1.Effect and Mechanism of Cyclocarya paliurus(Batal.)lljinskaja Polysaccharides on Type 2 Diabetic Rats by Regulating GLUT4 Translocation in Islet and Liver
Haiyun LIU ; Miaoting SHI ; Xinyi LUO ; Minyan SUN ; Chenxi XU ; Kunhan CHEN ; Xiaomin WANG ; Rengeng SHU
Herald of Medicine 2024;43(1):13-19
Objective To investigate the effect of Cyclocarya paliurus(Batal.)lljinskaja polysaccharides on insulin resistance in type 2 diabetic rats by regulating glucose transporter 4(GLUT4)translocation in islet and liver.Methods High-fat diet combined with low-dose streptozotocin(35 mg·kg-1)to induce type 2 diabetes model,all the rats were randomly divided into model control group,Cyclocarya paliurus polysaccharides groups(5,10 g·kg-1)and metformin group(0.25 g·kg-1),and treated for eight weeks(n=9 in each group).Fasting glucose and lipid were determined.Histopathology of rat islet and liver were observed by hematoxylin and eosin staining.Protein expressions of phosphorylated phosphoinositide-3-kinase(p-PI3K),phosphorylated serine-threonine kinase 1(p-Akt1),and GLUT4 in islet were measured by immunohistochemistry staining.GLUT4 translocation in the islet and liver was detected by immunofluorescence.Results Compared with the model control group,the Cyclocarya paliurus polysaccharides group and metformin group had declined fasting glucose levels and increased high-density lipoprotein(P<0.05).The structure of the islets and liver was relatively complete.The content of p-PI3K,p-Akt1 and GLUT4 in the islet increased(P<0.05).GLUT4 translocation in the liver and islet enhanced(P<0.05).Conclusions Cyclocarya paliurus polysaccharides alleviate glucose and lipid metabolism disorders.The mechanism may lay in it activating protein expressions of p-PI3K,p-Akt1,and GLUT4 in islet cells.GLUT4 translocation to the islet and liver cell membrane are increased to regulate peripheral islet resistance.
2.Establishment of internal quality control methodology for blood transfusion compatibility testing
Lu LI ; Xiaolin SUN ; Junjie WEI ; Ruiqi LIU ; Weixin WU ; Haiyun LIU ; Yinze ZHANG
Chinese Journal of Blood Transfusion 2024;37(4):399-404
【Objective】 To monitor the effectiveness and accuracy of the blood transfusion compatibility test system by self-made weakly positive internal quality control products. 【Methods】 Red blood cells from DAT(-) healthy subjects were selected, and B/RhD(-)E(-) red blood cells were selected as tube 1. A/RhD(+ )E(+ ) was selected as tube 2 to prepare blood group quality control products according to the principle of blood group antigen compatibility, and red blood cell preservation solution and corresponding ABO blood group reagent antibody were added to make the agglutination intensity of microcolumn gel method in reverse blood typing reach a low positive value (1+ ). Tube 3 and tube 4 were prepared with five different preservation media: plasma, serum, antibody diluent, mixture of equal plasma and antibody diluent, and mixture of equal serum and antibody diluent, respectively. IgM anti-E antibody was added to tube 3, and IgG anti-D antibody was added to tube 4, so that the agglutination intensity of microcolumn gel method reached a low positive value (1+ ). 【Results】 Comparison between the 5 different preservation media showed that the preservation medium of antibody diluent was the most stable for weakly positive antibody (F=11.35, P<0.05), Agglutination intensity 1+ is assigned 5 points by AABB Technical Manual, and its score was 5.25±1.75 points. 【Conclusion】 The use of self-made weakly positive quality control products can improve the effectiveness, accuracy and sensitivity of the monitoring system, thus achieving internal quality control and ensuring the safety of clinical blood use.
