In situ isolation of nuclei or nuclear proteins from adherent cells: a simple, effective method with less cytoplasmic contamination

BACKGROUND: Isolation of nuclei or nuclear proteins is a prerequisite for western blot, nuclear proteome profiling, and other evaluations of nuclear proteins. Here, we developed a simple method for in situ isolation of nuclei or nuclear proteins by in situ removing the extranuclear part of adherent cells via a classical nonionic detergent triton X-100. RESULTS: First, the feasibility of our method was confirmed by confocal microscopy, atomic force microscopy, scanning electron microscopy, dynamic light scattering, immunofluorescence imaging, and time-lapse dynamic observation. Next, the optimal concentration range (approximately 0.1-1% for ~ 10 min) of triton X-100 and the optimal treatment time (< 30 min) of 0.1-1% Triton X-100 for our method were determined via western blotting of eight extra-/ intra-nuclear proteins. Subsequently, the effectiveness, sensitivity, and cytoplasmic contamination of our method were tested by investigating the levels of phosphorylated p65 (a NF-κB subunit) in the nuclei of endothelial or tumor cells treated with/without lipopolysaccharide (LPS) via western blotting and by comparing with a commercial nuclear protein extraction kit (a classical detergent-based method). The data show that compared with the commercial kit our method obtained a higher yield of total nuclear proteins, a higher pP65 level in both control and LPS groups, and much lower content of GAPDH (as a reference for cytoplasmic contamination) in nuclei. CONCLUSIONS: The in situ isolation of nuclei or nuclear proteins from adherent cells in this study is a simple, effective method with less cytoplasmic contamination. This method/strategy has the potential of improving the quality of downstream evaluations including western blotting and proteomic profiling.

Strength training in the lower limbs of soccer players / Treinamento de força nos membros inferiores dos jogadores de futebol / Entrenamiento de fuerza en las extremidades inferiores de los futbolistas

ABSTRACT Introduction: The isometric weight training method combines the benefits of physical endurance and hypertrophy. This teaching method performs the isometric exercise with the appropriate body weight. Objective: Study the effects of isometric training with weights on soccer players' explosive strength of the lower limbs. Methods: Twenty soccer players were randomly selected and divided equally into experimental and control groups. The real training situation of the athletes was used. The experimental group adopted the isometrics teaching method with weight, while the control group used conventional muscle strength training. The lower limb strength of the two groups of athletes was compared before and after the experiment. Results: The extensor force growth rate in the experimental groups was significantly increased at medium, high and low speeds, while the peak torque did not show significant changes. The soccer players had their muscle strength improved. There was a significant difference between the two groups (P<0.05). Conclusion: Isometric weight training significantly improves mid and fast-extensor muscle strength in young soccer players. Level of evidence II; Therapeutic studies - investigation of treatment outcomes.

RESUMO Introdução: O método de ensino por isometria com peso combina os benefícios de resistência física e hipertrofia. Este método de ensino executa o exercício isométrico com o peso corporal adequado. Objetivo: Estudar os efeitos do treinamento isométrico com pesos sobre a força explosiva dos membros inferiores nos jogadores de futebol. Métodos: Selecionou-se 20 jogadores de futebol por amostragem aleatória, divididos igualmente em grupo experimental e controle. Utilizou-se a situação real de treino dos atletas. O grupo experimental adotou o método de ensino por isometria com peso enquanto o grupo de controle utilizou o treinamento convencional de força muscular. A força dos membros inferiores dos dois grupos de atletas foi comparada antes e após o experimento. Resultado: A taxa de crescimento da força extensora nos grupos experimentais foi significativamente aumentada em velocidades médias, altas e baixas, enquanto o torque de pico não apresentou alterações significativas. Os jogadores de futebol tiveram sua força muscular aprimorada. Houve uma diferença significativa entre os dois grupos (P<0,05). Conclusão: O treinamento isométrico com pesos melhora significativamente a força muscular extensora média e rápida nos jovens jogadores de futebol. Nível de evidência II; Estudos terapêuticos - investigação dos resultados do tratamento.

