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Objective To investigate the importance of a nomogram model based on biomarkers and CT signs in the prediction of the invasive risk of ground glass nodules. Methods A total of 322 patients with ground glass nodule, including 240 and 82 patients in the model and verification groups, respectively, were retrospectively analyzed. Independent risk factors for the invasive risk of ground glass nodules were screened out after using single and multiple Logistic analysis. R software was used to construct the nomogram model, and clinical decision curve analysis (DCA), receiver operating curve (ROC), and calibration curve were used for internal and external verification of the model. Results In this study, the independent risk factors for the invasive risk of ground glass nodules included systemic immune-inflammation index (SII), CYFRA21-1, edge, vascular cluster sign, and nodular consolidation tumor ratio (CTR). The area under the ROC curve of the constructed nomogram model had a value of 0.946, and that of the external validation group reached 0.932, which suggests the good capability of the model in predicting the invasive risk of ground glass nodules. The model was internally verified through drawing of calibration curves of Bootstrap 1000 automatic sampling. The results showed that the consistency index between the model and actual curves reached 0.955, with a small absolute error and good fit. The DCA curve revealed a good clinical practicability. In addition, nodule margin, vascular cluster sign, and CTR were correlated with the grade of pathological subtype of invasive adenocarcinoma. Conclusion A nomogram model based on biomarkers and CT signs has good value and clinical practicability in the prediction of the invasive risk of ground glass nodules.
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Purpose To investigate the clinicopathological features,immunophenotype,molecular changes,differential di-agnosis,treatment and prognosis of the micropapillary subtype of serous borderline tumor(MSBT)in the ovary.Methods The clinical and pathological data of 14 cases of ovarian MSBT.Im-munohistochemical EnVision staining was used to detect the ex-pression of IMP3.BRAF and KRAS mutations were detected by qRT-PCR and Sanger sequencing,respectively.Its clinical and pathological characteristics were analyzed with review of relevant literature.Results The age of the patients ranged from 27 to 56 years,with mean 41.7 years.Nine cases had bilateral ovari-an masses.Preoperative serum CA125 increased in 11 cases.On gross examination,the cut section was cystic and solid with intracystal papillae.Microscopically,all cases showed a papilla-ry structure,with the characteristic elongated micropapillae radi-ating directly from the cyst wall or large unbranched papillae.The length to width ratio of the papillae was greater than 5.The cells covering the papillae were cubic to polygonal.Mild to mod-erate atypia was noted with a range of＞5 mm in the micropapil-lary area.Five cases had microinvasion.Six cases had non-in-vasive peritoneal seeding.Five cases were accompanied by asci-tes,and atypical tumor cells were observed in ascites.Three ca-ses had lymph node involvement.Nine cases had psammoma bodies.Immunohistochemically,the tumor cells were positive for ER,PR,CA125,CK7 and WT-1;p53 was wild type,HER2 and IMP3 were negative,and Ki67 was positive in 5％to 30％.KRAS mutations were detected in 3 of 14 cases,inclu-ding G12C,G12D and Q70(nonsense mutation).No BRAF V600E mutation was detected,and 1 case had BRAF T559I mu-tation.Seven patients underwent radical surgery and 7 patients underwent conservative surgery without special treatment after surgery.Five patients had a history of recurrence,and the fol-low-up time ranged from 1 to 12 years.Conclusion MSBT has special morphology,often bilateral,and is prone to peritoneal implantation and recurrence.It should be distinguished from classical ovarian serous borderline tumor.
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BACKGROUND:It has been hypothesized that PANoptosis may be involved in the pathologic process of osteoporosis,but there have been no studies addressing the mechanisms of PANoptosis genes in osteoporosis. OBJECTIVE:To analyze the biological mechanism of PANoptosis regulators in the occurrence and development of osteoporosis. METHODS:The GSE56815 dataset was obtained from the Gene Expression Omnibus database and PANoptosis genes were extracted for differential analysis.The key genes of PANoptosis were screened by random forest tree model to construct a disease risk prediction model.Consensus clustering algorithm,single sample genome enrichment analysis and immune infiltration analysis were used to explore the differences between different PANoptosis molecular subtypes.Herbal drugs that regulate the key genes of PANoptosis were predicted through Coremine medical database,a medical ontology information retrieval platform. RESULTS AND CONCLUSION:Based on the four PANoptosis key genes(CASP1,CASP10,MEFV,and TNF),the diagnostic markers of osteoporosis were determined,and the risk prediction model was constructed and verified.Osteoporosis was divided into two different PANoptosis subtypes(clusters A,B and gene clusters A,B),and the PANoptosis scores of cluster B and gene cluster B were higher than those of cluster A and gene cluster A,respectively.Traditional Chinese drugs such as ginseng which can regulate the key genes of PANoptosis were predicted by the Coremine medical database.
