Effect of Long-term Peripheral Nerve Stimulation on Neuro-Muscular Complex Morphologic Recovery in Experiment / Efecto de la Estimulación del Nervio Periférico a Largo Plazo sobre la Recuperación Morfológica del Complejo Neuromuscular en Experimento

SUMMARY: Peripheral nerve injury is an extremely important medical and socio-economic problem. It is far from a solution, despite on rapid development of technologies. To study the effect of long-term electrical stimulation of peripheral nerves, we used a domestically produced electrical stimulation system, which is approved for clinical use. The study was performed on 28 rabbits. Control of regeneration was carried out after 3 month with morphologic techniques. The use of long-term electrostimulation technology leads to an improvement in the results of the recovery of the nerve trunk after an injury, both directly at the site of damage, when stimulation begins in the early period, and indirectly, after the nerve fibers reach the effector muscle.

La lesión de los nervios periféricos es un problema médico y socioeconómico extremadamente importante. Sin embargo, y a pesar del rápido desarrollo de las tecnologías, aún no tiene solución. Para estudiar el efecto de la estimulación eléctrica a largo plazo de los nervios periféricos, utilizamos un sistema de estimulación eléctrica de producción nacional, que está aprobado para uso clínico. El estudio se realizó en 28 conejos. El control de la regeneración se realizó a los 3 meses con técnicas morfológicas. El uso de tecnología de electro estimulación a largo plazo conduce a una mejora en los resultados de la recuperación del tronco nervioso después de una lesión, tanto directamente en el lugar del daño, cuando la estimulación comienza en el período temprano, como indirectamente, después de que las fibras nerviosas alcanzan el músculo efector.

Optimization of ethanol extraction process for Yihuang powder by orthogonal experiment combined with AHP and GA-BP neural network / 中国药房

OBJECTIVE To optimize ethanol extraction process of Yihuang powder. METHODS An orthogonal experiment was designed by reflux extraction with ethanol volume fraction， liquid-to-material ratio， and extraction time as investigation factors. The parameters used were the contents of hesperidin， nobiletin， tangeretin， gallic acid， chebulagic acid， chebulinic acid， liquiritin， glycyrrhizin， eugenol， and the paste-forming rate. The analytic hierarchy process （AHP） was used to calculate the comprehensive score. The optimal ethanol extraction process parameters of Yihuang powder were determined by verifying the results predicted by orthogonal experiment and genetic algorithm （GA）-back propagation neural network （BP neural network）. RESULTS The optimal ethanol extraction process parameters， as optimized by orthogonal experiment， were as follows: ethanol volume fraction of 60%， liquid-solid ratio of 14∶1 （mL/g)， extraction time of 90 min， and extraction for 2 times. The comprehensive score obtained by verification was 79.19. Meanwhile， the optimal ethanol extraction process parameters， optimized by GA-BP neural network， were ethanol volume fraction of 65%， liquid-solid ratio of 14∶1 （mL/g )， extraction time of 60 min， and extraction for 2 times. The comprehensive score obtained by verification was 85.30， higher than the results obtained from orthogonal experiment. CONCLUSIONS The optimization method of orthogonal experiment combined with GA-BP neural network is superior to the traditional orthogonal experiment optimization method. The optimized ethanol extraction process of Yihuang powder is stable and reliable.

Study on preparation technology of long-acting sustained-release oral ulcer membrane based on analytic hierarchy process and orthogonal design / 药学实践杂志

