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.

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.

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 .

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 observe the effects of fecal microbiota transplantation on intestinal microbiota and brain function in sepsis rats. Methods Sixty male Sprague Dawley (SD) rats were divided into sham operation group, model group and fecal microbiota transplantation (FMT) group by random number table, each group 20 rats. The rat model of sepsis was established by injection of lipopolysaccharide (LPS) 10 mg/kg in tail vein. FMT group received nasogastric infusion of feces from healthy donor. Fecal samples were collected on the 6th day after the modeling to detect the levels of intestinal microbiota composition; the brain function was also evaluated by electroencephalogram (EEG), and the proportion of each waveform in EEG was calculated. After sacrifice of rats in different groups, the brain tissues were taken, the levels of protein expression and positive cells of Iba-1 in brain tissue were detected by Western Blot and immunohistochemistry method. Results ① Intestinal flora analysis showed that: the diversity index and Chaol index of the intestinal microbiota in model group were significantly lower than that in sham operation group (observed species:282±40 vs. 473±37, Chao1 index: 730±21 vs. 837±27, both P < 0.05); compared with the model group, the diversity index and Chaol index in FMT group were obviously higher (observed species: 461±20 vs. 282±40, Chao1 index:840±16 vs. 730±21, both P < 0.05). At phylum, family, genus level analysis showed that the proportion of Firmicutes phylum and Fusobacterium were obviously lower than those of sham operation group [Firmicutes phylum (22.12±1.34)% vs. (78.01±1.23)%, Fusobacterium: (2.03±0.17)% vs. (5.03±0.19)%, both P < 0.05], and the proportions of Proteobacteria, Bacteroidetes phyla and Acidaminococcaceae, Fusobacteriaceae, Enterbacteriacecae, Alistipes were markedly higher in model group [Proteobacteria: (70.21±2.35)% vs. (19.45±2.17)%, Bacteroidetes phyla: (4.12±0.19)% vs. (2.50±0.64)%, Acidaminococcaceae: (12.51±0.87)% vs. (1.01±0.12)%, Fusobacteriaceae: (13.62±1.27)% vs. (2.31±0.19)%, Enterbacteriacecae: (18.24±2.13)% vs. (4.15±1.51)%, Alistipes: (4.53±0.27)% vs. (1.47±0.33)%, all P < 0.05]; compared with the model group, the proportion of Firmicutes phylum and Faecalibacterium in FMT group were significantly higher [Firmicutes phylum: (72.14±2.31)% vs. (22.12±1.34)%, Faecalibacterium: (5.01±0.27)% vs. (2.03±0.17)%, both P < 0.05], and Proteobacteria, Bacteroidetes phyla and Acidaminococcaceae, Fusobacteriaceae, Enterbacteriacecae in FMT group were obviously lower [Proteobacteria: (14.23±1.98)% vs. (70.21±2.35)%, Bacteroidetes phyla: (3.15±0.18)% vs. (4.12±0.19)%, Acidaminococcaceae: (0.91±0.11)% vs. (12.51±0.87)%, Fusobacteriaceae: (1.25±0.15)% vs. (13.62±1.27)%, Enterbacteriacecae: (3.50±0.21)% vs. (18.24±2.13)%, all P < 0.05]. ② EEG analysis showed that the percentages of δ wave in EEG in model group was significantly higher after modeling than that in sham operation group [(16.86±0.50)% vs. (10.67±0.65)%, P < 0.05]; the ratios of δ wave in EEG was significantly lower in FMT group than that in the model group [(12.87±0.60)% vs. (17.35±0.41)%, P <0.05]. The incidence of abnormal EEG in sham operation group was 0, the incidence of abnormal EEG in model group was significantly increased [the ratios of δpredominant wave, θpredominant wave, low-voltage were 66.7% (6/9), 66.7% (6/9), 77.8% (7/9) respectively], the ratios of above abnormal waves in EEG in FMT group were obviously lower than those in model group [the ratios of above abnormal waves in FMT group were respectively 9.1% (1/11), 9.1% (1/11), 18.2%(2/11)]. ③ Western Blot analysis showed that the protein expression of Iba-1 in cortex in model group obviously was higher than that in sham operation group (Iba-1/β-actin: 1.39±0.16 vs. 0.67±0.18, P < 0.05); the expression of Iba-1 in cortex tissue of FMT group was markedly lower than that in model group (Iba-1/β-actin: 0.51±0.14 vs. 1.39±0.16, P < 0.05). ④ Immunohistochemistry of Iba-1 in cortex analysis showed that there were no Iba-1 positive cells in the cortex in sham operation group; Iba-1 positive cells were found in the cortex in model group; the number of Iba-1 positive cells in FMT group was less than that in model group. Conclusion FMT can improve the construction of intestinal microbiota, and ameliorate the brain dysfunction in SAE.

