1.Effects of tumor-associated macrophages on the biological behavior of A375 human malignant melanoma cells
Fang YIN ; Fei WU ; Jia CHEN ; Chuguang ZHANG ; Ningjing SONG
Chinese Journal of Dermatology 2014;47(9):619-623
Objective To evaluate the effects of tumor-associated macrophages on the proliferation,invasion and migration of human cutaneous malignant melanoma cells.Methods Cultured U937 human monocytic cells at logarithmic phase were classified into three groups to be pretreated with phorbol ester for 48 hours followed by 48-hour activation by phorbol ester (M polarization),lipopolysaccharide (LPS) at 25 mg/L (M1 polarization),and interleukin (IL)-4 at 15 μg/L (M2 polarization) respectively.Then,enzyme-linked immunosorbent assay (ELISA) was performed to determine the levels of IL-12p70 and IL-10 in the supernatant of these activated cells.A375 human malignant melanoma cells were divided into four groups to be cultured alone or with M-,M1-and M2-polarized macrophages respectively.After additional culture for different durations (24,48 and 72 hours),methyl thiazolyl tetrazolium (MTT) assay was conducted to estimate the proliferative activity,and Transwell assay to evaluate the invasion and migration activity,of the A375 cells.Results The proliferation of A375 cells was accelerated by coculture with M-and M2-polarized macrophages,but inhibited by that with M1-polarized macrophages,with significant differences among the four groups in the proliferative activity at 48 and 72 hours (all P < 0.05),but not at 24 hours (P > 0.05).Invasion assay showed that the number of A375 cells that migrated through Transwell chambers was significantly larger in M2 and M groups (147.00 ± 7.92 and 113.22 ± 8.15 respectively),but smaller in the M1 group (56.44 ± 7.55),than in the control group (84.11 ± 6.07,all P < 0.05).Similarly,migration assay revealed a significant increase in the number of A375 cells that migrated through Transwell chambers in the M2 and M(p) groups (198.33 ± 8.22 and 156.00 ± 8.83 respectively),but a significant decrease in the M1 group (97.11 ± 6.75) as compared with the control group (123.89 ± 7.01,all P< 0.05).Conclusions The proliferation,invasion and migration of A375 cells can be accelerated by IL-4-activated M2-polarized macrophages,but decelerated by LPS-activated M1-polarized macrophages.Phorbol ester tends to induce monocytic cells to differentiate into M2-polarized macrophages.
2.Effects of Anxin Granules on blood lipid and ultrastructure of aorta in rabbits with dyslipidemia
Xianming FANG ; Ying YUAN ; Yunhong WU ; Xueman YIN
Journal of Integrative Medicine 2005;3(4):299-302
OBJECTIVE: To study the effects of Anxin Granules on dyslipidemia in rabbits caused by high fat plus high cholesterol diet. METHODS: Thirty-two healthy New Zealand male white rabbits were randomly divided into 4 groups: normal control group, untreated group, Zhibituo Tablet-treated group and Anxin Granule-treated group. Rabbits in the normal control group were fed with regular chow, while rabbits in the other three groups were fed with high fat plus high cholesterol diet. Zhibituo Tablets and Anxin Granules were administered to the rabbits in Zhibituo Tablet-treated group and Anxin Granule-treated group at a daily oral dose respectively. At the end of the 10th week, the levels of serum total cholesterol (TC), triglycerides (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), apolipoprotein A(1) (ApoA1) and apolipoprotein B (ApoB) were tested in each group, and the ultrastructures of the aorta were also observed by an electron microscope. RESULTS: Anxin Granules could reduce the levels of TC, TG, LDL-C and ApoB. The results observed by electron microscope showed that, as compared with the untreated group and the Zhibituo Tablet-treated group, the atherosclerosis of aorta in the Anxin Granule-treated group was lighter. And it was found that there were few lipid droplet vacuoles in cytoplasm of the endothelial cells, and various cell organs and elastic membrane were existed, but no lipid droplet vacuoles in cytoplasm of the medial smooth muscle cells. CONCLUSION: Anxin Granules can regulate the metabolism of blood lipid and inhibit the formation of atherosclerosis caused by hyperlipidemia in rabbits.
