Efectos de la proteína whey, leucina y entrenamiento de fuerza en la composición corporal / Effects of whey protein, leucine and resistance training on body composition

INTRODUCCIÓN: Los deportistas y las personas activas que entrenan la fuerza para lograr un aumento de su masa muscular tienen requerimientos calóricos y proteicos aumentados, por esta razón emplean diversas estrategias como la suplementación con proteína whey aislada y aminoácidos aislados como la leucina después del entrenamiento sustentados en la premisa de que promueven el incremento en la síntesis de proteínas musculares, y por ende un aumento de la masa magra. OBJETIVO: Comparar cambios de la composición corporal aplicando el método de Absorciometría Dual de Rayos X (DEXA) en personas físicamente activas expuestas a un programa de entrenamiento de la fuerza con la suplementación de proteína whey aislada y/o leucina durante 12 semanas. METODOLOGÍA: estudio aleatorizado prospectivo longitudinal en un grupo de 10 varones físicamente activos, los cuales fueron asignados al azar en 3 grupos de suplementación con proteína whey aislada, leucina o la mezcla de ambas bajo el mismo protocolo de entrenamiento de la fuerza. Se evaluó la composición corporal en 3 momentos: semana 0, semana 6 y semana 12 de la intervención. RESULTADOS: En las condiciones evaluadas no se observaron cambios estadísticamente significativos de las variables de composición corporal y fuerza en los diferentes protocolos de suplementación: leucina, proteína whey aislada con leucina y proteína whey aislada. CONCLUSIONES: Se observaron resultados superiores de la masa magra y la fuerza en el grupo de la suplementación de proteína whey aislada con leucina con respecto a los demás grupos, cabe aclarar que por el tamaño de muestra no alcanzó a ser estadísticamente significativo.

INTRODUCTION: Athletes and active people who work hard to increase their muscle mass have increased caloric and protein requirements. For this reason, they use different strategies such as whey protein isolate supplementation and/or branched-chain amino acids after sustained training on the premise that they promote an increase in muscle protein synthesis, and therefore an increase in lean mass. OBJECTIVE: To compare body composition changes in physically active people exposed to a strength training program withwhey protein isolate and/or leucine supplementation for 12 weeks. METHODS: Longitudinal prospective randomized study in a group of 10 physically active males, which were randomly assigned to 3 supplementation groups with whey protein isolate, leucine, or a mixture of both, under the same strength training protocol. Body composition was evaluated in 3 moments: week 0, week 6 and week 12 of the intervention. RESULTS: In the evaluated conditions, no statistically significant changes were observed in the variables of body composition and strength based on the different supplementation protocols: Leucine, protein with leucine and protein. CONCLUSIONS: Compared with the other groups, higher results were observed in lean mass and strength in group using the whey protein isolate supplementation with leucine. It should be noted that due to the size of the sample, the difference was not statistically significant.

Correlation of serum antibody titers with clinical features and prognosis of anti leucine-rich glioma inactivated 1 antibody encephalitis / 中华神经科杂志

Objective:To analyze the relationship between serum antibody titers, clinical characteristics, and prognosis in patients with encephalitis induced by anti leucine-rich glioma inactivated 1 (LGI1) antibody.Methods:Clinical data of 20 patients diagnosed with encephalitis in the Department of Neurology, First Hospital of Shanxi Medical University from February 2015 to February 2021 were collected and retrospectively analyzed. Patients were divided into 2 groups based on their serum anti LGI1 antibody titers, namely the high titer group (1∶100, 1∶320) and the low titer group (1∶10, 1∶32). The clinical characteristics, laboratory test results, and prognosis of the 2 groups of patients were compared. Relusts The age of the 20 patients with anti LGI1 antibody encephalitis ranged from 27 to 69 (53.5±11.2) years, with a male to female ratio of 1∶1. There were 9 patients in the low titer group and 11 patients in the high titer group. There was no statistically significant difference in the types and quantities of clinical symptoms between the 2 groups. Patients in the high titer group were more prone to abnormal lesions on cranial magnetic resonance imaging (10/11 vs 3/9, P=0.014). There was no statistically significant difference between the 2 groups in the presence or absence of cerebrospinal fluid anti LGI1 antibodies (9/11 vs 4/9, P=0.160). During the follow-up, it was found that 1/20 patient died of pulmonary embolism, 7/20 of patients were able to recover to their predisease state, and 9/20 of patients had residual memory impairment. In the high titer group, 3/11 of patients experienced recurrence, while there was no recurrence in the low titer group. There was no statistically significant difference in the neurological prognosis between the 2 groups at 3 months of discharge and follow-up (the number of patients whose modified Rankin Scale score≤2: 10/10 vs 8/9, P=0.474). Conclusions:Patients with high serum anti LGI1 antibody titers are more likely to develop intracerebral lesions. Higher antibody titers may be associated with a higher risk of disease recurrence. There was no significant correlation between serum antibody titers and neurological outcomes.

