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Red blood cells (RBCs) of the Asian-type DEL phenotype express few RhD proteins and are typed as serologic RhD-negative (D-) in routine testing. RhD-positive (D+) RBC transfusion for Asian-type DEL patients has been proposed but has not been generally adopted due to a lack of direct evidence regarding its safety and underlying mechanism. We performed a single-arm multicenter clinical trial to document the outcome of D+ RBC transfusion in Asian-type DEL patients; none of the recipients (0/42; 95% confidence interval, 0%-8.40%) developed alloanti-D after a median follow-up of 226 days. We conducted a large retrospective study to detect alloanti-D immunization in 4,045 serologic D- pregnant women throughout China; alloanti-D was found only in true D- individuals (2.63%, 79/3,009), but not in those with Asian-type DEL (0/1,032). We further retrospectively examined 127 serologic D- pregnant women who had developed alloanti-D and found none with Asian-type DEL (0/127). Finally, we analyzed RHD transcripts from Asian-type DEL erythroblasts and examined antigen epitopes expressed by various RHD transcripts in vitro, finding a low abundance of full-length RHD transcripts (0.18% of the total) expressing RhD antigens carrying the entire repertoire of epitopes, which could explain the immune tolerance against D+ RBCs. Our results provide multiple lines of evidence that individuals with Asian-type DEL cannot produce alloanti-D when exposed to D+ RBCs following transfusion or pregnancy. Therefore, we recommend considering D+ RBC transfusion and discontinuing anti-D prophylaxis in Asian-type DEL patients, including pregnant women. This clinical trial is registered at www.clinicaltrials.gov as NCT03727230.

The molecular basis of the D variant phenotype in the Chinese differs greatly from that of the Caucasian. Adapting a specific D typing strategy to the spectrum of prevalent RHD variant alleles is necessary.Blood samples with ambiguous D phenotypes were collected in the Southern Chinese population. A special three-step typing strategy was applied. First, the common DVI type 3 was identified from epitope profiles of D antigen. Then, another common weak D type 15 (RHD*845A) was identified by epitope profiles of D antigen and Sanger sequencing of RHD exon 6. Finally, the remaining D variants were genotyped mainly by Sanger sequencing. For the novel RHD alleles in the coding region and exon-intron junction, in vitro transfection and minigene splicing assays were performed, respectively. The anti-D investigation was performed.DVI type 3 (65/253, 25.7%) and weak D type 15 (62/253, 24.5%) were common Chinese D variants, and RHD*960A, DFR, RHD*weak D type 25, 72, and 136 were frequent variant RHD alleles. Besides, twenty-two sporadic and seven novel RHD alleles (RHD*188A; RHD*688C; RHD*782 T; RHD*1181C; RHD*165 T, 993A; RHD*148 + 3G T and RHD*1227 + 5G C) were identified. The deleterious effect of the novel RHD alleles on D antigen or mRNA expression was confirmed. Anti-D was detected in two DVI type 3 pregnant women.The three-step typing strategy provides an effective approach for Chinese D variant typing. It can be anticipated that commercially available RHD genotyping kits have limitations for testing Chinese D variants, as some of the frequent variants are not interrogated.

Background The hybrid glycophorins of MNS blood group system express a series of low incidence antigens including Mia , which are commonly found in Southeast Asian populations. In this study, the molecular basis of Mia -positive hybrid glycophorins was firstly clarified in the Chinese Southern Han population. RNA transcripts of GYPB gene in the homozygous GP.Mur individuals were also analyzed. Study design and methods DNAs were extracted from the whole blood samples of 111 Mia -positive donors. Then, high-resolution melting (HRM) analysis for GYP(B-A-B) was used to analyze the genotypes. Sequencing of GYPB pseudoexon 3 was conducted in the samples with variant melting curves. TA-cloning and subsequent sequencing of GYPA exons 2-4 were performed in the Mia -positive samples with normal GYPB/GYPB genotype by HRM. The transcript analysis of GYPB was conducted in homozygous GP.Mur and wild-type glycophorin B (GPB) individuals using RNA extracted from the cultured erythroblast. Results The heterozygous GYP*Mur/GYPB (n = 101), homozygous GYP*Mur/GYP*Mur (n = 7) including one novel GYP*Mur allele with an extra GYPA/GYPE specific nucleotide substitution (c.229+110A>T), heterozygous GYP*Bun/GYPB (n = 1) and GYP*Vw/GYPA (n = 2) with two novel GYP*Vw alleles were identified. RNA transcript analysis revealed multiple transcripts of GYPB existing in both homozygous GP.Mur and normal GPB individuals. Conclusion The results showed the genetic diversity of hybrid glycophorins in the Chinese population. Besides, the successful analysis of GYPB transcripts indicates that the cultured erythroblast is a good source for RNA transcript analysis for the protein only expressed on the red blood cells.

