1.Determination of residual solvents in amidoxime EN chelating agents by headspace gas chromatography
Jinshou ZHU ; Bingkun XIAO ; Xiaoyao MIAO ; Fang YANG ; Rongqing HUANG
Military Medical Sciences 2025;49(5):370-373
Objective To establish an analytical method for the determination of methanol and ethanol solvent residues in amidoxime EN chelating agents by headspace gas chromatography.Methods The samples were analyzed in a GC-2030 gas chromatograph system using a Shimadzu HS-20 fully automated headspace injector and quantified by an internal standard method.The samples were separated and detected by the DB-WAX capillary column with a flame ionization detector(FID).The thermostatic furnace temperature was 70 ℃ and the equilibrium time of the sample bottle was 30 min.The sample flow path temperature was 150℃ and the carrier gas was high pure nitrogen.As temperature programming,the initial column temperature of 40 ℃ lasted 9 min and increased to 200 ℃ at 40 ℃/min for 2 min.Results The peak areas of the residual solvents in EN chelator showed good linearity(R2>0.999)in the mass concentration range investigated,the detection limits of solvent methanol and ethanol were 0.25 μg/mL and 0.30 μg/mL respectively,and the limits of quantification were 1.25 μg/mL and 1.50 μg/mL,respectively.The recovery of residual solvent addition ranged from 97.10%to 102.23%,and the RSD value was less than 5.00%.Conclusion This method is sensitive enough to be used for the determination of solvent residues in amidoxime EN chelating agents.
2.Distribution characteristics and proteomic analysis of glioma-associated oncogene homolog 1 positive cells during mouse orthodontic tooth movement
Anqi LIU ; Lishu ZHANG ; Xiaoyao HUANG ; Yuan CAO ; Xinyue CAI ; Fang JIN
Chinese Journal of Stomatology 2025;60(1):61-68
Objective:To explore the distribution characteristics of glioma-associated oncogene homolog 1 (Gli1) positive cells during orthodontic tooth movement process and conduct a proteomic analysis of these cells.Methods:Forty Gli1-LacZ transgenic mice were used to establish an in vivo orthodontic tooth movement (OTM) model for labeling Gli1 positive cells in Gli1-LacZ transgenic mice (OTM group) and an unforced control group, with tooth movement distance measured using micro-CT. The spatial relationship and distribution characteristics of Gli1 positive cells and H-type vessels of CD31 and endomucin (EMCN) in periodontal tissues were detected by immunofluorescence staining. Twenty Gli1-membrane-targeted tandem dimer Tomato (mT)/membrane-targeted green fluorescent protein (mG) double-genotype mice were bred and Gli1 positive cells were sorted for proteomic sequencing after tamoxifen induction. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases were used for enrichment analysis. Results:The micro-CT three-dimensional reconstruction results showed that the mesial movement of the maxillary first molar in mice after 7 days of force application was (69±15) μm, indicating the successful establishment of the Gli1-LacZ transgenic mouse OTM model. Immunofluorescence staining showed that the blood vessels in periodontal tissue were mostly H-type vessels of CD31 and EMCN. The blood vessels in the periodontal tissues are predominantly H-type vessels positive for both CD31 and EMCN. The percentage of Gli1 positive cells in the OTM group, expressed as (54.5±13.2)%, and the relative fluorescence intensity, expressed as 2.6±0.9, were both significantly greater than those in the control group, which had a Gli1 positive cell percentage of (36.3±9.1)% ( t=3.60 , P=0.002) and a relative fluorescence intensity of 1.0±0.3 ( t=5.20, P<0.001). In contrast to the control group where only a small number of Gli1 positive cells were consistent with the distribution of H-type vessels, in the OTM group the number of Gli1 positive cells increased on the tension side were closely associated with the spatial distribution of H-type vessels. GO enrichment analysis of biological processes found that a large number of proteins in Gli1 positive cells were enriched in pathways such as angiogenesis and tissue remodeling. KEGG enrichment analysis found that related proteins were mainly enriched in pathways related to angiogenesis and Gli1, such as hypoxia-inducing factor 1 signaling pathway, vascular endothelial growth factor signaling pathway and hedgehog signaling pathway. Conclusions:The number of Gli1 positive cells increased on tension side and were closely related to H-type blood vessels in response to mechanical force during orthodontic tooth movement. This may be related to profile of inducing blood vessel formation and tissue remodeling.