Objective:To analyze the clinical characteristics of bone tumors in children, so as to improve the understanding and diagnosis of bone tumors in children.Methods:The clinical data of bone tumors in children hospitalized in the Affiliated Hospital of Zunyi Medical University from January 2009 to December 2018 were collected, with the age ≤ 14 years old.All children′s gender, age, tumor type, location, number of lesions and first symptoms were counted, and the clinical characteristics of bone tumors in children in this area were analyzed.Results:Totally 548 children with bone tumor were collected, with 344 males and 204 females, and the ratio of males to females was 1.69∶1.00, with 462 single cases and 86 multiple cases.Among the multiple cases (86 cases), 82 cases (95.35%) were benign bone tumors.The total number of cases increased with age.The age of different tumors has its own characteristics.Tibia was the most common tumor location (223 in total), followed by femur (177 in total) and humerus (82 in total). According to the classification of tumor nature, there were 478 cases (87.22%) of benign bone tumors, 43 cases (7.85%) of intermediate bone tumors and 27 cases (4.93%) of malignant bone tumors.The incidence of benign bone tumors was significantly higher than that of intermediate and malignant bone tumors, with statistically significant differences ( χ2=72.604, P<0.05). Among the benign bone tumors (478 cases), osteochondroma was the most common (265 cases), accounting for 55.44%; among the intermediate bone tumors (43 cases), aneurysmal bone cyst was the most common (20 cases), occupying 46.51%; among the malignant bone tumors(27 cases), osteosarcoma was the most common (16 cases), accounting for 59.26%.Among the initial symptoms, 268 cases were local masses, and 166 cases were pain, which were significantly higher than that of 79 without symptoms, with 28 lameness, 4 deformity and 3 dysfunction, and the differences were statistically significant( χ2=71.422, P<0.05). Conclusions:The majority of bone tumors patients in children were male, and the incidence increased with age.Benign and solitary are common, and the multiple cases were mainly benign.The most common locations are tibia and femur.The initial symptoms were mainly local mass and pain.Most importantly, the clinical characteristics of different tumors are slightly different.

Objective To analyze the clinical characteristics,imaging manifestations and prognoses of anti-GABAB receptor antibodies encephalitis.Methods The clinical manifestations,laboratory findings and radiological data of 13 patients with anti-GABAB receptor encephalitis,admitted to our hospital from September 2015 to March 2017,were retrospectively analyzed.Modified Rankin scale (mRs) was performed to evaluate the prognoses (good prognosis:mRs scores ＜ 2;poor prognosis mRs scores≥3).Results These 13 patients had an average age of 58 years (ranged 49-76 years) with a male to female ratio of 12:1.The major clinical features,including epileptic seizure,were found in 12 patients,psychiatric symptoms in 11 patients,cognitive disorder in 7 patients,and disturbance of consciousness in 4 patients.Brain MR imaging showed abnormal signal in 5 patients:4 were located in the hippocampus and amygdaloid,and one in the pons and left temporal lobe.Five patients showed abnormalities in PET-CT,including 4 with temporal hypermetabolism and 1 with cortical hypometabolism.Chest CT showed lung occupying lesions in 4 patients,of which 2 patients were diagnosed as having small cell lung cancer (SCLC) by pathological examination.Ten patients received immunomodulatory therapy,and three were with supportive care.After the average of 8 months of follow-up,7 patients had good prognosis,5 patients had poor prognosis and one patient lost of follow up.Conclusions Anti-GABAB receptor encephalitis frequently occurs in elderly male subjects and the main characteristic includes prominent refractory epilepsy and shows neurological improvement on immunotherapy.It can accompany by SCLC and have a relatively poor prognosis.