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Objective:This study aimed to compare the detection efficacy of transrectal ultrasound-guided transrectal cognitive fusion targeted+ systematic prostate biopsy and transperineal cognitive fusion targeted + systematic biopsy in patients with suspected prostate cancer (PCa). In addition, the relative clinical characteristics of PCa were evaluated.Methods:A total of 385 patients with suspected prostate cancer in the affiliated hospital of Qingdao University from May 2019 to November 2019 were retrospectively analyzed. All patients met the prostate biopsy criterion, who underwent transrectal(n=275)and transperineal(n=110)prostate biopsy respectively. There were no significant differences of mean age [(70.7±7.3)years vs.(69.2±8.4) years], PSA [(55.12±116.96)ng/ml vs. (63.41±315.34)ng/ml], prostate volume [(55.96±35.26)ml vs. (64.35±55.99)ml] between two groups. According to preoperative prostate magnetic resonance imaging combined with intraoperative ultrasound, 2-4 needles targeted puncture of suspected lesion were performed, followed by 12 needle systematic prostate biopsy. The detection rate of prostate cancer between two biopsy ways were compared. The related factors of PCa including age, prostate volume and PSA level were collected for univariable and multivariable logistic analysis. The cancer detection rate was compared and logistic regression was used to assess the impact of patient characteristics on PCa detection.Results:For all patients, the detection rate with cancer between transrectal group and transperineal group were 121/275(40.0%) and 67/110(60.9%), respectively. The transperineal group detected a higher rate of PCa ( P=0.003)and more clinically significant prostate cancers (csPCa) (54.6% vs.36.7%, P=0.001) than that of the transrectal group, there were significant differences between two groups ( P<0.05). Univariate and multivariate logistic regression analysis revealed that PSA( OR=1.025, P=0.001) and prostate volume( OR=0.984, P=0.001)were two independent factors for the detection rate of prostate cancer between two biopsy ways( P<0.05). The effect of age on the detection rate of PCa in the transperieal group was significantly lower than that of the transrectal group( OR=0.037, P=0.238 vs. OR=0.053, P=0.002). Conclusion:The transperieal biopsy could find more PCa than the transrectal biopsy. PSA level and prostate volume could affect the detection rate of cancer between two prostate biopsy ways.
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OBJECTIVE@#To carry out prenatal diagnosis for a fetus with partial 18p deletion detected by non-invasive prenatal testing (NIPT).@*METHODS@#Peripheral blood and amniotic fluid samples of the pregnant woman and her husband were subjected to G-banded chromosomal karyotyping and more accurate chromosomal microarray analysis (CMA). The deletion sites were verified by fluorescence in situ hybridization (FISH) using centromeric probe Cep11 Aqua and telomeric probes Tel11q SO and Tel18 SG.@*RESULTS@#The karyotype of the fetus was determined as 46,XN,del(18)(p11.3). CMA has detected a 6.66 Mb deletion at 18p11.32-p11.31 (136 226-6 796 178). FISH confirmed the presence of a partial deletion at 18p. The mother was found to harbor the same deletion by chromosomal karyotyping as well as CMA analysis. No abnormality was found with the husband.@*CONCLUSION@#Although the fetus and its mother have both carried the same 18p deletion, no clinical manifestation was detected in the mother, which may be attributed to a low penetrance of the disorder. The fetus had died at 33 weeks of gestation with unknown cause.
