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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.
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Animals , Rabbits , Electric Stimulation/methods , Peripheral Nerve Injuries/therapy , Peripheral Nerves , Muscle, Skeletal/innervation , Recovery of Function , Nerve RegenerationABSTRACT
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.
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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.
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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.
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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.
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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.
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"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.
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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.
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【Objective】 To design and simulate routine serological experiments in transfusion techniques using RhD blood group, so as to solve the problem of difficult to obtain positive specimens in experimental teaching. 【Methods】 RhD positive red blood cells, RhD negative red blood cells and anti-D reagent were used to design and simulate the enzyme treatment experiment, absorption and elution test, antibody identification experiment and cross matching experiment of polybrene technology in transfusion techniques. 【Results】 Papain treatment of red blood cells made the agglutination of RhD positive red blood cells and IgG anti-D visible. Absorption and elution test were successfully simulated with RhD positive red blood cells and IgG anti-D reagent. The antibody identification of anti-Jka and anti-Fya was successfully simulated by creating different identification panel and panel cells made by RhD positive cells and negative cells. Cross matching test of polybrene method can also be simulated using RhD negative and positive red blood cells and IgG anti-D reagent. 【Conclusion】 RhD blood group can be used to simulate most of the routine serological experiments of blood transfusion, which can be used in the serological laboratory teaching of transfusion.
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【Objective】 To discuss the regulating effect of stored red blood cells (RBCs) transfusion on BMDMs in inflammatory conditions, and the relationship between stored RBCs transfusion and inflammatory response induced by bacterial infection. 【Methods】 Forty C57BL/6 male mice of 6-8 weeks (18-22 g/mouse) were randomly divided into experimental group and control group. Each mouse was infected with 200 µL Pseudomonas aeruginosa injecting into the tail vein, and 400 µL fresh (storage >14 d) and stored RBCs (storage 0.05). F4/80 of experimental group and control group 2, 4 and 8 hours after RBCs infused were 1.83±0.11 vs 0.75±0.06, 0.46±0.06 vs 0.33±0.06 (P0.05), respectively. iNOS, TNF-α, MCP1 of M1 in liver of experimental group and control group 2, 4 and 8 hours after RBCs infused were respectively: iNOS 3.44±0.20 vs 2.46±0.08, 9.25±0.55 vs 2.67±0.12, 2.80±0.08 vs 2.39 ±0.01; TNF-α 1.69±0.22 vs 1.13±0.03, 1.44±0.24 vs 0.96±0.09, 1.31±0.05 vs 0.96±0.06; MCP1 4.96±0.08 vs 4.28±0.27, 4.63±0.04 vs 2.07±0.09, 2.28±0.19 vs 1.33±0.03 (P0.05). 【Conclusion】 Stored RBCs infusion can greatly promote the M1 polarization of BMDMs in liver.
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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.
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@#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.
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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.
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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.
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OBJECTIVE To explore the intestinal absorption characteristics of saikosaponins. METHODS Based on everted intestinal sac model， using accumulative absorption amount （Q） and absorption rate constant （Ka） as indexes， UHPLC-MS/MS technique as a method， the absorption of saikosaponin A， B2， C， D and F from total saponins of Bupleurum chinense （8 g/mL， by crude drug） in the duodenum， jejunum and ileum was detected. RESULTS The correlation coefficients （r） of the regression equations for the absorption of saikosaponins A， B2， C and F in the duodenum， jejunum and ileum were all higher than 0.95， while the r of saikosaponin D in the above intestinal segments was lower than 0.95； compared with the absorption of the same composition in the duodenum， the Q and Ka of saikosaponin A and C circulating in jejunum and ileum for 120 min， as well as the Q and Ka of saikosaponin F circulating in the ileum for 120 min were significantly decreased （P＜0.05）. CONCLUSIONS Saikosaponin A and the other 4 saikosaponins are all absorbed in the duodenum， jejunum and ileum； among them， saikosaponin A， B2， C and F are linearly absorbed， which conforms to the zero-order absorption characteristics， but saikosaponin D shows non- linear absorption.
