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BACKGROUND:Pulmonary pericytes are located at the concavity where pulmonary vessels are interconnected,which is closely related to the formation and stability of pulmonary vascularization.However,there are few studies on how pulmonary pericytes affect the activity of pulmonary vascular endothelial cells in the pathogenesis of broncho-pulmonary dysplasia. OBJECTIVE:To analyze the relationship between the quantity of subgroups of pulmonary pericytes and endothelial cells in different stages of broncho-pulmonary dysplasia and to explore the effects of PDGFR-β+NG2+α-SMA+ pericytes on the early tube-forming activity of pulmonary vascular endothelial cells. METHODS:(1)Animal experiment:Twelve newborn C57BL/6 mice were randomly divided into normoxia and hyperoxia groups within 24 hours of birth,with six mice in each group.Mice in the hyperoxia group were exposed to an 85%O2 environment to build the mouse models of broncho-pulmonary dysplasia,while those in the normoxia group were fed in the same room air.The lung tissues of the mice in the two groups were taken at 7 and 14 days after birth.The pathological changes of the lung tissues were observed by hematoxylin-eosin staining.Three subgroups of pulmonary pericytes were measured by flow cytometry:PDGFR-β+NG2+α-SMA-,PDGFR-β+NG2+α-SMA+,and PDGFR-β+NG2-α-SMA+ cells.(2)Cellular experiment:Passage 3 PDGFR-β+NG2+α-SMA+ pericytes were co-cultured with mouse pulmonary vascular endothelial cells(experimental group)at a ratio of 1:4.Mouse pulmonary vascular endothelial cells cultured alone were used as controls.The tube-forming difference between two groups was analyzed after 15 hours of co-culture. RESULT AND CONCLUSION:(1)Animal experiment:Hematoxylin-eosin staining revealed that on day 7,the lung tissue of mice in the normoxia group had regular structure,obvious alveolar structure,and uniform size,while the number of alveoli in the lung tissue of mice in the hyperoxia group was less and the morphology of alveoli was irregular.On day 14,the alveoli of mice in the normoxia group gradually developed and matured,the alveolar structure gradually became regular and uniform in size,and the alveolar density gradually increased.The lung tissue structure of mice in the hyperoxia group was relatively disordered and the alveolar formation was delayed with the size gradually increasing and the alveolar structure being simplified.Flow cytometry results indicated that the number of PDGFR-β+NG2-α-SMA+ and PDGFR-β+NG2+α-SMA+ pericytes was increased in the hypoxia group compared with the normoxia group(P＜0.01),while the number of PDGFR-β+NG2+α-SMA-pericytes and pulmonary vascular endothelial cells was decreased(P＜0.01,P＜0.04).(2)Cellular experiment:In the control group,the pulmonary vascular endothelial cells arranged in cords and extended around,and lumen-like structures formed in some areas.In the experimental group,PDGFR-β+NG2+α-SMA+ pericytes and their pseudopodia were not observed,the irregular grid structure of pulmonary vascular endothelial cells was significantly less than that of the control group,and the endothelial cells mainly clustered in clumps.To conclude,α-SMA+ pericyte subgroups are predominant in mice with broncho-pulmonary dysplasia.PDGFR-β+NG2+α-SMA+ pericytes can directly inhibit the tube-forming activity of pulmonary vascular endothelial cells,which may be involved in the process of abnormal vascularization in broncho-pulmonary dysplasia.
