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Objective:To discuss the effect of downregulating of high mobility group box protein 2(HMGB2)expression on the biological behavior of the liver cancer cells and the epithelial-mesenchymal transition(EMT)process,and to clarify its mechanism.Methods:The human liver cancer LM3 cells at logarithmic growth phase were divided into negative control group and HMGB2 RNA interference group(HMGB2 siRNA group);the cells in two groups were transfected with RNA oligonucleotides(RNA oligos)with irrelevant sequences and RNA oligos designed to knock down HMGB2,and the Lipofectamine 2000 was regarded as the vector.The expression levels of HMGB2 mRNA and protein in the cells in two groups were detected by real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting methods;cell scratch assay and Transwell chamber assay were used to detect the migration and invasion abilities of the cells in two groups;the expression levels of E-cadherin,N-cadherin,and Vimentin proteins and protein kinase B(AKT)/mammalian target of rapamycin(mTOR)pathway related proteins in the cells in two groups were detected by Western blotting method.Results:Compared with negative control group,the expression levels of HMGB2 mRNA and protein in the cells in HMGB2 siRNA group were significantly decreased(P<0.05),the cell scratch healing rate was significantly decreased(P<0.01),the number of invasion cells was significantly decreased(P<0.01),and the expression level of E-cadherin protein in the cells was significantly increased(P<0.01),while the expression levels of N-cadherin,Vimentin,mTOR,AKT,and phosphorylated AKT(p-AKT)proteins in the cells were significantly decreased(P<0.05 or P<0.01).Conclusion:Downregulating the expression of HMGB2 can reduce the migration and invasion abilities of the liver cancer LM3 cells and inhibit the EMT,and its mechanism may be related to regulating the expression of the AKT/mTOR pathway related proteins.
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The analysis of ammonia nitrogen in real water samples is challenging due to matrix interferences and difficulties for rapid on-site analysis.On the basis of the standard method,i.e.water quality-determination of ammonia nitrogen-salicylic acid spectrophotometry(HJ 536-2009),a simple device for online detecting ammonia nitrogen was developed using a sequential injection analysis(SIA)system in this work.The ammonia nitrogen transformation system,color reaction system,and detection system were built in compatible with the SIA system,respectively.In particular,the detection system was assembled by employing light-emitting diode as the light source,photodiode as the detector,and polyvinylchloride tube as the cuvette,thus significantly reducing the volume,energy consumption and fabricating cost of the detection system.As a result,the accurate analysis of ammonia nitrogen in complex water samples was achieved.A quantitative detection of ammonia nitrogen in water sample was obtained in 12 min,along with linear range extending to 1000 μmol/L,precisions(Relative standard deviation,RSD)of 4.3%(C=10 μmol/L,n=7)and 4.2%(C=500 μmol/L,n=7),and limit of detection(LOD)of 0.65 μmol/L(S/N=3,n=7).The results of interfering experiments showed that the detection of ammonia nitrogen by the developed device was not interfered by the common coexisting ions and components,therefore the environmental water could be directly analyzed,such as reservoir water,domestic sewage,sea water and leachate of waste landfill.The analytical results were consistent with those obtained by the environmental protection standard method(Water quality determination of ammonia nitrogen-salicylic acid spectrophotometry,HJ 536-2009).In addition,the spiking recoveries were in the range of 92.3%-98.1%,further confirming the accuracy and practicality of the developed device.
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Objective:To compare the difference between the operation of ureteral calculi patients with COVID-19 infection during COVID-19 infection and after recovery to find out the best timing for surgery.Methods:The clinical data of 327 ureteral calculi patients with COVID-19 infection were retrospectively analyzed, including 141 cases who underwent ureteroscopic surgery. According to the time of ureteroscopic surgery, the patients were divided into two groups: operation before recovery of COVID-19 infection group ( n=81) and operation after recovery of COVID-19 infection group ( n=60). The success rate of primary surgery, stone removal incidence, complication inciednce, mortality, operation time, postoperative hemoglobin decline, hospital stay, and total hospitalization cost and other indicators of the two groups were compared between the two groups. Measurement data were expressed as mean±standard deviation ( ± s), independent t-test was used for inter-group comparison. Chi-test or continuous correction Chi-test was used for inter-group comparison for count data. Results:The success rate of primary surgery (38.3%, 31/81), stone removal rate (86.4%, 70/81) in the operation before recovery of COVID-19 infection group were significantly lower than those in the operation after recovery of COVID-19 infection group [80.0% (48/60), 98.3% (59/60)], and the differences were statistically significant ( P< 0.05). The incidence of complication, postoperative hemoglobin decline [(2.1±0.6) g/L vs (0.6±0.3) g/L], hospital stay [(14.7±3.6) d vs (4.1±1.1) d], total hospitalization cost [(34 733.3±4 412.4) yuan vs (21 919.7±3 251.3) yuan] of the operation before recovery of COVID-19 infection group were significantly higher than those in the operation after recovery of COVID-19 infection group, and the differences were statistically significant ( P< 0.001). The operation time of the operation before recovery of COVID-19 infection group was shorter [(16.9±5.4) min vs (37.7±8.9) min], and the difference was statistically significant ( P< 0.001). There was no significant difference in mortality between the two groups (3.7% vs 0, P> 0.05). Conclusion:The risk of surgical treatment for ureteral calculi patients with COVID-19 infection is high, The treatment of them should mainly be conservative treatment, and operation should be performed after COVID-19 infection is cured.