Objective To prepare a sustained-release membrane with longer adhesion time and dissolution time, and compare it with the commercially available oral ulcer membrane. Method Adhesion strength, adhesion time, swelling coefficient, dissolution time, etc. were used as the inspection indicators, and a combination of single factor inspection and analytic hierarchy process were used to screen the membrane -forming materials. The dispersion method of clotrimazole, ornidazole and borneol were investigated to prevent the drug from seed out. The method of combining orthogonal experiment and analytic hierarchy process were used to optimize the dosage of CMC-Na, PVA-1788 and glycerin; and the commercial products were compared. Results Through single-factor investigation and orthogonal experiment, the optimal ratio of excipients was selected as CMC-Na∶PVA-1788∶glycerol (3∶1∶0.08). The water-insoluble component clotrimazole, ornidazole and borneol were treated by precipitation in liquid with good effect. The best method was used to prepare the membrane. The adhesion strength was 102 g. The adhesion time was 55 min. The swelling coefficient was 1 939.52. The average dissolution time was 110 min. The appearance was white and the surface was free of bubbles, soft and elastic. The membrane forming time at 60 ℃ was 300 min and the demolding effect was better which could be completely peeled off with moderate thickness. Conclusion The oral ulcer membrane developed in this method has good appearance, comfortable use, strong adhesion, long adhesion time and dissolution time, and could stay on the ulcer surface for a long time to form physical isolation, and slowly release the drug during the dissolution process, which could play the role of long-term pain relief, antibacterial, anti-inflammatory and promote healing effects on oral ulcers.

Hepatocyte steatosis activates macrophage inflammatory response accelerating atherosclerosis development / 浙江大学学报·医学版

OBJECTIVES@#To investigate the mechanism of comorbidity between non-alcoholic fatty liver disease (NAFLD) and atherosclerosis (AS) based on metabolomics and network pharmacology.@*METHODS@#Six ApoE－/－ mice were fed with a high-fat diet for 16 weeks as a comorbid model of NAFLD and AS (model group). Normal diet was given to 6 wildtype C57BL/6J mice (control group). Serum samples were taken from both groups for a non-targeted metabolomics assay to identify differential metabolites. Network pharmacology was applied to explore the possible mechanistic effects of differential metabolites on AS and NAFLD. An in vitro comorbid cell model was constructed using NCTC1469 cells and RAW264.7 macrophage. Cellular lipid accumulation, cell viability, morphology and function of mitochondria were detected with oil red O staining, CCK-8 assay, transmission electron microscopy and JC-1 staining, respectively.@*RESULTS@#A total of 85 differential metabolites associated with comorbidity of NAFLD and AS were identified. The top 20 differential metabolites were subjected to network pharmacology analysis, which showed that the core targets of differential metabolites related to AS and NAFLD were STAT3, EGFR, MAPK14, PPARG, NFKB1, PTGS2, ESR1, PPARA, PTPN1 and SCD. The Kyoto Encyclopedia of Genes and Genomes showed the top 10 signaling pathways were PPAR signaling pathway, AGE-RAGE signaling pathway in diabetic complications, alcoholic liver disease, prolactin signaling pathway, insulin resistance, TNF signaling pathway, hepatitis B, the relax in signaling pathway, IL-17 signaling pathway and NAFLD. Experimental validation showed that lipid metabolism-related genes PPARG, PPARA, PTPN1, and SCD were significantly changed in hepatocyte models, and steatotic hepatocytes affected the expression of macrophage inflammation-related genes STAT3, NFKB1 and PTGS2; steatotic hepatocytes promoted the formation of foam cells and exacerbated the accumulation of lipids in foam cells; the disrupted morphology, impaired function, and increased reactive oxygen species production were observed in steatotic hepatocyte mitochondria, while the formation of foam cells aggravated mitochondrial damage.@*CONCLUSIONS@#Abnormal lipid metabolism and inflammatory response are distinctive features of comorbid AS and NAFLD. Hepatocyte steatosis causes mitochondrial damage, which leads to mitochondrial dysfunction, increased reactive oxygen species and activation of macrophage inflammatory response, resulting in the acceleration of AS development.

Mechanism of Wuling Capsules against hepatic fibrosis based on network pharmacology and animal experiments / 中国中药杂志