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Female , Humans , Pregnancy , Chromosome Deletion , Fetus , Genetic Testing , In Situ Hybridization, Fluorescence , Karyotyping , Prenatal DiagnosisABSTRACT
Objective To detect sHLA-G expression in plasma exosomes in patients with colorectal cancer and evaluate its clinical significance.Methods Retrospective study.Plasma was collected from 52 primary CRC patients,20 colorectal polyps patients,20 inflammatory bowel disease patients and 25 healthy donors in the Taizhou Hospital of Zhejiang Province from May 2017 to August 2018.The exosomes were extracted by exoEasyMaxikit and identified by nanoparticle tracking analysis (NTA) and Western blot.Exosomal sHLA-G was detected by flow cytometry (FCM) and enzyme-linked immunosorbent assay (ELISA).The diagnostic values of exosomal sHLA-G detected by FCM and ELISA were assessed,and their diagnostic performances were compared with carcinoembryonic antigen (CEA) and carbohydrate antigen CA19-9 by ROC curve and Youden index.Results The peak size of exosomes extracted from plasma in CRC patients was 101.1 nm and Western blot showed these exosomes expressed marker CD63,CDS1,and TSG101.Exosomal sHLA-G of CRC patients [28.0(21.5-35.1)U/ml] was significantly higher than that in healthy controls[19.6(16.8-21.3) U/ml,U=143.0,P＜0.001],colorectal polyps patients[19.7(16.2-22.5)U/ml,U=180.0,P＜0.001] as well as inflammatory bowel disease patients[19.9(16.7-25.2)U/ml,U=197,P＜0.001].The postoperative sHLA-G level[19.6(17.8-26.3)U / ml,U=325.5,P=0.015] was significantly lower than that in pre-operation.Exosomal sHLA-G was significantly different in different tumor status(U=64.0,P=0.006),lymph node metastasis (U=81.0,P=0.003) and TNM stage (U=105.0,P=0.015) in patients with CRC.ROC curve showed the area under the curve (AUC) of exosomal sHLA-G detected by FCM and ELISA,CEA and CA19-9 was 0.962±0.019,0.899±0.038,0.786±0.058,0.680±0.068,respectively.The difference of AUC was operated by Z test,and it showed that the exosomal sHLA-G detected by FCM was superior to CEA(Z=2.884,P=0.004)and CA19-9(Z=3.994,P＜0.001),and the exosomal sHLA-G detected by ELISA was superior to CA19-9(Z=2.811,P=0.005).Conclusion Plasma exosomal sHLA-G was associated with the progression of CRC and its diagnostic value was superior to the traditional tumor markers CEA and CA 19-9.
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Objective To investigate the roles of human leukocyte antigen-G ( HLA-G) in mye-loid-derived suppressor cell (MDSC) proliferation and M1/M2 macrophage differentiation in C57BL/6-NCI-H446-G tumor-bearing mice for better understanding the mechanisms of HLA-G involved in tumor immune evasion. Methods NCI-H446 ( human small cell lung cancer cells) and NCI-H446-G ( NCI-H446 cells ex-pressing HLA-G) cells were labeled with CFSE at a final concentration of 1μmol/L. CFSE fluorescence lev-els were measured by flow cytometry at different time points. Mouse tumor models were established by subcu-taneous injection of C57BL/6 mice with NCI-H446 and NCI-H446-G cells, respectively. PBS was used to set up negative control group. The mice in each group were sacrificed to collect tissue samples on 5 d, 10 d, 15 d and 20 d after injection. The percentages of splenic CD11b+Gr1+MDSCs, F4/80+CD80+M1 and F4/80+CD206+M2 macrophages were analyzed by flow cytometry. Results Steady expression of HLA-G in NCI-H446-G cells was confirmed by Western blot and flow cytometry. HLA-G enhanced the proliferation of NCI-H446 cells. Tumor size increased dramatically in tumor-bearing mice in the first five days and then de-creased over time. The tumor-bearing mice in the NCI-H446-G group had larger tumor than those in the NCI-H446 group in every time point (P<0. 05) and required longer time to fully reject the tumor. Compared with the PBS and NCI-H446 groups, the percentage of splenic MDSCs in tumor-bearing mice was significantly in-creased in the NCI-H446-G group (P<0. 05). Moreover, the ratio of M1/M2 in NCI-H446-G tumor-bearing mice was much lower than that in the other two groups (P<0. 05). Conclusion This study indicated that HLA-G could increase the percentage of MDSCs and decrease the ratio of M1/M2, which might illustrate the role of HLA-G in tumor immune evasion and its potential clinical significance in cancer immunotherapy.
