Objective: To reveal the molecular biological mechanism of a rare Dc-variant individual using PacBio third-generation sequencing technology. Methods: ABO and Rh blood type identification, DAT, unexpected antibody screening and D antigen enhancement test were conducted by serological testing. The absorption-elution test was used to detect the e antigen. RHCE gene typing was performed by PCR-SSP, and the 1-10 exons of RHCE were sequenced by Sanger sequencing. The full-length sequences of RHCE, RHD and RHAG were detected by PacBio third-generation sequencing technology. Results: Serological findings: Blood type O, Dc-phenotype, DAT negative, unexpected antibody screening negative; enhanced D antigen expression; no detection of e antigen in the absorption-elution test. PCR-SSP genotyping indicated the presence of only the RHCE c allele. Sanger sequencing results: Exons 5-9 of RHCE were deleted, exon 1 had a heterozygous mutation at c. 48G/C, and exon 2 had five heterozygous mutations at c. 150C/T, c. 178C/A, c. 201A/G, c. 203A/G and c. 307C/T. Third-generation sequencing results: RHCE genotype was RHCE 02N. 08/RHCE-D(5-9)-CE; RHD genotype was RHD 01/RHD 01; RHAG genotype was RHAG 01/RHAG 01 (c. 808G>A and c. 861G>A). Conclusion: This Dc-individual carries the allele RHCE 02N. 08 and the novel allele RHCE-D(5-9)-CE. The findings of this study provide data support and a theoretical basis for elucidating the molecular mechanisms underlying RhCE deficiency phenotypes.

[Abstract] [Objective] To evaluate the immunogenicity of red blood cell blood group antigens in the population of Xi'an. [Methods] Data on blood group antigens of voluntary blood donors from the Shaanxi Province Blood Center and unexpected antibody detection results from clinically submitted cases between January 2019 and May 2024 were analyzed. The Giblett blood group antigen immunogenicity calculation formula was used to calculate the immunogenicity of blood group antigens based on the frequency of unexpected antibodies and the probability of antigen-negative patients receiving antigen-positive red blood cells. The relative immunogenicity of each blood group antigen was obtained by multiplying the immunogenicity of the K antigen (0.095). [Results] A total of 30 921 individuals were included for red blood cell blood group antigen analysis, with 511 cases of unexpected antibody identification. The ranking of red blood cell blood group antigen immunogenicity for the overall population was: Wr^a>E>Di^b>Fy^a>K>C>e>c>Di^a>Jk^a>M>Le^a>Jk^b>Le^b>Fy^b>S, while for males, it was: Di^b>Wr^a>E>K>Fy^a>C>e>c>M>Di^a>Jk^a>Fy^b>Le^a>Le^b>Jk^b>S. [Conclusion] Based on the immunogenicity ranking from strong to weak of red blood cell antigens in the population of Xi'an, this study provides theoretical support for the expansion and matching of antigens, and technical support for achieving precise red blood cell transfusions to improve transfusion efficacy and safety.

Objective:To establish a high-performance liquid chromatography (HPLC) characteristic spectrum of Radix Actinidiae Chinensis decoction pieces, standard decoction and formula granules; To simultaneously determine the contents of index components; To study the transfer law of formula granules based on the extraction amount, content of Catechin and characteristic spectrum of standard decoction.Methods:15 batches of Radix Actinidiae Chinensis decoction pieces were collected, the standard decoction was prepared, and the extraction amount was measured. The HPLC fingerprints of decoction pieces, standard decoction and formula granules were established, and the characteristic peaks were calibrated and attributed. The content of Catechins in decoction pieces, standard decoction and formula granules was measured, and the transfer rate law was calculated. The yield rate, the common peak transmission number of fingerprints, the content and the transfer rate were the main evaluation indexes, and the law of the transmission of the magnitude value was analyzed.Results:The paste yield of 15 batches of Radix Actinidiae Chinensis standard decoction ranged from 3.9%-6.3%. A total of 4 characteristic peaks were calibrated in the feature map, and peak 2 was identified as Protocatechuic acid and peak 4 as Catechin; 6 characteristic peaks were detected in Actinidia chinensis Planchon decoction pieces, standard decoction and formula granules, and their relative retention times were all within the specified range. The content of Catechins in Radix Actinidiae Chinensis decoction pieces was 0.4%-1.4%, which was 3.8%-4.9% in formula granules. The transfer rate of decoction pieces-standard decoction was 13.6%-38.3%, and the decoction-formula granules was 15.5%-21.2%.Conclusions:The mass transfer between Radix Actinidiae Chinensis decoction pieces, standard decoction and formula granules has a good migration. The formula granules also have a good correlation and consistency with the standard decoction, which indicating that the preparation process of Radix Actinidiae Chinensis is reasonable and feasible.