RESUMEN Introducción: El método de enseñanza por isometría con peso combina los beneficios de la resistencia física y la hipertrofia. Este método de enseñanza realiza el ejercicio isométrico con el peso corporal adecuado. Objetivo: Estudiar los efectos del entrenamiento isométrico con pesas sobre la fuerza explosiva de los miembros inferiores en jugadores de fútbol. Métodos: Se seleccionaron 20 jugadores de fútbol por muestreo aleatorio, divididos por igual en grupo experimental y grupo de control. Se utilizó la situación real de entrenamiento de los atletas. El grupo experimental adoptó el método de enseñanza por isometría con peso, mientras que el grupo de control utilizó el entrenamiento de fuerza muscular convencional. Se comparó la fuerza de las extremidades inferiores de los dos grupos de atletas antes y después del experimento. Resultados: La tasa de crecimiento de la fuerza extensora en los grupos experimentales aumentó significativamente a velocidades medias, altas y bajas, mientras que el par máximo no mostró cambios significativos. Los jugadores de fútbol mejoraron su fuerza muscular. Hubo una diferencia significativa entre los dos grupos (P<0,05). Conclusión: El entrenamiento isométrico con pesas mejora significativamente la fuerza de los músculos extensores medios y rápidos en jóvenes futbolistas. Nivel de evidencia II; Estudios terapéuticos - investigación de los resultados del tratamiento.

A highly sensitive bio-barcode immunoassay for multi-residue detection of organophosphate pesticides based on fluorescence anti-quenching / 药物分析学报

Balancing the risks and benefits of organophosphate pesticides(OPs)on human and environmental health relies partly on their accurate measurement.A highly sensitive fluorescence anti-quenching multi-residue bio-barcode immunoassay was developed to detect OPs(triazophos,parathion,and chlorpyrifos)in apples,turnips,cabbages,and rice.Gold nanoparticles were functionalized with monoclonal antibodies against the tested OPs.DNA oligonucleotides were complementarily hybridized with an RNA fluorescent label for signal amplification.The detection signals were generated by DNA-RNA hybridization and ribonuclease H dissociation of the fluorophore.The resulting fluorescence signal en-ables multiplexed quantification of triazophos,parathion,and chlorpyrifos residues over the concen-tration range of 0.01-25,0.01-50,and 0.1-50 ng/mL with limits of detection of 0.014,0.011,and 0.126 ng/mL,respectively.The mean recovery ranged between 80.3％and 110.8％with relative standard deviations of 7.3％-17.6％,which correlate well with results obtained by liquid chromatography-tandem mass spectrometry(LC-MS/MS).The proposed bio-barcode immunoassay is stable,reproducible and reliable,and is able to detect low residual levels of multi-residue OPs in agricultural products.

IL-27 in combination with IL-15 regulates anti-tumor effect of NK92 cells by phosphorylating the STATs pathway / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effect of IL-27 in combination with IL-15 on the anti-tumor effects of NK92 cells and the possible molecular and signaling mechanisms. Methods: NK92 cells with high IL-15 expression (IL-15-NK92 cells) were cultured in different mass concentrations of IL-27 (0, 10, 20, 30 and 60 ng/ml) for 24 h. The effects of IL-27 on IL-15 secretion, migration and proliferation of IL-15-NK92 cells were detected by ELISA, Transwell and CCK-8 assay, respectively. Flow cytometry was used to detect the expression levels of IL-15-NK92 cell surface receptors NKG2D, NKp30 and NKp46, as well as the secretion levels of perforin and granzyme B. LDH method was used to detect the cytotoxic effect of IL-15-NK92 cells on hematologic tumor cells and solid tumor cells, and WB was used to detect the expressions and phosphorylation level of STATs pathway-related proteins. Results: IL-27 at the concentration of 30 ng/ml promoted IL-15-NK92 cells secreting IL-15 (P<0.01), significantly enhanced the cell migration (P<0.05) but inhibited the proliferation of IL-15-NK92 cells (P<0.05). 30 ng/ml IL-27 could significantly promote the expressions of NKG2D, NKp30 and NKp46 on surface of IL-15-NK 92 cells, as well as elevate the secretion of perforin (all P<0.05), but didn’t affect the secretion of granzyme B (P>0.05); moreover, it also significantly enhanced the cytotoxicity of IL-15-NK92 cells against hematologic malignancies and solid tumor cells (P<0.05 or P<0.01), and up-regulated the phosphorylation levels of STAT1, STAT3 and STAT5 (all P<0.01). Conclusion: IL-27 can enhance the cytotoxicity of IL-15-NK92 cells against hematologic tumor cells and solid tumor cells, which might be related with its upregulation of phosphorylation level of STAT1, STAT3 and STAT5 in JAK-STAT pathway and multiple activating receptors in IL-15-NK92 cells.