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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.
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Objective@#To evaluate the clinical value of high-risk humam papillomavirus(HR-HPV) genotyping in diagnosis of cervical lesions.@*Methods@#Between December 2012 and March 2015, a total of 4 095 women who were diagnosed as cervical inflammation-related disease were chosen to be evaluated in gynecological clinic at Taizhou Hospital of Zhejiang Province. All the women experienced HPV genotyping, analysis of the epidemiological characteristics of HPV types in Taizhou area and theresult of histopathologic diagnosis of HPV positive women. Logistic regression analysis was used to estimate the risk of HR-HPV genotyping in cervical lesion progression.@*Results@#Overall, HPV52 was the most prevalent genotype, followed by HPV16, 58, 39 and 56. Among the women with cervical intraepithelial neoplasm (CIN), HPV16 was the most frequent type, followed by HPV52, 58, 33 and 31. Logistic regression analysis showed that a higher risk of CIN2+ for women infected with HPV16 or HPV33, the regression coefficients OR were 3.670(95%CI: 2.399-5.612, P<0.05) and 2.045(95%CI: 1.087-3.848, P<0.05), respectively. Logistic regression analysis showed that a higher risk of CIN2+ for NILM with positive HPV16, and the regression coefficients OR were 2.539(95%CI: 1.622-3.976, P=0.000); the ASCUS with positive HPV16 and 58 had higher risk of CIN2+ , the regression coefficients OR were 1.911(95%CI: 0.530-6.893) and 1.757(95%CI: 0.557-5.548), respectively.@*Conclusions@#HPV genotyping has important clinical value to detect precancerous cervical lesions, especially when cervical cytology is negative, in management of patiemts with atypical squamous cells of an undertermined significance(ASCUS) or low grade squamous intraepithelial lesion(LSIL).
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Objective To investigate the clinical value of serum apolipoprotein A-1 (Apo A-1) detection in HBV-related liver cancer.Methods Totally 362 cases of patients with chronic HBV infection were enrolled from January 2010 to December 2014 in our hospital,including 88 cases of chronic hepatitis B,94 cases of HBV-related liver cirhosis,18 cases of HBV-related liver cancer (without cirrhosis) and 162 cases of liver cirrhosis merged cancer.At the same time,45 cases of healthy people were selected for normal control.The serum Apo A-1,AFP and other laboratory markers were detected,and the test results were statistically analyzed.Results The difference of Serum Apo A-1 and AFP levels in all groups was statistically significant (F =29.86,x2 =112.53,P =0.000).As the disease progressed,the serum levels of Apo A-1 gradually decreased (P ＜ 0.05).But the difference of Apo A-1 level between normal control and HBV-related liver cancer group (without cirrhosis),chronic hepatitis B and liver cirrhosis merged cancer group,liver cirrhosis and liver cirrhosis merged cancer group was not statistically significant (all P ＞ 0.05).The liver cancer patients with Child-Pugh score A,B,C had different serum Apo A-1 levels (all P ＜ 0.05);The serum Apo A-1 level of liver cancer patients with Child-Pugh score A was significantly higher than that of liver cirrhosis (t =-3.02,P =0.003),but the differences of serum Apo A-1 levels between liver cancer and liver cirrhosis patients with Child-Pugh score B and C were not statistically significant (t =0.52,1.19,P =0.610,0.240).The serum Apo A-1 levels of liver cancer patients with TNM stage Ⅰ and Ⅱ were significantly higher than those with TNM stage Ⅲ and Ⅳ (t =3.85,P ＜ 0.001).Conclusion The serum Apo A-1 levels of HBV-related liver cancer patients are related with cirrhosis,Child-Pugh score and TNM stage,and the liver reserve function,the body's stress response and many other factors may contribute to the expression of serum Apo A-1.
