Objective:To investigate the expression of microRNA-324-5p (miR-324-5p) and transcription factor forkhead box C1 (FOXC1) in glioma and their relationship with the prognosis of patients.Methods:From March 2012 to March 2015, a total of 72 cases of glioma tissues were collected from glioma patients who were admitted to Chongqing Hygeia Tumor Hospital and the People's Hospital of Nanchuan in Chongqing, and 28 cases of normal human brain tissues resected in craniocerebral surgery were also collected. The expressions of miR-324-5p and FOXC1 mRNA were detected by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR), and the expression of FOXC1 protein was detected by immunohistochemistry. Pearson method was used to analyze the correlation between the expressions of miR-324-5p and FOXC1 in glioma tissues; Kaplan-Meier method was used to analyze the survival of patients with glioma; Cox regression analysis was used to analyze the risk factors affecting the prognosis of patients with glioma.Results:FOXC1 protein was mainly located in the cytoplasm of glioma, and its positive expression rate in glioma tissues was 81.94% (59/72), which was significantly higher than that in normal brain tissues [17.86% (5/28)], and the difference was statistically significant ( χ2 = 35.938, P<0.01). Compared with normal brain tissues, the expression of miR-324-5p was down-regulated in glioma tissues (0.62±0.19 vs. 0.98±0.02, t = 9.974, P < 0.05), and the expression of FOXC1 mRNA was up-regulated (1.41±0.29 vs. 0.99±0.02, t = 7.633, P < 0.05). The expressions of miR-324-5p and FOXC1 protein were correlated with the number of primary lesions, differentiation degree, TNM stage and lymph node metastasis of glioma (all P<0.05). Pearson analysis showed that the expressions of miR-324-5p and FOXC1 mRNA were negatively correlated ( r = -0.550, P<0.01). The 5-year overall survival rate of patients in miR-324-5p high-expression group was significantly higher than that of patients in miR-324-5p low-expression group (45.71% vs. 24.33%, χ2 = 6.531, P = 0.011), and the 5-year overall survival rate of patients in FOXC1 protein high-expression group was significantly lower than that of patients in FOXC1 protein low-expression group (30.41% vs. 42.34%, χ2 = 3.631, P = 0.047). Multivariate Cox regression analysis showed that low differentiation, TNM stage Ⅲ-Ⅳ, lymph node metastasis, low expression of miR-324-5p and high expression of FOXC1 protein were independent risk factors for prognosis of glioma patients (all P < 0.05). Conclusions:The expression of miR-324-5p is low and the expression of FOXC1 is high in glioma. They may be involved in the regulation of tumor differentiation and metastasis, and related to the poor prognosis of patients. They may be potential therapeutic targets for glioma.

Objective To discuss the technique and value of preoperative reformatting with three-dimensional computed tomography technique for C_2 pedicle screw track.Methods GE Light Speed 16 Pro spinal CT scans of 15 adult dry vertebrae were loaded into an imaging station (software ADW4.2).Two methods of C_2 pedicle screw techniques were analyzed through virtual screw trajectory by VR (volume rendering) and MPR (multiple planar reformatting) techniques,in method A,screw entry point was the intersection between the media-vertical and the cranial line of C_2 inferior facet joint,in method B,the screw track was from the cranial and medial quadrant of the dorsal part of C_2 inferior facet joint.Results The screw track could be observed dynamically from any plane.Two vertebrae were ob- served with smaller height in isthmus and the medial edge of the transverse foramen since no space was a- vailable for the screw.The screw trajectories data were compared between method A and method B,which showed that the angles towards the cephalad (in sagittal plane) and midline (in transverse plane) were bigger in method A than in method B (P＜0.05,0.01),but the safe screw diameter was smaller in method A than in method B (P＜0.05),and there was no difference of the screw length between the two methods(P＜0.05 ).Conclusion In this research,the individual C_2 pedicle screw entry points, screw diameter and security screw angle can be simulated,and the screw track can be observed dynami- cally to make sure if it transits the bone structure completely.Preoperative three-dimensional computed tomography reformatting for pedicle screw track is of great value in clinical and basic researches.