Objective To investigate the effect of over-expression of nudix hydrolase 21 (NUDT21) on the proliferation of colon cancer HCT-116 cells and its mechanism. Methods The NUDT21 over-expression plasmid was constructed by Gibson assembly. The colony formation assay and CCK-8 assay were used to detect cell proliferation. The cell cycle of HCT-116 cells was detected by flow cytometry. Western blot was performed to detect the expressions of P53, cyclin-dependent kinase 2 (CDK2), phosphorylated retinoblastoma protein at serine 780 (p-Rb-Ser780), and p-Rb-Ser608. Results The sequencing results showed that the NUDT21 over-expression plasmid was successfully constructed. After the NUDT21 over-expression plasmid was transfected into HCT-116 cells, the expressions of NUDT21 mRNA and protein in the cells were significantly increased. The over-expression of NUDT21 inhibited the proliferation of HCT-116 cells and arrested the cell cycle in G0/G1 phase. The expressions of CDK2, p-Rb-Ser608, and p-Rb-Ser780 proteins decreased while the expression of P53 protein increased. Conclusion Over-expression of NUDT21 inhibits the proliferation of HCT-116 cells by blocking P53/CDK2/Rb signal pathway.

Background The distribution of DI1/DI2 antigens of the Diego blood group system is polymorphic in Mongoloid populations and the corresponding alloantibodies are clinically significant. Here a novel DI variant was found by donor screening, and the effect of the novel and previously reported mutations on expression of DI1/DI2 antigens and Band 3 protein was explored. Study design and methods DNA samples of 1150 Chinese donors were collected. DI*01/DI*02 genotyping was determined by Sanger sequencing. For the carrier of novel allele, the expression of Band 3 and DI1/DI2 antigens on red blood cells (RBCs) was detected by Western blot and flow cytometry, respectively. in vitro expression studies were conducted by transfecting the mutant (including the novel and three reported DI*02(2534T), DI*02(2358_2359insCAC), and DI*02(2572T) alleles) or wild-type DI*02 constructs into HEK 293T cells, the expression of Band 3 and DI1/DI2 antigens was analyzed. Results A novel heterozygous mutation (c.2558C>T, p.Thr853Met), which is located near the DI1/DI2 polymorphism site (c.2561T>C), was identified in a donor with DI:-1,2 phenotype. Reduced expression of DI2 antigen was observed on the RBCs, while weakened expression of Band 3 and absence of DI2 antigen were detected in cells transfected with the mutant DI*02(2558T) construct. In addition, absent or decreased expression of Band 3 and DI2 antigen was also detected in cells transfected with three reported mutant constructs. Conclusion The novel DI*02(2558T) allele and three previously described DI mutations can affect the expression of Band 3 protein and/or DI2 antigen and/or interfere with DI*01/DI*02 genotyping result.

"Asia type" DEL red blood cells (RBCs) express a very weak D antigen and cannot be detected by routine RhD typing. Thus, it is routinely typed as D-negative (D-) blood group and transfused to D- recipients. Here we described a case of secondary alloanti-D immunization that was associated with transfusion of DEL RBCs to D- recipients and was initially considered as primary alloanti-D immunization.A 44-year-old D- woman (G2P2) with adenomyosis and anemia underwent transabdominal hysterectomy. She received four units of D- RBCs before operation. Before transfusion, the alloantibody screening test was negative. Four days after the first transfusion, she needed another RBC transfusion. Unexpectedly, the routine pre-transfusion alloantibody screening test became positive and anti-D (titer, 128-fold) was identified, indicating an alloanti-D immunization. The anti-D developed four days after the first transfusion was unexplained, so alloantibody identification was performed on the sample collected before the first transfusion, and weak anti-D combined with anti-E, which was not detectable during the previous routine pre-transfusion alloantibody screening test with non-enzyme-treated screening cells, was identified using bromelain-treated panel cells. The remaining blood samples of first transfusion in bag tails from two donors were collected for RHD genotyping analysis. One donor was later identified as "Asia type" DEL having RHD* 1227 A/01 N.01 genotype.Caution should be applied when we conclude that transfusion of "Asia type" DEL RBCs to true D- recipients could induce primary alloanti-D immunization, especially if the short time interval between transfusion and detection of anti-D is observed.

Fetal and neonatal alloimmune thrombocytopenia (FNAIT) is a bleeding disease that can cause fetal hydrops, a rare but life-threatening condition in which abnormal amounts of fluid accumulate in one or two areas of the fetus's body. A case of FNAIT with fetal hydrops caused by anti-HPA-15b antibodies was involved in this study, as we investigated whether or not anti-HPA-15b antibodies can induce endothelial angiogenesis and apoptosis.The monoclonal antibody immobilization of platelet antigens assay (MAIPA) was used to identify anti-HPA-15b antibodies. The three groups in Tube formation and apoptosis assays were the PBS group, the AB serum IgG group, and the anti-HPA-15b serum IgG group, all reacted with HPA-15bb HUVEC.The presence of anti-HPA-15b antibodies was found in this case by MAIPA assay. The OD values are 0.33 and 0.21, reacted with HPA-15bb and HPA-15ab platelets, respectively (cutoff OD value = 0.2). Quantitative analysis revealed that the length of capillary-like tube induced by anti-HPA-15b antibodies was significantly decreased over that of AB serum IgG (*p = 0.0005), but weaker than when incubated with thrombin (**p = 0.0009). The apoptosis results show a significantly increased number of apoptotic endothelial cells in the anti-HPA-15b antibody IgG group when compared with the PBS and AB serum IgG groups (*p 0.0001, **p 0.0001). In addition, there is no statistical difference between the PBS and AB serum groups.Anti-HPA-15b antibodies can inhibit angiogenesis and induce apoptosis. This may associate with hydrops fetalis (HF), or fetal hydrops of FNAIT.

