Objective::This study aimed to elucidate the effect of the lipopolysaccharides/toll-like receptor 4 (LPS/TLR4) pathway on early atherosclerosis (AS) development and its associated changes in fecal metabolites, thereby providing an experimental foundation for strategies to prevent and treat early AS.Methods::Twelve Tibetan miniature pigs aged 4-5 months were divided into normal control (NC) group and AS group (6 pigs in each). The group assignment was primarily based on body weight; Secondary criteria, including glucose, lipid profiles, and inflammatory indices, were considered to ensure balanced baseline characteristics between the 2 groups (all P > 0.05). AS group received a high-fat diet for 16 weeks to establish an AS model, while the NC group received a normal diet. Subsequently, serum levels of lipids and various inflammation and oxidative stress markers were measured. Pathological changes in the aorta and colon tissue, LPS/TLR4 pathway-associated protein expressions in the aorta, as well as occludin and zonula occludens-1 in the colon were also assessed. Proton nuclear magnetic resonance spectra technology was employed for the metabolomic analysis of fecal extracts. Results::The lipid metabolism was disrupted in AS group, with significantly higher total cholesterol, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol levels ((12.24 ± 5.24) mmol/L vs. (1.86 ± 0.27) mmol/L, P = 0.004,6; (2.39 ± 0.50) mmol/L vs. (0.83 ± 0.07) mmol/L, P = 0.000,5; (6.94 ± 2.87) mmol/L vs. (0.77 ± 0.18) mmol/L, P = 0.003,3), as compared to that in NC group. Serum factors, including LPS, tumor necrosis factor-α, and malondialdehyde levels of AS group were significantly higher than that of NC group ((1,230.00 ± 192.70) EU/L vs. (695.70 ± 213.70) EU/L), P = 0.001,1; (424.20 ± 176.90) ng/L vs. (51.20 ± 26.61) ng/L, P = 0.023,5; (3.60 ± 0.77) nmol/mL vs. (2.62 ± 0.21) nmol/mL, P = 0.025,4). Pathological evaluations revealed prominent lipid deposition area in the aortic arch, thoracic aorta, and abdominal aorta of the AS group compared with that of the NC group (4.17% ± 2.30% vs. 0, P = 0.006,7; 6.23% ± 2.95% vs. 0, P = 0.003,6; 3.78% ± 2.18% vs. 0, P = 0.008,1). TLR4, nuclear factor kappa-B p65, and tumor necrosis factor-α expression in the aorta tissue of the AS group were upregulated, whereas occludin and zonula occludens-1 expression in colon tissues was downregulated. Additionally, metabolomics identified significant differences in 21 metabolites in the feces of the AS group compared to the NC group, with further analysis linking these differences to amino acid metabolism. Conclusions::The Tibetan miniature pig model of early AS induced by high-fat intake displayed pronounced chronic inflammation. Preliminary findings suggest that the underlying mechanisms may be associated with the LPS/TLR4 pathway and intestinal metabolic disorders.

