Evaluation and Improvement of Pumping Well Operating Conditions in an Oil Field Block Based on Grey Correlation Analysis

The "Oil and Gas Water Well Production Data Management System Database" provides great assistance for oilfield production, monitoring, and management. However, due to the harsh conditions of oil field wells and the lack of some test data, traditional management methods are no longer suitable for present condition. At the same time, optimization analysis for a single oil well has a high cost and low efficiency, and it is difficult to achieve the modern management goal of large-scale pumping well groups. In this paper, the grey correlation method is used to analyze the direct correlation between the influencing factors and the system efficiency, surface equipment driving efficiency, and wellbore lifting efficiency, and the improvement method against factors with strong correlation is prioritized. A multi-node evaluation index system for pumping well systems and corresponding improvement methods were constructed, and evaluation software was compiled. This technology considers the running condition of the pumping unit in one oil field block, and selects the oil wells to be improved according to the evaluation index, and puts forward the targeted improvement methods according to the common problems of the oil well. This paper provides a set of reliable technical methods for the efficient management of the oil well in the oil field block

PO-305 An 8-week, low carbohydrate, high fat, ketogenic diet enhanced exercise capacity through improved ketolysis and lipolysis in mice

Objective Carbohydrates, lipids and proteins are utilized both for energy production and structure of body. Among them, protein is the most important component of our body, carbohydrates and lipids are more flexible for energy supply system. Due to carbohydrate pitfall and lipid reserve abundance, coaches and elite athletes aspirate for an effective way to enhance fat utilization. Meanwhile, intramuscular triacylglycerol (IMTG) is a special way for skeletal muscle to store lipids. During exercise, IMTG may contribute up to 20% of total energy turnover, thus contribute significantly for ATP synthesis during exercise. However, abnormal or excessive fat deposition in skeletal muscle may induce insulin resistance as well. Intramuscular lipolysis regulation is crucial for energy supply system during exercise. It is reported by Amati and colleagues that well-trained athletes exhibit higher levels of IMTG and diacylglycerol (DAG) as well as well-preserved sensitivity to insulin, indicating lipolysis ability may be enhanced during exercise. In our previous study, we reported that an 8-week, a low carbohydrate, ketogenic diet increased running time till exhausted in male C57BL6/J mice, presuming the mechanism to be enhanced fat utilization. In the present study, we observed the alternation pattern of messenger RNAs related to lipid mobilization, fatty acid utilization and ketone body oxidation in muscle and adipose tissue immediately after exercise in both Type 1 and Type 2 muscle fibers. Materials and Methods Male C57BL/6J mice (n = 35) were purchased from Takasugi Experimental Animals Supply (Kasukabe, Japan) at 7 weeks of age. All mice were randomly divided into four groups: chow diet (control: Con), including chow diet, sedentary (n = 8) and chow diet plus exercise (Con + Ex, n = 9), ketogenic diet (KD), including KD, sedentary, n = 9, and KD plus exercise (KD + Ex, n = 9) groups. A KD diet TP-201450 (consisting of 76.1% fat, 8.9% protein and 3.5% carbohydrate, 7.342 kcal/g) and a chow diet AIN93G (consisting of 7% fat, 17.8% protein and 64.3% carbohydrate, 3.601 kcal/g) wt/wt were obtained from Trophic (TROPHIC Animal Feed High-tech Co., Ltd., Nantong, Jiangsu, China). Mice were maintained on ad libitum chow diet or KD. Total RNA was extracted from the gastrocnemius muscle, soleus muscle and epididymal adipose tissue using the RNeasy Mini Kit or RNeasy Lipid Mini Kit (Qiagen, Valencia, CA) according to the manufacturer’s instructions. Total RNA was reverse transcribed to cDNA using the High Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA) according to the manufacturer’s instructions. PCR was performed with the Fast 7500 real-time PCR system (Applied Biosystems) using the Fast SYBR® Green PCR Master Mix (Applied Biosystems). Plasma IL-6 was measured using a R&D Mouse ELISA Duo set (R&D