Objective: To identify the phenotypes, antibodies and explore the molecular mechanisms of a patient who carries antibodies to RBC high-frequency antigens and his family members. Methods: The antibody identification test was performed for the proband by serological methods, and targeted NGS was subsequently used to detect mutations that occurred in blood group genes. Blood samples were collected from the proband and his family members. Sanger sequencing was used to verify the mutation of the XK gene. The expression of Kell blood group antigens was detected by serological methods and flow cytometry. K cells were used to detect the antibody specificity of the proband. The morphology of red blood cells was detected by the scanning electron microscopy. The serum creatine kinase levels of the proband and his family members were analyzed by colorimetric methods. Results: The results of the antibody identification test suggested that the proband might have antibodies to high-frequency antigens. NGS results suggested a homozygous mutation (c. 107G>A) in exon 1 of the XK gene in the proband, resulting in a truncated XK protein. The Sanger sequencing results of the proband were consistent with the NGS results, and the mutation was not found in other family members. The expression of Kell blood group antigens of the proband was not found by serological methods and flow cytometry. The results of the antibody specificity test showed that the proband had anti-Km antibodies. Spike-like changes were identified on red blood cells, and serum creatine kinase level was elevated in the proband. Conclusion: In this study, the McLeod phenotype caused by homozygous mutation (c. 107G>A) of XK gene was identified in Chinese individuals for the first time by the phenotype and molecular mechanism studies. The results of genotyping and phenotyping suggested that the McLeod phenotype caused by the mutation was compatible with the phenotypes of McLeod and K .

Objective:To investigate the effects of 3 adhesives on the bond force and durability of polished and glazed zirconia ceram-ics to orthodontic metal brackets respectively.Methods:Universal adhesives,Single Bond Universal(SBU)and Prime&Bond Universal(PBU)were respectively used to bond polished and glazed zirconia to metal braces of maxillary central incisors using TransbondTM MIP(TM)as the control.The shear bond strength(SBS),the fracture morphology and adhesive residual index(ARI)were examed after wa-ter bath or water bath-thermal cycling storage.Results:The adhesive(P＜0.001)and storage conditions(P＜0.001)significantly af-fected the shear bond strength of zirconia to brackets.There was no significant difference between the polished or glazed groups(P=0.09).SBU showed the stronger SBS and lower ABI,there were significant differences in ARI scores among the 3 cements(P＜0.001).Conclusion:SBU may have better bonding performance than PBU and TM in the orthodontic bonding of polished or glazed zir-conia surfaces to the zirconia ceramics.

Objective:To analyze the risk factors of unfavourable treatment outcomes in patients with rifampicin-sensitive pulmonary tuberculosis（PTB）.Methods:Clinical data of 726 patients with rifampicin-sensitive PTB admitted in Quzhou Hospital Affiliated to Wenzhou Medical University from January 2020 to December 2021 were retrospectively analyzed，including 628 cases with favourable treatment outcomes（favourable group）and 98 cases with unfavourable treatment outcomes（unfavourable group）. After excluding the lost to follow-up cases and non-tuberculosis-related death cases，there were 659 survival cases（survival group）and 16 cases of tuberculosis-related death（fatal group）. Univariate and multivariate Logistic regression were used to analyze the related risk factors.Results:Among 98 cases with unfavourable treatment outcomes，42 cases died from non-tuberculosis causes，32 cases lost follow-up，16 cases died from tuberculosis and 8 cases had failed treatment. Multivariate Logistic regression analysis showed that age≥60（ OR=2.515，95% CI 1.461-4.330），retreatment of tuberculosis（ OR=2.502，95% CI 1.255-4.989），chronic obstructive pulmonary disease（ OR=2.796，95% CI 1.607-4.865），respiratory failure（ OR=6.373，95% CI 2.320-17.506），hypoproteinemia（ OR=3.149，95% CI 1.876-5.286）and elevated C-reactive protein（ OR=1.007，95% CI 1.002-1.012）were independent risk factors for unfavorable treatment outcomes in rifampicin-sensitive PTB patients（all P<0.01）；retreated pulmonary tuberculosis（ OR=4.542，95% CI 1.182-17.449），complicated respiratory failure（ OR=14.186，95% CI 3.038-66.252）and body mass index<18.5 kg/m 2（ OR=5.179，95% CI 1.333-20.119）were independent risk factors for poor prognosis in rifampicin-sensitive PTB patients（ P<0.05 or <0.01）. Conclusion:The older age，low BMI，hypoproteinemia，elevated C-reactive protein，comorbidities and retreatment are risk factors for unfavorable treatment outcomes and poor prognosis in patients with rifampicin-sensitive PTB.