Background: Arsenide, a kind of pollution widely existing in the environment, and the mechanism of occurrence and development of lung cancer leading by long-term arsenic exposure still needs further research up to present. We investigated how high and low doses of arsenic induce human bronchial epithelial cell transformation and the molecular mechanisms involvedMethods: After exposing human bronchial epithelial cells(Beas-2B) to different doses of sodium arsenite for 24 h, RNA-seq analysis was performed to detect arsenic-regulated genes; for cell viability and status and for cell cycle changes and mitochondrial function; long-term exposure to 1μM sodium arsenite after 20 consecutive passages for cell cycle changes and mitochondrial function.Results: In the study, human bronchial epithelial cells were exposed to varying high-dose sodium arsenite in short term or treated with low dose in a long term, and the data showed that both short and long terms treatment promoted G1/S transition of Beas-2B cells, inducing significant increases of expression of AKAP95, cyclin D1, cyclin D2 and cyclin E1. However, silencing AKAP95 by treating cells with siAKAP95 showed a protective function that inhibited G1/S transition, suggesting that regulatory mechanism of AKAP95 on cell cycle during cell malignant transformation induced by sodium arsenite. In addition, we also noticed some mitochondrial dysfunctions occurred during sodium arsenite exposure, including the disappearance of mitochondrial double membrane structure, the formation of vacuole structure, the decrease of mitochondrial membrane potential, the change of expression levels of mitochondrial related proteins Tim22, Tim23, Tom40 and OPA1, the release of lactate dehydrogenase (LDH) and the decrease of reduced glutathione.Beas-2B cells, which exposed to low-dose sodium arsenite for a long term, were subcultured for 20 generations, and it was found that the exposure time was positively proportional to the growth and migration rate of cells. The exposed cells were used in tumor bearing transplantation experiment (mice), and results showed that the longer the exposure time, the faster the tumor volume growth rate of As-Beas-2B cells. Tumor tissues were taken out for HE staining, and it was found that the cell morphology changed and the volume increasedConclusion: high and low doses of sodium arsenite induced malignant transformation of human bronchial epithelial cells by promoting G1/S turnover through AKAP95 with associated cyclins, accompanied by the development of mitochondrial dysfunction.

The transfusion of D-negative red blood cells (RBCs) to D-negative patients has been widely adopted to prevent anti-D alloimmunization, especially in women of childbearing age. Still, transfusion of D-positive RBCs to D-negative recipients is occasionally inevitable in practice, and the resulting incidence of anti-D in different D-negative groups of patients has not been well summarized.We searched the relevant literature using PubMed, Cochrane Library, and Embase databases from inception date to 30 September 2021. We looked for studies of anti-D occurring in D-negative recipients who received D-positive RBC transfusions. The anti-D incidence was summarized with 95% confidence intervals (CIs). Data with similar characteristics were combined using a random-effects model.About 42 studies (2226 cases), which found anti-D, the exact volume of D-positive RBC transfused, and the follow-up time for anti-D detection, met the inclusion criteria. The pooled anti-D incidence was 64% (95% CI, range 55%-74%) in volunteers receiving small volumes of D-positive RBCs, 84% (95% CI, 74%-94%) in those receiving whole units, 26% (95% CI, 19%-32%) in mixed patients, 12% (95% CI, 8%-16%) in oncology patients, 27% (95% CI, 13%-40%) in trauma patients, 4% (95% CI, 0%-8%) in immune-compromised transplant patients, and 6% (95% CI, 1%-39%) in those with AIDS.Compared with the high frequency of anti-D in healthy D-negative volunteers given D-positive RBCs, we found a lower rate of anti-D immunization in various D-negative patients and almost none in transplant and AIDS patients.

The Ordovician large-scale gas reservoir in the Tazhong III region (burial depth > 6000 m) is a typical deep strata gas reservoir in the Tarim Basin of NW China. The origin, migration and accumulation of the Ordovician natural gas in the Tazhong III region were investigated by integrating geological, geophysical, and geochemical parameters. The parameters that were successfully analyzed included the gas compositions and carbon isotope ratios, production data, burial and geothermal histories, oil compositions and properties, together with the diamantane index. The results indicated that the Ordovician natural gas in the Tazhong III region originated from Cambrian-Ordovician paleo-oil cracking, accompanied by a large amount of H2S generated from thermochemical sulfate reduction (TSR). When the oil cracking gas migrated vertically into the Ordovician reservoir through the Tazhong I fault and strike-slip faults, it was preferentially charged in the pinnate fault zones of strike-slip faults with high porosity and permeability, leading to the enrichment of natural gas and an abnormal distribution of the compositions and properties of hydrocarbon. Then, oil cracking gas intruded into and altered the previous oil reservoir from northeast to southwest through the strike-slip faults, unconformities and pore-fracture reservoir belt, leading to the differential accumulation of hydrocarbon in districts of the Tazhong III region. The Ordovician natural gas migration-accumulation model in the Tazhong III region indicates a great exploration potential in the deep or super-deep strata of the Tarim Basin. Further exploration activities in the Ordovician reservoir of the Tarim Basin should be conducted in the regions where Cambrian-Ordovician source rocks are well developed, and the stress fracture zones-related deep fault should be emphasized.