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BACKGROUND:Previous studies have shown that the modified Tsuge suture method can be used to repair chicken tendon injuries.However,the lack of post-repair functional exercise leads to obvious tendon adhesions.Therefore,a functional exercise after tendon repair is very important. OBJECTIVE:To explore a scheme for passive functional exercise against tendon adhesion in a chicken model of flexor toe tendon rupture following repair using the modified Tsuge suture method. METHODS:A total of 100 Sanhuang chickens,10 months of age,were taken to make animal models of deep flexor tendon Ⅱ rupture of the third toe of the right foot.Animal models were randomized into five groups(n=20 per group):groups A,B,C and D were given plaster immobilization for 3 weeks after surgery,and were simultaneously given passive functional exercise 1,2,3,and 0 times a day for 3 weeks,respectively;group E had neither plaster immobilization nor passive functional exercise after surgery.The gross morphology of the chicken claw,the morphology of the tendon anastomosis end and the degree of peritendinous adhesion were observed.The slipping distance of the deep flexor tendon of the third toe,the flexion angles of all joints,the pathological morphology of the tendon at the anastomotic end and the hydroxyproline content were measured. RESULTS AND CONCLUSION:The tendon of group E was completely ruptured around 6 day after surgery and was removed from the experiment.The chicken claws in groups B and C had better grasping morphology,group D had almost no grasping morphology,and group A had worse grasping morphology than groups B and C.In groups B and C,the tendon anastomosis end was not obviously expanded,with the texture similar to that of normal tendon tissue.Compared with group C,tendon adhesion was relatively mild in group B.In group D,the tendon anastomosis end was obviously expanded,with the hard texture,obvious peritendinous scar and serious adhesion.Expansion at the tendon anastomosis end was more obvious than groups B and C but less severe than group D.The slipping distance and the related flexion angles of the deep flexor tendon of the third toe were better in groups B and C than groups A and D(P<0.05).The content of hydroxyproline in the tendon of chickens in group B was higher than those in groups A,C and D(P<0.05).Findings from hematoxylin-eosin and Sirius red staining showed that collagen fibers of the tendon in groups A,C and B were gradually arranged in a directional manner,where the number of bright red and thick type Ⅰ collagen fibers was gradually increased and the number of tiny green type Ⅲ collagen fibers was gradually decreased.In group E,collagen fibers of the tendon were poorly arranged in a directional manner and type Ⅰ and type Ⅲ collagen fibers were cross-distributed.To conclude,adequate passive functional exercises twice a day following repair with the modified Tsuge suture method could effectively alleviate tendon adhesion and reduce tendon rupture in the chicken model of deep flexor tendon rupture.
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BACKGROUND:Mesenchymal stem cells are pluripotent stromal cells isolated from a variety of tissues,which can differentiate into osteoblasts under certain conditions.Photobiomodulation,as an external stimulus,can promote osteogenic differentiation combined with other inducers or alone,providing new ideas for solving a series of bone diseases. OBJECTIVE:To review the relevant literature and mechanisms of photobiomodulation-induced osteogenic differentiation of mesenchymal stem cells,which will lay a theoretical foundation for bone tissue engineering using mesenchymal stem cells as seed cells and may offer some suggestions for future studies. METHODS:Relevant articles were searched on CNKI,PubMed and Wed of Science databases with Chinese search terms of"photobiomodulation,low power laser,low level laser,light-emitting diode,mesenchymal stem cells,osteogenic differentiation,biomaterials"and English search terms of"photobiomodulation,low level laser(light),light-emitting diode(LED),mesenchymal stem cell,osteogenic differentiation,biomaterials".Finally,88 articles were included for analysis. RESULTS AND CONCLUSION:(1)Photobiomodulation represented by low level laser and diode laser has a positive effect on promoting the proliferation and differentiation of mesenchymal stem cells.(2)Photobiomodulation can induce osteogenic differentiation of mesenchymal stem cells,whose feasibility has been verified in cell and animal experiments.On one hand,photobiomodulation can promote the expansion and differentiation of stem cells in vitro by activating related signaling pathways and up-regulating the expression of osteogenic molecules.On the other hand,photobiomodulation can improve the survival rate of stem cells in vivo,promote homing effect and shorten the healing time of bone defects after stem cells are injected into the body.However,photobiomodulation has a biphasic dose effect,whose laser parameters,experimental environment,cell type and other factors in various studies are different,making the research results lack consistency and difficult to apply in the clinic.(3)Combined with biological materials,other physical factors and drugs,photobiomodulation can also accelerate osteogenic differentiation.(4)In conclusion,photobiomodulation has been used increasingly widely in the medical field with its advantages of non-invasive,efficient and less-side reactions,and its role in bone tissue engineering has gradually become prominent,which provides a new method for the treatment of bone defects and related diseases.Further exploration should be focused on the standardized treatment parameters of photobiomodulation.
