Objective To investigate significance and correlation of free fetal DNA (fDNA) and β-human chorionic gonadotropin(β-hCG) in circulation in pregnant women with high-risk of Down's syndrome (DS). Methods Pregnant women with a male fetus at second trimester screening for Down's syndrome were chosen, including 5 women with a trisomy 21 fetus (DS group), 21 women with DS high-risk pregnant women (DS high-risk group) matched with 22 normal pregnant women as control group. Free fDNA in maternal plasma were extracted. Male DYS14 gene was labled as fDNA, real-time PCR was used to detect fDNA expression. The concentration of β-hCG in maternal serum was detected by chemiluminescence immune assay. The relationship between level of free fDNA and β-hCG concentration was analyzed by Pearson correlation analysis. Results (1) The mean level of free fDNA was (127±58 ) GE/ml in DS group, which was significantly higher than (78±28) GE/ml in DS high-risk group and (48±21 ) GE/ml in control group,respectively (P<0.01). When compared the level of free fDNA between DS high-risk group and control group, it reached statistical difference (P<0.01). (2) The mean concentration of β-hCG was (97±43) kU/L in DS group, which was significantly higher than (58±25) kU/L in DS high-risk group and (38±19) kU/L in control group, respectively (P<0.01). The level of β-hCG in DS high-risk group was also significantly higher than control group (P<0.01). (3) The positive relationship between the level of free fDNA in maternal plasma and β-hCG concentration in maternal serum was observed amongthree groups (r=0.83,P<0.05;r=0.76,P<0.01;r=0.86,P<0.01). Conclusions Free fDNA in maternal plasma might be a candidate marker used for prenatal DS screening. However, its clinical value need to be evaluated because of positive correlation between free fDNA and β-HCG in maternal circulation.

Objectives To study the effect and the mechanism of peripheral blood nuclear cells (PBMC) invaded by hepatitis B virus (HBV) on the artificial immunization in newborns Methods Fifty two newborns, whose mothers were hepatitis B surface antigen (HBsAg) positive, were immunized with hepatitis B immunoglobulin and hepatitis B vaccine (HBVac), and then followed for 7 months The newborns′ serum and PBMC HBV DNA was detected by nested PCR, hepatitis B surface antibody (HBsAb) was tested with solid phase radioimmunoassay PBMC from newborn were incubated with PHA and HBsAg The supernatant interleukin 2 (IL 2) level was mesured by enzyme linked immununosorbent assay (ELISA) Results The rate of vaccination failure was higher in the infants with PBMC HBV DNA positive than those with negative ( P

Objective To explore the value of Swansea criteria on diagnosis and severity evaluation of acute fatty liver of pregnancy (AFLP).Methods Fifty-two AFLP patients were admitted to Shandong Provincial Hospital Affiliated to Shandong University between January 1,2000 and December 31,2011.All these cases were retrospectively reassessed by Swansea criteria.According to the severity,prognosis and whether continuous blood purification treatment was needed,these cases were classified as mild and severe cases.Differences between groups were detected by x2 or t test.Bivariate correlation analysis was used for Swansea criteria compliance and postnatal hemorrhage and days in hospital.Results After reassessing by Swansea criteria,31 cases could be diagnosed as AFLP (20 cases met seven or more criteria,11 cases met six criteria)and the other 21 cases could not (16 cases met five criteria,three cases met four,and two cases met three).For the 16 cases that met five Swansea criteria,they were confirmed as AFLP based on postnatal follow-up.The five cases that met four or three criteria were confirmed as AFLP because no other disease could explain their status.Among the patients who met seven or more Swansea criteria,the incidence of intrauterine fetal death was 40％ (8/20),and 65％ (13/20) needed continuous blood purification.These were higher than in patients who met six or fewer Swansea criteria [9％ (3/32) and 28％ (9/32),x2=6.921 and 6.857,P=0.014 and 0.011].Postpartum hemorrhage was positively correlated with Swansea criteria compliance (r2=0.286,P=0.040).Conclusion Patients who meet five Swansea criteria can be diagnosed as AFLP.Swansea criteria can be applied to the severity evaluation of AFLP.

OBJECTIVETo study the effect and mechanism of the peripheral blood mononuclear cell (PBMC) invasion by HBV on artificial immunization in newborns.

METHODSFifty-two newborns of HBsAg positive mothers were immunized with HBIG (hepatitis B immunoglobulin) and HBVac (hepatitis B vaccine) and were followed up for 7 months. The newborns' HBV-DNA in serum and in the PBMCs was detected with nested-PCR; anti-HBs was tested with solid phase radioimmunoassay (SP-RIA). PBMCs isolated from newborn peripheral blood were incubated in the presence of PHA or purified HBsAg. Interleukin-2 (IL-2) level in culture supernatants of activated cells was detected by ELISA.