3.Effect of RhD antigen expression intensity on preparation of low positive quality control products for transfusion compatibility test
Lu LI ; Junjie WEI ; Xiaolin SUN ; Weixin WU ; Ruiqi LIU ; Haiyun LIU ; Yinze ZHANG
Chinese Journal of Blood Transfusion 2024;37(6):690-693
Objective To determine the reference red blood cells with weak agglutination intensity of low positive quali-ty control products by comparing RhD antigen expression intensity difference according to the serological results.Methods The RhD(+)red blood cells were detected by microcolumn gel method with 1 500 times diluted anti-D typing reagent.The samples with weak and strong RhD antigen expression intensity were selected as the reference red blood cells for weak agglu-tination intensity of low positive quality control products,and verification was performed.Results Ten RhD(+)red blood cells were detected with diluted anti-D typing reagent,of which 8 were 1+and 2 were±.Red blood cells with agglutination intensity of 1+were used as the benchmark to determine the maximum dilution ratio of anti-D typing reagent when their ag-glutination intensity was 1+.As the preparation standard of low positive quality control products,the agglutination intensity of red blood cells with low RhD antigen expression intensity was extremely weak±,which was difficult to ensure the stability of its control limit properties.Based on red blood cells with agglutination intensity of±,the maximum dilution ratio of anti-D typing reagent with agglutination intensity of 1+was re-determined as the preparation standard of low positive quality con-trol products,and the results met the requirements of quality control product setting.Conclusion Using red blood cells with low RhD antigen expression intensity as the benchmark to set the weak agglutination intensity of the low positive quality control products can avoid the loss of control due to the low target value.
4.Relationship between BCRP-BBB and dexmedetomidine-induced improvement in postoperative cognitive function in patients with mild hyperbilirubinemia
Yi SUN ; Haiyun WANG ; Mingshu ZHAO ; Qing LI ; Yizhen WANG
Chinese Journal of Anesthesiology 2024;44(8):927-931
Objective:To evaluate the relationship between breast cancer resistance protein (BCRP)-blood brain barrier (BBB) and dexmedetomidine-induced improvement in postoperative cognitive function in patients with mild hyperbilirubinemia.Methods:This was a prospective study. Ninety patients of both sexes with mild hyperbilirubinemia caused by choledocholithiasis, aged 55-69 yr, with body mass index of 22-28 kg/m 2, of American Society of Anesthesiologists Physical Status classification Ⅱ or Ⅲ, with preoperative Mini-Mental State Examination (MMSE) scores≥20, scheduled for elective cholecystectomy, exploratory choledocholithotomy and T-tube drainage from December 2022 to August 2023 in the Third Central Hospital of Tianjin, were divided into 2 groups( n=45 each) using a random number table method: control group (C group) and dexmedetomindine group (D group). After induction of anesthesia, dexmedetomidine was intravenously infused at a loading dose of 0.5 μg/kg over 10 min, followed by an infusion of 0.6 μg·kg -1·h -1 until the end of operation in group D, and the equal volume of normal saline was given instead in group C. At 2 days before operation and 1, 3, 5, 7 and 14 days after operation, the cognitive function was assessed using MMSE and Montreal Cognitive Assessment, and the serum BCRP concentration, concentrations of cognitive function serological indicators (serum S100β, β amyloid 42, malondialdehyde), and concentrations of BBB serological indicators (serum glial fibrillary acidic protein, thrombospondin 1, claudin 5) were determined using enzyme-linked immunosorbent assay. Results:Compared with group C, MMSE and Montreal Cognitive Assessment scores were significantly increased, the incidence of cognitive impairment was decreased, the serum concentrations of S100β, β amyloid 42 and malondialdehyde were decreased, the serum concentrations of BCRP were increased, and the serum concentrations of glial fibrillary acidic protein, claudin-5 and thrombospondin 1 were decreased in group D ( P<0.05). Conclusions:The mechanism by which dexmedetomidine improves postoperative cognitive function may be related to up-regulating BCRP levels and improving BBB function in patients with mild hyperbilirubinemia.