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BACKGROUND:Overweight or obesity is the most significant risk factors for knee osteoarthritis.Time-restricted diet shows an effective effect in preventing overweight or obesity.Whether infrapatellar fat pad,an important component of the knee joint,is affected by time-restricted diet and thus influences knee osteoarthritis remains unclear. OBJECTIVE:To employ a high-fat diet-induced rat model to investigate the effect of weight loss by time-restricted diet on infrapatellar fat pad,thereby providing evidence for early prevention and treatment of obesity-related knee osteoarthritis. METHODS:Fifteen male Sprague-Dawley rats were randomly divided into three groups(n=5 per group).Rats in the control group were allowed to eat at will for 24 hours and were fed the defined control diet(12%fat);rats in the high-fat diet group were allowed to eat at will for 24 hours and were fed the high-fat diet(45%fat);and rats in the high-fat diet with time-restricted diet group were fed the high-fat diet only from 9:00(2 hours after the light)to 17:00(2 hours before the dark).After 8 weeks of feeding,mDixon-Quant sequence was used to assess proton density fat fraction in the infrapatellar fat pad and subcutaneous adipose tissue in the right inguinal region.ELISA was used to quantify differences adipokine.Sirius red staining was used to evaluate changes in fibrosis of the infrapatellar fat pad.The expressions of uncoupling protein-1 in the infrapatellar fat pad and subcutaneous adipose tissue in the right inguinal region as well as leptin,adiponectin and tumor necrosis factor-α in the infrapatellar fat pad were detected by immunohistochemistry staining. RESULTS AND CONCLUSION:After 8 weeks of feeding,compared with the high-fat diet group,the body mass of rats in the control group(P=0.036)and the high-fat diet with time-restricted diet group(P=0.003)was significantly reduced.The proton density fat fraction in the infrapatellar fat pad in the high-fat diet group was significantly higher than that in the control group(P<0.001)and the high-fat diet with time-restricted diet group(P=0.004),while there was no significant difference in the proton density fat fraction of the subcutaneous adipose tissue among the three groups.The serum leptin levels of rats in the high-fat diet group were significantly higher than those in the control group(P=0.030)and the high-fat diet with time-restricted diet group(P=0.018).Compared with the high-fat diet group,the infrapatellar fat pad fibrosis characterized by Sirius red staining in the control group(P<0.001)and the time-restricted diet group(P=0.003)was significantly decreased.The expression of leptin in IFP of the high-fat diet group was significantly higher than that of the control group(P<0.001)and the high-fat diet with time-restricted diet group(P<0.001).The expression of adiponectin in the infrapatellar fat pad of the high-fat diet group was significantly lower than that of the control group(P=0.004)and the high-fat diet with time-restricted feeding group(P=0.048).However,there was no positive expression of uncoupling protein-1 in the infrapatellar fat pad and subcutaneous adipose tissue of all the three groups,and no positive expression of tumor necrosis factor-α in the infrapatellar fat pad.To conclude,time-restricted diet could retard the fibrosis of the infrapatellar fat pad,reduce the proton density fat fraction of the infrapatellar fat pad,and affect the level of adipokine in serum and infrapatellar fat pad.Time-restricted diet may become a simple and effective option for the treatment and prevention of obesity-related knee osteoarthritis.
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Objective:To compare the feasibility and safety of a new portable endoscopic system and the conventional endoscopic system for the detection and emergency treatment of abdominal trauma in animal models.Methods:Three healthy Bama pigs, which were fasted and water deprivation for 8 h before surgery and then underwent induction anesthesia. A layer-by-layer incision was made into the abdominal cavity of Bama pigs. An artificial pneumoperitoneum was established using a laparoscopic pneumoperitoneum machine. A bullet model was inserted into the abdominal cavity to build the bullet wound model. After the bullet model was removed, a shrapnel model was inserted into the mid-abdomen to build the shrapnel wound model. The two types of endoscopic system were used to detect, remove bullet model or shrapnel model of the three Bama pigs respectively. The procedure order of the two systems was assigned according to the random number table method. The surgical success, operation time, endoscopy pipeline patency, endoscopic operation satisfaction, adverse events and equipment defects were recorded.Results:Three surgeries were performed using the new portable endoscopic system and three other surgeries using the conventional endoscopic system, all of which were successful. The time of the new portable endoscopic system to find and remove the bullet model, and the shrapnel model were 232.33±11.68 s, 300.33±57.70 s, 170.00±44.44 s and 52.67±2.52 s, respectively. The corresponding time of the conventional endoscopic system were 232.67±21.20 s ( t=-0.054, P=0.962), 256.67±67.00 s ( t=0.880, P=0.472), 176.00±52.42 s ( t=-0.111, P=0.922), 58.67±14.84 s ( t=-0.832, P=0.493), respectively. There was no significant difference between the two systems ( P>0.05). The endoscopy tubes of the two endoscopic systems were both smooth. The operator was satisfied with the endoscopic procedures of both endoscopic systems, and no adverse event or device defect occurred. Conclusion:The portable endoscopic system proves to be safe and feasible for the diagnosis and treatment of abdominal trauma in animal models.