Exocyst subunit BcSec3 regulates growth, development and pathogenicity in Botrytis cinerea

Botrytis cinerea is a saprophytic plant pathogenic fungus that can infect a variety of crops and cause gray mold,which leads to huge losses worldwide. The role of exocyst in fungal pathogenicity is being revealed. In thisstudy, homologous recombination technology was used to knock out the exocyst subunit BcSec3 of B. cinerea,and it was found that the BcSec3 subunit plays a crucial role in the growth and pathogenicity of B. cinerea.Compared with the wild-type strain B05.10, the mycelial growth ability of the BcSec3 deletion strain wasreduced by up to 49.8%, the conidia production capacity of the deletion strain was severely lost, and nosclerotia was formed. The polygalacturonase, is one of plant cell wall hydrolases, whose activity in BcSec3deletion strain was significantly reduced. In the tomato leaves infection assay in vitro, the lesion area caused bythe BcSec3 deletion strain was only 20% of the wild type after 5 days of infection. Observation by lightmicroscope showed that the morphology of BcSec3 deletion strain mycelium was significantly changed, themycelium became thinner and deformed, and the polarity growth was not obvious. Further observation withlaser confocal microscopy and transmission electron microscopy was conducted. It was found that comparedwith the wild type, the number of vesicles in BcSec3 deleted cells reduced and localization and distribution ofvesicles changed. In mutant cell, vesicles relatively concentrated in the cytoplasm, while in wild-type cellmainly concentrated inside the cell membrane. These evidences indicate that the exocyst subunit BcSec3 playsan important role in the growth, development and pathogenicity of B. cinerea.

Efficacy of different treatment strategies for congenital aortic valve disease in 85 children from a single center: A retrospective cohort study / 中国胸心血管外科临床杂志

@#Objective    To analyze the early- and middle-term prognosis of various surgical methods in children with congenital aortic valve diseases, to provide reference for surgical methods in children with aortic valve stenosis or regurgitation. Methods    The clinical data of 85 children with various aortic valve diseases treated in the Children’s Hospital of Fudan University from January 2005 to December 2018 were retrospectively analyzed. There were 64 males and 21 females, with an average age of 45 months ranging from 5 days to 15 years. Among them 18 patients underwent balloon aortic valvuloplasty (BAV), 8 surgical aortic valvotomy (SAV), 27 aortic valve autogenous pericardium repair, 16 mechanical arterial valve replacement and 16 Ross operation. They were followed up for 6.25±2.76 years. The re-intervention and survival status after different operations were analyzed. Results    There were 3 deaths and 17 reoperations in 85 children. The 5-year survival rate of the patients with SAV, BAV, aortic valve autogenous pericardium repair, mechanical arterial valve replacement and Ross operation was 87.4%, 88.9%, 100.0%, 100.0% and 100.0%, respectively; there was no statistical difference in the early and middle-term survival rates among various operations (P>0.05). The 5-year free from re-intervention rate of the patients with SAV, BAV, aortic valve autogenous pericardium repair, mechanical arterial valve replacement and Ross operation was 44.4%, 18.4%, 100.0%, 66.9% and 80.5%, respectively; there was a statistical difference in the early and middle-term re-intervention rate among various operations (P<0.05). Conclusion    The operation of congenital aortic stenosis or regurgitation needs to be performed according to the pathological changes of the valvular tissues. For children with severe lesions, SAV is recommended for the first intervention. For congenital aortic stenosis, SAV and BAV are both palliative operations which need further evaluation and re-intervention. Ross operation and mechanical arterial valve replacement have low re-intervention rate, and the middle- and long-term follow-up shows that the effect is accurate. Aortic valve autogenous pericardium repair is expected to become a method to delay or replace Ross operation and valve replacement.