Causal relationship between branched-chain amino acids and peripheral atherosclerosis： a Mendelian randomization study / 上海预防医学

ObjectiveWe conducted a two-sample Mendelian randomization （MR） study to assess the causal relationship between circulating isoleucine， leucine and valine levels and the risk of peripheral atherosclerosis. MethodsBased on the large-scale genome-wide association study （GWAS） database， single nucleotide polymorphisms （SNPs） closely related to the circulating levels of isoleucine， leucine and valine were identified as instrumental variables （IVs）. Two-sample MR analysis applying the inverse variance weighted （IVW） method and the weighted median estimator （WME） method were performed to estimate the causal relationship between the risk of peripheral atherosclerosis and the exposure with more than three SNPs that were available as IVs. The pleiotropy was evaluated by using the MR-Egger regression and MR-PRESSO method， and the leave-one-out method was used in sensitivity analysis. ResultsFour， one and one SNPs were identified as IVs for circulating isoleucine， leucine and valine levels， respectively. For isoleucine， the IVW model demonstrated there was no evidence of heterogeneity among the IVs （P=0.715）， and there was a significant causal relationship between the increase of circulating isoleucine level and a higher risk of peripheral atherosclerosis risk. Per every 1 elevated standard deviation （SD） of circulating isoleucine level resulted in increasing 31% of peripheral atherosclerosis risk （OR=1.31， 95%CI： 1.07‒1.61）. Similarly， the OR（95%CI） was 1.33 （1.04‒1.71） in the WME model. The MR-Egger regression and MR-PRESSO analysis indicated no evidence of pleiotropy in IVs （all P>0.05）. The result of the leave-one-out sensitivity analysis was stable. The Wald ratio model displayed that the causal relationship between circulating leucine and valine levels and the risk of peripheral atherosclerosis was not statistically significant. The OR （95%CI） for leucine and valine was 1.13 （0.78‒1.63） and 1.11 （0.82‒1.50）， respectively. ConclusionThere is a significant causal relationship between the increase of circulating isoleucine level and a higher peripheral atherosclerosis risk. The causal relationships between circulating leucine and valine levels and the risk of peripheral atherosclerosis need to be further confirmed in future studies.

Transcription factor EB attenuates neuronal damage following cerebral ischemia via regulating autophagy-lysosomal pathway / 国际脑血管病杂志

The disorder of autophagy lysosomal pathway (ALP) is an important pathogenesis of neuronal damage after cerebral ischemia, and the restoration of ALP may alleviate neuronal damage after cerebral ischemia. As the main transcription factor regulating ALP, transcription factor EB (TFEB) can directly regulate autophagosome generation, autophagosome-lysosome fusion, and autophagic flux by regulating the expression of autophagic genes and lysosomal genes. Therefore, regulating TFEB can alleviate ALP dysfunction and thereby reduce cerebral ischemic damage. This article reviews the structure, biological function of TFEB and its role in regulating ALP to alleviate neuronal damage after cerebral ischemia.

LRG1 inhibits hepatic macrophage activation by enhancing TGF-β1 signaling to alleviate MAFLD in mice / 南方医科大学学报

