1.Progress on mechanism of IL-32 in transformation process of gastric"inflammation to cancer"
Weijian ZHANG ; Yuqi WU ; Dishu ZHOU ; Shuya SONG ; Xinxin HONG ; Yifei XU ; Tiantian CAI ; Shaoju GUO ; Huafeng PAN ; Haiwen LI
Chinese Journal of Immunology 2025;41(9):2264-2271
IL-32 is a multifunctional cytokine with both pro-inflammatory and anti-inflammatory properties.It has been proved that expression of IL-32 increases with progression of gastric mucosal diseases and severity of gastric cancer(GC),thus participating in process of gastric"inflammation-cancer"transformation.However,how IL-32 affects malignant transformation of gastric"inflamma-tion-cancer"and finally leads to adverse outcome of GC invasion and migration is still controversial.In order to better clarify regulatory effect and possible mechanism of abnormal expression of IL-32 on different histopathological stages of gastric"inflammation-cancer"transformation,and to explore new directions and breakthroughs in molecular mechanism of early truncation and treatment of gastric precancerous lesion(GPL),we searched literatures related to IL-32 in six authoritative databases at home and abroad,such as Pubmed,Web of Science and CNKI,in past 30 years.It was found that pathogenicity or protective function of IL-32 in different histo-pathological stages of gastric"inflammation-cancer"transformation depended on its different subtypes,secretory forms,surrounding cytokine environment,disease status and genetic factors.IL-32 may regulate polarization of macrophages through NF-κB,MAPK,COX2,PR3,IDO,NOD,PKCδ,FAK and STAT3,amplify or inhibit chronic inflammatory stimulation of gastric mucosa,and thus participate in process of gastric"inflammation-cancer"transformation.Our new understanding of role of IL-32 in different stages of Cor-rea cascade may contribute to development of cytokine-directed therapy,and therapy aimed at regulating different alternative splicing subtypes of IL-32 and targeting IL-32 signals can be used as a new strategy for medical treatment of GPL and GC in future.
2.Establishment of a method for determination of sialylation levels of N-glycan profiles based on ion exchange chromatography
Hongrui YIN ; Xinxin FANG ; Ying ZHANG ; Mingming XU ; Luxia ZHENG ; Hong SHAO
Drug Standards of China 2025;26(4):366-370
Objective:To establish a method for determining the sialylation levels of N-glycan profiles based on ion-exchange chromatography in glycoprotein drugs.Methods:The separation was performed in a column picking with anion-exchange resin(4.6 mm × 250 mm,5 μm,or equivalent).Mobile phase A was 20% acetonitrile solution,while mobile phase B was a 20%acetonitrile solution containing 0.1 mol·L-1 ammonium formate(pH 4.5).The flow rate was 0.60 mL·min-1.Detection was performed using a fluorescence detector with excita-tion and emission wavelengths of 330 and 420 nm,respectively.The injection volume was 5 μL.Results:Within the target loading sample range of 40% to 160%,the peak area showed a good linear relationship with protein concentration(r>0.99).The average recovery rates at three concentration levels were 106.25%,100.00%,and 106.67%,respectively.The limit of quantification was 0.03%.Conclusion:This method is suitable for detecting the sialylation levels of N-glycan profiles in various glycoprotein drugs.
3.Progress on mechanism of IL-32 in transformation process of gastric"inflammation to cancer"
Weijian ZHANG ; Yuqi WU ; Dishu ZHOU ; Shuya SONG ; Xinxin HONG ; Yifei XU ; Tiantian CAI ; Shaoju GUO ; Huafeng PAN ; Haiwen LI
Chinese Journal of Immunology 2025;41(9):2264-2271
IL-32 is a multifunctional cytokine with both pro-inflammatory and anti-inflammatory properties.It has been proved that expression of IL-32 increases with progression of gastric mucosal diseases and severity of gastric cancer(GC),thus participating in process of gastric"inflammation-cancer"transformation.However,how IL-32 affects malignant transformation of gastric"inflamma-tion-cancer"and finally leads to adverse outcome of GC invasion and migration is still controversial.In order to better clarify regulatory effect and possible mechanism of abnormal expression of IL-32 on different histopathological stages of gastric"inflammation-cancer"transformation,and to explore new directions and breakthroughs in molecular mechanism of early truncation and treatment of gastric precancerous lesion(GPL),we searched literatures related to IL-32 in six authoritative databases at home and abroad,such as Pubmed,Web of Science and CNKI,in past 30 years.It was found that pathogenicity or protective function of IL-32 in different histo-pathological stages of gastric"inflammation-cancer"transformation depended on its different subtypes,secretory forms,surrounding cytokine environment,disease status and genetic factors.IL-32 may regulate polarization of macrophages through NF-κB,MAPK,COX2,PR3,IDO,NOD,PKCδ,FAK and STAT3,amplify or inhibit chronic inflammatory stimulation of gastric mucosa,and thus participate in process of gastric"inflammation-cancer"transformation.Our new understanding of role of IL-32 in different stages of Cor-rea cascade may contribute to development of cytokine-directed therapy,and therapy aimed at regulating different alternative splicing subtypes of IL-32 and targeting IL-32 signals can be used as a new strategy for medical treatment of GPL and GC in future.
