1.Clinical validation of multiple biomarkers suspension array technology for ovarian cancer
Bingbing ZHAO ; Zhijun YANG ; Qi WANG ; Zhongmian PAN ; Wei ZHANG ; Li LI
Chinese Journal of Obstetrics and Gynecology 2017;52(1):11-19
Objective To investigates the diagnostic value of combined detection serum CCL18, CXCL1 antigen, C1D, TM4SF1, FXR1, TIZ IgG autoantibody by suspension array for ovarian cancer. Methods Suspension array was used to detect CCL18, CXCL1 antigen, C1D, TM4SF1, FXR1, TIZ IgG autoantibody in 120 cases of healthy women, 204 cases of patients with benign pelvic tumors, 119 cases of pelvic malignant tumor patients, and 40 cases with breast cancer, lung cancer oroliver cancer, respectively. Constructed diagnosis model of combined detection six biomarkers for diagnosis of ovarian malignant tumor. Constructed diagnosis model of combined detection autoantibodies to diagnose epithelial ovarian cancer. Analysed the value of detecting six biomarkers for diagnosis of ovarian malignant tumor and detecting autoantibodies for diagnosis of epithelial ovarian cancer. Analysed diagnostic value of detecting six biomarkers to diagnose stageⅠandⅡepithelial ovarian cancer. Compared diagnostic value of detecting six biomarkers in diagnosis of tissue types and pathologic grading with that of CA125. Results Model of combined detecting six biomarkers to diagnose ovarian malignant tumor was logit(P)=-11.151+0.008×C1D+0.011 × TM4SF1+0.011 × TIZ-0.008 × FXR1+0.021 × CCL18+0.200 × CXCL1. Model of combined detection autoantibodies to diagnose epithelial ovarian cancer was logit(P)=-5.137+0.013 × C1D+0.014 × TM4SF1+0.060 × TIZ-0.060 × FXR1. Sensitivity and specificity of detecting six biomarker to diagnose ovarian malignant tumor was 90.6% and 98.7%. Sensitivity and specificity of detecting autoantibodies to diagnose epithelial ovarian cancer was 75.8%and 96.7%. Combined detection for six biomarkers to diagnose serous and mucinous ovarian cancer was statistically no better than those of CA125 (P=0.196 and P=0.602, respectively);there was significantly difference in diagnosis of ovarian cancer (P=0.023), and there was no significantly difference in diagnosis of different pathological grading (P=0.089 and P=0.169, respectively). Conclusions Constructing diagnosis model of combined detection six biomarker to diagnose ovarian malignant tumor and constructed diagnosis model of combined detectionautoantibodies to diagnose epithelial ovarian cancer. Combined detection six biomarkers to diagnose serous and mucinous ovarian tumors is better than that of CA125.
2.The clinical value of serum matrix metalloproteinase-9, heparanase and cathepsin L detection in determining the degree of ovarian cancer invasion and metastasis
Wei ZHANG ; Xingzi YANG ; Qi WANG ; Zhijun YANG ; Hong CHEN ; Sumei WANG ; Zhongmian PAN ; Li LI
Chinese Journal of Laboratory Medicine 2012;35(6):559-563
Objective To explore the clinical value of serum cathepsin L (CL),matrix metalloproteinase-9 (MMP-9) and heparanase (Hpa) detection in determining the degree of ovarian cancer invasion and metastasis.Methods Enzyme-linked inmunosorbent assay (ELISA) and electrochemiluminescencl immunoassay (ECLIA) were used to detect the serum content of MMP-9,Hpa,CL in 217 cases with untreated ovarian cancer before surgery( in FIGO Ⅰ - Ⅱ stage 83 cases,Ⅲ-Ⅳstage 134 cases),100 cases with benign ovarian tumors and 101 healthy women control.All of the patients from Guangxi Medical University Affiliated Tumor Hospital,from September 2003 to October 2009.The relationship between the clinical pathological factors of ovarian cancer and serum content of MMP-9,Hpa,CL was analyzed.On the basis of clinical pathological diagnosis as “gold standard”,the ROC curves was drawed to evaluate the clinical value of serum CL,MMP-9,Hpa combined detection in determining the degree of ovarian cancer invasion and metastasis before surgery.Results The serum content of CL,MMP-9 and Hpat in patients with ovarian cancer were (21.23 ± 8.17),( 193.95 ± 