OBJECTIVE@#To explore the effect of leucine-rich α-2-glycoprotein (LRG1) derived from hepatocytes on activation of hepatic M1 Kupffer cells.@*METHODS@#A metabolic dysfunction-associated fatty liver disease (MAFLD) model was established in BALB/c mice by high-fat diet (HFD) feeding for 16 weeks. Oleic acid was used to induce steatosis in primary cultures of mouse hepatocytes. The mRNA and protein expressions of LRG1 in mouse liver tissues and hepatocytes were detected by real-time PCR and Western blotting. Primary hepatic macrophages were stimulated with the conditioned medium (CM) from steatotic hepatocyte along with LRG1 or transforming growth factor-β1 (TGF-β1), or both for 24 h, and the expression levels of inducible nitric oxide synthase (iNOS) was detected with Western botting, and the mRNA expressions of iNOS, chemokine ligand 1 (CXCL-1) and interleukin-1β (IL-1β) were measured by RT-PCR. The MAFLD mice were injected with LRG1 (n=6), TGF-β1 (n=6), or both (n=6) through the caudal vein, and the live tissues were collected for HE staining and immumohistochemical detection of F4/80 expression; the mRNA expressions of iNOS, CXCL-1 and IL-1β in liver tissues were detected using RT-PCR.@*RESULTS@#The mRNA and protein expression levels of LRG1 were significantly downregulated in the liver tissues of MAFLD mice and steatotic hepatocytes (P < 0.05). Treatment of the hepatic macrophages with CM from steatosis hepatocytes significantly enhanced the mRNA expression levels of iNOS, CXCL-1 and IL-1β, and these changes were significantly inhibited by the combined treatment with TGF-β1 and LRG1 (P < 0.05). In MAFLD mice, injections with either LRG1 or TGF-β1 alone reduced hepatic lipid deposition and intrahepatic macrophage infiltration, and these effects were significantly enhanced by their combined treatment, which also more strongly inhibited the mRNA expression levels of iNOS, CXCL-1 and IL-1β (P < 0.05).@*CONCLUSION@#LRG1 inhibits hepatic macrophage infiltration by enhancing TGF-β1 signaling to alleviate fatty liver inflammation in MAFLD mice.

Expression of LRG-1 in mice with hypertensive renal damage and its significance / 中南大学学报(医学版)

OBJECTIVES@#Long-term elevated blood pressure may lead to kidney damage, yet the pathogenesis of hypertensive kidney damage is still unclear. This study aims to explore the role and significance of leucine-rich alpha-2-glycoprotein-1 (LRG-1) in hypertensive renal damage through detecting the levels of LRG-1 in the serum and kidney of mice with hypertensive renal damage and its relationship with related indexes.@*METHODS@#C57BL/6 mice were used in this study and randomly divided into a control group, an angiotensin II (Ang II) group, and an Ang II+irbesartan group. The control group was gavaged with physiological saline. The Ang II group was pumped subcutaneously at a rate of 1.5 mg/(kg·d) for 28 days to establish the hypertensive renal damage model in mice, and then gavaged with equivalent physiological saline. The Ang II+irbesartan group used the same method to establish the hypertensive renal damage model, and then was gavaged with irbesartan. Immunohistochemistry and Western blotting were used to detect the expression of LRG-1 and fibrosis-related indicators (collagen I and fibronectin) in renal tissues. ELISA was used to evaluate the level of serum LRG-1 and inflammatory cytokines in mice. The urinary protein-creatinine ratio and renal function were determined, and correlation analysis was conducted.@*RESULTS@#Compared with the control group, the levels of serum LRG-1, the expression of LRG-1 protein, collagen I, and fibronectin in kidney in the Ang II group were increased (all P<0.01). After treating with irbesartan, renal damage of hypertensive mice was alleviated, while the levels of LRG-1 in serum and kidney were decreased, and the expression of collagen I and fibronectin was down-regulated (all P<0.01). Correlation analysis showed that the level of serum LRG-1 was positively correlated with urinary protein-creatinine ratio, blood urea nitrogen, and blood creatinine level in hypertensive kidney damage mice. Serum level of LRG-1 was also positively correlated with serum inflammatory factors including TNF-α, IL-1β, and IL-6.@*CONCLUSIONS@#Hypertensive renal damage mice display elevated expression of LRG-1 in serum and kidney, and irbesartan can reduce the expression of LRG-1 while alleviating renal damage. The level of serum LRG-1 is positively correlated with the degree of hypertensive renal damage, suggesting that it may participate in the occurrence and development of hypertensive renal damage.

Effects of single ingestion of arginine on mTORC1 activation in rat fast- and slow-twitch muscles / 体力科学

This study aimed to examine whether a single ingestion of arginine activates the mammalian target of rapamycin complex 1 (mTORC1) in rat fast- and slow-twitch skeletal muscles. In the first experiment, the rats were orally administered arginine (3 or 10 mmol/kg body weight) in water. The plantaris, gastrocnemius, and soleus muscles were excised 1 h after the administration. Immunoblot analysis showed that the administration with a higher dose (10 mmol/kg) resulted in increased phosphorylation of ribosomal S6 kinase (S6K) and ribosomal protein S6 only in the soleus muscles. The amounts of cellular arginine sensor for mTORC1 subunit 1 (CASTOR1) expressed were similar in these three muscles. In the second experiment, the plantaris and soleus muscles were excised 1 h after the administration of 10 mmol/kg of arginine. The binding of CASTOR1 to the GATOR2 complex was not detected in either muscle in co-immunoprecipitation and immunoblot analyses, irrespective of arginine administration. In the third experiment, a role of nitric oxide (NO) was elucidated. Treatment with an inhibitor of NO synthase blocked the arginine-induced increase in S6K phosphorylation. These results indicate that a single ingestion of arginine is capable of activating mTORC1 only in slow-twitch muscles and suggest that the activation may be mediated via NO, but not via the CASTOR1-GATOR2 complex pathway.

