Pre-existing interleukin 10 in cerebral arteries attenuates subsequent brain injury caused by ischemia/reperfusion.

Recurrent stroke is difficult to treat and life threatening. Transfer of anti-inflammatory gene is a potential gene therapy strategy for ischemic stroke. Using recombinant adeno-associated viral vector 1 (rAAV1)-mediated interleukin 10 (IL-10), we investigated whether transfer of beneficial gene into the rat cerebral vessels during interventional treatment for initial stroke could attenuate brain injury caused by recurrent stroke. Male Wistar rats were administered rAAV1-IL-10, rAAV1-YFP, or saline into the left cerebral artery. Three weeks after gene transfer, rats were subjected to occlusion of the left middle cerebral artery (MCAO) for 45 min followed by reperfusion for 24 h. IL-10 levels in serum were significantly elevated 3 weeks after rAAV1-IL-10 injection, and virus in the cerebral vessels was confirmed by in situ hybridization. Pre-existing IL-10 but not YFP decreased the neurological dysfunction scores, brain infarction volume, and the number of injured neuronal cells. AAV1-IL-10 transduction increased heme oxygenase (HO-1) mRNA and protein levels in the infarct boundary zone of the brain. Thus, transduction of the IL-10 gene in the cerebral artery prior to ischemia attenuates brain injury caused by ischemia/reperfusion in rats. This preventive approach for recurrent stroke can be achieved during interventional treatment for initial stroke.

Can Yoga Pranayama Practices Improve Burnout in Elite Mountain Bikers: A Single-arm Pilot Study.

Mountain bike (MTB) racing is a highly intensive physical activity and requires a high degree of technical ability to perform at the elite athlete level, which might compromise mental well-being, increasing symptoms of anxiety and depression through overtraining, injury, and burnout. Yoga Pranayama is the key to bringing about psychosomatic integration and harmony. This study aimed to explore the effects of yoga pranayama practices (YPP) on elite mountain bikers' burnout. This is a single-arm pilot study. Twenty-seven subjects practiced 30 sessions of YPP seven times a week for 1 month. The outcomes measured were blood biochemical parameters accompanied by complete blood count and athlete burnout score. Cubital vein blood test and burnout questionnaire were conducted at baseline and after 1 month. Test results showed a significant decrease in cortisol (CO) (P = 0.001) and urea nitrogen (P < 0.001) and an increase in testosterone: CO ratio (P = 0.001). This study indicates that YPP might improve burnout in elite mountain bikers.

5-Azacytidine promotes HCC cell metastasis by up-regulating RDH16 expression.

The level of DNA methylation could affect the expression of tumor promoting and tumor suppressor genes. DNA methyltransferase inhibitors could reduce high methylation levels in cancer and inhibit the progression of a variety of cancers, including HCC. However, the pro-metastatic effect of DNA methyltransferase inhibitors in some cancers suggest the potential risk of their use. Whether DNA methyltransferase inhibitors also promote metastasis in HCC remains unclear. Our study will explore the effect of DNA methyltransferase inhibitor 5-Azacytidine on HCC metastasis. Our study found that 5-Azacytidine inhibited the proliferation of HCC cells while promoting in vitro and in vivo metastasis of HCC. Mechanistically, our study showed that 5-Azacytidine increased the expression of RDH16 by decreasing the methylation of RDH16 gene promoter. RDH16 is a highly methylated gene and its expression is very low in hepatocellular carcinoma. 5-Azacytidine promoted the migration of hepatocellular carcinoma cells by increasing the expression of RDH16. Our results suggest that 5-Azacytidine up-regulates the expression of RDH16 by decreasing the methylation level of RDH16, and then promoting HCC metastasis. These findings suggest that 5-Azacytidine and even other DNA methyltransferase inhibitors may have the risk of promoting metastasis in HCC treatment. RDH16 could be used as a pro-metastasis biomarker in the treatment of HCC with DNA methyltransferase inhibitors.

Trichinella spiralis cathepsin L induces macrophage M1 polarization via the NF-κB pathway and enhances the ADCC killing of newborn larvae.