42.49),(7.68 ± 2.32) μg/L respectively,which was higher than that in patients with benign ovarian tumors [ ( 10.97 ± 3.84),( 143.66 ± 28.47),( 4.86 ± 1.37) μg/L respectively ] and normal control [ (5.59 ± 1.75),( 57.99 ± 1 1.42),( 2.77 ± 0.80) μg/L respectively ],there was difference statistically significant ( t value CL was - 13.242,- 13.498 respectively; MMP-9 was - 14.521 and - 21.290 respectively; Hpa was - 10.896 and - 18.280 respectively,P < 0.001).The serum content of CL [ ( 21.59 ± 8.24) μg/L ] in patients with epithelial ovarian cancer ( EOC) was significantly higher than that [ ( 19.57 ± 7.69) μg/L ] in non-epithelial carcinoma ( F =1 1.209,P =0.048).The serum CL,MMP-9 and Hpa content in FIGO Ⅰ -Ⅱ stage patients was (19.66 ± 7.83),(182.63 ±42.30),(7.21 ±2.05) μg/L,which was lower than that (22.64 ±8.31),(202.81 ±39.74),(8.51 ± 1.92) μg/L in FIGO Ⅲ-Ⅳ stage patients ( F value was 12.452,70.565 and 195.122respectively,P value was 0.030,0.002 and 0.000 respectively).In patients with EOC,the serum CL,MMP-9 and Hpa content in eases with poorly differentiated was ( 23.04 ± 7.67),( 200.12 ± 40.82),(8.22 ± 1.92) μg/L respectively,which was also higher than that in cases with high-moderate differentiated [ ( 18.54 ± 7.30),( 173.43 ± 39.37),(7.20 ± 2.51) iμg/L respectively;F value was 24.545,60.286 and 9.077 respectively; P was 0.004,0.035 and 0.001 respectively ].The serum content of CL and MMP-9(22.96 ± 8.41),(200.44 ±43.82) μg/L respectively in patients with invasion and metastasis in the abdominal cavity was higher than that without invasion and metastasis in the abdominal cavity [ ( 19.07 ±7.36),( 181.04 ± 36.10) μg/L,F value was 12.210,18.084 ; P value was 0.030,0.010 ] ; There was statistically significant relatioship between serum levels of Hpa and patients with distant metastasis ( F =9.430,P =0.042).On base of pathological diagnosis as gold standard,ROC curve showed the sensitivity was 60.9% (70/115),69.6% ( 80/115) and 72.2% ( 83/115) and specificity was 57.4% ( 26/62),67.2%(20/62) and 68.9% (19/62),as serum levels of CL,Hpa,MMP-9 preoperative were detected as tumor markers to determine whether there was cancer invasion and metastasis outside the pelvis.Conclusions There is related with CL,MMP-9 and Hpa levels increase and tumor occurrence and progression in ovarian cancer.The serum content of MMP-9,Hpa,CL detection would be certain clinical reference value to determine extent of invasion and metastasis of ovarian cancer before surgery.
3.Clinical evaluation of autoantibody of splice variant of BARD1 in detection of ovarian cancer
Zhijun YANG ; Yanming JIANG ; Guang YANG ; Yuliang RAN ; Zhihua YANG ; Wei ZHANG ; Jieqing ZHANG ; Zhongmian PAN ; Li LI
Chinese Journal of Obstetrics and Gynecology 2008;43(9):680-684
Objective To investigate the value of autoantibody of breast cancer susceptibility 1 associated RING domain (BARD1) splice variant (OV-142) in detection of ovarian cancer.Methods We cloned OV-142 gene into plasmid pET-30b(+).The recombinant protein of OV-142 was expressed in pET30b(+) system and purified. The autoantibody of OV-142 was detected by indirect enzyme-linked immunosorbent assay (ELISA).Results We successfully constructed the recombinant plasmid of OV-142.The recombinant protein was expressed in pET-30b(+) system and purified.The purification rate of the recombinant protein was up to 90%.The relative amount of autoantibody of OV-142 detected by indirect ELISA was analyzed by receiver operating characteristic curve (ROC) and the cutoff value was determined.Combination of the autoantibody IgG of OV-142 and CA125 was analyzed by logistic regression. The sensitivity,specificity and accuracy was 71.4%,89.1%,and 81.9%,respectively,which were higher than IgG (41.3%,84.2%,66.8% ) and CA125( 61.1%,88.0%,77.1% ) when used alone each.Conclusions OV-142 is a splice variant of BARD1.It may be a potential immunotherapy target of ovarian cancer.Detection of autoantibody of OV-142 is a potent complementary tool of CA125 in ovarian cancerdiagnosis.
