ObjectiveTo explore the effects of astragalin （AST） on autophagy and apoptosis of astrocytes in the L_4-5 dorsal horn of the spinal cord in mice with inflammatory pain induced by complete Freund's adjuvant （CFA）. MethodsTwenty-four male C57BL/6 mice， aged six months， were randomly assigned to four groups： control group， saline group， CFA model group， and CFA+AST group， six mice in each group. The inflammatory pain model was established by injection of 10 µL CFA into the right lateral malleolus fossa. The saline group were injected with an equal amount of normal saline at the same site. The inflammatory pain mice in CFA+AST group were further treated with AST （60 mg/kg） intraperitoneally once a day for 21 consecutive days. Multiplex immunofluorescence staining was used to detect the coexpression of autophagy-related factors including ATG 12 and Beclin-1， apoptosis-related factors including Cleaved-Caspase3 and Caspase9， and the astrocyte marker such as GFAP in the L_4-5 spinal dorsal horn of the mice in each group. Western blot was used to examine the protein expression levels of autophagy-related proteins（ATG12， Beclin-1） and apoptosis-related proteins（Caspase 3， Caspase 9） in the L_4-5 spinal dorsal horn of mice. ResultsImmunofluorescent staining showed that in the L_4-5 dorsal horn of the spinal cord， the fluorescence intensity of ATG12 （P<0.000 1） and Beclin-1 （P<0.000 1） was significantly increased， while that of Cleaved-Caspase 3 （P<0.001） and Caspase 9 （P<0.000 1） was decreased in the CFA+AST group when compared to the CFA model group. Furthermore， AST could inhibit the activation of astrocytes. Western blot further confirmed that AST significantly upregulated the expression of ATG12 （P<0.000 1） and Beclin-1 （P<0.000 1） in the L_4-5 spinal cord of CFA mice， and downregulated the expression of Caspase 3 （P<0.01） and Caspase 9 （P<0.001）. ConclusionsAST promotes autophagy of astrocytes and inhibits their apoptosis in the L_4-5 spinal dorsal horn of CFA mice.

Geobacillus thermoglucosidasius is a kind of Gram-positive facultative anaerobic bacteria. The fast growth rate under high temperature and less susceptibility to microbial contamination enable G. thermoglucosidasius to be a desirable producer of biofuels and high-value-added chemicals for the next-generation industrial biotechnology. However, compared with the classical model strain Escherichia coli, the applications of G. thermoglucosidasius are hampered by its low transformation efficiency. This study aimed at obtaining competent cells with high transformation efficiency through inactivating restriction enzymes, adding cell membrane inhibitors and cell wall weakening agents. The results showed that the electro-transformation efficiency achieved 1.2×10⁴ CFU/(μg DNA) by knocking out four genes encoding restriction enzymes. Adding a certain amount of tween 80, dl-threonine and glycine further increased the competent efficiency about 22.5, 44, and 334 times, respectively. The electro-transformation efficiency was enhanced to 4.6×10⁶ CFU/(μg DNA) under the optimized conditions, laying a foundation for genetic manipulation and metabolic engineering of G. thermoglucosidasius.
Electroporation ; Electroporation Therapies ; Bacillaceae ; Cell Membrane ; Escherichia coli/genetics*