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Improving the reproducibility of biomedical research results is a major challenge. Transparent and accurate reporting of the research process enables readers to evaluate the reliability of the research results and further explore the experiment by repeating it or building upon its findings. The ARRIVE 2.0 guidelines, released in 2019 by the UK National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs), provide a checklist that is applicable to any in vivo animal research report. These guidelines aim to improve the standardization of experimental design, implementation, and reporting, as well as enhance the reliability, repeatability, and clinical translation of animal experimental results. The use of the ARRIVE 2.0 guidelines not only enriches the details of animal experimental research reports, ensuring that information on animal experimental results is fully evaluated and utilized, but also enables readers to understand the content expressed by the author accurately and clearly, promoting the transparency and completeness of the fundamental research review process. At present, the ARRIVE 2.0 guidelines have been widely adopted by international biomedical journals. This article is based on the best practices following the ARRIVE 2.0 guidelines in international journals, and it interprets, explains, and elaborates in Chinese the fifth part of the comprehensive version of the ARRIVE 2.0 guidelines published in PLoS Biology in 2020 (the original text can be found at https://arriveguidelines.org). This section includes the items 6-11 of Recommended 11 section, covering "Animal Care and Monitoring", "Interpretation/Scientific Implications", "Generalisability/Translation", "Protocol Registration", "Data Access" and "Declaration of Interests". Its aim is to promote a comprehensive understanding and use of the ARRIVE 2.0 guidelines among domestic researchers, to enhance the standardization of experimental animal research and reporting, and to promote high-quality development of experimental animal sciences and comparative medicine research in China.
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【Objective】 To explore the safety of transrectal ultrasound-guided transperineal injection of sodium hyaluronate to expand the Dirichlet gap in laparoscopic radical prostatectomy. 【Methods】 A total of 14 healthy male purebred beagle dogs were selected and randomly divided into 2 groups, with 7 in either group.The control group was treated with conventional laparoscopic radical prostatectomy, while the experimental group was treated with laparoscopic radical prostatectomy after 2.5 mL sodium hyaluronate was injected into the Dirichlet gap under the guidance of transrectal ultrasound.The total operation time, prostate separation time, intraoperative blood loss and rectal status of the 2 groups were observed. 【Results】 After the injection of sodium hyaluronate into the Dirichlet gap between the prostate and the rectum, no rectal tissue was found in the prostate, and no obvious damage was found in the posterior rectum in either groups.The postoperative hemoglobin (HGB) was [(118.70±2.56) g/L vs.(122.10±2.19) g/L, P=0.02]; the total operation time was [(141.40±9.80) min vs.(119.10±9.16) min, P<0.05]; the prostate separation time was [(24.99±1.75) min vs.(16.64±2.34) min, P<0.05]; the amount of bleeding was [(47.43±4.32) mL vs.(34.86±5.18) mL, P<0.05] in the control group and experimental group. 【Conclusion】 Laparoscopic radical prostatectomy performed after 2.5 mL of sodium hyaluronate injection into the Dirichlet gap under the guidance of transrectal ultrasound can shorten the total operation time, the separation and resection time of the prostate, and reduce the amount of bleeding, which can improve and reduce the incidence of rectal injury, and prove the feasibility of this approach for prostatic cancer.
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Objective:To study the clinical safety and validity of retrograde new endoscopic field of vision in miniature pigs.Methods:6 live miniature pigs were selected as study subjects,En-doscopic Retrograde New View(ERNV)was selected.The performance,image quality and intraoper-ative and postoperative complications were evaluated.To evaluate whether all the experimental ani-mals could complete the relevant endoscopy.Verify ERNV's operating performance,including whether the duodenoscope can enter the biliary tract smoothly,and made sure whether the injection,suction,and instrument channels were unobstructed.Choledochoscope image clarity,color resolu-tion,image deformation and distortion,accurate evaluation of lumen conditions and clear observation of mucosal surface conditions were analyzed.Whether there were operant injuries such as bleeding and perforation,as well as adverse events such as respiratory depression and cardiac arrest.The sur-vival status and adverse reactions of all pigs were observed.Results:The choledochoscope was successfully inserted into the bile duct of 6 miniature pigs.The product had good operation perfor-mance and could enter the bile duct through the duodenoscope smoothly.The injection,suction and instrument channels were relatively smooth.In addition,the endoscopic images are clear,with better color resolution,and without image deformation and distortion,which can realize accurate evaluation of the conditions in the lumen and observe the mucosal surface conditions more clearly.No bile duct stenosis or dilatation occurred in all miniature pigs,and the bile duct mucosa was smooth,without hyperemia and edema,and no abnormal thickening or bending of mucous vessels.During the exami-nation,there were no operational injuries such as bleeding and perforation,and no adverse events such as respiratory depression and cardiac arrest occurred.The vital signs of all miniature pigs tended to be stable after operation,and the survival state was good,and there were no complications such as cholangitis,bleeding and perforation.Conclusion:ERNV has good clinical safety and efficacy,ex-cellent operation performance and excellent image quality,and is worthy of clinical application.
