1.NK cells extract HLA-G1 antigen from K562-G1 cells via trogocytosis
Danping XU ; Aifen LIN ; Weihua YAN
Chinese Journal of Microbiology and Immunology 2013;(10):721-726
Objective To investigate the mechanism of acquisition of HLA-G1 antigen by NK cells.Methods K562 cells stably expressing HLA-G1 antigen (K562-G1) were constructed.K562-G1 cells, K562 cells and shed HLA-G1 were respectively co-cultured with NK-92MI cells to observe the acquisi-tion of HLA-G by NK cells.To further investigate the mechanism , NK-92MI cells with blockage HLA-G re-ceptors were further co-cultured with K562-G1 cells and HLA-G1 proteins expressing on K 562-G1 cells were blocked and then co-cultured with NK-92MI cells. Acquisition of HLA-G 1 by NK-92MI cells was analyzed by flow cytometry and fluorescence microscopy .The effects of HLA-G1 expression on the cytotoxicity of NK-92MI cell were evaluated by flow cytometry analysis based on CD 107a labeling.R esults NK-92MI cells could quickly acquire HLA-G1 from K562-G1 cells in co-culture experiments .Blockade of HLA-G1 or its re-ceptors KIR2DL4 and ILT2 with specific mAbs did not affect the acquisition of HLA-G1 by NK-92MI cells. Moreover, HLA-G1 could significantly inhibit the cytotoxicity of NK cell ( P<0.01).Conclu sion NK-92MI cells acquire HLA-G1 from K562-G1 cells via trogocytosis , which is not associated with affinity be-tween receptor and ligand , extracellular domain of HLA-G1 or passive adhesion .
2.Polyethylenimine Polymer Promotes Gene Transduction of Adenovirus in Vitro
Danping YAN ; Hong ZHANG ; Pin MA
Herald of Medicine 2017;36(8):862-864
Objective To select polyethylenimine polymer with proper molecular weight to enhance the gene expression efficiency of Adenovirus (Ad).Methods Ad was resepcitively complexed with PEI-600,PEI-1 800 and PEI 25 000 by electrostatic adsorption.Afterwards,MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay was used to investigate the cytotoxicity of the complexes with a serial concentration,followed by cell transduction assay to select the best amount of PEI with fixed dose of Ad.In the further,the physicochemical characteristics of the optimized complexes,Zeta potential,and size distribution were studied.Results MTT showed the higher the PEI moderate molecular weight was,the greater the cytotoxity was.PEI-25 000 showed a greater cytotoxicity than that of PEI-600 and PEI-1 800.As compared with PEI-600 and PEI-25 000,PEI-1 800 significanlty promoted expression of Ad gene in A549 cells.PEI turned the Zeta potential of Ad from negative to positive charge.Conclusion In the three kinds of polymer/adenovirus compounds,the cytotoxicity of the compound is positively correlated with the polymer molecular weight and concentration.The greater molecular weight the polymer as well as the higher the concentration are,the higher the cytotoxicity is.
3.Effect of angiotensin-converting enzyme inhibitors on anemia and erythropoietin requirements in hemodialysis patients
Xiaoshi ZHONG ; Danping QIN ; Xiao XIAO ; Yan LIU
Chinese Journal of Postgraduates of Medicine 2011;34(19):23-26
Objective To observe the effect of angiotensin-converting enzyme inhibitors (ACEI) on anemia and erythropoietin (EPO) requirements in maintenance hemodialysis patients. Methods Ninety maintenance hemodialysis patients with hypertension and anemia were divided into 2 groups by random digits table, observation group (45 cases, using ACEI as antihypertensive treatment), control group [45 cases,using calcium channel blocker (CCB) as antihypertensive treatment]. The follow-up period after starting ACEI or CCB therapy was one year. The hemoglobin concentration, serum EPO, EPO requirements were compared after 0, 2, 4, 6, 8, 10, 12 months' treatment. Results In response to ACEI, the mean hemoglobin value in observation group decreased progressively, reaching statistical significance after 6 months, and it had significant difference compared with that in control group [6 months: (94.21±9.20) g/Lvs. (105.55±9.16) g/L,12 months: (95.90±6.75) g/L vs. (105.81±4.45) g/L,P <0.05]. The EPO requirements experienced a progressive increase in observation group and reached statistical significance after 8 months, compared with those in control group [8 months: ( 10 090.75±1918.35) U/week vs. (7010.32±1600.15) U/week, 12 months: (11 586.39±2009.76) U/week vs. (7068.48±1615.35) U/week,P<0.05].Serum erythropoietin concentration remained stable during the study in two groups. Conclusion ACEI can worsen anemia and reduce the efficacy of EPO in maintenance hemodialysis patients.