Arsenic is a widely existing pollutant in the environment, but the mechanism of occurrence and development of lung cancer by long-term arsenic exposure needs to be elucidated further. How the high and low doses of arsenic induce human bronchial epithelial cell transformation is yet to be elucidated. In the present study, human bronchial epithelial cells were exposed to varying high-dose sodium arsenite (NaAsO

Objectives To screen RhCE variants in the Chinese Southern Han donors for molecular genetic analysis. Background More than hundreds of RhCE variant alleles have been described to resulting in weak and/or partial expression of RhCE antigens, generation of low-prevalence antigens and/or absence of a high-prevalence antigen of Rh system, which mainly reported in the people of African origin. In this study, the serological screening and molecular genetic analysis of RhCE variants were performed in the Chinese Southern Han donors. Methods The blood samples of E(+) donors were preliminarily collected. Then, RhCE antigens of the E(+) samples were further typed by using two sets of monoclonal anti-C, anti-c, anti-e and another anti-E. When weak expression of RhCE antigens was found, direct sequencing for 10 exons of RHCE gene, RH genotyping analysis by using multiplex ligation-dependent probe amplification, flow cytometric analysis and even cDNA sequencing were performed. Results A total of 4487 E(+) samples were collected and four samples with weak expression of antigens were detected. RHCE*Ce375G and RHCE*Ce667T variant alleles were identified in two samples with weak expression of e antigen, respectively. But no variant alleles were found in another two samples with weak expression of C antigen. Conclusion The variant RHCE*Ce375G validated by mRNA sequencing and the deduced RHCE*Ce667T alleles were firstly identified in the Chinese population. The DCE haplotype might account for the weak expression of C antigen in two donors.

The hybrid glycophorins of MNS blood group system express a series of low incidence antigens including MiDNAs were extracted from the whole blood samples of 111 MiThe heterozygous GYP*Mur/GYPB (n = 101), homozygous GYP*Mur/GYP*Mur (n = 7) including one novel GYP*Mur allele with an extra GYPA/GYPE specific nucleotide substitution (c.229+110AT), heterozygous GYP*Bun/GYPB (n = 1) and GYP*Vw/GYPA (n = 2) with two novel GYP*Vw alleles were identified. RNA transcript analysis revealed multiple transcripts of GYPB existing in both homozygous GP.Mur and normal GPB individuals.The results showed the genetic diversity of hybrid glycophorins in the Chinese population. Besides, the successful analysis of GYPB transcripts indicates that the cultured erythroblast is a good source for RNA transcript analysis for the protein only expressed on the red blood cells.

Purpose: Cx43 (connexin 43) has been found to inhibit cell cycle of tumor cells. No nuclear localization sequence of Cx43 was found but the protein has been found in nucleus. The process remains unclear. We are aiming at providing the model that AKAP95 (A-kinase anchoring protein) binds to and transports Cx43 into nucleus of lung cancer cells.Methods: Lung cancer cells were arrested at preliminary stage (P), middle and late stage (M) of G1 phase and restriction point (R). Immunocoprecipitation (Co-IP) and Western blot (WB) were used to analyze expression and binding of AKAP95, Cx43, cyclin D1 and cyclin E1 at different stages of lung cancer cells during G1 phase. Confocal laser scanning microscopy (CLSM) was used to detect the location of proteins in arrested cells. Transmission electron microscope (TEM) with higher resolution was used to detect binding/aggregating and location of the above four proteins at subcellular level.Results: 1) AKAP95 transports Cx43 into the nucleus through the nuclear pore mainly at R stage. Some AKAP95 and Cx43 form protein aggregates during the process. 2) Cyclin E1 was found to be bind/aggregate with AKAP95-Cx43 complex and involved in the nuclear transport of Cx43 mainly at R stage. 3)Cyclin D1 was found to bind with AKAP95 and Cx43 respectively but not to be involved in their aggregating and nuclear transport of Cx43.Conclusions: 1) AKAP95 and Cx43 bind/aggregate to each other during G1 phase, forming AKAP95-Cx43 complexes/aggregates. After their binding/aggregating, AKAP95 targets nuclear pore and matrix, transporting Cx43 into nucleus. Formed AKAP95-Cx43 complexes/ aggregates do not dissociate in nucleus. The process can be summarized as ‘forming AKAP95-Cx43 complexes/aggregates; transporting Cx43 into nucleus; functioning remaining binding/aggregating’. 2) Cyclin D1 was only found to bind to AKAP95 and Cx43 respectively and function in cytoplasm while cyclin E1 was involved in the binding/aggregation of AKAP95 and Cx43, as well as the nuclear transport of Cx43.

