AIM:To observe the effects of neuroglobin ( NGB) overexpression on the apoptosis induced by Aβin the brains of double transgenic AD ( APPswe/PS1dE9) mice and to explore its potential mechanisms .METHODS:Twenty-four 13-month-old double transgenic AD mice were randomly divided into 3 groups:intracerebroventricular injection with normal saline (NS) group, intracerebroventricular injection with pcDNA3.1 and NS group, and intracerebroventricular injection with pcDNA3.1 and pNGB group.Immunohistochemistry was used to detect the expression of Aβ1-42 in the brains. TUNEL staining was used for analyzing the apoptosis , and the protein levels of cleaved caspase-3, caspase-9, PI3K, Akt and p-Akt were determined by Western blot .RESULTS: After intracerebroventricular injection with pNGB , the areas of Aβ1-42 in the hippocampus and cortex were decreased compared with NS group and pcDNA 3.1+NS group (P<0.01).The TUNEL-positive staining cells in the pNGB group were less than those in NS group and pcDNA 3.1 group ( P<0.01 ) . NGB overexpression attenuated the protein levels of cleaved caspase-3 and caspase-9 (P<0.01), but induced the produc-tion of PI3K and p-Akt (P<0.01).CONCLUSION:Overexpression of pNGB significantly inhibits the generation of Aβand attenuates the apoptosis induced by Aβ, indicating that NGB overexpression activates PI 3K/Akt pathway and inhibits the production of cleaved caspase-3 and caspase-9, which were tightly related with apoptosis .

Objective:To observe the stoichiometry of vascular endothelial growth factor receptor 2 (VEGFR2) on the retinal vascular endothelial cell membrane by single-molecule fluorescence imaging.Methods:Rhesus monkey retinal vascular endothelial cells (RF/6A) were divided into blank control group (normal culture) and plasmid transfection group [transfected with VEGFR2-green fluorescent protein (GFP) recombinant plasmid]. The expression of GFP in the plasmid transfected group was observed by confocal microscope, and the expression of VEGFR2 in the cells was detected by real-time fluorescent quantitative polymerase chain reaction (qPCR) and Western blot. The fluorescence intensity distribution and bleaching steps of single VEGFR2-GFP molecule on the cell membrane were recorded by single-molecule imaging. The distribution of fluorescence intensity and the number of fluorescence bleaching steps of GFP were recorded.Results:GFP green fluorescence was observed in the transfected cells 12 hours after transfection. qPCR results showed that the expression of VEGFR2 and GFP mRNA in the plasmid transfected group was significantly higher than that in the blank control group ( t=11.240, 12.330; P<0.001, 0.001). Western blot results showed that the expression of VEGFR2 protein in the plasmid transfected group was significantly higher than that in the blank control group ( t=8.346, P<0.01). The results of single-molecule imaging showed that the fluorescence intensity distribution of VEGFR2-GFP on the surface of RF/6A cell membrane without ligand stimulation was bimodal, in which monomer and dimer were 86.0% and 14.0% respectively. By counting the steps of GFP fluorescence bleaching, the proportions of receptor monomer, dimer, trimer, and tetramer were 81.4%, 12.9%, 5.5%, and 0.3% respectively. Conclusion:In the absence of ligands, VEGFR2 coexists in the form of monomers and dimers on the surface of RF/6A cell membrane, and monomers are dominant.

Objective:To investigate the preoperative and intraoperative risk factors of clinical pancreatic fistula after laparoscopic pancreaticoduodenectomy (LPD).Methods:Clinical data of 100 patients undergoing LPD at the Second Department of Hepatobiliary and Pancreatic Surgery of the First Hospital of Jilin University from Jan 2019 to May 2019 were studied.Results:The total incidence of pancreatic fistula was 8%, and the incidence of clinical pancreatic fistula (grade B, C) was 6%. Univariate analysis found that gender (male), preoperative pancreatic plain CT value <33HU, soft texture of the pancreas, long operation time, alcoholic consuming history may be risk factors for pancreatic fistula after LPD( P<0.05), and further multivariate analysis found gender (male), preoperative pancreatic plain CT value <33 HU, soft texture of the pancreas were independent risk factors for clinical pancreatic fistula after LPD( P<0.05). Conclusions:Male patients, preoperative pancreatic plain CT value <33HU, soft texture of the pancreas were respectively independent risk factors predicting post-LPD clinical pancreatic fistula.