The present study aimed to explore the underlying mechanism of Wuling Capsules in the treatment of hepatic fibrosis(HF) through network pharmacology, molecular docking, and animal experiments. Firstly, the chemical components and targets of Wuling Capsules against HF were searched from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP), Traditional Chinese Medicines Integrated Database(TCMID), GeneCards, and literature retrieval. The protein-protein interaction(PPI) network analysis was carried out on the common targets by STRING database and Cytoscape 3.9.1 software, and the core targets were screened, followed by Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analyses. Enrichment analysis was conducted on the core targets and the "drug-core component-target-pathway-disease" network was further constructed. Subsequently, molecular docking between core components and core targets was conducted using AutoDock Vina software to predict the underlying mechanism of action against HF. Finally, an HF model induced by CCl_4 was constructed in rats, and the general signs and liver tissue morphology were observed. HE and Masson staining were used to analyze the liver tissue sections. The effects of Wuling Capsules on the levels of inflammatory factors, hydroxyproline(HYP) levels, and core targets were analyzed by ELISA, RT-PCR, etc. A total of 445 chemical components of Wuling Capsules were screened, corresponding to 3 882 potential targets, intersecting with 1 240 targets of HF, and 47 core targets such as TNF, IL6, INS, and PIK3CA were screened. GO and KEGG enrichment analysis showed that the core targets mainly affected the process of cell stimulation response and metabolic regulation, involving cancer, PI3K-Akt, MAPK, and other signaling pathways. Molecular docking showed that the core components of Wuling Capsules, such as lucidenic acid K, ganoderic acid B, lucidenic acid N, saikosaponin Q2, and neocryptotanshinone, had high affinities with the core targets, such as TNF, IL6 and PIK3CA. Animal experiments showed that Wuling Capsules could reduce fat vacuole, inflammatory infiltration, and collagen deposition in rat liver, decrease the levels of inflammatory cytokines TNF-α, IL-6, and HYP, and downregulated the expressions of PI3K and Akt mRNA. This study suggests that the anti-HF effect of Wuling Capsules may be achieved by regulating the PI3K-Akt signaling pathway, reducing the levels of TNF-α and IL-6 inflammatory factors, and inhibiting the excessive deposition of collagen.

Mechanism of Shaoyaotang in Treatment of Ulcerative Colitis Based on Network Pharmacology and Experimental Verification / 中国实验方剂学杂志

ObjectiveTo explore the mechanism of Shaoyaotang （SYT） in the treatment of ulcerative colitis （UC） based on network pharmacology and experimental verification. MethodThe core components， target genes， and main pathways of SYT were predicted based on network pharmacology， and UC-related components， target genes， and pathways were screened. Dextran sodium sulfate （DSS） was used to induce the UC model in mice， and the effect of SYT on UC mice was observed， followed by mechanism verification. ResultNetwork pharmacology indicated that 174 active components and corresponding 159 target genes of SYT were screened， and the related pathways were those mediated by 5-hydroxytryptamine （5-HT） degredation and 5-HT receptor 3. The results of animal experiments showed that compared with the model group， the SYT group showed increased body weight and colon length（P<0.01）， reduced disease activity index （DAI） score （P<0.01）， improved histopathological manifestations， reduced concentrations of 5-HT in the colonic tissues and serum （P<0.05， P<0.01）， and increased mRNA expression of monoamine oxidase A （MAOA）， monoamine oxidase B （MAOB）， sodium-dependent serotonin transporter （SLC6A4）， and 5-HT receptor 3A （5-HTR3A） related to 5-HT metabolism in the colon （P<0.01）. ConclusionSYT can alleviate the local inflammatory response of the intestinal tract in UC by regulating 5-HT degredation pathways.

Animal experimental study of 5G remote robot-assisted thoracoscopic lobectomy / 中国胸心血管外科临床杂志

@#Objective     To explore the feasibility of 5G remote robot-assisted pulmonary lobectomy through animal experiments. Methods     In this research, the Toumai® surgical robot was manipulated remotely by the surgeon in the Control Center of the MedBot Company through the 5G network established by China Telecom, and the experimental pig underwent lobectomy in simulated operating room. Results     The animal experiment surgery was successfully completed. The surgeon remotely manipulated the surgical robot to complete the lobectomy of right apical lobe and mediastinal lymph node dissection. The entire animal experiment took about 60 minutes, with an average round-trip network delay of 125 (110-155) ms, and no network interruption or robot malfunction occurred. Conclusion     This animal experiment is the first attempt of 5G remote thoracic surgery, which preliminarily proves the feasibility of completing remote lobectomy through the Toumai® surgical robot 5G wireless network connection. The systematic surgical procedure is summarized, which lays a foundation for the subsequent experiments and clinical applications of 5G remote robot-assisted thoracic surgery.