RESULTSThe failure rate of immunization was higher in infants with positive HBV-DNA in PBMCs than those with negative HBV-DNA (P < 0.05); IL-2 level in PBMC culture supernatants was lower in former than in the latter and in normal controls (P < 0.05). The level of IL-2 in the immunization failure newborns was lower than that in the successfully immunized newborns and in normal controls (P < 0.05).

CONCLUSIONSIntrauterine invasion of PBMCs by HBV is one of the important reasons for immunization failure in newborns. IL-2 production is closely related to the invasion of PBMCs by HBV, which may contribute to the failure of artificial immunization in newborns.

DNA, Viral ; blood ; Hepatitis B Surface Antigens ; immunology ; Hepatitis B Vaccines ; adverse effects ; Hepatitis B virus ; isolation & purification ; physiology ; Humans ; Immunization ; adverse effects ; Immunoglobulins ; immunology ; Infant, Newborn ; Interleukin-2 ; biosynthesis ; blood ; Leukocytes, Mononuclear ; virology

Objective To analyze the difference of laboratory test results between early-onset and late-onset severe preeclampsia and to investigate their clinical application values.Methods Totally 108 blood samples were collected from patients with severe preeclampsia who were diagnosed according to the Diagnostic Standard of Obstetrics and Gynecology(7th Edition) published by People′s Medical Publishing House,in Shandong Provincial Hospital affiliated to Shandong University from March to November 2016,which consisted of 64 early-onset severe preeclampsia before 34 weeks gestation(early onset group) and 44 late-onset severe preeclampsia after 34 weeks gestation(late onset group).In addition,42 women with normal pregnancies as the control group were selected.General clinical data were collected,and the blood sample was analyzed through detecting Hb,PLT,fibrinogen (FIB),D-dimer,AST,ALT,urea,creatinine (Cr),uric acid,CRP,urine protein.The tested results were analyzed and compared.Flow cytometry was used to analyze the proportion of T helper 1 cells(Th1) and T helper 2 cells(Th2),and the ratio of Th1/Th2 was also calculated.All data and F test were performed by use of statistical software SPSS19.0.Results The pre-pregnancy body mass index(29.55±4.49,30.66±5.13,26.62±3.17,F=9.829,P<0.05),diastolic blood pressure[(105.17±14.46)mmHg(1 mmHg=0.133 kPa),(99.80±12.56)mmHg,(74.36±8.42)mmHg,F=82.088,P<0.05],Hb[(123.22±14.38)g/L,(117.03±16.48)g/L,(112.62±11.24)g/L,F=7.133,P<0.05],urea[(6.56±2.36)mmol/L,(4.51±1.35)mmol/L,(3.04±0.87)mmol/L,F=51.733,P<0.05],Cr[(68.47±18.05)μmol/L,(61.37±14.37)μmol/L,(48.54±8.73)μmol/L,F=23.737,P<0.05],CRP[(7.68±8.76)mg/L,(5.88±6.03)mg/L,(3.56±2.41)mg/L,F=4.735,P<0.05],urine protein[(3.66±0.76)g/L,(2.20±1.05)g/L,(0.19±0.40)g/L,F=249.714,P<0.05]had a statistically significant difference among the early-onset,late-onset and control groups.The flow cytometry results demonstrated that the proportion of Th1 in early-onset group(19.83±3.04)was higher than that in both late-onset (14.49±2.79)and control groups(11.78±1.17),on the contrary,the result of Th2 was much lower(early-onset:1.02±0.12,late-onset: 1.11±0.12,control: 1.56±0.11),there was statistical significance among these three groups(Th1: F=135.110,P<0.05;Th2: F=293.687,P<0.05).Conclusions It′s necessary to real-time monitor the laboratory indicators,such as liver and kidney function,especially the immunologic function indicators for evaluating the disease of early-onset and late-onset severe preeclampsia and personal treatment,and for ensuring the health of mother and fetus and improving the prognostic of mother and fetus.

