A puerpera with a obstetric history of gravida 2, para 2, underwent blood typing due to the presence of agglutination reactions in her serum against all tested red blood cells. She was found to be blood type O and her RhD phenotype was identified as CcDEe through serological testing. The reaction agglutination intensity between her serum and 26 O-type blood cells from the panel was 2+. Whole genome sequencing was performed, yielding data on 4014 single nucleotide polymorphisms (SNPs) and 958 insertion/deletion (INDEL) loci across 50 genes responsible for encoding blood group systems. Among these, only a single SNP , rs72552713 was predicted to be a highly harmful variant, which is the c.376C>T variation in the ABCG2 gene encoding JR blood group antigen, leading to the premature stop codon (p.Gln126Ter). The c.376C>T variation has been named the ABCG2*01N.01 by the working party on Red Cell Immunogenetics and Blood Group Terminology of International Society of Blood Transfusion. The postpartum woman was found to have the Jr(a-) phenotype. Whole genome sequencing can accurately determine the antigens of blood group systems in some difficult specimens.

OBJECTIVE: To explore the molecular mechanism for an individual with Bweak subtype. METHODS: Serological methods were used to identify the proband's phenotype. In vitro enzyme activity test was used to determine the activity of B-glycosyltransferase (GTB) in her serum. The genotype was determined by PCR amplification and direct sequencing of exons 5 to 7 and flanking sequences of the ABO gene. T-A cloning technology was used to isolate the haploids. The primary physical and chemical properties and secondary structure of the protein were analyzed with the ProtParam and PSIPRED software. Three software, including PolyPhen-2, SIFT, and PROVEAN, was used to analyze the effect of missense variant on the protein. RESULTS: Serological results showed that the proband's phenotype was Bweak subtype with anti-B antibodies presented in her serum. In vitro enzyme activity assay showed that the GTB activity of the subject was significantly reduced. Analysis of the haploid sequence revealed a c.398T>C missense variant on the B allele, which resulted in a novel B allele. The 398T>C variant has caused a p.Phe133S substitution at position 133 of the GTB protein. Based on bioinformatic analysis, the amino acid substitution had no obvious effect on the primary and secondary structure of the protein, but the thermodynamic energy of the variant protein has increased to 6.07 kcal/mol, which can severely reduce the protein stability. Meanwhile, bioinformatic analysis also predicted that the missense variant was harmful to the protein function. CONCLUSION The weak expression of the Bweak subtype may be attributed to the novel allele of ABO*B.01-398C. Bioinformatic analysis is helpful for predicting the changes in protein structure and function.
Female ; Animals ; ABO Blood-Group System/genetics* ; Phenotype ; Genotype ; Exons ; Alleles

Objective: To investigate the status of hair loss and analyze the influencing factors among university students in Hangzhou City, so as to provide insights into the management of hair loss among university students. Methods: University students were recruited using a convenient sampling method from 4 universities in Hangzhou City in June 2021. The basic characteristics and life styles were collected using online questionnaire surveys. Self-reported hair loss was evaluated using the grading scales for loss of hair (Hamilton-Norwood scale for males and modified Ludwig scale for females), and factors affecting self-reported hair loss were identified among university students using the multivariable logistic regression model. Results: A total of 1 060 questionnaires were allocated, and 1 038 valid questionnaires were recovered, with an effective recovery rate of 97.92%. The respondents included 391 males ( 37.67% ) and 647 females ( 62.33% ), and 463 respondents ( 44.61% ) reported hair loss, including 431 students with mild hair loss ( 93.09% ). Multivariable logistic regression analysis showed that university students in their fourth or fifth years ( OR=1.721, 95%CI: 1.126-2.630 ), art specialty ( OR=0.411, 95%CI: 0.207-0.816 ), overweight or obesity (OR=1.685, 95%CI: 1.050-2.704), diet taste ( sweet: OR=2.131, 95%CI: 1.370-3.316; spicy: OR=1.510, 95%CI: 1.028-2.218; greasy: OR=3.023, 95%CI: 2.015-4.537 ), feeling nervous/anxious (occasionally: OR=1.891, 95%CI: 1.087-3.289; frequently: OR=2.487, 95%CI: 1.337-4.626 ), smoking ( occasionally: OR=1.906, 95%CI: 1.067-3.405; frequently: OR=1.983, 95%CI: 1.050-3.746), family history of hair loss ( OR=1.506, 95%CI: 1.075-2.110 ), perming/dyeing hair ( occasionally: OR=1.795, 95%CI: 1.280-2.517; frequently: OR=3.282, 95%CI: 1.736-6.204), self-perceived oily hair/scalp in the past three months (slightly increased: OR=1.980, 95%CI: 1.477-2.653; significantly increased: OR=5.347, 95%CI: 2.956-9.670) were factors affecting self-reported hair loss among university students. Conclusion The proportion of self-reported hair loss was 44.61% among university students in Hangzhou City, and hair loss was predominantly mild. A family history of hair loss, nervousness/anxiety, diet habits, smoking and frequency of perm/dyeing hair may affect hair loss among university students.