Objective::This study aimed to elucidate the effect of the lipopolysaccharides/toll-like receptor 4 (LPS/TLR4) pathway on early atherosclerosis (AS) development and its associated changes in fecal metabolites, thereby providing an experimental foundation for strategies to prevent and treat early AS.Methods::Twelve Tibetan miniature pigs aged 4-5 months were divided into normal control (NC) group and AS group (6 pigs in each). The group assignment was primarily based on body weight; Secondary criteria, including glucose, lipid profiles, and inflammatory indices, were considered to ensure balanced baseline characteristics between the 2 groups (all P > 0.05). AS group received a high-fat diet for 16 weeks to establish an AS model, while the NC group received a normal diet. Subsequently, serum levels of lipids and various inflammation and oxidative stress markers were measured. Pathological changes in the aorta and colon tissue, LPS/TLR4 pathway-associated protein expressions in the aorta, as well as occludin and zonula occludens-1 in the colon were also assessed. Proton nuclear magnetic resonance spectra technology was employed for the metabolomic analysis of fecal extracts. Results::The lipid metabolism was disrupted in AS group, with significantly higher total cholesterol, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol levels ((12.24 ± 5.24) mmol/L vs. (1.86 ± 0.27) mmol/L, P = 0.004,6; (2.39 ± 0.50) mmol/L vs. (0.83 ± 0.07) mmol/L, P = 0.000,5; (6.94 ± 2.87) mmol/L vs. (0.77 ± 0.18) mmol/L, P = 0.003,3), as compared to that in NC group. Serum factors, including LPS, tumor necrosis factor-α, and malondialdehyde levels of AS group were significantly higher than that of NC group ((1,230.00 ± 192.70) EU/L vs. (695.70 ± 213.70) EU/L), P = 0.001,1; (424.20 ± 176.90) ng/L vs. (51.20 ± 26.61) ng/L, P = 0.023,5; (3.60 ± 0.77) nmol/mL vs. (2.62 ± 0.21) nmol/mL, P = 0.025,4). Pathological evaluations revealed prominent lipid deposition area in the aortic arch, thoracic aorta, and abdominal aorta of the AS group compared with that of the NC group (4.17% ± 2.30% vs. 0, P = 0.006,7; 6.23% ± 2.95% vs. 0, P = 0.003,6; 3.78% ± 2.18% vs. 0, P = 0.008,1). TLR4, nuclear factor kappa-B p65, and tumor necrosis factor-α expression in the aorta tissue of the AS group were upregulated, whereas occludin and zonula occludens-1 expression in colon tissues was downregulated. Additionally, metabolomics identified significant differences in 21 metabolites in the feces of the AS group compared to the NC group, with further analysis linking these differences to amino acid metabolism. Conclusions::The Tibetan miniature pig model of early AS induced by high-fat intake displayed pronounced chronic inflammation. Preliminary findings suggest that the underlying mechanisms may be associated with the LPS/TLR4 pathway and intestinal metabolic disorders.

Objective:To understand the prevalence and characteristics of Rhinovirus (HRV) infection in influenza-like Illness (ILIs) patients in Mianyang, Sichuan province, China.Methods:Throat swabs were collected from patients of ILIs in sentinel hospitals in Mianyang during 2021—2022. Real-time fluorescence quantitative PCR was used to detect 16 common pathogens. The VP4/VP2 coding region genes of HRV positive samples were amplified by nest PCR. The phylogeny, consistency and amino acid variation of different serotypes were analyzed and compared with reference sequences from GenBank database.Results:A total of 332 ILIs′ samples were collected with a virus detection rate of 58.73% (195/332) in Mianyang. Among them, 23 samples (23/332) were HRV-positive, and 18 VP4/VP2 sequences of HRV strains were successfully amplified. It was found that 13 HRV serotypes were detected in ILIs samples in Mianyang, which belonged to three genotypes, namely HRV-A (12 strains), HRV-B (5 strains) and HRV-C (1 strain).Conclusions:HRV was one of the pathogens of ILIs cases in Mianyang during 2021—2022, with HRV-A types as the dominant strains.

【Objective】 To explore the application effect of traditional teaching combined with problem based learning (PBL) in the teaching of transfusion medicine taking the chapter of Human Leukocyte Antigen System and Testing in Clinical Blood Transfusion Testing Technique as an example. 【Methods】 Firstly, practical problems in chapter Human Leukocyte Antigen System and Testing were analyzed. Then, in response to the key and difficult points in the teaching of this chapter, examples were given to illustrate the practical application of traditional teaching combined with PBL in the teaching of Clinical Blood Transfusion Testing Technique from the aspects of teaching objectives, teaching processes, course implementation and implementation effects. The teaching effectiveness was evaluated through a questionnaire. 【Results】 Traditional teaching combined with PBL helped students grasp important knowledge and techniques, break down thinking barriers, grasp internal connections and characteristics, simplify the learning process, stimulate interest in learning and enhance team collaboration. Meanwhile, students' exploration and innovation abilities could be further cultivated through extracurricular expansion by teachers, thus achieving ideal teaching effect. The questionnaire showed that over 90% of the students consider that combining traditional teaching with PBL was beneficial for improving teaching effect. 【Conclusion】 In the teaching of transfusion medicine, adopting traditional teaching combined with PBL according to the textbook content can improve the teaching effect, which is beneficial for the cultivation of comprehensive talents.