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Objective To analyze the effect of LncRNA MEG3 on the blood-brain barrier in neonatal mice with hypoxic-ischemic brain injury(HIBD)by regulating miR-17-5p.Methods A total of 90 C57BL mice were randomly grouped into control group,model group,si-NC group,si-LncRNA MEG3 group,si-LncRNA MEG3+anti-miR-NC group and si-LncRNA MEG3+anti-miR-17-5p group,with 15 mice in each group.Except for the control group,the rest of the mice were constructed with HIBD model,and the Longa score was used to evaluate the nerve damage of mice.The ultrastructure of vascular endothelial cells around brain tissue was observed by electron microscope.The permeability of blood-brain barrier in mice was measured by EB method.The levels of LncRNA MEG3 and miR-17-5p in the brain tissue of mice in each group were determined by qRT-PCR.The levels of ZO-1 and Occludin protein in the brain tissue of mice were determined by Western blotting.Results Compared with control group,the vascular endothelium of mice in model group was unevenly thin and thick,and showed edema in many places.Compared with model group,the vascular endothelial cells in the si-LncRNA MEG3 group were gradually tightly connected,the thickness was more uniform,and the edema was reduced.Compared with the si-LncRNA MEG3 group,vascular endothelial cell damage was intensified in the si-LncRNA MEG3+anti-miR-17-5p group.Compared with those in the control group,the neurological deficit score,brain water content,EB content,and LncRNA MEG 3 level in the model group were significantly increased,miR-1 7-5 p level,ZO-1 and Occludin expression were significantly decreased(all P＜0.05).Compared with those in model group,there were no significant differences in neural function deficit score,brain water content,EB content,lncRNA MEG3 level,miR-17-5p level,ZO-1 and Occludin expression in si-NC group(all P＞0.05);the neurological deficit score,brain water content,EB content and LncRNA MEG3 level in si-LncRNA MEG3 group were significantly decreased,miR-17-5p level,ZO-1,and Occludin expression were significantly increased(all P＜0.05).Compared with those in si-LncRNA MEG3 group,there were no significant differences in neural function deficit score,brain water content,EB content,LncRNA MEG3 level,miR-17-5p level,ZO-1 and Occludin expression in si-LncRNA MEG3+anti-miR-NC group(all P＞0.05);the neurological deficit score,brain water content and EB content in si-LncRNA MEG3+anti-miR-17-5p group were significantly increased,miR-17-5p level,ZO-1 and Occludin expression were significantly decreased(all P＜0.05).Conclusion LncRNA MEG3 silencing may reduce the permeability of the blood-brain barrier in neonatal mice with HIBD by up-regulating miR-17-5p,maintaining the protective effect of the blood-brain barrier on the brain,and protecting brain tissue.
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Objective To construct a measurement tool for atrial fibrillation(AF)patients'experience of catheter ablation,in order to provide quantifiable basis for improving the patients'perioperative experience.Methods From Jun 2022 to Apr 2023,literature analysis,qualitative research,Delphi expert consultation,and analytic hierarchy process were used to determine the content and weight of various indicators of the measurement tool.Results The enthusiasm of experts in 3 rounds was 100%.The authority coefficient of experts was 0.946,0.961 and 0.976.The Kendal harmony coefficients of the 2 and 3 rounds of expert consultation was 0.130 and 0.370(P＜0.001).The final measurement tool included 46 items and 5 dimensions,including operational and technical quality experience,comfort management experience,information and communication experience,emotional support experience,service process and response experience.Conclusion The preliminary construction of measurement tool for AF patients'experience of catheter ablation,which were based on the features of specialty,could not only evaluate the patients'experience accurately,but also provide a basis for targeted improvement of medical and nursing service quality.
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BACKGROUND:Functional training has been popular in recent years,but it is mainly applied in sports training field.There are still insufficient studies and applications in medical and health fields. OBJECTIVE:To provide a theoretical basis for relevant research in sports,medical and health fields,through a more comprehensive and in-depth exploration and analysis of the research hot spots,ideological trends,frontiers and development trends of international functional training in the field of medical and health care. METHODS:The 2 206 high-quality articles addressing health-related functional training during 2012-2022 were exported from the Web of Science Core Set Database as the object of analysis.Combined with research methods such as literature analysis,Citespace V analysis software was used for visual analysis of keywords,subject categories and highly cited literatures. RESULTS AND CONCLUSION:The number of articles published on functional training in the field of health is on the rise.There are more articles from the United States,with a larger impact.China also has a high volume of publications,but the impact and depth of research is lacking.Improving physical and mental health and cognitive ability of middle-aged and older people is the main focus,followed by preventing sports injuries and promoting recovery in athletes.In the future,more research will be conducted on teenagers,the disabled and other groups,and there will be a continued increase in injury prevention and recovery promotion for athletes.Chinese scholars have less research on the effects of functional training on the physical health of the general public,and more attention should be paid to improving the physical and mental health of the general public.