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BACKGROUND:Increasing studies have found that estrogen has a certain correlation with tendinopathy,but for a long time,there are few experiments and summaries of estrogen in tendinopathy,which makes it difficult for specialists and scholars in related fields to fully understand the research status. OBJECTIVE:To summarize the current clinical or preclinical original research,so as to summarize the role of estrogen in tendinosis,and make a certain prospect for the evaluation and management of estrogen in tendinosis in the future. METHODS:Relevant literature in PubMed,Web of Science,CNKI,WanFang,and VIP databases were searched by computer.Search time was from January 2008 to September 2023.The search terms were"oestrogen,estrogen,estrogen receptor,tendinopathy,tendonopathy,sinew,tendon,tendons,myotenositis"in English and"estrogen,estrogen receptor,tendinosis,tendon,tendinitis"in Chinese.According to the selection criteria,the search results were screened and excluded,and finally 60 documents were included for review and analysis. RESULTS AND CONCLUSION:In vivo studies have shown that estrogen can promote tendon anabolism.In vitro experiments have also proved that various estrogens can promote the proliferation of tendon cells and reduce inflammation and apoptosis,but most of the experiments are limited to animal models.Estrogen receptor β acts more in tendon injury and repair processes,but estrogen receptor α has not been found to have a major impact on tendon injury.The expression of estrogen receptor β can repair the tendon by affecting the formation of fat,the deposition of type I collagen and reducing the apoptosis of tendon cells,while its over-expression may promote inflammation and angiogenesis,thus promoting the inflammatory process and playing a role in tendon injury.Animal studies have shown that estrogen deficiency may reduce the synthesis efficiency of collagen in the tendon,decrease the elasticity of tendon,inhibit the synthesis and metabolism of the tendon,which is not conducive to the repair of tendon injury,while normal level of estrogen may stimulate the synthesis of type I collagen in tendon and promote the proliferation and metabolism of tendon cells.At present,the molecular mechanism of estrogen in tendon injury has not been fully explained.More experiments focus on tendon collagen synthesis,cell proliferation and apoptosis.Only a few documents have studied the molecular mechanisms of estrogen receptor β deficiency regulating interferon regulatory factor 5-chemokine ligand 3 axis,E2 regulating estrogen receptor α and PI-3K-Akt signaling pathways,and high levels of estradiol reducing the level of free-circulating insulin-like growth factor.Various estrogens,including endogenous estrogens and phytoestrogens,are beneficial to the repair of tendinopathy at normal levels,and estrogen receptor β mainly affects the formation of fat,the deposition of type I collagen and the reduction of apoptosis of tendon cells through,which lays a foundation for the future treatment of tendinopathy with different subtypes of estrogens in vivo and the influence of estrogen membrane receptors on tendinopathy.