3.Distribution characteristics and proteomic analysis of glioma-associated oncogene homolog 1 positive cells during mouse orthodontic tooth movement
Anqi LIU ; Lishu ZHANG ; Xiaoyao HUANG ; Yuan CAO ; Xinyue CAI ; Fang JIN
Chinese Journal of Stomatology 2025;60(1):61-68
Objective:To explore the distribution characteristics of glioma-associated oncogene homolog 1 (Gli1) positive cells during orthodontic tooth movement process and conduct a proteomic analysis of these cells.Methods:Forty Gli1-LacZ transgenic mice were used to establish an in vivo orthodontic tooth movement (OTM) model for labeling Gli1 positive cells in Gli1-LacZ transgenic mice (OTM group) and an unforced control group, with tooth movement distance measured using micro-CT. The spatial relationship and distribution characteristics of Gli1 positive cells and H-type vessels of CD31 and endomucin (EMCN) in periodontal tissues were detected by immunofluorescence staining. Twenty Gli1-membrane-targeted tandem dimer Tomato (mT)/membrane-targeted green fluorescent protein (mG) double-genotype mice were bred and Gli1 positive cells were sorted for proteomic sequencing after tamoxifen induction. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases were used for enrichment analysis. Results:The micro-CT three-dimensional reconstruction results showed that the mesial movement of the maxillary first molar in mice after 7 days of force application was (69±15) μm, indicating the successful establishment of the Gli1-LacZ transgenic mouse OTM model. Immunofluorescence staining showed that the blood vessels in periodontal tissue were mostly H-type vessels of CD31 and EMCN. The blood vessels in the periodontal tissues are predominantly H-type vessels positive for both CD31 and EMCN. The percentage of Gli1 positive cells in the OTM group, expressed as (54.5±13.2)%, and the relative fluorescence intensity, expressed as 2.6±0.9, were both significantly greater than those in the control group, which had a Gli1 positive cell percentage of (36.3±9.1)% ( t=3.60 , P=0.002) and a relative fluorescence intensity of 1.0±0.3 ( t=5.20, P<0.001). In contrast to the control group where only a small number of Gli1 positive cells were consistent with the distribution of H-type vessels, in the OTM group the number of Gli1 positive cells increased on the tension side were closely associated with the spatial distribution of H-type vessels. GO enrichment analysis of biological processes found that a large number of proteins in Gli1 positive cells were enriched in pathways such as angiogenesis and tissue remodeling. KEGG enrichment analysis found that related proteins were mainly enriched in pathways related to angiogenesis and Gli1, such as hypoxia-inducing factor 1 signaling pathway, vascular endothelial growth factor signaling pathway and hedgehog signaling pathway. Conclusions:The number of Gli1 positive cells increased on tension side and were closely related to H-type blood vessels in response to mechanical force during orthodontic tooth movement. This may be related to profile of inducing blood vessel formation and tissue remodeling.
4.Research on the model construction of medical consumables replenishment strategy and inventory control strategy based on supply-process-distribution mode
Yucong YANG ; Xiaoyao WANG ; Guangcai BAO ; Wei HUANG ; Wei LIANG
China Medical Equipment 2024;21(7):144-148
Objective:To build a medical consumables demand prediction model to predict the demand for medical consumables,and to reduce the occurrence of emergency replenishment and shortage of medical consumables.Methods:The medical consumables demand prediction model was established using support vector regression(SVR)algorithm,the replenishment strategy and inventory control strategy optimization model was established using multiple population genetic algorithm(MPGA)to build a medical consumables demand forecasting model,and the replenishment strategy and inventory control strategy were automatically generated.The relevant data of supply-processing-distribution(SPD)mode introduced by Benxi Central Hospital for medical consumables management from January 2019 to May 2023 were selected for model training,model validation test(test set)and model application prediction(application set),respectively.The average absolute prediction error,peak prediction error and trough of prediction error of the model were evaluated,and the average ratio of consumption per day to inventory cost,the average number of urgent orders per month,the average number of out of stock per month,the average number of non-urgent orders per month and the decreasing range of the index weekly average were compared.Results:There was no statistically significant difference in the average absolute prediction error between the model test set and the application set(P>0.05).The average absolute prediction error of the model was 0.033 5±0.024 5,the peak prediction error was 0.071 7 and the trough of prediction error was-0.0090.After the application of the model,the ratio of average daily consumption to inventory cost,the average number of monthly emergency order frequency,the average number of shortage of stock,and the average number of monthly non-emergency orders were(0.457 5±0.060 3),(23.95±6.04),(5.58±2.17),and(20.68±2.77),respectively.The ratio of daily average consumption to inventory cost was higher than that before the application,the average number of monthly emergency order,shortage of stock,and monthly non-emergency order were lower than those before the application,the difference was statistically significant(F=371.912,88.486,124.472,142.138,P<0.000).After the application of the model,the weekly average inventory amount decreased by 43.66%,and the average number of emergency replenishment decreased by 53.76%,the average number of shortages of stock decreased by 76.95%,and the average shortage of stock normal replenishments decreased by 34.41%.Conclusion:The medical consumables demand prediction model can predict the demand for medical consumables,optimize replenishment and inventory control strategies,reduce the cost of medical consumables inventory,reduce occurrence of emergency replenishment and shortages of stock,and reduce the number of normal replenishment.