Preference of nursing staff on providing " Internet + nursing service" for elderly patients / 中华医院管理杂志

Objective:To find the preferences of nursing staff when they provide " Internet + nursing service" for elderly patients.Methods:By means of the discrete choice experiment, a questionnaire was designed to investigate the preferences of nursing staff when they provide " Internet + nursing services" for elderly patients. In-service nursing staff from 8 medical and health institutions in Ningbo city and Wenzhou city were selected by random sampling, for an online questionnaire survey conducted from May to June 2022. The preferences of nursing staff on their service income, service content, service distance, service continuity and residence status of elderly patients when providing " Internet + nursing service" were analyzed by mixed logit regression.Results:A total of 420 valid questionnaires were collected. Compared to 50 yuan/order, nursing staff preferred to a price of 150 yuan/order ( β= 1.22, P<0.001) nursing services; Compared to specialized nursing services, nursing staff preferred to the routine care ( β= 0.86, P<0.001) and health promotion ( β= 0.86, P<0.05) service; Compared to<5 km, nursing staff were unwilling to provide nursing services for elderly patients at distances of 5-10 km and 11-15 km ( β=-0.66, P<0.05; β=-0.95, P<0.001) ; Compared to 1-2 visits per month, nursing staff preferred not to provide continuing care services ( β=-0.70, P<0.05); Compared to homestay with the patient family, nursing staff preferred to provide nursing services for elderly patients residing in nursing homes ( β= 1.21, P<0.001) . Conclusions:Considering the preference of nursing staff tend to provide " Internet + nursing service" for the elderly patients with services featuring appropriate price, non-specialist care, close distance, low continuity (moderate intensity used as the reference) and security assurance for practice.

Metformin inhibiting the activation of NLRP3 inflammasome and pyroptosis in diabetic retinal vascular endothelial cells / 中华眼底病杂志

Objective:To observe the effect of metformin (Met) on inflammatory bodies and focal death in human retinal microvascular endothelial cells (hRMEC) in diabetes mellitus (DM) microenvironment.Methods:Experimental research was divided into in vivo animal experiment and in vitro cell experiment. In vivo animal experiments: 9 healthy C57BL/6J male mice were randomly divided into DM group, normal control group, and DM+Met group, with 3 mice in each group. DM group and DM+Met group mice were induced by streptozotocin to establish DM model, and DM+Met group was given Met 400 mg/ (kg · d) intervention. Eight weeks after modeling, the expression of NLRP3, cleaved-membrane perforating protein D (GSDMD) and cleaved-Caspase-1 in the retina of mice in the normal control group, DM group and DM+Met group were observed by immunohistochemical staining. In vitro cell experiments: hRMEC was divided into conventional culture cell group (N group), advanced glycation end products (AGE) group, and AGE+Met group. Joining the AGE, AGE+Met groups cells were induced by 150 μg/ml of glycation end products, and 2.0 mmol/L Met was added to the AGE+Met group. Pyroptosis was detected by flow cytometry; 2' ,7'-dichlorofluorescein diacetate (DCFH-DA) fluorescent probe was used to detect the expression of reactive oxygen species (ROS) in cells of each group. Real-time fluorescence quantitative polymerase chain reaction and Western blot were used to detect the relative mRNA and protein expression levels of NLRP3, cleaved-GSDMD, cleaved-Caspase-1 in each group of cells. Single factor analysis of variance was used for comparison among the three groups.Results:In vivo animal experiments: compared with the DM group, the expression of NLRP3, cleaved-GSDMD, and cleaved-Caspase-1 in the retina of normal control group and DM+Met group mice was significantly reduced, with significant difference among the 3 groups ( F=43.478, 36.643, 24.464; P<0.01). In vitro cell experiment and flow cytometry showed that the pyroptosis rate of AGE group was significantly higher than that of N group and AGE+Met group ( F=32.598, P<0.01). The DCFH-DA detection results showed that the intracellular ROS levels in the N group and AGE+Met group were significantly lower than those in the AGE group, with the significant difference ( F=47.267, P<0.01). The mRNA ( F=51.563, 32.192, 44.473; P<0.01) and protein levels ( F=63.372, 54.463, 48.412; P<0.01) of NLRP3, cleaved-GSDMD, and cleaved-Caspase-1 in hRMEC of the AGE+Met group were significantly reduced compared to the N group. Conclusion:Met can down regulate the expression of NLRP3 inflammatory body related factors in hRMEC and inhibit pyroptosis.

