Objective To develop an efficient method for detecting the short tandem repeat(STR) of fetal DNA in maternal plasma by miniSTR technique.Methods A total of 9 blood samples form pregnant women from 11 to 27 weeks of gestation were collected.Each isolated total plasma DNA was amplified in single multiplex using the ABI MiniFilerTM kit,which could simultaneously genotype the 9 miniSTR loci,including D13S317,D7S820,D2S1338,D21S11,D16S539,D18S51,CSFIPO,FGA and Amelogenin,and the PCR products were detected by using ABI PrismTM 3100 DNA Sequencer.The allelic designation of each STR locus was accomplished using the GeneMapper ID 3.2 software.Results Father-origin fetal STR allele was detected in all the 9 plasma DNA samples.An average of 3.1 fetal STR alleles of the 8 autosomal STR loci was observed in each of the 9 plasma DNA samples.As for the Amelogenin locus,Amelogenin Y allele was detected in 5 plasma DNA samples from pregnancies with male fetus,and allelic peak height values were all over 50 RFU,according to ABI Mini FilerTM PCR conditions,and the ratio of Amelogenin Y allele peak height value to Amelogenin X allele peak height value was 8.45%.However,no Amelogenin Y allele was detected in other 4 plasma DNA samples from pregnant women with female fetus.Conclusion The miniSTR technique is suitable for STR genotyping using fetal DNA in maternal plasma,and it suggests a broad application in noninvasive molecular prenatal diagnosis.

Exosomes are vesicle-like structures generated and secreted actively by all kinds of cells.They contain proteins, nucleic acids and also lipids from the original cells, functioning in interactions between cells by shuttling cellular cargoes, and regulation of immune response in the microenvironment.Besides, they play important roles in the oncogenesis and progression of lung neoplasms.Considerable studies have demonstrated great potentials of exosomal proteins and microRNAs as biomarkers for diagnosis, prognosis prediction and treatment of lung cancers, including immunotherapy and targeted delivering of anti-tumor drugs based on exosomes.

Objective To study the molecular genetic background of Bel subtype at ABO blood group.Methods Three samples and fifteen samples were diagnosed as Bel subgroup and normal control samples by serological test,respectively.The extracted DNA was genotyped by sequence specific primer- polymerase chain reaction foilowed by sequencing for Exon6 and exon7 at ABO locus and clones were sequenced.Results A novel Bel variant allele(GenBank EF117687) was identified in a Bel individual.The Bel allele was different from the regular B101 allele by single 952G>A missense mutation in exon7.resulting in an amino acid subsfitution of Val for Met at 318 locus.No mutations were detected in the fifteen control samples and the other two Bel allele samples.Conclusions The mutation position was fimt found to lie on coding region of ABO gene behind nucleotide 930.The mutation of G952A in the al,3 galactosyhransferase gene may be one of the molecular genetic basis of Bel ohenotype.

Objective To investigate serological blood typing of the ABO locus which contradict to general law of inheritance in parentage,and the underlying reasons for severe hemolytic disease of newborn(HDN).Methods To research the family whose newborn is AB phenotypes,mother is O phenotypes and father is AB phenotypes.The familiy were genotyped by parentage tests, serological tests,PCR-SSP and direct DNA sequencing at exons 6 and 7 of ABO gene.At the salne time,HDN was detected by micro column gel Coombs (MGCT), and the primary fingerposts of the routine blood tests. Biochemical tests were dynamically observed.Results The results of parentage tests showed that three-generation pedigree have parent-child relationship. The red blood cell(RBC)of this AB phenotypes of this family members strongly agglutinated(4+)with diverse monoelonal anti-A and anti-B antibodies,and their serum did not contain anti-A and anti-B antibodies in blood anti-typing.PCR-SSP can not detect their A and B gene,but DNA sequencing at exons6 and 7 of ABO gene revealed that it had the B(A)803C→G mutation.Conclusions The genetm basis of this parentage are B(A)803G blood gene which harbored both A and B difunctionality of glyeosyhransferases.This was the first report that severe HDN resulting from a large number of A and B antigens in RBC of B(A)phenotype of a newborn,which has clinical significance on ABO locus.