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Despite the record speed of developing vaccines and therapeutics against the SARS-CoV-2 virus, it is not a given that such success can be secured in future pandemics. In addition, COVID-19 vaccination and application of therapeutics remain low in developing countries. Rapid and low cost mass production of antiviral IgY antibodies could be an attractive alternative or complementary option for vaccine and therapeutic development. In this article, we rapidly produced SARS-CoV-2 antigens, immunized hens and purified IgY antibodies in 2 months after the SARS-CoV-2 gene sequence became public. We further demonstrated that the IgY antibodies competitively block RBD binding to ACE2, neutralize authentic SARS-CoV-2 virus and effectively protect hamsters from SARS-CoV-2 challenge by preventing weight loss and lung pathology, representing the first comprehensive study with IgY antibodies. The process of mass production can be easily implemented in most developing countries and hence could become a new vital option in our toolbox for combating viral pandemics. This study could stimulate further studies, optimization and potential applications of IgY antibodies as therapeutics and prophylactics for human and animals.
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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 study the feasibility of domestic single-port surgical robot assisted endoscopic system for partial nephrectomy,and analyze its safety in clinical partial nephrectomy based on experimental results Methods Three qualified experimental pigs were selected,two senior urological professors and a senior resident doctor used a domestic single-port surgical robot to perform partial nephrectomy on the left and right kidneys.Recorded the operation duration,hot ischemia duration,suture time,estimated blood loss,volume of renal parenchyma excision and other information.Results There were 8 wedge resection and 4 heminephrectomies.The kidney volume of wedge resection was(7.35±0.81)mL and the blood loss was(8.50±11.09)mL.The total operation time was(41.67±8.50)min,and the time of resection was(5.88±3.27)min and the stitching time was(11.75±2.82)min.The kidney volume of heminephrectomy was(24.30±2.18)mL,and the blood loss was(6.25±4.35)mL.The total operation time of heminephrectomy was(47.00±11.27)min,and the time of resection was(3.25±1.5)min and the stitching time was(10.00±5.25)min.No bleeding was observed on the wound after the Bull dog was released in all operations.There was no significant difference in operation time and blood loss between the heminephrectomy group and the wedge resection group.There was no significant difference in operation time or blood loss between the senior doctor group and the senior resident doctor group.The NASA-TLX scale was used to assess the degree of workload of the operator during surgical operations,and the results showed that none of the three surgeons had a high level of frustration.There were no adverse events related to the single-port surgical robot system during the operation.Conclusion It is safe and feasible for a domestic single-port surgical robot system to perform a partial nephrectomy.
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Objective:To study the differential expression profiling of the transcripts modified by m5C methylation in a rat model of N-methyl-D-aspartate (NMDA)-induced retinal excitotoxicity.Methods:A total of 65 Sprague Dawley male rats aged 7-8 weeks were randomly divided into two groups: normal control group and NMDA group. The right eye (model eye) of rats in the NMDA group were injected with 50.0 mmol/L of NMDA 3 μl in the vitreous cavity, while in the normal control group, equal volume of normal saline was injected into the vitreous cavity. After 1 week of the injection, the optic nerve conduction function of rats was detected by visual evoked potential. The whole structure of rat retina was observed by hematoxylin-eosin staining, and the thickness of each retinal layer and the number of retinal ganglion cell layer were detected. The number of β3 tubulin immunofluorescence positive cells was detected by immunofluorescence staining on retinal stretched preparation. Total RNA was extracted from the retinas of normal control group and NMDA group, and high-throughput m5C modified RNA was sequenced, and bioinformatics analysis was performed. The relative expression levels of SLFN3, PLXNB3, CD36 and HIC2 mRNA in retina were detected by real-time quantitative polymerase chain reaction. The comparison between the two groups was performed using an unpaired t test. Results:The P1 latency of control group and NMDA group were (117.86±6.48) and (148.46±3.78) ms, and the amplitudes were (42.57±2.41) and (8.68±0.63) μV, respectively. Compared with the normal control group, the latency period was prolonged and the amplitude was significantly decreased in the NMDA group, with statistical significance ( P<0.001). In normal control group, retinal ganglion cells (RGC) were uniformly arranged with large round nuclei. In NMDA group, the volume of retinal RGC was atrophied and the number of RGC was reduced. The total retinal thickness in the control group and NMDA group was (207.51±12.76) μm and (187.51±12.54) μm, respectively. The number of β3 tubulin positive cells was 79.86±6.56 and 29.36±2.16, respectively. Compared with normal control group, the total retinal thickness and the number of β3 tubulin positive cells in NMDA group were decreased, with statistical significance ( P <0.001). Compared with the control group, 576 differentially expressed m5C mRNA were screened in the NMDA group, among which 230 up-regulated and 346 down-regulated genes were detected, respectively. The results of biological information analysis showed that compared with the control group, the upregulated m5C mRNA in the NMDA group was mainly involved in biological processes such as perception and cell-cell adhesion, and was mainly concentrated in the cytokine-cytokine receptor interaction and neural active ligand-receptor interaction pathway. The biological processes in which down-regulated m5C mRNA was mainly involved in biological processes such as G-protein-coupled receptor signaling pathway and cell communication, which were mainly concentrated in primary immune deficiency pathway and neural active ligand-receptor interaction pathway. Real-time quantitative polymerase chain reaction detection results showed that compared with the normal control group, the relative expression levels of SLFN3 and PLXNB3 mRNA in the retina of rats in NMDA group were significantly increased, while the relative expression levels of CD36 and HIC2 mRNA were significantly decreased, with statistical significance ( P<0.05). Conclusion:In NMDA induced retinal excitatory toxicity rat models, m5C modified retinal transcriptome showed abnormal expression.
