-electrode field potential recording and whole-cell patch-clamp recording, we found that the evoked postsynaptic transmission in the ACC of adult mice has been inhibited by the application of arecoline. The muscarinic receptor is the major receptor of the arecoline in the ACC. Our results suggest that arecoline alleviates pain, anxiety, and depression-like behavior in both physiological and pathological conditions, and this new mechanism may help to treat patients with chronic pain and its related anxiety and disorder in the future.

Inhibition of calcium-stimulated adenylyl cyclase subtype 1 (AC1) for the treatment of pain and anxiety symptoms in Parkinson's disease mice model.

Pain and anxiety are two common and undertreated non-motor symptoms in Parkinson's disease (PD), which affect the life quality of PD patients, and the underlying mechanisms remain unclear. As an important subtype of adenylyl cyclases (ACs), adenylyl cyclase subtype 1 (AC1) is critical for the induction of cortical long-term potentiation (LTP) and injury induced synaptic potentiation in the cortical areas including anterior cingulate cortex (ACC) and insular cortex (IC). Genetic deletion of AC1 or pharmacological inhibition of AC1 improved chronic pain and anxiety in different animal models. In this study, we proved the motor deficit, pain and anxiety symptoms of PD in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated mice model. As a lead candidate AC1 inhibitor, oral administration (1 dose and seven doses) of NB001 (20 and 40 mg/kg) showed significant analgesic effect in MPTP-treated mice, and the anxiety behavior was also reduced (40 mg/kg). By using genetic knockout mice, we found that AC1 knockout mice showed reduced pain and anxiety symptoms after MPTP administration, but not AC8 knockout mice. In summary, genetic deletion of AC1 or pharmacological inhibition of AC1 improved pain and anxiety symptoms in PD model mice, but didn't affect motor function. These results suggest that NB001 is a potential drug for the treatment of pain and anxiety symptoms in PD patients by inhibiting AC1 target.

The Pathway-Selective Dependence of Nitric Oxide for Long-Term Potentiation in the Anterior Cingulate Cortex of Adult Mice.

Nitric oxide (NO) is a key diffusible messenger in the mammalian brain. It has been proposed that NO may diffuse in retrograde into presynaptic terminals, contributing to the induction of hippocampal long-term potentiation (LTP). Here, we present novel evidence that NO is selectively required for the synaptic potentiation of the interhemispheric projection in the anterior cingulate cortex (ACC). Unilateral low-frequency stimulation (LFS) induced a short-term synaptic potentiation on the contralateral ACC through the corpus callosum (CC). The use of the antagonists of the NMDA receptor (NMDAR), or the inhibitor of the L-type voltage-dependent Ca2+ channels (L-VDCCs), blocked the induction of this ACC-ACC potentiation. In addition, the inhibitor of NO synthase, or inhibitors for its downstream signaling pathway, also blocked this ACC-ACC potentiation. However, the application of the NOS inhibitor blocked neither the local electric stimulation-induced LTP nor the stimulation-induced recruitment of silent responses. Our results present strong evidence for the pathway-selective roles of NO in the LTP of the ACC.

Alleviation of migraine related pain and anxiety by inhibiting calcium-stimulating AC1-dependent CGRP in the insula of adult rats.

