Objective To elucidate the prediction ability of monocyte monolayer assay(MMA)used in hemolytic dis-ease of fetus and newborn(HDFN)caused by IgG anti-M.Methods Plasma from eight pregnant women containing IgG an-ti-M were collected,and were divided into two groups(4 cases with HDFN,with severe clinical symptoms such as fetal hy-drops,and 4 cases without HDFN)according to the clinical outcomes.M antigen positive cells were sensitized with dithioth-reitol(DTT)treated plasma from eight pregnant women respectively.MMA was performed by coincubation with monocytes and sensitized M cells,along with negative and positive control set up.T-test was conducted to compare the difference in phagocytic efficiency between two groups.Results The phagocytic efficiency in group with HDFN were 15.37%,13.05%,9.17%and 24.50%respectively,with the mean value of 15.52%,while the group without HDFN were 8.74%,11.07%,5.12%and 6.23%respectively,with the mean value of 7.79%.There was no significant difference in phagocytic efficiency between two groups(P>0.05).The mean values of both groups were not significantly different from the negative control(P>0.05),but both were significantly lower than positive control(P<0.05).Conclusion The low phagocytic efficiency couldn't convince that the MMA is an effective predictor for the HDFN caused by IgG anti-M,indicating that another mech-anism might be responsible for it rather than monocyte phagocytosis.The assessment of the peak systolic velocity in middle cerebral artery of the fetal should be considered in the management for pregnant women who produce IgG anti-M to estimate the situation of fetal anemia.

Objectives:To investigate the clinical and genetic features of young Chinese patients with myeloproliferative neoplasms (MPN) .Methods:In this cross-sectional study, anonymous questionnaires were distributed to patients with MPN patients nationwide. The respondents were divided into 3 groups based on their age at diagnosis: young (≤40 years) , middle-aged (41-60 years) , and elderly (>60 years) . We compared the clinical and genetic characteristics of three groups of MPN patients.Results:1727 assessable questionnaires were collected. There were 453 (26.2%) young respondents with MPNs, including 274 with essential thrombocythemia (ET) , 80 with polycythemia vera (PV) , and 99 with myelofibrosis. Among the young group, 178 (39.3%) were male, and the median age was 31 (18-40) years. In comparison to middle-aged and elderly respondents, young respondents with MPN were more likely to present with a higher proportion of unmarried status (all P<0.001) , a higher education level (all P<0.001) , less comorbidity (ies) , fewer medications (all P<0.001) , and low-risk stratification (all P<0.001) . Younger respondents experienced headache (ET, P<0.001; PV, P=0.007; MF, P=0.001) at diagnosis, had splenomegaly at diagnosis (PV, P<0.001) , and survey (ET, P=0.052; PV, P=0.063) . Younger respondents had fewer thrombotic events at diagnosis (ET, P<0.001; PV, P=0.011) and during the survey (ET, P<0.001; PV, P=0.003) . JAK2 mutations were found in fewer young people (ET, P<0.001; PV, P<0.001; MF, P=0.013) ; however, CALR mutations were found in more young people (ET, P<0.001; MF, P=0.015) . Furthermore, mutations in non-driver genes (ET, P=0.042; PV, P=0.043; MF, P=0.004) and high-molecular risk mutations (ET, P=0.024; PV, P=0.023; MF, P=0.001) were found in fewer young respondents. Conclusion:Compared with middle-aged and elderly patients, young patients with MPN had unique clinical and genetic characteristics.

【Objective】 To identify the antibody specificity in a pregnant women who had no history of blood transfusion but presented the antibodies against high-frequency antigens. 【Methods】 ABO, RhD blood group antigens were identified by saline. Antibody screening and identification were performed by saline and indirect Coomb’s technique. Further antibody identification tests were conducted using papain, trypsin and chymotrypsin-treated cells. Antibody titer in serum was tested. PCR amplification and sequencing analysis of 16 exons of ABCG2 gene were conducted. 【Results】 The blood type of the patient were B, RhD positive. The serum reacted with antibody screening/identified cells by indirect antiglobin test(both 2+ ) but not by saline. The agglutination was enhanced after papain treatment (4+ ), but remained unchanged after trypsin and chymotrypsin treatment (2+ ). The IgG titer was 1∶2. The sequencing analysis of ABCG2 gene revealed a homozygous nonsense mutation(c.376C>T, p. Gln126X) in exon 4 of the women. 【Conclusion】 In this case, the development of anti-Jr^a in Jr(a-) mother was stimulated by mother-child serology incompatibility during pregnancy.

