Objective To investigate the distribution of A subgroups in the A and AB blood type populations among voluntary blood donors in Zhongshan,study the antigen-antibody characteristics of A subgroups,establish a local A subgroup database,and support the development of precision medicine.Methods ABO subgroup screening was performed using the microplate method.Specimens negative for monoclonal anti-A1 reactivity underwent sequencing of exons 1～7 of the ABO gene to confirm genotypes.Cryopreservation and thawing of glycerolized subgroup red blood cells(RBCs),as well as preservation efficacy of concentrated human-derived antibodies with preservatives,were studied.Results Among 1 212 blood donor specimens,28 subgroup specimens were identified,with a prevalence of 1.54%(15/971)in blood type A and 5.39%(13/241)in blood type AB.Sequencing of 10 specimens revealed 7 ABO genotypes:ABO*A2.01/O.01.02(2 cases),ABO*A1.02/B.01(3 cases),ABO*BA.02/O.01.02,ABO*AW.31.02-05/A2.05,ABO*A2.05/B.01,ABO*A2new/O.01.01,and ABO*A1.02/O.01.01(1 case each).Additionally,one rare allele mutation(c.700C>G)and one novel allele mutation(c.203G>T)(GenBank accession number:PQ152337)were identified.Human anti-A1 antibodies with a titer of 8 were successfully concentrated.Optimal preservation conditions included 0.1%preservative concentration and cryopreserved subgroup RBCs stored at 4℃for 3 days post-thaw.Conclusion The predominant A subgroups in Zhongshan donors are A2 and B(A).A preliminary database for A2 and A2B subgroups is established,along with the discovery of a novel ABO allele mutation.Cryopreservation with glycerol,PEG antibody concentration,and ProClin 300 preservative demonstrate effective applications in preserving ABO blood group antigens and antibodies.

Objective To investigate the distribution of A subgroups in the A and AB blood type populations among voluntary blood donors in Zhongshan,study the antigen-antibody characteristics of A subgroups,establish a local A subgroup database,and support the development of precision medicine.Methods ABO subgroup screening was performed using the microplate method.Specimens negative for monoclonal anti-A1 reactivity underwent sequencing of exons 1～7 of the ABO gene to confirm genotypes.Cryopreservation and thawing of glycerolized subgroup red blood cells(RBCs),as well as preservation efficacy of concentrated human-derived antibodies with preservatives,were studied.Results Among 1 212 blood donor specimens,28 subgroup specimens were identified,with a prevalence of 1.54%(15/971)in blood type A and 5.39%(13/241)in blood type AB.Sequencing of 10 specimens revealed 7 ABO genotypes:ABO*A2.01/O.01.02(2 cases),ABO*A1.02/B.01(3 cases),ABO*BA.02/O.01.02,ABO*AW.31.02-05/A2.05,ABO*A2.05/B.01,ABO*A2new/O.01.01,and ABO*A1.02/O.01.01(1 case each).Additionally,one rare allele mutation(c.700C>G)and one novel allele mutation(c.203G>T)(GenBank accession number:PQ152337)were identified.Human anti-A1 antibodies with a titer of 8 were successfully concentrated.Optimal preservation conditions included 0.1%preservative concentration and cryopreserved subgroup RBCs stored at 4℃for 3 days post-thaw.Conclusion The predominant A subgroups in Zhongshan donors are A2 and B(A).A preliminary database for A2 and A2B subgroups is established,along with the discovery of a novel ABO allele mutation.Cryopreservation with glycerol,PEG antibody concentration,and ProClin 300 preservative demonstrate effective applications in preserving ABO blood group antigens and antibodies.

【Objective】 To establish an information management platform for provincial rare blood group bank to achieve the interconnection and sharing of data such as rare blood group types, donor information and blood inventory, so as to enhance the supply security of rare blood group blood. 【Methods】 The information barriers of rare blood group management in each business link and among blood stations in Zhejiang province were analyzed using fishbone diagram. Data modeling technology was introduced based on the unified blood collection and supply business system and blood cloud platform of Zhejiang, the database view of rare blood group donors and rare blood group was constructed through demand analysis and model design, and a unified query platform was developed to achieve the sharing of rare blood group information. 【Results】 The system integrated, retrievaled and displayed rare blood type information, achieving the interconnection of information among various business links and blood stations within blood banks. Up to now, in addition to RhD negative blood group, there were 8 types of rare red blood cells in the bank, covering 7 blood group systems, 289 rare blood group donors and 216 units of frozen red blood cells of rare blood group. 【Conclusion】 The information system based on data modeling can improve the overall efficiency and collaboration of the system, and reduce the workload of system development and operation. It can provide reference for standardized data management and application, and also provide a model for establishing and improving of a national rare blood group information network.