4.Clinical significances of the CD14bright CD16bright cell subset in peripheral blood of patients with gastric cancer
Danping XU ; Linghong GAN ; Weihua YAN ; Rui ZHENG
Chinese Journal of Microbiology and Immunology 2016;36(2):138-143
Objective To study the clinical significances of CD14bright CD16bright cell subset in pe-ripheral blood of patients with gastric cancer (GC). Methods The CD14bright CD16bright cells in peripheral blood samples collected from 124 patients with gastric cancer ( GC), 130 patients with chronic gastritis (CG) and 80 normal healthy controls (HC) were measured by using flow cytometry. Differences in the CD14bright CD16bright cells between different groups were analyzed with the Mann-Whitney U test. The feasibili-ty of using CD14bright CD16bright cells as a potential biomarker for differentiating GC patients from CG was as-sessed by using the receiver operating characteristic ( ROC) curve analysis. Correlations between the CD14bright CD16bright cells and clinicopathologic parameters of GC were analyzed with multivariate correlation analysis. Results The percentages of CD14bright CD16bright cells in peripheral blood samples and in CD14bright monomuclear cells collected from the patients with GC [median: 0. 38% (0. 23% -0. 52% ) and 6. 61%(4. 23% -9. 56% )] were significantly higher than those of the CG and HC groups [ median: 0. 11%(0. 07% -0. 15% ) and 5. 08% (3. 35% -6. 42% ); median: 0. 05% (0. 03% -0. 07% ) and 5. 09%(4. 20% -7. 40% )] (P<0. 01). The area under the ROC curve for CD14bright CD16bright cells in the peripher-al blood was 0. 934 (95% CI: 0. 900-0. 968) indicating that the value of CD14bright CD16bright cells in the di-agnosis of GC was much higher than that of alpha fetoprotein (AFP), cacino-embryonic antigen (CEA) and carbohydrate antigen CA199. The area under the ROC curve for combined multi-markers by using logistic model (CD14bright CD16bright cell subset and serum tumor markers) was 0. 947 (95% CI: 0. 920-0. 973). The CD14bright CD16bright cells were closely associated with lymphocyte cells ( P < 0. 01). Conclusion The CD14bright CD16bright cells were dramatically increased in the peripheral blood of patients with gastric cancer, which could be used as a biomarker in the diagnosis of gastric cancer.
5.Structure and antibacterial activity of quaternized zirconium phosphates
Yilong PENG ; Qiuhui YANG ; Shaozao TAN ; Wenjie ZHENG ; Yan XUE ; Danping LU ; Yingbin XU
Chinese Journal of Tissue Engineering Research 2009;13(47):9305-9308
BACKGROUND: The preparation and antibacterial activity of metal ionic zirconium phosphates has been systemically investigated now, but the applications are limited owing to the discoloration or the low antibacterial activity. Here we prepared new antibacterial agents of quaternized zirconium phosphates by introducing quaternary ammonium salt bactericidal agent with high-effective, broad-spectrum and low-toxic into sodium zirconium phosphate through an ion-exchange method.OBJECTIVE: To explore the component structure and antibacterial activity of quaternized zirconium phosphates.DESIGN, TIME AND SETTING: An in vitro observational experiment was performed at Research Laboratory of Department of Chemistry, Jinan University from June to August 2009.MATERIALS: Quaternized zirconium phosphates were prepared by introducing dodecyl dimethyl benzyl ammonium chloride into sodium zirconium phosphate through an ion-exchange method.METHODS: The mol ratios of quaternary ammonium cations to cation exchange capacity of sodium zirconium phosphate in reaction solutions were 0.25: 1,0.5: 1, 1.0: 1, and 1.5 : 1, respectively, and four kinds of quaternized zirconium phosphates containing different contents of quaternary ammonium cations (QZrP-1, QZrP-2, QZrP-3, QZrP-4) were prepared through an ion-exchange method.MAIN OUTCOME MEASURES: The component structure and heat resistance of samples were measured by using an IR spectrometer, an elemental analyzer and a thermal analyzer, respectively. The minimum inhibitory concentrations (MICs) and minimal bactericidal concentrations (MICs) of the samples against Escherichia coli (E. co/i) and Staphylococci aureus (S. aureus) were estimated by a tube broth method.RESULTS: Quaternized zirconium phosphates were prepared, and the quaternary ammonium cation content increased with increasing the concentration of quaternary ammonium cations in reaction solution. The mass fraction of quaternary ammonium cations of QZrP-1, QZrP-2, QZrP-3, and QZrP-4 was 3.70%, 5.00%, 6.96%, and 10.01%, respectively. The onset temperatures of the decomposition for quaternary ammonium cations in quaternized zirconium phosphates were all higher than 345 °C, and they were preferable thermal stability. The antibacterial activity was higher when the quaternary ammonium cation content of quaternized zirconium phosphates increased. For quaternized zirconium phosphates QZrP-3 containing 6.96% mass fraction of quaternary ammonium cations, showed excellent antibacterial activity against E. coli and S. aureus.CONCLUSION: Quaternized zirconium phosphates QZrP-3 containing 6.96% mass fraction of quaternary ammonium cations,exhibited excellent thermal stability and antibacterial activity.