Overexpression of histone deacetylase 11 suppresses basal-like breast cancer cell invasion and metastasis / 南方医科大学学报

OBJECTIVE@#Histone deacetylase 11 (HDAC11) is a class Ⅳ member of histone deacetylase family, and its role in regulating cancer cell invasion and metastasis remains unclear. We aimed to investigate the role of HDAC11 in regulating the biological behaviors of basal-like breast cancer (BLBC) cells.@*METHODS@#We analyzed the expression of HDAC11 based on Gene Expression Omnibus (GEO) and the Cancer Genome Atlas (TCGA). The effects of HDAC11 on the cell invasion and metastasis were examined using Transwell assay and in a mouse model. The interaction between HDAC11 and Twist was detected with immunoprecipitation. We identified 2 as a target gene of Twist using promoter luciferase assay and chromatin immunoprecipitation assay.@*RESULTS@#HDAC11 was lowly expressed in BLBC cells. HDAC11 overexpression suppressed BLBC cell invasion and their metastasis in nude mice. Mechanistically, HDAC11 directly interacted with Twist protein, antagonized its pro-invasive function and repressed Twist-induced 2 gene transcription.@*CONCLUSIONS@#Our data suggest that HDAC11 acts as a negative modulator of invasion and metastasis of BLBC cells.

Influence of extracurricular physical exercise on psychological capital, psychological health and social adaptability of college students / 中国学校卫生

Objective@#To investigate the influence of extracurricular physical exercise on psychological capital, psychological health and social adaptability of college students, and to provie the reference for promoting the development of their mental and physical health.@*Methods@#From March 2015 to March 2018, 300 students from Physical Education Institute of Baise University were selected and randomly divided into group A, group B and group C, with 100 students in each group. Group A received exercise prescription training, group B received contract exercise, group C didn’t receive exercise intervention. The scores of Self-reporting Inventory (SCL-90), Psy Cap Questionnaire (PCQ) and College Student Adaptability Inventory (CSAI) were compared among the three groups before intervention and 12 weeks after intervention.@*Results@#After intervention, total average scores of SCL-90 scale in group A and group B were (1.42±0.36) points and (1.40±0.31) points, significantly lower than (1.53±0.34) points in group C (P<0.05), and the score in dimension of somatization, interpersonal sensitivity, anxiety, depression, terror, psychosis among group A and group B was significantly lower than that in group (P<0.05). Group B students had lower score in interpersonal relationship, anxiety and depression than that of group A students (P<0.05). After intervention, total average scores of PCQ scale in group A and group B were (3.11±0.56) points and (3.13±0.54) points, significantly higher than (2.10±0.47) points in group C (P<0.05). No significant differences were found between group A and group B (P>0.05). After intervention, average scores of CSAI in group A and group B were (4.02±0.69) points and (4.09±0.71) points, which were significantly higher than (2.30±0.62) points in group C (P<0.05). No significant differences were observed between group A and group B (P>0.05).@*Conclusion@#Sustained extracurricular physical exercise can significantly improve psychological capital and psychological health among college students, as well as developing social adaptability.

