AIM: To study the neuroprotective roles of neuroglobin (NGB) over-expression in the SH-SY5Y cells transfected with pAPPswe.METHODS:The plasmid pEGFP-NGB was successfully constructed and transfected into the SH-SY5Y cells, which were pretreated with pAPPswe.MTT assay was applied to detect the effect of NGB over-expres-sion on the cell survival rates.JC-1 staining was used to detect the level of mitochondrial transmembrane potential.The cell apoptosis was analyzed by flow cytometry.The effects of NGB over-expression on the protein level of p-Akt, Akt and caspase-3/9 were determined by Western blotting.The generation of Aβ42 in the cells was measured by ELISA.RE-SULTS:The cell survival rate was remarkably increased after transfection with NGB compared with control group and emp-ty plasmid group (P<0.05).The over-expression of NGB significantly inhibited the decrease in mitochondrial membrane potential induced by pAPPswe.The over-expression of NGB inhibited the apoptosis of the cells.Furthermore, over-expres-sion of NGB not only inhibited the expression of caspase-3 and caspase-9, but also induced the production of p-Akt, which was prevented by LY294002, an inhibitor of PI3K/Akt.The generation of Aβ42 was inhibited in the cells with the over-ex-pression of NGB.CONCLUSION: Over-expression of NGB significantly inhibits the SH-SY5Y cell injuries induced by pAPPswe and inhibits the expression of caspase-3/9, which is tightly related with cell apoptosis.Furthermore, the neuro-protective roles of NGB may be via activating PI3K/Akt signaling pathway.

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 explore the international development trends and research hotspots of outdoor activities affecting the progression of children’s myopia, and to provide a reference for researching on effective ways to prevent children’s myopia. Methods: Totally 291 relevant documents included in the "Web of Science" core set database were used as research objects, and CiteSpace software was used for visual analysis. Results: At present, the publications in this field were mainly in the United States(81), China(80), Australia(76), and Singapore(33); the top three research institutions were "Natl Univ Singapore"(29), "Australian Natl Univ"(27), "Capital Med Univ"(25); the main authors were "Saw SM", "Morgan IG", "Mitchell P". The field has been developed on the basis of "Ophthalmology", "Public, Environmental and Occupational Health", and has been integrated into 32 disciplines. The research content included "exploration of high risk factors for the progression of children’s myopia" and "outdoor activities", "intervention in children’s progression of myopia" and "longitudinal tracking of children’s vision development". Randomized clinical trials that longitudinally track the correlation between changes in eyeballs and the progression of myopia and the effects of outdoor activities on the biological characteristics of children’s eyeballs have become a hot topic in this field. Conclusion Research on the effects of outdoor activities on the progression of myopia in children has increased dramatically. The study of increasing outdoor activities to interfere with the progression of myopia in children and the vertical tracking of key factors affecting the biological characteristics of children’s eyeballs have become the current international trends.

Objective:To explore the role of negative pressure drainage in the treatment of duodenal stump fistula after gastrectomy.Methods:The data of 36 patients with duodenal stump fistula admitted in our department from January 2016 to January 2020 were analyzed retrospectively. According to the drainage mode of patients with intestinal fistula, the patients were divided into negative pressure drainage group (observation group) and conventional rubber tube drainage group (control group), 18 cases in each group. The incidence of complications and healing time of intestinal fistula in the two groups were observed.Results:There was no significant difference ( P>0.05) in the incidence of complications between the observation group (5.56%) and the control group (11.11%). There was a significant difference in the mean healing time between the observation group [(12.83±6.25)days] and the control group [(23.33±4.64)days] ( P<0.05). Conclusions:Negative pressure drainage is beneficial to the healing of duodenal stump fistula, and does not improve the incidence of postoperative complications. It is safe and effective. It can be used as a conventional drainage for gastrectomy.

Objective:To study the short-term effect of early application of intestinal microecological therapy after gastric cancer surgery.Methods:A retrospective analysis was made on 96 patients with early and middle stage gastric cancer who underwent surgical treatment in the department of gastrointestinal surgery of Shulan (Hangzhou) Hospital from June 1, 2020 to May 31, 2021. Among them, 48 patients in the observation group were given enteral nutrition support treatment in the early stage after operation and intestinal microecological preparation, while the control group of 48 patients in the early postoperative were given enteral nutrition support. The serum total protein, albumin, prealbumin and other nutrition related indexes and immune related indexes such as CD4 + , CD8 + , CD4 + /CD8 + were detected before and 7 days after operation. The clinical indexes such as the time of first anal exhaust and the incidence of infectious complications were recorded. Results:There was no significant difference between the two groups in preoperative serum total protein, albumin, prealbumin and immune related indexes such as CD4 + , CD8 + , CD4 + /CD8 + ( P>0.05). 7 days after operation, the above indexes in the observation group were significantly higher than those in the control group (all P<0.05). The anal exhaust recovery time of observation group was faster ( P<0.05); There was no significant difference in the incidence of postoperative infectious complications between the two groups ( P>0.05). Conclusions:Early application of intestinal microecological agents in patients with gastric cancer after operation can significantly improve the nutritional status and immune function, promote the recovery of intestinal function, and will not increase the incidence of complications.

Pain in the cervical region of air force pilots, who are exposed to high G-forces, is a specifically occupational health problem. To minimize neck problems, the cervical muscles need specific strength exercise. It is important that the training for the neck must be carried out with optimal resistance in exercises. The model NSE-1 neck training machine for pilots was designed for neck strengthening exercises under safe and effective conditions. In order to realize the functions of changeable velocity and resistant (CVR) training and neck isometric contractive exercises, the techniques of adaptive hydraulics, sensor, optic and auditory biological feedback, and signal processing were applied to this machine. The training system mainly consists of mechanical parts (including the chair of flexion and extension, the chair of right and left lateral flexion, the components of hydraulics and torque transformer, etc.), and the software of signal processing and biological feedback. Eleven volunteers were selected for the experiments of neck isometric contractive exercises, three times a week for 6 weeks, where CVR training (flexion, extension, right, left lateral flexion) one time a week. The increase in relative strength of the neck (flexion, extension, left and right lateral flexion) was 70.8%, 83.7%, 78.6% and 75.2%, respectively after training. Results show that the strength of the neck can be increased safely, effectively and rapidly with NSE-1 neck training machine to perform neck training.
Aerospace Medicine ; Aircraft ; Equipment Design ; Exercise ; physiology ; Humans ; Military Personnel ; Neck Muscles ; injuries ; physiology ; Neck Pain ; etiology ; prevention & control ; Physical Education and Training ; Posture

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 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.

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.