2.Optimization of a Liquid Chip System for the Detection of Serum Biomarkers of Colorectal Cancer and Its Application
Hua-Ning ZHANG ; Xue-Qin GAO ; Jin-Xiang HAN ; Hai-Nan HUANG ;
China Biotechnology 2006;0(12):-
Liquid chip technology have been licensed to be used in clinic because of its advantage of high-throughput, high-sensitivity, good signal to noise ratio, reaction in liquid phase, convenient operation and short time consuming, etc. The optimization of a liquid chip system for the detection of serum biomarkers of colorectal tumour and initial application in the detection of CEA were studied. The optimized reaction conditions of liquid chip were determined through orthogonal design after it was prepared. The results showed that the consuming reaction time of the coated antibody and the antigen was 1hour. The microspheres, biotinylated detecion antibody and the consuming complexes and avidin-PE time of the microspheres and the biotinylated tested antibody was 1hour, 1hour and 15minutes respectively.the consuming time of the complexes and avidin-PE was fifteen minutes, The optimized dilution of the biotinylated tested detection antibody was 1∶300 and the optimized concentration of avidin-PE was 12?g/ml. Totally 55 clinical samples were detected by the liquid chip and by Enzyme-Linked Immunosorbent Assay (ELISA) simultaneously and the results of the two methods were compared. The results of the two methods showed good correlation between positive and negative samples but the detection limits and the dynamic ranges of the liquid chip method were more sensitive and wider than those of the ELISA. The multiple tumour biomarkers may be detected simultaneously and the time of clinical test and manpower requirements were reduced by the liquid chip method.
3.Direct inhibitory effects of 153Sm-DTPA-c(CGRRAGGSC) on human prostate cancer PC-3 cells
Qing-hua, WU ; Lu, LIU ; Ze-xuan, YANG ; Hai-lin, GAO ; Jin, SUN ; Qi, NIE
Chinese Journal of Nuclear Medicine 2011;31(4):241-244
Objective To investigate the direct inhibitory effects of 153Sm- DTPA-c (Cys-Gly-Arg-Arg-Ala-Gly-Gly-Ser-Cys) NH2 ( 153 Sm-DTPA-c (CGRRAGGSC)) on human prostate cancer PC-3 cells. Methods 153Sm-DTPA-c(CGRRAGGSC) was synthesized by the reaction of 153SmCl3 with DTPA-c(CGRRAGGSC) using indirect synthesis method. PC-3 cells in vitro culture were divided into four study groups, groug A ( the control, with PBS only), group B with 1.5 mg/L c ( CGRRAGGSC), group C with 370 kBq 153 SmCl3 and group D with 370 kBq 153Sm-DTPA-c(CGRRAGGSC). PC-3 cell growth was assayed by 3-(4, 5-dimethylthiazol-2-yl ) -2, 5-diphenyltetrazolium bromide (MTT) method. Cell cycle and apoptosis were analyzed by flow cytometry. The expression changes of interleukin 11 (IL11 ) and IL11 receptor (IL1 1 R) in PC-3 cells were examined by Western Blot. One way analysis of variance (ANOVA) and paired-t test were applied for statistic analysis. Results The labeling yield of 153Sm-DTPA-c (CGRRAGGSC) was 85% and the radiochemical purity was 95.8%. The specific activity of 153Sm-DTPA-c(CGRRAGGSC) was 1.32 × 105 MBq/μmol. Significant inhibitory effects on the growth of PC-3 cells were found in both group C and D, with a time-dependent manner. However, no obvious inhibition was found either in group A or in group B. After 48 h,significant differences of sub-G1 peak area were found among groups, (0. 98 ± 0. 18)%, (0. 35 ±0. 10)%, (4.05 ±0.28)% and (13.38 ±0. 89)% for group A, B, C and D, respectively. Furthermore,sexpression of ILl 1R in group D was significantly lower than that in group B and C with absorbance values 0. 339 ~ 0.014, 0.338 ~ 0.019, 0.226 ~ 0. 015 for group B, C and D, respectively. Absorbance values in groups B and C were not significantly different after treatment, compared with those before treatment; however, there was difference between absorbance values after and before treatment in group D ( t = 0. 405,1. 163,135.989,P>0.05 >0.05, <0.05). Conchluion 153Sm-DTPA-c(CGRRAGGSC) can directly in hibit the cell growth and expression of human prostate cancer cells PC-3.
