1.High Performance Liquid Chromatogram-Mass Spectrometry for Helicobacter pylori Infection-associated Differentially Expressed Proteins in Urine with Relative Molecular Mass More than 10 kDa
Huifang ZHANG ; Fanliang MENG ; Lihua HE ; Jianzhong ZHANG ; Di XIAO
Chinese Journal of Gastroenterology 2016;21(3):144-150
Background:Helicobacter pylori(Hp)is an important pathogen for peptic ulcer and gastric cancer,and is reportedly associated with a variety of extragastrointestinal diseases. However,there is no body fluid detection technique for Hp infection in clinical practice. Aims:To identify Hp infection-associated differentially expressed proteins in urine with relative molecular mass more than 10 kDa and provide potential biomarkers for diagnosis of Hp infection through body fluid detection. Methods:Midstream urine was collected from volunteers in the morning,and 13 C-urea breath test was performed to determine Hp infection. Each of 15 Hp-negative and 15 Hp-positive urine samples were mixed respectively for protein extraction. Spectra data were acquired by high performance liquid chromatogram-mass spectrometry,and label-free technology was used for relative quantitative analysis. The other 26 urine samples(15 Hp-negative and 11 Hp-positive) were used for validation by full scan. IPA software was employed for bioinformatics analysis. Results:A total of 475 urinary proteins were detected by label-free quantitative analysis and 42 differentially expressed proteins were identified. Finally,11 significantly up-regulated differentially expressed proteins were confirmed by external scanning validation. Bioinformatics analysis revealed the molecular functions,biological pathways,and related diseases of these differentially expressed proteins. Conclusions:These 11 differentially expressed proteins more than 10 kDa identified in urine might be potential biomarkers for diagnosis of Hp infection and provide molecular evidence for the correlation of Hp infection with extragastrointestinal diseases.
2.Analysis on changes of purgative biopotency in different processed products of rhubarb.
Huifang LI ; Jiabo WANG ; Yi QU ; Xiaohe XIAO
China Journal of Chinese Materia Medica 2012;37(3):302-304
OBJECTIVETo determine the difference on purgative biopotency of different processed products of rhubarb and compare rhubarb before and after preparation.
METHODThe prime biopotency of rhubarb reference substance was determined by comparing with the control substance of sennoside B using rat purgative model. Comparing with calibrated rhubarb reference substance, the bioactivity different processed products of rhubarb was determined by the 2,2',2" method.
RESULTThe purgative biopotency of crude rhubarb was 825.22 U x g(-1). The purgative biopotency of alcohol-processed rhubarb was 699.05 U x g(-1), The purgative biopotency of steamed rhubarb were 459.76 U x g(-1). Carbonized rhubarb cannot be determinate.
CONCLUSIONCrude rhubarb showed a significantly declineing purgative biopotency after being processed, alcohol-processed rhubarb showed less than crude rhubarb and followed by steamed rhubarb. Carbonized rhubarb lost almost all purgative bioactivity.
Animals ; Cathartics ; administration & dosage ; pharmacology ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Male ; Mice ; Mice, Inbred ICR ; Plant Extracts ; administration & dosage ; pharmacology ; Rheum ; chemistry ; Senna Extract ; pharmacology
3.Phosphoproteomic analysis of differential expression of AGS cellular proteins in response to Helicobanter pyiori infection
Di XIAO ; Yanyan SONG ; Fei ZHAO ; Lihua HE ; Fanliang MENG ; Huifang ZHANG ; Jianzhong ZHANG
Chinese Journal of Microbiology and Immunology 2009;29(5):449-453
Objective To research the differential expression of trace phosphorylated proteins in human gastric adenocarcinoma epithelial (AGS) cells infected by Helicobacter pylori. Methods H. pylori 26695 strain infected AGS cells 4 h and AGS cells was cultivated for 4 h as a comparison. The proteins of AGS and comparison AGS cells were extracted. Their phosphorylated proteins were enriched by metal ion af-finity adsorption enrichment techniques. After desalinated and purified the phosphorylated proteins samples were separated by 2-dimensional polyacrylamide gel electrophoresis (2-DE) technique. Computer assisted image analysis was used to analyze the differential proteomic expression. The significantly differentially ex-pressed proteins were unambiguously assigned identities by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF/TOF). Results Fifteen kinds of proteins were down-regulated, 4 kinds of new proteins were observed, 1 kind of proteins were up-regulated, 1 kind of proteins unexpression. The 21 proteins that were significantly differentially expressed , including cellular calcium ion homeostasis, transcription, interpretation, protein folding and transport, ribosomal assembly, centrosome replication, chromosome stability, cellular structure, cellular proliferation and apoptosis. Conclusion H. priori can cause a wide range change to human gastric adenocarcinoma epithelial cell protein pheshorylation. This change character has great significance to further comprehensive understanding of the pathogenesis of H. pylori.