Objective:To investigate the value of serum microRNA-92a (miR-92a) and microRNA-146a (miR-146a) expression levels combined with lung ultrasound score (LUS) in predicting the severity and prognosis of acute respiratory distress syndrome (ARDS).Methods:116 patients with ARDS admitted to Danzhou People's Hospital from January 2017 to March 2020 were enrolled. On the day of admission, the expression levels of serum miR-92a and miR-146a were detected by real-time fluorescent quantitative reverse transcript-polymerase chain reaction (RT-PCR), and pulmonary ultrasound examination was performed in 12 lung regions, with the total score as LUS score. The difference of each index was analyzed among the ARDS patients with different 28-day prognosis (survival group and death group) and different severity [mild group: 200 mmHg < oxygenation index (OI) ≤ 300 mmHg (1 mmHg = 0.133 kPa), moderate group: 100 mmHg < OI≤200 mmHg, severe group: OI≤100 mmHg]. Multivariate Logistic regression was used to analyze the risk factors of death in patients with ARDS. Receiver operating characteristic (ROC) curve was drawn to analyze the value of miR-92a and miR-146a combined with LUS score in predicting the death of patients with ARDS.Results:116 ARDS patients were included, 39 cases in the death group, 77 cases in the survival group; 20 cases in the mild group, 38 cases in the moderate group and 58 cases in the severe group. The expression levels of serum miR-92a, miR-146a and LUS score in the death group were significantly higher than those in the survival group [miR-92a (2 -ΔΔCt): 3.75±1.64 vs. 2.10±0.78, miR-146a (2 -ΔΔCt): 1.93±0.72 vs. 0.76±0.20, LUS score: 25.80±4.75 vs. 13.40±3.60, all P < 0.01]. With the aggravation of ARDS patients, the expression levels of serum miR-92a and miR-146a and LUS score gradually increased ( F values were 8.115, 6.740 and 6.216 respectively, all P < 0.01). The expression levels of serum miR-92a, miR-146a and LUS score in severe group were significantly higher than those in the moderate group and mild group [miR-92a (2 -ΔΔCt): 3.65±1.62 vs. 2.87±1.16, 1.94±0.68; miR-146a (2 -ΔΔCt): 1.85±0.58 vs. 1.30±0.51, 0.68±0.17; LUS score: 24.15±4.65 vs. 18.60±4.20, 12.20±3.15, all P < 0.01]. Multivariate Logistic regression analysis showed that low OI [odds ratio ( OR) = 2.748, 95% confidence interval (95% CI) was 1.913-6.225, P = 0.024], high LUS score ( OR = 1.685, 95% CI was 1.183-2.758, P = 0.016), high expression levels of serum miR-92a ( OR = 2.560, 95% CI was 1.806-5.627, P < 0.001) and miR-146a ( OR = 1.984, 95% CI was 1.375-3.816, P = 0.008) were independent risk factors for the death of ARDS patients. ROC curve analysis showed that the area under ROC curve (AUC) of patients with ARDS predicted by miR-92a and miR-146a combined with LUS score was significantly higher than that predicted by the three alone (0.918 vs. 0.842, 0.825, 0.807, all P < 0.01), and the sensitivity (94.0%) and specificity (85.2%) were higher. Conclusion:The expression levels of serum miR-92a, miR-146a and LUS score are related to the severity and prognosis of the patients with ARDS, and the combination of the three indicators has better value in predicting the prognosis of the patients with ARDS.

Objective:To assess the diagnostic performance of serum anti-toxocara immunoglobulin G (anti-T-IgG) in ocular toxocariasis (OT) patients.Methods:A diagnostic tests. A total of 109 patients (109 eyes) with clinically-suspected OT who treated in Department of Ophthalmology of Xuzhou First People’s Hospital from June 2015 to December 2022 were included. Patients were divided into two groups, 76 with OT and 33 with non-OT, according to the clinical manifestations and Goldmann-Witmer coefficient. Paired serum and intraocular fluid samples from each patient were collected and analyzed for specific anti-T-IgG using enzyme linked immunosorbent assay. Mann-Whitney test was performed for comparison between groups. The area under the receiver operating characteristic curve (ROC) was used to assess the diagnostic performance of serum anti-T-IgG. Kappa analysis was performed to examine the consistency of serum or intraocular fluid anti-T-IgG positive rate with OT diagnostic result. Spearman’s rank correlation test was performed to assess the association.Results:Compared with the non-OT group, the proportions of children and history of exposure to cats and dogs ( χ2=9.785, 12.026) were significantly higher in OT group, and the differences were statistically significant ( P<0.01). The positive rate ( χ2=24.551) and U value ( Z=-4.379) of serum anti-T-IgG in OT group were higher than those in non-OT group, and the differences were statistically significant ( P<0.000 1). The recommended serum anti-T-IgG cut-off value of 11 U had 0.72 sensitivity, 0.79 specificity, 0.89 positive predictive value, 0.55 negative predictive value, and 0.77 area under the ROC with 95% confidence interval ( CI) 0.669-0.860. Correlation analysis showed that serum anti-T-IgG was positively correlated with intraocular fluid anti-T-IgG ( r s=0.520, 95% CI 0.363-0.648, P<0.000 1). The Kappa values of serum and intraocular fluid anti-T-IgG positive rate with OT diagnosis were 0.457 (95% CI 0.292-0.622) and 0.711 (95% CI 0.582-0.840), respectively. The Kappa value of serum anti-T-IgG positive rate with OT diagnosis was lower than that of intraocular fluid. Conclusion:The sensitivity and specificity of serum anti-T-IgG and the consistency between serum anti-T-IgG positive rate and OT diagnosis are low, suggesting that serum anti-T-IgG level cannot be used as a basis for OT diagnosis.