5.Construction of a competency evaluation indicator system for infection prevention and control nurses in Operating Rooms
Liqun SUN ; Zengmei ZHANG ; Haiyun ZHAO ; Ying ZHANG ; Zi JIN ; Yunfei NIE ; Wei WEI ; Yu WANG ; Shengyun LI ; Ruifang HUA
Chinese Journal of Modern Nursing 2024;30(13):1698-1705
Objective:To construct a competency evaluation indicator system for infection prevention and control nurses in Operating Rooms (hereinafter referred to as "IPC") and provide an objective basis for the management of IPC nurses.Methods:From June to November 2022, an initial competency evaluation indicator system for IPC nurses was developed through literature review and semi-structured interviews. The Delphi method was employed to consult 20 experts from 11 provinces and municipalities across the country. Analytical Hierarchy Process (AHP) and mean distribution method were applied to quantify and determine the weight of each level of indicators within the system.Results:Nineteen experts were finally included, with two rounds of questionnaire recovery rates of 95.00% (19/20) and 100.00% (19/19), respectively. The authority coefficients of the experts were 0.858 and 0.861, familiarity coefficients were 0.850 and 0.853, and coefficients of judgment basis were 0.865 and 0.868, respectively. The Kendall's W coefficient of concordance for the two rounds of inquiries were 0.139 and 0.202 ( P<0.05), respectively. The final IPC nurse competency evaluation indicator system included six primary indicators, 22 secondary indicators, and 66 tertiary indicators. Conclusions:The constructed IPC nurse competency evaluation indicator system is scientific, reasonable, objective, and comprehensive, providing a valuable reference for the capability training, assessment, entry standards, and qualification certification of IPC nurses.
6.Optimization of determination method for residual solvents of esmolol hydrochloride
Siming CHEN ; Xiankun WENG ; Haiyun SUN ; Zongyu LUO
Drug Standards of China 2024;25(2):154-158
Objective:To optimize the detection method for residual solvents of esmolol hydrochloride for control of internal quality.Methods:Gas chromatography headspace injection method was adopted by selecting appropriate headspace equilibrium temperature and time in order to reduce sample contamination of the instrument,optimizing solvent ratio for reducing matrix effect and adjusting carrier gas flow rate and split ratio.Polar capillary column pro-grammed temperature rise method was used with high temperature holding time,etc.,and methodological validation was conducted.Results:Ether,ethyl acetate,methanol,and toluene could all be effectively separated with good precision,and there were good linear relationships between peak areas and mass concentrations.The accuracy of sample spiking at 50%,100%,and 150%concentrations was investigated,and the average recovery rates could all reach ranges of 94.8%-105.6%.Conclusion:This method is accurate,highly sensitive,and overcomes the shortcomings of the Chinese Pharmacopoeia methods.It can be used for the detection of residual solvents in esmolol hydrochloride.