Systems, Minneapolis, USA) according to the manufacture’s instructions. Plasma glycerol was measured using Glycerol Colorimetric Assay Kit (Cayman Chemical Co., Ann Arbor, MI, USA). Results and Discussion 3.1 IL-6 concentration and exercise-induced myokine IL-6 mRNA alternation in both muscle fiber IL-6 plays essential roles in immune responses. However, exercise induced IL-6 is reported to be able to stimulate lipolysis both in IMTG pool intramuscularly and adipocytes. Defined as exercise factor, or so-called myokines, muscle-derived IL-6 exhibits regulating function in various experiment circumstances. Recombinant human IL-6 infusion showed an enhanced lipolysis and fat oxidation capacity in human subjects. Genetically IL-6 deficient mice presented a reduced ability on lipolysis and fatty acid oxidation. During KD administration, fat oxidation is no doubt the predominant, if not the only origin of energy, this make us to suspect IL-6 may be altered by acute exercise. As shown in Figure 1, IL-6 mRNA increased rapidly, with a nearly 100-fold change in slow-twitch muscle fiber, and KD helped to this up-regulation. Transcription IL-6 level increased significantly in KD, compared to Con group subject, in soleus muscle, under the context of exhaustive exercise. This makes us to suspect that IL-6 may contribute to enhanced lipolysis and fatty acid mobilization. However, the effect is not observed in fast-twitch muscle. The result indicated that IL-6 mRNA expression exhibited a muscle fiber specification. Slow-twitch muscle fiber contributes more to endurance exercise, as fast-twitch muscle fiber mainly contributes to explosive strength and acceleration. The difference of fiber function leads to a different secretion mode of IL-6. The error bar is high in both exercise group. We observed an interesting phenomenon: the mice who quit at around 200 minutes has the highest IL-6 mRNA expression (gastrocnemius muscle) and plasma IL-6 concentration in both groups. One reason may be that, as the exercise taking on and gradually reach final fatigue, the call for fatty acid decreases with time. As shown in Figure 2, both muscular IL-6 protein and plasma IL-6 were increased by exhaustive exercise. However, plasma IL-6 is significantly lower in the KD plus exercise group, though IL-6 rose nearly 5-fold in control feed group, it only rose to 2.5-fold in the KD group after exercise.  For this phenomenon, the best explanation is that the well-adaption of lipid-centered metabolism, including metabolic flexibility and increased IMTG reservoir weakened the need to pull the trigger; this may also be the answer why KD mice had lower weight. 3.2 Fatty acid mobilation related RNA alternation after exhaustive exercise under endogenous ketosis in epididymal adipose tissue Adipose triglyceride lipase (ATGL) is also known as desnutrin in the first place, is a kind of lipase whose substance is patatin-like phospholipase domain-containing protein. Hormone-sensitive lipase (HSL) is also known as cholesteryl ester hydrolase (CEH), is another intracellular neutral lipase. ATGL and HSL cooperated to break apart fatty acids from TG, after which IMTG-origin fatty acid will be directly used for beta-oxidation, or lipid drop-origin fatty acids will be transported though lipoprotein shipping in the form of VLDL from adipose tissue into muscle fibers during exercise. As shown in Figure 3, mRNA expression levels of lipase were significantly enhanced by KD or exercise, indicating the up-regulated lipid mobilization and utilization ability is enhanced by exercise in adipocytes. However, KD plus exercise reversed this increase. One plausible explain for this phenomenon is the lack of plasma IL-6, thus the ability to mobilize fatty acid from adipose tissue is reduced. Adrenergic blocking agents are reported to harm fatty acid mobilization during fasting, and IL-6 is reported to function as adrenergic hormone. Adipocyte-specific HSL deficiency mice present lowered submaximal exercise capacity. Our experiment design, the protocol for treadmill running is similar to a submaximal exercise. Under this circumstance, fat mobilization seems to be critical. Loss of this mobilizing ability, while exercise capacity is yet enhanced, makes us to suspect whether IMTG plays dominant role in this process. 