4.An analysis of the sleep condition and its influence factors in old patients with hypertension
Lijun ZHANG ; Qiong XIONG ; Lizhong ZHANG ; Jianling ZHU ; Xinjuan LIU ; Biqun ZHONG ; Huifang XIAO
Chinese Journal of Practical Nursing 2008;24(33):4-6
Objective To investigate the sleep condition and its influence factors in old patients with hypertension. Methods A total of 100 old patients with hypertension were investigated by Pitts-burgh sleep quality index (PSQI) and serf-designed questionnaire of influence factors. Results 76 per-cents of the old patients with hypertension had sleep disorders.The total score of PSQI,sleep quality,time take to fall asleep,hours of sleep,sleep effficiency,sleep disorders,hypnotic drugs and every score of daytime dysfunction were significantly higher than domestic norm(P<0.01).Surroundings(strange circumstances or noisiness), physio-pathology (disease distress or high frequency of going to toilet during nighttime),psycho-logical situations(the fear to the disease or worrying about medical fees) and drags(use of diuretics or the adverse effects of some antihypertensive agents) were the important factors affecting sleep quality. Con-clusions Old patients with hypertension have poor sleep quality,which relates with multiple factors.Valid nursing interventions should be adopted in order to control the various factors which affect sleep quality to improve patients' sleep quality.
5.Expression and correlation of survivin, COX-2 and bcl-2 in non-Hodgkin lymphoma
Huifang XIAO ; Xiongpeng ZHU ; Yijio ZHANG ; Debin LIU ; Wenqian XU ; Chuntuan LI ; Pengliang XIN
Journal of Leukemia & Lymphoma 2011;20(10):608-611
ObjectiveTo investigate the expression of survivin,COX-2 and bcl-2 in non-Hodgkin lymphoma(NHL)and the significance and correlation between them.MethodsImmunohistochemistry MaxVision systems for survivin,COX-2 and bcl-2 were conducted on 44 NHL and 20 reactive lymphoid hyperplasia (RH).ResultsThe positive expression rates of survivin,COX-2,bcl-2 in NHL were 70.45 %(31/44),68.18 % (30/44),63.64 % (28/44),respectively,and these in RH were 40.00 % (8/20),40.00 %(8/20) and 20.00 % (4/20),respectively.There was positive correlation between the expression of COX-2 and survivin (r =0.306,P =0.043),survivin and bcl-2 protein (r =0.339,P =0.040) in NHL.ConclusionCOX-2,survivin,bcl-2 are highly expressed in NHL.To detect the expression of them has clinical value to diagnosis NHL and to estimate the malignant degree of lymphoma.There are a positive correlations between the expression of COX-2 and survivin protein,and between the expression of survivin and bcl-2 protein,which indicates that they may play a synergistic role in the occurrence and development of NHL.
6.The effects of Alpha-linolenic acid on the related key gene expressions of fatty acid synthesis in HepG2 cells
Wei LI ; Tao ZHANG ; Xianxing XIE ; Wei MENG ; Huifang LIU ; Linli XIAO ; Chunfeng LI ; Yuan LIU
Chinese Journal of Comparative Medicine 2014;(9):5-8,13
Objective To investigate the effects of different doses Alpha-Linolenic acid ( ALA ) on the expressions of fatty acid synthesis-related genes in HepG2 cells.Methods HepG2 cells were divided into two groups, normal control group (NC) and high fat group (HF) in which cells were firstly cultured for 36h in the medium contained 0.5mmol/L stearic acid.Real-time quantitative PCR was taken to detect mRNA expression of genes SREBP1C, FAS and ACC which are related to fatty synthesis.While there are significant differences in fatty synthesis, 10%, 20%, 50%, 70% and 100%ALA took the place of stearic acid, 36h later, real-time quantitative PCR and Western blotting were taken to detect mRNA and protein expression of genes related to fatty synthesis.Results SREBP1C mRNA expression of ALA substitution groups were significantly lower than the high-fat group ( P <0.001) .The FAS of 0.5 mmol/L ALA group and 0.35 mmol/L ALA group were significantly lower than the high-fat group (P <0.001).ACC genes mRNA level was not significantly different from high-fat group.SREBP1C and FAS protein expression were significantly lower than the high-fat group, but ACC showed no significant difference.Conclusions Saturated fatty acids promote hepatocyte fatty acid synthesis, ALA abates fatty acid synthesis by inhibiting FAS and SREBP1C gene expression.
