Poly(U)-binding-splicing factor 60 KDa (PUF60), a U2-related splicing factor that facilitates 3′splice-site recognition at the early stage of spliceosome assembly.High expression of PUF60 is related to the occurrence of multiple tumors, such as colorectal cancer, ovarian cancer, gastric cancer, liver cancer, et al.PUF60 can regulate the expression of c-myc through the core-human transcription factor Ⅱ basal transcription factor.Anti-PUF60 antibodies are detected in the sera of patients with early-stage and recurrent tumor, and the levels are significantly decreased after the operation.PUF60 can be used as a combined or independent test index for the diagnosis of cancer, which can be a potential new target for gene therapy.

Objective To discuss the clinical application of medical coordination and hierarchical responsibility system in interventional nursing care which is carried out based on the “Henan Province Model”. Methods From November 2003 to April 2014 at authors’ hospital, medical coordination and hierarchical responsibility system was executed through setting up a ranking system of nursing position , optimizing shift process , dividing physician-nurse medical liability groups , strengthening medical training , implementing medical coordination service mode, etc. The clinical results were compared with those of conventional nursing care that were recorded during the period from May to Oct. of 2013. Results After implementation of medical coordination and hierarchical responsibility system, the quality of medical care, the patent’s satisfaction, the cooperation satisfaction of physicians and nurses were significantly improved. Compared with those before implementation of medical coordination and hierarchical responsibility system , the differences in the above indexes were statistically significant(P<0.01 or P<0.05). Conclusion Based on multidisciplinary joint diagnosis and treatment, and combined with coordination physician-nurse service mode, the enthusiasm of nursing staff can be motivated and the tacit understanding between physicians and nurses as well as the quality of nursing service can be improved. All of the above will promote the development of nursing discipline.

AIM:To investigate the expression of poly (ADP-ribose) polymerase-1 (PARP-1) in the epithelial ovarian cancer ( EOC) and its relationship with epithelial-mesenchymal transition ( EMT) .METHODS:The expression of PARP-1, E-cadherin, vimentin and Snail was detected in the EOC and benign ovarian tumor tissues by immunohistochemi -cal method and real-time PCR.The expression of PARP-1, E-cadherin, vimentin and Snail proteins in the SKOV3 cells treated with efficient PARP-1 inhibitor PJ34 was determined by Western blotting .RESULTS:The positive expression rates of PARP-1, vimentin and Snail were significantly higher in the EOC than that in the benign ovarian tumor tissues , whereas the positive expression rate of E-cadherin was the opposite (P<0.05).The expression of PARP-1, E-cadherin, vimentin and Snail in the EOC was associated with the histological grade , clinical stage and lymphatic metastasis (P<0.05), but no relationship with age and pathological types was observed .The expression of E-cadherin in the EOC was negatively co-related to that of PARP-1.In contrast, the expression of vimentin and Snail in the EOC was positively co-related to that of PARP-1.The relative mRNA expression of PARP-1, vimentin and Snail in the EOC was significantly higher than that in the benign ovarian tumor tissues ( P<0.05) , while the mRNA expression of E-cadherin in the EOC was remarkably lower than that in the benign ovarian tumor tissues (P<0.05).The protein expression of PARP-1, vimentin and Snail in the SKOV3 cells was significantly decreased (P<0.05), while E-cadherin protein was increased after treated with PJ 34(P<0.05). CONCLUSION:PARP-1 may contribute to the onset of EMT in the EOC by regulating the expression of E -cadherin, vim-entin and Snail.The role of PARP-1, which is relevant to EMT, might be important in the development of ovarian cancer .

Objective To construct HaCaT cell lines stably expressing the wild type human GJB6 gene or its mutant by using a Tet-On lentiviral vector, and to lay an experimental foundation for studies on pathogenesis of hidrotic ectodermal dysplasia. Methods The wild-type human GJB6 gene and its mutant (A88V)were amplified by PCR, and then inserted into the Tet-on lentivirus plasmid to construct recombinant lentivirus vectors. The recombinants were identified by gene sequencing and enzymatic digestion. Cultured HaCaT cells were classified into three groups to be transfected with a negative control lentiviral vector (NC group), the lentivirus vector expressing the wild-type human GJB6 gene (WT group), or the lentivirus vector expressing the mutant human GJB6 gene (MU group). Puromycin was used to select HaCaT cell clones stably expressing the GJB6 gene which encodes the connexin 30 (Cx30)protein. The selected HaCaT cell clones were cultured with or without tetracycline for 48 hours, thereafter, real-time PCR(RT-PCR) was performed to detect GJB6 gene mRNA expression, Western-blot analysis to measure expressions of Cx30 and FLAG-tag proteins, and cell counting kit 8 (CCK8)assay to evaluate cellular proliferative activity. Results Enzymatic digestion and gene sequencing showed that recombinant lentivirus plasmids were successfully constructed. RT-PCR showed evidently increased mRNA expression of the GJB6 gene in stably transfected HaCaT cells. Moreover, the expression abundance of the GJB6 gene was 112.369 times higher in the WT group induced by tetracycline than in that without tetracycline treatment (P < 0.05), and 2.249 times higher in the MU group induced by tetracycline than in that without tetracycline treatment (P < 0.05). Western-blot analysis showed that Cx30 and FLAG-tag proteins were stably expressed in the WT group and MU group after induction with tetracycline, while neither of them was observed in the WT group or MU group without tetracycline treatment, or in the NC group. Significant differences were noted in cellular proliferative activity (expressed as the absorbance value at 450 nm)between the MU group with and without tetracycline treatment and between the WT group with and without tetracycline treatment at 4, 8, 12, 24, 36 and 48 hours (all P <0.05), but not between the NC group with and without tetracycline treatment at any of the above time points (all P >0.05). Conclusion HaCaT cell lines which stably express the wild-type GJB6 gene or its mutant(A88V)are successfully constructed.