BACKGROUND: During the early stages of Trichinella spiralis infection, macrophages predominantly undergo polarization to the M1-like phenotype, causing the host's inflammatory response and resistance against T. spiralis infection. As the disease progresses, the number of M2-type macrophages gradually increases, contributing to tissue repair processes within the host. While cysteine protease overexpression is typically associated with inflammation, the specific role of T. spiralis cathepsin L (TsCatL) in mediating macrophage polarization remains unknown. The aim of this study was to assess the killing effect of macrophage polarization mediated by recombinant T. spiralis cathepsin L domains (rTsCatL2) on newborn larvae (NBL). METHODS: rTsCatL2 was expressed in Escherichia coli strain BL21. Polarization of the rTsCatL2-induced RAW264.7 cells was analyzed by enzyme-linked immunosorbent assay (ELISA), quantitative PCR (qPCR), western blot, immunofluorescence and flow cytometry. The effect of JSH-23, an inhibitor of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), on rTsCatL2-induced M1 polarization investigated. Cytotoxic effects of polarized macrophages on NBL were observed using in vitro killing assays. RESULTS: Following the co-incubation of rTsCatL2 with RAW264.7 murine macrophage cells, qPCR and ELISA revealed increased transcription and secretion levels of inducible nitric oxide synthase (iNOS), interleukin (IL)-6, IL-1ß and tumor necrosis factor alpha (TNF-α) in macrophages. Western blot analysis showed a significant increase in iNOS protein expression, while the expression level of arginase-1 protein remained unchanged. Flow cytometry revealed a substantial increase in the number of CD86-labeled macrophages. The western blot results also indicated that rTsCatL2 increased the expression levels of phospho-NF-κB and phospho-nuclear factor-κB inhibitor alpha (IκB-α) proteins in a dose-dependent manner, while immunofluorescence revealed that rTsCatL2 induced nuclear translocation of the p65 subunit of NF-κB (NF-κB p65) protein in macrophages. The inhibitory effect of JSH-23 suppressed and abrogated the effect of rTsCatL2 in promoting M1 macrophage polarization. rTsCatL2 mediated polarization of macrophages to the M1-like phenotype and enhanced macrophage adhesion and antibody-dependent cell-mediated cytotoxicity (ADCC) killing of NBL. CONCLUSIONS: The results indicated that rTsCatL2 induces macrophage M1 polarization via the NF-κB pathway and enhances the ADCC killing of NBL. This study provides a further understanding of the interaction mechanism between T. spiralis and the host.

First ruthenium(II) polypyridyl complex as a true molecular "light switch" for triplex RNA structure: [Ru(phen)2(mdpz)]2+ enhances the stability of Poly(U)·Poly(A)*Poly(U).

Stabilization of triple helical structures is extremely important for carrying out their biological functions. Nucleic acid triple helices may be formed with DNA or RNA strands. In contrast to many studies in DNA, little has been reported concerning the recognition of the RNA triplex by transition-metal complexes. In this article, [Ru(phen)(2)(mdpz)](2+) (Ru1) is the first metal complex able to enhance the stability of the RNA triplex Poly(U)·Poly(A)*Poly(U) and serve as a prominent molecular "light switch" for the RNA triplex.

[Integration of soft and hard classifications using linear spectral mixture model and support vector machines].

This paper presents a new soft and hard classification. By analyzing the target objects in the image distribution, and calculating the adaptive threshold automatically, the image is divided into three regions: pure regions, non-target objects regions and mixed regions. For pure regions and non-target objects regions, hard classification method (support vector machine) is used to quickly extract classified results; For mixed regions, soft classification method (selective endmember for linear spectral mixture model) is used to extract the abundance of target objects. Finally, it generates an integrated soft and hard classification map. In order to evaluate the accuracy of this new method, it is compared with SVM and LSMM using ALOS image. The RMSE value of new method is 0.203, and total accuracy is 95.48%. Both overall accuracies and RMSE show that integration of hard and soft classification has a higher accuracy than single hard or soft classification. Experimental results prove that the new method can effectively solve the problem of mixed pixels, and can obviously improve image classification accuracy.

Knowledge about Human Papillomavirus and Cervical Cancer Prevention among Intern Nurses.

