1.A methodological study of limit test of galactosamine for total hexosamine in heparin sodium
Fangxiu YANG ; Yuxin WANG ; Xiafei LIU ; Yihong LU ; Xialei FAN
Chinese Journal of Biochemical Pharmaceutics 2015;(11):165-168
Objective To establish an ion chromatography method to determine the content of galactosamine in heparin sodium sample. Methods The content of galactosamine was determined by the ratio of response value of galactosamine and glucosamine.The determination was performed on an Dionex ICS, and the separation was carried out on a Amino acids capture column (30 mm ×3 mm), Series protect column (30 mm × 3 mm)and analytical column CarboPac PA20 (150 mm ×3 mm).The mobile phase was 14 mM potassium hydroxide solution at a flow rate of 0.4 mL/min; the column tempertature was at 30℃; the injection volume was 10μL.Results Glucosamine hydrochloride had good linearity within the range of 1.013 -16.211μg/mL(Y=2.303 4X+0.824 2,r=0.998 3), the average accuracy was 92.7%, and RSD was 3.2%(n=9), the limit of detection was 0.101 3μg/mL, and the limit of quantitation was 0.337 7μg/mL.D-Galactosamine hydrochloride had good linearity within the range of 0.010 2 -0.162 5 g/mL, (Y=31.157X-0.114 4,r=0.999 3).The accuracy was 102.1%, RSD was 2.4%(n=9).The limit of detection was 0.001 0μg/mL, and the limit of quantitation was 0.003 4μg/mL.The determination of galactosamine in 3 batches of heparin sodium raw material was not detected, (0.02 ±2.1)%, (0.03 ±1.5)%, respectively, which were all lower than the limit value (1%) of United States Pharmacopeia regulation.Conclusion The method for the determination of galactosamine in total hexose amine is successfully developed , which could be used as reference for improvement of the quality standard of heparin sodium.
2.Protective effect of Zheng Chaihu Yin on paracetamol induced acute liver injury of mice
Xiafei LIU ; Xiao WU ; Fangxiu YANG ; Yuxin WANG ; Daoquan TANG ; Mei ZHANG ; Yihong LU
Chinese Journal of Pharmacology and Toxicology 2017;31(1):101-111
OBJECTIVE To explore the effect of aqueous extract of Zheng Chaihu Yin(ZCH)on paracetamol(acetaminophen,APAP)-induced hepatotoxicity. METHODS Male ICR mice were divided into three scenarios randomly:the single treatment dose of ZCH,multiple treatment or pretreatment dose of ZCH. Each scenario had a up control group and an APAP model group,while single treatment dose of ZCH group had a ZCH group at the same time. The dose of APAP and ZCH was 500 mg·kg-1 and 36 g · kg- 1,respectively. 24 h after the last administration,plasma and liver samples were prepared. Ultra- performance liquid chromatography/quadrupole- time- of- flight mass spectrometry (UPLC-Q-TOF-MS)based metabolomics profiling was used to examine changes in plasma after expo?sure to ZCH,APAP or co-exposure to ZCH and APAP. Glutamic pyruvic transaminase (GPT) and glutamic oxaloacetic transaminas (GOT) values were determined by a biochemical auto analyzer in plasma. Histopathologic changes in the liver were observed and the area was calculated after HE staining. The data were analyzed with SPSS16.0 statistical software and the results were compared with the test between the two groups to find biomarkers. Also,SIMCA software was used for partial least squares-discriminant analysis (PLS-DA) pattern recognition. RESULTS Compared to control group, APAP dosing alone caused an increase in plasma transaminases and alterations in multiple metabolic pathways. Compared to APAP group,decrease in plasma transaminases was noted when ZCH was administered after or prior to APAP. Histopathologic results showed that in the single treatment group, multiple treatment group and pretreatment group,ZCH could alleviate the liver damage induced by APAP from (32.3 ± 12.0)% to (14.2 ± 9.9)%,(8.6 ± 7.9)% to (5.2 ± 1.7)% and (32.5 ± 10.0)% to (5.2 ± 6.4)%(P<0.05). Similarly,the PLS-DA of the LC-MS data showed that the groups dosed with APAP alone were the most distinct from controls,while animals dosed with ZCH prior to or after APAP treatment were located near control group. Metabolic spectrum results showed that ZCH could restore the changes in endogenous substances including lipid metabolism,amino acid metabolism,sugar metabolism and energy metabolism induced by APAP to normal. CONCLUSION ZCH water-extraction plays major roles in the regulation of metabolism on APAP-induced liver injury. These studies demonstrate that UPLC-Q-TOF-MS-based metabolomic analysis can be sensitively and accurately predict the initiation and progres?sion of liver injury and greatly contribute to a better understanding of the hepatoprotective effects of ZCH in a clinical environment.
