[Objective] To conduct serological identification and molecular mechanism study on a ambiguous ABO blood group. [Methods] Standard serological techniques were used for the forward and reverse typing of ABO blood type. ABO gene coding and regulatory regions were analyzed by PCR after DNA extraction. Monoclonal sequencing was used to detect the haplotypes of the DNA sequence, and bioinformatics analysis was applied to predict the possible translation outcomes of the mutated DNA sequence. [Results] The sample’s red blood cells showed mixed field agglutination with anti-A, and the serum agglutinated with B cells, exhibiting serological characteristics of subtype A. Direct sequencing and monoclonal sequencing analysis of the ABO gene confirmed one allele as O02, the other had a c.27delC mutation compared with A102, which could cause the translation sequence to terminate prematurely at the 19th amino acids. Analysis and prediction suggested that the mutation might affect the function of the transferase through mechanisms such as shifting the initiation codon, altering the reading frame and affecting the splice sites. [Conclusion] This case is a rare A subtype caused by the c.27delC variation, and the impact on the glycosyltransferase may involve multiple mechanisms, which require further research and exploration.

Objective: To perform serological identification and molecular analysis of four samples with antibodies against high-frequency antigens, and to track the condition of newborns after delivery. Methods: Blood group serological tests were conducted using tube method, and unexpected antibody screening and identification were performed using polybrene and human globulin card. Gene haplotype analysis of blood group system was performed using PacBio third-generation sequencing. The DNA mutations were confirmed through Sanger sequencing. Results: Four samples showed normal blood types in common blood type systems. However, they were positive in all unexpected antibody screening and identification, with negative direct antiglobulin tests results. Third-generation sequencing revealed 3 cases of c.376C>T homozygous mutation and 1 case of c.421C>A homozygous mutation in the ABCG2 gene. Three pregnant women gave birth to four children, all of whom developed hyperbilirubinemia, accompanied by decreased red blood cell count and normal or low hemoglobin concentration. Conclusion: Four samples were obtained from individuals with the rare Jr(a-) blood group. Immunization during pregnancy led to the production of anti-Jr antibody, which may contribute to hyperbilirubinemia in newborns.

Objective To investigate the effects of long-term exposure to three new types of light emitting diode (LED) sources on the behavior, learning, and memory of rats. Methods A total of 160 specific pathogen-free SD rats were divided into eight groups as followed, trichromatic fluorescent lamps color temperature control group, violet-chip full-spectrum white LED group, blue-chip white LED group, and blue-chip full-spectrum white LED group based on the light sources types, with color temperature of 4 000 K and 6 500 K groups in each group using the 4×2 factorial design. There were 20 rats in each group, with half of the rats were males and half females. Rats were exposed to artificial lighting, and the illumination was set at 750 lx. The rats in each group were exposed to different lighting environments for 12 hours per day for 24 weeks. The open-field and step-down tests were conducted in rats after 24 weeks exposure, followed by sacrifice of rats and measurement of organ coefficients. Differences in body weight, organ coefficients, and neurobehavioral indexes of rats in different groups were compared. Results The spleen coefficient of female rats decreased in blue-chip white LED of 6 500 K color temperature group, and the liver coefficient of male rats decreased in the violet-chip full-spectrum white LED of 4 000 K color temperature, blue-chip full-spectrum white LED of 4 000 K color temperature, and blue-chip full-spectrum white LED of 6 500 K color temperature groups, compared with the same-sex rats in trichromatic fluorescent lamps with same-color temperature control group (all P<0.05). The result of different types of light sources compared in the open-field test showed that the index of total distance and movement speed of female rats in the blue-chip full-spectrum white LED group were lower than those in the other three groups, and the time cost to the central area was longer than that in the blue-chip white LED group and the violet-chip full-spectrum white LED group (all P<0.05). The total distance and movement speed of male rats in the blue-chip full-spectrum white LED group were longer or higher than those in the violet-chip full-spectrum white LED group (all P<0.05). Based on the comparison of color temperature, the time and total distance of male rats in 6 500 K color temperature group were lower than that in the 4 000 K color temperature group (both P<0.05). In the step-down test, both male and female rats in the blue-chip full-spectrum white LED group made more errors compared with other three groups with the same gender (all P<0.05). Conclusion Based on the experimental conditions of this study, the blue-chip full-spectrum white light LED affects behavior, learning and memory of the rats, and trichromatic fluorescent lamp has the lowest effect on neurobehavior. The color temperature also affects behavior of the rats, and high color temperature has higher risk.