Objective:To develop a biomimetic nanoparticle probe of hematoporphyrin monomethyl ether (HMME) coated with breast cancer cell membrane, to observe its ability to target homologous breast cancer cells in vitro, and to investigate its effect of enhanced photoacoustic imaging and sonodynamic therapy (SDT) for breast cancer in vitro.Methods:The cell membrane of breast cancer 4T1 was extracted by chemical cleavage and repeated freezing and thawing. Then the HMME-coated polylactic acid-glycolic acid copolymer biomimetic nanoparticle was prepared by double emulsification and extrusion. The basic characteristics of nanoparticles were detected. The target ability of nanoparticles to homologous breast cancer cells and the enhancement of photoacoustic imaging were observed in vitro. Singlet oxygen sensor green (SOSG) was used to verify the reactive oxygen species (ROS) production of nanoparticles, and its SDT effect on breast cancer cells was evaluated by CCK8 cytotoxicity assay.Results:The size of the prepared CHP-NPs was uniform, the morphology was spherical "core-shell structure" , the particle size was (275.23±8.25)nm, and the surface potential was (-18.43±0.45)mV. It was observed that CHP-NPs could target homologous 4T1 cells under laser confocal microscopy. In vitro photoacoustic imaging experiments show that the photoacoustic signal of nanoparticles increases with the increase of its concentration. According to SOSG probe detection, CHP-NPs could produce ROS under ultrasonic irradiation.When CHP-NPs was incubated with 4T1 cells alone and no ultrasonic irradiation was used, the cell survival rate was not significantly affected. When the concentration was 0.6 mg/ml, the cell survival rate was still 95%. After ultrasonic irradiation, CCK8 experiment showed that the CHP-NPs had a significant SDT effect on breast cancer cells.Conclusions:The biomimetic nanomolecular probe of breast cancer cell membrane is successfully prepared. The probe has good ability to target homologous tumor, and can significantly enhance tumor photoacoustic imaging and SDT effect.

【Objective】 To investigate the frequencies of HLA-B alleles in Jiangsu Han population from Chinese Marrow Donor Program (CMDP) Jiangsu Branch in 8 prefecture-level cities, compare and analyze the distribution of common susceptible alleles at HLA-B locus in 8 prefecture-level cities. 【Methods】 The HLA-B locus of 27 249 unrelated volunteers from CMDP Jiangsu Branch were genotyped by polymerase chain reaction–sequence based typing (PCR-SBT) method, and frequencies of HLA-B alleles of 8 prefecture-level cities were calculated and compared. 【Results】 A total of 145 HLA-B alleles were detected in Jiangsu Han population, among them 104, 113, 83, 88, 88, 85, 84, 72 HLA-B alleles were detected in Changzhou, Nanjing, Zhenjiang, Yangzhou, Huai’an, Suqian, Xuzhou, and Yancheng Han population, respectively. In general, the common alleles in Changzhou, Yangzhou and Zhenjiang were very similar, and the most common alleles were B^*46∶01, 13∶02, 40∶01 and 58∶01. The common alleles in Nanjing, Huai’an and Suqian were rather similar, and the most common alleles were B^*13∶02, 46∶01, 40∶01, 58∶01 and 51∶01. The common alleles in Yancheng were rather similar to Nanjing, Huai’an and Suqian, with only difference in the order of B^*40∶01 and B^*58∶01. The common alleles in Xuzhou were close to Nanjing, Huai’an and Suqian too, with only difference in the order of B^*40∶01 and B^*46∶01. 【Conclusion】 The results reflected the distribution of HLA-B alleles of Jiangsu Han population from 8 prefecture-level cities and the gene distribution was basically consistent with the regional distribution. The major difference in HLA-B locus in Jiangsu 8 prefecture-level cities was B ^* 13∶02 and B ^* 46∶01. The frequency of B ^* 13∶02 in Northern Jiangsu (Huai’an, Suqian, Xuzhou, Yancheng) is much higher than that of B ^* 46∶01, and that in Nanjing is slightly higher than the frequency of B ^* 46∶01, which is consistent with that in North China. The frequency of B ^* 46∶01 in Southern Jiangsu (Changzhou), Yangzhou and Zhenjiang is higher than that of B ^* 13∶02, which is consistent with that in South China. The polymorphism analysis of alleles at HLA-B locus in 8 cities in Jiangsu Province provided significant basic data for the study of the correlation between HLA-B locus genes and diseases, as well as the population genetics and anthropology in Jiangsu Han population.

