2.Rhesus Model-based Immunological Insights into Endometriosis.
Lili ZONG ; Longxia WANG ; Yali LI
Journal of Medical Research 2006;0(09):-
Objective To Investigate the dynamics of the parameters in the immune system,including T cell,NK cell sub-populations and cytokines,on the experimental endometriosis model in rhesus,in order to study the immunity-related profile in endometriosis.Materials Six female healthy rhesuses with regular menstrual cycle were selected,among them,five for experiment and one as control.During day 8-15 of menstruation,or day 3-5 after the midcycle estrogen peak,the endometrium and the greater omentum were implanted into the extra-uterine pelvic cavity in the experimental group and the control group,respectively.Samples of sera were collected at the time of 8th and 16th day of every menstruation after the implantation.An exploratory laparotomy or laparoscopy was undertaken during the period from day 8 to day 15 of the menstruation in the 2nd month and the 4th month after the implantation for observation of the occurrence of endometriosis.Methods Flow cytometer(FCM),radioimmuno-assay(RIA),electrochemiluminescence(ECL) and enzyme-linked immunosorbent assay(ELISA) were used to detect the dynamics of the expression levels of CD3,CD4,CD8,CD16 and activity of NK cells,the dynamics of the IgG、IgM,of cytokines,including TNF-a、IL-6、IL-8、CA125、HGF and ICAM-1 in peripheral blood of rhesus pre-operation and during the three months of post operation.Results Cellular immune function declined in the rhesus endometriosis model,showing from the changes of the CD3、CD4、NK(CD16) ratios,CD4/CD8 ratio and decrease of the NK cell activity;For the humoral immunity,no significant changes occurred in the levels of IgG、IgM,with little decline.5 During the 4-month period post-operation,the levels of CA125 rised gradually,ICAM-1 and HGF increased significantly,IL-8 increased slightly,TNF and IL-6 increased markedly in rhesuses that formed endometriosis.Conclusion This experiment shows what we observed in human patients for the immunological system changes could also be reproduced in the rhesus model.Therefore such a model can be used in the future to test the interference of the immune system to tackle his medical problem.Finally we found the oncogenis marker CA125 shows a gradual increase during the disease course therefore it can be used for prognosis of the disease.
4.Safety of Ultrasound-guided Multifetal Pregnancy Reduction in the Second Trimester
Chen LI ; Longxia WANG ; Pingping WANG ; Cuixia GUO
Chinese Journal of Medical Imaging 2015;(3):222-225
PurposeTo evaluate the safety and feasibility of transabdominal ultrasound guided multifetal pregnancy reduction performed in the second trimester.Materials and Methods Forty-three women receiving embryo implantation were divided into two groups according to whether they have received multifetal pregnancy reduction including 16 in the reduction group in the second trimester to reduce multiple fetuses to twins, and 27 with twin pregnancy and no reduction. The pregnancy outcome was compared.Results In the reduction group, 1 patient had miscarriage because of intrauterine infection; 1 patient lost one of the retained fetuses in two days after reduction and delivered the remaining fetus. In the other 14 cases, 8 delivered and 6 were still in pregnancy. This group had 17 newborns, all without neonatal death or birth defects. In the non-reduction group, there was 1 spontaneous abortion. In 2 patients, 1 of the twin embryos stopped development. 1 case had a spontaneous abortion, the other 24 cases delivered uneventfully. There were 50 newborns in this group with 3 birth defects and no neonatal death. There was no statistically significant difference in gestational age at delivery, birth weight, and the incidence of maternal and neonate complications (P>0.05).Conclusion Multifetal reduction improves pregnancy outcomes, similar to non-reduction twin pregnancy. It is safe and feasible to perform multifetal pregnancy reduction in the second trimester.