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Human leukocyte antigen G ( HLA-G) is a member of the non-classical HLA classⅠb family. It is considered to play a crucial role in immune tolerance. A unique feature of HLA-G is the struc-tural diversity as surface expressed and as secreted molecules, which is mainly attributed to alternative spli-cing of the primary transcript. HLA-G can promote the invasion and metastasis of tumor cells through various ways. In addition, HLA-G has been described included in exosomes. Exosomes released by most cell types are nano-sized vesicular bodies that contain lipid bilayer and rich contents. As a new marker for diseases, exosomes are extensively involved in the occurrence and development of diseases. Recent studies have found that exosomes can express soluble HLA-G, which reveal a new way by which HLA-G regulates tumor micro-environment. In this review, we focus on the expression of HLA-G on exosomes to provide new thoughts for the early detection and treatment of tumors.
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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.
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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.
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Objective To learn the economic operation and the existing problems of county TCM hospitals in Chongqing from 2012 to 2014.Methods 2012-2014 data of annual health financial report were used to conduct descriptive statistics and analysis of the current operation benefit, operating efficiency, debt paying ability, development ability, and cost management ability.Results From 2012 to 2014, the annual incomes of TCM county hospitals in Chongqing were 2.802, 3.412, and 4.385 billion RMB; the annual expenses were 2.813, 3.343, and 4.347 billion RMB; the ratios of income and expense about medical treatment were 0.86, 0.90, and 0.92; the ratios of income and expense of medicine were 1.16, 1.15, and 1.11; the outpatient amounts were 5.45, 5.87, and 6.99 million; the hospital discharge amounts were 330.4, 405.2, and 469.3 thousand; the charges per patient were 153.40, 177.12, and 188.71 RMB; the charges pre bed were 465.83, 533.14, and 571.02 RMB; medical expenses per patient were 4923.26, 5416.77, and 5576.01 RMB; the current ratios were 1.00, 1.06, and 1.09; the asset-liability ratios were 62.25%, 63.79%, and 61.37%; the annual growth rates of total assets were 14.06 % and 24.54%, and the annual growth rates of net assets were 19.44% and 21.70% during 2013 to 2014; 100-yuan hygienic materials and medicine consumption were 51.23, 51.10, and 50.07 RMB during 2012 to 2014; the management fee rates were 18.55%,15.44%, and 14.82%.Conclusion The general economic running of county TCM hospitals in Chongqing is stable and financial balance moves towards rationality; social benefit, expand capacity and cost management ability are enhanced gradually. However, the problems about insufficient government finance input, small medical income elasticity, poor debt paying ability, and diseconomies of scale still exist.
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Objective To study whether there was significant difference between pregnant women , data and the results of prenatal screening of single intrauterine fetal death ( sIUFD) when twin pregnancy and singleton pregnancy for guiding the clinical prenatal screening and risk consulting .Methods By comparative study, 56 cases of sIUFD when twin pregnancy were recorded from 2011 to 2014 in Ningbo Prenatal Diagnosis Center , all were natural pregnancy , the sistens gestational weeks were less than 14 weeks , and 4 993 natural singleton pregnancy .The pregnant women , data and the results of serological prenatal screening between sIUFD and singleton pregnancy were analyzed by t-test and rank sum test .Separately , the 56 cases of prenatal screening , risk value was calculated according to the twins and singleton , then the difference were analyzed combined with the results of follow-up.Results Pregnant women , data of two groups were analyzed, there were no statistically significant difference between sIUFD and singleton pregnancy .The age of sIUFD and singleton was (27 ±3)year-old and (27 ±3)year-old respectively, t=2.56, P>0.05; the weight of sIUFD and singleton was (55.2 ±10.23 ) kg and (56 ±10.34) kg, t=4.268, P>0.05.The gestational weeks of