【Objective】 To investigate the frequency of platelet antibodies in voluntary blood donors and patients in Zhongshan, Guangdong Province, and to study the specificity and cross-matching of platelet antibodies. 【Methods】 Platelet antibodies of blood donors and patients were screened by solid-phase immunoadsorption (SPIA), rechecked by flow cytometry (FCM), and antibody specificity was identified by PakPlus enzyme immunoassay, and platelet cross-matching was simulated by SPIA. 【Results】 A total of 1 049 blood donor samples and 598 patient samples were tested, with 6 (0.57%) and 49 (8.19%) samples positive for SPIA,respectively(P<0.05); In SPIA positive samples, the positive concordance rate of FCM in blood donors and patients was 100% vs 95%, and that of enzyme immunoassay was 100% vs 88%. Among the initial screening positive samples of blood donors, 5 were anti-HLA Ⅰ antibodies, accounting for 83%, and 1 was anti CD36 antibody, accounting for 17%, with an incidence rate of 0.10%. Among the 14 samples of enzyme immunoassay positive patients, 2 were anti-GP Ⅱb/Ⅲa, 1 was anti-GP Ⅱa/Ⅱa, 8 were anti HLA Ⅰ, and 3 were mixed antibodies (HLA Ⅰ, GP Ⅱb/Ⅲa, GP Ⅰa/Ⅱa). According to the types of antibodies, HLA Ⅰ antibodies were the most common, accounting for 65% (11/17), followed by HPA related anti GP, accounting for 35% (6/17). The majority of patients had a platelet antibody positive typing rate below 30%, accounting for 71.4% (10/14). 【Conclusions】 The positive rate of platelet antibody of patients in Zhongshan area is significantly higher than that of voluntary blood donors, and most of them are anti-HLA Ⅰ and anti-GP, and the incidence of anti-CD36 is extremely low. Therefore, it is necessary to establish a known platelet antigen donor bank, and at the same time, carry out platelet antibody testing and matching of patients, which is helpful to solve the issue of platelet transfusion refractoriness.

Objective To explore the clinical value of laparoscopic radical cystectomy based on fascia anatomy for bladder cancer treatment.Methods The clinical data of 51 patients with bladder cancer who underwent 3D laparoscopic radical cystectomy during Jan.2015 and Jun.2022 were retrospectively analyzed.The surgery was performed based on membrane anatomy technology along four longitudinal and two transverse planes to complete the radical cystectomy.The pelvic plexus was preserved for patients with normal preoperative sexual function.Results All surgeries were completed without conversion to open operation.The mean operation time was(502.52±108.99)min,mean intraoperative blood loss was(275.96±155.18)mL,mean postoperative drainage time was(4.14±2.41)d,and the mean postoperative hospital stay was(16.37±4.85)d.The mean number of lymph nodes removed was(17.98±11.48).The mean postoperative follow-up was(30.27±19.39)months.At the last follow-up,no Clavien ≥grade 3 complications were observed.The estimated overall survival(OS),tumor-specific survival(TSS),and recurrence-free survival(RFS)were 82.4％,92.2％,and 88.2％,respectively.The lymph node positive patients had shorter OS and RFS(60.0％,60.0％)than the lymph node negative patients(84.8％,91.3％).Among the 19 male patients who underwent radical cystectomy with pre-exposure and preservation of pelvic plexus,daytime and nocturnal continence rate were 83.3％and 72.2％,respectively,and 17 patients recovered potency within 6 months postoperatively.Conclusion Laparoscopic radical cystectomy based on fascia anatomy is safe and effective in laparoscopic radical cystectomy,with standardized surgical procedure,satisfactory oncological outcomes,little hemorrhage,few complications and fast recovery.