Objective:To evaluate the factors affecting the incidence of intrahepatic venovenous shunt (IVVS) in patients with cirrhosis and its impact on hepatic venous pressure gradient (HVPG).Methods:A retrospective analysis was performed on the data of patients with liver cirrhosis who received HVPG measurement in Nanjing Drum Tower Hospital from April 2013 to March 2022. Univariate and multivariate regression analyses were used to investigate the incidence rate and risk factors of IVVS and its impact on HVPG. The t-test and rank-sum test were used for the measurement data, and the χ2 test was used for the count data. Results:A total of 242 cases with cirrhosis were included in the statistical analysis, including 54 (22.3%) with IVVS and 188 (77.7%) without IVVS. There was a statistically significant difference ( P<0.05) in prothrombin time (PT), HVPG, and splenectomy history between the two groups of patients' baseline data (all P<0.05). The multiple logistic regression analysis results showed that PT was an independent risk factor for the occurrence of IVVS ( P<0.05), and patients combined with IVVS had lower HVPG values [(17.58±5.57) mmHg vs. (11.92±5.38) mmHg, 1 mmHg=0.133 kPa; t=6.623, P<0.001]. Conclusions:Patients with liver cirrhosis have a high incidence rate of IVVS, which is closely associated with a low prothrombin time. Additionally, patients combined with IVVS have low HVPG values, which affect its accuracy.

【Objective】 To evaluate the application of monoclonal typing reagents and human anti-A/B antibodies for absorption-elution test in ABO grouping. 【Methods】 The specificity of monoclonal typing reagents and human anti-A/B antibodies with standard A, B, O and AB phenotypes at 4 ℃, room temperature, and 37 ℃ were compared. Affinity was evaluated by the titer, agglutination time and agglutination intensity of the reaction with A1/B cells. 29 samples with ABO discrepancy were tested to evaluate the ability of monoclonal typing reagents and human anti-A/B antibodies to detect weak antigens in absorption-elution test. 【Results】 The specificity and affinity of human anti-A/B antibodies are low, and monoclonal typing reagents have cross reactivity. Human anti-A/B antibodies can detect most weak antigens in absorption-elution test with no cross reactivity. 【Conclusion】 In ABO grouping, the human anti A/B antibody binding absorption-elution test can serve as a supplement method for identifying ABO weak antigens. Accurate results can be obtained with reasonable reagents and corresponding methodology in serological tests,thus ensuring the safety of blood transfusion.

【Objective】 To establish a rare blood group information supply platform in Shaanxi Province. 【Methods】 The rare blood group information supply platform consists of sample registration, result registration, donor files and inventory blood. The blood donation codes of voluntary blood donors were recorded for blood typing, and the antigen identification results of each blood group system were registered, all stored in the rare blood type information supply platform. When receiving an application for unusual or rare blood type missing multiple conventional antigens or a certain high-frequency antigen, the corresponding antigen negative blood donors and their blood status (in stock or not) were queried from the donor profile module of the platform, and the inventory of blood of rare blood type was monitored dynamically. 【Results】 The results showed that 5.060% (273/5 398) of rare Rh phenotype donors, 1.540‰ (51/33 010) of donors lacking multiple regular antigens, and 13 O-type donors lacking high-frequency antigens were recorded in the rare blood type information supply platform. Among them, 0.019‰ (3/158 484) of Jk(a-b-) phenotype, 0.436‰ (2/4 586) of Di(a+b-) phenotype, and 4.030‰ (8/1 983) of Fy (a-b+) phenotype were stored in the blood bank for rare blood type. 【Conclusion】 The establishment of rare blood group information supply platform can meet the urgent demand for blood of rare blood types in clinical practice and ensure the safety of blood transfusion.

【Objective】 To observe the distribution of non-ABO-HDN and its clinical relevance, so as to provide reference for clinical diagnosis and treatment. 【Methods】 A total of 287 cases of non-ABO-HDN recorded during January 2012 to August 2022 were enrolled and tested in our laboratory. The correlation between maternal history of blood transfusion, pregnancy, unexpected antibody titers, gender, ABO-HDN and transfusion therapy was analyzed by chi-square test. 【Results】 All 287 cases of non-ABO-HDN involved 13 kinds of unexpected antibodies of 6 blood group systems. Rh-HDN accounted for 96.17% (276/287), and anti-D-HDN accounted for 47.04% (135/287). The proportion of non-ABO-HDN patients without ABO-HDN requiring exchange/transfusion was significantly higher than that of non-ABO-HDN patients with ABO-HDN(P<0.05). The ratio of need for exchange/transfusion in the high titer group (>8) was significantly higher than that in the low titer group (≤8) (P<0.05). There was no significant difference in gender, mother′s history of blood transfusion, pregnancy and whether or not to exchange/transfusion (severity of illness). 【Conclusion】 Understanding the characteristics of non-ABO-HDN and the specific distribution of unexpected antibodies, the correlation between various factors and diseases and their clinical significance are conductive to timely taking necessary intervention measures and reducing the risk of complications.