BACKGROUND: Recent animal and clinical findings consistently highlight the critical role of calcitonin gene-related peptide (CGRP) in chronic migraine (CM) and related emotional responses. CGRP antibodies and receptor antagonists have been approved for CM treatment. However, the underlying CGRP-related signaling pathways in the pain-related cortex remain poorly understood. METHODS: The SD rats were used to establish the CM model by dural infusions of inflammatory soup. Periorbital mechanical thresholds were assessed using von-Frey filaments, and anxiety-like behaviors were observed via open field and elevated plus maze tests. Expression of c-Fos, CGRP and NMDA GluN2B receptors was detected using immunofluorescence and western blotting analyses. The excitatory synaptic transmission was detected by whole-cell patch-clamp recording. A human-used adenylate cyclase 1 (AC1) inhibitor, hNB001, was applied via insula stereotaxic and intraperitoneal injections in CM rats. RESULTS: The insular cortex (IC) was activated in the migraine model rats. Glutamate-mediated excitatory transmission and NMDA GluN2B receptors in the IC were potentiated. CGRP levels in the IC significantly increased during nociceptive and anxiety-like activities. Locally applied hNB001 in the IC or intraperitoneally alleviated periorbital mechanical thresholds and anxiety behaviors in migraine rats. Furthermore, CGRP expression in the IC decreased after the hNB001 application. CONCLUSIONS: Our study indicated that AC1-dependent IC plasticity contributes to migraine and AC1 may be a promising target for treating migraine in the future.

Recruitment of cortical silent responses by forskolin in the anterior cingulate cortex of adult mice.

Recent studies using different experimental approaches demonstrate that silent synapses may exist in the adult cortex including the sensory cortex and anterior cingulate cortex (ACC). The postsynaptic form of long-term potentiation (LTP) in the ACC recruits some of these silent synapses and the activity of calcium-stimulated adenylyl cyclases (ACs) is required for such recruitment. It is unknown if the chemical activation of ACs may recruit silent synapses. In this study, we found that activation of ACs contributed to synaptic potentiation in the ACC of adult mice. Forskolin, a selective activator of ACs, recruited silent responses in the ACC of adult mice. The recruitment was long-lasting. Interestingly, the effect of forskolin was not universal, some silent synapses did not undergo potentiation or recruitment. These findings suggest that these adult cortical synapses are not homogenous. The application of a selective calcium-permeable AMPA receptor inhibitor 1-naphthyl acetyl spermine (NASPM) reversed the potentiation and the recruitment of silent responses, indicating that the AMPA receptor is required. Our results strongly suggest that the AC-dependent postsynaptic AMPA receptor contributes to the recruitment of silent responses at cortical LTP.

Anterior cingulate cortex and insomnia: A cingulate-striatum connection.

Insomnia is an important comorbidity of chronic pain. In this issue of Neuron, Li et al. report that chronic-pain-induced insomnia is mediated by the pyramidal neurons in the anterior cingulate cortex and their dopaminergic projections to the dorsal medial striatum.

Fear memory regulation by the cAMP signaling pathway as an index of reexperiencing symptoms in posttraumatic stress disorder.

Posttraumatic stress disorder (PTSD) is a psychiatric disorder associated with traumatic memory, yet its etiology remains unclear. Reexperiencing symptoms are specific to PTSD compared to other anxiety-related disorders. Importantly, reexperiencing can be mimicked by retrieval-related events of fear memory in animal models of traumatic memory. Recent studies revealed candidate PTSD-associated genes that were related to the cyclic adenosine monophosphate (cAMP) signaling pathway. Here, we demonstrate the tight linkage between facilitated cAMP signaling and PTSD by analyzing loss- and gain-of-cAMP signaling effects on fear memory in mice and the transcriptomes of fear memory-activated mice and female PTSD patients with reexperiencing symptoms. Pharmacological and optogenetic upregulation or downregulation of cAMP signaling transduction enhanced or impaired, respectively, the retrieval and subsequent maintenance of fear memory in mice. In line with these observations, integrative mouse and human transcriptome analysis revealed the reduced mRNA expression of phosphodiesterase 4B (PDE4B), an enzyme that degrades cAMP, in the peripheral blood of PTSD patients showing more severe reexperiencing symptoms and the mouse hippocampus after fear memory retrieval. Importantly, more severe reexperiencing symptoms and lower PDE4B mRNA levels were correlated with decreased DNA methylation of a locus within PDE4B, suggesting the involvement of methylation in the mechanism of PTSD. These findings raise the possibility that the facilitation of cAMP signaling mediating the downregulation of PDE4B expression enhances traumatic memory, thereby playing a key role in the reexperiencing symptoms of PTSD patients as a functional index of these symptoms.