4.Comparison among different noncontact specular microscopes for the detection of corneal endothelial cell density
Shi-ming, CHENG ; Jin-hai, HUANG ; Yan, LI ; Xin, YANG ; Rong-rong, GAO ; Qin-mei, WANG
Chinese Journal of Experimental Ophthalmology 2012;30(2):150-154
Background Clinical research showed that the corneal endothelial cell density value from different corneal specula microscopies exist diversity.The relevant literature of SP02,Tomey EM-3000 and SP3000P is still seldom up to now. Objective This research was to assess the repeatability of endothelial cell density measurements by SP02,Tomey EM-3000 and SP3000P respectively and the agreement among 3 kinds of endothelial microscopes.MethodsFifty-four healthy volunteers with the age 17-38 years old were included this research.The written informed consent was obtained from each subject before examination.The corneal endothelial cell densities in the right eyes were analyzed with SP02,Tomey EM-3000 and SP3000P respectively for 3 times under the automatic mode,and the analytical procedure of SP3000P measurement were divided into automatic mode SP3000P (A) and manual correction modes SP3000P( M).The repeatability of each specula microscopy was analyzed by calculating the intraclass correlation coefficients (ICC) and coefficient of variation ( CV ),and the 95% confidence intervals and plotting Bland-Altman graphs were used to analyze the agreement among these methods.ResultsThe mean corneal endothelial cell densities in the population <24 years were significantly higher than the ones ≥ 24 years (t =3.692,P<0.05 ),but no statistical difference was found between different gender ( t =0.335,P =0.739 ).The mean corneal endothelial cell densities were ( 3058 ± 260 ),( 2954 ± 229 ),( 2668 ± 258 ),( 2734 ± 268 ) cell/mm2 ; the ICCs were 0.957,0.940,0.972 and 0.972 and the CV were 0.063,0.061,0.056,0.058 for SP02,Tomey EM-3000,SP3000P (A) and SP3000P ( M ) respectively.The 95% confidence intervals were ( - 100.8 - 306.8 ),( 162.6 - 617.4 ),( 109.9-494.1 ) and ( -0.6 - 132.6 ) cell/mm2 for between SP02 and Tomey EM-3000,SP3000P ( A ) and SP02,SP3000P(A) and Tomey EM-3000,SP3000P(A) and SP 3000P(M) respectively.ConclusionsSP02,Tomey EM-3000 and SP3000P(A) have good repeatability in the measurement of corneal endothelial cell density,however the outcome is different.Therefore,it is not interchangeable for the detection of corneal endothelial cell density.The differences of corneal endothelial cell density obtained from these instruments shall be paid high attention for their differences.SP3000P(A) and SP3000P(M) can be used interehangeably and SP3000P(A) is a preferable choice due to its convenience and quickness.
5.Evaluation of binocular minimum thickness of cornea in Chinese myopic population
Xiao-min, WU ; Si-si, CHEN ; Rong-rong, GAO ; Qin-mei, WANG ; Jin-hai, HUANG
Chinese Journal of Experimental Ophthalmology 2012;(11):1018-1021
Background Central corneal thickness (CCT)is an important parameter to evaluate corneal healthy status and is crucial for surgical planning.However,clinical study found that the center of cornea does not correspond to the thinnest point of cornea.Thus,it is essential to characterize the minimum corneal thickness(MCT) and its location.Objective Present study was to determine the thickness and location of MCT and its relationship to the fellow eye using Pentacam High Resolution technique.Methods The 564 eyes from 282 Chinese myopic patients were reviewed in this study.The CCT,MCT,pupillary central corneal thickness(PCT) and x-y coordinate of thinnest point were bilaterally measured.Written informed consent was obtained prior to the ocular biomedical measurement.Results CCT was (540.07±31.78) μm in the right eyes and (539.24±31.06) μm in the lefi eyes; PCT was (540.25±30.75)μm in the right eyes and (539.48±31.00)μm in the left eyes.MCT was (537.87± 31.91)μm in the right eyes and (536.35±31.24)μm in the left eyes,showing significant differences in all the parameters between the right eyes and left eyes expect for PCT(CCT:P=0.046;PCT:P=0.065 ;MCT:P=0.000).The C CT,PCT and M CT were significantly correlated between the right eyes and left eyes (r =0.97,0.97,0.98,P< 0.01).Bland-Altman plot showed a good consistence between the both eyes.The mean distance from the center was (0.50±0.21) mm in the right eyes and (0.56±0.22)mm in the left eyes,showing a significant difference (P =0.000).The difference between CCT and MCT was approximately (2.20±1.74)μm in the right eyes and (2.88±1.75) μm in the left eyes.The location of MCT in both the right eyes and left eyes tended to symmetry along the vertical midline.The distance between the R (x,y) to transformed L (x,y) was (0.29 ± 0.17)mm and the angular distance was (28.28±28.21)degree.Conclusions This study offers a range of MCT and its location in Chinese myopic patients.The difference exists between the CCT and MCT in bilateral eyes.The location of the thinnest point tends to be symmetrical along the vertical midline between the both eyes.The changes of these parameters may be helpful for the diagnosis of some corneal diseases.