Background Di(2-ethylhexyl)phthalate (DEHP) and dibutyl phthalate (DBP) are representative environmental endocrine disruptors of phthalate esters (PAEs). Some studies have shown that PAEs exposure may have an impact on lipid metabolism. Objective To investigate the effects of DEHP and/or DBP on lipid metabolism in rats and their possible mechanisms of action. Methods Thirty-six weaned healthy SD male rats, 3 weeks old, weighing 50-70 g, were divided into four groups, i.e., a corn oil control group, a DEHP (750 mg·kg⁻¹) group, a DBP (500 mg·kg⁻¹) group, and a DEHP+DBP (750 mg·kg⁻¹+500 mg·kg⁻¹) group. The rats were exposed to DEHP and/or DBP by oral gavage for 8 weeks, and weighed once a week. The rats were anesthetized 24 h after the last dose, and blood was taken from the apical part of the heart. Serum high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), total cholesterol (TC), and triglyceride (TG) were detected. Liver tissues and perigenital adipose tissues were collected, weighed, and one portion of the tissues was fixed in 10% neutral formalin for pathomorphological observation, and another portion was used for mRNA detection of lipid metabolism-related genes such as Janus kinase 3 (JAK3), signal transducer and activator of transcription 5b (STAT5b), and peroxisome proliferator-activated receptor γ (PPARγ). Results During the DEHP and/or DBP exposure period, the rats in all groups were free to eat and drink without death or injury observed. Compared with the control group: The body weight gain in the DEHP+DBP group was lower at all time points from the 2nd week onwards (P<0.05); the liver organ coefficients of the DEHP and the DEHP+DBP groups were higher (P<0.05); the serum LDL-C levels in the DEHP and the DBP groups were higher (P<0.05). Compared with the DEHP+DBP group: The body weight gains in the DEHP group at the 2nd, 4th, 5th, and 8th weeks were higher (P<0.05), and the body weight gains in the DBP group were higher at all time points except the 1st week (P<0.05); the liver organ coefficients in the DEHP group and the DBP group were lower (P<0.05); the serum TG level in the DEHP group was higher(P<0.05), and the serum LDL-C levels in the DEHP and the DBP groups were higher (P<0.05). The pathomorphological results of liver tissues showed that the hepatocytes in the DEHP, DBP, and DEHP+DBP groups were disordered with loss of cord-like arrangement, swelling (suggesting change of cell proliferation), and presented bilirubin pigmentation. The pathomorphological results of rat perigenital adipose tissues showed had irregular alignment, sizes, and arrangement of adipocyte in the DEHP, DBP, and DEHP+DBP groups. The results of rat liver lipid metabolism-related gene mRNA levels showed that the liver JAK3, STAT5b, and PPARγ mRNA levels in the DEHP, DBP, and DEHP+DBP groups were lower than those in the control group (P<0.05); the rat liver PPARγ mRNA levels in the DEHP and DBP groups were lower than those in the DEHP+DBP group (P<0.05). Conclusion DEHP and/or DBP can inhibit the increase of body weight to varying degrees, induce inflammatory damage to liver tissues, and cause abnormal lipid metabolism in rats, and the associated mechanism may be related to inhibiting the activation of JAK3/STAT5b/PPARγ signaling pathway in rat liver tissues.

OBJECTIVE: To analyze the molecular characteristics of a recombinant allele of the ABO blood group. METHODS: The ABO phenotype was determined with the tube method. The coding regions of the ABO and FUT1 genes were analyzed by PCR-sequence based typing. The ABO alleles of the proband were determined by allele-specific primer sequencing. The full sequences of the ABO gene of the proband and her mother were determined through next generation sequencing. RESULTS: The red blood cells of the proband did not agglutinate with anti-H, and the sequence of the FUT1 gene was homozygous for c.551_552delAG.The proband was thereby assigned as para-Bombay. Bi-directional sequencing also found that she was heterozygous for c.261G/del,467C>T,c.526C>G,c.657C>T,c.703G>A,c.796C>A,c.803G>C and c.930G>A of the coding regions of the ABO gene. Allele-specific primer sequencing also found her to carry a ABO*A1.02 allele and a recombinant allele from ABO*O.01.01 and ABO*B.01. The recombination site was located between nucleotide c.375-269 and c.526, and the allele was maternally derived. CONCLUSION An recombinant allele of the ABO gene has been identified, which has originated from recombination of ABO*O.01.01 with the ABO*B.01 allele.
ABO Blood-Group System/genetics* ; Alleles ; Blood Grouping and Crossmatching ; Female ; Fucosyltransferases/genetics* ; Genotype ; Humans ; Phenotype ; Recombination, Genetic