OBJECTIVE: To analyze the serological characteristics and molecular mechanism for an individual with p phenotype. METHODS: An individual with p phenotype upon blood group identification at Jiaxing Blood Center in May 2021 was analyzed. ABO, RhD and P1PK blood groups and irregular antibodies in her serum were identified using conventional serological methods. The encoding region of α1, 4-galactosyltransferase gene (A4GALT) encoding P1 and Pk antigens was analyzed by polymerase chain reaction-sequence-based typing (PCR-SBT). RESULTS: The individual was A group, RhD positive and had a p phenotype of the P1PK blood group system. Anti-PP1Pk was discovered in her serum. Sequencing analysis revealed that she has harbored a homozygous c.343A>T variant of the A4GALT gene. CONCLUSION The homozygous c.343A>T variant of the A4GALT gene probably underlay the p phenotype in this individual.
Female ; Animals ; Blood Group Antigens ; Homozygote ; Phenotype ; Polymerase Chain Reaction ; Sequence Analysis, DNA

OBJECTIVE: To explore the genetic mechanism underlying a case with para-Bombay phenotype. METHODS: The ABO and Lewis phenotype were identified with serological methods. The coding regions of exons 6 and 7 of the ABO and FUT1 genes were amplified with PCR and directly sequenced. Haploid sequence analysis was carried out on the variant sites of the FUT1 gene. RESULTS: Serological analysis confirmed that the proband has a rare para-Bombay phenotype. Direct sequencing revealed that he was a B.01/O.01.02 heterozygote for the ABO gene, and had heterozygous deletion for the 768 and 881-882 sites of the FUT1 gene. Further haploid analysis showed that the c.881_882delTT deletion has occurred in one haploid while c.768delC was present in the other haploid. The proband was therefore determined as a FUT1*01N.13/01N.20 heterozygote, which have resulted in frameshift in polypeptide chain p.Phe294Cysfs*40 and p.Val257Phefs*23, respectively. CONCLUSION A rare bi-allelic heterozygous deletion of para-Bombay phenotype has been identified in a blood donor. The c.881_882delTT and c.768delC deletions may decrease the activity of α-1,2-fucosyltransferase.
Animals ; Male ; ABO Blood-Group System/genetics* ; Alleles ; Fucosyltransferases/genetics* ; Genotype ; Heterozygote ; Mutation ; Phenotype ; Humans

Objective:To investigate the effect of virus inactivation on weak positive result of 2019 novel coronavirus(2019-nCoV) nucleic acid test.Methods:A retrospective study was conducted on the nasopharyngeal swabs of three patients with positive PCR nucleic acid test for 2019-nCoV at different concentrations in the Second affiliated Hospital of Zhejiang University Medical College from January to February 2020.The virus in nasopharyngeal swab specimens were inactivated by water bath at 56 ℃ for 30 min, dry bath at 56 ℃ for 60 min and dry bath at 60 ℃ for 30 min respectively. After treatment, these samples RNA were extracted and then detected by three new commercial quantitative real-time polymerase chain reaction reagent kits for 2019-nCoV.Cycle threshold (Ct) value was used to evaluate the effect of virus inactivation on nucleic acid detection of 2019-nCoV.Results:There was no significant difference between the groups before and after inactivation. Ct values of ORF1ab gene before inactivation were 23.28±0.28, 25.25±0.25, 28.93±0.44, 32.06±0.47, 35.20±0.38, 32.89±0.38, 36.24±0.23, 33.30±0.46, and those after inactivation were, group 1:23.60±0.20, 27.29±0.30, 31.83±0.51, 37.41±0.46, group 2: 24.25±0.34, 27.18±0.42, 31.84±0.61, 34.99±1.01, 34.89±0.45,group 3: 23.37±0.17, 26.89±0.52, 32.05±0.50.Ct value of N gene before inactivation were 24.38±0.09, 26.64±0.11, 30.35±0.12, 33.29±0.33, 36.93±0.11, 34.50±0.12, 35.63±0.12, those after inactivation were, group 1: 24.66±0.11, 28.52±0.14, 32.71±0.14, 37.00±0.13;group 2: 25.41±0.10, 28.79±0.15, 33.29±0.28; group 3: 23.37±0.11, 28.68±0.11, 33.54±0.13, 37.18±0.23(ORF1ab gene: t=-1.416; N gene: t=-1.379, P>0.05). There was no significant difference among the three inactivation groups, the specific Ct values are shown above(ORF1ab gene: t=-0.460; N gene: t=-0.132, P>0.05). However, the Ct values of the inactivated groups (1,2,3) and the non-inactivated group at different dilution times were different (10 ×:Ct value of ORF1ab was 25.25±0.25 in the non-inactivated group, and 27.29±0.30, 27.18±0.42 and 26.89±0.52 in the inactivated group1,2 and 3, t(ORF1ab)=-7.327, P<0.01.Ct value of N gene in the non-inactivated group was26.64±0.11, those in inactivated group 1, 2 and 3 were 28.52±0.14, 28.79±0.15 and 28.68±0.11, respectively, t (N)=-19.340, P<0.01. 100 ×:Ct value of ORF1ab was 28.93±0.44 in the non-inactivated group, and 31.83±0.51,31.84±0.61 and 32.05±0.50 in the inactivated group1,2 and 3, t (ORF1ab)=-9.462, P<0.01. Ct value of N gene in the non-inactivated group was 30.35±0.12, those in the inactivated group 1, 2 and 3 were 32.71±0.14, 33.29±0.28 and 33.54±0.13, respectively, t (N)=-18.583, P<0.01. The positive detection rate of the non-inactivated group (7/11, 8/11, 5/11) was significantly different from that of the inactivated group (inactivated group 1:4/11, 4/11, 3/11, inactivated group 2:3/11, 3/11, 3/11, and inactivated group 3:3/11, 3/11, 2/11) ( Z=-2.670, P<0.01). There were no significant difference among the inactivated groups(inactivated group 1:4/11, 4/11, 3/11, inactivated group 2:3/11, 3/11, 3/11, inactivated group 3:3/11, 3/11, 2/11) ( Z=4.413, P>0.05) and among the three reagents(reagent 1:7/11, 4/11, 3/11, 3/11, reagent 2:8/11, 4/11, 3/11, 3/11, reagent 3:5/11, 3/11, 3/11, 2/11)(χ 2=1.199, P>0.05). Conclusion:The virus inactivation can degrade the nucleic acid of the 2019-nCoV, resulting in the decrease of the Ct value and the false negative results of the low-concentration specimens.