Objective:To investigate the expression of programmed cell death protein 1 (PD-1), programmed cell death ligand 1(PD-L1), and lymphocyte-activation gene 3(LAG-3) in different subsets of lymphocytes and their relationship with the immunologic imbalance in preeclampsia.Methods:We enrolled 25 cases of singleton pregnant women with preeclampsia who were delivered by cesarean section in the Shandong Provincial Hospital Affiliated to Shandong First Medical University from May 2019 to January 2020 as the preeclampsia group. According to the allocation ratio of 1∶1 matched for the date of cesarean section and pregnancy week at delivery, another 25 healthy singleton pregnant women underwent elective cesarean section were selected as the normal group. The decidua tissue was obtained during cesarean section. The expression levels of PD-1, PD-L1, and LAG-3 on decidual T cells, natural killer (NK), and natural killer T (NKT) cells were measured by flow cytometry and compared between the two groups using two independent samples- t test. Results:(1) The expression of PD-1 on decidual T cells and NK cells of the preeclampsia group were lower than those of the normal group (37.84±3.82 vs 57.02±3.89, t=3.529, P<0.001; 3.28±0.48 vs 5.69±0.99, t=2.184, P=0.034), but did not differ significantly in the expression on decidual NKT cells ( P=0.461). PD-L1 expression on decidual NK cells of preeclampsia group was lower than that of the normal group (0.60±0.11 vs 1.32±0.19, t=3.319, P=0.002), but showed no significant difference in the expression level on T cells and NKT cells (both P>0.05). The preeclampsia group was noted for a lower expression of LAG-3 on decidual T cells and NKT cells compared with the normal group (2.32±0.36 vs 4.09±0.67, t=2.335, P=0.024; 35.40±4.97 vs 56.27±4.49, t=3.282, P=0.002), while showed no significant difference in the expression level of NK cells ( P=0.112). Conclusions:The decreased expression of PD-1, PD-L1, and LAG-3 in the decidual lymphocyte subsets may be involved in the immunologic imbalance of preeclampsia through the over-activation of immunocytes at the maternal-fetal interface.

Objective:To evaluate the short-term and long-term efficacy of Heidelberg triangle dissection in surgical treatment for pancreatic head cancer.Methods:The clinicopathological data of 97 patients with pancreatic head cancer who underwent pancreaticoduodenectomy at Cangzhou Central Hospital from Jan 2017 to Jan 2020 were retrospectively analyzed. After propensity score matching, 33 patients were included into the Heidelberg group and 36 patients in the control group.Results:There were no significant difference between Heidelberg group and control group in preoperative general conditions and postoperative major complications, while there were more cases with safe surgical margin > 1 mm (81.8% vs. 58.3%, P=0.034) and a significantly higher total number of lymph node dissection (11.24±2.35 vs. 9.50±2.76, P=0.006). In the survival analysis, the cumulative recurrence rate at 12 months (0.182±0.067 vs. 0.444±0.083, P=0.023) and 18 months (0.424±0.086 vs. 0.667±0.079, P=0.024) were better off in the Heidelberg group, and the local recurrence rate in the Heidelberg group was significantly lower than that in the control group (15.3 % vs. 36.1 %, P=0.037). However, there was no statistical difference in the overall survival time between the two groups. Conclusion:Dissection of Heidelberg triangle in radical resection of pancreatic head cancer reduces tumor local recurrence ,while fails to provide survival benefit.

OBJECTIVETo assess the value of combined fetal karyotyping and chromosomal microarray analysis (CMA) for the verification of high-risk pregnancy signaled by noninvasive prenatal screening (NIPS) based on high-throughput sequencing.

METHODSOne hundred and fifty-one pregnant women with high risks for aneuploidies of chromosomes 13, 18, 21, X and Y or pathological copy number variations (CNVs) by NIPS were subjected to amniocytic karyotyping and CMA analysis.

RESULTSOne hundred and forty-two women were found to have a high risk for fetal chromosomal aneuploidies, which included 83 cases of trisomy 21, 17 cases of trisomy 18, 2 cases of trisomy 13, and 40 cases of sex chromosome aneuploidies. Amniocytic karyotyping and CMA analysis has confirmed 81 cases of trisomy 21, 15 cases of trisomy 18, 10 cases of 47,XXY, 4 cases of 47,XXX, 2 cases of 47,XYY and 1 case of 46,X,del(X)(q26.1). Two trisomy 21, two trisomy 18, 2 trisomy 13, and 23 cases of sex chromosomal aneuploidies were verified as false positives. For 9 women with pathological fetal CNVs detected by NIPS, combined fetal karyotyping and CMA has confirmed 1 case of chromosome 13 microdeletion, 1 case of chromosome 18 microduplication, and 1 case of chromosome 18 deletion. For a case with 30 Mb duplication of chromosome 2 and 25 Mb duplication of chromosome 8, CMA analysis had no positive finding, while fetal umbilical cord blood karyotyping has yielded a 46,XX,dup(2)(p23.1p25.3)[13]/46,XX[87] karyotype. The remaining 5 cases were confirmed as false positive results.

CONCLUSIONCombined fetal karyotyping and CMA has provided a powerful tool for verifying high-risk fetuses signaled by NIPS.

Aneuploidy ; DNA Copy Number Variations ; Down Syndrome ; Female ; High-Throughput Nucleotide Sequencing ; methods ; Humans ; Karyotyping ; Microarray Analysis ; Pregnancy ; Prenatal Diagnosis ; Trisomy 13 Syndrome ; Trisomy 18 Syndrome