Propolis is an adhesive substance mixed with plant resins collected by honeybees (Apis mellifera) and secretions from their mandibular gland and wax gland, with wide pharmacological activity and healthcare functions. Its antioxidant activity has long been regarded as one of the most important biological activities of propolis. This article summarizes studies on the antioxidant activity of propolis extracts from different geographic origins and with different extraction methods, as well as several important monomer active ingredients in propolis, and concludes the potential molecular mechanism of antioxidant activity of propolis and its monomer ingredients, with the aim of providing ideas for further studies on pharmacological activity of propolis, as well as reference for in-depth development of propolis products.
Animals ; Antioxidants ; chemistry ; isolation & purification ; pharmacology ; Geography ; Humans ; Propolis ; chemistry ; isolation & purification ; pharmacology

The clinical features and lung CT findings of 174 heart failure patients,including 85 cases with normal left ventricular ejection fraction (LVEF) and 89 cases with reduced LVEF were reviewed.Patients with normal LVEF had a higher proportion of hypertension,diabetes and obesity than patients with normal LVEF; and cough and dyspnea were more common.The lung CT findings in patients with normal LVEF were frequently presented as interstitial lung edema,increased pulmonary texture,subpleural line,Kerley lines and diffuse ground-glass opacity.Patients with reduced LVEF had a higher proportion of coronary heart disease and clinical manifestations of acute left heart failure,and central alveolar edema presented in lung CT images.Results suggest that heart failure patients with normal LVEF usually have longer clinical history and occult symptoms,and have a different lung CT imaging features from those in heart failure patients with reduced LVEF.

Objective To investigate the effects of human leptin ( hLEP) gene transfection on rat bone marrow stro-mal cells ( rBMSCs) .Methods rBMSCs were cultured and transfected with adenoviruses encoding hLEP ( Ad5-hLEP-EGFP) in vitro as experimental group while rBMSCs transfected with Ad 5-EGFP and non-transfected were control groups.The proliferation was detected by MTT and the expression of collagen type Ⅰ(Col-Ⅰ) and alkaline phosphatase ( ALP) were assessed by real-time PCR.The ability of mineralized nodule forming was also examined by Alizarin red staining .The combination of transfected rBMSCs and β-tricalcium phosphate (β-TCP ) was con-structed and the osteogenic ability of the construction was evaluated in nude mice .Results hLEP could be trans-fected into rBMSCs successfully by adenovirous .After transfection , the proliferation was not affected while Col-Ⅰand ALP expressions were more pronounced in rBMSCs transfected with Ad 5-hLEP-EGFP ( P<0.05 ) .Alizarin red staining showed the ability of mineralized nodule forming was also up-regulated in Ad5-hLEP-EGFP group (P<0.05).In addition, the transfected rBMSCs adhered to β-TCP and survived well and the combination showed more new bone like tissue formation in nude mice compared to control groups .Conclusions rBMSCs transfected with hLEP might be potently used in bone or periodontal tissue regeneration .

Accidents, Occupational ; prevention & control ; Carbon Dioxide ; poisoning ; Facility Design and Construction ; Humans

Crystallization has been widely applied in pharmaceutical formulations as an effective approach to improve the stability and efficacy of small agents. However protein crystals are suffered from limitation in the drug delivery system due to their complex crystallization behaviors. With development of crystallization technologies and their industrial application, protein crystals are receiving more and more attentions as a novel delivery system for biomacromolecules. Crystals with thermodynamic stable structure can improve the physical and chemical stability of protein drugs and present a sustained release behavior. On the basis of pertinent literatures, this review introduces the recent research situation and development process of protein crystals as drug delivery system. Moreover, the crystallization process of proteins, as well as the preparation and potential application are discussed systematically.
Crystallization ; Drug Delivery Systems ; Pharmaceutical Preparations ; administration & dosage ; Proteins ; chemistry

Objective To establish a homothermal and fast detecting method on pathogenic bacteria by combining recombinase-aid amplification (RAA) with molecular beacon.Methods The establishment of the methodology.Staphylococcus aureus specific primers were designed from the relative region of the staphylococcal protein A (SPA).Asymmetry amplification was optimized by adjusting the primer concentration ratios.The results of amplification and hybridization were visualized and analyzed by agarose gel electrophoresis and fluorescence detection.The sensitivity was identified by detecting dilute positive plasmids.And the specificity was determined using RAA method by detecting 72 pathogenic bacteria,including Staphylococcus aureus and other Staphylococcus spp.from the Department of Clinical Laboratory of Daping Hospital in December 2016.Besides,the Kappa analysis and the clinical diagnosis efficiency were investigated by analyzing 39 extra strains in the laboratory in December 2016.Results When the concentration ratio of restrictive and non-restrictive primer was 1:20,the yield efficiency of single-stranded DNA (ssDNA) reached the peak.And as for the hybridization efficiency,the asymmetry amplification was higher than symmetry amplification.Twenty copies/μl was proposed as the limits of detection by testing dilute plasmids.And the RAA hybridization method could distinguish Staphylococcus aureus with other Staphylococcus spp.Comparing with traditional detection methods with a Kappa index of 0.860,this method shows a good consistency.By analyzing the 111 bacteria,the sensitivity of the method is 92.5％ (37/40),the specificity is 97.2％ (69/71),the positive predictive value is 94.9％ (37/39),the negative predictive value is 95.8％ (69/72),the positive likelihood radio is 33.04,the negative likelihood radio is 0.077,the Youden index is 0.897 and the Kappa index is 0.902.Conclusion Through the combination of asymmetry recombinase-aid amplification optimization and molecular beacon probe,a new method of detecting bacteria DNA with RAA hybridization technique is established,providing the foundation for its clinical application.