Objective    To explore the efficacy of a novel detection technique of circulating tumor cells (CTCs) to identify benign and malignant lung nodules. Methods     Nanomagnetic CTC detection based on polypeptide with epithelial cell adhesion molecule (EpCAM)-specific recognition was performed on enrolled patients with pulmonary nodules. There were 73 patients including 48 patients with malignant lesions as a malignant group and 25 patients with benign lesion as a benign group. There were 13 males and 35 females at age of 57.0±11.9 years in the malignant group and 11 males and 14 females at age of 53.1±13.2 years in the benign group. e calculated the differential diagnostic efficacy of CTC count, and conducted subgroup analysis according to the consolidation-tumor ratio, while compared with PET/CT on the efficacy. Results     CTC count of the malignant group was significantly higher than that of the benign group  (0.50/ml vs. 0.00/ml, P<0.05). Subgroup analysis according to consolidation tumor ratio (CTR) revealed that the difference was statistically significant in pure ground glass (pGGO) nodules 1.00/ml vs. 0.00/ml, P<0.05), but not in part-solid or pure solid nodules. For pGGO nodules, the area under the receiver operating characteristic (ROC) curve of CTC count was 0.833, which was significantly higher than that of maximum of standardized uptake value (SUVmax) (P<0.001). Its sensitivity and specificity was 80.0% and 83.3%, respectively. Conclusion     The peptide-based nanomagnetic CTC detection system can differentiate malignant tumor and benign lesions in pulmonary nodules presented as pGGO. It is of great clinical potential as a noninvasive, nonradiating method to identify malignancies in pulmonary nodules.

Objective    To explore the diagnostic value of circulating tumor cells (CTC) measured by magnetic nanoparticle method in lung cancer. Methods    (1) We measured binding capability of A549 or NCI-H1965 cell lines with recognition peptide and capture efficiency by adding tumor cells into the whole blood of healthy human. (2) We measured CTC of 34 patients suspected with lung cancer, and the counting results of CTC were compared with the following pathological results. Results    (1) The binding capability was 80.0%±6.0% for A549 and 70.1%±4.8% for H1957, while the capture efficiency was 57.3%±7.0% for A549 and 37.3%±6.1% for H1975. (2) CTCs were identified in 71.9% of patients with lung cancer. The specificity was 83.3%, and area under receiver operating characteristic (ROC) curve was 0.792 (P=0.003). Conclusion    CTC measured by magnetic nanoparticle method has promising application in the diagnosis of lung cancer.

Lung cancer is the most frequent cancer and the leading cause of cancer death all around the world. Anti-programmed cell death 1 (PD-1)/programmed cell death-ligand 1 (PD-L1) therapies have significantly improved the outcomes of non-small cell lung cancer (NSCLC) patients in recent years. However, the objective response rate in non-screened patients is only about 20%. It is very important to screen out the potential patients suitable for immunotherapy. Immunohistochemical staining of tumor tissue biopsies with PD-L1 antibodies can predict the therapeutic response to immunotherapy to some extent, but it still has some limitations. Recently some clinical studies have shown that PD-L1 expression in circulating tumor cells (CTC-PD-L1) is a potential independent biomarker and may provide important information for immunotherapy in NSCLC. This article will review technology for CTC-PD-L1 detection and the predictive value of CTC-PD-L1 for immunotherapy in NSCLC and review the latest clinical research progress.