3.Application of flow-through hybrization and gene chipon human papiliomavious subtype detection in Chongqing
Fang CUI ; Yin JIA ; Jin WU ; Liping ZHANG
Chinese Journal of Primary Medicine and Pharmacy 2008;15(9):1449-1451
Objective To evaluate the application of flow-through rapid hybridization technique and gene chip (HybriMax) on human papiUomavions (HPV) subtype in Chongqing.Methods Cervical tissue samples were taken under the colpnscope form 473 females who had cervical lesion for pathological analysis.The predictive value of HybriMax in cervical abnormality was compared with pathological results,which were used as golden standard.Resuits 13 different subtypes were found and total HPV positive rate was 63.0% (284/473) Among the 17 different subtypes ,the higher positive rote HPV subtypes were HPV16 (23.7%,112/473),HPV58 (12.7% ,60/473),HPV53(7.4% ,35/473).The HPV infection rates were higher with the worse d cervical lesion(X2=77.06,P<0.01).Conclusions The most frequent subtypes of HPV infection in Chongqing cervical lesion were HPV 16,58.HybriMax was an effective method to detect HPV subtype in clinical.
4.Isolation and identification of lung cancer stem like cells from human lung adenocarci-noma cell lines
Jia YANG ; Zhihong FANG ; Jianchun WU ; Jing XU ; Xiaolin YIN ; Fanchen ZHAO ; Yan LI
Chinese Journal of Clinical Oncology 2016;(3):89-94
Objective:To isolate cell spheres containing cancer stem cells (CSCs) from lung cancer cell lines (A549 and H1299) and iden-tify their biological characteristics. Methods:By adopting the method of serum-free suspension culture, A549 cells and H1299 cells were cultured on no-adhesion plate to form tumor spheres. Clone assay, CCK-8 assay, and Transwell assay were employed to observe proliferation, self-renewal and invasion of tumor spheres. Besides, RT-PCR was performed to compare the expression levels of stem cell markers between sphere cells and adherent cells. Adherent A549 cells and A549 cell spheres were inoculated subcutaneously in nude mice and the tumor growth was assessed. Results:Isolated CSCs from A549 cells and H1299 cells in serum-free medium (SFM) without adhesion could grow as floating cell spheres. The results demonstrated that the self-renewal, proliferation and invasion of A549 and H1299 sphere cells were stronger than parent cells (P<0.05). When compared with adherent cells, the mRNA level of expres-sion of cell spheres' stem cell markers (Oct4 and Nanog) were significantly high (P<0.05) and A549 spheres had a stronger tumorigenici-ty in nude mice. Conclusion:SFM without adhesion can be a quick and easy method to construct stem cells model from human lung adenocarcinoma cell lines A549 and H1299.
5.Change of Whole Brain Degree in Primary Insomnia
Xiaofen MA ; Shaoqing ZENG ; Jin FANG ; Yunfan WU ; Shishun FU ; Kelei HUA ; Yi YIN ; Guihua JIANG
Journal of Sun Yat-sen University(Medical Sciences) 2017;38(3):390-394
[Objective] Based on the resting state functional magnetic resonance imaging to investigate the abnormal features of the functional connectivity of resting brain neural network in the patients with primary insomnia,by using voxel-wise whole-brain finctional networks analysis of degree centrality (DC) for imaging evidence of neural mechanisms underlying primary insomnia.[Methods] The resting state fMRI were performed in 59 PI patients and 47 age,education,and sex-matched normal healthy subjects.Analysis of DC map changes between the two patient groups and the control group were performed by two sample t test.(threshold at P < 0.05).[Results] Compared with the control group,the patients with PI showed significantly reduced DC value in bilateral medial frontal gyrus (MFG),bilateral anterior cingulate gyrus (ACG),and right insula;and increased DC value in right middle temporal gyrus (MTG),and left cuneus,(CUN),P < 0.05.[Conclusion]Changes of DC value occurred in some region of brain in the P[patient groups when compared with the control group.It was indicated that DC,as a novel resting-state fMRI parameter in the voxel-wise whole-brain functional networks,might be an appealing alternative approach for further study on pathologic and neuropsychological states of PI.