4.Establishment of a method for determination of sialylation levels of N-glycan profiles based on ion exchange chromatography
Hongrui YIN ; Xinxin FANG ; Ying ZHANG ; Mingming XU ; Luxia ZHENG ; Hong SHAO
Drug Standards of China 2025;26(4):366-370
Objective:To establish a method for determining the sialylation levels of N-glycan profiles based on ion-exchange chromatography in glycoprotein drugs.Methods:The separation was performed in a column picking with anion-exchange resin(4.6 mm × 250 mm,5 μm,or equivalent).Mobile phase A was 20% acetonitrile solution,while mobile phase B was a 20%acetonitrile solution containing 0.1 mol·L-1 ammonium formate(pH 4.5).The flow rate was 0.60 mL·min-1.Detection was performed using a fluorescence detector with excita-tion and emission wavelengths of 330 and 420 nm,respectively.The injection volume was 5 μL.Results:Within the target loading sample range of 40% to 160%,the peak area showed a good linear relationship with protein concentration(r>0.99).The average recovery rates at three concentration levels were 106.25%,100.00%,and 106.67%,respectively.The limit of quantification was 0.03%.Conclusion:This method is suitable for detecting the sialylation levels of N-glycan profiles in various glycoprotein drugs.
5.Purity Determination and Stability Analysis of Human Epidermal Growth Factor Based on National Drug Sampling and Testing
Xinxin FANG ; Hongrui YIN ; Can WANG ; Luxia ZHENG ; Hong SHAO
Herald of Medicine 2025;44(9):1385-1389
Objective To establish a high-performance liquid chromatography method to detect the purity of human epidermal growth factor(hEGF),to compare the stability of the products of different enterprises by purity results,and to provide technical support for improving quality control and unified monitoring of hEGF based on national drug sampling and testing.Methods Agilent 300SB C8 column(250 mm×4.6 mm,5 μm)was used.Mobile phase A was 7.5 mmol·L-1 sodium phosphate buffer(pH 6.8),and mobile phase B was acetonitrile,the column temperature was 30℃,the flow rate was 0.6 mL·min-1 with gradient elution,and the detector wavelength was set at 280 nm.Results The resolution between the main peak of hEGF and the adjacent impurity peak was more than 1.5,with a detection limit of 6 ng.This method has been successfully applied to determine the purity of the bulk and final products.The results showed that the purity of the final products decreased compared with the bulk.The maximum impurity that increased significantly was the deamidation impurity identified by the mass spectrometry.Conclusion Based on the typical sample of the full chain and multi-dosage form,the established method had good specificity and resolution for the preliminary identification and purity determination of hEGF bulk and final products.
6.Purity Determination and Stability Analysis of Human Epidermal Growth Factor Based on National Drug Sampling and Testing
Xinxin FANG ; Hongrui YIN ; Can WANG ; Luxia ZHENG ; Hong SHAO
Herald of Medicine 2025;44(9):1385-1389
Objective To establish a high-performance liquid chromatography method to detect the purity of human epidermal growth factor(hEGF),to compare the stability of the products of different enterprises by purity results,and to provide technical support for improving quality control and unified monitoring of hEGF based on national drug sampling and testing.Methods Agilent 300SB C8 column(250 mm×4.6 mm,5 μm)was used.Mobile phase A was 7.5 mmol·L-1 sodium phosphate buffer(pH 6.8),and mobile phase B was acetonitrile,the column temperature was 30℃,the flow rate was 0.6 mL·min-1 with gradient elution,and the detector wavelength was set at 280 nm.Results The resolution between the main peak of hEGF and the adjacent impurity peak was more than 1.5,with a detection limit of 6 ng.This method has been successfully applied to determine the purity of the bulk and final products.The results showed that the purity of the final products decreased compared with the bulk.The maximum impurity that increased significantly was the deamidation impurity identified by the mass spectrometry.Conclusion Based on the typical sample of the full chain and multi-dosage form,the established method had good specificity and resolution for the preliminary identification and purity determination of hEGF bulk and final products.