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Purpose To compare the CT imaging features of the novel coronavirus Omicron variant and influenza A-H1N1-associated viral pneumonia,and to investigate the factors associated with the uptake process of the two pneumonias.Materials and Methods A total of 43 patients with Omicron virus pneumonia(Omicron group)and 30 patients with influenza A(H1N1)virus pneumonia[influenza A(H1N1)group]in Civil Aviation General Hospital from December 2022 to March 2023 were retrospectively collected.The clinical data of the two groups were compared,including age,gender,symptoms(fever or not),duration of symptoms and incidence of complications.White blood cells,monocytes,lymphocytes,neutrophils,C-reactive protein,etc.]and initial and follow-up CT imaging features[lesion density,distribution,signs and qualitative CT severity score(CTSS)].Results The mean age of patients in Omicron group was higher versus that in H1N1 group[(68.61±15.94)years vs.(51.20±16.39)years,P<0.000 1],and the fever rate in Omicron group(58.1%vs.86.7%,P=0.009)and monocyte count[(0.40±0.16)vs.(0.58±0.19),P<0.000 1]were lower than those in the influenza A(H1N1)group.Chest CT showed that the lesions of patients in the Omicron group were mainly distributed under the pleura,and the lesions of patients in the influenza A(H1N1)group were mainly distributed under the pleura and along the bronchovascular bundle(χ2=8.592,P=0.035).Patients in the Omicron group were more likely to have interlobular septal thickening(χ2=11.753,P=0.001),paving pattern(χ2=16.216,P<0.000 1),air bronchogram(χ2=16.216,P<0.000 1),pleural effusion(P=0.039)and pleural thickening(χ2=4.067,P=0.044)than patients in the influenza A(H1N1)group,while patients in the influenza A(H1N1)group were more likely to have nodules than those in the omicron group(χ2=6.971,P=0.008).The CTSS scores of patients in the omicron group were higher than those in the influenza A(H1N1)group at the initial diagnosis(Z=413,P=0.009)and follow-up(Z=107,P=0.027).The correlation between the change of follow-up CTSS and the initial CTSS in the Omicron group was the strongest(r=0.689,P<0.000 1).There was the strongest correlation between the change of follow-up CTSS and the duration of symptoms in influenza A(H1N1)group(r=0.954,P<0.000 1).Conclusion Patients in the Omicron group have a higher range of initial and follow-up lesions than those in influenza A(H1N1)group,and the degree of pneumonia absorption in the omicron group may be related to the initial CTSS,whereas in the influenza A(H1N1)group it may be related to the duration of symptoms.
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Objective To investigate the effect of electroacupuncture at Lianquan point on nerve function deficit in post-stroke dysphagia(PSD)rats and its potential effect on regulating of transi-ent receptor potential vanilic acid subtype 1(TRPV1)signaling pathway.Methods A total of 60 male SD rats with SPF grade were randomly divided into a normal group of 12(only mild inser-tion of the thread,which did not lead to intracerebral artery occlusion),and the remaining 48 rats were established into PSD models.The 36 rats successfully made were randomly divided into mod-el group,treatment group and treatment+caffeic acid group,with 12 rats in each group.The la-tency and frequency of swallowing were recorded.Biological signal collector was applied to detect hypoglossal nerve discharge,lingual muscle threshold intensity,and contraction amplitude.ELISA was employed to detect the content of substance P in serum,toluidine blue staining was conducted to count the number of Nissl bodies in the hypoglossal nucleus,and immunohistochemistry was applied to measure the expression levels of TRPV1,serotonin(5-HT),phosphorylation(p)-p38,and neuronal nitric oxide synthase(nNOS)in the hypoglossal nucleus.Results Compared with the normal group,the model group had shorter swallowing latency,less swallowing frequency,and decreased integrated area of hypoglossal nerve discharge,amplitude of tongue muscle contrac-tion,serum substance P content,threshold strength of tongue muscle,number of Nissl bodies,and the expression levels of TRPV1 and 5-HT,but increased threshold intensity of tongue muscles and expression levels of p-p38 and nNOS in the hypoglossal nucleus(P＜0.05).Compared with the model group,the amplitude of tongue muscle contraction,serum substance P content,number of Nissl bodies,and the expression levels of TRPV1 and 5-HT proteins in rats in the treatment group were increased[2.36±0.26 vs 1.77±0.22,3.46±0.36 vs 2.15±0.18,(3.92±0.38)ng/ml vs(1.69±0.17)ng/ml,(33.60±3.65)vs(24.60±2.34),(19.85±2.11)％vs(9.79±1.07)％,(22.43± 2.34)％vs(10.85±1.13)％,P＜0.05].Conclusion Electroacupuncture at Lianquan point may improve neurological deficits in PSD rats by activating the TRPV1 signaling pathway.
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Mild cognitive impairment(MCI)is a prodromal phase of dementia with heterogeneity in etiology, clinical presentation, disease progression, outcome, and prognosis.The number of studies on MCI subtypes is increasing each year.This article discussed the subtypes of MCI from the perspectives of phenotypic characteristics, etiology, progression, outcome, and data-driven approaches, and further summarizes the epidemiological characteristics, influencing factors, and risk of progression to dementia of each subtype.Despite the increasing number of studies on MCI subtyping, research remains limited on the correlation between MCI subtypes from different perspectives, indicating a need for further investigation in order to achieve more accurate and effective diagnosis and treatment of MCI and obtain evidence for dementia prevention.