7.Changes of thickness and area of ligamenta flavum after lateral lumbar interbody fusion
Hanming BIAN ; Wentao WAN ; Chao CHEN ; Xun SUN ; Dong ZHAO ; Haiyun YANG ; Binggang GUAN ; Qiang YANG
Chinese Journal of Orthopaedics 2023;43(8):500-507
Objective:To investigate the changes of thickness and area of the ligamentum flavum after lateral lumbar interbody fusion (LLIF) for lumbar degenerative diseases.Methods:From 2019 to 2021, a total of 54 patients with lumbar degenerative diseases who underwent LLIF combined with percutaneous pedicle screw internal fixation were retrospectively analyzed. There were 9 males and 45 females, aged 59.46±6.91 years (range, 45-76 years), followed up for 14.69±6.87 months (range, 12-33 months). The disc height (DH), midsagittal canal diameter (CD), dural sac axial cross-sectional area (DCSA), ligamentum flavum area (LFA) and ligamentum flavum thickness (LFT) before and after surgery and at the last follow-up were evaluated and compared. Pearson correlation analysis was used to assess the relationship between the amount of change in the DCSA and LFA in the immediate postoperative period and at the last follow-up, as well as the correlation between the two and the amount of change in the DH. The data of patients at the last follow-up of 12 months after operation were extracted. Pearson correlation was used to evaluate the changes in DCSA and LFA at the last follow-up and the visual analogue scale (VAS) of low back pain and leg pain and Oswestry disability index (ODI) at 1 year after surgery.Results:All patients were followed up for 14.69±6.87 months (range, 12-33 months). The differences in DH ( F=354.93, P<0.001), sagittal CD ( F=44.78, P<0.001) and DCSA ( F=130.97, P<0.001) before, immediately after surgery and at the last follow-up were statistically significant. The DH, sagittal CD, and DCSA immediate after surgery and last follow-up were higher than those before surgery ( P<0.05). The differences in LFA ( F=51.59, P<0.001) and bilateral LFT ( F=53.49, P<0.001; F=50.53, P<0.001) before and after surgery and at the last follow-up were statistically significant, and both LFA and bilateral LFT at immediate after surgery and last follow-up were smaller than those before surgery ( P<0.05). Pearson correlation analysis showed that the change of DH immediately after surgery was moderately correlated with the change of DCSA ( r=0.57, P<0.001), and was strongly correlated with the change of LFA ( r=0.65, P<0.001). The change of DH at the last follow-up was moderately correlated with the change of DCSA ( r=0.43, P<0.001), and was weakly correlated with the change of LFA ( r=0.25, P=0.042). The differences in VAS-leg ( F=199.51, P<0.001), VAS-low back ( F=233.90, P<0.001), and ODI ( F=199.17, P<0.001) were statistically significant in patients before operation, 3 months after operation and 12 months after operation. There was no correlation between the changes of DCSA and LFA at the last follow-up and the changes of VAS and ODI at 1 year after operation ( P>0.05). Conclusion:LFA and LFT decrease and DCSA increase in patients with lumbar degenerative diseases after LLIF. LFA and LFT gradually decrease with time, and VAS and ODI are significantly improved compared with those before surgery. The DH loss caused by a certain degree of cage subsidence after surgery does not affect the clinical efficacy. There is no correlation between the improvement of DCSA and LFA and the improvement of clinical symptoms.
8.Role of Homer1a/mGluR5 signaling pathway in cognitive dysfunction induced by sleep deprivation in aged rats
Yun LI ; Chenyi YANG ; Xinyi WANG ; Lei FENG ; Yi SUN ; Zhuo YANG ; Haiyun WANG
Chinese Journal of Anesthesiology 2023;43(6):697-701
Objective:To evaluate the role of Homer1a/metabotropic glutamate receptor 5 (mGluR5) signaling pathway in sleep deprivation-induced cognitive dysfunction in aged rats.Methods:One hundred and four SPF healthy male Sprague-Dawley rats, aged 22-24 months, weighing 320-360 g, were divided into 4 groups ( n=26 each) using a random number table method: normal control group (group Control), sleep deprivation+ vehicle group (group SD+ Vehicle), sleep deprivation+ mGluR5 forward allosteric agent CDPPB group (group SD+ CDPPB), and sleep deprivation+ mGluR5 antagonist MPEP group (group SD+ MPEP). A 48-h sleep deprivation model was developed by sleep-deprived rod method. At the beginning of developing the model and 24 h after developing the model, CDPPB 10 mg/kg, MPEP 10 mg/kg and the equal volume of 1% Tween 80 were intraperitoneally injected in group SD+ CDPPB, group SD+ MPEP and group SD+ Vehicle, respectively.Morris water maze and novel object recognition tests were conducted to evaluate cognitive function after development of the model. The expression of Homer1a and mGluR5 in the hippocampus was detected by Western blot, the dendritic spine density in the hippocampal CA1 region was detected by Golgi staining, and the field excitatory postsynaptic potential (fEPSP) slope in the hippocampal CA1 region was detected by isolated electrophysiology. Results:Compared with Control group, the number of crossing the original platform, time of staying at the target quadrant, and novel object recognition index at 1 and 24 h after training were significantly decreased, the expression of Homer1a in the hippocampus was up-regulated, the expression of mGluR5 in the hippocampus was down-regulated, and the density of dendritic spine and fEPSP slope in the hippocampal CA1 region were decreased in group SD+ Vehicle ( P<0.05). Compared with group SD+ Vehicle, the number of crossing the original platform, time of staying at target quadrant, and novel object recognition index at 1 and 24 h after training were significantly increased, the expression of mGluR5 in hippocampus was up-regulated, and the density of dendritic spines and fEPSP slope in the hippocampal CA1 region were increased in group SD+ MPEP( P<0.05), and no statistically significant change was found in the parameters mentioned above in group SD+ CDPPB ( P>0.05). Conclusions:Sleep deprivation impairs the synaptic plasticity of hippocampal neurons by regulating Homer1a/mGluR5 signaling pathway, and thus mediating the process of cognitive dysfunction in aged rats.