3.Promotion effect of FOXCUT as a microRNA sponge for miR-24-3p on progression in triple-negative breast cancer through the p38 MAPK signaling pathway.
Xiafei YU ; Fangze QIAN ; Xiaoqiang ZHANG ; Yanhui ZHU ; Gao HE ; Junzhe YANG ; Xian WU ; Yi ZHOU ; Li SHEN ; Xiaoyue SHI ; Hongfei ZHANG ; Xiao'an LIU
Chinese Medical Journal 2024;137(1):105-114
BACKGROUND:
Triple-negative breast cancer (TNBC) is a type of highly invasive breast cancer with a poor prognosis. According to new research, long noncoding RNAs (lncRNAs) play a significant role in the progression of cancer. Although the role of lncRNAs in breast cancer has been well reported, few studies have focused on TNBC. This study aimed to explore the biological function and clinical significance of forkhead box C1 promoter upstream transcript (FOXCUT) in triple-negative breast cancer.
METHODS:
Based on a bioinformatic analysis of the cancer genome atlas (TCGA) database, we detected that the lncRNA FOXCUT was overexpressed in TNBC tissues, which was further validated in an external cohort of tissues from the General Surgery Department of the First Affiliated Hospital of Nanjing Medical University. The functions of FOXCUT in proliferation, migration, and invasion were detected in vitro or in vivo. Luciferase assays and RNA immunoprecipitation (RIP) were performed to reveal that FOXCUT acted as a competitive endogenous RNA (ceRNA) for the microRNA miR-24-3p and consequently inhibited the degradation of p38.
RESULTS:
lncRNA FOXCUT was markedly highly expressed in breast cancer, which was associated with poor prognosis in some cases. Knockdown of FOXCUT significantly inhibited cancer growth and metastasis in vitro or in vivo. Mechanistically, FOXCUT competitively bounded to miR-24-3p to prevent the degradation of p38, which might act as an oncogene in breast cancer.
CONCLUSION
Collectively, this research revealed a novel FOXCUT/miR-24-3p/p38 axis that affected breast cancer progression and suggested that the lncRNA FOXCUT could be a diagnostic marker and therapeutic target for breast cancer.