3.3 Ketolytic RNA alternation after exhaustive exercise under conditions of endogenous ketosis in Type 1 and Type 2 muscle fiber Ketolysis is a complete oxidation of ketone bodies. Ketone bodies are utilized by mitochondria of extrahepatic tissues via a series of enzymatic reactions. Ketolysis is regulated by a rate-limiting enzyme 3-oxoacid CoA-transferase 1 (OXCT)-1 and hydroxybutyrate dehydrogenase (HBDH). Thus, we measured the transcriptional alternation of these enzymes in different muscle tissues. In our previous study, plasma ketone body (KB) increased rapidly in the sedentary KD group. However, after exhaustive exercise, blood KB of those KD mice dropped dramatically, while situation of blood KD in the Con mice showed a different figure. These results indicated that 1n 8-week KD administration has improved ketolysis, the ability for subjects to utilize KB. To investigate the mechanism of this enhancement, we assessed key enzymes in ketolysis in both fiber types. As shown in Figure 4, gene expressions of these enzymes also present a fiber-specification. Since fast-twitch muscle fiber plays a second role in endurance exercise, exercise did not alter ketolytic enzymes in the transcriptional level, in gastrocnemius tissue. However, in the slow-twitch muscle fiber, it was changed. HBDH is up-regulated significantly in the case of KD plus exercise. Results here indicated that HBDH plays the key role in the improvement of exercise capacity by an 8-week KD. 3.4 Lipolysis- and fatty acid oxidation related RNA alternation after exhaustive exercise during endogenous ketosis in Type 1 and Type 2muscle fiber After reaching working site, muscle lipoprotein lipase (LPL) hydrolyzes VLDL and harvests fatty acids at last, which will be finally utilized as primary fuel. Carnitine palmitoyl transferase (CPT)-1A, acyl-CoA oxidase (ACO), hydroxyacyl-coenzyme A dehydrogenase (HADH), medium chain acyl-CoA dehydrogenase (MCAD) and malonyl-CoA decarboxylase (MCD) are key regulating enzymes during fatty acid beta-oxidation. And for IMTG, the fatty acid is harvest by intramuscular lipase, ATGL and HSL. In an article published several years ago, the authors called adipose ATGL and HSL, “the mover and shaker of muscle lipolysis”. As shown in Figure 5, both ATGL and HSL mRNA expression are up-regulated by KD, but not by exhaustive exercise. Combined with results in the above part, enhanced mobilization of fatty acid intramuscularly is the main factor, but not the mobilization from adipocyte. LPL mRNA expression in gastrocnemius exhibited the same pattern of lipase mRNA synthesized by adipose tissue. Combined together, the reduced fatty mobilization from adipocyte, partly being the results of higher blood NEFA and TG, accompanied with enhanced fatty acid gain from IMTG pool, need for LPL was reduced.  As shown in Figure 6, in fast-twitch muscle fiber, CPT-1a, ACO and HADH mRNA expressions are enhanced by KD during exhaustive exercise. In slow-twitch muscle fiber, feed played as a main factor regulating fatty acid oxidation. CPT1a, MCAD and MCD mRNA expressions are enhanced. In summary, though tissue specific specificity were observed, overall ability of intramuscular fatty acid mobilization and fatty acid oxidation were enhanced by an 8-week KD feeding, thus contributed to exercise capacity. Compared to a glucose-centered metabolic system, a long-term KD feeding leads to establishment of a fatty acid oxidation-centered metabolic system. Metabolic flexibility is used as a term for the ability to adapt to conditional change in metabolic demand, and an 8-week KD helped established lipid-focused metabolic system through keto-adaption, thus increasing the metabolic flexibility. This is not a denial for the conception “glycogen loading” before competitions, while adequate KD meal may help our body to be more flexible during fuel choosing. Moderate training may enhance the ability to utilize ketone bodies as well as fatty acid, or to increase fatty acid mobilization from adipose tissue. Further investigation is urged to be carried out.   Conclusions In the present study, we investigated how an 8-week KD remodeled adipose and muscle metabolic adaptation towards ketolysis, lipolysis and fatty acid oxidation under the circumstance of exhaustive exercise. Along with enhanced fatty acid oxidation capacity, KD also enhanced fatty acid mobilization capacity, ketolysis and lipolysis. These results revealed that an 8-week KD administration enhance exercise performance by up-regulated ketolytic and free fatty acid oxidation ability, indicating KD being a promising diet approach in athletes