Human infection with avian influenza A (H7N9) is an acute contagious respiratory disease. Acute respiratory distress syndrome (ARDS) is a common complication in patients with severe avian influenza A (H7N9), for whom mechanical ventilation (MV) is an important supportive method. A patient, suffered from severe avian influenza A (H7N9) and complicated with ARDS, was admitted to the Second Affiliated Hospital of Guizhou Medical University in January 2017. With very intensive care for oxygenation, respiration and consciousness, and monitoring, she was successfully cured by comprehensive managements, among which noninvasive mechanical ventilation (NIV) was the major respiratory support method. The result demonstrate that, in patients with conscious state, satisfied expectoration ability and relatively good cooperation, and with close observation of oxygenation and respiratory rate, NIV may be accepted as an effective method for patient with ARDS caused by severe avian influenza A (H7N9).

Objective To compare the short‐term clinical effects of single segment and two segments of Dynesys dynamic in‐ternal fixation system for treating lumbar degenerative disease .Methods 40 cases of degenerative lumbar disease treated by the Dynesys dynamic fixation system combined with posterior approach decompression from 2009 July to March 2012 were selected and performed the evaluation on the clinical effects by the waist and leg pain visual analogue scale (VAS) ,the Oswestry disability index (ODI) and the orthopaedic spinal association of North America (NASS) index .The imaging method was used to observe the mobili‐ty of operative segment and proximal adjacent segment and the degenerative change of intervertebral height .Results All of the 40 cases were followed up .The VAS score and ODI score at postoperative 3 months and last follow‐up were significantly decreased when compared with the pre‐operative scores (P<0 .05);there was no statistically significant differences in the VAS score and ODI score at 3 months and last follow‐up between the single segment group and the 2‐segment group (P> 0 .05) .The NASS index showed no statistically significant difference in the postoperative effect satisfaction between the two groups (P>0 .05);there was no statistically significant difference in the variation of the intervertebral height between before operation and at the last follow‐up (P>0 .05) .There was statistically significant difference in the motility of proximal adjacent segment at the last follow‐up between the two groups (P<0 .05);but comparing the motility of proximal adjacent segment between the last follow‐up and pre‐operation , only the single segment group showed statistically significant difference (P< 0 .05) .Conclusion The Dynesys dynamic fixation system combined with posterior approach decompression operation has good clinical effect in the treatment of lumbar degenerative disease .Meanwhile ,the single segment method has more significant effect on the motility of proximal adjacent segment than the double segments method .