7.Effect of metformin on proliferation,cell cycle and apoptosis of U937 cells
Junru LI ; Huifang LI ; Jia ZHOU ; Liyun ZHANG ; Xiao LU ; Daming ZUO ; Zhengliang CHEN
Chinese Journal of Immunology 2017;33(9):1315-1319
Objective:To study the effect of metformin on proliferation,cell cycle and apoptosis of U937 cells.Methods: U937 cells were treated with different concentrations of metformin,collected cells in 24,48 and 72 hours.Subsequently,cell proliferation was assessed by CCK-8 assay,and the cell cycle and apoptosis were analyzed by flow cytometry (FCM).The expression of Bcl-2,Bax,p-AMPK,p53 were determined by Western blot.Results: The proliferation of U937 cells was inhibited by metformin in a time-and dose-dependent manner.Metformin-treated cells were arrested at G0/G1 phase,the cell frequency at G0/G1 phase was increased in a time-and dose-dependent manner.Metformin also induced cell apoptosis in a dose-dependent manner.It showed that 20 mmol/L metformin induced cell apoptosis in a time-dependent manner.The expression of p-AMPK,p53,Bax was up-regulated while Bcl-2 expression was down-regulated after metformin treatment.Conclusion: Metformin could inhibit the U937 cell proliferation,block the cell cycle at G0/G1 phase,and induce cell apoptosis,which may partially be attribute to the up-regulation of Bax,down-regulation of Bcl-2,activation of AMPK/p53 signaling.
8.Effect of birid triple viable on peptic ulcer patients with Helicobacter pylori infection.
Canxia XU ; Lijun XIAO ; Huifang ZOU
Journal of Central South University(Medical Sciences) 2010;35(9):1000-1004
OBJECTIVE:
To observe the effect of birid triple viable on peptic ulcer patients with Helicobacter pylori(H.pylori) infection.
METHODS:
A total of 120 peptic ulcer patients with H.pylori infection was randomly divided into 2 groups. The control group was treated with the triple therapy including Esomeprazole, Amoxicillin, and Furazolidone. The treatment group was treated with the same triple therapy plus birid triple viable. The pH values of the gastric juice, bacterial culture of the gastric juice, the therapy-related side effects and the healing rate of ulcer and erosive were observed before the treatment and after a 14-day-treatment respectively. The (14)C-urea breath test((14)C-UBT) was used to evaluate the H.pylori eradication rates 4 weeks after the treatment.
RESULTS:
After the treatment, pH values of the gastric juice in both groups were significantly higher than those before the treatment (P<0.05). When pH values of the gastric juice ≥4, the positive rate of bacterial culture in the treatment group was lower than that of the control group (80.0% and 97.4% respectively, P<0.05). The rate of adverse effects in the treatment group was lower than that in the control group (11.7% and 30% respectively, P<0.05).After a 2-week treatment, the healing rate of ulcer and erosion was 80% and 68.7% in the treatment group, and 73.3% and 40.7% in the control group. The healing rate of ulcer had no significant difference in the 2 groups (P>0.05),while the healing rate of erosion in the treatment group was higher than that of the control group (P<0.05).The H.pylori eradication rates evaluated by per-protocol (PP) analysis in the treatment group was significantly higher than that of the control group (80.4% and 62.4% respectively, P<0.05).
CONCLUSION
The combined use of birid triple viable and the triple therapy of H.pylori in peptic ulcer patients with H.pylori infection can reduce the bacteria of the gastric juice and therapy-related side effects. It can increase the H.pylori eradication rate and promote the healing of erosion.