An electrochemical sensor incorporating a signal enhancement for the determination of lead (Ⅱ) ions (Pb2+) was designed on the basis of the thrombin-binding aptamer (TBA) as a molecular recognition element and ionic liquid supported cerium oxide (CeO2) nanoparticles-carbon nanotubes composite modification. The composite comprises nanoparticles CeO2, multi-wall carbon nanotubes (MWNTs)and hydrophobic room temperature ionic liquid (RTIL) l-ethyl-3-methylimidazolium tetrafluoroborate (EM1MBF4). The electrochemical sensors were fabricated by immersing the CeO2-MWNTs-EMIMBF4 modified glassy carbon electrode (GCE) into the solution of TBA probe. In the presence of Pb2+, the TBA probe could form stable G-quartet structure by the specific binding interactions between Pb2+ and TBA. The TBA-bound Pb2+ can be electrochemically reduced, which provides a readout signal for quantitative detection of Pb2+. The reduction peak current is linearly related to the concentration of Pb2+ from 1.0 × 10 8 M to 1.0 × 10-5 M with a detection limit of 5 × 109 M. This work demonstrates that the CeO2-MWNTs-EMIMBF4 nanocomposite modified GCE provides a promising platform for immobilizing the TBA probe and enhancing the sensitivity of the DNA-based sensors.

The expensive production of bioethanol is because it has not yet reached the 'THREE-HIGH' (High-titer, high-conversion and high-productivity) technical levels of starchy ethanol production. To cope with it, it is necessary to implement a high-gravity mash bioethanol production (HMBP), in which sugar hydrolysates are thick and fermentation-inhibitive compounds are negligible. In this work, HMBP from an atmospheric glycerol autocatalytic organosolv pretreated wheat straw was carried out with different fermentation strategies. Under an optimized condition (15% substrate concentration, 10 g/L (NH4)2SO4, 30 FPU/g dry matter, 10% (V/V) inoculum ratio), HMBP was at 31.2 g/L with a shaking simultaneous saccharification and fermentation (SSF) at 37 degrees C for 72 h, and achieved with a conversion of 73% and a productivity of 0.43 g/(L x h). Further by a semi-SFF with pre-hydrolysis time of 24 h, HMBP reached 33.7 g/L, the conversion and productivity of which was 79% and 0.47 g/(L x h), respectively. During the SSF and semi-SSF, more than 90% of the cellulose in both substrates were hydrolyzed into fermentable sugars. Finally, a fed-batch semi-SFF was developed with an initial substrate concentration of 15%, in which dried substrate (= the weight of the initial substrate) was divided into three portions and added into the conical flask once each 8 h during the first 24 h. HMBP achieved at 51.2 g/L for 72 h with a high productivity of 0.71 g/(L x h) while a low cellulose conversion of 62%. Interestingly, the fermentation inhibitive compound was mainly acetic acid, less than 3.0 g/L, and there were no other inhibitors detected, commonly furfural and hydroxymethyl furfural existing in the slurry. The data indicate that the lignocellulosic substrate subjected to the atmospheric glycerol autocatalytic organosolv pretreatment is very applicable for HMBP. The fed-batch semi-SFF is effective and desirable to realize an HMBP.
Biofuels ; Carbohydrates ; chemistry ; Cellulose ; chemistry ; Ethanol ; metabolism ; Fermentation ; Furaldehyde ; chemistry ; Glycerol ; chemistry ; Hydrolysis ; Triticum

