1.Establishment and application of a high-throughput drug screening model based on COL1A1 promoter for anti-liver fibrosis.
Shuang-Shuang ZHAO ; Ju-Xian WANG ; Yu-Cheng WANG ; Rong-Guang SHAO ; Hong-Wei HE
Acta Pharmaceutica Sinica 2015;50(2):169-173
For screening the potential drugs as anti-liver fibrosis candidates, we established a high- throughput drug screening cell model based on COL1A1 promoter. The activity of COL1A1 promoter and luciferase reporter gene can be elevated by TGF-β1, and inhibited by candidate drugs. We constructed a recombined plasmid with COL1A1 promoter and luciferase reporter gene pGL4.17, the activity of COL1A1 promoter was reflected by fluorescence intensity. COL1A1 promoter activity was detected by Dual-Luciferase Reporter Assay System, it came that the relative luciferase activity of COL1A1 promoter was 15.98 times higher than that of control group induced by TGF-β1, showing the recombined plasmid could be used in cell model. The recombined plasmid was transfected into human hepatic stellate cells LX2, detected the effect of potential drugs, and obtained a stable expression system through stable transfection and monoclonal cell culture. A sample which could reduce COL1A1 promoter activity signally by our cell model, decreased collagen I mRNA and protein expression detected by real-time RT-PCR and Western blotting. It indicates this novel cell model can be used in high-throughput drug screening of potential anti-liver fibrosis drugs.
Collagen Type I
;
genetics
;
Drug Evaluation, Preclinical
;
methods
;
Genes, Reporter
;
Hepatic Stellate Cells
;
High-Throughput Screening Assays
;
Humans
;
Liver Cirrhosis
;
drug therapy
;
Luciferases
;
Plasmids
;
Promoter Regions, Genetic
;
RNA, Messenger
;
Transfection
;
Transforming Growth Factor beta1
;
pharmacology
2.Application of WHO standards in diagnose and classification of MDS
Hong CHENG ; Wei DU ; Ming JIANG ; Jianping HAO ; Shuang CHEN ; Di ZHONG ; Yuqing MA ; Ling LI
Journal of Leukemia & Lymphoma 2012;21(5):297-299
ObjectiveTo diagnose and classify 249 patients with myelodysplastic syndrome (MDS) according to the WHO standards.MethodsAccording to the WHO standards,cell morphology,cytogenetics,immune phenotype and bone marrow pathologic biopsy in 249 cases of MDS were analyzed.ResultsGreat shape and oval cell of mature erythrocyte could be observed in all MDS patients peripheral blood. The incidence of immature erythrocyte,immature granulocyte,pelger-like abnormal nucleus and neutrophils cells without granular increased with subtypes progressing.These abnormal characteristics and proportion tended to more apparent with MDS subtypes progressing.With the dynamic follow-up,we found the rate of MDS transition to AL increased with subtypes progressing(P<0.05 ).The immune phenotype analysis of 148 patients was undertook and found that the trend to express myeloid specific antigen (CD33) increased gradually with subtypes progressing The chromosome inspection in 138 patients was undertook and found that 53 patients (38.7 % with abnormal karyotype,mainly in 20q- and +8;16 cases with complex abnormal karyotype (28 %), two patients in 5q-. 180 patients were underwent bone marrow biopsy at the same time and found that 19 patients with abnormal morphology;42 patients with bone marrow fibrosis.ConclusionsCombining with multiple index to detect the MDS contributes to the classification and diagnosis more accuratcly and long-term follow-up helps to judgment the prognosis.