Adult
;
Amoxicillin
;
administration & dosage
;
Anti-Bacterial Agents
;
administration & dosage
;
Anti-Ulcer Agents
;
administration & dosage
;
Drug Administration Schedule
;
Drug Therapy, Combination
;
Esomeprazole
;
administration & dosage
;
Female
;
Furazolidone
;
administration & dosage
;
Helicobacter Infections
;
drug therapy
;
Helicobacter pylori
;
drug effects
;
isolation & purification
;
Humans
;
Lactobacillus acidophilus
;
physiology
;
Male
;
Peptic Ulcer
;
drug therapy
;
microbiology
9.Prediction of antigenic epitopes on HA, NA amino acid sequences of novel influenza A (H7N9) virus and analysis association between susceptibility and HLA-Ⅱalleles
Xueting LIU ; Shan WANG ; Junyan ZHANG ; Zhaoyu LIU ; Huifang CHEN ; Zehong ZOU ; Lanyan XIAO ; Zhiheng JI ; Ying HE
Chinese Journal of Immunology 2015;(1):16-21
Objective:To compare the amino acid sequences difference of HA,NA novel influenza virus A/H7N9 isolates, decipher possible B cell epitopes and T cell epitopes of HA,NA protein,and analyze the association between susceptibility and HLA polymorphisms.Methods:The amino acid sequences of novel influenza A ( H7N9) virus were downloaded from Genbank.Phylogenetic trees were constructed based on the amino acid sequences of HA and NA by using software Clustal X and MEGA 4.0.B cell and T cell epitopes were respectively predicted with Protean software and NetMHCⅡ2.2 Server online server.Results:The homology of HA and NA proteins of H7N9 virus was high.10 B cell epitopes and 15 T cell epitopes were randomly distributed throughout HA sequence and 12 B cell epitopes and 9 T cell epitopes were randomly distributed throughout NA sequence.HLA-DRB1*0701 allele which was commonly observed in Northern Chinese population have a high binding affinity for 9-mer peptides of HA and NA proteins.Conclusion:The prediction of B and T cell epitopes of HA and NA proteins with multiple methods benefits the research and development of vaccine against human infection with avian influenza A H7N9 virus.HLA-DRB1*0701 allele may contribute to susceptibility to novel influenza A (H7N9) virus.H7N9 influenza virus is more easily spread in Urumqi,Harbin,Shandong Province,Liaoning Province,Beijing, Shijiazhuang and Tianjin of China.
10.Clinical and TUBB4A mutation analysis of hypomyelination with atrophy of the basal ganglia and cerebellum in a family
Xiaolong DENG ; Huifang YAN ; Jiangxi XIAO ; Ye WU ; Qiang GU ; Haoran JI ; Dongxiao LI ; Yuwu JIANG ; Jingmin WANG
Chinese Journal of Applied Clinical Pediatrics 2016;31(24):1867-1870
Objective To analyze the clinical data and TUBB4A mutation of hypomyelination with atrophy of the basal ganglia and cerebellum (HABC)in a family,thus to provide accurate genetic counseling and prenatal diagno-sis for this family with HABC,and also to provide clinical experience for the diagnosis of HABC in China.Methods The clinical data of the proband and her family members were collected at the Department of Pediatrics,Peking Univer-sity First Hospital,December 201 4,including medical history,physical signs,and brain MRI,biochemical tests and metabolic disease screening.The associated gene of hereditary leukoencephalopathy was screened for the proband and her family members were screened by targeting -high -throughput sequencing technology,and then the genetic varia-tions were verified by Sanger sequencing.With those detection methods,the gene mutation was confirmed,and then ge-netic features were analyzed.Results Clinical features were as follows:nystagmus as the first symptom,and motor and mental retardation,dystonia and ataxia followed.Brain MRI indicated hypomyelination of white matter and atrophy of the basal ganglia and cerebellum.The clinical diagnosis of HABC was established based on the clinical features and brain MRI features above.Genetics features showed that one novel TUBB4A c.974G >T heterozygous missense muta-tion was found from the proband,which caused an amino acid change from the Trp into Leu (p.Trp325Leu).Both of her parents with normal phenotype were of wild -type in this site.Conclusions The proband from this family was diagnosed clinically based on her clinical data.One novel TUBB4Ac.974G > T (p.Trp325Leu)was founded in this study.Therefore,the spectrum of TUBB4A mutation will be expanded.In addition,this study elucidated clinical and genetic characteristics in this family with HABC,which may lay a solid foundation for the accurate genetic counseling and prenatal diagnosis.This study reported the first case of HABC caused by TUBB4A mutation in China.