5.Research on the Drug Renal Toxicity Biomarkers Based on HK-2 Cells
Ruiping XU ; Bingkun XIAO ; Xiaoyao MIAO ; Zhiheng LI ; Rongqing HUANG
Herald of Medicine 2024;43(8):1187-1191
Objective Screening for sensitive biomarkers for predicting and analyzing drug-induced renal toxicity,accelerates drug early development.Methods Focuses on epithelial cells of proximal convoluted tubules in human renal cortex(human kidney-2,HK-2)cells as the research object,screening for highly sensitive biomarkers using three nephrotoxic drugs(cisplatin,gentamicin,and aristolochic acid Ⅰ).Results The sensitivity of biomarkers in intracellular fluid is higher than in extracellular fluid,compared to detecting a single biomarker in the intracellular fluid.The combined detection of β2-microglobulin(β2-MG)and neutrophil gelatinase-associated lipocalin(NGAL)improved the accuracy of renal toxicity evaluation.Conclusion Based on the HK-2 cell model,the combined detection of β2-MG and NGAL in intracellular fluid can be used to predict renal toxicity in the drug's early development stage.
6.Identification and diagnostic value of characteristic volatile organic compounds in exhaled breath of mice with early stage radiation injury
Bingkun XIAO ; Xiaoyao MIAO ; Zhiheng LI ; Xiaofei ZHENG ; Rongqing HUANG
Military Medical Sciences 2024;48(2):88-94
Objective To investigate the characteristic volatile organic compounds(VOCs)in exhaled breath and their diagnostic value in mice with early stage radiation injury.Methods The thermal desorption gas chromatography-mass spectrometry(TD-GC/MS)technique was used to analyze VOCs in exhaled breath of irradiated mice by 60Coγ-ray with 800 cGy.The characteristic VOCs in the early stage of radiation injury were identified,and a diagnostic model was established.Results The 30-day survival rate of mice was 4.2%.There were significant differences in characteristic VOCs at 7 hours after radiation injury,and thirty characteristic VOCs related to early-stage radiation injury were identified.The diagnostic value of differential metabolites in mice after irradiation was evaluated via the ROC curve,and the area under the ROC curve(AUC)of a single compound exceeded 0.8.The diagnostic model was constructed by screening 9 potential biomarkers of exhalation through Fisher discriminant analysis,and its sensitivity and specificity were close to 100%.Conclusion Analysis of VOCs in exhaled breath is expected to provide a non-invasive diagnostic method for early screening and diagnosis of radiation injury.
7.Chinese Society of Allergy Guidelines for Diagnosis and Treatment of Allergic Rhinitis.