7.Research on the mechanism of dihydroactiniolide in inhibiting the proliferation of gastric cancer cells
Lijuan CHEN ; Xinxin MA ; Guangqiang GAO ; Hong TIAN ; Jiaren LIU
Practical Oncology Journal 2025;39(2):91-98
Objective The aim of this study was to investigate the inhibitory effect of dihydroactinidiolide(DHAc)on prolif-eration of gastric cancer cells and its possible mechanism.Methods Gastric cancer MKN45 cells and AGS cells were cultured and divided into the control group(culture medium without DHAc)and treatment groups with different concentrations of DHAc(50,100,200,400,600,800,and 1000 μmol/L).MTT assay and methylene blue(MB)assay were used to detect the effect of DHAc on the via-bility of MKN45 cells and AGS cells.The changes of mitochondrial membrane potential and the cell cycle distribution of MKN45 cells treated with DHAc were detected by flow cytometry,and its effects on the cell cycle of AGS cells were also analyzed.The effects of DHAc on the expression of proteins related to the cell cycle and autophagy in MKN45 cells were detected by Western blot.Results DHAc at different concentrations of 50,100,200,400,600,and 800 μmol/L showed significant inhibitory effects on proliferation of MKN45 cells.Among them,DHAc at the concentration range of 50-400 μmol/L also arrested the cell cycle of MKN45 cells at the G0/G1 phase,down-regulated the expressions of cyclin D1 and CDK4,and significantly reduced the mitochondrial membrane poten-tial(P<0.05);In the lower concentration range of 50-200 μmol/L,DHAc could induce autophagy in MKN45 cells,as manifested by the upregulation of the LC3-II/LC3-I ratio(P<0.05).In addition,different concentrations of DHAc(100,200,400,600,800,and 1000 μmol/L)could also significantly inhibit the proliferation of AGS cells.Among them,DHAc at the concentration range of 100-400 μmol/L could arrest the cell cycle of AGS cells at the G0/G1 phase(P<0.05).Conclusion DHAc can inhibit the proliferation of gastric cancer cells.The possible mechanism is that DHAc arrests the cell cycle and induces autophagy in gastric cancer cells.
8.Clinicopathological features and molecular phenotypes of pleomorphic xanthoas-trocytoma:an analysis of 79 cases
Yu ZHANG ; Weiwei FU ; Yupeng CHEN ; Hong LI ; Weiping SHI ; Jianfeng ZHOU ; Mengyi ZHUANG ; Xinxin FAN ; Sheng ZHANG ; Xingfu WANG
Chinese Journal of Clinical and Experimental Pathology 2025;41(2):221-227,232
Purpose To analyze and discuss the clinicopathological,molecular pathological characteristics,as well as diagnostic and prognostic features of pleomorphic xanthoastrocytoma(PXA)according to the new WHO classifi-cation.Methods 79 cases of PXA were collected to analyze their pathological and clinical data.Immunohistochemis-try using the EnVision method was employed to detect the expression of CD34,ATRX,Rb,Olig-2,H3K27M,H3K27me3,IDH1 R132H,BRAF VE1 and Ki67.Sanger sequencing was used to detect mutations in H3F3A and IDH1/2.Fluorescence quantitative PCR was used to detect the BRAF V600E mutation and TERT promoter region al-terations.Fluorescence in situ hybridization(FISH)was used to detect CDKN2A and EGFR alterations.The relation-ship between clinical,pathological,molecular genetics data,and prognosis was analyzed.Results The patients'ages ranged from 9 to 69 years,with an average age of 36.4 years.Most tumors were located in the temporal lobe,frontal lobe and parietal lobe.Among the 79 cases,42 were classified as grade 2 PXA and 37 as grade 3 PXA.The tumor cells exhibited pleomorphic changes,with perivascular lymphocytic sheaths and eosinophilic bodies frequently ob-served.Grade 3 PXA exhibited more mitotic figures(average of 11.8/10 HPF),and was usually accompanied by nec-rosis,focal marginal infiltration and microvascular proliferation.Immunohistochemistry and molecular characteristics revealed frequent positivity for CD34,BRAF V600E mutation(68.1%),and CDKN2A homozygous deletion(36.8%)in PXA.Some cases showed TERT gene mutation and absent Rb expression.Univariate survival analysis in-dicated that necrosis,focal marginal infiltration,and CNS WHO grade were related to overall survival,while focal infil-tration and CNS WHO grade were the independent risk factors.Conclusion The prognosis of CNS WHO grade 3 PXA is wrose than that of grade 2 PXA.Accurate diagnosis of PXA requires the combination of the morphological features,immunohistochemical staining,and multiple molecular tests.