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Objective:To study the clinical characteristics of congenital esophageal atresia (CEA) and risk factors of mortality associated with esophageal repair (ER) surgery.Methods:From January 2010 to December 2022, patients diagnosed of CEA using chest and abdomen X-ray and esophagography in our hospital were retrospectively reviewed. The patients were assigned into ER group and non-ER group according to the treatments. The ER group was subgrouped into survival group and death group according to the prognosis. Clinical data and outcomes were collected and compared between the groups.Results:A total of 553 cases were enrolled. According to Gross classification, 29 patients (5.2%) were type A, 2 patients (0.4%) were type B, 504 patients (91.1%) were type C, 6 patients (1.1%) were type D and 11 patients (2.0%) were type E. One patient had simple transluminal septal atresia of the esophagus. 406 patients were in ER group and 147 in non-ER group. Compared with ER group, non-ER group had significantly higher incidences of preterm birth, low birth weight and overall malformations (all P<0.05). In ER group, 152 patients (37.4%) received open thoracic surgery (OTS), 243 (59.9%) had video-assisted thoracoscopic surgery (VATS) and 11 (2.7%) were VATS converted to OTS. Postoperative anastomotic leakage (PAL) occurred in 92 patients (22.7%) and 15 patients (3.7%) died after surgery. The median length of hospital stay was 23 (17, 36) d. Compared with the survival group, the death group had higher incidences of preterm birth, low birth weight, VATS converted to OTS, mechanical ventilation after ER, and shorter length of hospital stay (all P<0.05). After adjusted for birth weight, VATS converted to OTS ( OR=9.585, 95% CI 1.899-48.374) and mechanical ventilation after ER ( OR=7.821, 95% CI 1.002-61.057) were risk factors of mortality in ER patients. Conclusions:Non-ER patients have higher incidences of preterm birth, low birth weight and overall malformations than ER patients. VATS is the method of choice for CEA. Preterm birth, low birth weight, VATS converted to OTS and mechanical ventilation after ER are risk factors of mortality in ER patients.
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Objective To investigate the effects of circDNMT1 on the proliferation,apoptosis and epithelial-mesenchymal transition(EMT)of triple-negative breast cancer(TNBC)cells by regulating miR-377-3p/PUM1 axis.Methods The TNBC tissues and adjacent normal tissues of 24 patients with TNBC treated in Danzhou People's Hospital from 2018 to 2021 were collected.qRT-PCR and Western blot were used to detect the expression of circDNMT1,miR-377-3p,and PUM1 protein in tissue and mouse normal breast epithelial cell line HC11 and TNBC cell lines 4T1,Eph41424,and JC.4T1 cells were divided into the 4T1 group(untransfected),the si-NC group(transfected with si-NC),the si-DNMT1 group(transfected with si-DNMT1),the si-DNMT1+anti-NC group(simultaneously transfected with si-DNMT1 and anti-NC),and the si-DNMT1+anti-miR-377-3p group(simultaneously transfected with si-DNMT1 and anti-miR-377-3p).qRT-PCR was used to detect the expression of circDNMT1 and miR-377-3p of 4T1 cells in each group;CCK-8 was used to detect the proliferation of 4T1 cells in each group;flow cytometry was used to detect the apoptosis of 4T1 cells in each group;Western blot was used to detect the expression of PUM1,EMT-related proteins and apoptosis-related proteins of 4T1 cells in each group;TargetScan website was used to predict the binding sites of miR-377-3p with circDNMT1 and PUM1;dual luciferase report was used to verify the targeting relationships of miR-377-3p with circDNMT1 and PUM1.After inoculation with 4T1 cells,BALB/c mice were randomly divided into the blank control group(injected with equal amount of normal saline),the negative control group(injected with si-NC via tail vein),the DNMT1 silencing group(injected with si-DNMT1 via tail vein),the combined control group(injected with si-DNMT1 and anti-NC via tail vein),and the combined silencing group(injected with si-DNMT1 and anti-miR-377-3p via tail vein),the tumor massess of mice were recorded and the morphological changes of tumors were observed by hematoxylin-eosin staining.Results circDNMT1 and PUM1 were up-regulated in TNBC tissues and cells,and miR-377-3p was down-regulated.The expression difference was most obvious in 4T1 cells,so 4T1 cells were selected for subsequent experiments.Compared with the 4T1 group and the si-NC group,the expression of miR-377-3p,the apoptosis rate of 4T1 cells,the expression levels of Bax,cleaved caspase-3 and E-cadherin protein of 4T1 cells in the si-DNMT1 group were significantly increased(P<0.05),the circ-DNMT1 level,the expression level of PUM1 protein,OD values at 24 hours and 48 hours,the expression level of Bcl-2,N-cadherin,Vimentin protein were significantly decreased(P<0.05).Compared with the si-DNMT1 group and the si-DNMT1+anti-NC group,the expression of miR-377-3p,the apoptosis rate of 4T1 cells,the expression levels of Bax,cleaved caspase-3 and E-cadherin proteins of 4T1 cells in the si-DNMT1+anti-miR-377-3p group were significantly decreased(P<0.05),the expression level of PUM1 protein,OD values at 24 hours and 48 hours,the expression levels of Bcl-2,N-cadherin,Vimentin proteins were significantly increased(P<0.05).Compared with the miR-NC+WT-circDNMT1 group,the cell luciferase activity in the miR-377-3p mimic+WT-circDNMT1 group was significantly decreased(P<0.05);compared with the miR-NC+WT-PUM1 group,the cell luciferase activity in the miR-377-3p mimic+WT-PUM1 group was significantly decreased(P<0.05).The tumor cells in the blank control group and the negative control group were densely arranged with clear boundary;the tumor cells in the DNMT1 silencing group and the combined control group were loosely arranged,the nuclei were pyknotic,and the cell fragments were increased;the tumor cells in the combined silencing group were densely arranged and the boundaries tended to be clear.Compared with the blank control group and the negative control group,the tumor mass of mice in the DNMT1 silencing group was significantly decreased(P<0.05).Compared with the DNMT1 silencing group and the combined control group,the tumor mass of mice in the combined silencing group was significantly increased(P<0.05).Conclusion Silencing circDNMT1 may inhibit the expression of PUM1 by up-regulating miR-377-3p,thereby inhibiting the proliferation and EMT of TNBC cells,and promoting cell apoptosis.