5.Amelioration effect of duodenal-jejunal bypass surgery on insulin-resistance in Goto-Kakizaki rats
Longxia WANG ; Xianzhe ZHAO ; Weiwei QIAO ; Jianzhong GU ; Lixin CHEN
Acta Laboratorium Animalis Scientia Sinica 2015;(2):159-164
Objective To establish a Goto-Kakizaki ( GK) rat model of duodenal-jejunal bypass( DJB) and ob-serve the changes in insulin-resistance after surgery, and to explore the mechanism of DJB surgery in treatment of type 2 di-abetes mellitus.Methods Male Goto-kakizaki diabetic rats(GK,n=36)were used as experiment group and 18 healthy male Wistar rats as blank group.GK rats were randomly divided into two groups: diabetic control group and DJB surgery group ( n=18) .Euglycemic-hyperinsulinemic clamp technique was performed in 6 rats randomly taken from each group at third week, sixth week and ninth week after surgery, respectively.The expression levels of Gck, G6P, PEPCK mRNA in the liver and GLUT4 content on skeletal muscle cell plasma membrance were detected one week after the clamp test. Results In the DJB surgery group at the end of third week and sixth week after surgery, the levels of glucose infusion rates and the expression levels of Gck, G6P, PEPCK mRNA in the liver showed no statistically significant difference as com-pared with the diabetic control group (P>0.05).In the DJB surgery group at the end of 9th week after surgery, the glu-cose infusion rate and expression level of Gck mRNA in the liver were significantly higher, and the expression of G6P and PEPCK mRNA was significantly lower than those in the diabetic control group ( P<0.05 for all) .At the three time points (3rd week, 6th week and 9th week after surgery), there was no significant difference in the GLUT4 content on skeletal muscle cell plasma membrane between the diabetic control group and DJB surgery group ( P>0.05 for all) .Conclusions Our results indicate that the mechanism of DJB surgery improving blood glucose level may be closely related to the amel-ioration of insulin-resistance in the liver, thereby augmenting glucose uptake and inhibiting gluconeogenesis in liver through the regulation of glucose metabolism-related enzymes.There is no significant improvement in insulin-resistance in the skele-tal muscles during the experiment period.This result implies that the effect of DJB surgery on type 2 diabetes is related to the duration of therapy.
6.Ovarian Malignant Teratoma:Ultrasound Findings and Misdiagnosis Analysis
Cuixia GUO ; Longxia WANG ; Wei LIU ; Hong XU
Chinese Journal of Medical Imaging 2013;(10):775-779
Purpose To analyze the ultrasound manifestation of ovarian malignant teratoma, and to evaluate its diagnostic value and explore reasons for its misdiagnosis. Materials and Methods A retrospective study was conducted on the ultrasound findings of 25 patients with ovarian malignant teratoma confirmed surgically and pathologically (22 with immature teratoma, 3 with teratoma with malignant transformation). Their ultrasound imaging features were summarized and the reasons for misdiagnosis were further analyzed. Results The ultrasound imaging of 22 immature teratoma cases were divided into three types, namely, predominant solid components, predominant fluid components and mixed components of fluid and solid. All displayed calcification-like strong echo scattering in the low echo;color Doppler showed rich blood signal, little signal or no blood flow signal. Preoperative ultrasound presented1 case of immature teratoma (4.5%), 8 cases of ovarian tumors (malignant) (36.4%) and 13 cases of other diagnosis (59.1%). As to the 3 cases of teratoma with malignant transformation, the ultrasound imaging showed 2 cases were composed by fluid and solid components and that 1 case was mature cystic teratoma;typical hypoecho suggested malignant transformation. Preoperative ultrasound did not present teratoma with malignant transformation. Conclusion The ultrasound findings of ovarian malignant teratoma seem to be complicated and variable;thus the preoperative diagnosis rate is low though it still shows certain characteristics.
7.Gene mutation in fetuses with short limbs deformity
Yanping LU ; Jing CHENG ; Longxia WANG ; Shujuan WANG ; Lihua XIONG ; Zhiying GAO ; Huijun YUAN ; Yali LI
Chinese Journal of Perinatal Medicine 2012;15(8):490-493
Objective To identify the genetic mechanism of fetuses with short limbs deformity.Methods From Aug.2008 to Aug.2011,ten fetuses with obvious short limbs were found in ultrasound screening performed at 18-24 and (or) 30-32 gestational weeks and underwent artificial induced labor with the patient' consent.Amniotic fluid or cord blood of the fetuses was collected for karyotyping analysis and detection of mutation point of fibroblast growth factor receptor 3 (FGFR3)gene by polymerase chain reaction and gene sequencing.One fetus (case 3) who presented with achondrogenesis underwent sequencing of SLC26A2 and Trip11 gene meanwhile.Results Among the 10 fetuses with short limbs deformity,five cases were found during second trimester and five during third trimester.Nine cases were identified as normal karyotype and one was chimera (46,XY/45,XY,- 18).One fetus carried a rare FGFR3 mutation of c.1108G>T (G370C) and was diagnosed as thanatophoric dysplasia at 21+3 weeks.Three fetus carried c.1138G>A (G380R) mutation and were diagnosed as achondroplasia.These four families had low recurrent risk because no gene mutations were found in the parents.Three mothers of these four fetuses were pregnant again and had normal neonates now.No mutations were found in all gene sequencing in case 3.Conclusions Karyotyping analysis and sequencing of FGFR3 gene could find causative gene mutations and provide genetic counselling and prenatal diagnosis for some fetuses with short limbs deformity.In the third trimester,achondroplasia is the most possible diagnosis when short limbs fetus is found by ultrasound.