Humans
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Cell Line, Tumor
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Cell Movement/genetics*
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Cell Proliferation/genetics*
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Gene Expression Regulation, Neoplastic/genetics*
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MAP Kinase Signaling System
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MicroRNAs/metabolism*
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p38 Mitogen-Activated Protein Kinases/metabolism*
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RNA, Long Noncoding/metabolism*
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Triple Negative Breast Neoplasms/pathology*
4.The epidemiological investigation and pathogenical analysis of human brucellosis in Tacheng and Kashgar of Xinjiang Uygur Autonomous Region
Qian WANG ; Bin YAN ; Zeyu CHEN ; Songsong XIE ; Hongmei ZHANG ; Xiafei LIU ; Dianqin YU ; Wureli HAZI ; Buyun CUI ; Yuanzhi WANG
Chinese Journal of Endemiology 2019;38(1):21-24
Objective To understand the infected strains and prevalence of brucellosis in occupational population in Tacheng and Kashgar regions,Xinjiang Uygur Autonomous Region.Methods In September 2015,blood samples from occupational population (including herders,semi-agricultural and semi-pastoral,veterinarians) and non-occupational population (including students and cadres) were collected to detect Brucellaspecific antibody and bacterial nucleic acids by rose bengal plate test (RBPT),serological standard tube agglutination test (SAT) and PCR methods,respectively.The positive products of PCR were sent to Shanghai Sangon Biotechnology Co.,LTD.Then the sequence results were retrieved online using the basic alignment search tool (BLAST) in GenBank web page and uploaded to NCBI (National Center for Biotechnology Information).Results A total of 546 blood samples were tested,including 300 males,aged (55-± 15) years old,and 246 females,aged (54 ± 12) years old.The positive rates were 17.58% (96/546) and 6.78% (37/546) in 546 blood samples by serological method and genetic markers targeting omp22 and omp2,respectively.The positive rates were statistically significant (x2 =29.8,P < 0.05).Additionally,based on BLAST analysis of outer membrane protein omp22 and omp2 genes,the positive products were identified as Brucella abortus,and the sequence similarity was 100.00% (253/253,863/863 bp) to Brucella abortus strain Wisconsin genome assembly,chromosome (LT651712).Conclusions Brucellosis has a high infection rate in the occupational population of some animal husbandry-based groups in Xinjiang.The infection strain is abortive species Brucella,and health education for the occupational population and prevention of brucellosis should be strengthened to reduce the infection rate.
5.Genetic Diversity of Echinococcus granulosus Genotype G1 in Xinjiang, Northwest of China
Bin YAN ; Xiafei LIU ; Junyuan WU ; Shanshan ZHAO ; Wumei YUAN ; Baoju WANG ; Hazi WURELI ; Changchun TU ; Chuangfu CHEN ; Yuanzhi WANG
The Korean Journal of Parasitology 2018;56(4):391-396
Cystic echinococcosis (CE) caused by E. granulosus is a serious helminthic zoonosis in humans, livestock and wildlife. Xinjiang is one of high endemic province for CE in China. A total of 55 sheep and cattle livers containing echinococcal cysts were collected from slaughterhouses in Changji and Yining City, northern region of Xinjiang. PCR was employed for cloning 2 gene fragments, 12S rRNA and CO1 for analysis of phylogenetic diversity of E. granulosus. The results showed that all the samples collected were identified as G1 genotype of E. granulosus. Interestingly, YL5 and CJ75 strains were the older branches compared to those strains from France, Argentina, Australia. CO1 gene fragment showed 20 new genotype haploids and 5 new genotype haplogroups (H1-H5) by the analysis of Network 5.0 software, and the YLY17 strain was identified as the most ancestral haplotype. The major haplotypes, such as CJ75 and YL5 strains, showed identical to the isolates from Middle East. The international and domestic trade of livestock might contribute to the dispersal of different haplotypes for E. granulosus evolution.
Abattoirs
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Animals
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Argentina
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Australia
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Cattle
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China
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Clone Cells
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Cloning, Organism
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Echinococcosis
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Echinococcus granulosus
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Echinococcus
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France
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Genetic Variation
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Genotype
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Haploidy
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Haplotypes
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Helminths
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Humans
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Liver
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Livestock
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Middle East
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Polymerase Chain Reaction
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Sheep
6.Advances of human oral viral vaccine development.
Shan LI ; Xiafei LIU ; Zhaojun DUAN
Chinese Journal of Biotechnology 2023;39(9):3556-3565
Development of a vaccine that can simultaneously induce effective mucosal immunity and systemic immunity is an ideal goal to prevent mucosal pathogenic infections. The digestive tract has many sites for inducing mucosal immunity, including the mouth, stomach and small intestine. An ideal oral viral vaccine can not only induce better local and distal mucosal immunity, but also produce better systemic immunity. The oral viral vaccine has also attracted much attention because of its painless vaccination, self-administration and other advantages. Due to the complexity of human digestive tract environment and mucosal immunity, only three oral attenuated live vaccines have been successfully marketed for human use. This review summarizes the characteristics of gastrointestinal mucosal immunity, the current types and research status of oral viral vaccines, and the challenges faced by oral viral vaccines, with the hope to facilitate the research and development of oral viral vaccines for human use in China.