6.Application of contrast-enhanced ultrasound in preoperative diagnosis of anal fistula
Dazhong ZOU ; Yifeng YU ; Danping YAN ; Li LIU ; Qin XUE ; Yufang XU ; Yuying TANG ; Hai GONG
Chinese Journal of Ultrasonography 2010;19(12):1051-1053
Objective To evaluate the clinical value of contrast-enhanced ultrasound (CEUS) in preoperative diagnosis of anal fistula. Methods Forty-five patients with fistula in ano were evaluated by physical examination, then CEUS were peformed by injecting SonoVue through the external opening to enhance the detection of the fistulous track and the internal opening. The results of CEUS were matched with surgical features to establish their accuracy in preoperative assessment of anal fistula. Results Simple typing fistula was found in 19 of 45 patients and 26 patients had complex fistulas. The accurate diagnostic rates by conventional ultrasound of simple typing fistula and complex fistulas were 89. 5% and 61.5%,respectively,the accurate diagnostic rates by CEUS of simple typing fistula and complex fistulas were 94. 7% and 92.3%, respectively, there was no significant difference between conventional ultrasound and CEUS about simple typing fistula( P >0.05), and there was significant difference about complex fistulas ( P <0. 05). Conclusions CEUS has a good visibility and accurate rate for diagnosis of anal fistula,It plays an important role for operation.
7.Analysis of risk factors related with hyperuricemia among 1420 youth and middle-aged adult health check-up residents in Guangzhou
Xuan XIONG ; Xiaoshi ZHONG ; Xiao XIAO ; Danping QING ; Jianguang HU ; Ying ZHANG ; Yan LIU
Chinese Journal of General Practitioners 2013;(7):560-562
To survey the serum uric acid (SUA) levels and associated risk factors of hyperuricemia among youth and middle-aged residents in Guangzhou.A total of 1420 subjects,aged from 20 to 60 years,receiving health check-up at our hospital in 2010 were enrolled.The total prevalence of hyperuricemia was 22.04%,32.01% in males and 14.07% in females.The average SUA was (388 ±78) μmol/L in males and (288 ± 63) μ mol/L in females.The prevalence of hyperuricemia in males was 30.11% before 40 years of age and 33.81% between 40 and 60 years of age.The average level of SUA in males was significantly higher than that of females.logistic regression analysis showed that BUN,body mass index (BMI) and hypertriglyceridemia were the independent risk factors of disease while HDL-C and gender (females) the protective factors.
8.Clinical significance of HLA-G in patients with gastric cancer
Danping XU ; Tongtong ZHANG ; Qing WANG ; Weiwu SHI ; Rui ZHENG ; Weihua YAN
Chinese Journal of Microbiology and Immunology 2016;36(7):487-493
Objective To investigate the clinical significance of CD14+HLA-G+ monocytes in pe-ripheral blood and the soluble form of HLA-G ( sHLA-G ) in plasma among patients with gastric cancer ( GC) . Methods Blood samples were collected from 135 patients with gastric cancer ( GC group) , 150 pa-tients with chronic gastritis ( CG group) and 80 healthy controls ( HC group) . Flow cytometry analysis and ELISA were used to detect the percentages of CD14+HLA-G+ monocytes in peripheral blood samples, the concentrations of sHLA-G in plasma samples and the levels of alpha fetoprotein (AFP), cacino-embryonic antigen ( CEA) , CA19-9 and CA125 in serum samples. Mann-Whitney U test was performed to analyze the differences between different groups. The feasibility of using CD14+HLA-G+ monocytes, sHLA-G, AFP, CEA, CA19-9 and CA125 as potential biomarkers to differentiate patients with GC from those with CG or healthy subjects was assessed by using receiver operating characteristic ( ROC ) curve analysis. Results The median percentages of CD14+HLA-G+ monocytes in subjects from GC, CG and HC groups were 18. 6% (12. 1%-26. 7%), 7. 3% (4. 2%-11. 0%) and 4. 6% (3. 6%-6. 3%), respectively. The percentages of CD14+HLA-G+monocytes in the peripheral blood of patients with GC were significantly higher than those in patients with CG and healthy subjects (P<0. 001). The concentrations of sHLA-G in plasma samples collected from patients with GC [(100. 6±61. 3) U/ml) were significantly higher than those in pa-tients with CG [(59.5±19. 9) U/ml) and healthy subjects [(45. 8±23. 3) U/ml] (P<0. 001). ROC curve analysis showed that in terms of GC diagnosis, the area under ROC curve ( AUC) , cutoff value, sensi-tivity and specificity for CD14+HLA-G+monocytes and sHLA-G in plasma were 0. 893 and 0. 720, 12% and 85 U/mL, 75. 8% and 50. 5%, 86. 7% and 95. 9% (P<0. 001), respectively, which indicated that CD14+HLA-G+ monocytes and sHLA-G were better than AFP, CEA, CA19-9 and CA125 in differentiating GC from CG and HC. Moreover, the multivariate logistic regression analysis revealed that the CD14+HLA-G+ monocytes, sHLA-G in plasma as well as CA19-9 and CA125 in serum were positively correlated with the risk of GC after excluding the differences caused by age and gender factors. Conclusion The levels of CD14+HLA-G+ monocytes in peripheral blood and sHLA-G in plasma increased dramatically in patients with gastric cancer, which suggested that CD14+HLA-G+monocytes and sHLA-G might be risk factors for GC and could be used as potential biomarkers for the diagnosis of GC.