BackgroundAKAP95(A-kinase anchoring protein) and Cx43 (connexin 43) express abnormally in lung cancer cells. As potential tumor therapeutic targets, specific process of bindings and dynamic changes of Cx43-AKAP95 complex in lung cancer cells may guide further treatments and detections of lung cancer. However, the process remains unclear now. We are aiming at investigating the dynamic changes of expression, localization, and binding of AKAP95 and Cx43, as well as their interaction with cyclin D1 and cyclin E1 in lung cancer cells during G1 phase.MethodsA549 and Beas-2B cells were arrested at preliminary stage(P), middle stage(M) and restriction point(R) of G1 phase and Western blot(WB), Confocal laser scanning microscopy (CLSM) and Transmission electron microscope (TEM) were used in our study to detect proteins to provide further evidence of the correlation of these proteins.Results: 1) AKAP95 carries Cx43 into the nucleus through the nuclear pore by binding to it during G1 phase. Some AKAP95 and Cx43 can aggregate and form larger protein aggregates. The process happens mainly during R stage. 2) AKAP95 and Cx43 mainly bind to cyclin D1 during P and M stage while bind to cyclin E1 during R stage respectively. Complexes of AKAP95-cyclin D1 and Cx43-cyclin D1 cannot enter nucleus while AKAP95-cyclin E1 and Cx43-cyclin E1 can.Conclusions1) Binding process of AKAP95-Cx43 complexes/ aggregates can be summarize as ‘bind and aggregate -target and enter nucleus- keep binding/aggregating in nucleus’. 2) Cyclin E1 was involved in the binding/aggregation and nuclear entry of AKAP95 and Cx43 while cyclin D1 only binds to them respectively in the cytoplasm.

Background Haemolytic disease of the foetus and newborn (HDFN) is the most common aetiology of haemolytic anaemia and hyperbilirubinaemia in foetuses and neonates. Studies on the distribution of antibodies that cause haemolytic disease of the foetus (HDF) in China are limited, and the effects of multiple antibodies on the severity of HDF need further evaluation. Methods An observational cohort study from January 2005 to December 2019 was conducted in two hospitals affiliated with Sun Yat-sen University. Maternal red cell alloimmunization was identified by the Guangzhou Blood Centre. In total, 268 pregnant woman-foetus pairs were divided into four groups according to the type of maternal alloantibodies: anti-D, anti-D combined with other antibodies, other single-antibody and other multiple antibodies. The obstetric history, antibody characteristics, incidence of severe HDF and foetal outcomes were collected and compared. Logistic regression analysis of the risk factors for HDF and survival analysis of the severe HDF-free interval were conducted. Results Anti-D was the most common cause of HDF, followed by anti-M. No anti-K- or isolated anti-c-associated HDF was found. The incidence of severe HDF was higher in the group with anti-D combined with other antibodies than in the group with anti-D alone (P = 0.025), but no significant difference was found in haemoglobin level and reticulocyte count in the anaemic foetuses between these two groups. Foetuses in the other single-antibody group had a lower reticulocyte count (P = 0.007), more IUTs (P = 0.007) and an earlier onset of severe HDF (P = 0.012). The maximum antibody titre was significantly lower in the other single-antibody group than in the anti-D group (P P P P = 0.042) were independent risk factors for HDF. A higher reticulocyte count (P = 0.041) was an independent risk factor for severe HDF in anaemia foetuses affected by Rh(D) alloimmunization. Conclusions The distribution of HDF-associated antibodies in China is different from that in Western countries. Other single non-Rh(D) antibodies could increase the risk of HDF, and anti-D combined with other antibodies would not influence the severity of foetal anaemia compared with anti-D alone.

Data masking is the core technology to eliminate privacy sensitive information in finance, power and other industries. FPE (Format Preserving Encryption) is used for data desensitization due to the reason that its ciphertext and plaintext are in the same message space. Both the ciphertext and plaintext have the same format characteristics. Based on the traditional format-preserving encryption desensitization scheme, this paper proposes a general model for the application of identity-based format pre-encryption (IB-FPE) in data desensitization technology. Firstly, the solution preserves the traditional format retention encryption scheme and maintains data integrity efficiently. Secondly, by improving the key generation function, the identity information is associated with the master key, thereby locating and limiting the damage caused by the key exposure to the format retention encryption. Finally, by increasing the processing opinion of segmented data, the efficiency of the format-preserving encryption desensitization scheme is improved and the data desensitization process of the specific example is analyzed, and the security and efficiency of the scheme are analyzed.