【Objective】 To establish an experimental method for detecting phagocytosis of sensitized red blood cells in vitro by flow cytometry. 【Methods】 Mononuclear cells were isolated from the peripheral blood of blood donors and cultured in a cell incubator for 1 hour, and then adherent monocytes were isolated and obtained. Di^b-positive red blood cells (RBCs) were labeled with PKH26 and then sensitized with IgG anti-Di^b. The sensitized RBCs were added to monocytes for in vitro phagocytosis assay. Monocytes were labeled with FITC anti-human CD14, then phagocytosis was measured by flow cytometry, and the phagocytic efficiency was calculated. The method was used to detect the phagocytic efficiency of monocytes on human IgG anti-D sensitized RBCs with different titers. 【Results】 The phagocytic efficiency of monocytes was averaged at 5% (1.2%~7.6%, SD 3.30) versus 81% (71.4%~92.7%, SD 8.65) in the negative versus positive control group, respectively. Phagocytic activity of monocytes mediated by anti-D was correlated with the antibody titer. The phagocytosis efficiency was within 10% when the antibody titer was lower than 32 and increased sharply when the titer was between 32 to 128, it entered a plateau and stabilized at 80% at the titer above 256. 【Conclusion】 A detection platform for detecting phagocytosis-sensitized RBCs in vitro by flow cytometry has been successfully established. It can be used to assess the clinical significance of red blood cell allotype or autologous IgG antibodies.

【Objective】 To establish a high-throughput detection method for ABCG2^*376T allele of Jr(a-), and apply it to the study of the frequency of this allele in the Chinese population. 【Methods】 The specific primers were designed and synthesized, the sample carrying homozygous ABCG2^*376T alleles, obtained in the previous study, was used as the homozygous positive control, and the sample carrying heterozygous allele as the heterozygous positive control. The wild-type sample was used as a negative control, and a high-resolution melting curve(HRM) method for detecting this allele was established. The established method was used to screen DNA samples from blood donors in Guangzhou, and the samples carrying ABCG2^*376T alleles were sequenced to confirm the accuracy of the HRM method. 【Results】 A HRM method, which can detect ABCG2^*376T allele and accurately type homozygotes and heterozygotes at the same time, had been established successfully. Fifteen individuals with heterozygous alleles were screened out of 1 560 blood donors in Guangzhou, while none homozygous allele was detected. 【Conclusion】 The HRM method can be used to accurately screen and type ABCG2^*376T allele. The frequency of this allele in Chinese population is about 0.48%(15/3120).

【Objective】 To explore the identification method and characteristics of anti-IFC against Cromer blood group. 【Methods】 ABO blood group was identified by tube method, and Rh blood group was identified by Rh typing card. Irregular antibody screening and antibody identification were carried out using microtube column agglutination technology(MCAT). The serum of the patient reacted with panel cells treated with antitrypsin, trypsin and bromelain respectively to determine the specificity of the antibody. The serum was inhibited with pure CD55 protein for antibody identification. The DAF, the regulatory gene of Cromer blood group system, was amplified and sequenced. The expression of CD55 on cell membrane was analyzed by flow cytometry. 【Results】 The patient′s blood type was B, CcDEe. The patient′s serum reacted with all the untreated panel red cells, bromelain-treated red cells, trpsin-treated red cells, and DTT treated red cells.It was negative with chymotypsin-treated cells and could be neutralized by CD55 protein. No mutation was found by DAF sequencing. The expression of CD55 on patient′s cell membrane was deficient. 【Conclusion】 This high frequency antibody was identified as anti-IFC. The transient depression in CD55 protein may due to the patient′s GI system abnormalities.