Analysis of parenchymal-sparing hepatectomy in surgical treatment of colorectal liver metastases / 中国肿瘤临床

Objective:To analyze the role of parenchymal-sparing hepatectomy (PSH) in surgical treatment of colorectal liver metasta-ses (CRLM). Methods:All CRLM patients registered in our database who underwent liver resection were assigned into two groups:the PSH group and the major hepatectomy (MH) group. Clinical characteristics were retrospectively analyzed. Surgical outcomes, survival time, and recurrence were compared between the two groups. Results:A total of 377 patients were enrolled. Except for tumor num-bers, no significant difference was found in patient demographics and tumor characteristics between the 2 groups. The rate of PSH in-creased over the study period. Short-term surgical outcomes were better in the PSH group than in the MH group. Surgery time was sig-nificantly shorter (177.5 min vs. 220 min, P=0.000), blood loss was significantly reduced (150 mL vs. 300 mL, P=0.000), and morbidity was significantly reduced (47.4%vs. 64.8%, P=0.008). No significant difference was found in overall survival (OS) time (43 m vs. 41 m, P=0.750) between the 2 groups. Hepatic recurrence-free survival time (21 m vs. 13 m, P=0.344) was similar between the 2 groups. However, local treatment was more frequently performed in the PSH group (42.8%vs. 25.6%, P=0.040) when disease recurred. The 5-year OS was significantly better in patients undergoing local treatment than in those without undergoing local treatment (58 m vs. 24 m, P=0.000), for hepatic recurrence. Conclusion:PSH did not increase hepatic recurrence but improve salvage ability in case of recur-rence with a better short-term surgical outcome. PSH should be the recommended approach at initial hepatectomy.

Uniportal and triportal video-assisted thoracoscopic lobectomy for treatment of non-small cell lung cancer: a comparative study / 中国内镜杂志

Objective To compare the clinical effects of uniportal and triportal video-assisted thoracoscopic lobectomy in treatment of non-small cell lung cancer. Methods From January 2013 to November 2016, 141 non-small cell lung cancer patients underwent uniportal video-assisted thoracoscopic lobectomy (52 cases, uniportal group), and 89 lung cancer patients underwent triportal video-assisted thoracoscopic lobectomy (triportal group). All the cases were performed by the same surgeon. The operative time, intraoperative blood loss, lymph node dissection number, chest drainage duration, postoperative total drainage volume, hospital stay and postoperative complications were recorded and compared between the two groups. Results There were no statistical differences in lymph node dissection number, chest drainage duration, postoperative total drainage volume and postoperative complications after surgery between the uniportal group and the triportal group (P > 0.05). The mean operative time for the uniportal group and the triportal group was (196.1 ± 19.6) minutes and (162.7 ± 18.9) minutes, respectively, P = 0.000. The mean intraoperative blood loss for the uniportal group and the triportal group was (100.3 ± 13.6) ml and (176.5 ± 15.9) ml, respectively, P = 0.000. The hospital stay for the uniportal group and the triportal group was (7.5 ± 1.7) days and (9.2 ± 1.3) days, P = 0.000. Conclusion Uniportal video-assisted thoracoscopic lobectomy can achieve the same effect as triportal video-assisted thoracoscopic lobectomy, though it increased the operation time, however, it has the advantage of less surgical damage to chest wall muscles, intercostal nerves or blood vessels, shorten the postoperative hospital stay, and can be used as a safe and reliable radical surgery for non-small cell lung cancer.

Uniportal and triportal video-assisted thoracoscopic lobectomy for treatment of non-small cell lung cancer: a comparative study / 中国内镜杂志