Background The molecular events resulting in a weak D phenotype include missense mutations, in-frame insertion, or deletion mutations of the RHD gene and hybrid RHD-CE-D hybrid alleles. Mutations in genes encoding the proteins that are required for proper membrane expression of Rh proteins, such as RhAG and ankyrin 1, can lead to absent or weakened expression of Rh antigens. Study design and methods Blood sample from a Chinese blood donor with a serological weak D phenotype was collected. RhAG antigen expression, RhD, and RhCE phenotypes were determined. Analysis of the RHD and RHCE genotypes by RH multiplex ligation-dependent probe amplification (MLPA), Sanger sequencing of the RHD exons, and next-generation sequencing (NGS) of the RHAG and ANK1 exons were performed. Expression studies in vitro were conducted by lentivirally transducing the mutant RHAG*572A or wild-type RHAG, in combination with either RHD or RHCE constructs, into HEK 293 T cells. The expression of RhAG, RhD, and RhCE antigens was analyzed by flow cytometry. Results Serological weak D and normal C + c- E- e + phenotypes, normal CCDDee genotype determined by RH-MLPA, and normal sequence of the RHD gene by Sanger sequencing were demonstrated. A homozygous variant (c.572G > A, p.Arg191Gln) of the RHAG gene was revealed by NGS analysis. Normal RhAG, weak RhD, and normal RhCE antigens were detected in cells transduced with the mutant RHAG*572A, the mutant RHAG*572A and RHD, and the mutant RHAG*572A and RHCE constructs, respectively. Conclusion The homozygous presence of RHAG*572A allele results in weak D expression. It does not affect RhCE expression.

The accumulation mechanism of hydrocarbons in the Yingmai 2 Oilfield of the Tabei Uplift in the Tarim Basin is not well understood. Here, the characteristics and formation of Paleozoic faults in the Yingmai 2 Oilfield are studied with new 3D seismic data and regional geologic data. Inclusion features and geochemical parameters are used to investigate the faults controlling the oil and gas compositions. The results indicate that three fault systems developed in the Yingmai 2 Oilfield: a NNW-trending strike-slip fault system that formed in the middle Caledonian, a NW-trending strike-slip fault system that formed in the Caledonian–late Hercynian and a NE-trending thrust fault system that formed in the late Hercynian–Indosinian, which was associated with volcanic activity. The two strike-slip fault systems were important early hydrocarbon migration pathways in the late Caledonian–early Hercynian, late Hercynian, and Himalayan accumulation periods. The NE-trending thrust faults were sealed by volcanics, and divided the originally uniform oilfield into two accumulation systems during the Himalayan. This finding is supported by the differences in the compositions of the oils and gases in the eastern (YM area) and the western (YG area) Yingmai 2 Oilfield. The oil density, iC4/nC4, Ph/nC18, and V/Ni ratios of the oils in the reservoirs in the YM area are lower than those in the YG area, whereas the TMNr, TeMNr, 4-1/DBT, Pr/Ph, C29-diasterane/C29-regular sterane ratios, H2S content, and the content of Himalayan hydrocarbon inclusions are higher. The hydrocarbon accumulation in the Yingmai 2 Oilfield is attributed to multiple episodes of hydrocarbon charging and suggests that the YM area has greater exploration potential than the YG area.

Hydrocarbon migration mechanism in deep Ordovician reservoir of the Halahatang area, Tarim Basin, is investigated based on the analysis of the geological setting and geochemical parameters. The parameters successfully analyzed include oil–gas properties, crude oil maturity [Ts/(Ts + Tm) ratio], gas carbon isotopic ratios, and trace elements (B, V, Ni, and Cu) in crude oil. The results show that hydrocarbons migrate into the Ordovician of the Halahatang area from the Manjiaer Depression and the Halahatang area itself through unconformities and faults. The overall hydrocarbon migration orientation in the area is generally from south to north. The oil and gas migration process determine parameters vary regularly from south to north as follows: oil density, V/Ni ratio, Ni/Cu ratio, and concentrations of V, Ni and (Ni + B) in crude oil regularly increase, whereas Ts/(Ts + Tm), gas/oil ratio, and dryness coefficient gradually decrease, and methane show a trend toward depletion of heavy carbon isotopes. Additionally, there exist obvious anomalies of geochemical parameter values in the Repu3 well block, which indicates that a local fault intersection could provide a favorable hydrocarbon filling point. Based on these findings, further exploration activities in the deep Ordovician reservoir of the Halahatang area should be conducted in the southern part of the Halahatang area along the oil charging pathways, and fault intersection zones should be emphasized.

BACKGROUND MNS hybrid GP(B-A-B) glycophorins are more commonly found in Southeast Asians and alloantibodies to antigens they carry are clinically significant. Detection of hybrid glycophorins by serologic techniques is limited due to lack of commercial reagents. In this study, a genotyping method for GP(B-A-B) hybrid glycophorins based on high-resolution melting (HRM) analysis was applied for genotyping analysis in the Chinese Southern Han population. STUDY DESIGN AND METHODS DNA samples from 3104 Chinese Southern Han blood donors were collected. GYP(B-A-B) genotypes were analyzed by HRM assay. Parts of samples (n = 106) were also tested by multiplex ligation-dependent probe amplification (MLPA) assay. Direct sequencing was conducted in samples with variant melting curve profiles. RESULTS A total of five GYP(B-A-B) genotypes (201/3104, 6.5%) were identified, which were GYP*Mur heterozygote (n = 194), GYP*Mur homozygote (n = 3), GYP*Bun heterozygote (n = 2), GYP*HF heterozygote (n = 1), and a novel GYP(B-A-B) hybrid allele (n = 1). Genotyping results for GYP*Mur and wild-type GYPB samples obtained by HRM were consistent with MLPA, while GYP*Bun and GYP*HF heterozygote identified by HRM could only be identified to have one copy of 5' inactive splice site of GYPB Pseudoexon 3 by MLPA. In addition, 10 single-nucleotide polymorphisms (SNPs) including four known and six novel SNPs were identified in 31 samples. One sample was identified carrying both GYP*Mur and GYP*Sch alleles. CONCLUSION The HRM assay could distinguish the GYP(B-A-B) hybrid alleles successfully. Polymorphisms identified within the GYPB gene should be taken into consideration when developing GYP(B-A-B) genotyping kits for the Chinese population.