Insomnia disorder is a common disease in acupuncture clinics.There is a tendency for the theory to solidify and lag behind the clinic.In this paper,we examined insomnia disorder from the perspective of acupuncture and moxibustion theory.Based on the general law of acupuncture and moxibustion to recognize the disease,we concerned that the basic understanding of the disease has commonality between ancient and modern medical practitioners,and the theory can be deconstructed as follows:the theory of"yin and yang"is the starting point and the root of the theory,the theory of"spirit regulars the sleep"is the directly related theory,the theory of internal organs is the basis of the disease mechanism,the theory of meridians and acupoints is the key to diagnosis and treatment,and the theory of acupuncture and moxibustion is the key to treatment.The improvement and development of the theory of acupuncture for insomnia disorder can be carried out on this basis.

Insomnia disorder is a common disease in acupuncture clinics.There is a tendency for the theory to solidify and lag behind the clinic.In this paper,we examined insomnia disorder from the perspective of acupuncture and moxibustion theory.Based on the general law of acupuncture and moxibustion to recognize the disease,we concerned that the basic understanding of the disease has commonality between ancient and modern medical practitioners,and the theory can be deconstructed as follows:the theory of"yin and yang"is the starting point and the root of the theory,the theory of"spirit regulars the sleep"is the directly related theory,the theory of internal organs is the basis of the disease mechanism,the theory of meridians and acupoints is the key to diagnosis and treatment,and the theory of acupuncture and moxibustion is the key to treatment.The improvement and development of the theory of acupuncture for insomnia disorder can be carried out on this basis.

Objective:To analyze the risk factors of unfavourable treatment outcomes in patients with rifampicin-sensitive pulmonary tuberculosis（PTB）.Methods:Clinical data of 726 patients with rifampicin-sensitive PTB admitted in Quzhou Hospital Affiliated to Wenzhou Medical University from January 2020 to December 2021 were retrospectively analyzed，including 628 cases with favourable treatment outcomes（favourable group）and 98 cases with unfavourable treatment outcomes（unfavourable group）. After excluding the lost to follow-up cases and non-tuberculosis-related death cases，there were 659 survival cases（survival group）and 16 cases of tuberculosis-related death（fatal group）. Univariate and multivariate Logistic regression were used to analyze the related risk factors.Results:Among 98 cases with unfavourable treatment outcomes，42 cases died from non-tuberculosis causes，32 cases lost follow-up，16 cases died from tuberculosis and 8 cases had failed treatment. Multivariate Logistic regression analysis showed that age≥60（ OR=2.515，95% CI 1.461-4.330），retreatment of tuberculosis（ OR=2.502，95% CI 1.255-4.989），chronic obstructive pulmonary disease（ OR=2.796，95% CI 1.607-4.865），respiratory failure（ OR=6.373，95% CI 2.320-17.506），hypoproteinemia（ OR=3.149，95% CI 1.876-5.286）and elevated C-reactive protein（ OR=1.007，95% CI 1.002-1.012）were independent risk factors for unfavorable treatment outcomes in rifampicin-sensitive PTB patients（all P<0.01）；retreated pulmonary tuberculosis（ OR=4.542，95% CI 1.182-17.449），complicated respiratory failure（ OR=14.186，95% CI 3.038-66.252）and body mass index<18.5 kg/m 2（ OR=5.179，95% CI 1.333-20.119）were independent risk factors for poor prognosis in rifampicin-sensitive PTB patients（ P<0.05 or <0.01）. Conclusion:The older age，low BMI，hypoproteinemia，elevated C-reactive protein，comorbidities and retreatment are risk factors for unfavorable treatment outcomes and poor prognosis in patients with rifampicin-sensitive PTB.

Objective:To explore the relationship between workplace procrastination and illegitimate tasks in-kindergarten teachersand the role of work disengagement and coworker support in their relationship.Methods:A to-tal of 245 kindergarten teachers were selected from 3 cities in Zhejiang Province.They were assessed with the Workplace Procrastination Scale(WPS),Bern Illegitimate Tasks Scale(BITS),Work Disengagement Scale(WDS),Colleague Support Scale(CSS).The models were tested by using Process macro for SPSS,and non-para-metric percentile bootstrap method was used to analyze the mediating effect and moderating effect.Results:There were significant differences in the total scores of workplace procrastination among kindergarten teachers in different marital status,age,teaching age,education level,teaching gradeand kindergarten level(Ps＜0.05).Work disengage-ment played a significant mediating role between workplace procrastination and illegitimate tasks(indirect effect=0.26,95％CI:0.16-0.37).Coworker support played a significant moderating role in the impact of illegitimate tasks on work disengagement(simple slope=0.72,0.39;P＜0.001).Conclusion:It suggests that workplace pro-crastination is related to illegitimate tasksin kindergarten teachers.Work disengagement plays a mediating role in their relationship,and coworker support plays a moderating role in the first half of this mediating role.