Lei CHENG ; Jianjun CHEN ; Qingling FU ; Shaoheng HE ; Huabin LI ; Zheng LIU ; Guolin TAN ; Zezhang TAO ; Dehui WANG ; Weiping WEN ; Rui XU ; Yu XU ; Qintai YANG ; Chonghua ZHANG ; Gehua ZHANG ; Ruxin ZHANG ; Yuan ZHANG ; Bing ZHOU ; Dongdong ZHU ; Luquan CHEN ; Xinyan CUI ; Yuqin DENG ; Zhiqiang GUO ; Zhenxiao HUANG ; Zizhen HUANG ; Houyong LI ; Jingyun LI ; Wenting LI ; Yanqing LI ; Lin XI ; Hongfei LOU ; Meiping LU ; Yuhui OUYANG ; Wendan SHI ; Xiaoyao TAO ; Huiqin TIAN ; Chengshuo WANG ; Min WANG ; Nan WANG ; Xiangdong WANG ; Hui XIE ; Shaoqing YU ; Renwu ZHAO ; Ming ZHENG ; Han ZHOU ; Luping ZHU ; Luo ZHANG
Allergy, Asthma & Immunology Research 2018;10(4):300-353
Allergic rhinitis (AR) is a global health problem that causes major illnesses and disabilities worldwide. Epidemiologic studies have demonstrated that the prevalence of AR has increased progressively over the last few decades in more developed countries and currently affects up to 40% of the population worldwide. Likewise, a rising trend of AR has also been observed over the last 2–3 decades in developing countries including China, with the prevalence of AR varying widely in these countries. A survey of self-reported AR over a 6-year period in the general Chinese adult population reported that the standardized prevalence of adult AR increased from 11.1% in 2005 to 17.6% in 2011. An increasing number of original articles and imporclinical trials on the epidemiology, pathophysiologic mechanisms, diagnosis, management and comorbidities of AR in Chinese subjects have been published in international peer-reviewed journals over the past 2 decades, and substantially added to our understanding of this disease as a global problem. Although guidelines for the diagnosis and treatment of AR in Chinese subjects have also been published, they have not been translated into English and therefore not generally accessible for reference to non-Chinese speaking international medical communities. Moreover, methods for the diagnosis and treatment of AR in China have not been standardized entirely and some patients are still treated according to regional preferences. Thus, the present guidelines have been developed by the Chinese Society of Allergy to be accessible to both national and international medical communities involved in the management of AR patients. These guidelines have been prepared in line with existing international guidelines to provide evidence-based recommendations for the diagnosis and management of AR in China.
Adult
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Asian Continental Ancestry Group*
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China
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Comorbidity
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Developed Countries
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Developing Countries
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Diagnosis*
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Epidemiologic Studies
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Epidemiology
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Global Health
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Humans
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Hypersensitivity*
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Prevalence
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Rhinitis, Allergic*
8.Expression of SARS-CoV nucleocapsid protein in Bac-to-Bac Baculovirus System and antigenic analysis
Li HUANG ; Zhiwu YU ; Yuxian PAN ; Liwen QIU ; Wei HAO ; Xixia DING ; Xiaoyao CHE ; Nan YU
Chinese Journal of Immunology 2014;(6):721-725
Objective:To express SARS-CoV nucleocapsid protein in Bac-to-Bac Baculovirus Expression System and analyze the antigenicity of the recombinant protein.Methods: The SARS-CoV nucleocapsid gene was amplified by PCR.The PCR product digested with BamHⅠand SalⅠrestriction endonucleases was cloned into vector pFastBac HTC of Bac-to-Bac Baculovirus expression system.Recombinant plasmid was transformed DH 10Bac cells to obtain the recombinant Bacmid DNA.Recombinant Bacmid DNA was transferred into Sf9 cells which were inducted to express the recombinant protein in High Five cells.After purified by Ni affinity chroma-tography ,the antigenicity of the recombinant protein was analyzed by Western blot and ELISA.Results:Recombinant plasmid was con-structed successfully.The recombinant protein with the relative molecular mass of 48 kD was efficiently expressed in High Five cells and purified successfully by Ni affinity chromatography.Western blot and ELISA analysis showed that the recombinant protein could be spe -cifically recognized by the monoclonal antibody to SARS-CoV N protein and immune serum from rabbits ,respectively.The recombinant protein can specifically reacted with serum from SARS patients ,not with serum from healthy persons and patients infected with hCoV-229 E and hCoV-OC43.Conclusion: SARS-CoV nucleocapsid protein has been expressed successfully in the Bac-to-Bac Baculovirus Expression System ,and obtained good antigenicity.It is preliminary deemed that it can't reacted with serum from patients infected with hCoV-229E and hCoV-OC43.

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