Expression of BZW2 and IVD in hepatocellular carcinoma tissues and effect on prognosis of liver transplant recipients with hepatocellular carcinoma / 器官移植

Objective To investigate the expression levels of basic leucine zipper and W2 domain 2 (BZW2) and isovaleryl-CoA dehydrogenase (IVD) in hepatocellular carcinoma (HCC) and evaluate their effect on clinical prognosis of liver transplant recipients with HCC. Methods Pathological specimens and clinical data of 87 liver transplant recipients with HCC were collected and retrospectively analyzed. The recurrence and metastasis of HCC after liver transplantation were assessed. Immunohistochemical staining was used to detect the expression levels of BZW2 and IVD. The relationship between BZW2, IVD and clinicopathological parameters of HCC and their effect on postoperative recurrence and clinical prognosis of the recipients was analyzed. Results Among 87 recipients, 31 cases recurred with a recurrence rate of 36%. HCC recurred at postoperative 2-49 months and the median recurrence time was postoperative 7 months. Immunohistochemical staining demonstrated that the positive expression rate of BZW2 in the HCC tissues was significantly higher than that in normal liver tissues (76% vs. 30%), and the positive expression rate of IVD was significantly lower compared with that in normal liver tissues (51% vs. 69%) (both P < 0.01). BZW2 expression was significantly correlated with tumor diameter and tumor capsule (both P < 0.05), whereas IVD expression was significantly associated with tumor diameter, alpha-fetoprotein (AFP) level, tumor, node and metastasis (TNM) staging and whether vascular invasion was found or not (all P < 0.05). In the high BZW2 expression group, the cumulative recurrence rate of HCC was significantly higher and the cumulative survival rate was significantly lower than those in the low BZW2 expression group. In the low IVD expression group, the cumulative recurrence rate of HCC was significantly higher and the cumulative survival rate was significantly lower compared with those in the high IVD expression group (all P < 0.05). Conclusions The expression level of BZW2 protein is up-regulated, whereas that of IVD protein is down-regulated in the HCC tissues. Moreover, the cumulative recurrence rate of HCC is relatively high and the cumulative survival rate is relatively low in liver transplant recipients with high BZW2 expression and low IVD expression.

Progress of the possible mechanism of leucine-rich repeat kinase 2 affecting thyroid cancer via regulating autophagy / 肿瘤研究与临床

Thyroid cancer is the most common malignant tumor of the endocrine system, and the incidence is increasing year by year, which seriously threatens people's health. Autophagy is a programmed mode of death that can be used as a potential target for anti-tumor therapy and plays an important regulatory role. Leucine-rich repeat kinase 2 (LRRK2) is a protein kinase encoded by PARK8 gene. The recent studies have confirmed that autophagy is closely related to thyroid cancer. This paper analyzes the possible regulatory mechanism of LRRK2 affecting thyroid cancer through autophagy, providing new ideas for basic research and clinical diagnosis and treatment of thyroid cancer.

A case of limbic encephalitis with positive anti-leucine-rich glioma inactivated 1 protein antibody and anti-myelin oligodendrocyte glycoprotein antibody / 中华神经科杂志

A case of limbic encephalitis with positive anti-leucine-rich glioma inactivated 1 protein (LGI1) antibody and anti-myelin oligodendrocyte glycoprotein (MOG) antibody was reported. The patient was a middle-aged male with a history of retinal vein occlusion. The main symptoms were temporal lobe epilepsy, facial arm dystonia, autonomic nerve dysfunction. Magnetic resonance imaging showed long T 2 signal in the right hippocampus without enhancement and normal perfusion. Electroencephalogram showed paroxysmal slow wave and sharp slow wave in interictal period. Blood anti-MOG antibody, blood and cerebrospinal fluid anti-LGI1 antibody were double positive. The main diagnosis was limbic encephalitis. After treatment with hormone and gamma globulin, the symptoms were improved and double antibodies were turned negative. Anti-LGI1/MOG double positive cases are rare, and the clinical manifestations and imaging manifestations of double positive antibody cases are not completely consistent with those with each single antibody, with different characteristics. This report can help clinicians enhance awareness.