Deep sour gases are found in the Ordovician carbonate reservoir of the Tazhong Uplift in the Tarim Basin, northwest China. All sour gases in this field are dominated by gas hydrocarbons (C1–C4) with varying amounts of H2S. Most of the gases in the eastern Tazhong are characterized by high gas dryness coefficients (>0.95), 13C-enriched methane (δ13C1 ranging from −44.6‰ to −35.8‰), and small difference between values of the δ13C2 and δ13C1 (δ13C2-1). In contrast, the gases in the western Tazhong are characterized by relatively low gas dryness coefficients (mostly

The Ordovician hydrocarbon migration and accumulation of the Tazhong Uplift in the Tarim Basin, northwest China, is investigated from the perspective of geological and geochemical analysis. Geochem...

To explore the correlation between special A/O genotype and the O phenotype.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.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.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.

We investigate the dynamics and bifurcations of SIR epidemic model with horizontal and vertical transmissions and constant treatment rates. It is proved that such SIR epidemic model have up to two positive epidemic equilibria and has no positive disease-free equilibria. We find all the ranges of the parameters involved in the model under which the equilibria of the model are positive. By using the qualitative theory of planar systems and the normal form theory, the phase portraits of each equilibria are obtained. We show that the equilibria of the epidemic system can be saddles, stable nodes, stable or unstable focuses, weak centers or cusps. We prove that the system has the Bogdanov-Takens bifurcations, which exhibit saddle-node bifurcations, Hopf bifurcations and homoclinic bifurcations.

The genetic basis for five GP(B-A-B) MNS system hybrid glycophorin blood group antigens results from rearrangement between the homologous GYPA and GYPB genes. Each hybrid glycophorin displays a characteristic profile of antigens. Currently, no commercial serological reagents are currently available to serologically type for these antigens. The aim of this study was to develop a single nucleotide polymorphism (SNP) mapping genotyping technique to allow characterisation of various GYP(B-A-B) hybrid alleles. Matrix-assisted laser desorption/ionisation time-of-flight (MALDI-TOF) mass spectrometry (MS) assays were designed to genotype five GYP(B-A-B) hybrid alleles. Eight nucleotide positions were targeted and incorporated into the SNP mapping protocol. The allelic frequencies were calculated using peak areas. Sanger sequencing was performed to resolve a GYP*Hop 3' breakpoint. Observed allelic peak area ratios either coincided with the expected ratio or were skewed (above or below) from the expected ratio with switching occurring at and after the expected break point to generate characteristic mass spectral plots for each hybrid. Sequencing showed that the GYP*Hop crossover in the intron 3 region, for this example, was identical to that for GYP*Bun reference sequence. An analytical algorithm using MALDI-TOF MS genotyping platform defined GYPA inserts for five GYP(B-A-B) hybrids. The SNP mapping technique described here demonstrates proof of concept that this technology is viable for genotyping hybrid glycophorins, GYP(A-B-A), GYP(A-B) and GYP(B-A), and addresses the gap in current typing technologies.

Background Alloanti-M was once regarded as not clinically significant, with a few exceptions in extremely rare cases. However, an increasing number of cases of severe hemolytic disease of the fetus and newborn (HDFN), resulting in fetal hydrops and recurrent abortion caused by alloanti-M, have been reported mainly in the Asian population. Study design and methods Three pregnant Chinese women with a history of abnormal pregnancy with hydrops fetalis were encountered. During this pregnancy, a series of clinical examinations and an alloantibody identification against RBCs and platelets were conducted. Intrauterine transfusion and postnatal transfusion were then performed in the fetuses. In addition, the HDFN cases caused by alloanti-M reported in different ethnic groups as well as their clinical and serologic features are also summarized. Results Three pregnant women were identified with an M-N+ phenotype and IgM mixed with IgG alloanti-M in serum. Their fetuses were found by ultrasound examination and cord blood testing to have severe anemia. Additionally, an M+N+ phenotype and IgG alloanti-M were detected in the cord blood of the three fetuses with titers ranging from 1:1 to 1:128. Moreover, low reticulocyte counts and negative direct antiglobulin tests were also shown in two of the fetuses. After receiving intrauterine transfusions and postnatal transfusions several times, these three fetuses eventually survived and then healthfully developed in the follow-up tracking. Conclusion Alloanti-M immunization can cause severe HDFN with hyporegenerative anemia, often seen in the Asian population, and suppression of erythropoiesis might account for it.