4.Clinical study of autoantibody spectrum against ovarian cancer associated antigens combined with CA125 in detecting and monitoring ovarian cancer
Zhijun YANG ; Guang YANG ; Yanming JIANG ; Yuliang RAN ; Zhihua YANG ; Wei ZHANG ; Jieqing ZHANG ; Zhongmian PAN ; Li LI
Chinese Journal of Obstetrics and Gynecology 2011;46(2):113-118
Objective To evaluate the clinical value of autoantibody spectrum against ovarian cancer associated antigens combine CA125 in detecting and monitoring ovarian cancer. Methods Circulating IgG, IgM autoantibodies against ovarian cancer associated antigens which included TM4SF1, C1D,TIZ, OV-142,FXR1 and OV-189 were measured by indirect ELISA in serum from 126 patients with ovarian cancer (prior treatment), 42 patients with benign ovarian masses, 142 healthy women. Cut off value of IgG, IgM autoantibodies were determined by receive operating characteristic (ROC) curve. CA125 was measured in serum by immunoradiometric assay (IRMA). We evaluated the clinical value of combining multiple autoantibodies (autoantibody spectrum ), combining autoantibody spectrum with CA125 by binary logistic regresion. The positive ratio of autoantibody spectrum in serum (prior and post treatment ) of 24 synchronization patients with ovarian cancer was analyzed to evaluate the value in monitoring state of illness.Results Our data indicated that serum contains IgG, IgM autoantibodies against ovarian cancer associated antigens. The positive ratio of IgG autoantibodies in serum from ovarian cancer patients and cancer-free patients were 34. 1% - 47. 6% and 13.0% - 19. 0%, respectively ( P < 0. 05 ). The positive ratio of IgM autoantibodies in serum from ovarian cancer patients and cancer-free patients were 39. 7% - 53.2% and 12. 0% -33.2%, respectively (P <0. 05). The positive ratio of IgG autoantibodies against FXR1 and IgM autoantibodies against TIZ,FXR1 and OV-189 in early stage ( Ⅰ - Ⅱ ) ovarian cancer(55.3% ,63.8%,61.7% and 66. 0% ) were significantly higher than those in advanced ( Ⅲ - Ⅳ )ovarian cancer( 34. 2%,39. 2% ,26. 6% ,45.6%; all P < 0. 05 ). Combining five autoantibodies ( TM4SF1 IgG, TM4SF1 IgM, C1D IgG, FXR1 IgG and TIZ IgM ) showed significantly improved sensitivity (75.4%, P < 0. 05 ), lower specificity (78. 3% ,P < 0. 05 ) and similar accuracy (77. 1%, P > 0. 05 ) in detecting ovarian cancer compared to those of CA125 (61.1% ,88.0% ,77. 1% ). But the autoantibody spectrum showed significantly improved sensitivity in classifying early stage (76. 6% ), compared to those of CA125 (51.1% ,P < 0. 05 ).Combining autoantibody spectrum with CA125 showed significantly improved sensitivity ( 85.7% ), specificity (90. 8% )and accuracy (88.7%) in detecting ovarian cancer compared to those of autoantibody spectrum alone ( all P < 0. 05 ), while CA125 ( 61.1%, P < 0. 05; 88. 0%, P > 0. 05; 77. 1%, P < 0. 05 ). The positive ratio of combine the autoantibody spectrum with CA125 was significantly lower in 24 post-treatment serum (42%) compared to the pairing prior treatment serum ( 88%, P < 0. 05 ). Conclusion Combining the autoantibody spectrum against ovarian cancer associated antigens with CA125 can improve sensitivity,specificity and accuracy in detecting early ovarian cancer and may be used to monitoring state of illness.