Morvan syndrome with positive anti LGI1/CASPR2 antibodies in serum/cerebrospinal fluid:a case report and literature review / 中华内科杂志

To report a typical case of Morvan syndrome with positive anti-leucine rich glioma-inactivated 1(LGI1) and contactin-associated protein 2 (CASPR2) antibodies in serum and cerebrospinal fluid. A 39-years-old female initially presented weakness of extremeties. The main symptoms included paroxysmal limb pain, wheezing, itching, muscle twitching, epilepsy, hypomnesia, dysphoria, apathy, intractable insomnia, salivation and sweating. Tests of electrolytes found hypokalemia (2.7-3.1 mmol/L) and hyponatremia (130-136 mmol/L). Arterial blood gas analysis showed hypoxemia (oxygen saturation 50%-70%). Total thyroxine (TT4) was elevated to 207 nmol/L with positive thyroid peroxidase antibody (TPO-Ab) and thyroglobulin antibody (TG-Ab). LGI1and CASPR2 antibodies (CBA method) were positive in both serum and cerebrospinal fluid, and the remaining antibodies related to autoimmune encephalitis and paraneoplastic syndrome were negative. Head MRI was almost normal, while mild abnormalities were found in electroencephalogram. Electromyography showed slightly increased voltage of left quadriceps motor unit potential. After treated with corticosteroids, IVIG and mycophenolate mofetil, the patient completely improved. Cognitive function scores recovered from MoCA/MMSE (16/24) to MoCA/MMSE (26/29). Positivity of LGI1/CASPR2 antibodies both in serum/cerebrospinal fluid are rarely seen in patients with Morvan syndrome. Steroids and immunosuppressants are suggested for treatment as early as possible.

The expression of leucine zipper EF-hand domain transmembrane protein 1, sodium hydrogen exchange protein 1 and adenylate cyclase related protein 2 in gastric cancer tissues and their predictive value for postoperative recurrence / 中国医师进修杂志

Objective:To investigate the expression of leucine zipper EF-hand domain transmembrane protein 1 (LETM1), sodium hydrogen exchange protein 1 (NHE-1) and adenylate cyclase related protein 2 (CAP2) in gastric cancer tissues and the predictive value for postoperative recurrence.Methods:The clinical data of 92 patients with early gastric cancer who underwent surgical treatment from January 2017 to January 2020 in Jiangsu Haimen People′s Hospital were retrospectively analyzed. According to the recurrence condition 6 months after operation, the patients were divided into recurrence group (16 cases) and non recurrence group (76 cases). The expression levels of LETM1, NHE-1 and CAP2 mRNA in cancer tissues and adjacent tissues were detected by real time fluorescent quantitative polymerase chain reaction. Multifactor Logistic regression analysis was used to analyze the related influencing factors of postoperative recurrence. The receiver operating characteristic (ROC) curve was drawn, and the effectiveness of LETM1, NHE-1 and CAP2 mRNA in predicting recurrence was analyzed. The Kaplan-Meier survival curve was drawn, and the survival rate was analyzed in patients with different expression levels of LETM1, NHE-1 and CAP2 mRNA.Results:The LETM1, NHE-1 and CAP2 mRNA in cancer tissues were significantly higher than those in adjacent tissues (0.41±0.12 vs. 0.22±0.07, 0.85±0.27 vs. 0.49±0.15 and 0.31±0.10 vs. 0.19±0.06), and there were statistical differences ( P<0.01). The proportion of N 1 stage in recurrent group was significantly higher than that in non recurrent group: 9/16 vs. 22.37% (17/76), and there was statistical difference ( P<0.05); the LETM1, NHE-1 and CAP2 mRNA of cancer tissues in recurrent group were significantly higher than those in non recurrent group (0.61±0.20 vs. 0.37±0.13, 1.24±0.38 vs. 0.77±0.21 and 0.60±0.19 vs. 0.25±0.10), and there were statistical differences ( P<0.01). Multifactor Logistic regression analysis result showed that, after controlled N stages, the LETM1, NHE-1 and CAP2 mRNA were still independent risk factors for postoperative recurrence in patients with gastric cancer ( OR = 1.52, 3.11 and 1.21; 95% CI 1.26 to 1.82, 2.36 to 4.09 and 1.04 to 1.41; P<0.01). ROC curve analysis result showed that the area under the curve (AUC) of LETM1, NHE-1 and CAP2 mRNA for predicting postoperative recurrence in patients with gastric cancer were 0.768, 0.802 and 0.850, respectively. The AUC of combination the indexes for predicting postoperative recurrence in patients with gastric cancer was 0.965. According to the cut-off value of ROC curve, the patients were divided into LETM1 mRNA high expression (>0.54, 30 cases) and low expression (≤0.54, 62 cases), NHE-1 mRNA high expression (>1.09, 35 cases) and low expression (≤1.09, 57 cases), CAP2 mRNA high expression (>0.49, 28 cases) and low expression (≤0.49, 64 cases). Kaplan-Meier survival curve analysis result showed that the survival rates in patients with high expression of LETM1, NHE-1 and CAP2 mRNA were significantly lower than those in patients with low expression (73.33% vs. 98.39%, 80.00% vs. 96.49% and 78.57% vs. 95.31%), and there were statistical differences ( χ2 = 15.08, 6.95 and 6.75, P<0.01). Conclusions:LETM1, NHE-1 and CAP2 mRNA are related to the recurrence of early gastric cancer after surgery. The detection of the 3 markers is expected to provide a new strategy for the prediction of postoperative recurrence.