9.HLA-G 14 bp insertion/deletion polymorphism is a prognostic factor for non-small-cell lung cancer
Danping XU ; Linghong GAN ; Jiamei LIU ; Dongqing LYU ; Jian LIN ; Weihua YAN
Chinese Journal of Microbiology and Immunology 2017;37(5):361-368
Objective To study the correlation between human leucocyte antigen-G (HLA-G) 14 bp insertion/deletion (I/D) polymorphism and susceptibility to non-small cell lung cancer (NSCLC) as well as poor prognosis in NSCLC.Methods A total of 113 patients with NSCLC and 150 age-and sex-matched healthy subjects were genotyped by PCR to analyze the HLA-G 14 bp insertion/deletion polymorphism in them.Epidermal growth factor receptor (EGFR) gene mutation in patients with NSCLC was detected by using amplification refractory mutation system (AMRS).Expression of HLA-G in NSCLC tissues was detected with immunohistochemistry.All patients with NSCLS were followed up to collect survival data, which were further analyzed with Kaplan-Meier method.Results The frequency of HLA-G 14 bp D/D genotype was significantly higher in the patients with NSCLC than that in the healthy subjects (x2=3.907, P=0.048, OR=1.66).Among the patients with NSCLC, HLA-G 14 bp I/I genotype carriers had a shorter overall survival time as compared with that of HLA-G 14 bp I/D or HLA-G 14 bp D/D genotype carriers (P=0.005).Patients who received chemotherapy or radiation had significantly shorter survival time than those received EGFR-targeted therapy (P=0.001).Among patients who were positive for EGFR mutation, HLA-G 14 bp D/D genotype carriers had longer survival time than those carrying HLA-G 14 bp I/I or HLA-G 14 bp I/D genotype (P=0.041).The expression of HLA-G was closely correlated with HLA-G 14 bp polymorphism in patients with NSCLC (P=0.001).Conclusion These data, reported for the first time, indicates that HLA-G 14 bp polymorphism might be a genetic factor related to the susceptibility to NSCLC and associated with survival in patient with NSCLC after excluding the interference of molecular targeted agents.
10.Clinical research of transgastric approach transluminal endoscopic surgery in diagnosing unexplained ascites
Jing TANG ; Dan YANG ; Tao WU ; Wenyong SHEN ; Danping LU ; Sha WEI ; Yan XUE ; Meihua YANG ; Aimin LIU
Chongqing Medicine 2017;46(12):1619-1620,1623
Objective To explore the clinical value of transgastric natural orifice transluminal endoscopic surgery(NOTES)in diagnosing unexplained ascites.Methods The clinical data in 12 cases of unexplained ascites diagnosed by adopting transgastric approach NOTES and performed abdominal exploration and peritoneal biopsy in our hospital from November 2015 to July 2016 were retrospectively analyzed.The operative risk and clinical application value were evaluated by statistically analyzing the postoperative complications occurrence and the diagnosis rate of disease.Results The definite diagnosis rate reached 100% verified by pathology after abdominal exploration and peritoneal biopsy,in which 8 cases(66.7%)were tuberculous peritonitis,2 cases(16.7%)were liver cirrhosis,1 case(8.3%)was peritoneal mesothelioma,1 cases(8.3%)was peritoneal metastatic carcinoma;2 cases appeared abdominal pain after operation,including 1 case of neutrophil ratio increase,symptoms and persistent time of abnormal laboratory indexes did not exceed 24 h,the incidence rate was 8.3%;no complications of abdominal cavity infection,incision bleeding and puncture site fistula occurred.Conclusion The transgastric NOTES for conducting abdominal exploration and peritoneal biopsy in the diagnosis of unexplained ascites has the advantages of small trauma,less complications and rapid postoperative recovery,possesses an important clinical application value.