3.Significance of dysplasia and cytogenetic changes on the diagnosis and typing of myelodysplastic syndrome
Ling LI ; Ming JIANG ; Hong CHENG ; Shuang CHEN ; Jianping HAO ; Di ZHONG ; Bingzhao WEN
Journal of Leukemia & Lymphoma 2012;21(7):408-411
Objective To explore the significance of dysplasia and cytogenetic changes to the diagnosis and typing of myelodysplastic syndrome (MDS).Methods The dysplasia performance of each series in every isoforms was observed by the bone marrow aspiration and peripheral blood smear to the 132 patients with MDS. At the same time do the chromosome karyotype was analizad combined with morbidness cells and chromosome karyotype abnormal analysis associated with MDS subtype. Resuits Acorrding to the dysplasia ≥0.10, the totle detection rate of granulocyte series, erythrocyte series and megakaryocytic was 43.4 %.The morbidness granulocyte and megalokaryocyte ≥0.10was mainly in RCMD (P < 0.01); morbidness erythrocytes≥0.10 mainly in RA + RARS (P < 0.01). the totle detection rate of chromosome karyotype abnormal in MDS was 44.0 %.The detection rate in RA and RARS was lower than other isoforms,but showed no statistically significant (P > 0.05).the relationships of dysplasia and chromosome karyotype abnormal with the isoforms of MDS:in RA group,50.0 %(3/6) patients had karyotype abnormal simultaneous the detection of morbidness cells≥0.10, 76.0 %(19/25) in RCMD group and 60.9 %(14/23) in RAEB group (P < 0.01).Conclusion Theve is relationships between the patients with chromosome karyotype abnormal and dysplasia ≥0.10 and the isoforms of MDS. Closely monitoring the hemopoiesis and cytogenetic changes is significance to diagnose MDS.
4.Census of pesticide residues and suggestions for control in traditional Chinese medicine.
Ying WANG ; Hong-Yu JIN ; Yan-Bin JIANG ; Lei SUN ; Shuang-Cheng MA
China Journal of Chinese Materia Medica 2014;39(5):807-811
In our study, 198 types of pesticides in 120 types 333 lots of traditional Chinese medicine (TCM), which were reasonably classified according to its matrix property, were determined by using the pretreatment platform and gas chromatography-mass spectrometry method. As a result, 158 were contaminated with pesticides. However, the content of pesticides in most TCM was very low. In addition, types of pesticides were different in different part of materia medica. In conclusion, the current status of pesticide residues pollutants in TCM was summarized, and the result can provide proof for the formulation of maximum residue limit. The new species of herbs and the new detecting index should be electively monitored in Chinese Pharmacopeia.
China
;
Drug Contamination
;
prevention & control
;
Pesticide Residues
;
analysis
;
Plants, Medicinal
;
chemistry
;
Quality Control
5.Effect of progressive muscle relaxation combined with health education on the psychological state of the maintained hemodialysis patients
Hong-Qi WANG ; Dan CHENG ; Yu-Shuang ZHAO ; Mei LIU
Chinese Journal of Modern Nursing 2011;17(11):1259-1262
Objective To evaluate the effect of progressive muscle relaxation combined with health education on the psychological state of the maintained hemodialysis patients and provide basis for the psychological intervention for them. Methods Forty-two MHD patients were randomly assigned to the intervention group and control group. Patients in the control group received the conventional nursing, and patients in the intervention group received the conventional therapy combined with nursing intervention for two times. Results After the first intervention, the various psychological testing indicators of intervention group were obviously lower than those of control group and there was statistically significant difference between two groups(P < 0. 05 ). The differences of index of physiological chemistry between two groups had no statistical significance (P > 0. 05 ). After the second intervention, the various psychological testing indicators of intervention group were further lower than those of control group and there was statistical significant difference ( P < 0. 01 or P < 0. 05 ). The Hb and Ca2 + of the intervention group were remarkably higher than control group,while PO43+ and K+ of intervention group were significantly lower than control group. Both differences had statistical significance(P < 0. 05 ). Conclusions Progressive muscle relaxation combined with health education can greatly improve the psychological and physiological status of MHD patients.
7.Study of potential protective effects of blueberry on hepatic cytochrome P450 2E1 expression in rats with immune hepatic fibrosis.
Shuang LU ; Ming-liang CHENG ; Hong LI ; Jun WU
Chinese Journal of Hepatology 2012;20(7):497-502
OBJECTIVETo study the protective effects of blueberry against rat immune hepatic fibrosis, specifically through the expression of hepatic cytochrome P450 2E1.