Effects of small leucine-rich proteoglycans on corneal transparency / 中华实验眼科杂志

Small leucine-rich proteoglycans (SLRPs) are necessary structural ingredients of the cornea, which are vital for the establishment and maintenance of corneal transparency.SLRPs are mainly located in the corneal stroma and can be divided into class Ⅰ, class Ⅱ, and class Ⅲ.The compensatory and cooperative interactions among SLRPs regulate the formation and assembly of stromal collagen fibrils, thereby maintaining the highly ordered arrangement of collagen fibrils, and establishing corneal transparency.Decorin and lumican are the main functional components of class Ⅰ and class Ⅱ SLRPs, respectively, and changes in their expression or abnormities in the structure of their core proteins affect the natural content and arrangement of other stromal extracellular matrix components, ultimately resulting in abnormal fibril formation, assembly, and arrangement, causing corneal opacity.SLRPs can regulate corneal wound healing and stromal matrix remodeling via binding to fibrotic molecules and their receptors, which provides bases for corneal diseases therapy and study of molecular mechanisms of corneal transparency.The bioactivities and the role of SLRPs in corneal transparency were reviewed in this article.

Expression and promoter methylation status of leucine rich repeat containing 55 in pancreatic carcinoma tissue and the clinical value / 中华胰腺病杂志

Objective:To detect the mRNA expression and methylation status of leucine rich repeat containing 55(LRRC55) gene in pancreatic carcinoma tissues, and discuss the clinical value.Methods:Resected pancreatic ductal adenocarcinoma and normal adjacent specimens from 37 patients admitted in General Surgery of First Affiliated Hospital of Naval Medical University were collected from May 2019 to May 2021. Another two normal pancreas specimens and two blood samples from healthy adults were also collected. All patients′ age, gender, tumor location, tumor size, tumor differentiation, TNM staging, lymphatic metastasis, CEA and CA19-9 level were recorded. Bisulfite treatment of genomic DNA and sequencing analysis was used to study methylation patterns in CpG islands of the promoter for LRRC55 gene in fresh tissues from 2 pancreatic adenocarcinoma and adjacent tissues, 2 normal pancreatic tissues, 2 pancreatic cancer cell lines (PaTu8988 and ASPC1). LRRC55 mRNA in 35 pancreatic adenocarcinoma and adjacent tissues was detected by real-time quantitative PCR and the correlations with clinical parameters were analyzed.Results:CpG islands of LRRC55 in pancreatic adenocarcinoma tissues and pancreatic cancer cell lines was highly methylated and the mean methylation rate was 53% and 71%, respectively; while LRRC55 gene in pancreatic adjacent tissues and normal pancreatic tissues was lowly methylated, and the mean methylation rate was 8% and 11%. The relative expression in the pancreatic adenocarcinoma tissues and the paired adjacent normal tissues was 0.21 (0.02, 1.00 ) and 0.98 (0.33, 3.66 ), respectively; the former was significantly lower than the later and the difference was statistically significant ( P=0.003). Correlation analysis showed that LRRC55 mRNA expression level was related to tumor differentiation and CEA, but not correlated with patients′ age, gender, tumor location and size, CA19-9 level, lymphatic metastasis and TNM staging. Conclusions:Pancreatic cancer tissue and cell lines had abnormal methylation of LRRC55 gene; LRRC55 gene hypermethylation was related with its lower mRNA expression level in pancreatic cancer, which was correlated with the tumor differentiation and CEA level. LRRC55 may be a potential suppressor gene for pancreatic cancer.