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Objective:To analyze the key points and shortcomings of Traditional Chinese Medicine(TCM)science and technology innovation policy in China,and to provide reference for the subsequent policy optimization.Methods:Searching for TCM science and technology innovation policy texts released at the national level since 2007,and use the two-dimensional analysis framework for quantitative analysis.Results:Among 27 policies,In the X dimension,supply-based,environmental and demand-based policy tools respectively accounted for 48.98%,39.29%and 11.73%.In the Y dimension,the proportion of scientific and technological innovation and achievement transformation was the highest(29.03%);Promoting the development of integrated Chinese and Western medicine was the least used(1.08%).Conclusion:There is a structural imbalance in the application of policy tools,the distribution difference of policy objectives is significant,and the internal policy tools'usage is imbalanced regarding policy objectives.Suggestions:Optimize the internal structure of policy tools.Meanwhile,enhance the structure of policy objectives,and facilitate the dynamic integration and application of policy tools to achieve the policy objectives of scientific and technological innovation in TCM.
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OBJECTIVE@#To observe the alteration of thoracic and lumbar physiological curvature in adolescent idiopathic scoliosis(AIS) and the difference of physiological curvature between different types of scoliosis.@*METHODS@#A retrospective analysis was conducted on 305 adolescent patients taken full spine X-ray in our hospital from January 2017 to December 2021. The patients were divided into normal group and scoliosis group. The normal group was composed of 179 patients, 79 males and 100 females, aged 10 to 18 years old with an average of (12.84±2.10) years old, with cobb agle less than 10 degrees. The scoliosis group was composed of 126 patients, 33 males and 93 females, aged 10 to 18 years old with an average of (13.92±2.20) years old. The gender, age, Risser sign, thoracic kyphosis(TK) and lumbar lordosis(LL) in 2 groups were compared, and the TK and LL were also compared between different genders, different degrees of scoliosis and different segments of scoliosis.@*RESULTS@#The female ratio(P=0.001) and age (P<0.001) in scoliosis group were higher than them in normal group; the ratio of low-grade ossification was higher in normal group than in scoliosis group(P=0.038). TK was significantly smaller in scoliosis group than in normal group(P<0.001), but there was no significant difference in LL between the 2 groups(P=0.147). There were no significant difference in TK and LL between male and female. The TK was significantly bigger in mild AIS patients than in moderate AIS patients(P<0.05), but there was no significant difference in LL between mild and moderate patients(P>0.05). The TK and LL in different segments scoliosis were not found significant difference.@*CONCLUSION@#The physiological curvature of thoracic and lumbar spine is independent of gender. The thoracic physiological curvature becomes smaller in AIS patients, but lumbar curvature remains unchanged. The thoracic physiological curvature in mild AIS patients is greater than that in moderate AIS patients, but the lumbar curvature is almost unchanged between mild and moderate scoliosis and is similar with that in normal adolescent. The alteration of thoracic and lumbar physiological curvature in AIS patients may be related to relative anterior spinal overgrowth, and the specific detailed mechanism needs to be further studied.