Selective enhancement of fear extinction by inhibiting neuronal adenylyl cyclase 1 (AC1) in aged mice.

Adenylyl cyclase 1 (AC1) is a selective subtype of ACs, which is selectively expressed in neurons. The activation of AC1 is activity-dependent, and AC1 plays an important role in cortical excitation that contributes to chronic pain and related emotional disorders. Previous studies have reported that human-used NB001 (hNB001, a selective AC1 inhibitor) produced analgesic effects in different animal models of chronic pain. However, the potential effects of hNB001 on learning and memory have been less investigated. In the present study, we found that hNB001 affected neither the induction nor the expression of trace fear, but selectively enhanced the relearning ability during the extinction in aged mice. By contrast, the same application of hNB001 did not affect recent, remote auditory fear memory, or remote fear extinction in either adult or aged mice. Furthermore, a single or consecutive 30-day oral administration of hNB001 did not affect acute nociceptive response, motor function, or anxiety-like behavior in either adult or aged mice. Our results are consistent with previous findings that inhibition of AC1 did not affect general sensory, emotional, and motor functions in adult mice, and provide strong evidence that inhibiting the activity of AC1 may be beneficial for certain forms of learning and memory in aged mice.

Long-term plasticity of NMDA GluN2B (NR2B) receptor in anterior cingulate cortical synapses.

The anterior cingulate cortex (ACC) is a key cortical area for pain perception, emotional fear and anxiety. Cortical excitation is thought to be the major mechanism for chronic pain and its related emotional disorders such as anxiety and depression. GluN2B (or called NR2B) containing NMDA receptors play critical roles for such excitation. Not only does the activation of GluN2B contributes to the induction of the postsynaptic form of LTP (post-LTP), long-term upregulation of GluN2B subunits through tyrosine phosphorylation were also detected after peripheral injury. In addition, it has been reported that presynaptic NMDA receptors may contribute to the modulation of the release of glutamate from presynaptic terminals in the ACC. It is believed that inhibiting subtypes of NMDA receptors and/or downstream signaling proteins may serve as a novel therapeutic mechanism for future treatment of chronic pain, anxiety, and depression.

Treatment and prognosis analysis of 488 patients with FIGO 2018 stage Ⅲc squamous cervical cancer / 中华妇产科杂志