Construction of virtuality and reality combination teaching mode of molecular biology experiment based on constructivism / 中华医学教育探索杂志

Objective:Molecular biology experimental technology has become an important basic tool for exploring biology and medicine and other related disciplines. We aim to explore an effective molecular biology experimental teaching model which will definitely improve students' molecular biology experimental skills and autonomous learning ability.Methods:Guided by the theory of constructivism, with the molecular biology experimental course as the carrier, and with the basic requirements of constructing the basic molecular biology experimental technology of the system, a teaching platform was established to guide students to preview the experiment independently; the physical experimental projects were integrated and optimized and the virtual simulation experimental projects were increased, with virtuality and reality, to improve students' molecular biology experimental skills and autonomous learning ability.Results:An online teaching platform has been established, which effectively guides and improves the effect of students' preview experiments, and cultivates the ability of autonomous learning. Besides, the experimental teaching mode combining optimization of physical experimental projects and virtual simulation experimental projects significantly improved students' molecular biology experimental operation skills and problem-solving ability.Conclusion:A constructivism-based teaching mode of combining virtual and real molecular biology has been established, which is an effective way to improve students' molecular biology experimental skills and autonomous learning ability.

Ethical problems and solutions of experimental animals in functional experiment teaching / 中华医学教育探索杂志

Objective:To investigate the ethics and existing problems of experimental animals in medical experiments and experimental teaching, and to formulate countermeasures so that animal ethics and animal welfare can be truly reflected in medical experiments.Methods:In this study, a "Basic Function Experiment Center Animal Experiment Questionnaire" with 25 questions was formulated from three aspects: the ethical cognition of experimental animals, whether animal experiments are ethical or not, how to view the problems of animal ethics and experimental teaching and the cognition of virtual simulation experiment teaching. Questionnaire was sent to Hubei University of Medicine to investigate the international students of Batch 2017 (5-year program), undergraduates of Batch 2017 (5-year program) and nursing students of Batch 2018 (4-year program) as well as teachers, researchers and employees of laboratory animal center (all with bachelor degree or above). The survey results were expressed as percentage.Results:The recovery rate of the questionnaire in this study was 98.04%(2 451/2 500), among which the practitioners, teachers and researchers in the laboratory animal center clearly understood the ethics of experimental animals, but there was a widespread phenomenon of lagging ethics among the students. For example, 16.24% (398/2 451) students had not received animal experiment ethics education and training, 29.46% (722/2 451) were not clear about animal protection laws and regulations, 7.14% (175/2 451) thought animal experiments were immoral; 29.54% (724/2 451) had vague cognition of animal welfare and ethical knowledge; 25.91% (635/2 451) were not familiar with the operation steps; 9.38% (230/2 451) were indifferent to the extra treatment of animals due to operation errors, 7.83% (192/2 451) chose to give up the experiment in the treatment of animals after massive bleeding, only 5.43% (133/2 451) chose to continue the experiment after timely hemostasis and infusion, and 9.26% (227/2 451) chose to do operations unrelated to the experiment. After the experiment, 2.28% (56/2 451) chose to kill the animals by bloodletting, only 5.51% (135/2 451) chose excessive anesthesia euthanasia, and 1.96% (48/2 451) chose to kill the animals by cervical dislocation and violence. Only 15.79% (387/2 451) chose to remember the dead animal for 2 minutes. Only 32.56%(798/2 451) of the respondents understood virtual simulation experiment, 34.92% (856/2 451) of the respondents thought that virtual simulation experiment or experimental teaching video could be used to replace the existing live animal experiment, 77.56% (1 901/2 451) believed that the construction of virtual simulation laboratory should be strengthened.Conclusion:It is imperative to strengthen the education of students' ethics of experimental animals, which is conducive to the establishment of correct ethics of experimental animals for medical students, so that the "3R" principle and animal welfare can be truly implemented in experimental teaching and scientific research experiments.