METHODFifty Wistar rats were randomly divided into five study groups (n = 10 each): Group A: normal control group, Group B: hepatic fibrosis model group, Group C: preventive group administered blueberry juice, Group D: preventive group administered Fu-Fang-Bie-Jia-Ruan-Gan tablet, and Group E: preventive group administered a combination of blueberry juice and Fu-Fang-Bie-Jia-Ruan-Gan tablet. The hepatic fibrosis model was established by intraperitoneal injection of porcine serum once daily for 12 weeks. Simultaneously, rats in preventive groups (Groups C-E) were perfused with blueberry juice or Fu-Fang-Bie-Jia-Ruan-Gan tablet or combinations of blueberry juice and Fu-Fang-Bie-Jia-Ruan-Gan tablet, respectively, for 12 weeks. The normal control group was perfused with saline for 12 weeks. All animals were sacrificed at the end of the 12 weeks, and serum levels of alanine aminotransferase (ALT) were measured and activities of superoxide dismutase (SOD), malondialdehyde (MDA), and hydroxyproline (Hyp) in liver homogenates were determined. Pathology of hepatic fibrosis was evaluated by hematoxylin-eosin (HE) and Masson staining. Expression of CYP2E1 was detected by real-time RT-PCR, immunohistochemical techniques, and Western blotting.
RESULTSSerum ALT levels were not significantly different in the control and treatment groups (F=4.056, P more than 0.05): A: 37.87+/-4.53 U/L, B: 49.23+/-9.81 U/L, C: 39.94+/-6.32 U/L, D: 40.50+/-5.70 U/L, and E: 38.24+/-8.43 U/L. Compared with Group B, the pathological stages of hepatic fibrosis were significantly reduced in the prevention groups (C-E) (F=95.097, P less than 0.05). Hyp and MDA in liver homogenates of groups C-E were significantly lower than those of Group B (Hyp: C: 472.68+/-44.14 mug/g, D: 416.12+/-39.38 mug/g, E: 429.51+/-55.14 mug/g vs. B: 603.16+/-68.92 mug/g, F=39.315, P less than 0.05; MDA: C: 0.83+/-0.06 nmol/mg, D: 0.96+/-0.08 nmol/mg, E: 0.85+/-0.06 nmol/mg vs. B: 1.24+/-0.15 nmol/mg, F=46.376, P less than 0.05). In contrast, SOD activities in Group C-E were significantly higher than those in Group B (C: 2.47+/-0.38 U/mg, D: 1.95+/-0.45 U/mg, E: 2.16+/-0.23 U/mg vs. B: 1.56+/-0.41 U/mg, F=25.557, P less than 0.05). Compared with Group A, the mRNA and protein expressions of CYP2E1 were increased in groups B-E, however the differences did not reach statistical significance (mRNA: F=0.897, protein: F=0.492, both P more than 0.05). The mRNA and protein expressions of CYP2E1 in groups C-E were lower than those of Group B, however the differences did not reach statistical significance (mRNA: F=0.897, protein: F=0.492, P more than 0.05).
CONCLUSIONBlueberry exhibits certain protective effects against porcine serum-induced hepatic fibrosis in rats. The expression of hepatic cytochrome P450 2E1 in rats with immune hepatic fibrosis is not significantly different from the normal rats. Blueberry has no effect on the expression of hepatic cytochrome P4502E1.
Animals ; Blueberry Plants ; Cytochrome P-450 CYP2E1 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Fruit ; Immune System Diseases ; metabolism ; Liver Cirrhosis, Experimental ; metabolism ; pathology ; Male ; Malondialdehyde ; metabolism ; Plant Extracts ; pharmacology ; Rats ; Rats, Wistar ; Superoxide Dismutase ; metabolism
8.Approach to the crystalline characteristics of ceftezole sodium.
Chang-qin HU ; Shuang-hong CHENG ; Lu LU
Acta Pharmaceutica Sinica 2002;37(4):275-279
AIMTo study the crystalline characteristics of ceftezole sodium.
METHODSCeftezole sodium crystals were obtained from different solvents. X-ray diffraction, DSC, TGA, etc were used to analyze the crytals.
RESULTSCeftezole sodium crystal was easily obtained in isopropanol-water mixture. It consists of ceftizole sodium monohydrate, which consists of type I and type II two different crystal forms. Powder X-ray diffraction patterns showed differences between type I and the type II crystal forms. Peaks at 8 degrees and 18 degrees in diffractograms of the type I, but at 9 degrees and 18.6 degrees in the type II could be observed. Water molecules in different crystal forms had different combining condition. They lost during 35-117 degrees C in the type I form, but lost during 110-160 degrees C in the type II form.