The density and composition of lymphocytes infiltrating colon tumors serve as predictive factors for the clinical outcome of colon cancer.Our previous studies highlighted the potent anti-cancer properties of the principal compounds found in Garcinia yunnanensis(YTE-17),attributing these effects to the regu-lation of multiple signaling pathways.However,knowledge regarding the mechanism and effect of YTE-17 in the prevention of colorectal cancer is limited.In this study,we conducted isobaric tags for relative and absolute quantification(iTRAQ)analysis on intestinal epithelial cells(IECs)exposed YTE-17,both in vitro and in vivo,revealing a significant inhibition of the Wnt family member 5a(Wnt5a)/c-Jun N-terminal kinase(JNK)signaling pathway.Subsequently,we elucidated the influence and mechanism of YTE-17 on the tumor microenvironment(TME),specifically focusing on macrophage-mediated T helper 17(Th17)cell induction in a colitis-associated cancer(CAC)model with Wnt5a deletion.Additionally,we performed the single-cell RNA sequencing(scRNA-seq)on the colonic tissue from the Wnt5a-deleted CAC model to characterize the composition,lineage,and functional status of immune mesenchymal cells during different stages of colorectal cancer(CRC)progression.Remarkably,our findings demon-strate a significant reduction in M2 macrophage polarization and Th17 cell phenotype upon treatment with YTE-17,leading to the restoration of regulatory T(Treg)/Th17 cell balance in azoxymethane(AOM)/dextran sodium sulfate(DSS)model.Furthermore,we also confirmed that YTE-17 effectively inhibited the glycolysis of Th17 cells in both direct and indirect co-culture systems with M2 macrophages.Notably,our study shed light on potential mechanisms linking the non-canonical Wnt5a/JNK signaling pathway and well-established canonical β-catenin oncogenic pathway in vivo.Specifically,we proposed that Wnt5a/JNK signaling activity in IECs promotes the development of cancer stem cells with β-catenin activity within the TME,involving macrophages and T cells.In summary,our study undergoes the po-tential of YTE-17 as a preventive strategy against CRC development by addressing the imbalance with the immune microenvironment,thereby mitigating the risk of malignancies.

AIM:To explore the impact of histidine triad nucleotide-binding protein 2(HINT2)on atheroscle-rosis(AS),and to elucidate its underlying mechanisms.METHODS:Peripheral blood mononuclear cells(PBMCs)were isolated from patients with chest pain and induced to differentiate into macrophages for assessing HINT2 expression.In vitro,RAW264.7 mouse macrophages were cultured to evaluate inflammatory cytokine levels and cell death in superna-tants.An apolipoprotein E gene knockout(apoE-/-)mouse model of AS was developed to analyze plaque formation,lipid metabolism and macrophage foaminess in the aortic root.Pyroptosis-related protein expression in cultured RAW264.7 cells and the aorta of apoE-/-mice was determined by RT-qPCR and Western blot.RESULTS:Significantly reduced HINT2 expression was observed in PBMCs from patients with acute coronary syndrome,which was negatively correlated with pro-inflammatory and positively correlated with anti-inflammatory serum factors,suggesting HINT2's potential role in mitigating AS-related inflammation.In vitro experiments demonstrated that HINT2 overexpression inhibited lipid accumu-lation and foam cell formation in macrophages induced by oxidized low-density lipoprotein(ox-LDL).It also reduced se-cretion of inflammatory cytokines,including interleukin-6(IL-6),IL-1β,IL-18 and tumor necrosis factor-α,and de-creased cell death and pyroptosis-related protein expression.In vivo experiments in apoE-/-mice confirmed that HINT2 overexpression lessened plaque burden in the aortic root,reduced macrophage foaminess,and inhibited the expression of pyroptosis-related proteins such as nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3),caspase-1,gasdermin D(GSDMD),IL-1β and IL-18.CONCLUSION:HINT2 potentially inhibits ox-LDL-induced macrophage pyroptosis,attenuates inflammatory responses in AS,and may slow the progression of this disease.

【Objective】 To conduct serological identification and molecular study of two patients with ABO ambiguous blood group. 【Methods】 The serological tests were conducted by the tube method. DNA direct sequencing was performed to analyze the exons and transcriptional regulatory regions of the ABO gene. TA clone sequencing was performed to confirm the mutation sites of the haploid. DeepTMHMM was used for transmembrane region prediction and analysis. 【Results】 Both samples showed weak agglutination with anti-A in forward typing and the presence of anti-A antibodies in reverse typing. ABO gene analysis confirmed 1A>G and 467C>T mutations of A101 gene, indicating the Aw43 allele.DeepTMHMM analysis showed that 1A>G mutation shift back the translation start site, which would affect the transmembrane structure seriously. 【Conclusion】 The two cases of ABO ambiguous blood group were with Aw43 alleles. The 1A>G mutation affected the transmembrane region and subsequently altered the glycoprotein structure, resulting in impaired enzyme function.