Objective: To analyze the treatment and prognosis of patients with International Federation of Gynecology and Obstetrics (FIGO) 2018 stage Ⅲc cervical squamous cell carcinoma. Methods: A total of 488 patients at Zhejiang Cancer Hospital between May, 2013 to May, 2015 were enrolled. The clinical characteristics and prognosis were compared according to the treatment mode (surgery combined with postoperative chemoradiotherapy vs radical concurrent chemoradiotherapy). The median follow-up time was (96±12) months ( range time from 84 to 108 months). Results: (1) The data were divided into surgery combined with chemoradiotherapy group (surgery group) and concurrent chemoradiotherapy group (radiotherapy group), including 324 cases in the surgery group and 164 cases in the radiotherapy group. There were significant differences in Eastern Cooperation Oncology Group (ECOG) score, FIGO 2018 stage, large tumors (≥4 cm), total treatment time and total treatment cost between the two groups (all P<0.01). (2) Prognosis: ① for stage Ⅲc1 patients, there were 299 patients in the surgery group with 250 patients survived (83.6%). In the radiotherapy group, 74 patients survived (52.9%). The difference of survival rates between the two groups was statistically significant (P<0.001). For stage Ⅲc2 patients, there were 25 patients in surgery group with 12 patients survived (48.0%). In the radiotherapy group, there were 24 cases, 8 cases survived, the survival rate was 33.3%. There was no significant difference between the two groups (P=0.296). ② For patients with large tumors (≥4 cm) in the surgery group, there were 138 patients in the Ⅲc1 group with 112 patients survived (81.2%); in the radiotherapy group, there were 108 cases with 56 cases survived (51.9%). The difference between the two groups was statistically significant (P<0.001). Large tumors accounted for 46.2% (138/299) vs 77.1% (108/140) in the surgery group and radiotherapy group. The difference between the two groups was statistically significant (P<0.001). Further stratified analysis, a total of 46 patients with large tumors of FIGO 2009 stage Ⅱb in the radiotherapy group were extracted, and the survival rate was 67.4%, there was no significant difference compared with the surgery group (81.2%; P=0.052). ③ Of 126 patients with common iliac lymph node, 83 patients survived, with a survival rate of 65.9% (83/126). In the surgery group, 48 patients survived and 17 died, with a survival rate of 73.8%. In the radiotherapy group, 35 patients survived and 26 died, with a survival rate of 57.4%. There were no significant difference between the two groups (P=0.051). (3) Side effects: the incidence of lymphocysts and intestinal obstruction in the surgery group were higher than those in the radiotherapy group, and the incidence of ureteral obstruction and acute and chronic radiation enteritis were lower than those in the radiotherapy group, and there were statistically significant differences (all P<0.01). Conclusions: For stage Ⅲc1 patients who meet the conditions for surgery, surgery combined with postoperative adjuvant chemoradiotherapy and radical chemoradiotherapy are acceptable treatment methods regardless of pelvic lymph node metastasis (excluding common iliac lymph node metastasis), even if the maximum diameter of the tumor is ≥4 cm. For patients with common iliac lymph node metastasis and stage Ⅲc2, there is no significant difference in the survival rate between the two treatment methods. Based on the duration of treatment and economic considerations, concurrent chemoradiotherapy is recommended for the patients.

Efficacy of ultra-mini percutaneous nephrolithotomy and retrograde intrarenal surgery in the treatment of 2-3 cm lower calyceal stones / Eficacia de la nefrolitotomía percutánea ultramini y la cirugía intrarrenal retrógrada en el tratamiento de cálculos caliceales inferiores de 2-3 cm

Abstract We aimed to compare the efficacy and safety of ultra-mini percutaneous nephrolithotomy (UMP) and retrograde intrarenal surgery (RIRS) for the management of lower calyceal stones. A group of 136 patients with a single lower calyceal stone (2-3 cm in diameter) was divided into the UMP or RIRS groups. The average operation time in the RIRS group was significantly longer than that in the UMP group, and the intraoperative blood loss in the former was markedly less than that in the latter. Besides, in the RIRS group, the decreased value of postoperative Hb was obviously lower, the postoperative hospital stay was evidently shorter, and the total hospitalization expenses were markedly less than those in UMP group were. Moreover, the success rate of the first-stage lithotripsy in the UMP group was notably higher than that in RIRS group. The RIRS group had an obviously lower VAS score but a markedly higher BCS score than the UMP group six hours after surgery. At 24 h after operation, the levels of serum CRP, TNF-α and IL -6 in patients in both groups were remarkably increased, and they were evidently lower in the RIRS group than those in the UMP group were. Three days after surgery, the levels of serum CRP, TNF-α and IL -6 were notably lower in the UMP group than those in RIRS group were. RIRS and UMP are safe and effective in the treatment of 2-3 cm lower calyceal stones. The first-stage UMP is characterized by a high stone-free rate (SFR), short operation time and low postoperative infection risk, while RIRS is associated with less blood loss and low total expenses.