الموضوعات
Female , Humans , Male , Adolescent , Child , Scoliosis/diagnostic imaging , Retrospective Studies , Thoracic Vertebrae/diagnostic imaging , Kyphosis , Lordosis , Lumbar Vertebrae/diagnostic imaging , Spinal Fusion/methodsالملخص
ObjectiveTo evaluate the clinical efficacy and safety of Ganjie Bingmei Tablet （甘桔冰梅片， GBT） in treating acute pharyngitis with wind-heat invading the lung syndrome. MethodsUsing multi-center， rando-mized， double-blind， double-dummy， positive and parallel controlled clinical trial design， 144 acute pharyngitis patients with wind-heat invading the lung syndrome from October 8th， 2022 to March 31st， 2023 were collected and randomly assigned to the treatment group and the control group， with 72 cases in each group. The treatment group was treated by GBT （0.4 g each time， 4 times a day） combined with Ganjie Qingyan Granules （甘桔清咽颗粒， GQG） placebo （10 g each time， 3 times a day）， while the control group was treated by GQG （10 g each time， 3 times a day） combined with GBT placebo （0.4 g each time， 4 times a day）， both for 5 days. The clinical efficacy of the two groups was observed， and the disappearance rate of pharyngalgia and the efficacy regarding TCM syndromes were compared between groups after treatment. The visual analog scale （VAS） score before and after treatment was assessed， and the safety was evaluated. ResultsThe disappearance rate of pharyngalgia in the treatment group was 98.61%（71/72）， significantly higher than 80.56%（58/72） in the control group （P＜0.01）. The VAS scores in both groups significantly decreased after treatment （P＜0.05）， and lower score was found in the treatment group than the control group （P＜0.05）. The total effective rate regarding TCM syndromes was 100% in both groups， but the curative rate was significantly higher in the treatment group（73.61%， 53/72） than the control group （62.50%， 45/72， P＜0.05）. There were no obvious adverse reactions in the both groups. ConclusionGBT is effective and safe in treating acute pharyngitis with wind-heat affecting the lung syndrome.
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ObjectiveTo investigate the impact of icariin （ICA） on autophagy in glucocorticoid-induced bone microvascular endothelial cells （BMECs） mediated by the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin （PI3K/Akt/mTOR） signaling pathway. MethodBMECs were isolated and cultured from femoral heads obtained during total hip arthroplasty and identified using immunofluorescence staining. The experimental cells were divided into four groups： A control group， a glucocorticoid group （100 mg·L-1 hydrocortisone）， an ICA group （100 mg·L-1 hydrocortisone+6.7×10-3 mg·L-1 ICA）， and a Rapamycin group （100 mg·L-1 hydrocortisone+6.7×10-3 mg·L-1 ICA+1 mg·L-1 rapamycin）. Autophagy in BMECs was induced using 100 mg·L-1 hydrocortisone. LC3 fluorescence staining was used to observe the peak of autophagy at different time points. Western blot analysis was employed to analyze the expression of autophagy-related proteins and PI3K/Akt/mTOR pathway proteins in each group. Electron microscopy was used to observe autophagosomes and autolysosomes in the cells. ResultHydrocortisone at 100 mg·L-1 induced autophagy in BMECs， reaching a peak at around 5 hours， which then declined with further intervention. Compared to the control group， the glucocorticoid group showed cell membrane damage， disordered organelle arrangement， and a large number of autophagosomes and autolysosomes. Compared to the glucocorticoid group， the ICA group had more intact cell membranes， sparser organelle arrangement， and fewer autophagosomes and autolysosomes. Compared to the ICA group， the Rapamycin group showed cell membrane damage， disordered organelle arrangement， and more autophagosomes and autolysosomes. Compared to the control group， the glucocorticoid group had significantly increased expression of light chain 3B （LC3B）， Atg4B， and p62 （P<0.01）. Compared to the glucocorticoid group， the ICA group showed significantly decreased expression of LC3B， Atg4B， p62， and Beclin-1 （P<0.01）. Compared to the ICA group， the Rapamycin group had significantly increased expression of Atg4B and p62 （P<0.01）. Compared to the control group， the glucocorticoid group had significantly decreased expression of p-PI3K/PI3K， p-Akt/Akt， and p-mTOR/mTOR （P<0.01）. Compared to the glucocorticoid group， the ICA group showed significantly increased expression of p-PI3K/PI3K， p-Akt/Akt， and p-mTOR/mTOR （P<0.01）. Compared to the ICA group， the Rapamycin group had significantly decreased expression of p-PI3K/PI3K， p-Akt/Akt， and p-mTOR/mTOR （P<0.01）. Ubiquitination levels were significantly decreased in the glucocorticoid group compared to the control group （P<0.01）. Compared to the glucocorticoid group， ubiquitination levels were significantly increased in the ICA group （P<0.01）， and significantly decreased in the Rapamycin group compared to the ICA group （P<0.01）. ConclusionThe glucocorticoid-induced autophagy in BMECs is time-dependent. ICA inhibits glucocorticoid-induced autophagy in BMECs， and this effect may be related to the regulation of the PI3K/Akt/mTOR pathway.