Objective:Clarify the correlation between the Centralized Drug Volume-based Purchasing Policy,changes in physical&mental health status,and health inequalities among the older adults,as well as exploring whether this correlation differs between urban and rural areas so as to determine the role played by the total cost of healthcare.Methods:Based on CFPS 2018 and 2020,DID and PSM-DID were used to verify the impact of the Centralized Drug Volume-based Purchasing Policy on physical&mental health status and health inequalities of the elderly in urban and rural China.Three-step method and Sobel test were used to verify the mediating effect of total medical costs.Results:The pilot effect of the Centralized Drug Volume-based Purchasing Policy was first realized among the urban elderly population,promoting their physical and mental health through the intermediary mechanism of reducing the total medical costs,but for the time being did not show a significant correlation with health inequalities within this group.Among the rural elderly population,none of the three dependent variables was significant(P＞0.1).Conclusion:The underlying logic of the Centralized Drug Volume-based Purchasing Policy is right,but its future reform should focus on the rural elderly group,and consider the fairness of the effect within the group and between urban and rural areas,so as to improve the health level of the whole society in the longer term.

Objective:Clarify the correlation between the Centralized Drug Volume-based Purchasing Policy,changes in physical&mental health status,and health inequalities among the older adults,as well as exploring whether this correlation differs between urban and rural areas so as to determine the role played by the total cost of healthcare.Methods:Based on CFPS 2018 and 2020,DID and PSM-DID were used to verify the impact of the Centralized Drug Volume-based Purchasing Policy on physical&mental health status and health inequalities of the elderly in urban and rural China.Three-step method and Sobel test were used to verify the mediating effect of total medical costs.Results:The pilot effect of the Centralized Drug Volume-based Purchasing Policy was first realized among the urban elderly population,promoting their physical and mental health through the intermediary mechanism of reducing the total medical costs,but for the time being did not show a significant correlation with health inequalities within this group.Among the rural elderly population,none of the three dependent variables was significant(P＞0.1).Conclusion:The underlying logic of the Centralized Drug Volume-based Purchasing Policy is right,but its future reform should focus on the rural elderly group,and consider the fairness of the effect within the group and between urban and rural areas,so as to improve the health level of the whole society in the longer term.

Objective:Clarify the correlation between the Centralized Drug Volume-based Purchasing Policy,changes in physical&mental health status,and health inequalities among the older adults,as well as exploring whether this correlation differs between urban and rural areas so as to determine the role played by the total cost of healthcare.Methods:Based on CFPS 2018 and 2020,DID and PSM-DID were used to verify the impact of the Centralized Drug Volume-based Purchasing Policy on physical&mental health status and health inequalities of the elderly in urban and rural China.Three-step method and Sobel test were used to verify the mediating effect of total medical costs.Results:The pilot effect of the Centralized Drug Volume-based Purchasing Policy was first realized among the urban elderly population,promoting their physical and mental health through the intermediary mechanism of reducing the total medical costs,but for the time being did not show a significant correlation with health inequalities within this group.Among the rural elderly population,none of the three dependent variables was significant(P＞0.1).Conclusion:The underlying logic of the Centralized Drug Volume-based Purchasing Policy is right,but its future reform should focus on the rural elderly group,and consider the fairness of the effect within the group and between urban and rural areas,so as to improve the health level of the whole society in the longer term.

Objective:Clarify the correlation between the Centralized Drug Volume-based Purchasing Policy,changes in physical&mental health status,and health inequalities among the older adults,as well as exploring whether this correlation differs between urban and rural areas so as to determine the role played by the total cost of healthcare.Methods:Based on CFPS 2018 and 2020,DID and PSM-DID were used to verify the impact of the Centralized Drug Volume-based Purchasing Policy on physical&mental health status and health inequalities of the elderly in urban and rural China.Three-step method and Sobel test were used to verify the mediating effect of total medical costs.Results:The pilot effect of the Centralized Drug Volume-based Purchasing Policy was first realized among the urban elderly population,promoting their physical and mental health through the intermediary mechanism of reducing the total medical costs,but for the time being did not show a significant correlation with health inequalities within this group.Among the rural elderly population,none of the three dependent variables was significant(P＞0.1).Conclusion:The underlying logic of the Centralized Drug Volume-based Purchasing Policy is right,but its future reform should focus on the rural elderly group,and consider the fairness of the effect within the group and between urban and rural areas,so as to improve the health level of the whole society in the longer term.