Eukaryotic expression and antigen epitope prediction of the LRRC15 protein in excretory secretory antigens of Taenia solium cysticercus / 中国血吸虫病防治杂志

OBJECTIVE@#To conduct eukaryotic expression of the leucine-rich repeat containing 15 (LRRC15), a differentially expressed protein in excretory secretory antigens of Taenia solium cysticercus, and predict its antigen epitope.@*METHODS@#The molecular weight, stability, amino acid sequence composition, isoelectric point and T lymphocyte epitope of the LRRC15 protein were predicted using the bioinformatics online softwares ExPASy-PortParam and Protean. The full-length splicing primers were designed using PCR-based accurate synthesis, and the LRRC15 gene was synthesized. The recombinant pcDNA3.4-LRRC15 plasmid was constructed and transfected into HEK293 cells to express the LRRC15 protein. In addition, the LRRC15 protein was characterized by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting.@*RESULTS@#The recombinant pcDNA3.4-LRRC15 plasmid was successfully constructed, which expressed the target LRRC15 protein with an approximately molecular weight of 70 kDa. Bioinformatics prediction with the ExPASy-PortParam software showed that LRRC15 was a hydrophilic protein, which was consisted of 644 amino acids and had a molecular weight of 69.89 kDa and an isoelectric point of 5.6. The molecular formula of the LRRC15 protein was C3073H4942N846O953S28 and had an instability coefficient is 50.3, indicating that LRRC15 was an instable protein. Bioinformatics prediction with the Protean software showed that the dominant T-cell antigen epitopes were located in 292 to 295, 353 to 361, 521 to 526 and 555 to 564 amino acids of the LRRC15 protein, and the T-cell antigen epitopes with a high hydrophilicity, good flexibility, high surface accessibility and high antigenicity index were found in 122 to 131, 216 to 233, 249 to 254, 333 to 343, 358 to 361, 368 to 372, 384 to 386, 407 to 412, 445 to 450, 469 to 481, 553 to 564, 588 to 594, 607 to 617 and 624 to 639 amino acids. Following transfection of the recombinant pcDNA3.4-LRRC15 plasmid into HEK293 cells, SDS-PAGE and Western blotting identified LRRC15 proteins in cell secretory culture media, cell lysis supernatants and sediments. The LRRC15-His fusion protein was purified from the cell culture medium, and SDS-PAGE identified a remarkable band at approximately 70 kDa, while Western blotting successfully recognized the band of the recombinant LRRC15 protein.@*CONCLUSIONS@#The eukaryotic expression and antigen epitope prediction of the LRRC15 protein in the excretory secretory antigens of T. solium cysticercus have been successfully performed, which provides insights into further understandings of its biological functions.

Pharmacological Treatment of Sarcopenia Tratamento medicamentoso da sarcopenia / Tratamento medicamentoso da sarcopenia Pharmacological Treatment of Sarcopenia

Abstract Sarcopenia has been acquiring a growing importance in the scientific literature and in doctors' offices. As the population ages, it becomes increasingly essential to know, prevent, and treat this clinical condition. The purpose of the present review is to bring up the current evidence on the diagnosis of this pathology, in a practical way, as well as the main current treatment options.

Resumo A sarcopenia vem ganhando cada vez mais importância na literatura científica e nos consultórios médicos. Com o envelhecimento da população, essa condição clínica se torna cada vez mais imprescindível de se conhecer, se prevenir e de se tratar. O objetivo desta revisão é trazer as evidências atuais sobre o diagnóstico dessa patologia, de forma prática, bem como as principais opções atuais de tratamento.

Expression of maternal embryonic leucine zipper kinase (MELK) in lung adenocarcinoma and its correlation with prognosis / 国际生物医学工程杂志