Objective:To establish an artificial intelligence (AI)-assisted diagnostic system for lung cancer via deep transfer learning.Methods:The researchers collected 519 lung pathologic slides from 2016 to 2019, covering various lung tissues, including normal tissues, adenocarcinoma, squamous cell carcinoma and small cell carcinoma, from the Beijing Chest Hospital, the Capital Medical University. The slides were digitized by scanner, and 316 slides were used as training set and 203 as the internal test set. The researchers labeled all the training slides by pathologists and establish a semantic segmentation model based on DeepLab v3 with ResNet-50 to detect lung cancers at the pixel level. To perform transfer learning, the researchers utilized the gastric cancer detection model to initialize the deep neural network parameters. The lung cancer detection convolutional neural network was further trained by fine-tuning of the labeled data. The deep learning model was tested by 203 slides in the internal test set and 1 081 slides obtained from TCIA database, named as the external test set.Results:The model trained with transfer learning showed substantial accuracy advantage against the one trained from scratch for the internal test set [area under curve (AUC) 0.988 vs. 0.971, Kappa 0.852 vs. 0.832]. For the external test set, the transferred model achieved an AUC of 0.968 and Kappa of 0.828, indicating superior generalization ability. By studying the predictions made by the model, the researchers obtained deeper understandings of the deep learning model.Conclusions:The lung cancer histopathological diagnostic system achieves higher accuracy and superior generalization ability. With the development of histopathological AI, the transfer learning can effectively train diagnosis models and shorten the learning period, and improve the model performance.

Objective To establish the GIOP model and extract BMSCs from the rat model.We aim to in-vesitigatethe effect ofrhPTH(1-34)for inhibiting β-catenin ubiquitination when combining with Micro-CT and bio-logical technology.We also investigate the influence of rhPTH(1-34)on the GIOP.Methods Female SPF emale rats wererandomly divided into normal control group,methylprednisolone group(model group),methylpredniso-lone+saline group(blankcontrol group)and methylprednisolone+rhPTH(1-34)group(test group). The proximal femoral cancellous bone was examined by Micro-CTand histopathological Staining. The expression of Wnt10b and β-catenin protein were detected. By comparing with inducedBMP-2,BMSCs were treated withrhPTH(1-34)and stained with ALP and alizarin red.Results(1)In Micro-CT,BV/TV,Tb.Th and Tb/N decreased,whereas Tb/sp increased in the test group comparedwith model group(P<0.05).ROI three-dimensional reconstruction of trabecu-lar bone in test group showed local bone repair;(2)Wnt10b and β-cateninexpression increased in the test group compared with the model model(P<0.05),indicating that rhPTH(1-34)can enhance the transcriptional activity of β-catenin(P<0.05)and promote the expression of Wnt10b andβ-catenin(P<0.05).Conclusion The inter-vention with rhPTH(1-34)can prevent GIOP by regulating the Wnt/β-catenin signaling pathway and inhibiting GIOP progress,which can improve the microstructure of bone.

Objective To investigate the distribution characteristics of pathogenic bacteria among different seasons and ages in children lower respiratory tract infection.Methods To retrospectively analyze the results of bacterial culture in pediatric outpatients and inpatients with lower respiratory tract infections in our hospital from September 2014 to August 2015.The pathogenic bacterial detection situation of lower respiratory tract infection among different age groups and different were statistically analyzed.Results A total of 2 809 pathogenic bacterial strains were isolated from 4 629 lower respiratory tract specimens,and detection rate was 60.7%.The pathogenic bacteria distribution was different among different age groups.Haemophilus influenzae (33.3%) ranked the first place in the lower respiratory infection among children aged from 19 d to 7 year old.The detection rates of Haemophilus parainfluenzae,Moraxella catarrhalis and staphylococcus aureus had obvious seasonality,while Streptococcus pneumoniae had no obvious seasonality.Conclusion Pathogenic bacteria causing lower respiratory tract infections in children are different from those in adults,which are affected by age and seasonal change.

Bacterial two-hybrid system is a newly developed method for studying protein-protein interactions. However, in our studies of the interaction of regulatory proteins in Streptomyces, it was found that the bacterial two-hybrid system is not sensitive enough by the blue-and-white selection on X-gal plate. To overcome this drawback, the reason of false positive clone was firstly determined, which was the disturbance of other direct or indirect regulation on lacZ promoter. Then the disturbance was diluted by introducing multicopy lacZ promoter, which drive another reporter gene gfp. By such design, the sensitivity of the modified bacterial two-hybrid system was significantly inproved and the two different reporters also help to decrease the rate of the false positive clones. Further the evaluation of the modifiedd bacterial two-hybrid system indicated that the sensitivity was significantly improved.
Bacteria ; Genes, Reporter ; Promoter Regions, Genetic ; Protein Interaction Mapping ; methods ; Two-Hybrid System Techniques