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The microbiological quality of laboratory animals is crucial for the validity and reproducibility of scientific research data, as well as human health and animal welfare. Currently, individual ventilation cages (IVC) have become the mainstream feeding system for rodent laboratory animals. The most commonly used pathogen monitoring method for this feeding system is soiled bedding sentinels (SBS). This method monitors the microbial carrying status of mouse colony through indirect contact and delayed feedback. It can effectively monitor pathogens transmitted via the fecal-oral route, such as mouse hepatitis virus and reovirus. However, this method has difficulty detecting pathogens mainly transmitted through aerosols or direct contact, such as Sendai virus and Pasteurella pneumotropica. The exhaust air dust (EAD)-PCR monitoring method involves swab sampling in the IVC exhaust ducts to monitor the corresponding racks of the ducts; swab sampling before the prefiltration of the host to monitor the entire IVC rack; and EAD collection device sampling to monitor all racks connected to the same host. Different IVC manufacturers have developed corresponding EAD collection devices for their respective IVC systems, making operations convenient and standardization easy. Compared with the SBS method, the EAD-PCR method significantly improves detection rate and timeliness, with the fastest detection possible after one week of exposure. It can serve as a supplement or replacement for the SBS method. Currently, increasing evidence supports that EAD-PCR testing is a more reliable, sensitive, and cost-effective monitoring method, and is more beneficial to animal welfare. This article reviews the application progress of these two methods for monitoring pathogens, analyzes the existing limitations of the EAD-PCR method, and proposes solutions based on its implementation in our laboratory and examination units. The EAD-PCR method helps reduce the number of live sentinel animals used in pathogen monitoring, in order to better maintain the "3Rs" principle of laboratory animal welfare.
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ObjectiveTo explore whether miR-375 regulates the malignant characteristics of osteosarcoma (OS) by influencing the expression of MMP13. MethodsPlasmid DNAs and miRNAs were transfected into OS cells and HEK293 cells using Lipofectamine 3000 reagent. Real-time quantitative polymerase chain reaction was performed to measure the expression of miR-375 and MMP13 in OS patients and OS cells. Western blot was performed to analyze the MMP13 protein in the patients with OS and OS cells. The targeting relationship between miR-375 and MMP13 was analyzed by luciferase assay. Migration and invasion were analysed by heal wound and transwell assays, respectively. ResultsmiR-375 expression in OS tissues was lower than that in normal tissues. The expression of MMP13 was upregulated in OS tissues. MMP13 expression was negatively correlated withmiR-375 expression in patients with OS. Migration and invasion were significantly inhibited in OS cells with the miR-375 mimic compared with OS cells with the miRNA control. MMP13 partially reversed the inhibition of migration and invasion induced by miR-375 in the OS cells. ConclusionmiR-375 attenuates migration and invasion by downregulating the expression of MMP13 in OS cells.
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Ubiquitination, a diverse post-translational modification, is carried out by enzymes including E1-activating enzymes, E2-conjugating enzymes, E3 ligases, and deubiquitinating enzymes (DUBs). Ubiquitin itself possesses 7 lysine residues and N-terminal methionine, allowing for the formation of polyubiquitin chains with different lengths and linkages. These chains exhibit various topologies that can be recognized by proteins containing ubiquitin-binding domain, thereby transmitting distinct cellular signals. To unravel the physiological mechanisms associated with ubiquitin, numerous ubiquitin probes have been developed. This review provides an overview of recent advancements in the field of ubiquitin probes, focusing on activity-based and affinity-based probes. Activity-based probes are designed to covalently bind to DUBs, E1s, or E3s, enabling the identification and characterization of these enzymes. Affinity-based probes, on the other hand, selectively bind to ubiquitin-binding domains, facilitating the identification of proteins that interact with ubiquitin. Moreover, this review comprehensively discusses the synthetic methodologies employed for the acquisition of ubiquitin probes. These includes meticulous discussions on the synthesis of individual monomeric modules, the establishment of isopeptide linkages, as well as the incorporation of reactive functional groups. Additionally, the review explores the emerging area of cell-penetrating ubiquitin probes and highlights their latest applications in living cells. These probes incorporate cell-penetrating peptides to enable their internalization into cells, allowing for direct visualization and manipulation of ubiquitin-modified proteins within their native environment. Overall, this review offers insights into the design, synthesis, and applications of ubiquitin probes, highlighting their significance in elucidating ubiquitin-mediated cellular processes.
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OBJECTIVE To investigate the efficacy and safety of piperacillin-tazobactam in the treatment of complicated urinary tract infection （cUTI） in adults. METHODS Retrospective analysis was performed on the data of 352 cUTI adult patients in our hospital from January 1， 2021 to December 31， 2023. All patients received piperacillin-tazobactam. The detection of pathogens in patients， the clinical efficacy and microbial clearance rate after treatment， the occurrence of adverse drug reactions and treatment cost were observed in all patients. RESULTS Of the 352 patients， pathogen culture results of 54 patients were detected， mainly Escherichia coli producing extended-spectrum beta-lactamases. The clinical effective rate was 94.3%， the microbial clearance rate was 81.5%， and the incidence of adverse reactions was 1.4%. The percentage of male effective patients in urinary surgery department was significantly higher than invalid patients， while the proportion of transplant treatment and the proportion of patients with concomitant kidney transplantation were significantly lower than invalid patients （P＜0.05）. There was no significant difference in clinical effective rate between the two groups after those patients were divided into target treatment group and empirical treatment group according to the sensitivity of pathogen to piperacillin-tazobactam （P＝0.902 5）. CONCLUSIONS Piperacillin-tazobactam is effective and safe in the treatment of cUTI.