Resumen Nuestro objetivo fue comparar la eficacia y seguridad de la nefrolitotomía percutánea ultramini (UMP) y la cirugía intrarrenal retrógrada (CRIR) en el manejo quirúrgico de los cálculos caliceales inferiores. Un grupo de 136 pacientes con un solo cálculo calicial inferior (2-3 cm de diámetro) se dividió en un grupo UMP o un grupo CRIR. El tiempo de operación promedio en el grupo CRIR fue significativamente más largo que en el grupo UMP, y la pérdida de sangre intraoperatoria en el primero fue marcadamente menor que en el segundo. Además, en el grupo CRIR, el valor disminuido de la Hb postoperatoria fue obviamente menor, la estancia hospitalaria postoperatoria fue evidentemente más corta y los gastos totales de hospitalización fueron notablemente menores que los del grupo UMP. Además, la tasa de éxito de la litotricia de primera etapa en el grupo UMP fue notablemente más alta que en el grupo CRIR. El grupo CRIR tuvo una puntuación VAS obviamente más baja pero una puntuación BCS marcadamente más alta que el grupo UMP a seos horas después de la operación. A las 24 h después de la operación, los niveles séricos de PCR, TNF-α e IL -6 en los pacientes de ambos grupos aumentaron notablemente y fueron evidentemente más bajos en el grupo CRIR que en el grupo UMP. Tres días después de la operación, los niveles séricos de PCR, TNF-α e IL -6 fueron notablemente más bajos en el grupo UMP que en el grupo CRIR. Los procedimientos CRIR y el UMP son seguros y eficaces en el tratamiento de cálculos caliciales inferiores de 2-3 cm. El UMP de primera etapa se caracteriza por tener una tasa libre de cálculo (SFR) alta, un tiempo de operación corto y un riesgo de infección posoperatorio bajo, y el RIRS se caracteriza por una menor pérdida de sangre y gastos totales bajos.

Tuning a bi-enzymatic cascade reaction in Escherichia coli to facilitate NADPH regeneration for ε-caprolactone production.

ε-Caprolactone is a monomer of poly(ε-caprolactone) which has been widely used in tissue engineering due to its biodegradability and biocompatibility. To meet the massive demand for this monomer, an efficient whole-cell biocatalytic approach was constructed to boost the ε-caprolactone production using cyclohexanol as substrate. Combining an alcohol dehydrogenase (ADH) with a cyclohexanone monooxygenase (CHMO) in Escherichia coli, a self-sufficient NADPH-cofactor regeneration system was obtained. Furthermore, some improved variants with the better substrate tolerance and higher catalytic ability to ε-caprolactone production were designed by regulating the ribosome binding sites. The best mutant strain exhibited an ε-caprolactone yield of 0.80 mol/mol using 60 mM cyclohexanol as substrate, while the starting strain only got a conversion of 0.38 mol/mol when 20 mM cyclohexanol was supplemented. The engineered whole-cell biocatalyst was used in four sequential batches to achieve a production of 126 mM ε-caprolactone with a high molar yield of 0.78 mol/mol.

Prognostic thirteen-long non-coding RNAs (IncRNAs) could improve the survival prediction of gastric cancer / El pronóstico de 13 largos RNA no codificantes (IncRNA) podría mejorar la predicción de supervivencia del cáncer gástrico

OBJECTIVE: Accumulating evidence has demonstrated that long non-coding RNAs (lncRNAs) play important regulatory roles in the tumorigenesis and progression of gastric cancer (GC). The aim of this study was to construct the prognostic predictive model of lncRNAs signature and improve the survival prediction of GC. PATIENTS AND METHODS: The expression profiling of lncRNAs in large GC cohorts was performed from The Cancer Genome Atlas (TCGA) databases using the lncRNAs-mining approach, including training data set (N=160) and testing data set (N=159). A 13-lncRNAs signature significantly associated with overall survival (OS) in the training data set was selected. The prognostic value of this 13-lncRNAs signature was then confirmed in the test validation set and the entire validation set, respectively. RESULTS: Based on lncRNA expression profiling of 319 patients with stomach adenocarcinoma (STAD), prognostic 13-lncRNAs signature was found to be significantly associated with the prognosis of GC. Compared to patients with low-risk scores, patients with high-risk scores had a significantly shorter survival time. Moreover, functional enrichment analysis indicated that this 13-lncRNAs signature was potentially involved in multiple biological processes, such as DNA replication and cell cycle signaling pathway. CONCLUSIONS: The prognostic model of the 13-lncRNAs signature established by our study could improve the survival prediction of GC to a greater extent