OBJECTIVE: To explore the molecular basis for an individual with Bw subtype. METHODS: Routine serological reactions were used to determine the surface antigens of erythrocytes and antibodies in serum. PCR-sequence-based typing (PCR-SBT) was used to analyze the coding regions of the ABO gene and erythroid-specific regulatory element in its intron 1. Amplicons for exons 5 to 7 containing the variant site were subjected to TA cloning for the isolation of the haploid and verification of the sequence. The 3D structure of mutant protein was predicted with Pymol software. Changes of amino acid residues and structural stability were also analyzed. RESULTS: Serological assay showed that the individual had weakened B antigen and anti-B antibody in his serum. His genotype was determined as ABO*B.01/ABO*O.01.01. Sequencing of the entire coding region of the ABO gene identified an additional heterozygous c.734C/T variant. No variant was found in the erythroid-specific regulatory element of intron 1. Haploid cloning and isolation has obtained an ABO*O.01.01 allele and a ABO*B.01 allele containing a c.734T variant, which has led to substitution of Thr by Ile at position 245 in the functional center of glycosyltransferase. Based on the 3D structure of the protein, the residues binding with the mutation were unchanged, but the bonding distance between the hydrogens was changed with the amino acid substitution. Meanwhile, the connections with water molecules were increased. CONCLUSION The c.734C>T variant of the GTB gene can lead to an amino acid substitution in the functional center of the enzyme, which in turn may affect the stability of glycosyltransferase B protein and reduceits enzymatic activity.
ABO Blood-Group System/genetics* ; Alleles ; Exons/genetics* ; Genotype ; Glycosyltransferases/genetics* ; Humans ; Male ; Phenotype

Objective: To investigate the genotype and genetic characteristics of the pathogens associated with the epidemic outbreak of acute gastroenteritis(AGE) in Guangyuan city, Sichuan province. Methods: Eighteen stool samples and 15 anal swab samples from 4 AGE outbreaks were collected from Feb 2017 to May 2017. Norovirus (NoV) nucleic acid was detected by Real-time PCR method , and the positive samples were amplified by conventional reverse transcription-polymerase chain reaction (RT-PCR) and nucleotide sequencing. Results: Four AGE outbreaks were all caused by NoV. And 20 (60.6%) samples were positive for NoV GⅡ. Gene sequence comparison and phylogenetic analysis showed that 4 AGE outbreaks were all caused by GⅡ.P16/GⅡ.2 NoV. All the strains of GⅡ.P16/GⅡ.2 NoV shared high homology in nucleotides. One of the outbreaks was caused by GⅡ.P16/GⅡ.2 and GⅡ.P7/GⅡ.14 NoV co-infection. Conclusions The 4 outbreaks of AGE in Guangyuan city, Sichuan province were major caused by GⅡ.P16/GⅡ.2 NoV, meanwhile GⅡ.P16/GⅡ.2 and GⅡ.P7/GⅡ.14 NoV co-infection existed.

OBJECTIVETo explore the molecular basis of an individual with Bel variant of the ABO blood group.

METHODSThe ABO antigen and serum antibody of the individual were detected by serological method. All coding regions and flanking introns of the ABO gene were amplified with PCR and sequenced bidirectionally. The haplotypes of the individual were analyzed by cloning and sequencing. A three dimensional model of the mutant protein was constructed and analyzed.

RESULTSThe individual has expressed a very weak B antigen on its red blood cells by absorption and elution testing, which was identified as a Bel variant phenotype. The heterozygous sites in exon 6 (261del/G) and exon 7 (297A/G, 484del/G, 526C/G, 657C/T, 703G/A, 796C/A, 803G/C, 930G/A) of the coding region of the ABO gene were identified by direct sequencing. Haplotype analysis showed that the individual has carried an O01 allele and a novel B allele. The sequence of the novel B allele was identical to B101 except for a del G at nucleotide position 484 (484delG), which was nominated as B120 by the Blood Group Antigen Gene Mutation Database (dbRBC NCBI). The 484delG mutation of the B allele has led to a reading frame shift and created a premature terminal codon for the glycosyltransferase (GT) enzyme. Prediction of the 3D structure suggested that the GT enzyme has become an incomplete protein only with its N-terminal region.