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Thirty-one phenolic constituents were isolated and purified from the 95% ethanol extract of Sanguisorbae Radix by using various chromatographic techniques, including macroporous resin, silica gel, ODS, Sephadex LH-20 and semi-preparative HPLC. Their structures were elucidated by physicochemical properties, spectroscopic data (MS and NMR) and electronic circular dichroism (ECD) spectra, and identified as 3-methoxyl-2S,3S-epoxyflavanone (1a), 3-methoxyl-2R,3R-epoxyflavanone (1b), longifoin B (2), longifoin C (3), eriodictyol (4), naringenin (5), liquiritigenin (6), 5,3ʹ-dihydroxy-7,4ʹ-dimethoxyflavanone (7), naringenin-7-O-β-D-glucopyranoside (8), dihydroquercetin (9), dihydrokaempferol (10), (-)-garbanzol (11), (2R,3R)-4-methoxyl-distylin (12), kaempferol (13), quercetin (14), α,4,2′,4′-tetrahydroxydihydrochalcone (15), phloretin (16), (+)-catechin (17), ethyl (+)-cyanidan-3-ol-8-carboxylate (18), phyllocoumarin (19), methyl 3-methoxy-4,5-dihydroxybenzoate (20), 4,5-dimethoxy-3-hydroxybenzoic acid methyl ester (21), 3,4′-di-O-methylellagic acid (22), 3,4,3′-O-trimethylellagic acid (23), 3,3ʹ,4ʹ-O-trimethylellagic acid-4-O-β-D-xyloside (24), (3R)-thunberginol C (25), resveratrol (26), 1-hydroxypinoresinol (27), (7S,8S)-3-methoxy-3′,7-epoxy-8,4′-oxyneoligna-4,9,9′-triol (28), emodin-8-O-β-D-glucoside (29), phloracetophenone (30) and 4-(4′-hydroxyphenyl)-butan-2-one (31). Among them, compound 1a and 1b is a pair of new flavonoid enantiomers, compounds 2 and 3 are a pair of new epimers, while compounds 4, 5, 6, 9, 10, 13, 16 and 26 were obtained from S. officinalis for the first time, compounds 7, 8, 27, 30 and 31 were isolated for the first time from the S. officinalis genus, and compounds 11, 12, 15, 18, 19, 25, 28 and 29 were isolated for the first time from the Rosaceae. The antioxidant activities of compounds 1-24 were evaluated by activating the Nrf2 transcriptional pathway, which were measured by the dual-luciferase reporter gene assay in 293T cells. Compounds 4, 6-10, 12, 14, 17, 19, 20 and 22-24 showed significant Nrf2 agonistic effect compared with the control group at 25 μmol·L-1, which provided reference for the research of their antioxidant activity.
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ObjectiveTo investigate the clinical efficacy of Qihuang Jianpi Zishen Granules in the treatment of systemic lupus erythematosus （SLE） and its effect on the signal transducer and activator of tranSCription 3/mammalian target of rapamycin （STAT3/mTOR） signaling pathway， and to decipher the possible mechanism. MethodSixty female SLE patients who met the criteria in the First Affiliated Hospital of Anhui University of Chinese Medicine from May 2022 to May 2023 were selected and randomized into a control group and an observation group （30 cases in each group）. The control group was treated with prednisone acetate + hydroxychloroquine sulfate orally， and the observation group was additionally treated with Qihuang Jianpi Zishen granules. The treatment lasted for 8 weeks. The SLE disease activity （SLEDAI）， TCM syndrome score， erythrocyte sedimentation rate （ESR）， hypersensitive C-reactive protein （hs-CRP）， immune indexes ［immunoglobulin G （IgG）， C3， C4， CD4+， and CD8+］， interleukin （IL）-17， IL-23， interferon （IFN）-γ， 24 h urinary protein （24 h PRO）， serum creatinine （SCr）， and expression of proteins ［STAT3， phosphorylated （p）-STAT3， mTOR protein and STAT3，mTOR mRNA］ in the STAT3/mTOR signaling pathway were determined before and after treatment. In addition， the adverse reactions were recorded. ResultAfter 8 weeks of treatment， the total response rate in the observation group was 93.33% （28/30）， which was higher than that （70.00%， 21/30） in the control group （χ2=4.007， P<0.05）. After treatment， both groups showed declined SLEDAI， TCM syndrome score， ESR， hs-CRP， IgG， CD8+， IL-17， IL-23， IFN-γ， 24 h PRO， SCr， and expression of proteins in the STAT3/mTOR pathway （P<0.01） and elevated levels of C3， C4， and CD4+ （P<0.01）. Moreover， the observation group had lower SLEDAI， TCM syndrome score， ESR， hs-CRP， IgG， CD8+， IL-17， IL-23， IFN-γ， 24 h PRO， SCr， and expression of proteins in the STAT3/mTOR pathway （P<0.05， P<0.01） and higher levels of C3， C4， and CD4+ （P<0.05， P<0.01） than the control group after treatment. Neither group showed serious adverse reactions during the treatment period. ConclusionQihuang Jianpi Zishen Granules can ameliorate the inflammatory response， reduce the disease activity， and mitigate the kidney injury in SLE by inhibiting the STAT3/mTOR signaling pathway to regulate the immune function.