8.Value of detection of cell-free fetal DNA in maternal plasma in the prenatal diagnosis of chromosomal abnormalities
Shujuan WANG ; Zhiying GAO ; Yanping LU ; Yali LI ; Yanqin YOU ; Liwen ZHANG ; Longxia WANG ; Hong XU
Chinese Journal of Obstetrics and Gynecology 2012;(11):808-812
Objective To investigate the value of detection of fetal cell-free fetal DNA(cff-DNA)in maternal plasma in the prenatal diagnosis of chromosomal abnormalities.Methods The plasma from 3200 gravidas(singleton with 20.3 ± 3.8 gestational weeks)was collected from April 1st 2011 to May 30th 2012.They were divided into 3 groups:(1)To tally 1720 cases were included in the high-risk serological screening group,in which women were younger than 35 years and got high-risk results in serological screening;(2)To tally 1310 cases were included in the advanced age group,in which women's age was more than 35 years;(3)To tally 170 cases were included in the supplementary group,in which women were younger than 35 years and got low-risk results in serological screening,or women who didn't take serological screening tests.All the 3030 gravidas in group 1 and 2 didn't take invasive prenatal diagnosis because of fear of abortion or short of prenatal diagnosis.Cff-DNA were detected by next generation sequencing in Shenzhen BGI Genomics Center for clinical laboratory.Amniocentesis and karyotype analysis were provided to the positive cases and women with negative results were followed-up by telephone.Results(1)The 3200 cases took cff-DNA detection,and 31 cases got positive results,including 27 cases of trisomy 21 and 4 cases of trisomy 18.Sixteen cases of trisomy 21 and 1 case of trisomy 18 were in the high-risk serological screening group.7 cases of trisomy 21 and 2 cases of trisomy 18 were in the advanced age group.Four cases of trisomy 21 and 1 case of trisomy 18 were in the supplementary group.(2)And the 84%(26/31)cff-DNA detecting positive cases received amniocentesis.In the 27 trisomy 21 positive cases,23 received amnioeentesis and got karyotype of 47XN,+ 21,with the diagnostic accordance rate of 100%.In the 4 cases who didn't take karyotype analysis,fetal anomaly(ventricular septal defect,dextrocardia and choroid plexus cyst)was found in 1 case before 20 gestational weeks;intrauterine fetal demise happened in 1 case before getting the result;2 other cases who already had healthy children took abortion in the local hospital without taking amniocentesis.In the 4 trisomy 18 positive cases,3 took amniocentesis,2 of which were trisomy 18 and took abortion,the other was chimera(46,XN/47,XN,+ 18)with only 2% cells of trisomy 18,with no malformation found after delivery.Hypoevolutism(3 weeks less than gestational week),general hydropsy and intrauterine fetal demise happened before the other case took amniocentesis.(3)Follow up of cff-DNA negative cases:until May 30th 2012,no Down's baby was found in the 1230 cases with cff-DNA test negative results.Conclusions(1)The non-invasive fetal trisomy test(NIFTY)by next generation sequencing is a safe,accurate and high throughput method for the prenatal diagnosis of trisomy-21.(2)Use NIFTY as a further screening for pregnant women with high-risk serological screening results could lower invasive prenatal diagnosis rate.(3)Cases with positive NIFTY test results should receive amniocentesis and karyotype analysis to confirm the diagnosis before abortion.