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The incidence of lung micropapillary adenocarcinoma is low, and it is easy to relapse and metastasize after operation, which leads to poor prognosis. This article mainly summarizes the imaging and histopathological features of lung micropapillary adenocarcinoma and discusses the effects of surgical procedures, chemotherapy and targeted drug therapy on the prognosis of patients, in order to deepen the understanding of lung micropapillary adenocarcinoma.
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Objective@#To investigate the molecular transmission network characteristics of HIV-1 among men who have sex with men (MSM) in Shaoxing City, Zhejiang Province, so as to provide insights into AIDS prevention and control among MSM.@*Methods@#Newly reported HIV/AIDS cases among MSM in Shaoxing City were selected from January 2021 to September 2023. Plasma samples before the antiviral treatment were collected. The HIV-1 pol gene was amplified using reverse transcription PCR and nested PCR to construct phylogenetic trees for gene subtype analysis. The HIV-TRACE method was used to construct a molecular transmission network with a genetic distance of 1.5% to analyze clustering and the characteristics of cases within molecular clusters.@*Results@#A total of 216 HIV/AIDS cases among MSM were included, and 179 qualified sequences were obtained. The predominant HIV-1 subtypes were CRF07_BC and CRF01_AE, with 95 and 66 cases, respectively. At 1.5% genetic distance, 20 molecular clusters were formed, with 61 nodes and 58 edges. A total of 61 sequences were connected to the transmission network (34.08%). HIV/AIDS cases among MSM from all the counties (cities, districts) in Shaoxing City were included in the network. There was the largest molecular cluster with ≥10 nodes, involving 12 cases from five counties (cities, districts), 3 medium-sized molecular clusters with 4 to 5 nodes, and the 16 small-sized molecular clusters with 2 or 3 nodes. Seven cases with high risk of transmission, each with ≥4 edges, were all CRF07_BC subtypes. Among them, two cases were from the large molecular cluster, and five cases were from the same molecular cluster composed of cases from Shengzhou City and Xinchang County.@*Conclusions@#The predominant HIV-1 subtypes among MSM in Shaoxing City were CRF07_BC and CRF01_AE. There was cross-regional HIV transmission, and potential transmission risk might exist in Shengzhou City and Xinchang County.
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【Objective】 To correctly identify the blood group of ABO and study its molecular biological characteristics. 【Methods】 Blood samples from blood donors with discrepancies in forward and reverse typing using the microplate method were subjected to both saline tube agglutination test for serological identification and polymerase chain reaction-sequence specific primers (PCR-SSP) for genotyping. Additionally, direct sequencing of exons 1 to 7 of the ABO gene was performed on some donor samples to analyze their blood phenotype, genotype and gene sequence. 【Results】 For 256 samples showing discrepancy between forward and reverse typing in microwell method, 119 were identified as normal ABO blood types, 90 were weakened ABO antibodies and 47 were ABO subtypes by serology tube test. According to the PCR-SSP genotyping test, 233 of 256 (91.02%) were consistent with serological phenotype and genotype, 17 of 256 (6.64%) were inconsistent, and 6 samples can′t read genotype based on the kit result typing table.A total of 17 genotypes were identified in 250 samples as AO1 in 56, AO2 in 58, AA in 50, BO1 in 31, BO2 in 17, BB in 8, O1O1 in 2, O1O2 in 7, AB in 13, AO4 in 1, A205O2 in 1, A205A in 1, A201A in 1, O1O4 in 1, O2O2 in 1, A201B in 1 and A205B in 1. Sequencing of exons 1 to 7 of the ABO gene was carried out in 78 samples, and 29 ABO alleles were detected, seven of which were common alleles (*A101, *A102, *A104, *B101, *O01, *O02, *O04), 22 of which were rare alleles (*A201, *A205, *Ax01, *Ax03, *Ax13, *Ax19, *Ax22, *Ael10, *B305, *Bel03, *Bel06/*Bx02, *Bw07, *Bw12, *Bw17, *Bw37, *O05, *O26, *O61, *B(A)04, *B(A)07, *cisAB01 and *cisAB01var). In addition, six rare allele mutation sites were identified (c.101A>G; c. 103_106delG; c. 146_147insGC; c. 259G>T; c. 322C>T; c. 932T>C). 【Conclusion】 The identification of ambiguous ABO blood group requires the combination of serological testing and molecular biological examination to correctly identify the blood type and ensure the safety of clinical blood transfusion.