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Objective:To analyze the effect of mucosal ligament preservation on the outcome of arthroscopic repair of meniscus injury.Methods:A total of 77 patients with knee meniscus tears caused by sports injuries who underwent arthroscopic meniscus repair or suture in Zhengzhou Orthopaedic Hospital from June 2022 to June 2023, were retrospectively analyzed. Including 45 males and 32 females, aged 38.57±13.54 years (range, 52-87 years), body mass index 24.72±4.01 kg/m 2 (range, 34.14-13.61 kg/m 2). All patients complained of knee pain, limited activity and walking weakness. The symptoms were not relieved after 1 month of conservative treatment, which seriously affected daily work and life. According to the intraoperative treatment of mucosal ligament, the patients were divided into mucosal ligament preservation group and mucosal ligament removal group. The visual analogue score (VAS), Lysholm score, and total blood loss were compared between the two groups. Results:All patients successfully completed the operation and were followed up for an average of 5.23±2.16 months (range, 3-9 months). The operation time was 47.59±16.81 min in mucosal ligament preservation group and 45.25±15.93 min in mucosal ligament removal group, and there was no significant difference between the two groups ( t=0.628, P=0.532). The total blood loss in the mucosal ligament preservation group was 246±193 ml, which was less than 343±211 ml in the mucosal ligament removal group, and the difference was statistically significant ( t=2.095, P=0.040). None of the patients received allogeneic blood transfusion. The hematocrit of the mucosal ligament preservation group was 42.48%±4.57% before operation and 39.42%±4.65% on the third day after operation, while that of the mucosal ligament removal group was 41.24%±4.16% and 38.95%±3.80%. The difference between the two groups was statistically significant ( t=0.016, P=0.004; t=0.004, P=0.016). There was no significant difference between the two groups before operation and on the third day after operation ( t=0.217, P=0.545; t=0.629, P=0.159). The preoperative VAS score of mucosal ligament preservation group was 7.25±1.10, which was higher than that of 3 months after operation (0.83±1.06), and the difference was statistically significant ( t=0.062, P<0.001). The preoperative VAS score of mucosal ligament removal group was 7.16±1.21, which was higher than that of 3 months after operation (1.05±1.13), and the difference was statistically significant ( t=0.017, P<0.001). There was no significant difference in VAS scores between the two groups before operation and at 3 months after operation ( t=0.144, P=0.740; t= 0.273, P=0.603). The preoperative Lysholm score of mucosal ligament preservation group was 31.76±7.54, which was significantly lower than that of 3 months after operation 87.30±4.12 ( t=-39.329, P<0.001); The Lysholm score of the mucosal ligament removal group was 34.13±7.32 before operation, which was lower than 89.05±4.45 at 3 months after operation, and the difference was statistically significant ( t=-40.172, P<0.001); There was no significant difference in Lysholm score between the two groups before operation and 3 months after operation ( t=1.395, P=0.167; t=1.766, P=0.081). Conclusion:The preservation of mucosal ligament in arthroscopic surgery for meniscus injury does not prolong the operation time. It can reduce the total intraoperative blood loss, and the postoperative knee function recovery is similar to that of mucosal ligament removal.
الملخص
Objective To investigate the effect of galangin on cardiac remodeling and cardiac func-tion after myocardial infarction(MI).Methods A total of 32 male C57/BL6 mice(8-10 weeks old)were subjected for MI modeling,and finally 24 mice were assigned into control group[sham operation+hydroxycellulose sodium(CMC-Na)],model group(MI+CMC-Na),and experimental group(MI+galangin),with 8 mice in each group.After MI modeling,the mice of the experimen-tal group were given 40 mg/kg galangin by gavage for 4 weeks,and those of the control group and the model group were given the same volume(0.4 ml)of CMC-Na solution.HE staining was used to observe the size of the infarct area.The mRNA levels of inflammatory factors in the heart were detected by qRT-PCR,and protein levels of related signaling pathway proteins were measured with Western blotting.Immunofluorescence(IF)assay was applied to detect the infiltration of in-flammatory cells in the infarct border zone.TUNEL staining was employed to detect cell apoptosis in the infarct border zone.Results At 4 weeks after modeling,larger infarct size,enhanced expression levels of IL-1β,IL-6,TNF-α,p-P65,p-IκBα and Bax,elevated apoptotic rate,decreased cardiac function indicators such as FS and LVEF,and reduced Bcl-2 expression level were observed in the model group than the control group(P＜0.05).The experimental group had sig-nificant smaller myocardial infarct size[(11.64±0.64)％vs(21.84±1.94)％],less CD3 positive T cells[(3.10±0.46)％vs(6.28±0.24)％],F4-80 positive macrophages[(1.98±0.50)％vs(5.35±0.62)％]and LY6G positive neutrophils[(6.33±0.67)％vs(11.33±1.77)％],decreased expression levels of IL-1β,IL-6,TNF-α,p-P65,p-IκBα and Bax,reduced apoptotic rate[(21.45± 1.62)％vs(35.68±0.88)％],and increased FS and LVEF values and Bcl-2 expression level when compared with the model group(P＜0.05).Conclusion Galangin improves myocardial remode-ling and cardiac dysfunction after MI by inhibiting inflammatory response and cell apoptosis.