A real-time polymerase chain reaction ( PCR ) micro total analysis system (μ-TAS ) integrated nucleic acid extraction, PCR amplification and real-time-fluorescent PCR detection on a same microfluidic chip was prepared for the fully automated and on-chip analysis of nucleic acid. The proposed method had the advantage such as low sample consumption, fast analysis and simple operation and so on. Micromachining technology was used to fabricate the anodic molds of integrated nucleic acid extraction microfluidic chip. A polydimethylsiloxane (PDMS) substrate with 3D channels was manufactured by a combination of molds and an injection molding method. The glass substrate and the chip were bonded together using a plasma treatment. The μ-TAS included a microfluidic control device whose micro fluidic velocity ( 0-10 mL/min ) could be adjusted, a TEC platform which the precision of temperature control was 0. 1℃ and a CCD detection module. The DNA of human blood was extracted by using a silica gel membrane method on the microfluidic chip. The processes of DNA extraction and detection were preset in the μ-TAS. Human blood lysate ( 20 μL ) were driven to the extraction chamber and was then washed. The fluidic drive speed was 2 mL/min. DNA and PCR reagents were mixed and then were driven into the PCR chamber. The fluidic drive speed was 1 mL/min. The GAPDH gene in extracted genome DNA was amplified by PCR and detected. The amplified product was verified by melting analysis. The results of nucleic acid extraction method on the chip were compared to those obtained using a standard manual centrifuge extraction method. The amplification curves were obvious. Ct values of the chip method were 25 . 3 and 26 . 9 . The denaturation temperature of all the melting was 89 . 9 ℃. The results validated that the chip-based method and device realized the extraction of nucleic acid, amplification and detection automatically.

Objective:To explore the impact of celastrol(CEL)on angiogenesis in rats with endometriosis(EM)and its regu-latory mechanism on Hippo-YAP signal pathway.Methods:SD female rats were grouped into six groups:sham operation group,model group,positive drug group(gestrinone),Verteporfin group(YAP inhibitor),low and high doses CEL groups,with 12 rats in each group.The weight and volume of rat endometrium were measured;HE staining was applied to observe the histological changes of rat endometrium;the microvessel density(MVD)in rat endometrium was detected by immunohistochemistry;the expressions of vascular endothelial growth factor(VEGF),TNF-α,IL-1β and IL-6 in rat serum were detected by ELISA;Western blot was applied to detect the expressions of VEGF and YAP proteins in rat endometrium.Results:Compared with sham operation group,the EM rats in model group showed obvious ectopic endometrial tissue lesions,increased number of interstitial cells,and obvious vascular proliferation,the weight,volume,MVD,the contents of VEGF,TNF-α,IL-1β,IL-6,and the expression levels of YAP,TAZ,and VEGF receptor 2(VEGFR2)proteins in endometrial tissue increased(P<0.05);compared with model group,the ectopic endometrium tissue of EM rats in positive drug group,Verteporfin group and low and high dose CEL groups gradually fell off,the number of interstitial cells were decreased,the structure was loose,and the blood vessels were significantly reduced,the weight,volume,MVD,the contents of VEGF,TNF-α,IL-1β,IL-6,and the expression levels of YAP,TAZ,and VEGFR2 proteins in endometrial tissue were decreased(P<0.05).Conclusion:CEL can inhibit angiogenesis in EM rats by inhibiting Hippo-YAP signal pathway.

Objective:To explore the effect of the etoposide (VP-16) on the reactivation of human immunodeficiency virus (HIV-1) from latent infection and study its potential molecular mechanism.Methods:The HIV-1 latently infected cell line J-Lat was treated with DMSO, VP-16 and vorinostat (SAHA), flow cytometry was used to detect the positive ratio of green fluorescent protein (GFP) in J-Lat, which can reflect the efficacy of high concentration VP-16 on reactivation. Then the reactivation efficiency of VP-16 on HIV-1 latently infected cells at different concentration and treatment time was measured by flow cytometry. The expression of CD25, CD69 and interleukin-6 (IL-6) of the VP-16-treated J-Lat cells were also detected in the same way. The effect of VP-16 on the transcription of long terminal repeat (LTR) was tested using a dual-luciferase reporter assay. The protein expression of the silent information regulator 1 (SIRT1) and the acetylated nuclear factor κB (NF-κB) p65 under the effect of VP-16 was checked by western blot (WB). Lentivirus-mediated SIRT1 short-hairpin RNA (SIRT1-shRNA) was employed to knock down SIRT1, and then tested for efficiency of reactivation.Results:The results of flow cytometry showed that VP-16 specifically reactivated HIV-1 latently infected J-Lat cells in a concentration and time-dependent manner. Meanwhile, the expression of CD25 and CD69 was upregulated to a certain extent, but the expression of IL-6 was not affected. Dual-luciferase reporter assay suggested that VP-16 positively regulated the transcription of HIV-1 LTR through NF-κB signaling pathway. WB results indicated that VP-16 can inhibit the protein expression of SIRT1 and promote the acetylation of NF-κB p65. Lentivirus-mediated SIRT1-shRNA successfully knocked down the mRNA and protein expression of SIRT1, and reactivated the HIV-1 latency effectively.Conclusions:VP-16 upregulates the acetylation of NF-κB p65 by inhibiting the expression of SIRT1, which subsequently promotes the transcription of HIV-1 LTR and reactivates the latent HIV-1 infection.

Objective This article reviews the curriculum framework, teaching content, teaching method and application effects of stimulation teaching of hospice care for nursing undergraduates and reveals its inspirations to China, in order to provide a reference for the further development of China's hospice education.