sIUFD and singleton were (39.21 ±0.78)weeks and (39.1 ±0.91) weeks, t=1.3, P>0.05;the weight of newborn was (3.38 ±0.41) kg and (3.31 ±0.43) kg, t=1.9, P>0.05.The AFP multiple of median (AFPMOM) of sIUFD and singleton was 1.41(0.99,1.83) and 1.02(0.84,1.24), Z=5.337, P0.05;unconjugated estriol multiple of median of sIUFD and singleton was 1(0.79,1.16) and 1.01(0.85,1.21), Z=1.334, P>0.05.Trisomy 21 risk of sIUFD and singleton was 7 750(2 200,28 000) and 5 300(2 000,12 000), Z=2.093, P<0.05, that had significant difference.The 56 cases of prenatal screening risk value was calculated according to the twins and singleton , among whom 42 cases had the same conclusion , 14 cases had the different conclusion .Among them, according to singleton calculation , 3 cases for high risk, according to the twin calculation of high risk for 17 cases,χ2 =12.1, P <0.05.According to follow-up, all newborns were normal.Conclusions For the natural pregnancy , sIUFD when twin pregnancy , if the sistens gestational weeks less than 14 weeks, the risk of prenatal screening results calculated according to singleton will be more reasonable , as for the prenatal screening for twin pregnancy , the method needs further exploration .
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Objective To investigate the associations between single nucleotide polymorphisms in the gene encoding human leukocyte antigen-G ( HLA-G) and the genetic susceptibility to high risk human papillomavirus ( HPV) type 18 infection in the subjects from Taizhou, Zhejiang province. Methods The genetic polymorphisms of HLA-G gene (14 bp In/Del and +3142C/G) in cervical samples collected from HPV 18-positive and healthy women were analyzed by PCR and gene sequencing technology. Statistical anal-ysis was performed by using SPSS version 16. 0. Chi-squared test was used to analyze the differences with HLA-G gene allele and genotype frequencies between healthy subjects and patients. Results Compared with healthy subjects, women with oncogenic HPV18 infection showed lower frequencies of -14 bp allele,-14 bp/-14 bp genotype and -14 bp/+3142C haplotype (P<0. 05). Moreover, lower frequencies of+3142C/C genotype were detected in HPV18-infected women with pathologically normal cervix (6. 3% vs 21. 1%, OR=0. 25, P<0. 05) and higher percentages of +3142G/G genotype were detected in women with CIN2/3 stage HPV18 infection as compared with those of the control group (68. 8% vs 35. 1%, OR=4. 06, P<0. 05). Conclusion The HLA-G gene 3′ UTR polymorphisms were closely associated with HPV18 in-fection and cervical intraepithelial neoplasia.
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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 .
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Objective To investigate the relationship between CD158j expression and phosphorylated ERK (p-ERK) in CD4+ CD28null T cells in cerebral infarction (CI) patients- with carotid atherosclerosis and its effects on carotid atherosclerotic plaque stability. Methods Percentage of peripheral CD4+ CD28null and the expression of CD158j and perform on CD4+ CD28null cells was analyzed with flow cytometry in 106 CI patients with carotid atherosclerosis, 33 CI patients with normal carotid arteries and in 50 normal controls, respectively; p-ERK expression was assayed with flow cytometry in 36 CI patients with unstable plaque, and serum IFN-γ was detected with ELISA. The intima-media thickness (IMT) of bilateral carotid arteries in all subjects was confirmed by the colour Doppler ultrasonograph imagingResults Percentage of the CD4+ CD28null T cells, expression of CD158j and perform on CD4+ CD28null T cells and the serum IFN-γ levels was dramatically higher in CI patients than that in normal controls, respectively (all P <0.01), which was decreased in an order of CI patients with patients with unstable plaque, stable plaque, carotid artery IMT and with normal carotid artery. A strong positive correlation was observed between the CD158j expression and degree of p-ERK in CI patients with unstable plaque (P < 0. 01). Conclusion CD4+ CD28null T cells were significantly increased in CI patients with carotid atherosclerosis. CD158j might up-regulate p-ERK expression and induce the proliferation of the CD4+ CD28nullT cells; consequently, higher cytokine production such as IFN-γ produced by CD4+ CD28null T cells may cause the formation of unstable plaque.