PO-114 Effects of one-time exhaustive exercise on peripheral drive in rats

Objective In this study, we observed the effects of one-time exhaustive exercise on the grip strength, the time of grabbing, and the changes of the electromyography (EMG) of the hind limb flexor muscles and the flexor elbow muscles of the forelimbs, and revealed the effects of exercise fatigue on the peripheral motor drive level. Methods Male SD rats finished exhaustive fatigue exercise. A one-time exhaustive treadmill exercise fatigue model was established after one-week adaptive training in rats. The model was established by the modified Bedford incremental load motion program of the laboratory. The load is divided into 3 levels: the first stage movement speed 8.2 m/min, exercise time 15 min; second stage speed 15 m/min, exercise time 15 min; third stage speed at 20 m/min, exercise to exhaustion. At the same time, a miniature wireless acceleration sensor (18g) was worn in the tail of the rat to monitor the acceleration change of the running direction of the rat while running on the running platform. Three consecutive parallel experiments were performed using a rat grip tester (BioSEB GS3) to measure and compare the maximal muscle strength changes of the limbs before and after exercise fatigue in rats. Compared the static contraction of the rat muscle before and after exercise fatigue to overcome the length of time and gravity of the rod, and evaluated the muscle endurance after training the rats to learn to grab the rod. The EMG,square root amplitude (maxRMS), frequency domain analysis of EMG median frequency (MDF) and mean frequency (MPF) of the hind limb flexor and the forelimb flexor muscles (EMG) was measured by the Italian BTS FREEEMG ultra-miniature wireless surface electromyography tester to predict peripheral muscle tone and drive level. Results 1) The maximum holding force of the rat in resting state was 68.53 N/Kg, and the gripping force was significantly decreased (p<0.05) and reduced to 25.47 N/Kg after exercise fatigue .2) Exercise fatigue has a significant effect on the static grab time of rats. The rat has a grab time of 287.65s in a quiet state, and can only last for 27.78s after fatigue, and even can hardly maintain static contraction. The maxRMS of hindlimb flexor muscles in rats was significantly lower than that before fatigue (P<0.05) at rest, and there was no difference in forelimb flexor elbow muscle groups. MDF and MDF of forelimb flexor elbow muscle group and hind limb flexor muscle group were significant increased (P<0.05). 4) MaxRMS MDF and MDF of hind limb flexor muscle group and forelimb flexor elbow muscle group were significantly lower than those before fatigue (P<0.05) under the state of grabbing rod. Conclusions The sprinting ability in the running direction,maximum gripping force and grabbing time of the rats decreased significantly after exercise fatigue, revealing that the fatigue of the muscles may cause the decrease of the muscle static contraction ability. The inability of the hind limbs to maintain standing with exercise fatigue may be related to a significant decrease in hindlimb tension, and it was found that there was an explosive discharge and the phenomenon of tonic contraction in the muscles at rest. The muscle endurance and tension of the muscles were significantly reduced, and the contraction frequency of the muscle movement unit decreased significantly after exercise fatigue, causing insufficient peripheral driving level . (NSFC31401018, SKXJX2014014, [email protected])

Direct Conversion of Mouse Astrocytes Into Neural Progenitor Cells and Specific Lineages of Neurons