Objective To compare the clinical effects of uniportal and triportal video-assisted thoracoscopic lobectomy in treatment of non-small cell lung cancer. Methods From January 2013 to November 2016, 141 non-small cell lung cancer patients underwent uniportal video-assisted thoracoscopic lobectomy (52 cases, uniportal group), and 89 lung cancer patients underwent triportal video-assisted thoracoscopic lobectomy (triportal group). All the cases were performed by the same surgeon. The operative time, intraoperative blood loss, lymph node dissection number, chest drainage duration, postoperative total drainage volume, hospital stay and postoperative complications were recorded and compared between the two groups. Results There were no statistical differences in lymph node dissection number, chest drainage duration, postoperative total drainage volume and postoperative complications after surgery between the uniportal group and the triportal group (P > 0.05). The mean operative time for the uniportal group and the triportal group was (196.1 ± 19.6) minutes and (162.7 ± 18.9) minutes, respectively, P = 0.000. The mean intraoperative blood loss for the uniportal group and the triportal group was (100.3 ± 13.6) ml and (176.5 ± 15.9) ml, respectively, P = 0.000. The hospital stay for the uniportal group and the triportal group was (7.5 ± 1.7) days and (9.2 ± 1.3) days, P = 0.000. Conclusion Uniportal video-assisted thoracoscopic lobectomy can achieve the same effect as triportal video-assisted thoracoscopic lobectomy, though it increased the operation time, however, it has the advantage of less surgical damage to chest wall muscles, intercostal nerves or blood vessels, shorten the postoperative hospital stay, and can be used as a safe and reliable radical surgery for non-small cell lung cancer.

Flow-through deep inferior epigastric perforator flaps transplantation for reconstruction of large defects at the extremities / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the clinical application of flow-through deep inferior epigastric perforator flaps for reconstruction of large defects at the extremities.</p><p><b>METHODS</b>The deep inferior and superior epigastric arteries were designed as the axial vessel and the arterial supply to the flap was the paraumbilical perforator artery. Free deep inferior epigastric perforator flaps were harvested in flow- through manners to reconstruct associated arterial defect in the wound. The sensation assessment,Enneking score,and questionnaire of the flap aesthetic were all performed during follow-up period.</p><p><b>RESULTS</b>From December 2011 to September 2012, 5 patients with large defects at extremities were treated. The deep inferior and superior epigastric arteries were designed as the axial vessel and the arterial supply to the flap was the paraumbilical perforator artery. The wound defects ranged form 11 cm x 5 cm to 30 cm x 11 cm. And the flap size ranged from 13 cm x7 cm to 33 cm x 13 cm. All flaps survived completely. The recipient arteries were all bypassed well documented by color Doppler examinations. All cases had 12-24 months' follow-up period. The flaps had good appearance and high aesthetic satisfactory(100%). 12 months after operations, sensation assessment were all S3+, and the Enneking score ratios were 82%-95% ,with 87.2% in average.</p><p><b>CONCLUSIONS</b>Flow-through deep inferior epigastric perforator flaps are reliable and effective for reconstruction of large defects at the extremities with maintenance of the vascular status of the extremities. The flaps can also be designed in transverse or oblique mode for clinical application.</p>

Progression of 14-3-3σ in nasopharyngeal carcinoma / 国际肿瘤学杂志

14-3-3σ,an vital tumor suppressor which is regulated by p53,plays a key role in cell cycle regulation, apoptosis, migration and proliferation, affecting tumor formation, invasion and metastasis. The methylation inactivation of 14-3-3σ is widely recognized as one of the mechanisms of tumorigenesis,and be associated with the metastasis of NPC.

Medical Service Utilization and Trends among Korean Elderly in the Last One Year of Life / 예방의학회지

OBJECTIVES: To analyze medical service utilization and trends among the elderly in the last year of life. METHOD: The subjects of this study were people that had died at the age sixty-five and above between January 1st and June 30th 2000 The names of the deceased and their dates of death were collected from the data of the funeral-expenses-receivers of the National Health Insurance Corporation (NHIC). This data was merged with that of the individual medical expenses of the NHIC. RESULTS: In the first half of 2000, 84.2% of the funeral-expenses-receivers (53, 063) utilized medical services during the year prior to their death; 51.0% (27, 042) were female and 49.0% (26, 021) male. In the last twelve months of life, the medical fees, the number of days receiving medical services and the number of days receiving medicine were 3, 107, 935 Won, 47.88 and 153.21, respectively, for each person. As the age of the groups increased, the level of medical service utilization decreased; the change was more obvious in female group. The level of medical service utilization during the twelve months prior to death drastically increased around the time of death. CONCLUSIONS: This study, from an analysis of the level of medical service utilization prior to death, shows a concentrated volume of medical services during a certain time period prior to death.