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Objective:To explore the safety and effectiveness of medium-frequency electrotherapy for increasing the volume of the latissimus dorsi muscle.Methods:Fifteen adult New Zealand white rabbits were randomly divided into three groups, namely group A, group B, and group C, with 5 rabbits in each group. This was a self-control study, with the right latissimus dorsi muscle as the experimental group and the left latissimus dorsi muscle as the control group. The three groups corresponded to three different current intensity levels: 7.062 mA for group A (6th gear), 10.593 mA for group B (9th gear), and 14.124 mA for group C (12th gear). After the 12th, 24th, and 36th sessions of the experiment, ultrasonography was used to collect the thickness of the latissimus dorsi muscle. After the 36th electrostimulation, the latissimus dorsi muscle samples were collected to measure their in vivo muscle thickness and wet weight and were then sent for HE and MASSON staining.Results:After the 12th, 24th, and 36th electrostimulation sessions, ultrasonographic sampling in groups A and B showed an increase in the thickness of the right latissimus dorsi muscle compared to the left; for example, the thickness on the right of group B increased by 37.8%. The wet weight data collected after the 36th electrostimulation in groups A and B showed an increase in the right latissimus dorsi muscle compared to the left; for example, the wet weight on the right of group B increased by 5.04%.Conclusions:Different electrostimulation modes of medium-frequency therapy technology can induce muscle fiber thickening or atrophy. In this experiment, the 9th gear (10.593 mA) of medium-frequency therapy technology may be a suitable choice for inducing muscle fiber thickening, and the 12th gear (14.124 mA) may be a suitable choice for inducing skeletal muscle thinning.
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Objective:To investigate the synergistic antitumor effect of pyrotinib in combination with 5-fluorouracil(5-Fu)on human epidermal growth factor receptor 2(HER2)positive breast cancer cells and its underlying molecular mechanism. Methods:HER2 positive breast cancer cells were screened by Western blotting.HER2 positive SKBR-3 and BT474 cells were treated with pyrotinib and 5-Fu individually or in combination for the following experiments.MTT assay was used to assess the effect of different drugs on the proliferation of the treated cells,and the combination index(CI)values were calculated using Combidrug software.Colony formation assay was used to evaluate the effect of different drugs on the colony-forming ability of the treated cells.FCM assay was used to analyze the effect of different drugs on the apoptosis rate and cell cycle of the treated cells.Western blotting was used to examine the effect of different drugs on the expression levels of proteins in the proliferation-and apoptosis-related signaling pathways.SKBR-3-cell-based tumor xenograft model was established using BALB/c nude mice.After treatment with pyrotinib and 5-Fu individually or in combination,the growth profiles of the xenograft tumors were recorded and the expression levels of proteins in the proliferation-and apoptosis-related signaling pathways were examined in the tumor tissues. Results:HER2 positive breast cancer cell lines SKBR-3 and BT474 were selected for further experiments after screening.The proliferation SKBR-3 and BT474 cells could be inhibited after treatment with pyrotinib and 5-Fu individually or in combination(all P＜0.05).Compared with pyrotinib or 5-Fu single drug treatment,pyrotinib in combination with 5-Fu had higher inhibition rate on the proliferation of SKBR-3 and BT474 cells with a Cl value of＜1,indicating the synergistic effect of pyrotinib and 5-Fu.In addition,in contrast to pyrotinib or 5-Fu single drug treatment,there was a further decrease in the number of colonies formed,increase in apoptosis rate,and increase in the percentage of G0/G,cells in SKBR-3 and BT474 cells after treatment with pyrotinib in combination with 5-Fu(all P＜0.01).Animal experiment results showed that the growth rate of xenograft tumors in mice treated with pyrotinib in combination with 5-Fu was significantly slower than that of the single-drug treated mice(P＜0.05).Western blotting analysis showed that the expression levels of HER2,HER4,AKT and phosphorylated ERK were significantly decreased after treatment with pyrotinib in combination with 5-Fu both in vitro and in vivo(all P＜0.01),indicative of the blockage of proliferation-related signaling pathways.Meanwhile,analysis of the apoptosis-related proteins revealed a decrease in the expression levels of caspase 3,poly ADP-ribose polymerase(PARP),and Bcl-2(all P＜0.01),while an increase in the expression levels of cleaved-caspase 3,cleaved-PARP,and p21(all P＜0.01). Conclusion:Pyrotinib and 5-Fu had synergistical antitumor effect on HER2 positive breast cancer cells,and the underlying mechanism may be related to the blockage of proliferation-associated signaling pathways and the induction of apoptosis and cell cycle arrest.