Objectives:To explore health-related quality of life (HRQoL) and identify its associated variables in Chinese patients with Philadelphia-negative myeloproliferative neoplasms (MPNs) .Methods:In this cross-sectional study, anonymous questionnaires were distributed to adult patients with MPNs to assess symptom burden measured by MPN-10 and HRQoL measured by Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36) and the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire Core 30 (EORTC QLQ-C30) .Results:The data from 1405 respondents with MPNs, including 645 (45.9%) with essential thrombocythemia (ET) , 297 (21.1%) with polycythemia vera (PV) , and 463 (33.0%) with myelofibrosis (MF) , were analyzed. 646 (46.0%) respondents were male. The median age was 56 (range, 18-99) years. The mean MPN-10 scores were 13.0±12.7, 15.0±14.7, and 21.0±16.6 ( P<0.001) , and the physical component summary (PCS) and mental component summary (MCS) scores were 48.0±8.5, 47.0±9.0, and 42.0±10.0 ( P<0.001) and 51.0±11.0, 50.0±10.8, and 49.0±11.1 ( P=0.002) for respondents with ET, PV, and MF, respectively. Respondents with MF reported the lowest score of physical functioning, role functioning, emotional functioning, cognitive functioning, social function, and global health status (all P<0.01) and the highest score of fatigue, pain, dyspnea, appetite loss, diarrhea, and financial problems (all P<0.05) in EORTC QLQ-C30. Multivariate analyses revealed that higher MPN-10 scores were significantly associated with lower PCS (-0.220 to -0.277, P<0.001) and MCS (-0.244 to -0.329, P<0.001) scores; increasing age (-1.923 to -4.869; all P<0.05) , lower PCS score. Additionally, comorbidity (ies) , symptom at diagnosis, splenomegaly, anemia, unknown driver gene, and higher annual out-of-pocket cost were significantly associated with lower PCS and/or MCS scores. However, age ≥ 60 years, urban household registration, concomitant medication, and receiving ruxolitinib therapy in respondents with MF were associated with higher MCS scores. Weak correlations were found between MPN-10 score (except the subscale of appetite loss and constipation) and EORTC QLQ-C30 score in majority of subscales in respondents with ET (| r| = 0.193-0.457, all P<0.001) , PV (| r| = 0.192-0.529, all P<0.01) , and MF (| r| = 0.180-0.488, all P<0.001) , respectively. Conclusions:HRQoL in patients with MPN was significantly reduced, especially in patients with MF. Sociodemographic and clinical variables were significantly associated with the HRQoL in patients with MPNs.

Objective:To observe the changes of foveal avascular zone (FAZ) size before and after surgery in idiopathic macular epiretinal membrane (IMEM) eyes and analyze the correlation of FAZ with metamorphopsia.Methods:A retrospective case series study. From August 2016 to October 2017, 42 eyes of 38 patients affected with IMEM diagnosed in Central Theater Command General Hospital of Chinese People's Liberation Army were enrolled in this study. All the patients underwent a 25G pars plana vitrectomy (PPV) with IMEM removal and ininternal limiting membrane (ILM) peeling. The BCVA was measured using the international standard visual acuity chart, and the results were converted to the logMAR visual acuity. The severity of metamorphopsia was measured using M-charts. The FAZ areas were evaluated with OCT angiography in both the superficial and deep capillary plexus layers. The central macular thickness (CMT) were assessed with spectral-domain OCT before and after surgery. The logMAR BCVA was 0.61±0.21. The M-score was 0.66±0.38. The CMT of fovea was 337.71±57.63 μm. The FAZ areas in superficial and deep capillary plexus were 0.113±0.037 mm 2 and 0.202±0.03 mm 2, respectively. The differences in BCVA, M-score and FAZ area before and 1, 3, 6, 12 months after surgery were analyzed by ANOVA. The Spearman rank correlation analysis was performed to investigate the relationship between FAZ areas, visual acuity and metamorphopsia. Results:At 12 month after surgery, the FAZ areas in superficial and deep capillary plexus were 0.146±0.021 mm 2 and 0.240±0.019 mm 2, respectively. Compared with baseline, the the FAZ areas in superficial and deep capillary plexus after surgery significantly increased ( F=8.484, 14.346; P<0.001,<0.001). The postoperative logMAR BCVA 0.47±0.19, M-score 0.12±0.22 and CMT 270.60±33.27 μm were significantly improved compared with baseline ( F=5.044, 17.763, 13.545; P=0.001, <0.001, <0.001). The preoperative FAZ area in superficial capillary plexus correlated negatively with preoperative M-score ( r=-0.816, P<0.001); the preoperative FAZ area in deep capillary plexus correlated negatively with preoperative BCVA and M-score ( r=-0.422, -0.882; P=0.005,<0.001). The postoperative FAZ area in superficial capillary plexus correlated negatively with preoperative and postoperative M-score ( r=-0.791,-0.716; P<0.001,<0.001). The postoperative FAZ area in deep capillary plexus correlated negatively with BCVA and preoperative and postoperative M-score ( r=-0.343, -0.330, -0.732, -0.694; P=0.026, 0.033,<0.001,<0.001). Conclusions:PPV with ILM peeling can effectively restore the FAZ areas in superficial and deep capillary plexus, improve the visual acuity and metamorphopsia in patients with IMEM. Both superficial and deep plexus FAZ areas correlated negatively with metamorphopsia, and deep plexus FAZ area also correlated negatively with BCVA.