9.Detection of trisomy 21 by quantitative fluorescent PCR in clinical samples undergoing prenatal diagnosis for hereditary hearing loss
Yanping LU ; Jing CHENG ; Bing HAN ; Longxia WANG ; Pu DAI ; Huijun YUAN ; Yali LI
Chinese Journal of Obstetrics and Gynecology 2011;46(6):427-430
Objective To establish the genetic test technique of trisomy 21 concurrently conducts with prenatal diagnosis for hereditary hearing loss. Methods Fifty-four pregnant women who underwent prenatal diagnosis for hearing loss of their fetuses in Chinese People's Liberation Army General Hospital from March 2009 to May 2010 were enrolled in this study. All probands from the deaf families have confirmed the causative mutation for hearing loss in Genetic Testing Center in Chinese People's Liberation Army General Hospital. The mean age of 54 pregnant women is 31 years at pregnancy of 18 - 26 weeks, 5 cases > pregnancy of 23 weeks, 9 cases ≥ 35 years. All subjects did not conduct the serologic tests for trisomy 21before. Fifteen to twenty ml amniotic fluid was drawn from 49 cases at pregnancy of 18 - 23 weeks and 5 cases > pregnancy of 23 weeks. One to two ml umbilical blood was drawn from 5 cases > pregnancy of 23 weeks. For 9 cases ≥ 35 years, amniotic fluid cell culture and karyotyping analysis were conducted concurrently. A multiple quantitative fluorescent ( QF) PCR and six microsatellite markers were applied to as trisomy 21. Results (1) Fifty-four fetuses were successfully conducted prenatal genetic diagnosis for hearing loss (included GJB2 and SLC26A4). Ten fetuses copied the exactly same genotypes as the probands. The other 44 cases fetuses did not copy the same genotypes as the probands and won't develop hearing loss. The hearing test showed normal hearing for the neonates. (2) All the 54 fetuses were excluded of trisomy 21 by QF-PCR and were verified after birth. Five fetuses with advanced maternal age were performed karyotyping analysis and showed normal. The diagnostic results of QF-PCR can be obtained in 1 - 3 days without misdiagnosed. Conclusions QF-PCR is an efficient, rapid and accurate technique for detection of trisomy 21 without increasing sample amount. It can be used for fetuses who were undertaken hearing loss gene test or other prenatal gene test.
10.Ultrasonography imaging feature of primary fallopian tube carcinoma and the reason of misdiagnosis analysis
Hua WANG ; Longxia WANG ; Qiuyang LI ; Qin LIU ; Yanqiu WANG ; Ping HE
Chinese Journal of Medical Ultrasound (Electronic Edition) 2017;14(2):111-116
Objective To analyze ultrasonographic imaging feature of primary fallopian tube carcinoma (PFTC) and the reasons for misdiagnosis.Methods Clinical data and ultrasonographic imaging feature of 41 patients with pathologically confirmed PFTC were retrospectively analyzed from August 2008 to November 2016 in General Hospital of Chinese People's Liberation Army.Results Ultrasonographic characteristics of 41 PFTC cases:(1) Type Ⅰ (6 cases),the cystic adnexal mass with single or multiple papillary projections and circuity tubular structures,color Doppler flow imaging showed abundant blood flow signal inside the nipples.(2) Type Ⅱ (2 cases),the sausage shaped complex adnexal mass showed clear boundary,the cystic area that lined along the fallopian tube was around or at the side of the solid part,color Doppler flow imaging showed rich or abundant blood flow signal inside the solid part.(3) Type Ⅲ (13 cases),the sausage shaped hypoechoic adnexal mass showed clear boundary,color Doppler flow imaging showed rich or abundant blood flow signal inside the mass.(4) Type Ⅳ (14 cases),the single or multiple adnexal masses showed irregular surface,with predominant solid components,color Doppler showed rich or abundant blood flow signal inside the tumor;the normal ovarian structure was not detected in unilateral or bilateral adnexa area;and one or more signs of metastasis were found,such as the peritoneal thickening of vesicouterine pouch,uterine rectum pouch and omental,metastasis to other distant organs,and so on.(5) Type Ⅳ (6 cases),only hydrosalpinx or no abnormal ultrasonographic changes in the adnexal area.Nineteen (46.3%,19/41) cases were correctly diagnosed by preoperative ultrasonography,while 22 (53.7%,22/41)cases were missed or misdiagnosed.Conclusions Ultrasonography imaging of PFTC has certain characteristics,but it tends to be missed or misdiagnosis when the lesion is small.Ultrasound can show the location,size,internal echo,blood flow and distant metastasis of lesion,which can be taken as the first choice of imaging methods for preoperative diagnosis and postoperative follow-up of PFTC.