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ObjectiveThrough the visual analysis of animal experimental literature on Traditional Chinese Medicine (TCM) nursing technology, the relevant research hot spots were condensed and the research trends of relevant animal models were discussed, to provide reference for subsequent research. MethodsThe literature related to animal experiments on Chinese medicine nursing technology was retrieved from Wan Fang database, Chinese Science and Technology Journal Database (VIP), China National Knowledge Infrastructure Engineering Database (CNKI), China Biomedical Literature Database (CBM), PubMed and Web of Science until June 30, 2022. Visual analysis was performed using VOSviewer 1.6.17 software. ResultsA total of 1 864 articles in Chinese and 126 articles in English were included, with the number of annual publications increasing year by year. The relevant literature involved 18 TCM nursing techniques, with the largest number (426) involving massage having the highest number of articles. It involved 4 496 authors, out of whom 358 were core authors accounting for 7.9% of all authors, and had a total count of 3 706 keywords forming 7 clusters. The research hotspots mainly included massage treatment of inflammatory diseases and analgesic effect, acupoint injection treatment of allergic rhinitis and myocardial ischemia, acupoint application treatment of asthma-related respiratory diseases, and moxibustion treatment of inflammatory diseases. The study of the mechanism of abdominal massage on insulin resistance is the latest research topic among them. ConclusionIn recent years, the animal model of abdominal massage has gained increasing popularity in animal experiments of TCM nursing techniques. However, the establishment and application of animal models related to Chinese medicine soaking and Chinese medicine hot ironing have not yet received attention. This area can be explored in the future to further improve the animal experimental research on Chinese medicine nursing technology.
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Improving the reproducibility of biomedical research results is a major challenge.Transparent and accurate reporting of the research process enables readers to evaluate the reliability of the research results and further explore the experiment by repeating it or building upon its findings. The ARRIVE 2.0 guidelines, released in 2019 by the UK National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs), provide a checklist applicable to any in vivo animal research report. These guidelines aim to improve the standardization of experimental design, implementation, and reporting, as well as the reliability, repeatability, and clinical translatability of animal experimental results. The use of ARRIVE 2.0 guidelines not only enriches the details of animal experimental research reports, ensuring that information on animal experimental results is fully evaluated and utilized, but also enables readers to understand the content expressed by the author accurately and clearly, promoting the transparency and integrity of the fundamental research review process. At present, the ARRIVE 2.0 guidelines have been widely adopted by international biomedical journals. This article is a Chinese translation based on the best practices of international journals following the ARRIVE 2.0 guidelines in international journals, specifically for the complete interpretation of the ARRIVE 2.0 guidelines published in the PLoS Biology journal in 2020 (original text can be found at https://arriveguidelines.org). The fourth part of the article includes the items 1-5 of ARRIVE 2.0 Recommended 11 section, which covers "Abstract" "Background" "Objectives" "Ethical statement" and "Housing and husbandry". Its aim is to promote the full understanding and use of the ARRIVE 2.0 guidelines by domestic researchers, enhance the standardization of experimental animal research and reporting, and promote the high-quality development of experimental animal technology and comparative medicine research in China.