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【Objective】 To investigate the serological and molecular biology characteristics of ambiguous blood types among voluntary blood donors in Putian, and to file records for relevant donors for future reference in case of their own transfusion needs or emergency donations to others. 【Methods】 A total of 68 593 blood samples from voluntary blood donors in Putian Central Blood Station between January 1, 2019 and August 31, 2023 were collected and tested for blood types using serological methods. ABO gene (including promoters, enhancers and seven exons as well as their flanking sequences and intron 6) and FUT1 gene testing were performed on ambiguous blood types.3D models were constructed using Chimira and PyMOL software to predict the impact of gene mutations on enzyme structure. 【Results】 A total of 16 ABO subtypes were identified by serological methods, with the highest detection rate as the para-Bombay phenotype (0.73 per 10 000), followed by the cisAB phenotype (0.44 per 10 000). Gene analysis revealed 12 cases with known mutations (4 cases of FUT1.01N.06/FUT1.01N.06, 1 case of FUT1 01W.08/FUT1.01N.06, 2 cases of A2.08/B.01, 1 case of BA.02/O.01.01, 1 case of A3.07/O.01.01, 3 cases of cisAB.01/B.01), and key mutations were not found in 4 cases (2 cases of A1.02/B.01, 2 cases of A1.02/O.01.02). 3D molecular model analysis revealed that both A3.07 and FUT1.01W.08 allele can lead to a decrease in activity of the corresponding glycosyltransferases, resulting in the emergence of subtypes. 【Conclusion】 The most common phenotype causing discrepancies in ABO blood type testing among voluntary blood donors from Putian is the para-Bombay phenotype, with the most common allele being FUT1.01N.06.
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【Objective】 To investigate the serological changes of blood type and whether hemolytic reactions occurred in a patient with ABw subtype after type A therapeutic plasma exchange(TPE) retrospectively. 【Methods】 The ABO blood group identification and genotyping was carried out by microcolumn gel method, microcolumn glass bead method, saline tube method and absorption-elution test, and the laboratory results before and after TPE were analyzed. 【Results】 The patient underwent 2 000 mL type A TPE in other hospitals and was transferred to our hospital. On the second day of hospitalization, the test showed consistency between forward and reverse blood typing. On the third day, the detection showed that Bc was ±~1+ , and discrepancy was found between forward and reverse typing. The presence of B antigen was confirmed through tube method and human anti-B absorption-elution test, and was identified as ABw03 by molecular biological method. After type A TPE, Hb and Plt both temporarily decreased, creatinine slightly increased, ALT and AST continued to decrease, while TBIL, DBIL and IBIL showed a temporary decrease followed by an increase, indicating a slight hemolytic reaction. 【Conclusion】 Infusion of large volume of plasma may cause hemolytic reactions when ABO subtype patients contain weak A or B antigens.The blood type should be strictly identified according to the ABO blood type forward and reverse typing standards. Additional laboratory tests combined with molecular biology techniques are required for weak agglutination, which can reduce the missed detection of ABO subtypes and reduce the occurrence of transfusion reactions.