OBJETIVO: Las pruebas acumuladas demostraron que los ARN no codificantes de larga duración (ARNlC) desempeñaban los importantes papeles reguladores en la tumorigénesis y la progresión del cáncer gástrico (CG). El objetivo de este estudio fue construir el modelo predictivo de pronóstico de la firma de los lncRNA y mejorar la predicción de supervivencia del GC. PACIENTES Y MÉTODOS: El perfil de expresión de los lncARN en grandes cohortes de GC se realizó a partir de las bases de datos del Atlas del Genoma del Cáncer (TCGA) utilizando el enfoque de minería de lncARN, incluyendo el conjunto de datos de entrenamiento (N=160) y el conjunto de datos de pruebas (N=159). Se eligió la firma de 13 lncARN significativamente asociada con la supervivencia general (OS) en la serie de capacitación. El valor pronóstico de esta firma de 13-lncARN se confirmó luego en la serie de validación de pruebas y en toda la serie de validación, respectivamente. RESULTADOS: Basado en el perfil de expresión de lncRNA de 319 pacientes con adenocarcinoma de estómago (STAD), se encontró que la firma de 13-lncRNA de pronóstico estaba significativamente asociada con el pronóstico de GC. En comparación con los pacientes con puntuaciones de bajo riesgo, los pacientes con puntuaciones de alto riesgo tuvieron un tiempo de supervivencia significativamente más corto. Además, el análisis de enriquecimiento funcional indicó que esta firma de 13-lncARN estaba potencialmente involucrada en múltiples procesos biológicos, como la replicación del ADN y la vía de señalización del ciclo celular. CONCLUSIONES: El modelo de pronóstico de la firma de 13-lncARN establecido por nuestro estudio podría mejorar mejor la predicción de supervivencia del GC

Effects of serum starvation on cell cycle synchronization in primary human umbilical vein endothelial cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the optimal starvation conditions of human umbilical vein endothelial cells (HUVECs) and establish a highly efficient and stable method for separating HUVECs.</p><p><b>METHODS</b>HUVECs harvested from human umbilical cords by digestion with 0.1% collagenase II for 15 min were cultured in endothelial culture medium (ECM) containing 5% fetal bovine serum (FBS), 1% endothelial cell growth factor (ECGS) and 1% penicillin/streptomycin solution(P/S) at 37 degrees celsius; in 5% CO2. The cells were observed for cell morphology under an inverted microscope and identified with immunofluorescence assay. The purity of HUVECs was detected using flow cytometry (FCM). The cell cycles of HUVECs cultured in the presence of 0, 0.1%, 0.5%, and 1% FBS for 0, 6, 12, 18, and 24 h were analyzed with flow cytometry.</p><p><b>RESULTS</b>s The purity of HUVECs harvested by digestion with 0.1% collagenase II reached 99.67%. The primary HUVECs showed a cobblestone or volute appearance in vitro. Immunocytochemistry showed that HUVECs highly expressed VIII-related antigen. Cell culture in the presence of different concentrations of FBS for 6 h resulted in 70% G0/G1 phase cells, which increased to 80%-90% at 12 h of cell culture, and further to around 95% at 18 and 24 h.</p><p><b>CONCLUSION</b>Digestion with 0.1% collagenase II can obtain high-purity primary HUVECs. Culturing HUVECs in serum-free medium for 12 h can result in a high purity (over 80%) of G0/G1 phase cells.</p>