ABSTRACT
Improving the reproducibility of biomedical research results is a major challenge.Researchers reporting their research process transparently and accurately can help readers evaluate the reliability of the research results and further explore the experiment by repeating it or building upon its findings. The ARRIVE 2.0 guidelines, released in 2019 by the UK National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs), provide a checklist applicable to any in vivo animal research report. These guidelines aim to improve the standardization of experimental design, implementation, and reporting, as well as the reliability, repeatability, and clinical translatability of animal experimental results. The use of ARRIVE 2.0 guidelines not only enriches the details of animal experimental research reports, ensuring that information on animal experimental results is fully evaluated and utilized, but also enables readers to understand the content expressed by the author accurately and clearly, promoting the transparency and integrity of the fundamental research review process. At present, the ARRIVE 2.0 guidelines have been widely adopted by international biomedical journals. This article is a Chinese translation based on the best practices of international journals following the ARRIVE 2.0 guidelines in international journals, specifically for the complete interpretation of the ARRIVE 2.0 guidelines published in the PLoS Biology journal in 2020 (original text can be found at https://arriveguidelines.org). The third part of the article includes the items 8-10 of ARRIVE 2.0 Essential 10, which covers "experimental animals" "experimental procedures" and "results". Its aim is to promote the full understanding and use of the ARRIVE 2.0 guidelines by domestic researchers, enhance the standardization of experimental animal research and reporting, and promote the high-quality development of experimental animal technology and comparative medicine research in China.
ABSTRACT
Improving the reproducibility of biomedical research results remains a major challenge. Transparent and accurate reporting of progress can help readers evaluate the reliability of research results and further explore an experiment by repeating or building upon its findings. The ARRIVE 2.0 guidelines, released in 2019 by the UK National Centre for the Replacement, Refinement, and Reduction of Animals in Research (NC3Rs), provide a checklist applicable to any in vivo animal research report. These guidelines aim to improve the standardization of experimental design, implementation, and reporting, as well as the reliability, repeatability, and clinical translatability of animal experimental results. The use of the ARRIVE 2.0 guidelines not only enriches the details of animal experimental research reports, ensuring that information on animal experimental results is fully evaluated and utilized, but also enables readers to understand the content expressed by the author accurately and clearly, promoting the transparency and integrity of the fundamental research review process. At present, the ARRIVE 2.0 guidelines have been widely adopted by international biomedical journals. This article is the second part of the Chinese translation of the complete interpretation of the ARRIVE 2.0 guidelines published in PLoS Biology in 2020 (original text can be found at https://arriveguidelines.org) and based on the best practices for following the ARRIVE 2.0 guidelines in international journals. This part includes Items 4-7 of "ARRIVE Essential 10" in the ARRIVE 2.0 guidelines: "Randomization", "Blinding", "Outcome Measurement", and "Statistical Methods". Our Chinese translated version aims to promote the full understanding and use of the ARRIVE 2.0 guidelines by domestic researchers, enhancing the standardization of experimental animal research and reporting, and promoting the high-quality development of experimental animal technology and comparative medicine research in China.
ABSTRACT
Influenza is a highly contagious disease that mainly affects the respiratory system and often leads to lung complications. Also it can cause a variety of very rare and serious neurological complications, including Guillain-Barre syndrome, transverse myelitis, meningoencephalitis and others. In recent years, neurological complications caused by influenza A virus have been reported in many countries and regions, and gradually attracted international attention. However, the pathogenesis of this complication remains unclear, and there are few related cases and animal experimental studies,and no specific treatment. Therefore, the authors summarized the study of neurological complications caused by influenza A virus in human and laboratory animals, in order to have a comprehensive understanding of the neurological diseases caused by influenza A virus.
ABSTRACT
Improving the reproducibility of biomedical research results is a major challenge. Researchers reporting their research process transparently and accurately can help readers evaluate the reliability of the research results and further explore the experiment by repeating it or building upon its findings. The ARRIVE 2.0 guidelines, released in 2019 by the UK National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs), provide a checklist applicable to any in vivo animal research report. These guidelines aim to improve the standardization of experimental design, implementation, and reporting, as well as the reliability, repeatability, and clinical translatability of animal experimental results. The use of ARRIVE 2.0 guidelines not only enriches the details of animal experimental research reports, ensuring that information on animal experimental results is fully evaluated and utilized, but also enables readers to understand the content expressed by the author accurately and clearly, promoting the transparency and integrity of the fundamental research review process. At present, the ARRIVE 2.0 guidelines have been widely adopted by international biomedical journals. this article is a Chinese translation based on the best practices of international journals following the ARRIVE 2.0 guidelines in international journals, specifically for the complete interpretation of the ARRIVE 2.0 guidelines published in the PLoS Biology journal in 2020 (original text can be found at https://arriveguidelines.org). The first part of the article includes the preface and the "Key 10" section, which covers "study design" "sample size" and "inclusion and exclusion criteria". Its aim is to promote the full understanding and use of the ARRIVE 2.0 guidelines by domestic researchers, enhance the standardization of experimental animal research and reporting, and promote the high-quality development of experimental animal technology and comparative medicine research in China.