Objective:To discuss the treatment options for patients with azoospermia factor (AZF)c microdeletion on Y chromosome.Methods:One hundred and eighty three patients,who were diagnosed as AZFc microdeletion on Y chromosome in Peking University Third Hospital,were recruited in our stu-dy.In order to get better treatment option for this kind of patients,we retrospectively analyzed their clinic data including the treatment process and pregnancy outcome and found out the characteristics of their se-men.Results:Among the 183 patients,sperms can be found in ejaculated semen in 105 patients (57.4%,105 /183).One hundred and three patients (98.1%,103 /105)were diagnosed as severe or extremely severe oligospermia.Regular medication was given to 98 patients,6 patients (6.1%,6 /98) of which got natural pregnancy.The other 99 patients who have sperms in their semen received intracyto-plasmic sperm injection (ICSI),68 patients (68.7%,68 /99)of which got pregnancy.Seventy eight patients were diagnosed as azoospermia among all the 183 patients.Forty nine patients received testicular sperm aspiration (TESA),and 21 patients choose to receive micro-TESE directly.Among the 49 patients with TESA,sperms were retrieved in 17 patients (34.7%,17 /49),and sperms were not retrieved in 32 patients (65.3%,32 /49),of which 12 patients (37.5%,12 /32)gave up treatment and 20 patients (62.5%,20 /32)choose micro-TESE.Among the 41 patients who choose to receive micro-TESE,ope-ration has been done on 19 patients,of which 11 patients (57.9%,11 /19)got sperms.Among the 11 patients,TESA has been done on 6 patients before micro-TESE,of which 4 patients (66.6%,4 /6)got sperms.ICSI has already been done on 7 azoospermia AZFc microdeletion patients who underwent micro-TESE,of which 4 patients (57.1%,4 /7)get pregnancy.Conclusion:AZFc microdeletion patients who had sperms were always diagnosed as severe or extremely severe oligospermia.ICSI was their first choice instead of drug therapy.For AZFc microdeletion patients who were diagnosed as azoospermia, TESA was one of their choices,however,the success rate is not high.Micro-TESE is still possible to get sperms even after the failure of TESA.Therefore,we may choose micro-TESE instead of TESA in some azoospermia patients in order to reduce surgical trauma on patients.

This study was aimed to clone mouse adam10 gene promoter and construct its dual luciferase report vector, and to investigate its transcriptional activity. Total DNA was extracted from mouse brain and used for amplifying the fragment containing adam10 gene promoter by PCR. The amplified product was inserted into pGL-4.10 vector to construct pGL4.10-adam10. The pGL4.10-adam10 and control plasmid pGL4.74 were co-transfected into HEK293 FT cells by lipofectamine 2000. The activity of adam10 gene promoter was assayed by luciferase system. The results showed that the recombinant plasmid pGL4.10-adam10 containing promoter of mouse adam10 was correctly constructed. The method was optimized by changing ratio of two plasmids. Moreover, the transcriptional activity of pGL4.10-adam10 stimulated by ionomycin increased. It is concluded that the dual luciferase reporter system is successfully established, which is useful in bioluminescence imaging technology in vitro. The effect of ionomycin can enhance the transcriptional activity of adam10 gene promoter.
ADAM Proteins ; genetics ; ADAM10 Protein ; Amyloid Precursor Protein Secretases ; genetics ; Animals ; Cell Line ; Cloning, Organism ; Genes, Reporter ; Genetic Vectors ; Luciferases ; genetics ; Membrane Proteins ; genetics ; Mice ; Mice, Inbred BALB C ; Plasmids ; Promoter Regions, Genetic