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Objective To investigate the characteristics of immunophenotyping and clinical significance of MRD analysis in MM patients. Methods Multi-parameter flow cytometry was applied to analyze the immunophenotyping of malignant plasma cells from 172 MM patients, and normal plasma cells from 16 healthy individuals. MRD was analyzed in 32 MM patients with remission. Meanwhile, the effects of MRD status on the disease relapse and patient disease free survival ( DFS ) time was evaluated by following up patients. Results The immunophenotyping of normal plasma cells were CD38, CD138, CD19 and CD45 positive, while the predominant phenotype of MM cells were CD+38( 100.0% ), CD+138( 100.0% ), CD-19 ( 167/172, 97. 1% ), CD+56( 152/172, 88.4% ) and CD-45( 166/172, 96. 5% ). The characteristic markers for MM cells were CD+38, CD-138, CD-19, CD-45 and CD+56. MRD analysis revealed that, among 32 MM patients with remission, 14 patients were MRD negative and 18 patients were MRD positive. During follow-up of 2 to 16 months, the relapse rate in MRD negative patients was significantly lower ( 4/14, 28.6% ) than that of MRD positive patients ( 13/18, 72. 2% ;χ2 =6. 03, P ＜0. 05 ). Furthermore, the DFS time was significantly longer in MRD negative patients[ 16. 23( 10. 37-21.62 )months ] than that of the MRD positive patients [ 10. 07( 3. 79-16. 20 )months,χ2 =7. 53,P ＜0. 05 ]. Conclusions CD+38, CD+138, CD-19, CD-45 and CD+56 are the characteristic markers of MM cells compared to those of the normal plasma cells. MRD analysis is a valuable prognostic factor for MM patients.
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Objective To explore issues of the human tissue biobank, ranging from its establishment, collection and preservation of samples, quality control, management and application. Methods Development of standardized operational procedures, collection of such samples as fresh frozen tissues from the surgery, paraffin-embedded tissues, whole blood, serum, plasma, and cerebrospinal fluid. Specimens were classified and made aliquots according to different requirements, and then stored at room temperature, -80℃ refrigerator or in liquid nitrogen. Microsoft Access database system was used in the management of these specimens. Results From September 2004 to September 2008, a total of 11 872 samples from patients with benign and malignant diseases were collected and preserved. Among them, 4360 tubes of fresh frozen tissue samples from 2500 cases were provided within and beyond the province. These samples were also applied to DNA, RNA, protein extraction and tissue microarray, and immunohistochemistry-related research. Conclusion Human tissue biobank is highly useful in sharing human tissue resources effectively, as it can provide high quality specimens from benign and malignant diseases and normal control. In addition, it plays a very important role in exploring pathogenesis, developing new technologies for early detection of disease and new therapeutic strategies.
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Objective To establish the expression of membrane-bound HLA-G1-G4 isoforms in choriocarcinoma cell line JAR and to investigate its roles in NK cytotoxicity in vitro. Methods Stable expression of HLA-G1, -G2, -G3 and -G4 in JAR cells was established by gene cloning and transfection.HLA-Gtranscripts and protein expression in the transfected JAR cells was tested by RT-PCR, flow cytometry, Western blot and immunocytochemistry, respectively. High-affinity peptide KIPAQFYIL pulsing was performed to evaluate its effects on HLA-G expression. Effects of HLA-G1-G4 isoforms on NK cytotoxicity was performed with lactic dehydrogenase (LDH) releasing method. Results RT-PCR, Western blot and immunocytochemistry results showed that exogenous HLA-G1-G4 gene were successfully transfected and proteins were stably expressed in the HLA-G negative JAR cells; Flow cytometry data showed that only HLAG1, but not HLA-G2-G4 isoform was detectable in those transfected JAR cells and the peptide pulsing did not affect their expression status. However, all HLA-G1-G4 isoform expressed JAR cells could significantly decreased the NK cell cytotoxicity (P＜0.05). Conclusion HLA-G1-G4 isoform expression could dramatically inhibit NK-92 cell lysis, indicating that membrane-bound HLA-G isoforms are importantly immunotolerant and may play immune regulation roles in various physio-pathological situations.