Objective:To investigate the effects of hydroxysafflor yellow A (HSYA) on depressive-like behavior and expression of type A γ-aminobutyric acid receptor(GABAAR)in hippocampus of chronic restraint stress model mice.Methods:The SPF grade male C57BL/6C mice were divided into Control group, HSYA group, Model group, Model + HSYA group and Model + fluoxetine group according to random number table method, with 12 mice in each group.Mice model of depression was established by chronic restraint stress.Mice in HSYA group and Model+ HSYA group were intraperitoneally injected with HSYA(20 mg/kg), mice in Model+ fluoxetine group were injected intraperitoneally with fluoxetine (10 mg/kg), and mice in Control group and Model group administered with 0.9% sodium chloride solution intraperitoneally once a day for 14 days.Then, the forced swimming test (FST) and tail suspension test (TST) were performed to evaluate the depressive-like behavior of mice, and the protein expression levels of different subtypes of GABAAR in the hippocampus of mice were determined by Western blot.SPSS 19.0 and GraphPad Prism 8.0 software were used for data statistical analysis and mapping.One-way ANOVA was used for comparison among groups, and Tukey-HSD test was used for further pairwise comparison.Results:(1) In the behavioral tests, there were significant differences in swimming immobility time of FST and tail suspension immobility time of TST among the five groups ( F=21.59, 20.81, both P<0.05). The swimming immobility time ((143.91±9.97) s) and tail suspension immobility time (( 107.00±6.54) s) in Model group were higher than those in Control group ((52.92±6.70) s, ( 43.50±5.96) s, both P<0.05). There were no significant difference in swimming immobility time and tail suspension immobility time between Model+ HSYA group ((26.17±7.69)s, ( 20.17±7.89)s) and Model+ fluoxetine group ((61.60±16.22)s, (34.14±10.74)s)(both P>0.05), but the swimming immobility time and tail suspension immobility time in these two groups were lower than those in Model group (both P<0.05). (2) The Western blot results showed that there were significant differences in the expression of GABAARβ1 and GABAARβ2 protein in hippocampus among the four groups ( F=12.21, 11.40, both P<0.05). The expression levels of GABAARβ1(45.60±10.76) and GABAARβ2 (46.27±4.82) protein in hippocampus of Model group were lower than those in Control group ((100.00±3.44), (100.00±3.26), both P<0.05). Compared to Model group, the expression of GABAARβ1 (79.91±5.00) and GABAARβ2 (79.08±5.53) protein in hippocampus of Model+ HSYA group were higher (both P<0.05). In addition, the expression of GABAARα1 and GABAARγ1 proteins in hippocampus were not significantly different among the four groups( F=0.23, 0.10, both P>0.05). Conclusion:HSYA can effectively alleviate depressive-like behavior in depression model mice, which may be related with the upregulation of GABAARβ1 and GABAARβ2 of hippocampus tissue.

Objective To compare the differences of neck injury risks for different mummies during arrested landing on naval vessels. Methods Based on the established dynamic model of dummy-belt-seat systems and mechanical model of head-neck, with the acceleration curve at seat base during typical arrested landing reported in the literature as the input, numerical simulation was conducted to test the validity of the model, and the axial force, shear force， bending moment of typical joints in dummy neck during arrested landing were obtained. Results The injury index Nkm of 95 percentile dummy was higher than that of 5 percentile dummy. The injury index Nij of the dummy’s upper and lower cervical spine was smaller than 0.2, meanwhile the Nkm was around 0.65. Conclusions During arrested landing, human neck was much easier to get hurt due to the shear force. The research findings can provide data support for evaluating neck injury risks of pilots with different sizes during typical arrested landing process.

Neovascularization is a characteristic manifestation of a variety of retinal diseases. Vascular endothelial growth factor (VEGF) mainly regulates the proliferation and migration of endothelial cells. VEGF receptor 2 (VEGFR2) is the main receptor to mediate this effect. The activation of downstream signals requires the binding of VEGF and VEGFR2, followed by receptor dimerization and autophosphorylation. Blocking this process and inhibiting neovascularization is very attractive treatment ideas. Monoclonal antibodies and fusion protein drugs currently used in ophthalmology can bind free VEGF. In addition, there are also macromolecular antibodies binding VEGFR2 and small molecule tyrosine kinase inhibitors, which is expected to further expand into the field of ophthalmology. Although anti-VEGFR2 therapy is a revolutionary method to inhibit neovascularization, there are no sufficient clinical evidences at present. In-depth understanding of the application status and progress of anti-VEGFR2 in the treatment of retinal neovascular diseases has important clinical significance.