Study on the construction and teaching exploration for virtual simulation experiment of whole process of infant nutritional status evaluation / 中华医学教育探索杂志

Infant nutritional status evaluation is a basic experimental skill in the experimental teaching of nutrition and food hygiene. The construction of this virtual simulation experiment can solve the problems of activeness and poor coordination of infants and young children in the actual operation of the experiment. And based on the application of this virtual simulation project, the experimental teaching method of preventive medicine was explored. This experiment adopted online and offline mixed teaching method, which enriches teaching measures and strengthens the standardized process of infant nutritional status evaluation. After 3 semesters of practice from spring 2018 to spring 2020, the proportion of students who achieved excellent grades was 30.09% (34/113), 56.02% (279/498) and 66.79% (1 080/1 617), respectively, which increased significantly year by year ( Ptrend < 0.001). Among all the 2 228 students, 1 983 students (89%) believed that this experimental teaching could better cultivate the ability of autonomous learning. Through the study of virtual simulation experiments, students have improved their subjective initiative, and laid a foundation for the improvement of students' overall quality and the requirements of school elite education.

Exploration and thinking of online teaching of medical microbiology experiment / 中华医学教育探索杂志

Medical microbiology experiment is faced with many problems in online teaching. This study adopts the teaching mode of online live broadcast + operation video + virtual experiment, and make up the operation gap to some extent through operation video and virtual experiment. The mode of assessment is subjective thinking question (closely following the operation process) + experiment design + literature review (focusing on the key technology or new technology of clinical assessment that cannot be carried out due to the limitation of conditions in traditional experiments, such as mass spectrometry, fluorescence quantitative PCR, and G-test), and it is helpful to understand students' mastery of teaching objectives, and the ability of comprehensive application and innovative thinking. The student questionnaire shows that most students hold a positive attitude towards the online experimental teaching mode, and the quality of students' homework shows that most students have a good learning effect.

Application of "Flipped Classroom" in comprehensive experiment teaching of Digital Signal Processing for biomedical engineering specialty / 中华医学教育探索杂志

"Flipped Classroom" is a new kind of "student-centered" teaching model, which can give full play to the advantages of both sides of teaching and learning. According to this teaching model, we redesigned the teaching process, in which the students studied by themselves and built their own knowledge system. Moreover, each of them took part in three stages of experimental design including digital signal collection, analysis and processing in groups. Results have shown that this model can fully stimulate students' learning interest, not only helps students to deepen understanding of digital signal processing theory knowledge, but also strengthen the ability of autonomous learning and team collaboration. The teaching model maybe have certain reference function in comprehensive experiment teaching of Digital Signal Processing course for biomedical engineering specialty.

Evaluation of teaching effect of opening experiments on occupational health and occupational medicine / 中华医学教育探索杂志

Objective:To carry out opening experiments on occupational health and occupational medicine for students majoring in preventive medicine, reform the experimental teaching mode, and explore the teaching methods to improve students' professional quality and scientific research ability.Methods:Opening experiments were carried out in the experimental course of occupational health and occupational medicine for students of preventive medicine major. A total of 147 students majoring in preventive medicine of Batch 2016 were classified as the control group, and the routine confirmatory experiment was carried out in the group; 176 students majoring in preventive medicine of Batch 2017 were classified as the experimental group, and this group carried out opening experiment. The evaluation was made from three aspects: comprehensive evaluation results, teacher self-evaluation and student satisfaction survey. SPSS 22.0 software was used for analysis and comparison by independent-samples t-test and Chi-square test. Results:The theoretical scores of the experimental group and the control group students were (84.37±10.45) vs. (81.44±9.22) ( t=2.68, P=0.008), and the experimental skills scores were (93.66±3.89) vs. (88.41±5.67) ( t=9.51, P<0.001). Questionnaire investigation showed that the students in the opening experimental group were more satisfied with the courses arrangement ( χ2=8.31, P=0.004), group cooperation ( χ2=21.10, P<0.001), assessment form ( χ2 =7.92, P=0.005), improvement of the writing ability of scientific research papers ( χ2 =17.56, P<0.001), improvement of practical skills ( χ2=11.70, P=0.001), logical thinking, language organization and expression ability ( χ2=10.33, P=0.001). They considered the opening experiment was helpful to cultivate innovative thinking and ability, but it had limited effect on the cultivation of employment advantages. And the students considered the opening experiments of setting up professional courses was sufficient and necessary. Conclusion:Carrying out opening experiments for students majoring in preventive medicine is helpful to improve students' professional quality and cultivate their practical ability and scientific research ability.