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In 2015, the United States put forward the concept of precision medicine, which changed medical treatment from "one size fits all" to personalization, and paid more attention to personalization and drug customization. In the same year, Spritam®, the world's first 3D printed tablet, was in the market, marking the emerging pharmaceutical 3D printing technology was recognized by regulatory authorities, and it also provided a new way for drug customization. 3D printing technology has strong interdisciplinary and high flexibility, which puts forward higher requirements for pharmaceutical staffs. With the development of artificial intelligence (AI), modern society can perform various tasks, such as disease diagnosis and robotic surgery, with superhuman speed and intelligence. As a major AI technology, machine learning (ML) has been widely used in many aspects of 3D printing drug, accelerating the research and development, production, and clinical application, and promoting the new process of global personalized medicine and industry 4.0. This paper introduces the basic concepts and main classifications of 3D printing drug, non-AI drug optimization technology and ML. It focuses on the analysis of the research progress of ML in 3D printing drug, and elucidates how AI can empower the intelligent level of 3D printing drug in pre-processing, printing, and post-processing process. It provides a new idea for accelerating the development of 3D printed drug.
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Cerebral hemorrhage accounts for about 10%-15% of all strokes， and its pathogenesis is complex. Currently， the main clinical treatment is mainly medical symptomatic treatment， including the use of antihypertensive drugs， hypoglycemic drugs， and hemostatic drugs， and surgical treatment is required in some cases， but there is still a lack of effective treatment. In recent years， traditional Chinese medicine and proprietary Chinese medicine have been widely accepted for their stable efficacy， high safety， and low cost. Rhei Radix et Rhizoma is one of the most commonly used herbal medicines for the treatment of cerebral hemorrhage. This paper summarizes the relevant literature on the treatment of cerebral hemorrhage with Rhei Radix et Rhizoma and finds that its active ingredients are mainly anthraquinones， such as emodin， Rhei Radix et Rhizoma acid， and Rhei Radix et Rhizoma phenol. The herbal formulas are Da Chengqitang， Shengdi Dahuangtang， Liangxue Tongyufang， and Naoxueshu oral liquid. The effects involve protecting the blood-brain barrier， promoting hematoma absorption， reducing inflammation levels， decreasing lactic acid accumulation at the bleeding site， and increasing the expression of brain-derived neurotrophic factors. The pathways involved include aquaporin 4 （AQP4）， phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt）， extracellular signal-regulated kinase 1/2 （ERK1/2）， Toll-like receptor 4 （TLR4）， nuclear transcription factor-κB （NF-κB）， nuclear factor E2-related factor 2 （Nrf2）， and Wnt3a/β-linked protein pathway. This paper summarizes the progress of clinical studies and animal experiments on the treatment of cerebral hemorrhage with active ingredients of Rhei Radix et Rhizoma and herbal compounds containing Rhei Radix et Rhizoma， so as to provide a reference for the treatment protocol of cerebral hemorrhage.
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@#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.
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ObjectiveTo investigate the effect of Ganoderma lucidum polysaccharides （GLP） on the proliferation， migration， cycle， and apoptosis of hepatocellular carcinoma SKHEP1 and Huh7 cells and to explore the underlying mechanism. MethodSK-HEP-1 and Huh-7 cells were classified into the blank group and low-， medium-， and high-dose GLP groups （3.5， 7， 14 g·L-1）. The proliferation of the cells was examined by cell counting kit-8 （CCK8） assay， and the migration by scratch assay. Cell cycle was measured by flow cytometry and apoptosis was detected based on Hoechst33258 staining. In addition， the expression of phosphatidylinositol 3-kinase （PI3K）， protein kinase B （Akt）， phosphorylated PI3K （pPI3K）， and phosphorylated Akt （pAkt） in the cells was determined by Western blot. ResultCompared with the blank group， the three doses of GLP reduced the proliferation and migration of SKHEP1 and Huh7 cells （P<0.05）， increased the percentage of cells in G1 phase （P<0.05）， and decreased percentage of cells in S and G2 phase （P<0.05）. In addition， the three doses can induce apoptosis of both SK-HEP-1 and Huh-7 cells， particularly the high dose. Moreover， the three doses of GLP lowered the levels of pPI3K and pAkt （P<0.05）. ConclusionGLP significantly inhibited the malignant phenotype of SK-HEP-1 and Huh-7 cells through the PI3K/Akt signaling pathway.
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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.