OBJECTIVE：To establish HPLC fingerprint of Coptis chinensis inflorescence，and study its spectrum-effect relationship with antioxidant and antibacterial effects. METHODS ：Taking 14 batches of C. chinensis inflorescence from different producing areas as the object ，HPLC method was adopted. The determination was performed on Supersil C 18 column with mobile phase consisted of acetonitrile- 0.1％ phosphoric acid solution（gradient elution ）at the flow rate of 1.0 mL/min. The column temperature was set at 25 ℃. The detection wavelength was set at 329 nm，and sample size was 10 μL. The fingerprints of 14 batches of C. chinensis inflorescence were established by Similarity Evaluation System of TCM Fingerprint （2012 A edition ），and the similarity evaluation and common peak identification were carried out. Taking DPPH free radical scavenging rate and hydroxyl radical scavenging rate as antioxidant effects index ，relative antibacterial activity （Escherichia coli ）as antibacterial effect index ， SPSS 21.0 software was adopted to analyze the Pearson correlation between common peaks of C. chinensis inflorescence and above efficacy indexes ；their spectrum-effect relationship was established and validated. RESULTS ：A total of 7 common peaks were obtained in HPLC fingerprint of C. chinensis inflorescence，and the similarity was no less than 0.916. No. 5 peak was identified as berberine hydrochloride. Seven common peaks were positively correlated with DPPH free radical scavenging rate ；No. 1-3，4，6，7 peaks were positively correlated with hydroxyl radical scavenging rate ，while No. 5 peak was negatively correlated with hydroxyl radical scavenging rate. There was a positive correlation between No. 5 peak and antibacterial activity in vitro . After validation ， relative error between the predicted values and the measured values of DPPH free radical scavenging rate ，hydroxyl radical scavenging rate and relative antibacterial activity was 0.92％- 14.5％ . CONLUSIONS ：The established spectrum-effect relationship model can be used to evaluate antioxidant andantibacterial effects of C. chinensis inflorescence. The chemical components represented by No. 1，2，3，4，6，7 peaks are the material basis of antioxidant effect of C. chinensis inflorescence， and berberine hydrochloride is the material basis of antibacterial effect.

Neovascularization is the hallmark of many fundus diseases, including diabetic retinopathy, retinal vein occlusion and neovascular age-related macular degeneration.More and more evidence suggests that vascular endothelial growth factor (VEGF) plays a critical role in neovascularization.Anti-VEGF drugs are the first-line treatment for neovascular fundus diseases and have achieved significant results.However, there are drawbacks such as short drug half-lives and the need for long-term administration to maintain effective concentrations, which increases the economic burden and medical risk for patients and reduces compliance.Therefore, finding a new method for intraocular drug delivery is of great clinical importance.Based on the principle that diabetes patients use insulin pumps to gradually release drugs, the ocular anti-VEGF drug delivery system can continuously release anti-VEGF drugs over a period of time, significantly reducing the injection frequency and improving patient compliance.At present, the research on ocular anti-VEGF drug delivery systems is still immature, and various systems are in different stages of clinical trials.According to different design principles, they can be divided into three categories with their characteristics, micropump (extraocular storage delivery systems), biodegradable implants, and non-biodegradable implants.This article summarized and analyzed the controlled ocular anti-VEGF drug release delivery systems currently in clinical trials.

Abiotic stresses substantially affect the growth and development of plants. Plants have evolved multiple strategies to cope with the environmental stresses, among which transcription factors play an important role in regulating the tolerance to abiotic stresses. Basic leucine zipper transcription factors (bZIP) are one of the largest gene families. The stability and activity of bZIP transcription factors could be regulated by different post-translational modifications (PTMs) in response to various intracellular or extracellular stresses. This paper introduces the structural feature and classification of bZIP transcription factors, followed by summarizing the PTMs of bZIP transcription factors, such as phosphorylation, ubiquitination and small ubiquitin-like modifier (SUMO) modification, in response to abiotic stresses. In addition, future perspectives were prospected, which may facilitate cultivating excellent stress-resistant crop varieties by regulating the PTMs of bZIP transcription factors.
Basic-Leucine Zipper Transcription Factors/genetics* ; Protein Processing, Post-Translational ; Phosphorylation ; Transcription Factors/genetics* ; Stress, Physiological/genetics*