1.Reflections on the Cell Membrane Chromatography Used in Quality Control of Traditional Chinese Medicine
Xiaofang HOU ; Sicen WANG ; Tao ZHANG ; Langchong HE
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(3):526-528
It is very difficult to comprehensively achieve the quality control of traditional Chinese medicine (TCM), because some problems such as indicator simplification and lack of correlation between efficacy and indicator are ex-isted in the common quality control of TCM. Combined with the practical experience of our research work, this paper outlined the characteristics and applications of cell membrane chromatography (CMC) and discussed that CMC method can be used for quality control of TCM.
2.Effect of isoflurane on expression of matrix metalloproteinase 2 in human tongue cancer cell
Jingsong WEI ; Xiaofang XU ; Jingli HOU ; Jie ZHANG
Chinese Journal of Anesthesiology 2013;(3):317-319
Objective To evaluate the effect of isoflurane on the expression of matrix metalloproteinase 2 (MMP-2) in human tongue cancer cell.Methods Tea8113 cells at the logarithmic growth phase were seeded into 96-well plates (200 μl/hole) with the density of 5 × 103/ml,6-well plates (5 ml/hole) with the density of 1 ×105/ml,culture dishes (5 ml/dish) with the density of 5 × 105/ml or Transwell chamber (200 μl cell suspension for upper chamber with a density of 5 × 105/ml,10 % blood serum medium 500 μl for lower chamber).The cells wererandomly divided into 3 group (n =30 each):control group (group C),2% isoflurane 2 h group (group Ⅰ1) and 2% isoflurane 4 h group (group Ⅰ2).2% isoflurane was added to the culture medium and the cells were incubated for 2 and 4 h in groups Ⅰ1 and Ⅰ2,respectively.The cell viability was detected by MTT assay.The cell apoptosis was assessed by flow cytometry.The migration and invasion of the cells were measured by cell wound scratch assay and Transwell chamber assay,respectively.The expression of MMP-2 and MMP-2 mRNA was detected by immunocytochemistry and RT-PCR,respectively.Results The cell viability and migration of cells were significantly increased,the apoptotic rate was decreased,and the expression of MMP-2 and MMP-2 mRNA was up-regulated in groups Ⅰ1 and Ⅰ2,and the invasion of the cells were increased in group Ⅰ2 as compared with group C (P < 0.05).Compared with group Ⅰ1,the cell viability and migration and invasion of the cells were significantly increased,the apoptotic rate was decreased,and the expression of MMP-2 and MMP-2 mRNA was up-regulated in group Ⅰ2 (P <0.05).Conclusion Isoflurane enhances the migration and invasion of the cancer cells by up-regulating the expression of MMP-2 in human tongue cancer Tea8113 cells.
3.Applications of HPLC/MS in the analysis of traditional Chinese medicines
Miao LI ; Xiaofang HOU ; Jie ZHANG ; Si-Cen WANG ; Qiang FU ; Lang-Chong HE ;
Journal of Pharmaceutical Analysis 2011;01(2):81-91
In China, traditional Chinese medicines (TCMs) have been used in clinical applications for thousands of years. The successful hyphenation of high-performance liquid chromatography (HPLC) and mass spectrometry (MS) has been applied widely in TCMs and biological samples analysis. Undoubtedly, HPLC/MS technique has facilitated the understanding of the treatment mechanism of TCMs. We reviewed more than 350 published papers within the last 5 years on HPLC/MS in the analysis of TCMs. The present review focused on the applications of HPLC/MS in the component analysis, metabolites analysis, and pharmacokinetics of TCMs etc. 50% of the literature is related to the component analysis of TCMs, which show that this field is the most popular type of research. In the metabolites analysis, HPLC coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry has been demonstrated to be the powerful tool for the characterization of structural features and fragmentation behavior patterns. This paper presented a brief overview of the applications of HPLC/MS in the analysis of TCMs. HPLC/MS in the fingerprint analysis is reviewed elsewhere.