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【Objective】 To study the changes in serum immunoglobulin levels in children with thalassemia who undergo repeated blood transfusions and explore their correlation with delayed hemolytic transfusion reactions(DHTR). 【Methods】 Serum samples from children with thalassemia who received blood transfusion treatment from June 2022 to April 2023 (observation group) and healthy children who underwent physical examination (control group) in our hospital were collected. The levels of serum immunoglobulins (IgG subtype, IgM, IgA, IgE and IgD) were detected using flow cytometry CBA multi-factor quantitative detection technology, and the differences between the two groups were compared. The children were divided into 4 groups according to different transfusion numbers: ≤10 numbers, 11-30 numbers, 31-50 numbers and >50 numbers, and the differences between different blood transfusion numbers and serum immunoglobulin levels in each group were compared using one-way analysis of variance (ANOVA). Children with thalassemia with DHTR were in the hemolysis group, and children with thalassemia who did not experience DHTR were in the non-hemolysis group. The changes in serum immunoglobulins (IgG subtypes, IgM, IgA, IgE and IgD) between the two groups were compared to explore the correlation between serum immunoglobulins in thalassemia children with repeated transfusion and DHTR. 【Results】 The levels of IgG1, IgG3, IgG4 and IgA in the observation group were significantly higher than those in the control group, with the increase of(2.07±2.12), (0.67±2.03), (0.30±0.37)and(6.04±11.40)mg/mL, respectively, while the level of IgD in observation group was significantly lower than that in the control group, with a decrease of(0.03±0.01)mg/mL, P0.05). IgG1 and IgG4 both significantly increased with the number of blood transfusions.The IgG1 in the 4 groups increased sequentially as(0.30±0.62), (0.41±0.51)and(3.60±3.48)mg/mL, and IgG4 increased sequentially as (0.12±0.13), (0.22±0.07) and (0.21±0.38)mg/mL. IgG2, IgM and IgD showed a significant decrease, with IgG 2, IgM, and IgD in four groups decreased as(0.91±1.50), (0.14±0.10)and(0.05±0.05)mg/mL, respectively, showing significant differences with the number of blood transfusions(P0.05). IgG1, IgG3 and IgG4 in the hemolysis group were significantly higher than those in the non-hemolysis group, with an increase of (4.44±3.41), (0.73±1.26)and(0.52±0.40), respectively(P0.05). 【Conclusion】 The serum immunoglobulin levels of children with thalassemia who undergo repeated blood transfusions are abnormal. There are differences in correlation between the number of blood transfusions and serum immunoglobulin levels among children with thalassemia who undergo repeated blood transfusions. The relevant serum immunoglobulins for DHTR in children with thalassemia who undergo repeated blood transfusions are IgG1, IgG3 and IgG4.
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【Objective】 To study the relationship between ABO subtype, para-Bombay blood group and genotype, so as to explore the possible molecular mechanism of these two blood groups, and provide accurate genetic detection targets and theoretical basis for the accurate identification of ABO blood group. 【Methods】 First, the serology of 24 200 patients with blood type identification in the Ruijin Hospital from February to December in 2022 were analyzed, as well as 10 ambiguous ABO samples from other hospitals(3 were suspected ABO subtype and 7 were suspected para-Bombay blood group). Then ABO subtypes and para-Bombay blood groups were directly sequenced or post-clonal sequencing was performed to analyze ABO, FUT1 and FUT2 gene sequences. 【Results】 Among the 24 200 patients underwent blood type identification, 7 cases of ABO subtypes were detected. Among the 10 ambiguous samples sent by other hospitals, 2 of ABO subtypes, 1 of normal type A, and 7 of para-Bombay blood type were detected. In total, we identified blood types as follows: 1) 9 ABO subtypes: Ael(AEL.02/O.01.02), AelB(AEL.05/B.01), three of B3(2 of B3.03/O.01.01, 1 of B3.03/O.01.02), B(A)(BA.02/O.01.01), ABweak(A1.02/BW.07), Bweak(BW.31 /O.01.02), A2Bweak(A2.05 /BW.31); 2) 7 para-Bombay blood group: ABmh (FUT1*01N.13/FUT1*01N.13), 4 of Amh (3 of FUT1*01N.06/FUT1*01N.13, 1 of FUT1*01N.13/FUT1*01N.13); two of Bmh (FUT1*01N.06 /FUT1*01N.06, FUT1*01N.06/FUT1*01N.13), all of FUT2 of the 7 cases were FUT2*01/FUT2*01. 【Conclusion】 Clinical ABO blood group variant samples need to be identified in combination with serological and molecular biology to improve the accuracy of identification, thus providing a reference for safe blood transfusion, organ transplantation, and the prediction and prevention of fetal-maternal immune hemolytic disease.