p53 tumor-suppressor gene is a master transcription factor which controls cell cycle progression and apoptosis. killin was discovered as one of the p53 target genes implicated in S-phase control coupled to cell death. Due to its extreme proximity to pten tumor-suppressor gene on human chromosome 10, changes in epigenetic modification of killin have also been linked to Cowden syndrome as well as other human cancers. Previous studies revealed that Killin is a high-affinity DNA-binding protein with preference to single-stranded DNA, and it inhibits DNA synthesis in vitro and in vivo. Here, co-localization studies of RFP-Killin with either GFP-PCNA or endogenous single-stranded DNA binding protein RPA during S-phase show that Killin always adopts a mutually exclusive punctuated nuclear expression pattern with the 2 accessory proteins in DNA replication. In contrast, when cells are not in S-phase, RFP-Killin largely congregates in the nucleolus where rRNA transcription normally occurs. Both of these cell cycle specific localization patterns of RFP-Killin are stable under high salt condition, consistent with Killin being tightly associated with nucleic acids within cell nuclei. Together, these cell biological results provide a molecular basis for Killin in competitively inhibiting the formation of DNA replication forks during S-phase, as well as potentially negatively regulate RNA synthesis during other cell cycle phases.

Background: Although human leukocyte antigens (HLA) have been shown in association with the outcomes of hepatitis C virus (HCV) infection among different ethnic groups, such studies remain absent in China, where the HCV prevalence is higher than the global average. Methods: In this study, 426 HCV-infected and 709 uninfected blood donors were analyzed, among whom the HLA alleles were sequenced using a high-resolution genotyping method. Results: At the 2-digit level, none of the alleles showed a statistical difference between the HCV-infected and uninfected groups. However, at the 4-digit level, the HLA-B alleles B*15:01 and B*15:02 showed an opposite association with HCV infection, i.e. B*15:01 was significantly higher in the HCV-infected group (odds ratio, OR = 1.561, p = 0.010), while B*15:02 was significantly higher in the uninfected group (OR = 0.778, p = 0.016). We also identified a higher frequency of B*13:02 in the HCV-infected group (OR = 1.515, p = 0.009) and a higher frequency of B*07:05 in the uninfected group (OR = 0.299, p = 0.001). Conclusions: The frequencies of four HLA alleles, B*07:05, B*13:02, B*15:01, and B*15:02, were found to be significantly different between the HCV-infected and uninfected blood donors in China, revealing an inverse relation of B*15:01 and B*15:02 with HCV infection. This finding suggests that the ethnic genetic variations of HLA may greatly affect the host immune responses against HCV.

To explore the molecular basis for a novel B(A) phenotype.Genomic DNA was abstracted from peripheral blood sample from the proband. ABO genotyping were carried out with sequence specific primer PCR (PCR-SSP). Exons 6 and 7 of the ABO gene were amplified with PCR and sequenced.Anti-A serum could not be adsorbed or eluted by the donor's red blood cells, and no irregular antibodies were found in the plasma. PCR-SSP showed that the ABO genotype of the donor was ABO *B/O. Sequencing results showed that one of the alleles was ABO *O02, while the other could not be defined but contained the following mutation points, 297AG, 526CG, 657CT, 701CT, 703GA, 796CA, 803GC, and 930GA. The data was accepted by the GenBank (the loading code was KM974887) and the Blood Group Antigen Mutation Database, and was confirmed to be a novel allele of B(A).A novel allele ABO *B(A)07 with 701CT has been identified, which may facilitate further study on blood antigen variants and typing of the blood groups.

Background Alloantibodies directed against antigens of the Kell blood group system are clinically significant. In the Netherlands, the KEL1 antigen is determined in all blood donors. In this study, after phenotyping of KEL:1-positive donors, genotyping analysis was conducted in KEL:1,–2 donors to identify possible KEL*02 variant alleles. Study Design and Methods A total of 407 donors with the KEL:1,–2 phenotype were genotyped for the KEL*01/02 polymorphism, followed by direct sequencing of the KEL gene if the KEL*02 allele was detected. Two K0 patients were also included. Transcript analysis was conducted in two probands with the KEL*02. M05 allele defined by a synonymous mutation (G573G). Flow cytometry analysis to determine the expression of Kell antigen was performed. Results Thirty KEL:1,–2 individuals (30/407, 7.4%) with discrepant KEL*01/02 genotype were identified. Seven novel alleles were identified: KEL*02(R86Q, R281W)mod, KEL*02(L133P)null, KEL*02(436delG)null, KEL*02(F418S)null, KEL*02(R492X)null, KEL*02(L611R)null, and KEL*02(R700X)null. Nine variant alleles described before were detected: KEL*02N.06, KEL*02N.15, KEL*02N.17, KEL*02N.19, KEL*02N.21, KEL*02M.02, KEL*02M.04, KEL*02M.05, and KEL*02(Q362K)mod. A transcript lacking Exon 16 was identified in two probands with the KEL*02M.05 allele as described before. Finally, flow cytometry analysis showed a decreased total Kell expression and a relatively increased KEL1 expression in individuals with the KEL:1,2null or KEL:1,2mod phenotype, compared to KEL:1,2 controls. Conclusion In 7.4% of a group of tested KEL:1,–2 Dutch donors, a KEL*02null or KEL*02mod allele was found. A relatively increased KEL1 antigen expression in KEL:1,2null and KEL:1,2mod individuals suggest that the expression of Kell-XK complexes depends on the availability of the XK protein.