Background: Cell replacement therapy has been envisioned as a promising treatment for neurodegenerative diseases. Due to the ethical concerns of ESCs-derived neural progenitor cells (NPCs) and tumorigenic potential of iPSCs, reprogramming of somatic cells directly into multipotent NPCs has emerged as a preferred approach for cell transplantation. Methods: Mouse astrocytes were reprogrammed into NPCs by the overexpression of transcription factors (TFs) Foxg1, Sox2, and Brn2. The generation of subtypes of neurons was directed by the force expression of cell-type specific TFs Lhx8 or Foxa2/Lmx1a. Results: Astrocyte-derived induced NPCs (AiNPCs) share high similarities, including the expression of NPC-specific genes, DNA methylation patterns, the ability to proliferate and differentiate, with the wild type NPCs. The AiNPCs are committed to the forebrain identity and predominantly differentiated into glutamatergic and GABAergic neuronal subtypes. Interestingly, additional overexpression of TFs Lhx8 and Foxa2/Lmx1a in AiNPCs promoted cholinergic and dopaminergic neuronal differentiation, respectively. Conclusions: Our studies suggest that astrocytes can be converted into AiNPCs and lineage-committed AiNPCs can acquire differentiation potential of other lineages through forced expression of specific TFs. Understanding the impact of the TF sets on the reprogramming and differentiation into specific lineages of neurons will provide valuable strategies for astrocyte-based cell therapy in neurodegenerative diseases

Reduced Energy Metabolism Impairs T Cell-Dependent B Cell Responses in Patients With Advanced HBV-Related Cirrhosis

Background and AimsPatients with decompensated HBV-related liver cirrhosis (HBV D-LC) showed compromised immune responses, which manifested as a proneness to develop infections and hyporesponsiveness to vaccines, resulting in accelerated disease progression. The alterations in T cell-dependent B cell responses in this pathophysiological process were not well understood. This study aimed to investigate T cell-dependent B cell responses in this process and discuss the mechanism from the perspective of metabolism.MethodsChanges in phenotypes and subsets of peripheral B cells between HBV D-LC patients and healthy controls (HCs) were compared by flow cytometry. Isolated B cells were activated by coculture with circulating T follicular (cTfh) cells. After coculture, the frequencies of plasmablasts and plasma cells and immunoglobin levels were analyzed. Oxidative phosphorylation (OXPHOS) and glycolysis were analyzed by a Seahorse analyzer. Mitochondrial function and the AKT/mTOR pathway were analyzed by flow cytometry.ResultsThe proliferation and differentiation capacities of B cells after T cell stimulation were impaired in D-LC. Furthermore, we found that B cells from D-LC patients showed reductions in OXPHOS and glycolysis after activation, which may result from reduced glucose uptake, mitochondrial dysfunction and attenuated activation of the AKT/mTOR pathway.ConclusionsB cells from HBV D-LC patients showed dysfunctional energy metabolism after T cell-dependent activation. Understanding the regulations of B cell metabolic pathway and their changes may provide a new direction to rescue B cell hyporesponsiveness in patients with HBV D-LC, preventing these patients be infected and improving sensitivity to vaccines

The Epigenetic Modifier PRDM5 Functions as a Tumor Suppressor through Modulating WNT/β-Catenin Signaling and Is Frequently Silenced in Multiple Tumors

BACKGROUND: PRDM (PRDI-BF1 and RIZ domain containing) proteins are zinc finger proteins involved in multiple cellular regulations by acting as epigenetic modifiers. We studied a recently identified PRDM member PRDM5 for its epigenetic abnormality and tumor suppressive functions in multiple tumorigeneses. METHODOLOGY/PRINCIPAL FINDINGS: Semi-quantitative RT-PCR showed that PRDM5 was broadly expressed in human normal tissues, but frequently silenced or downregulated in multiple carcinoma cell lines due to promoter CpG methylation, including 80% (4/5) nasopharyngeal, 44% (8/18) esophageal, 76% (13/17) gastric, 50% (2/4) cervical, and 25% (3/12) hepatocellular carcinoma cell lines, but not in any immortalized normal epithelial cell lines. PRDM5 expression could be restored by 5-aza-2'-deoxycytidine demethylation treatment in silenced cell lines. PRDM5 methylation was frequently detected by methylation-specific PCR (MSP) in multiple primary tumors, including 93% (43/46) nasopharyngeal, 58% (25/43) esophageal, 88% (37/42) gastric and 63% (29/46) hepatocellular tumors. PRDM5 was further found a stress-responsive gene, but its response was impaired when the promoter was methylated. Ectopic PRDM5 expression significantly inhibited tumor cell clonogenicity, accompanied by the inhibition of TCF/β-catenin-dependent transcription and downregulation of CDK4, TWIST1 and MDM2 oncogenes, while knocking down of PRDM5 expression lead to increased cell proliferation. ChIP assay showed that PRDM5 bound to its target gene promoters and suppressed their transcription. An inverse correlation between the expression of PRDM5 and activated β-catenin was also observed in cell lines. CONCLUSIONS/SIGNIFICANCE: PRDM5 functions as a tumor suppressor at least partially through antagonizing aberrant WNT/β-catenin signaling and oncogene expression. Frequent epigenetic silencing of PRDM5 is involved in multiple tumorigeneses, which could serve as a tumor biomarker