Purpose To compare the relationship between the maximum standardized uptake value (SUVmax) of PET/CT imaging and tumor markers in lung cancer, and to provide evidence for the comprehensive assessment of non small cell lung cancer. Materials and Methods The results of PET/CT imaging, serum carcinoembryonic antigen (CEA), cytokeratin-19 fragment antigen 21-1 (CYFRA21-1) and neuronspecific enolase (NSE) of 179 patients with non small cell lung cancer were analyzed, subjects were then divided into different groups according to their clinical features and pathological types, the differences of SUVmax and tumor markers'' level were compared between different groups, and their correlations with tumor histological type, differentiation and the clinical stages were also analyzed.Results SUVmax of the tumor was related with lesion size,≥3 cm tumors'' SUVmax were significantly higher than that of <3 cm tumors (P<0.01), SUVmax and serum NSE level were related with tumor differentiation, and these in low differentiation group were higher than those in high and medium differentiation group (P<0.05,P<0.01). In addition, SUVmax, CEA, CYFRA21-1 and NSE had no correlation with gender, age, pathological type and clinical stage (P>0.05).Conclusion SUVmax and NSE have a certain relationship with tumor differentiation in non small cell lung cancer patients, thus can be used as a reference index for prognosis of indirect prediction of lung cancer.

Gene expression exhibits temporal and spatial patterns to response environmental changes and growth cycle. Gene expression is under strict control at different levels among which control at transcription level is the predominant mode, especially in prokaryotes. In this review, we summarized the new developments of methods used in transcriptional studies, including modifications and improvements of the classic methods, such as gel-shift assay, DNA foot printing, and in vivo reporter system. In addition, we introduced examples to apply new methods, such as surface plasmon resonance (SPR) and isothermal titration calorimetry (ITC) to characterize protein-DNA, ligand-protein, and ligand-protein-DNA interactions. The collection of these methods and their application could guide and accelerate relevant studies.
Calorimetry ; DNA Footprinting ; Gene Expression ; Ligands ; Proteins ; Surface Plasmon Resonance ; Transcription, Genetic

BACKGROUND:Clinical studies have demonstrated that systemic mild hypothermia could significantly reduce the disability and mortality rate of patients with severe traumatic brain injury. In recent years, the researches about the treatment of spinal cord injury by mild hypothermia have been successively carried out. OBJECTIVE:To investigate the effects of mild hypothermia on nerve regeneration microenvironment after spinal cord injury and explore the possible underlying mechanism of nerve regeneration and functional recovery after spinal cord injury in rats. METHODS:Twenty out of sixty-seven rats were randomly selected as the sham group, and other rats were used to establish spinal cord injury models in T9 segment using modified Alen's method. Three rats were excluded for failure in spinal cord injury induction and four rats for death during modeling. The rest 40 rats were randomly and evenly divided into the spinal cord injury and mild hypothermia groups (n=20 rats/group). The rats in the sham and spinal cord injury groups were placed in the operating table with normal temperature, making their rectal temperature at 37.0±0.5℃ for 72 hours. The rats in the mild hypothermia group were placed on ice blanket machine, making their rectal temperature at 34.0±0.5℃ for 72 hours, then the temperature was naturaly rewarmed. RESULTS AND CONCLUSION:Compared with spinal cord injury group, the apoptosis index and the level of aquaporin 4/9 mRNA and protein expression in spinal cord injury tissue were al decreased, somatosensory evoked potential latency and amplitude were recovered, and the motor functional scores were increased in the mild hypothermia group. These results indicate that mild hypothermia play its protective effect on spinal cord injury through attenuating apoptosis of neural cels and decreasing aquaporin 4/9 mRNA and protein expression levels.