6.Analysis of Newborn′s Weight Investigation in Some Areas of Henan Province
xiao-shu, LI ; hu, ZHAO ; wen-juan, YIN ; ai-hong, WU ; su-fang, SHEN
Journal of Applied Clinical Pediatrics 2004;0(09):-
Objective To investigate the full-term newborn′s weight in Zhengzhou city and nearby areas around Zhengzhou in Henan province.Methods Each group newborn′s weight was divided with sex and city.We studied the regularity of full-term newborn′s weight,and examined the cause of the newborn′s weight rising.Results The average newborn′s weight in Zhengzhou was (3449.06?453.97) g,which in nearby areas around Zhengzhou was (3352.07?429.91) g.The average newborn′s weight in Zhengzhou was 86.97 g higher than other cities (P
7.Influence of exercise rehabilitation combined diet guidance on patients after heart valve replacement
Qingchun ZHANG ; Haihui YIN ; Miaomiao CAO ; Zhong LU ; Fang WANG ; Yijun WU ; Zhongya YAN
Chinese Journal of cardiovascular Rehabilitation Medicine 2017;26(1):16-20
Objective:To observe influence of exercise rehabilitation combined diet guidance on cardiac function and quality of life (QOL) in patients after heart valve replacement .Methods:A total of 120 patients after heart valve replacement were selected and randomly divided into control group (n=60 ,received routine health education ) and experiment group (n=60 ,received diet guidance and exercise rehabilitation based on routine health education ) .After discharge ,patients re‐ceived outpatient follow‐up for six months ,body weight ,cardiac function and QOL on six months after surgery were fol‐lowed up and observed in two groups ,and the results were statistically compared .Results:Compared with control group six months after discharge ,there were significant rise in left ventricular ejection fraction [ (51.75 ± 9.30)% vs .(57.73 ± 6.13)% ] andoxygenmetabolicequivalent [(7.05±1.85)METsvs.(11.34±2.18)METs],andeachitemscoreofQOL and total score of QOL [ (4.98 ± 0.40) scores vs .(6.49 ± 0.53) scores] ,P<0.05 or <0.01;and significant reduction in body weight [(67.75 ± 10.67) kg vs .(61.25 ± 10.34) kg] in experiment group ,P<0.01. Conclusion:Exercise rehabili‐tation combined diet guidance can avoid rapid increase in body weight after surgery ,promote recovery of cardiac function and improve quality of life after surgery in patients with heart valvular diseases .
8.Impact of retinol binding protein 4 polymorphism on rosiglitazone response in Chinese Type 2 diabetic patients
Fang ZHOU ; Qiong HUANG ; Xingping DAI ; Jiye YIN ; Jing WU ; Honghao ZHOU ; Zhicheng GONG ; Zhaoqian LIU
Journal of Central South University(Medical Sciences) 2011;36(10):949-957
Objective To explore the association between rs3758539G-803A and rs10882283 T-179G polymorphism of retinol binding protein 4 (RBP4) and rosiglitazone response in Chinese type 2 diabetes mellitus (T2DM) patients.Methods A total of 472 Chinese T2DM patients and 198 healthy subjects were enrolled to identify G-803A and T-179G genotypes using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP ).assay.Forty-two T2DM patients with different G-803A or T-179G genotypes were selected to undergo a 12-week rosiglitazone treatment (4 mg/d).Serum fasting plasma glucose (FPG),postprandial plasma glucose (PPG),fasting serum insulin (FINS),glycated hemoglobin (HbAlc),postprandial serum insulin ( PINS),triglyceride (TG),low-density lipoprotein-cholesterol ( LDL-c),and high-density lipoprotein-cholesterol (HDL-c) were determined before and after the rosiglitazone treatment.Results T2DM patients with RBP4 G-803A GG genotype showed lower TG and LDL-c concentrations compared with that in the GA +AA genotype subjects.T2DM patients with RBP4 T-179G TT genotype showed lower waist-to-hip ratio (WHR),FPG and FINS values compared with that in the TG + GG genotype individuals.Patients with GG genotype of RBP4 G-803A had an enhanced rosiglitazone efficacy on FPG and FINS compared with that in the GA + AA genotype group.Patients with RBP4 T179G TG + GG genotype showed an enhanced rosiglitazone efficacy on HbAlc level compared with that in the TT genotype group.Conclusion RBP4 G-803A and T-179G polymorphism might be associated with the development of T2DM and affect the therapeutic efficacy of rosignitazone in Chinese T2DM patients.