4.Evaluation urinary trehalase in patients with renal proximal tubular damage
Xiaofang HAN ; Yan TAN ; Haiqin JIA ; Wei HOU ; Jing WANG ; Hong WANG ; Ruiping LIU ; Ruixia BAI
Chinese Journal of Laboratory Medicine 2012;(12):1135-1138
Objective To investigate the value of urinary trehalase (T) in patients with renal proximal tubular damage.Methods 134 patients with kidney disease (male:66 female:68 age:18-59 ; 31cases with acute renal failure,30 cases with chronic renal failure,20 cases with drug-induced renal impairment,21 cases with renal transplantation and 32 cases with nephritic syndrome) and 101 healthy controls (58 males and 43 females) were selected.Urinary T,N-acetyl-D-glucosaminidase (NAG),β2-MG,gamma-glutamyl transferase (GGT) were detected.Data were analyzed by SPSS 11.5,including nonparametric rank test,ROC analysis.Results The level of urinary trehalase in control group was normally distributed (7.1 ± 4.1) μmol/h · g Cr (0-25 μmol/h · g Cr).There was no significant difference between men and women (t =0.63,P =0.53).Urinary T levels were significant higher in all kidney disease groups than in control group (Z =6.80,5.90,5.23,6.00,8.04,P <0.01).According to ROC curve,the area of urinary T under the ROC curve (AUC) in 134 patients was 0.9,significantly different with NAG,β2-MG,GGT area (P < 0.01),the AUCs of T were 0.94,0.85,0.90,0.90,0.91 in acute and chronic renal failure group,drug-induced renal impairment group,renal transplantation group and nephritic syndrome group,respectively; Youden index were 0.85,0.65,0.77,0.66,0.72 respectively.Corresponding to the Youden index,sensitivity and specificity were 90.3% and 95.1%,73.3% and 92.1%,85.0% and 92.1%,81.0% and 85.2%,87.5% and 84.2% respectively.Conclusions The Urinary trehalase is better than other markers in the diagnosis of the proximal renal tubular damage.It was better to evaluate the proximal tubular function early in time.The diagnostic value of urinary trehalase played a key role in diagnosis,treatment and prognosis of kidney diseases.
5.Molecular mechanism of familial hypertriglyceridemia caused by lipoprotein lipase gene mutation (C310R/E396V)
Yu LUN ; Xiaofang SUN ; Ping WANG ; Jingwei CHI ; Xu HOU ; Yangang WANG
Chinese Journal of Endocrinology and Metabolism 2017;33(8):656-661
Objective The purpose of this study is to investigate the molecular mechanisms of p.C310R(c.T928C) and p.E396V(c.A1187T) lipoprotein lipase(LPL) gene mutations in vitro, which may help to construct the spectrum of LPL gene mutations and phenotype. It also can provide accurate early diagnosis for high-risk population of familial hypertriglyceridemia and provide the basis for the development of gene targeted therapy. Methods Genomic DNA was extracted from proband′s family members′ peripheral blood cells and screened by whole-exome sequencing to verify candidate gene variations. PCR products were afterwards directly sequenced again to confirm corresponding LPL variants. At the cellular level, lentiviruses containing LPL mutations were constructed and then transfected into COS-1 cells. Functional significance of the mutants was corroborated by analyzing LPL activity and mass in the cell medium and lysates via ELISA and enzyme-fluorescent method. mRNA was assayed by RT-PCR to confirm the effect on gene transcription. Results DNA sequence analysis revealed that the proband was a heterozygote for a novel c.T928C mutation in exon 6 of LPL gene, while his nephew was a compound heterozygote for the c.T928C mutation in exon 6 and a novel c.A1187T mutation in exon 8. In vitro studies, these two mutations can cause decreased activity and mass of extracellular LPL(P<0.05). Moreover, further investigation indicated that LPL C310R mutation tremendously affected post-transcriptional modification of LPL gene, whereas LPL E396V mutation dampened intracellular LPL trafficking. Conclusion Both the mutations are pathogenic by reducing the activity and mass of LPL in the plasma, which affected normal metabolism of triglycerides.
6.Study on screening potential allergenic proteins from infant milk powders based on human mast cell membrane chromatography and histamine release assays
Ping ZHANG ; Yingdi SHI ; Xiaoshuang HE ; Wei SUN ; Yanni LV ; Xiaofang HOU
Journal of Pharmaceutical Analysis 2019;9(1):55-61
Cow's milk allergy is mainly observed in infants and young children. Most allergic reactions affect the skin, followed by the gastrointestinal and respiratory systems. Conventional diagnosis is based on po-sitive allergy studies and evaluation of parameters including IgE and IgG1 levels, acute allergic skin response and anaphylactic shock reactions. We developed a cell membrane chromatographic (CMC) method based on human mast cells (HMC-1) for screening potential allergens in infant formula milk powders (IFMP). HMC-1 cell membranes were extracted and mixed with silica to prepare cell membrane chromatography columns (10 mm × 2 mm i.d., 5 mm). Under the conditions of 0.2 mL/min flow rate and 214 nm detection wavelength, human breast milk showed no retention. However, IFMP showed clear retention. The retained fractions were collected and analyzed through matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). Four major milk proteins, i.e., α-casein, β-casein, α-lactalbumin, and β-lactoglobulin A, were identified. Furthermore, these proteins and β-lacto-globulin B showed clear retention on HMC-1/CMC columns. To test the degranulation effects of the five proteins, histamine and β-hexosaminidase release assays were carried out. All five proteins induced HMC-1 cells to release histamine and β-hexosaminidase. Also, we established a reversed phase liquid chromatographic (RPLC) method for the determination of the five proteins in IFMP and the results showed that 90% proteins in IFMP were α-casein and β-casein. We concluded that cow's milk proteins may be potential allergens and caseins cause more β-casein allergic risk than other proteins. This con-clusion was consistent with other studies.