Effect of reinforcing qi for resolving masses method on expressions of MEK2 and p-ERK protein in estrogen induced uterine leiomyoma model guinea pigs' uterine tissue / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the effect of reinforcing qi for resolving masses method (RQRMM) on expressions of extracellular signal regulated kinase 2 (MEK2) and phosphorylation extracellular signal regulated kinase (p-ERK) protein in estrogen induced uterine leiomyoma model Guinea pigs' uterine tissue.</p><p><b>METHODS</b>Guinea pigs were randomly divided into five groups, i.e., the model group, the high dose group, the middle dose group, the low dose group, and the Western medicine group (mifepristone). The normal control group was set up. The uterine leiomyoma model in guinea pigs was established by castrating and subcutaneous injecting estradiol (E2). The protein expression levels of MEK2 and p-ERK of guinea pigs' uterine tissues were detected by immunohistochemical assay.</p><p><b>RESULTS</b>The protein expressions of MEK2 and p-ERK in the uterine muscular tissue of Guinea pigs' uterine tissue were higher in the model group than in the normal group (P < 0.01). They decreased to some degree in the high dose group, the middle dose group, and the low dose group. Of them, the protein expressions of MEK2 and p-ERK were significantly lower in the high dose group than in the model group and the Western medicine group (P < 0.01).</p><p><b>CONCLUSION</b>RQRMM could treat uterine leiomyoma possibly through intervening the MAPK/ERK cell signal pathway to inhibit the proliferation of myoma cells.</p>

Investigate circulating levels of chemokines and evaluate the correlation between these chemokines and liver function indicators in autoimmune hepatitis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>This study investigated circulation levels of chemokines (CCL2, CCL5, CXCL8, CXCL9, CXCL10) in autoimmune hepatitis(AIH) patients and evaluated the correlation between these chemokines and liver function indicators.</p><p><b>METHODS</b>A total of 5 chemokines (CCL2, CCL5, CXCL8, CXCL9, CXCL10) were measured simultaneously by cytokine beads assay(CBA) in the sera of 46 patients with AIH and 12 cases of healthy control.</p><p><b>RESULTS</b>In this study we found that serum levels of CCL2 , CXCL9 and CXCL10 in AIH patients and healthy controls were 11.79:8.39 pg/ml, 11.31:2.69 pg/ml, 15.85:4.64 pg/ml, respectively , which implied these chemokines were significantly higher in AIH patients when compared to healthy control (Z=-1.958, P=0.05; Z=-4.527, P less than 0.0001; Z=-3.84, P less than 0.0001, respectively). And circulation levels of CCL2 , CXCL8 , CXCL9 and CXCL10 in pretreatment and remission stages of patients with AIH were 29.69:11.16 pg/ml, 7.2:5.38 pg/ml, 16.02:5.47 pg/ml, 90.01:13.24 pg/ml, respectively, which showed these chemokines decreased during remission from pretreatment stage levels (t=2.985, P=0.005; Z=-2.547, P=0.0112; Z=-3.187, P=0.001; t=2.12, P=0.0015, respectively). Among AIH , CXCL8 was correlated positively with lgG(r2=0.291, P=0.0039); CXCL9 was associated positively with ALT and AST(r2=0.5324 , P less than 0.0001; r2=0.3352, P less than 0.0001); CXCL10 showed a positive correlation with ALT , AST and GGT(r2=0.9551, P less than 0.0001; r2=0.8960, P less than 0.0001; r2=0.8271, P less than 0.0001).</p><p><b>CONCLUSION</b>Serum levels of CCL2, CXCL8, CXCL9 and CXCL10 are significantly higher in patients with AIH, but decrease to levels in healthy controls after successful treatment , and circulation levels of CXCL9 and CXCL10 are associated positively with liver function indicators which can react inflammation activity of liver, all these may imply that chemokines can reflect the degree of liver inflammation and may be one of the main culprits in AIH pathological damage.</p>