5.Relationship between hormone therapy in women with ovarian malignancy and prognosis
Li LI ; Zhongmian PAN ; Xinqiu CHEN ; Kun GAO ; Wei ZHANG ; Yuan LUO ; Zhongqiang YAO ; Xinqiang LIANG ; Wei HUANG ; Desheng YAO
Chinese Journal of Obstetrics and Gynecology 2008;43(11):843-848
Objective To explore the relationship between hormone therapy (HT) in women withovarian malignancy and prognosis. Methods HT was used in 31 patients with ovarian cancer after surgery,and 44 eases with ovarian eaneer served as controL The expression of estrogen receptor (ER)α, ERβ andprogesterone receptor (PR) was detected by immunohistoehemieal staining respectively. The level of serumealeitonin and transforming growth factor α (TGFα) was detected by radio-immune and enzyme-linkedimmunosorbent assay pre- or post-surgery, as well as half a year to one year later post-surgery respectively inthese eases. The survival curve of Kaplan-Meier and log-rank test as well as scale risk of Cox model wereused to analyze the relationship between HT and prognosis of ovarian cancer. Results ( 1 ) The results oflog-rank test showed that there was no difference in survival curve of patients with or without HT [ (1108±52), (1086±43) d; P=0.940] ; the results of scale risk of Cox model also showed that HT was not anindependent prognosis factor for patients with HT. (2) There was no relationship with HT and theaccumulated survival in patients with either positive or negative expression of ERa, ERβ and PR in tissue;as well as between HT and the level of serum TGFα pre-, post-surgery, or half a year to one year aftersurgery. (3) The level of serum caleitonin in patients without HT post-surgery half a year to one year laterwas higher than that pre-surgery [ (141±13), (95±11) μg/; P<0.05], but there was no significantdifference between patients with HT half a year to one year later past-surgery and pre-surgery [ (90±18)μg/L, (93±14) μ/L; P>0.05]. (4) There was a significant difference in body and emotion function between HT and without HT groups [(1.84±1.50), (1.45±0.82); (12.69±10.20), (12.90±11.61); P<0.05], as well as in sex quality and autonomic nerve maladjustment and in the special listmade [(1.05±0.74), (1.77±1.08); (10.10±3.21), (13.09±4.30); P<0.05]. ConclusionsThere is no adverse influence on prognosis in using of HT for patients with ovarian cancer after surgery. HTfor patients with ovarian cancer post-surgery can help keep a stable level of scmm calcitonin as well asimprove the quality of life.
6.Effects of blocking DNA-PKcs on autophagy protein expression and proliferation in esophageal squamous cell carcinoma cell line EC109 after irradiation
Jia SUN ; Xiaojing ZHAO ; Ling YUAN ; Yumeng LI ; Nan LI ; Zhongmian ZHANG ; Haoxun WANG ; Na HAN ; Jun MA ; Jian WANG
Chongqing Medicine 2018;47(9):1158-1160,1164
Objective To study the effect of DNA-PKcs blocking on the expression of autophagic proteins,and proliferation of esophageal squamous cell carcinoma cells(EC109)after irradiation(X-Ray).Methods NU7441 was used to inhibit DNA-PKcs and X-Ray radiation treatment were used to treat EC109 cells.Th experiment was divided into 4 groups,including control group, NU7441 group,X-Ray group,X-Ray+ NU7441 group.The expressions of autophagy protein Beclin-1 and LC3B were detected by Western blot.Apoptosis was detected by flow cytometry.MTT assay was used to detect cell proliferation.Results The expression of p-DNA-PKcs in EC109 cells was decreased after NU7441 treatment and increased after X-ray irradiation.Compared with untreat-ed cells(control group),the expressions of both Beclin-1 and LC3B in X-Ray+NU7441 group were increased.Compared with the X-Ray group,the expression of Beclin-1 in the X-Ray+NU7441 group was increased.Compared with the control group,the apoptotic rate of EC109 cells in the X-Ray group and X-Ray+NU7441 group was significantly increased,the difference was statistically significant(P<0.05).Compared with the X-Ray group the apoptosis rate in the X-Ray-NU7441 group was significantly increased.The MTT results showed that compared with the control group,the proliferation in the X-Ray group and X-Ray+NU7441 group was significantly inhibited, the difference was statistically significant(P<0.05).Conclusion NU7441 inhibits the expression of DNA-PKcs protein in EC109 cells, which could promote the expressions of autophagy protein Beclin-1 and LC3B,promotes apoptosis and inhibits cell proliferation.
7.In vitro Mechanical Behaviors Evaluation and Testing Devices of Orthopedic Sutures
Qian ZHANG ; Jiayan LUAN ; Zhongmian XU ; Lu WANG
Journal of Medical Biomechanics 2019;34(5):E560-E566
Orthopedic sutures are vital medical devices in repair surgeries of tendons and ligaments. Properties especially mechanical properties of orthopedic sutures have great impacts on the effects of tendon/ligament repairs. In this paper, clinically used orthopedic sutures including absorbable sutures and non-absorbable sutures were summarized firstly, by exploring literature about orthopedic sutures, and the mostly used orthopedic sutures are non-absorbable sutures. Then, based on the particularity of the repaired sites of orthopedic sutures, the in vitro mechanical test devices and evaluation method of orthopedic sutures were reviewed, mainly including the mechanical properties of sutures, suture-tissue, and suture-anchor. In addition, the characteristics of different evaluation method or test devices were compared. It was still difficult to evaluate the mechanical properties of orthopedic sutures due to the lack of instruments and test criteria.