Humans
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Viral Vaccines
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Vaccination
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Immunity, Mucosal
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Vaccines, Attenuated
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Vaccine Development
7. Identification of Q-Markers from Hedan Tablet by employing “spider-web” mode and taking compounds’ hepatotoxicity into account
Danni WANG ; Jiamin DING ; Xin CHAI ; Jing YANG ; Yuefei WANG ; Xiafei FENG ; Chunlei LIU ; Yingzi ZENG ; Wanhui ZHOU
Chinese Herbal Medicines 2022;14(4):612-621
Objective: Due to the complicated compounds and the synergistic effect of multi-compounds, the quality control and assessment of Chinese materia medica (CMM) encounters a great challenge about how to identify the key compounds, which are directly correlated with its efficacy and safety. On the guidance of study on quality marker (Q-Marker), identification of Q-Markers was performed from Hedan Tablet (HDT) by the aid of the “spider-web” mode and hepatotoxicity evaluation derived from our previous researches and literatures. Methods: By the established ultra performance liquid chromatography with photodiode array detector (UPLC-PDA) method, online UPLC-DPPH· and offline antioxidant assay, 21 candidate compounds of HDT were systematically investigated and comprehensively evaluated by the “spider-web” mode for them properties of Q-Marker based on “content-stability-activity”. In addition, the Q-Markers related with hepatotoxicity based on our previous researches and literatures were identified. Results: Salvianolic acid B (SaB), quercetin-3-O-glucuronide (Qug), isoquercitrin (IQ) and hyperoside (Hyp) were adopted as the preferable Q-Markers of HDT according to the shaded area (A) of tested compounds in “spider-web” mode. Psoralen (Ps), isopsoralen (IP), psoralenoside (PO) and isopsoralenoside (IPO) were also strongly recommended as Q-Markers closely related with safety by considering hepatotoxicity of the accumulated Ps and IP and conversion between glycoside (PO and IPO) and aglycone (Ps and IP). Conclusion: This study provided scientific evidence for quality control and assessment of HDT, and also provided a meaningful reference for application of Q-Markers in CMM.
8.Protective effect of transient receptor potential melastatin 2 inhibitor A10 on oxygen glucose deprivation/reperfusion model.
Jianguo NIU ; Xingyu LIU ; Minghua HUANG ; Xiafei YU ; Fangfang LI ; Wei YANG ; Kang MA ; Zhuoqun HUANG
Journal of Zhejiang University. Medical sciences 2021;50(1):106-112
:To investigate the effect of transient receptor potential melastatin 2 (TRPM2) inhibitor A10 on oxygen glucose deprivation/reperfusion (OGD/R) injury in SH-SY5Y cells.:Human neuroblastoma SH-SY5Y cells were subject to OGD/R injury,and then were divided into blank control group,model control group and A10 group randomly. The cell survival rate was detected by cell counting kit 8 (CCK-8); the level of cellular reactive oxygen species (ROS) was detected by reactive oxygen detection kit; the mitochondrial membrane potential was detected by tetramethylrhodamine (TMRM) method; the number of apoptotic cells was detected by TUNEL apoptosis assay kit; the protein expression level of cleaved caspase 3 was detected by Western blot.:Compared with 3,20,30,50, has lower cytotoxicity and better inhibition effect on channel activity. Compared with the model control group,ROS level was reduced,the mitochondrial membrane potential was improved,the number of apoptosis cells was reduced ,and the expression of cleaved caspase 3 was significantly reduced in the A10 group(all <0.05). : A10 can alleviate cell damage after OGD/R by inhibiting TRPM2 channel function,reducing extracellular calcium influx,reducing cell ROS levels,stabilizing mitochondrial membrane potential levels,and reducing apoptosis.
Apoptosis
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Benzeneacetamides
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Cell Survival
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Glucose
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Humans
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Oxygen/metabolism*
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Piperidones
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Reactive Oxygen Species/metabolism*
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Reperfusion
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TRPM Cation Channels