CONCLUSIONStructure of ceftizole sodium monohydrate crystal obtained in different circumstance could be some vary, which influence upon the thermal stability of the compound. The type I crystal form is more stable than the type II.
Cefazolin ; analogs & derivatives ; chemistry ; classification ; Crystallization ; Crystallography, X-Ray ; Drug Stability ; Molecular Conformation ; Molecular Structure
9.Quality assessment of sulfur-fumigated paeoniae alba radix.
Zhao WANG ; Yu-Wu CHEN ; Qiong WANG ; Lei SUN ; Wei-Yi XU ; Hong-Yu JIN ; Shuang-Cheng MA
China Journal of Chinese Materia Medica 2014;39(16):3074-3078
The samples of sulfur-fumigated Paeoniae Alba Radix acquired both by random spot check from domestic market and self-production by the research group in the laboratory were used to evaluate the effects of sulphur fumigation on the quality of Paeoniae Alba Radix by comparing sulfur-fumigated degree and character, the content of paeoniflorin and paeoniflorin sulfurous acid ester, and changes of the fingerprint. We used methods in Chinese Pharmacopeia to evaluate the character of sulfur-fumigated Paeoniae Alba Radix and determinate the content of aulfur-fumigated paeoniflorin. LC-MS method was used to analyze paeoniflorin-converted products. HPLC fingerprint methods were established to evaluate the differences on quality by similarity. Results showed that fumigated Paeoniae Alba Radix became white and its unique fragrance disappeared, along with the production of pungent sour gas. It also had a significant effect on paeoniflorin content. As sulfur smoked degree aggravated, paeoniflorin content decreased subsequently, some of which turned into paeoniflorin sulfurous acid ester, and this change was not reversible. Fingerprint also showed obvious changes. Obviously, sulfur fumigation had severe influence on the quality of Paeoniae Alba Radix, but we can control the quality of the Paeoniae Alba Radix by testing the paeoniflorin sulfurous acid ester content.
Chemistry, Pharmaceutical
;
methods
;
Drugs, Chinese Herbal
;
chemistry
;
Fumigation
;
methods
;
Paeonia
;
chemistry
;
Quality Control
;
Sulfur
;
chemistry
10.Application of HbA1C for diagnosing diabetes in Harbin
Haiqiao YU ; Weilun CHENG ; Xincai HONG ; Yuru JIA ; Siying LIU ; Wei QUAN ; Weiwei SHANG ; Yue ZHOU ; Jian LI ; Shuang YAN
Chinese Journal of Endocrinology and Metabolism 2017;33(7):562-566
Objective To evaluate the sensitivity and specificity of different HbA1C cutoff points for diabetes diagnosis in high risk outpatients in Harbin.Methods A total of 2 122 high risk outpatients(male 1 032 and female 1 090)for diabetes screening in the Fourth affiliated Hospital of Harbin Medical University from April 2013 to February 2015 were included in this study, with the average age of(49.26±13.00)year. Oral glucose tolerance tests(OGTT)were conducted and HbA1C levels were examined in these patients. The sensitivity and specificity of different HbA1C cutoff points were calculated and a receiver operator characteristic(ROC)curve was then built.Results The average level of HbA1C in these subjects was(6.45±1.72)%. The prevalence of diabetes was 41.85%. The area under ROC curve(AUC)was 0.89 with the optimal cutoff point of HbA1C 6.0% and 0.68 for the highest Yonden index. The sensitivity and specificity of HbA1C 6.0% were 84.01% and 83.67% respectively. The sensitivity and specificity of HbA1C 6.5% were 62.84% and 95.92%, respectively. The AUC of HbA1C≥6.5% was 0.732. Conclusion HbA1C works well as the diagnostic standard for diabetes in high risk outpatients of Harbin city. The cutoff point of HbA1C 6.0% is suitable for screening diabetes in high risk population, and HbA1C 6.5% is appropriate for diabetes diagnosis, with high sensitivity and specificity.