9.Preparation and verification of dezocine polylactic acid-glycolic acid block copolymer microspheres
Yi SUN ; Haiyun WANG ; Mingshu ZHAO ; Xinyi WANG ; Wei HUA
Chinese Journal of Anesthesiology 2023;43(11):1378-1381
Objective:To prepare and preliminary verify dezocine polylactic acid-glycolic acid block copolymer (PLGA) microspheres.Methods:Preparation of dezocine PLGA microspheres Dezocine 120 mg, PLGA 0.1 g and the solubilizing additive poloxamer 0.1 g were dispersed in tetrahydrofuran solvent to form an organic phase solution. Sodium chloride and polyethylene glycol were dissolved in water for injection to form an inner aqueous phase solution and an outer aqueous phase solution. After the organic phase solution 20 ml was mixed with the inner aqueous phase solution 20 ml to form a water/oil colostrum, the water/oil colostrum was added to the outer aqueous phase solution to form a water/oil/water multiple emulsion, which was fully mixed with lyophilized powder protective agent and freeze-dried to prepare dezocine PLGA microspheres. Verification Eighteen clean-grade healthy male Sprague-Dawley rats, aged 10-12 weeks, weighing 220-260 g, were divided into 3 groups ( n=6 each) by a random number table method: control group (group C), dezocine ordinary preparation group (group D 1) and dezocine PLGA microspheres group (group D 2). Normal saline, dezocine injectio (dose 1 mg) and dezocine PLGA microsphere injectio (dose 0.2 μg) 0.2 ml were intramuscularly injected in C, D 1 and D 2 groups, respectively. The concentrations of dezocine in plasma were measured at 30 min and 1, 2, 3, 4, 5, 6, 7 and 8 h after administration, and thermal paw withdrawal latency was measured at T 1-T 3, T 5 and T 9. Tissues from the injection site were obtained on day 7 after intramuscular injection, and the inflammatory response was observed after HE staining. Results:Compared with group C, the thermal paw withdrawal latency was significantly prolonged at T 1-T 3 in group D 1 and at T 1-T 3, T 5 and T 9 in group D 2 ( P<0.05). Compared with group D 1, the thermal paw withdrawal latency was significantly prolonged at T 5 and T 9, and the plasma concentrations of dezocine were increased at T 6-T 9 in group D 2 ( P<0.05). Compared with the values at T 2, the plasma concentrations of dezocine were significantly decreased at T 4-T 9 in group D 1 ( P<0.05), and no significant change was found in the plasma concentrations of dezocine at T 3-T 9 in group D 2 ( P>0.05). On 7 days after injection, no inflammation was found in the local tissues in C, D 1 and D 2 groups, and no significant difference was found among the three groups. Conclusions:The sustained-release formulation of dezocine PLGA microspheres is successfully prepared, and it can maintain stable blood concentrations, effectively prolongs the action time of the drug and has significant sustained-release effect in rats.