Search for potential target genes in dexamethasone-induced open-angle glaucoma by bioinformatics / 中华实验眼科杂志

Objective:To predict potential target genes in dexamethasone-induced open-angle glaucoma via bioinformatics technology.Methods:The GEO datasets GSE16643, GSE37474, and GSE124114 were used to analyze the differentially expressed genes by GEO2R.Gene Set Enrichment Analysis (GSEA) was performed on the differentially expressed genes between GSE37474 and GSE124114.Intersection of the three datasets were displayed by Venn diagram.The annotation and enrichment analysis of the intersection genes were performed through Gene Ontology (GO)/Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis and then were compared with normal tissue in GTEx Portal database.The corresponding protein interaction network was obtained by STRING.Finally, the candidate genes were searched for their transcription factors in UCSC and JASPAR.Primary human trabecular cells were divided into dexamethasone group and control group, treated with 2 ml 500 nmol/L dexamethasone and the same amount of ethanol, respectively.The expression of BDKRB1 and TAGLN in trabecular cells was detected by Western blot.Results:Differential genes between GSE37474 and GSE124114 datasets enriched in complement and coagulation cascade by GSEA.There were 89 intersecting genes of the three datasets.These genes mainly regulated the formation of extracellular matrix by GO analysis.The gene with the highest enrichment score and collagen-containing extracellular matrix was found to be associated with fibroblasts in GTEx Portal database.ACTA2, MYL9, TAGLN, and LMOD1 were closely related in STRING protein-protein interaction network.Transcription factor SP1 in UCSC and JASPAR according to related genes, BDKRB1, NID1, MFGE8 and TAGLN.The relative expression levels of BDKRB1 and TAGLN proteins were 1.32±0.14 and 0.44±0.09 in dexamethasone group, respectively, which were significantly higher than 1.00±0.00 and 0.20±0.10 in the control group, respectively ( t=-3.61, 2.89; both at P<0.05). Conclusions:Bioinformatics analysis showed that transcription factor SP1 may play a role in human trabecular meshwork cells to myofibroblasts transition after dexamethasone treatment.

Study on prediction of potential targets and mechanism of Zeqi Decoction in the treatment of lung adenocarcinoma / 国际中医中药杂志

Objective:To explore the potential molecular targets and mechanism of Zeqi Decoction in the treatment of lung adenocarcinoma (LUAD) through bioinformatics and cell experiment.Methods:The active components of Zeqi Decoction were collected based on TCMSP database and literature search. Then R software was used to screen differentially expressed genes in LUAD from TCGA and GEO databases. The co-expression module was obtained through weighted gene co-expression network analysis (WGCNA), and the potential targets were obtained after matching and mapping with targets of Zeqi Decoction. Enrichment analysis of GO function and KEGG pathway of targets was conducted. The results were experimentally verified. The lung adenocarcinoma cell lines A549 and H1299 were divided into blank control group and Zeqi Decoction group according to random number table. The inhibition rate of cell proliferation was detected by cell proliferation test (CCK-8); the expression of leukocyte differentiation antigen 36 (CD36) in A549 and H1299 cells was detected by Western blot; the levels of low density lipoprotein receptor (LDLR) and IL-6 were detected by ELISA.Results:Totally 157 anti-lung adenocarcinoma active components and 18 potential targets were obtained, mainly including CD36, IL6, LDLR, etc. The main target of Zeqi Decoction in the treatment of lung adenocarcinoma was lipid metabolism. The results showed that Zeqi Decoction could effectively inhibit the activity of A549 and H1299 cells and the levels of CD36, LDLR and IL-6.Conclusion:Zeqi Decoction can inhibit the inflammatory response by down-regulating the protein expressions of CD36 and LDLR, thereby slowing the proliferation of cells.