9.Establishment and optimization of the method for related substances in myristyl alcohol
Xinxin FANG ; Chuanmei HU ; Cuixia YAN ; Hong SHAO ; Luxia ZHENG
Drug Standards of China 2025;26(2):162-166
Objective:To establish a GC method for the determination of related substances in myristyl alcohol.Methods:Use a column packed with polyethylene glycol as the stationary phase(HP-INNOWAX,0.25 mm×30 m,0.25 μm).Maintain the column temperature at 90℃,raise the temperature to 180℃by 5℃per minutes,maintain for 25 minutes.The temperatures of injection port is 270℃.The temperatures of detector is 280℃.Cal-culate the content by normalization method.Result:Lauryl alcohol,myristyl alcohol,pentadecanol,cetyl alcohol,stearyl alcohol and oleyl alcohol showed good linear relationships within the ranges of 6.2-123.8,4.2-134.2,5.2-104.2,4.5-90.3,4.8-96.0,4.8-95.2 μg·mL-1,respectively,with the correlation coefficient above 0.999 8.The limit of detection of aforementioned fatty alcohols were 0.2,0.2,0.2,0.3,0.5 and 0.5 μg·mL-1,which were more sensitive than the method in USP monotherapy Myristyl Alcohol.The content of lauryl alcohol was 0.2%-0.6%and that of cetyl alcohol was 0.1%-0.9%in 5 batches of samples.Sum of re-lated fatty alcohols and unspecified impurities was 0.7%-2.5%.Conclusion:The method established in this study has good separation effect,high specificity and high sensitivity,and solved the problem of difficult identifica-tion of related fatty alcohols caused by high noise at the baseline of the method in USP monotherapy myristyl alco-hol.It can be used for the the determination of related substances in myristyl alcohol.It also provides reference and guidance for the quality management and control of other fatty alcohols.
10.Overview of the Research on Mechanisms and Application of Essential Oil of Aromatic Chinese Medicinals in Prevention of Respiratory Infectious Disease
Wan Ling LI ; Xinxin WU ; Xiaolei LI ; Mingzhao HAO ; Fang ZHANG ; Yue ZHANG ; Haoyue LI ; Jing ZHAO
Journal of Traditional Chinese Medicine 2025;66(6):638-644
Aromatic Chinese medicinal essential oils are volatile oils extracted from aromatic Chinese herbs, which can prevent and treat respiratory infectious diseases through multiple synergistic mechanisms including pathogen inhibition, immune regulation, and inflammatory response regulation. Essential oils are primarily used externally on the body to prevent infections and alleviate symptoms through methods like inhalation, smearing, topical application, bathing, gargling or as a suppository. They can also be utilized in the environment for disinfection and air purification, through methods like diffusion, vaporization, or spraying. The external application of essential oils extracted from Chinese aromatic herbs has the advantages of convenience, quick absorption, and simultaneous influence on both the body and mind. However, there are still challenges and deficiencies in aspects such as the positioning of functions, indications, safety, and the research on the mechanism of action. It has been proposed to combine the theory of aromatic Chinese medicinals with the characteristics of essential oils, and formulate prescriptions of Chinese medicinal essential oils under the principles of traditional Chinese medicine syndrome differentiation, and prevent and treat respiratory infectious diseases efficiently, accurately, and safely, thereby expanding the clinical application of aromatic Chinese medicinals and the preventive theory of traditional Chinese medicine.

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