MAGE-A cancer/testis antigens inhibit MDM2 ubiquitylation function and promote increased levels of MDM4

Melanoma antigen A (MAGE-A) proteins comprise a structurally and biochemically similar sub-family of Cancer/Testis antigens that are expressed in many cancer types and are thought to contribute actively to malignancy. MAGE-A proteins are established regulators of certain cancer-associated transcription factors, including p53, and are activators of several RING finger-dependent ubiquitin E3 ligases. Here, we show that MAGE-A2 associates with MDM2, a ubiquitin E3 ligase that mediates ubiquitylation of more than 20 substrates including mainly p53, MDM2 itself, and MDM4, a potent p53 inhibitor and MDM2 partner that is structurally related to MDM2. We find that MAGE-A2 interacts with MDM2 via the N-terminal p53-binding pocket and the RING finger domain of MDM2 that is required for homo/hetero-dimerization and for E2 ligase interaction. Consistent with these data, we show that MAGE-A2 is a potent inhibitor of the E3 ubiquitin ligase activity of MDM2, yet it does not have any significant effect on p53 turnover mediated by MDM2. Strikingly, however, increased MAGE-A2 expression leads to reduced ubiquitylation and increased levels of MDM4. Similarly, silencing of endogenous MAGE-A expression diminishes MDM4 levels in a manner that can be rescued by the proteasomal inhibitor, bortezomid, and permits increased MDM2/MDM4 association. These data suggest that MAGE-A proteins can: (i) uncouple the ubiquitin ligase and degradation functions of MDM2; (ii) act as potent inhibitors of E3 ligase function; and (iii) regulate the turnover of MDM4. We also find an association between the presence of MAGE-A and increased MDM4 levels in primary breast cancer, suggesting that MAGE-A-dependent control of MDM4 levels has relevance to cancer clinically

Exploring the pathogenesis of colorectal carcinoma complicated with hepatocellular carcinoma via microarray data analysis

Background: Despite the increasing number of research endeavors dedicated to investigating the relationship between colorectal carcinoma (CRC) and hepatocellular carcinoma (HCC), the underlying pathogenic mechanism remains largely elusive. The aim of this study is to shed light on the molecular mechanism involved in the development of this comorbidity.Methods: The gene expression profiles of CRC (GSE90627) and HCC (GSE45267) were downloaded from the Gene Expression Omnibus (GEO) database. After identifying the common differentially expressed genes (DEGs) of psoriasis and atherosclerosis, three kinds of analyses were performed, namely, functional annotation, protein‐protein interaction (PPI) network and module construction, and hub gene identification, survival analysis and co-expression analysis.Results: A total of 150 common downregulated differentially expressed genes and 148 upregulated differentially expressed genes were selected for subsequent analyses. The significance of chemokines and cytokines in the pathogenesis of these two ailments is underscored by functional analysis. Seven gene modules that were closely connected were identified. Moreover, the lipopolysaccharide-mediated signaling pathway is intricately linked to the development of both diseases. Finally, 10 important hub genes were identified using cytoHubba, including CDK1, KIF11, CDC20, CCNA2, TOP2A, CCNB1, NUSAP1, BUB1B, ASPM, and MAD2L1.Conclusion: Our study reveals the common pathogenesis of colorectal carcinoma and hepatocellular carcinoma. These common pathways and hub genes may provide new ideas for further mechanism research