9.Primary study of chronic kidney disease with MR diffusion tensor imaging
Dandan LUO ; Lianggeng GONG ; Luo LIAN ; Jianhua YIN ; Xiangdong FANG ; Hailong WU ; Sisi YU
Chinese Journal of Radiology 2014;48(7):555-558
Objective To study the value of MR diffusion tensor imaging(DTI) in the staging of chronic kidney disease(CKD).Method MR diffusion tensor imaging was used on 26 CKD patients.All patients were diagnosed as CKD according to the criteria of clinical diagnostic.All MR images achieved diagnostic requirements.Diethylenetriamine pentaacetic acid(99mTc-DTPA) renal dynamic imaging was used to determine the unilateral renal function.CKD patients were determined as mildly renal impairment group and moderately to severely renal impairment group based on the glomerular filtration rate(GFR).Twenty healthy volunteers underwent DTI at the same time.The cortical and medullary ADC value and FA value were measured in all subjects who underwent conventional MRI and DTI.The paired Student's t test was used to compare the cortico-medullary difference of the mean ADC and FA values in all subjects.The oneway analysis of variance(ANOVA) was carried out to assess the difference among the medullary and cortical diffusion parameters(ADC,FA) of all the three groups,and LSD was used to assess multiple comparisons.The correlation of GFR and ADC value of the patients,and FA value of the patients were tested by Pearson correlation analysis.Results The cortical and medullary ADC values of 46 subjects were(2.25±0.25) × 10-3 and(2.10±0.20) × 10-3 mm2/s respectively,FA of them were(3.48±0.61) and(4.27±0.75) respectively(t =6.754,10.043 respectively; P<0.01).The cortical ADC values of volunteers,mild renal impairment group,severely renal impairment group were(2.25±0.22) ×10-3,(2.31±0.19) ×103,(2.18±0.32) ×10-3mm2/s respectively.The medullary ADC value of the three groups were(2.09±0.19) × 10-3,(2.17±0.17) × 10-3,(2.06±0.24) × 10-3mm2/s respectively(F=0.968,0.882 respectively; P>0.05).The cortical FA values of the three groups were 3.85± 0.47,3.18±0.62,3.20±0.52 respectively.The medullary FA of the three groups were 4.92±0.38,3.93±0.57,3.62 ± 0.53(F=8.725,33.705 respectively; P<0.01).There was no correlation between cortical ADC,medullary ADC,cortical FA of the patients and GFR [(30.14±15.79) ml· min-1 · 1.73 m-2](r =0.243,0.197,0.196 respectively; P>0.05).There was a positive correlation between medullary FA of the patients and GFR (r=0.556,P=0.003).Conclusion FA value has certain value in evaluating CKD early diagnosis and renal function.
10.Validation of Microbial Limit Test Methods for Thirteen Kinds of Ointments
Li FANG ; Xinyue ZHANG ; Jingjing WU ; Lingling BAI ; Hua SU ; Yin BU
China Pharmacist 2014;(9):1580-1583
Objective:To establish the microbial limit test methods for thirteen kinds of ointments. Methods:The microbial limit of 13 kinds of ointments was respectively determined by the routine method, culture medium dilution method and membrane filtration method. Results:The recovery of the tested bacteria in the samples was above 70% by the different methods. Conclusion:The micro-bial limit test methods for thirteen kinds of ointments are stablished, which may be used in the quality control.