Objective:To evaluate the therapeutic efficacy of vitrectomy with internal limiting membrane (ILM) peeling and subretinal injection of balance salt solution (BSS) for refractory diabetic macular edema (DME).Methods:A retrospective case series study. From November 2017 to August 2018, 24 eyes of 19 patients affected with DME resistant to anti-VEGF therapy [central macualar thickness (CMT) more than 275 μm despite undergoing anti-VEGF therapy at least 3 times] in Ophtalmology Department of Central Theater Command General Hospital of Chinese People's Liberation Army were enrolled in this study. All the patients underwent 25G pars plana vitrectomy with ILM peeling and subretinal injection of BSS. The BCVA was measured using the international standard visual acuity chart, and the results were converted to the logMAR visual acuity. The CMT and the macular volume (MV) were assessed with swept-source optical coherence tomography at baseline and each month postoperatively. The differences in BCVA, CMT and MV before and after surgery were analyzed.Results:The mean BCVA was 0.74±0.29 at baseline, which increased significantly to 0.62±0.28, 0.56±0.25, 0.47±0.26, 0.46±0.23 at 2 weeks, 1 month, 3 months and 6 months after treatment respectively ( F=4.828, P=0.001). At 6 months, BCVA improved by more than 0.3 logMAR units in 16 eyes (66.7%). The mean CMT was 554.58±102.86 μm at baseline, which reduced to 338.17±58.09 μm,299.42±52.66 μm, 275.75±41.24 μm and 270.96±38.33 μm at 2 weeks, 1 month, 3 months and 6 months after treatment respectively ( F=84.867, P<0.001). The mean MV was 13.01±0.88 mm 3 at baseline, which decreased to 11.50±0.73 mm 3, 11.00±0.74 mm 3, 10.68±0.61 mm 3 and 10.52±0.56 mm 3 at 2 weeks, 1 month, 3 months and 6 months after treatment respectively ( F=47.364, P<0.001). Macular edema recurred in 5 eyes(20.8%) 6 months after surgery. No severe systemic or ocular side effect was reported during the follow-up. Conclusions:25G vitrectomy with ILM peeling and subretinal injection of BSS for refractory DME can improve the visual acuity, facilitate a rapid resolution of macular edema.

OBJECTIVE: To explore the correlation between special A/O genotype and the O phenotype. METHODS: Group O samples with partially reduced or lack of isoagglutinins were collected to determinate the ABO genotype with a PCR-sequence specific primer (PCR-SSP) assay. Seven samples with A/O genotype were selected for further study. Serological tests including forward and reverse typing, H antigen determination and adsorption/elution were carried out with a tube method. Genomic DNA was genotyped by amplifying and sequencing of the coding regions of exons 1 to 7 of the ABO gene. RESULTS: Seven samples were serotyped as group O by the forward typing test. However, reduced anti-A activity was found in 5 samples by the reverse typing test, reduced anti-A and anti-B activities were found in 1 sample, and no anti-A isoagglutinin activity was found with 1 sample. H antigen was determined in all samples by routine serologic method. Neither anti-A nor anti-B was eluted from red cells derived from all samples. Three samples were genotyped as Ael02/O02, whilst the remainders were Ael02/O13, Ael02/O65, Am04/O75, Ael06/O02, respectively. CONCLUSION Special A/O genotype may not express the A antigen, leading to the generation of group O red cells. Reduced or missed anti-A activity is the typical serological feature of this special group of O phenotype, for which ABO*Ael02 and ABO*O02 are the major alleles. Group O individuals with isoagglutinin detection problem should be grouped by serological tests and genomic DNA analysis.
ABO Blood-Group System ; Alleles ; Exons ; Genotype ; Humans ; Phenotype