Animal Experiments on Male Infertility Treated by Tonifying Kidney / 中国实验方剂学杂志

Experimental research on male infertility is critical to the study of the pathogenesis of male infertility and the evaluation of drug therapy. This paper reviewed animal experiments on male infertility in recent years. The experimental models of male infertility mainly include oligoasthenozoospermia （OA），teratozoospermia，azoospermia， and varicocele animal models. The OA animal models are mostly induced by glycosides of Tripterygium wilfordii （GTW）， adenine，hydrocortisone， and radiation，which are mainly chemical means. The animal models of azoospermia were usually constructed by intraperitoneal injection of bissulfonyl alkylating agent busulfan and immersion of scrotum in 43 ℃ water. There are few studies on animal models of teratozoospermia，and the induction methods by GTW and methyl methanesulfonate（MMS） are common. The animal models of varicocele-caused infertility are usually induced by operation. The ligation of the middle division of the left renal vein between the lateral inferior vena cava and the medial spermatic vein has a significant influence on testicular morphology and epididymal sperm quality. Animal experimental studies have shown that classic prescriptions for tonifying the kidney and promoting spermatogenesis represented by Wuzi Yanzongwan and clinical empirical prescriptions by modern research have played a significant role in the treatment of male infertility. The mechanism of tonifying the kidney in the treatment of male infertility mainly focuses on inhibiting spermatogenic cell apoptosis. The kidney-tonifying method can regulate the apoptosis of spermatogenic cells，which provides a new treatment idea and a reliable scientific basis for traditional Chinese medicine in the field of male reproduction.

Application of Design of Experiment in development of purification process of recombinant human growth hormone-Fc fusion protein / 中国生物制品学杂志

@#ObjectiveTo optimize the condition for anion exchange chromatography in purification process of recombinant human growth hormone-Fc(rhGH-Fc)fusion protein by Design of Experiment(DOE)so as to decrease the content of host cell protein(HCP)in bulk of protein.MethodsA complete factorial experimental design with four factors at two levels was established by the DOE in Minitab 19 software.The condition(flow rate,sample load,pH value of buffer and salt concentration of eluent)for anion exchange chromatography in purification process of rhGH-Fc was optimized by DOE to find out the operating space.ResultsThe experimental results were predicted accurately by the established DOE model.The sample load,pH value of buffer,salt concentration of eluent as well as the interaction of pH value of buffer and salt concentration of eluent showed significant influence on the HCP content in the harvest.The optimal sample load,flow rate as well as pH value and salt concentration of eluent were 15 mg/mL,120 cm/h,7.0 ～ 8.0 and 0.1 mol/L respectively.The HCP contents in eluents under the optimal condition were less than 400 ng/mg,which met the requirements for verification within the range of results in determined operating space.ConclusionThe condition for removal of HCP by anion exchange chromatography was successfully optimized by DOE,which provided a reference for further application of DOE in the biopharmaceutical field.

Exploration and practice of innovative teaching of molecular biology experiment course under the context of new agricultural science / 生物工程学报

The development of new agricultural science is a new idea and a new measure that aims to deepen the reform of higher education in agriculture and forestry for the development of new agriculture, new countryside, new farmers and new ecosystem. It is therefore essential for the current undergraduate experimental teaching reform to timely introduce new technologies and new methods used in the development of agriculture and forestry industry into the experimental course teaching, and promote the integration of professional education with innovation and entrepreneurship education, according to the requirements of the new agricultural science. In view of this, the exploration and practice of molecular biology experiment course was carried out from the perspective of experimental projects, teaching modes and evaluation methods, according to the teaching requirements and characteristics of molecular biology experiment and teachers' scientific research achievements. The results showed that this reform greatly improved the students' comprehensive quality and innovation ability and may facilitate the innovation experiment teaching of other courses.