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【Objective】 To study the molecular mechanism of 95 samples of serological ABO subtypes. 【Methods】 A total of 95 samples with discrepancy between forward and reverse blood grouping were subjected to serological confirmation, and genotyped by polymerase chain reaction with sequence-specific primers (PCR-SSP). For those subtype alleles could not be detected by PCR-SSP, ABO gene exon 1-7 sequencing and gene single strand sequencing were performed successively to determine the mutation site and the gene location. 【Results】 A total of 34 ABO alleles were detected in 95 samples. Five common ABO alleles (ABO*A1.01, ABO*A1.02, ABO*B.01, ABO*O.01.01 and ABO*O.01.02) and 29 rare ABO alleles were identified, including 16 named alleles by ISBT (ABO*A2.01, ABO*A2.05, ABO*A2.13, ABO*A3.07, ABO*AW.37, ABO*AEL.05, ABO*B3.01, ABO*B3.05, ABO*BW.03, ABO*BW.07, ABO*BW.27, ABO*BEL.03, ABO*cisAB.01, ABO*cisAB.05, ABO*BA.02, ABO*BA.04) and 5 named alleles by dbRBC(A223, B309, Bw37, Bel09, Bw40)and eight unnamed alleles [ABO*B.01+ 978C>A, ABO*A1.02+ 248A>T, ABO*B.01+ 125dupT, ABO*B.01+ (98+ 1G>A), ABO*A1.02/ABO*B.01+ 1A>G, ABO*A1.02/ABO*O.01.01+ 28G>T, ABO*A1.02/ABO*B.01+ 538C>T, ABO*A1.02/ABO*O.01.01+ 797insT] .The last four samples could not be verified by single strand because of insufficient samples. In 95 samples, 76 samples (21 named alleles of ISBT and dbRBC) were identified by PCR-SSP, and the remaining 19 samples were identified by exon 1-7 sequencing of ABO gene, of which 8 were identified as unnamed alleles, and the remaining 11 samples were not identified as subtype alleles. 【Conclusion】 The molecular genetic mechanism of 95 serological ABO subtypes was revealed, and 8 rare novel alleles were identified. The detection of ambiguous blood groups is influenced by factors such as patient pathology and physiology, therefore the combination of serological testing and genetic testing is suggested for the identification of ABO subtype.
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【Objective】 To identify and analyze a case of ABO discrepancy between forward and reverse blood grouping, and to provide reference for the identification of ambiguous blood group in clinical. 【Methods】 ABO and Rh blood group typing, absorption and elution test, and gene sequencing were performed to confirm the ambiguous blood group. 【Results】 The sample was identified by absorption and elution test and molecular biological method to be Ael subtype, and was named ABO*AEL.05/ABO*O.01.01 by ISBT. After family investigation, the proband and her second son share the same characteristic mutation site, and was named ABO*AEL.05/ABO*B.01.01 by ISBT. 【Conclusion】 Multiple blood group serological tests and molecular biology tests help to identify ABO subtypes, thus assuring the safety, scientificity and rationality of clinical blood transfusion.
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【Objective】 To analyze the serological characteristics and molecular mechanism of a novel B subtype allele 803delC. 【Methods】 ABO blood group was detected by serological method. Sequence-specific primer polymerase chain reaction (PCR-SSP) was used to detect ABO blood group genes. The coding region of exon 1-7 of ABO gene was detected by Sanger sequencing to determine the mutation site. 【Results】 Serological identification of patients was with forward O-type and reverse B-type. The result of PCR-SSP genotyping was A/O. There was A gene, which was not consistent with serological results. Further Sanger double-strand sequencing revealed that the C-base was deleted at position 803 of exon 7 on the basis of ABO*B. 01/ABO*O. 01.01. The mutation eventually leads to the amino acid substitution of p. Ala268Gly and p. Phe269Ser and the production of new open reading frame starting at position 269, with the new open reading frame No.20 amino acid being stop codon, resulted in the termination of B gene expression. Further single-strand sequencing of the ABO gene revealed that the mutation was located in the ABO*B. 01 gene. The mutation was submitted to the NCBI database with the number OR343908. 【Conclusion】 A new ABO allele leading to B variant has been found in Chinese population. Genetic detection can be used to identify the ambiguous blood group with discrepancy between forward and reverse blood grouping.