【Objective】 To explore the application of matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) in the genotyping of difficult blood typing samples, and to provide evidence for clinical blood transfusion. 【Methods】 Three ambiguous blood group samples, submitted to Shanghai Blood Center by Shanghai regional hospitals, were studied, of which Sample1 included the proband and his parents. Serological methods were used to perform blood group typing, direct antibody test, unexpected antibody screening and identification test. Blood group genotyping was performed by using the MALDI-TOF MS detection systeme stablished in our laboratory. Sanger sequencing was used to confirm gene mutation sites, and serological or flow methods were used to verify specific samples′ phenotype. 【Results】 Serological results indicated the existence of antibodies against high frequency antigens in sample 1 (including proband and her mother), 2 and 3. The genotyping results of MALDI-TOF MS showed that the proband of sample 1 was Di(a+ b+ ), her father was Di(a-b+ ), her mother was Di(a+ b-), sample 2 was p, and sample 3 was Jr(a-). Sequencing results of three samples were consistent with mass spectrometry typing results. Serological results showed that sample 2 had a p phenotype. The flow cytometry results suggested that sample 3 had a Jr(a-) phenotype. 【Conclusion】 For the first time, we applied MALDI-TOF MS technology to blood type genotyping of ambiguous clinical samples in China. Compared with other genotyping methods such as PCR-SSP, MALDI-TOF MS has the advantages of rapid detection, high throughput and high specificity, which would contribute to identification of difficult blood typing samples in the future, as well as rare blood group screening.

OBJECTIVETo investigate whether miR-492 is involved in the post-transcriptional regulation of OK blood group antigen expression on red blood cells.

METHODSTwo 3'-UTR fragments of the BSG gene were synthesized with a chemical method, which respectively encompassed the BSG rs8259 TT or BSG rs8259 AA sites. The fragments were added with Xho I and Not I restriction enzyme cutting sites at both ends and cloned into a pUC57 vector, which in turn was constructed into a psiCHECK-2 vector and verified by sequencing. K562 cells were transfected with various combinations of miR-492 mimic and constructed psiCHECK2-BSG-T or psiCHECK2-BSG-A recombinant plasmid. A blank control group was set up. Each transfection experiment was repeated three times. The activity of Renilla reniformis luciferase was determined and normalized with that of firefly luciferase, and detected with a dual-luciferase reporter assay system. The data were subjected to statistical analysis.

RESULTSThe sequencing results confirmed that the recombinant psiCHECK2 plasmids containing the BSG rs8259 TT or rs8259 AA sites were constructed successfully. The results of dual-luciferase report gene detection showed that the miR-492 mimic could significantly inhibit psiCHECK2-BSG-T at a concentration over 100 nmol/L. However, it could not inhibit psiCHECK-BSG-A.

CONCLUSIONmiR-492 may be involved in the regulation of OK antigen expression on red blood cells with the BSG rs8259 TT genotype.

Basigin ; genetics ; Blood Group Antigens ; genetics ; Erythrocytes ; immunology ; Gene Expression Regulation ; Genotype ; Humans ; MicroRNAs ; physiology

OBJECTIVETo screen rare blood groups Fy(a-), s-, k-, Di(b-) and Js(b-) in an ethnic Zhuang population.

METHODSSequence-specific primers were designed based on single nucleotide polymorphism (SNP) sites of blood group antigens Fy(b) and s. A specific multiplex PCR system I was established. Multiplex PCR system II was applied to detect alleles antigens Di(b), k, Js(b)1910 and Js(b) 2019 at the same time. The two systems was were used to screen for rare blood group antigens in 4490 randomly selected healthy donors of Guangxi Zhuang ethnic origin.

RESULTSWe successfully made the multiplex PCR system I. We detected the rare blood group antigens using the two PCR system. There are five Fy(a-), three s(-), two Di(b-) in 4490 Guangxi zhuang random samples. The multiplex PCR system I has achieved good accuracy and stability. With multiplex PCR systems I and II, 4490 samples were screened. Five Fy(a-), three s(-) and two Di(b-) samples were discovered.

CONCLUSIONMultiplex PCR is an effective methods, which can be used for high throughput screening of rare blood groups. The rare blood types of Guangxi Zhuang ethnic origin obtained through the screening can provide valuable information for compatible blood transfusion. Through screening we obtained precious rare blood type materials which can be used to improve the capability of compatible infusion and reduce the transfusion reactions.

Blood Group Antigens ; genetics ; Duffy Blood-Group System ; genetics ; Genotype ; Humans ; Multiplex Polymerase Chain Reaction ; methods ; Receptors, Cell Surface ; genetics