CONCLUSIONThe 484delG mutation of the glycosyltransferase B gene has probably abolished or reduced the enzymatic activity and resulted in the Bel variant phenotype.

ABO Blood-Group System ; genetics ; Alleles ; Base Sequence ; Exons ; Female ; Genotype ; Glycosyltransferases ; genetics ; Humans ; Molecular Sequence Data ; Mutation ; Sequence Deletion

OBJECTIVETo compare the difference in the therapeutic effect on post-infectious cough differentiated as wind-cold retention in the lung between the combined therapy of scraping anddecoction and the simple application ofdecoction.

METHODSEighty patients were randomized into a combined therapy group and a Chinese herbal medicine group, 40 cases in each one. In the Chinese herbal medicine group, the oral administration ofdecoction was used. The main ingredients included,,,,,,,, etc., one dose a day, twice a day. In the combined therapy group, on the basis of the treatment as the Chinese herbal medicine group, scraping therapy was added and applied to the bladder meridian of foot-, the lung meridian of hand-, the conception vessel and the governor vessel, focusing on Tiantu (CV 22), Baihui (GV 20), Dazhui (GV 14), Feishu (BL 13), Fengmen (BL 12), Taiyuan (LU 9), Lieque (LU 7) and Fengchi (GB 20), once a week and one-week treatment as one session. Totally, the continuous two sessions were required in the two groups. The cough symptom score, cough remission time, relapse, TCM syndrome score, the score of Leicester cough questionnaire (LCQ), SP concentration in the supernatant of the induced sputum before and after treatment as well as clinical efficacy were observed in the two groups.

RESULTSThe cough symptom score, TCM symptom score and SP concentration in the supernatant of the induced sputum were all apparently reduced after treatment in the patients of the two groups (all<0.01). The scores in the combined therapy group were reduced in the higher amplitude as compared with those in the Chinese herbal medicine group (all<0.01). The total effective rate was 95.0% (38/40) in the combined therapy group, better than 87.5% (35/40) in the Chinese herbal medicine group (<0.05). Regarding the cough remission time and relapse rate, the results in the combined therapy group were better than those in the Chinese herbal medicine group[(5.3±1.2) d vs (7.4±1.5) d,<0.01; 0% (0/19) vs 62.5% (5/8),<0.01]. The scoreo of LCQ was all apparently improved in the patients of the two groups (both<0.01), and the score in the combined therapy group was higher than that in the Chinese herbal medicine group (<0.01).

CONCLUSIONSScraping therapy combined withdecoction and the simple application ofdecoction all relieve the symptoms of post-infectious cough and improves the living quality. The therapeutic effects of the combined therapy are superior to the oral administration ofdecoction.

OBJECTIVETo explore the function of a novel nonsense mutation c.1476G>A of ITGB3 gene using an in vitro expression system.

METHODSAn eukaryotic expression vector containing ITGB3 c.1476G>A cDNA was generated by site-directed mutagenesis and transformed into E.coli. Plasmid DNA was extracted and sequenced to confirm the target mutations. Wild-type and mutant recombination plasmids were transfected into Chinese hamster ovarian cancer (CHO) cells by nonliposome method, and the stable expression cells were harvested by G418 screening. The ITGB3 gene mRNA transcription and GPIIIa expression level in CHO cells were detected with real-time quantitative PCR, Western blotting and flow cytometry, respectively.

RESULTSThe eukaryotic expression vectors of wild ITGB3 cDNA and c.1476G>A mutant were successfully constructed. CHO cells with stable expression were obtained after transfection and screening. Compared with the wild-type transfected cells, the amount of CD61 antigen expression was 37% and mRNA transcription level was only 6% in the mutant-transfected cells. Full length GPIIIa protein was found only in the stably wild-type-transfected cells, but not in mutant-transfected cells by Western blotting analysis.

CONCLUSIONThe ITGB3 c.1476G>A mutation can decrease the transcription level and further affect GPIIIa synthesis and CD61 antigen expression.

Animals ; Base Sequence ; Blood Platelets ; cytology ; metabolism ; CHO Cells ; Cloning, Molecular ; Codon, Nonsense ; genetics ; Cricetinae ; Cricetulus ; Humans ; Integrin beta3 ; genetics ; metabolism ; Molecular Sequence Data ; Plasmids ; genetics ; metabolism ; Point Mutation