Objective:To detect the expression of maternal embryo leucine zipper kinase (MELK) in lung adenocarcinoma, and to explore the clinical prognosis of MELK and lung adenocarcinoma, andto explore the possibility of MELK as a potential biomarker for lung adenocarcinoma.Methods:The mRNA level of MELK in lung adenocarcinoma and normal tissues adjacent to the cancer was analyzed by bioinformatics methods, and the relationship between its expression and the survival rate of patients with lung adenocarcinoma was analyzed. The clinicopathological data of 70 patients with lung adenocarcinoma who underwent surgical treatment were retrospectively analyzed. Immunohistochemical method was used to detect the expression level of MELK protein in lung adenocarcinoma tissue and normal tissues adjacent to the cancer, and to analyze its relationship with clinicopathological characteristics of patients with lung adenocarcinoma.Results:The results of bioinformatics analysis showed that MELK mRNA was significantly highly expressed in lung adenocarcinoma tissue, and was significantly correlated with the overall survival rate ( P=0.009) and disease-free survival rate ( P=0.039) of patients. Immunohistochemical results showed that the expression of MELK in lung adenocarcinoma tissue was significantly higher than that in normal tissues adjacent to the cancer. The high expression of MELK in lung adenocarcinoma tissue was related to tumor size ( P=0.015) and tumor stage ( P=0.006), but not related to age, gender, smoking, tumor differentiation, and lymph node metastasis (all P>0.05). Conclusions:MELK is highly expressed in lung adenocarcinoma tissues and indicates a poor prognosis, and its expression level is related to the tumor stage of lung adenocarcinoma. MELK may serve as a new prognostic biomarker and potential therapeutic targetfor lung adenocarcinoma.

Expression and Significance of Leucine-rich Repeat-containing G-protein Coupled Receptor 5/6 in Wnt Pathway in Children with Acute Lymphoblastic Leukemia / 中国医学科学院学报

Objective To study the expression and significance of leucine-rich repeat-containing G-protein coupled receptor(LGR)5/6 in childhood acute lymphoblastic leukemia(ALL). Methods A total of 39 children who had ALL and achieved complete remission on day 33 after induction therapy were enrolled.The children before induction therapy were considered as the incipient group,and those who achieved complete remission on day 33 by induction therapy were considered as the remission group.According to the degree of risk,they were assigned into 3 groups:low-risk(

Self-assembly and mineralization of full-length human amelogenin and its functional fragments / 华西口腔医学杂志

OBJECTIVES@#To investigate the dynamic process of the self-assembly behaviors of a full-length human amelogenin (AM) and its functional fragments tyrosine-rich amelogenin peptide (TRAP) and leucine-rich amelogenin peptide(LRAP) @*METHODS@#The full-length human AM and its functional fragments, TRAP and LRAP, were reassembled and purified @*RESULTS@#When pH=8, the full-length human AM and TRAP assembly started spontaneously and formed "nanospheres" after 15 min.The nanospheres formed by TRAP existed independently, with a uniform size but without obvious internal structures. The full-length AM was assembled hierarchically, which formed "nanospheres" and further extended in all directions, formed a chain structure, and then aggregated into a net. The self-assembly behavior of LRAP was not obvious. Proteins mostly existed in the form of monomers without "nanosphere" formation. Only few oligomers were observed. The full-length AM was induced independently for 3 days to form rod-shaped HA crystals. TRAP and LRAP proteins were added, after 3 days the crystal elongation was obvious in the c axis, but the growth in plane A and plane B was poor.@*CONCLUSIONS@#The self-assembly and mineralization behaviors of full-length human AM, TRAP, and LRAP were consistent with the directional growth mechanism of HA crystals

Rational design of the C-terminal Loop region of leucine dehydrogenase and cascade biosynthesis L-2-aminobutyric acid / 生物工程学报

Leucine dehydrogenase (LDH) is the key rate-limiting enzyme in the production of L-2-aminobutyric acid (L-2-ABA). In this study, we modified the C-terminal Loop region of this enzyme to improve the specific enzyme activity and stability for efficient synthesis of L-2-ABA. Using molecular dynamics simulation of LDH, we analyzed the change of root mean square fluctuation (RMSF), rationally designed the Loop region with greatly fluctuated RMSF, and obtained a mutant EsLDHD2 with a specific enzyme activity 23.2% higher than that of the wild type. Since the rate of the threonine deaminase-catalyzed reaction converting L-threonine into 2-ketobutyrate was so fast, the multi-enzyme cascade catalysis system became unbalanced. Therefore, the LDH and the formate dehydrogenase were double copied in a new construct E. coli BL21/pACYCDuet-RM. Compared with E. coli BL21/pACYCDuet-RO, the molar conversion rate of L-2-ABA increased by 74.6%. The whole cell biotransformation conditions were optimized and the optimal pH, temperature and substrate concentration were 7.5, 35 °C and 80 g/L, respectively. Under these conditions, the molar conversion rate was higher than 99%. Finally, 80 g and 40 g L-threonine were consecutively fed into a 1 L reaction mixture under the optimal conversion conditions, producing 97.9 g L-2-ABA. Thus, this strategy provides a green and efficient synthesis of L-2-ABA, and has great industrial application potential.