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Objective@#To investigate HIV-1 subtypes and drug resistance in HIV/AIDS patients who failed in antiretroviral therapy in Jinhua City, Zhejiang Province, so as to provide the basis for improving antiretroviral therapy strategy.@*Methods@#Totally 128 plasma samples of HIV/AIDS patients who failed in antiretroviral therapy (treatment for more than 6 months and viral load ≥1 000 copies/mL) from January 1 to November 30, 2023 were collected. After nucleic acid extraction and amplification, the sequences of HIV-1 pol genes were determined using first generation sequencing method, then submitted to HIV resistance database of Stanford University in the United States, and the subtypes, drug resistance mutations and drug resistance status of HIV-1 were analyzed.@*Results@#A total of 118 sequences of HIV/AIDS patients were obtained, including 94 males and 24 females (male to female ratio, 3.9︰1). There were 53 cases aged between 40 to 59 years, accounting for 44.92%. The main infection routes was heterosexual contact, with 92 cases accounting for 77.97%. The main HIV-1 gene subtypes were CRF07_BC and CRF01_AE, with 45 and 39 cases accounting for 38.14% and 33.05%, respectively. There were 75 cases found to have drug-resistant site mutations, with a mutation rate of 63.56%. The most common mutation sites were M184 and K103, with mutation rates of 29.66% and 28.81%, respectively. There were 58 cases with resistance to more than one drug, with a rate of 49.15%. The rates of resistance to non-nucleoside reverse transcriptase inhibitors (NNRTI), nucleotide reverse transcriptase inhibitors (NRTI) and protease inhibitors (PI) were 50.00%, 33.90% and 4.24%, respectively.@*Conclusion@#The HIV-1 gene subtypes of HIV/AIDS patients who failed in antiretroviral therapy in Jinhua City are mainly CRF07_BC and CRF01_AE, which are mainly resistant to NNRTI and NRTI.
摘要
Objective To investigate the prevalence of primary drug resistance among HIV-1 patients in Hubei Province from 2020 to 2022, and to provide corresponding basis and data support for HIV antiviral therapy (ART) in Hubei Province. Methods During 2020-2022, plasma samples of HIV-1 infected patients before ART were collected., Patients’ demographic data and baseline laboratory test data were also collected. HIV-1 pol region was amplified by in-house method for sub-type typing and drug-resistant mutation site analysis. Results The pol gene sequence was successfully amplified in 242 of 285 cases, with a success rate of 84.9%. CRF07_BC was the predominant HIV-1 sub-type, accounting for 47.11% (114/242), followed by CRF01_AE, accounting for 25.21% (61/242), sub-type B, accounting for 14.16% (35/242), and CRF55_01B, accounting for 4.13% (10/242). The primary resistance rate was 6.20% (15/242). The mutation site of nucleoside reverse transcriptase inhibitors (NRTIs) was mainly M184V, and the mutation sites of non-nucleoside reverse transcriptase inhibitors (NNRTIs) were mainly E138A/G/EG and V179E. These different mutation sites led to different degrees of drug resistance to 12 drugs. The incidence of drug resistance mutation of CRF55_01B sub-type was significantly higher than that of other sub-types. Conclusion The primary drug resistance rate of HIV-1 infected patients is at a slightly high level in Hubei Province, and close monitoring of primary drug resistance and mutation sites should be strengthened before ART, especially for CRF55_01B sub-type.
摘要
ABSTRACT The presence of genetic mutations in HIV poses a significant challenge, potentially leading to antiretroviral resistance and hampering therapeutic development. The Brazilian population has presented variations in the HIV envelope V3 loop gene, especially the GWGR motif. This motif has been linked to reduced transmission potential and slower CD4+ T cell decline. This study aimed to assess clinical outcomes in patients with HIV-1 infected with strains containing the GWGR motif compared with those without it during long-term cART. A cohort of 295 patients with HIV was examined for the GWGR motif presence in the V3 loop. A total of 58 samples showed the GWGR signature, while 237 had other signatures. Multifactorial analyses showed no significant differences in demographic characteristics, CD4+ cell count, AIDS progression, or mortality between GWGR carriers and others. However, the mean interval between the first positive HIV test and the initial AIDS-defining event was more than two times longer for women carrying the GWGR signature (p = 0.0231). We emphasize the positive impact of cART on HIV/AIDS treatment, including viral suppression, CD4+ cell preservation, and immune function maintenance. Although no significant differences were found during cART, residual outcomes reflecting adherence challenges were observed between diagnosis and the first AIDS-defining event. The previously described outcomes, highlighting statistically significant differences between individuals carrying the GPGR motif compared with those with the Brazilian GWGR motif, may be directly linked to the natural progression of infection before advancements in cART. Presently, these physicochemical aspects may no longer hold the same relevance.