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Objective To study the effects of cryopreservation on the immunogenicity of human umbilical vein endothelial cells(HUVEC).Methods HUVEC were isolated ex vivo and cryopreserved.Lymphocyte stimulation index(SI)was analyzed by MTT in lymphoeyte-endothelial cell co-culture.Both HLA-ABC and HLA-DR antigen expression on fresh or cryopreserved HUVEC,and the effects of IFN-γ treatment on HLA antigen expression in both groups were determined by using flow cytometry.Results No difference in SI was observed between fresh prepared and cryopreserved HUVEC(1.716±0.181 vs 1.686±0.145,P>0.05).The percentage of HLA-ABC expression was(96.6±1.9)%and(96.0±1.4)%in fresh and cryopreserved HUVEC(P>0.05),and the mean intensity for HLA-ABC expression was 84.1±5.7 and 82.4±4.8 in fresh and cryopreserved HUVEC(P>0.05),respectively.However,no HLA-DR expression was observed in both groups.When treated with IFN-γ,HLA-ABC expression was significantly up-regulated,and HLA-DR expression was induced in a dose-dependent manner.No significant difference was found in the HLA-ABC expression between fresh and cryopreserved HUVEC(P>0.05),while the HLA-DR expression in cryopreserved HUVEC was remarkably lower than in fresh HUVEC with the increase of IFN-γ(P<0.01).Conclusion The immunogenicity of HUVEC remains stable by cryopreservation without IFN-γtreatment or treated with low concentration of IFN-γ(≤50 U/ml).However,the HLA-DR expression in HUVEC was remarkably reduced in eryopreserved cells treated with a high concentration of IFN-γ(≥100 U/ml).These data indieated that the effects of cryopreservation on immunogenicity of HUVEC may result from the decreased responses of HLA-DR expression by the stimulation of IFN-γ treatment.
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Objective To investigate the mechanism of different HLA-G isoform mRNA patterns in different cells alters its cell membrane expression.Methods RT-PCR was used to analyze HLA-G isoform mRNA(HLA-Gl-6)of ovarian cancer cell lines HO-8910,HO-8910PM and OVCAR-3,leukemia cell lines Jurkat,K562,HIJ60,MUTIZ-1,and the chorioeareinoma cell lines JEG-3,JAR.HLA-G between cellular membrane and intracellular expression were analyzed by flow cytometry.Results All HLA-G mRNA isoforms were observed in the positive control cell line JEG-3,but none in the negative control cell line JAR.HLA-G1 isoform mRNA was expressed in HO-8910,HO-8910PM,OVCAR-3,MUTZ-1 and Jurkat cells.HLA-G2 mRNA was not detected in any cell line but JEG-3.HLA-G3 mRNA was found in HO-8910,HO-8910PM,K562,HIJ60,MUTZ-1,OVCAR-3 and Jurkat cells.HO-8910,HO-8910PM,HIJ60,Jurkat cells expre8sed HLA-G4 mRNA.Only the Jurkat cells expressed HLA-G5 mRNA.FACS results showed that JEG3 and HO-8910PM cells membrane expressed HLA-G,however,the intraeellular HLA-G expression was detected in all tested cells except the negative control cell JAR.Conclusion Only the HLA-G1 isoform could be exDressed on cell membrane in particular cell lines. Other isoforms including HLA-C2,-G3,-G4,-G5 and HLA-G6 could not reach cell snrface.