OBJECTIVE: Intern nurses will play an important role in the use of vaccination to prevent cervical cancer. This study assesses the knowledge about human papillomavirus (HPV) infection and cervical cancer prevention among intern nurses. METHODS: We developed a questionnaire to investigate intern nurses' knowledge about HPV infection and cervical cancer prevention. Participants included 323 intern nurses from eight schools. RESULTS: The effective response rate was 79.8%. Some (7.0%) knew that early-stage cervical cancer is commonly asymptomatic. Only 9.7% knew that infection is generally asymptomatic and 20.5% knew that vaccination has no major side effects. There were differences in gender, age, school type, and place of residence for several questions. CONCLUSIONS: This study indicates a low level of knowledge about HPV infection and cervical cancer prevention among intern nurses. Our findings highlight the need for more education in this topic to increase the knowledge of intern nurses.

Genome-wide RNAi Screening Identifies RFC4 as a Factor That Mediates Radioresistance in Colorectal Cancer by Facilitating Nonhomologous End Joining Repair.

PURPOSE: Neoadjuvant chemoradiotherapy (neoCRT) is a standard treatment for locally advanced rectal cancer (LARC); however, resistance to chemoradiotherapy is one of the main obstacles to improving treatment outcomes. The goal of this study was to identify factors involved in the radioresistance of colorectal cancer and to clarify the underlying mechanisms. EXPERIMENTAL DESIGN: A genome-wide RNAi screen was used to search for candidate radioresistance genes. After RFC4 knockdown or overexpression, colorectal cancer cells exposed to X-rays both in vitro and in a mouse model were assayed for DNA damage, cytotoxicity, and apoptosis. Moreover, the regulatory effects and mechanisms of RFC4 in DNA repair were investigated in vitro. Finally, the relationships between RFC4 expression and clinical parameters and outcomes were investigated in 145 patients with LARC receiving neoCRT. RESULTS: RFC4, NCAPH, SYNE3, LDLRAD2, NHP2, and FICD were identified as potential candidate radioresistance genes. RFC4 protected colorectal cancer cells from X-ray-induced DNA damage and apoptosis in vitro and in vivo. Mechanistically, RFC4 promoted nonhomologous end joining (NHEJ)-mediated DNA repair by interacting with Ku70/Ku80 but did not affect homologous recombination-mediated repair. Higher RFC4 expression in cancer tissue was associated with weaker tumor regression and poorer prognosis in patients with LARC treated with neoCRT, which likely resulted from the effect of RFC4 on radioresistance, not chemoresistance. CONCLUSIONS: RFC4 was identified as a radioresistance factor that promotes NHEJ-mediated DNA repair in colorectal cancer cells. In addition, the expression level of RFC4 predicted radiotherapy responsiveness and the outcome of neoadjuvant radiotherapy in patients with LARC.

[Effect of total flaveos of Gymostemma pentaphyllum on protein expression of Fas/FasL genes and TNF-alpha concentration in cultured neonatal rat cardiomyocytes with hypoxia-reoxygenation].

OBJECTIVE: To study the effect of total flaveos of Gymostemma pentaphyllum on the protein expression of apoptosis-associated Fas/FasL gene and tumor necrosis factor-alpha (TNF-alpha) concentration in cultured neonatal rat cardiomyocytes with hypoxia-reoxygenation (H/R). METHOD: A cultured primary neonatal rat cardiomyocytes model with H/R was erected, experiments were divided into six groups, (1)control group, (2)H/R group, (3)15 mg x L(-1) TFG plus H/R group, (4)45 mg x L(-1) TFG plus H/R group, (5) 105 mg x L(-1) TFG plus H/R group, (6)105 mg x L(-1) TFG group. TNF-aconcentration in cultured cardiomyocytes with H/R, was determined by ELISA method, the protein expression of Fas/FasL genes were estimated by immunohisto-chemistry. RESULT: After cardiomyocytes were made with H/R, Compared with control group, the positive expression index (PEI) of Fas/FasL proteins in cardiomyocytes increased significantly, Compared with H/R groups, the PEI of Fas/FasL proteins were lower significantly in groups with different dosages of TFG (P < 0.05). TFG inhibited the secretion of TNF-alpha from myocardial cells and increased the survival rate of myocardial cells. CONCLUSION: The protein expression of apoptosis-associated Fas/FasL genes increased during H/R. The TFG can protect myocardium against H/R injury by decreasing the production of TNF-alpha, downregulating the protein expression of Fas/FasL genes, and then inhibiting myocyte apoptosis.