7.Determination of S-Omeprazole Sodium and the Related Substances by RP-HPLC
Xiao HU ; Xiaohai TANG ; Yongmei XIE ; Dan YANG ; Xiaofang HUO ; Ai LIU ; Jingping HOU ; Hang SONG
China Pharmacy 2005;0(22):-
OBJECTIVE:To establish an RP-HPLC method for content determination of s-omeprazole sodium and its related substances.METHODS:The separation of s-omeprazole sodium and the related substances was carried out on a Phenomenex Luna C18 column,the mobile phase consisted of methanol-0.033 mol?L-1 ammonium dihydrogen phosphate-triethylamine (58∶41.8∶0.2,adjusted to pH 7.0 by phosphate acid).The detection wavelength was 302 nm,the flow rate was 1.0 mL?min-1,the column temperature was 25 ℃,and the sample size was 20 ?L.RESULTS:The linear range of omeprazole sodium was 10~500 mg?L-1 (r=0.999 7).The average recovery rate was 100.27% (RSD=0.74%).The average content of the related substances in samples was 0.42%.CONCLUSION:This method is simple,accurate,specific and applicable for content determination of s-omeprazole sodium and its related substances.
8.Applications of HPLC/MS in the analysis of traditional Chinese medicines
Miao LI ; Xiaofang HOU ; Jie ZHANG ; Sicen WANG ; Qiang FU ; Langchong HE
Journal of Pharmaceutical Analysis 2011;01(2):81-91
In China.traditional Chinese medicines (TCMs) have been used in clinical applications for thousands of years.The successful hyphenation of high-performance liquid chromatography (HPLC) and mass spectrometry (MS) has been applied widely in TCMs and biological samples analysis.Undoubtedly.HPLC/MS technique has facilitated the understanding of the treatment mechanism of TCMs.We reviewed more than 350 published papers within the last 5 years on HPLC/MS in the analysis of TCMs.The present review focused on the applications of HPLC/MS in the component analysis,metabolites analysis,and pharmacokinetics of TCMs etc.50% of the literature is related to the component analysis of TCMs,which show that this field is the most popular type of research.In the metabolites analysis,HPLC coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry has been demonstrated to be the powerful tool for the characterization of structural features and fragmentation behavior patterns.This paper presented a brief overview of the applications of HPLC/MS in the analysis of TCMs.HPLC/MS in the fingerprint analysis is reviewed elsewhere.
9.Progress in the analysis of adulterated additives in traditional Chinese medicines and health care products
Xiaofang HOU ; Yingdi SHI ; Sicen WANG
Journal of China Pharmaceutical University 2019;50(1):33-40
In recent years, cases of illegal addition of chemical substances into the TCMs and health-care products happened regularly. Therefore, it is particularly important to develop fast, sensitive and accurate analysis methods for detection of the adulterated chemical substances. Through literature survey of relevant papers published in 2016-2017, this article summarizes the application of various analytical techniques for adulterated chemical substances to the TCMs and health-care products with useful information for the further development of new methods and technologies in this field.
10.The relationship between neuroticism and sleep quality of patients withpermanent pacemaker implantation:mediating effect of positive psychological capital
Gaorong LYU ; Xiaojing ZHAO ; Fangyan HOU ; Xiaofang XU ; Ping LI
Chinese Journal of Practical Nursing 2019;35(2):92-96
Objective To investigate the sleep quality of pacemaker implanted patients,to analyze the relationships among neuroticism,positive psychological capital and sleep quality,and to examine the potential mediating effect of positive psychological capital on the relationship between neuroticism and sleep quality. Methods Using convenience sampling, a total of 178 pacemaker implanted patients were interviewed and analyzed in two tertiary hospitals of Shandong province. They were assessed with Big Five Inventory,The Positive Psycap Questionnaireand Pittsburgh Sleep Quality Indexfor the evaluation of neuroticism,positive psychological capital and sleep quality.The Process macro was used to test the mediating effect. Results The mean score of PSQI in patients with pacemaker implantation was (8.48± 4.50), and 71.3%of them had sleep problems.The scores of neuroticism had positively correlated with the PSQI scores(r=0.31, P<0.01), the scores of positive psychological capital had negatively correlated with the PSQI scores(r=- 0.39, P<0.01).The confidence interval from Bootstrap output indicated positive psychological capital played a mediating role between neuroticism and sleep quality (95%CI:0.06-0.23), mediation effect was 0.14, the percentage of mediation effect in total effect was 43.75%. Conclusions The relation of positive psychological capital to neuroticism and sleep quality among pacemaker implanted patients are intimate.Positive psychological capital could mediate the relationship between neuroticism and sleep quality. It is suggested that nurses can improve the patient′s sleep by enhancing the intervention of positive psychological capital according to the personality characteristics of patients.