7.Clinical significance of serum CEA, CA125 and CA72-4 in gastric carcinoma with peritoneal metastasis
Liqiu SUN ; Hai CUI ; Yan CUI ; Wenbiao JIN ; Yuzhen CUI ; Aihua GAO ; Xionghu SHEN
China Oncology 2017;27(3):191-196
Background and purpose: Peritoneal metastasis of gastric cancer is mainly discovered in the ad-vanced cancer. Nonetheless, the clinical applicability of each tumor biomarker in peritoneal metastasis of gastric cancer is still ambiguous. Therefore, this study investigated the diagnostic value and clinical significance of CEA, CA125 and CA72-4 in gastric carcinoma patients with peritoneal metastases. Methods: A total of 108 gastric carcinoma patients with peritoneal metastases from Jan. 2008 to Dec. 2013 were studied. All patients were diagnosed by imaging, operations and pathological examination, and also received intravenous or intraperitoneal chemotherapy. Serum tumor markers such as CEA, CA125 and CA72-4 were determined during diagnosis and before each chemotherapy. The diagnostic sensitivity of single marker and combined detection with 2 or 3 markers were analyzed. The correlations among the serum tumor markers and clinical pathological factors, chemotherapeutic effects and survival time were analyzed. Results: Positive rates of CEA, CA125 and CA72-4 were 20.4%, 46.3% and 45.4% in gastric cancer patients with peritoneal metastases, respectively. For these patients, the positive rates of CEA/CA125, CEA/CA72-4, CA125/CA72-4 and CEA/CA125/CA72-4 were 54.7%, 52.8%, 69.5% and 79.6%, respectively. The combined detection of 3 tumor markers was much better than single marker detection (P<0.05). Positive rates of CEA, CA125 and CA72-4 were correlated with the ECOG scale (P<0.05). Positive rate of CA125 was associated with ascites (P<0.001), while positive rate of CA72-4 was associated with ovarian metastasis (P<0.05). Median survival time of patients with positive rates of CEA, CA125 and CA72-4 was significantly lower than that of the patients with normal levels of these markers (P<0.05). Compared with pre-treatment, the levels of all three tumor markers significantly declined after three cycles of chemo-therapy (P<0.05). The decline in CA125 level after chemotherapy was significantly correlated with decreased amount of ascites (P<0.05). The tumor markers turned negative after 3 cycles chemotherapy in patients with positive markers upon initial diagnosis, their survival was significantly prolonged (P<0.001). Conclusion: Combined detection of serum CEA, CA125 and CA72-4 can significantly promote diagnostic rate of gastric cancer with peritoneal metastasis, and may be helpful in evaluating chemotherapeutic effects and predicting prognosis.
8.Diagnosis and therapy for benign paroxysmal positional vertigo of the anterior semicircular canal.
Bo GAO ; Hai-tao SONG ; Jin-mei ZHOU ; Xia GONG ; Wei-ning HUANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2007;42(6):428-431
OBJECTIVETo analyse the video-oculographic findings of positional tests and evaluate the efficacy of canalith repositioning procedure (CRP) in patients with paroxysmal positional vertigo ( BPPV) of the anterior semicircular canal (ASC).
METHODSA retrospective study of 31 patients with ASC BPPV. Then the CRP was performed.
RESULTSTwenty-two individuals (70.97%) presented a unilateral positional nystagmus during the Dix-Hallpike test, in 17 individuals had torsional nystagmus component, 5 individuals only had pure positional down beat nystagmus. Nine patients presented bilateral positional nystagmus, 7 individuals had torsional component positional nystagmus, in 2 patients the direction of the torsional component were the same during right and left Dix-Hallpike test, in 4 patients the torsional component were concurrent with positional down beat nystagmus but the direction could not be ascertained clinically, in 2 patients had pure positional down beat nystagmus. Nineteen patients (61.29%) had unilateral lesion, 11 patients had the left ASC BPPV, 8 patients had right ASC BPPV. Eleven patients had with both ASC and PSC BPPV in the ipsilateral. Twenty-one patients (67.74%) were cured, 29 patients (93.55%) were improved, 2 (6.45%) patients were inefficacy. CRP effectively resolved the nystagmus and vertigo in 14 patients (45.16%) when applied only once, The average number of CRP was 1.7 times, there were 5 patients recurrence during the follow-up.