[Therapeutic effect of BILT combined with praziquantel in treatment of chronic schistosomiasis].

OBJECTIVE: To investigate the clinical therapeutic effect of biological information infrared liver therapeutic apparatus (BILT) combined with praziquantel in the treatment of patients with chronic schistosomiasis. METHODS: A case-control study was conducted. A total of 142 chronic schistosomiasis patients were divided into an experimental group (BILT combined with praziquantel) with 64 cases and a control group (routine treatment with praziquantel alone) with 78 cases on the basis of the age, gender, disease duration and liver function as paired condition. Fatigue, diarrhea, abdominal distension, liver function, hyaluronic acid (HA) and laminin (LN) were as observation indexes and the observation results were compared between two groups. RESULTS: Before the treatment, there were no significant differences between the two groups in the indexes above-mentioned (P > 0.05). After the treatment, the incidence rates of fatigue, diarrhea, abdominal distension, abnormal liver function, and the levels of HA and LN in the experimental group were significantly lower than those in the control group (P < 0.01). CONCLUSIONS: BILT combined with praziquantel can significantly alleviate the short-term clinical symptoms, restore liver function and also alleviate hepatic fibrosis of the patients with chronic schistosomiasis.

Internal fixation vs conservative treatment for displaced distal radius fractures: a meta-analysis of randomized controlled trials.

BACKGROUND: The aim of the present study was to compare clinical outcomes of internal fixation and conservative approach in the treatment of displaced distal radius fractures. METHODS: Reports of studies were retrieved from the PubMed, Cochrane Library, EMBASE, BIOSIS, Ovid, CNKI, and Wanfang Data databases, as well as other sources. Methodological quality of the trials was critically assessed, and relevant data were extracted. Review Manager (RevMan) meta-analysis software (version 5.0; Cochrane Collaboration, London, UK) was used for data analysis. RESULTS: A total of 10 randomized controlled trials, which included 653 patients, were eligible for inclusion in the present meta-analysis, 7 of which were in English, and 3 of which were in Chinese. The trials had medium risk of bias. Results of meta-analysis showed that patients undergoing conservative treatment for distal radius fractures had better restoration of pronation (MD=1.80, 95% confidence interval [CI]=0.18-3.42, p=0.03; heterogeneity p=0.17, I2=43%), but shorter restoration of radial length (MD=2.62, 95% CI=1.47-3.76, p<0.00001; heterogeneity p=0.02, I2=73%). Wrist range of motion other than pronation, grip strength, radiographic parameters other than radial length, and rates of complications were not significantly different between the 2 treatments. CONCLUSION: Very few clinical differences were found between results of internal fixation and conservative treatment for displaced distal radius fractures. Best course of of treatment must be determined based on concrete conditions.

[Construction of recombinant human nerve growth factor (rh-ß-NGF) eukaryotic vector and its expression in HEK293 cells].

Human nerve growth factor (NGF) is a nerve cell growth regulation factor, which can provide nutrition for the neurons and promote the neurites outgrowth. In order to produce large-scale recombinant human nerve growth factor (rh-beta-NGF), we constructed a plasmid vector, which can stably express the rh-beta-NGF in the HEK293 cell lines. First, the plasmid of pCMV-beta-NGF-IRES-dhfr was constructed and transformed into HEK293 cells. Then MTX pressurized filter and limiting dilution methods were used to obtain monoclonal HEK293 cell lines. After stepwise reducing serum in culture media, the cells eventually adapted to serum-free medium and secreted rh-beta-NGF. SDS-PAGE analysis revealed that the expression product owned a molecular weight of about 13 kDa and a purity of more than 50%. The peptide mapping sequencing analysis demonstrated the sequences of rh-beta-NGF matched with the theoretical ones. Later we purified this protein by ion exchange and molecular sieve chromatograph. Finally, our experimental results exhibited that the recombinant cell lines can stably express rh-beta-NGF with a high efficiency of more than 20 pg/cell x day. In addition, this protein could successfully induce differentiation of PC12 cells. In summary, our recombinant HEK293 cells can express bio-active rh-beta-NGF with great efficiency and stability, which supply a valid basis to large-scale production of rh-beta-NGF.

Heme oxygenase-1 mediated memorial and revivable protective effect of ischemic preconditioning on brain injury.