10.Effect of Wudan Pill on Polarization of Macrophages in Endometriosis with Cold Congeal and Blood Stasis Syndrome Based on p38 MAPK Pathway
Jie ZHU ; Xiaoou XUE ; Zhiping HE ; Yong DAI ; Jun LI ; Jiawei ZHANG ; Haiyun SUN
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(24):105-113
ObjectiveTo investigate the effect of Wudan pill on the polarization of macrophages in the rat model of endometriosis (EMT) with cold congeal and blood stasis syndrome based on p38 mitogen-activated protein kinases (p38 MAPK). MethodFemale SD rats with regular motility cycles were selected and randomly divided into sham-operated group, model group, Wudan pill high, medium, and low-dose groups (2.4, 1.2, and 0.6 g⋅kg-1), Chinese patent medicine group, and western medicine group by the random number table method. The method of ice water bath + autologous endometrial transplantation was used to establish the rat model of EMT with cold congeal and blood stasis, and the rats were executed after 4 weeks of continuous drug administration to collect materials. Expression levels of serum tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-4, and transforming growth factor-β (TGF-β) were determined by enzyme-linked immunosorbent assay (ELISA) to assess inflammation. The real-time quantitative polymerase chain reaction (Real-time PCR) was performed to determine the inducible nitric oxide synthase (iNOS), TNF-α, arginase 1 (Arg1), and human mannose receptor (CD206) transcriptional levels to evaluate macrophage polarization. Western blot (WB) and immunofluorescence (IF) assays were used to determine the protein expression levels of iNOS and Arg1 to corroborate macrophage polarization. WB and Real-time PCR were used to determine the protein expression levels of the p38 MAPK pathway. ResultAs compared with sham-operated group, the levels of serum TNF-α, IL-1β, TGF-β, and IL-4 of rats in the model group were significantly higher (P<0.05). In the model group, the protein levels of iNOS, TNF-α, p-p38 MAPK, and phosphorylated-extracellular signal-regulated kinase (p-ERK) in endothelial tissues were significantly higher, the mRNA levels of iNOS, TNF-α, MAPK, and ERK were significantly higher, and the mRNA and protein expression levels of Arg1 and CD206 were significantly lower (P<0.05, P<0.01). The number of iNOS positive cells in endothelial tissues was significantly increased, and the number of Arg1 positive cells in endothelial tissues in the model group was significantly decreased (P<0.05, P<0.01). As compared with the model group, the expression of TNF-α, IL-1β, TGF-β, and IL-4 in each administration group was reduced to different degrees, which was especially significant in the Wudan pill high and medium-dose groups and the western medicine group (P<0.05). The protein expression levels of iNOS, TNF-α, p-p38 MAPK, and p-ERK in endometrial tissues of rats in the Wudan pill high and medium-dose groups, the Chinese patent medicine group, and the western medicine group were significantly lower, the mRNA expression levels of iNOS, TNF-α, MAPK, and ERK were significantly lower, and the protein expression levels of Arg1 and CD206 were significantly higher (P<0.05, P<0.01). The number of iNOS positive cells in endometrial tissues of rats was significantly decreased in the Wudan pill high and medium-dose groups, the Chinese patent medicine group, and the western medicine group, whereas the number of Arg1 positive cells was increased (P<0.05, P<0.01). The low, medium, and high doses of Wudan pill were dose-dependent, and the efficacy of the Wudan pill high-dose group was similar to that of the western medicine group. ConclusionWudan pill reduces the inflammatory response in rat model of EMT with cold congeal and blood stasis syndrome and decreases expression levels of TNF-α, IL-1β, IL-4, and TGF-β, thereby prompting polarization of macrophages from M1 to M2 type. The mechanism is presumedly related to p38 MAPK signaling pathway.

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