CONCLUSIONSASC BPPV was not a common condition. The torsional nystagmus component of ASC BPPV might be weak during the Dix-Hallpike test. The positional nystagmus of ASC BPPV was triggered bilaterally. Based on these findings, CRP could be one of the most effective treatment methods for ASC BPPV.
Adult ; Aged ; Benign Paroxysmal Positional Vertigo ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Semicircular Canals ; Vertigo ; diagnosis ; therapy
9.Bone marrow mesenchymal stem cells combined with VEGF gene for the treatment of limb ischemia in rabbits
Hai YUAN ; Dianning DONG ; Xing JIN ; Yongxin ZOU ; Xuejun WU ; Xiangqian KONG ; Jingyong ZHANG ; Binbin GAO ; Hua ZHOU
Chinese Journal of General Surgery 2012;27(1):44-47
Objective To evaluate bone marrow mesenchymal stem cells combined with VEGF gene in the treatment of limb ischemia in rabbits.Methods The right hind limb ischemia model of New Zealand rabbit was established by superficial femoral artery excision and deep femoral artery ligation.Rabbits then were divided randomly into 4 groups: empty plasmid control group(EP group),bone marrow mesenchymal stem cells group(BMSC group),VEGF gene therapy group(VEGF group),combination bone mesenchymal stem cells and VEGF gene therapy group(BV group).There were 8 rabbits in each group.Angiogenesis was detected by arteriography on day 28 after treatment and expression of VEGF was detected by immunohistochemical staining on day 30 after treatment.Results There were no differences of collateral vessel count between the EP group,BMSC group and VEGF group.The collateral vessel count in BV group was higher than that of the other three groups.Immunohistochemistry of VEGF showed that the integrated optical density(IOD)in BMSC and VEGF groups increased significantly compared with the EP group; the IOD in BV group was the highest compared with the other three groups.Conclusions Combination bone marrow mesenchymal stem cells and VEGF gene in the treatment of limb ischemia in rabbits can obtain stable and effective expression of VEGF along with significant improvement of limb ischemia.
10.Repeatability and reproducibility of retinal straylight measurement using C-Quant compensation comparison method
Jin-hai, HUANG ; Shi-shi, LIN ; Qin-mei, WANG ; Bo, ZHENG ; Si-si, CHEN ; Rong-rong, GAO
Chinese Journal of Experimental Ophthalmology 2012;30(2):160-163
BackgroundIn an ideal eye there would be no light scattering at all,but the eye media is not optically ideal.Intraocular straylight causes a veil of light and reduction in the contrast of the retinal image and thus decrease the quality of vision.ObjectiveThe present study was to investigate the repeatability and reproducibility of C-Quant straylight meter( Oculus,Germany)in measuring retinal straylight of myopia and post-laser in situ keratomileusis(LASIK) corneas.MethodsThis is a prospective research.The consecutive 35 eyes of 21 myopic patients and 34 eyes of 22 patients who received LASIK were included in this trail.Retinal straylight was measured for 7 times at a period of time and analyzed quantificational to evaluate the repeatability of measurement.Thirty-eight eyes of 19 patients were measured again at 3-7 days for 3 times at a period of time to assess the reproducibility of C-Quant straylight meter.The mean standard difference (SD) and coefficient of variation (CV) were used as the credibility evaluation.This clinical study complied with Helsinki Declaration and the informed consent was obtained prior to the medical procedure.ResultsThe straylight Log(s) of 7 times measurement were all less than 0.95.The mean Log(s) were 0.92±0.12 and 0.93±0.17 respectively in myopia group and post-LASIK group,without statistically significant differences among 7 times measurement( F=0.335,P=0.812;F=1.000,P=0.409).The mean SD for the 7 times measurement was 0.07 Log units.SD and CV increased with the number of measurements.The differences of mean SD and CV between 3 times result and 6 times result were significant different (t =-2.080,P =0.045;t =-2.190,P =0.035 ).No difference was found between different time periods( t =-0.531,P=0.598 ).The difference of the results between two measurements from the same patient was 0.013.ConclusionsC-Quant is a noncontact,noninvade,rapid and convenient method for the measurement of straylight in myopia and post-LASIK eyes due to the high repeatability and reproducibility.