AIMS: Ischemic preconditioning (IPC) has short-term benefits for stroke patients. However, if IPC protective effect is memorial and the role of the intracellular protective protein heme oxygenase-1 (HO-1) is not known. METHODS: Ischemic preconditioning and the corresponding sham control were achieved by blocking the blood flow of the left internal carotid artery for 20 min and 2 second, respectively, in rats. Both IPC and sham-operated animals were divided into three groups and treated with PBS, the HO-1 inducer hemin, and the HO-1 inhibitor Znpp. Three weeks after IPC, brain ischemia-reperfusion injury was achieved by left middle cerebral artery obstruction for 45 min followed by 24-h reperfusion. RESULTS: 2,3,5-triphenyltetrazolium chloride staining and neurological dysfunction scoring showed IPC significantly reduced brain infarct area and improved neurological function occurred 3 weeks after IPC. Hemin treatment promoted whereas ZnPP blocked the benefits of IPC. Immunohistochemical analysis and Western blotting showed that the expression of HO-1 was higher in the border zone than in the necrotic core zone. The memorial IPC protection is independent of adenosine receptor A1R and A2aR expressions. CONCLUSIONS: We found for the first time that the protective effect of IPC can be remembered to protect brain injury occurred after acute response disappear. The results indicate that interventional treatment can achieve protective effect for future cerebral injury not only through interventional treatment itself but also through the memorial and revivable IPC, eliminating the concern that temporary ischemia caused by interventional treatment may leave harmful effect in the brain.

Autophagy is induced through the ROS-TP53-DRAM1 pathway in response to mitochondrial protein synthesis inhibition.

Mitoribosome in mammalian cells is responsible for synthesis of 13 mtDNA-encoded proteins, which are integral parts of four mitochondrial respiratory chain complexes (I, III, IV and V). ERAL1 is a nuclear-encoded GTPase important for the formation of the 28S small mitoribosomal subunit. Here, we demonstrate that knockdown of ERAL1 by RNA interference inhibits mitochondrial protein synthesis and promotes reactive oxygen species (ROS) generation, leading to autophagic vacuolization in HeLa cells. Cells that lack ERAL1 expression showed a significant conversion of LC3-I to LC3-II and an enhanced accumulation of autophagic vacuoles carrying the LC3 marker, all of which were blocked by the autophagy inhibitor 3-MA as well as by the ROS scavenger NAC. Inhibition of mitochondrial protein synthesis either by ERAL1 siRNA or chloramphenicol (CAP), a specific inhibitor of mitoribosomes, induced autophagy in HTC-116 TP53 (+/+) cells, but not in HTC-116 TP53 (-/-) cells, indicating that tumor protein 53 (TP53) is essential for the autophagy induction. The ROS elevation resulting from mitochondrial protein synthesis inhibition induced TP53 expression at transcriptional levels by enhancing TP53 promoter activity, and increased TP53 protein stability by suppressing TP53 ubiquitination through MAPK14/p38 MAPK-mediated TP53 phosphorylation. Upregulation of TP53 and its downstream target gene DRAM1, but not CDKN1A/p21, was required for the autophagy induction in ERAL1 siRNA or CAP-treated cells. Altogether, these data indicate that autophagy is induced through the ROS-TP53-DRAM1 pathway in response to mitochondrial protein synthesis inhibition.

SC(3): Triple spectral clustering-based consensus clustering framework for class discovery from cancer gene expression profiles.

In order to perform successful diagnosis and treatment of cancer, discovering, and classifying cancer types correctly is essential. One of the challenging properties of class discovery from cancer data sets is that cancer gene expression profiles not only include a large number of genes, but also contains a lot of noisy genes. In order to reduce the effect of noisy genes in cancer gene expression profiles, we propose two new consensus clustering frameworks, named as triple spectral clustering-based consensus clustering (SC3) and double spectral clustering-based consensus clustering (SC2Ncut) in this paper, for cancer discovery from gene expression profiles. SC3 integrates the spectral clustering (SC) algorithm multiple times into the ensemble framework to process gene expression profiles. Specifically, spectral clustering is applied to perform clustering on the gene dimension and the cancer sample dimension, and also used as the consensus function to partition the consensus matrix constructed from multiple clustering solutions.Compared with SC3, SC2Ncut adopts the normalized cut algorithm, instead of spectral clustering, as the consensus function.Experiments on both synthetic data sets and real cancer gene expression profiles illustrate that the proposed approaches not only achieve good performance on gene expression profiles, but also outperforms most of the existing approaches in the process of class discovery from these profiles.