Objective To investigate the accuracy of liver stiffness (LS) as a noninvasive index in predicting hepatic venous pressure gradient (HVPG) in patients with decompensated liver cirrhosis and the value of LS in the diagnosis of decompensated liver cirrhosis. Methods A retrospective analysis was performed for the clinical data of 88 patients with decompensated cirrhosis due to viral hepatitis or decompensated alcoholic cirrhosis who received both HVPG measurement and LS measurement by acoustic radiation force impulse (ARFI) in Department of Gastroenterology, Nanjing Drum Tower Hospital, from April 2013 to June 2021, and according to HVPG, the patients were divided into serious portal hypertension (SPH) (HVPG≥20 mmHg) group with 24 patients and non-SPH (HVPG < 20 mmHg) group with 64 patients. The two groups were compared in terms of LS, spleen stiffness, portal vein velocity, and related biochemical parameters. The t -test or the Mann-Whitney U rank sum test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups. A Pearson correlation analysis was used to investigate the correlation of different noninvasive indices with HVPG, and a Logistic regression analysis was used to investigate the association of different noninvasive indices with the risk of SPH. Receiver operating characteristic (ROC) curves were plotted for different noninvasive indices in predicting HVPG≥20 mmHg, and the area under the ROC curve (AUC), sensitivity, specificity, maximum Youden index, and corresponding cut-off value were calculated to investigate the value of each index in predicting SPH. Results Among the 88 patients, 76 had decompensated cirrhosis due to viral hepatitis and 12 had decompensated alcoholic cirrhosis. There were no significant differences between the SPH group and the non-SPH group in age, sex, white blood cell count, hemoglobin, platelet count, prothrombin time, alanine aminotransferase, aspartate aminotransferase, albumin, serum sodium, creatinine, Child-Pugh class, and spleen stiffness, while there was a significant difference in LS between the two groups ( t =-3.970, P < 0.01). The correlation analysis showed that HVPG was positively correlated with LS ( r =0.458, P < 0.001). The Logistic regression analysis showed that LS was a risk factor for SPH (odds ratio=3.941, 95% confidence interval: 1.245-12.476, P =0.020). The ROC curve analysis showed that LS had an AUC of 0.751 in predicting the onset of SPH, with a sensitivity of 54.17% and a specificity of 90.63% at the optimal cut-off value of 2.295 m/s. Conclusion In patients with decompensated cirrhosis, LS measured by ARFI is correlated with HVPG and has a certain value in the non-invasive diagnosis of decompensated cirrhosis with HVPG≥20 mmHg.

Objective:To analyze the effects of two decomposition algorithms of dual-energy cone beam CT (DECBCT) (direct decomposition and iterative decomposition) on the image quality and material decomposition accuracy of different sizes of phantoms.Methods:Different sizes of imaging parts of patients were simulated using the combination of CatPhan604 phantoms and customized annuluses. CBCT with high energy of 140 kVp and low energy of 100 kVp were acquired using the Varian Edge CBCT system. Then the material decomposition of DECBCT images was performed using the two algorithms. The electron density (ED) and contrast-to-noise ratio (CNR) of each material in the CTP682 module were calculated. They were used to assess the decomposition accuracy and image quality of the two algorithms.Results:Based on the values in the Catphan604 manual, both algorithms have high ED accuracy. Only the ED accuracy of four materials of the smallest sized phantom showed statistical difference ( z = -4.21, 4.30, 2.87, 5.45, P < 0.05), but the average relative error was less than 1%. The CNR of the iterative decomposition algorithm was significantly higher than that of the direct decomposition, increasing by 51.8%-703.47%. The increase in the phantom size significantly reduced the accuracy of ED, and the increased amplitude of the relative error was up to a maximum of 2.52%. The large phantom size also reduced the image quality of iterative decomposition, and the decreased amplitude of CNR was up to a maximum of 39.71. Conclusions:Compared with the direct decomposition, the iterative decomposition algorithm can significantly reduce the image noise and improve the contrast without losing the accuracy of electron density in the DECBCT construction of different sizes of phantoms.

【Objective】 To screen individuals with rare blood type of Kidd, Diego, Duffy blood group system among the voluntary blood donor in Shaanxi province and to establish on-line and physical database of rare blood type. 【Methods】 Jk(a-b-)phenotype donors were screened by 2 mol/L urea hemolysis test. Blood donors with Di(a+ b-) phenotype were screened by genotyping; Fy(a-) and D-- phenotype donors were screened by modified antiglobulin assay. 【Results】 Three cases of Jk(a-b-) phenotype were detected out of 158 484 voluntary blood donors. The distribution frequency of Jk(a-b-) phenotype was 0.019‰. Di(a+ b-) phenotype was detected in 2(0.436‰) cases out of 4 586 voluntary blood donor. Fy(a-) phenotype was detected in 8(4.034‰) cases out of 1 983 voluntary blood donors. D-- phenotype was not detected in 29 430 voluntary blood donors. 【Conclusion】 The on-line database of Kidd, Diego, Duffy blood group system had been established by large-size screening of blood donor samples, which can conclude the region′s population distribution and genetic characteristics of RBC blood group. And physical database could further be established using the technology of red blood cells cryopreservation when the conditionspermit, so as to provide the most compatible blood for the clinical effectively improve blood transfusion safety, and provide data support for blood early warning.