[Spatial relevance between natural regeneration of Picea and heterogeneity of soil available nitrogen in Guandi Mountain].

By using geostatistic and pattern analysis methods, this paper studied the spatial pattern of Picea seedlings in naturally regenerated conifer (Picea) and mixed (Picea-Populus-Betula) forests in Guandi Mountain of Shanxi Province, China. The spatial distribution of soil nitrogen was also quantified by semivariogram analysis. To understand the effects of spatial heterogeneity of soil nitrogen on the regeneration of Picea seedlings, the relationships between the regeneration pattern of the seedlings and the spatial distribution of soil nitrogen were investigated by using GIS superposition and statistical analysis. In conifer stands, the distribution of Picea seedlings appeared as a patch pattern and was auto-correlated; while in mixed stands, the distribution was of gathering distribution pattern controlled by random factors. In the Picea stands with relatively low soil nitrogen content, the spatial distribution of soil available nitrogen was significantly heterogeneous and auto-correlated; whereas in the mixed stands with high nitrogen content, the distribution of soil available nitrogen showed random heterogeneity. In the conifer stands, the spatial correlation between Picea seedlings regeneration pattern and soil available nitrogen distribution was significant, regenerating more seedlings in the patches with higher NH4(+) -N concentration; while in the mixed stands, the correlation was not significant.

[Neighborhood relationships of land use spatial pattern in Qixia City].

By using the modules of neighborhood statistics and spatial analysis in ArcGIS software, an analysis was conducted with neighborhood factors on the spatiotemporal variation trend of the neighborhood relationships among main land use types in Qixia City of Shandong Province from 1987 to 2003. The results indicated that the neighborhood relationships between different land use types increased with increasing neighborhood distance, while those among the same land use types manifested congregation, which decreased with increasing neighborhood distance. From 1987 to 2003, the neighborhood relationships between construction land and orchard land, and between woodland and orchard land presented decreasing trend, while those of construction land with woodland and grassland were in adverse. Some measures such as controlling construction land area, protecting cultivated land, and decreasing the disturbances of human activities on woodland and grassland should be carried out to realize the harmonious development of economic and ecological benefits in Qixia City.

[Effects of sinomenine on intracellular free calcium concentration and the activity of protein kinase in cultured rabbit aortic smooth muscle cells].

AIM: To explore the effects of sinomenine(Sin) on intracellular free calcium ([Ca2+]i) and the activity of PKC (protein kinase C) of the cultured aortic vascular smooth muscle cells (VSMC) during ischemia and hypoxia. METHODS: The effect of Sin on changes in [Ca2+]i were determined in cultured rabbit VSMC after exposure to high K+, norepinephrine (NE) and caffeine (Caf). Fluorescent Ca2+ -indicater fura-2/AM was used. The effects of Sin were compared with that of verapamil (Ver). The hypoxia model was made, then the activity of PKC was measured by y scintillation counting instrument. RESULTS: Sin (10 x 10(-6) mol x L(-1), 3 x 10(-5) mol x L(-1) 10(-4) mol x L(-1)) inhibited the elevation of [Ca2+]i induced by high K+ -depolarization in a concentration dependent manner. In addition, Sin inhibited the elevation of [Ca2+]i induced by NE in the presence of extracellular Ca2+. In the absence of extracellular Ca2+, Sin (3 x 10(-5) mol.L(-1)) also had no blocking effect on the NE-induced [Ca2+]i increase. It was found that the activity of PKC treated with Sin in VSMC cytoplasm and cell membrane in normal condition increased, the activity of PKC in cytoplasm in ischemia and hypoxia situation increased, but the activity of PKC in cell membrane decreased. When VSMC was treated with Sin, the activity of PKC in cytoplasm decreased and that of cell membrane increased. CONCLUSION: The results suggest that Sin might decrease the[Ca2+] i of VSMC by blocking both VDC and ROC, could regulate the PKC activities induced by ischemia and hypoxia.