Objective To explore the impact of IL10-592 (rs1800872) single nucleic acid polymorphism (SNP) on the prognosis of HLA matched unrelated hematopoietic stem cell transplantation (HSCT).Methods The polymorphism of IL10-592 in 104 recipient-donor pairs and 100 healthy volunteers was analyzed with sequence based typing (SBT).Results When the genotype of IL1 0-592 in donors and recipients matched,AA/AA genotype had higher incidence of Ⅲ-Ⅳ aGVHD than AC/AC or CC/CC genotype (47.1％,3.7％,0,P=0.002).When the genotype of IL10-592 in donors and recipients mismatched,recipients with AC genotype or donors with AA genotype,there was significant different incidence of Ⅲ-Ⅳ aGVHD among donors or recipients with different genotype (P=0.046,P=0.041).The recipients with AA genotype had higher incidence of Ⅲ-Ⅳ aGVHD than AC or CC genotype (27.8％ vs 10.2％,11.1％;P=0.072),and higher incidence of intestinal aGVHD (22.2％vs 5.1％,11.1％;P=0.040),lower incidence of 2-year overall survival (OS:48.2％ vs 75.1％,85.7％;P=0.002),lower incidence of 2 year disease free survival (DFS:48.5％ vs 66.3％,76.2％;P=0.045).Patients had higher incidence of Ⅲ-Ⅳ aGVHD with donors of AA genotype than with donors of AC or CC genotype (26.5％ vs 8.9％,0;P=0.024),and higher incidence of intestinal aGVHD (20.4％ vs 4.4％,0;P=0.026).In multivariate analysis,the genotype of IL10-592AA in recipients and donors had increased risk of Ⅲ-Ⅳ aGVHD (OR=3.3,P=0.049;OR=3.9,P=0.043).There were no statistical differences on the incidence of cGVHD and relapse.Conclusion In HLA-10/10 matched unrelated HSCT,the presence of IL10-592 AA genotype in recipients and/or donors is an adverse factor for Ⅲ-ⅣaGVHD,worse OS and 2-year DFS.

Objective To analyze the frequencies of HLA-DQA1 alleles and their clinical values in the donor-recipient HLA-A,-B,-C,-DRB1,-DQB1 (10/10) matched hematopoietic stem cell transplantation (HSCT).Methods This study recruited 127 patients who received allogeneic HSCT and 127 unrelated donors.High-resolution (High Res) DNA typing for HLA-DQA1 alleles were performed on the 254 subjects by using sequence specific oligonucleotide probes (SSOP) and high resolution of sequence specific primer(High Res SSP).Results The DQA1 allele genotypes of 36 pairs of donor-recipient were directly identified by using SSOP.The ambiguous DQA1 allele genotypes of the rest 91 pairs were identified by using High Res SSP.Among the 127 pairs of donor-recipient,5 pairs were HLA-DQA1 alleles mismatched,while the others were all matched.No significant differences in the distribution of HLA-DQA1 alleles were observed between the donors and the recipients.Sixteen HLA-DQA1 alleles were detected in the 127 donors,which were DQA1 * 02 ∶ 01 (19.3％),DQA1* 01 ∶ 02(19.3％),DQA1 * 03 ∶ 02/03 (17.0％),DQA1 *01∶03 (9.8％),DQA1*06∶01(9.1％),DQA1*05∶ 01(7.1％),DQA1*05∶05(5.9％),DQA1*03∶01 (4.7％),DQA1*01 ∶04(2.4％),DQA1*01∶05(2.0％),DQA1*01∶01(1.2％),DQA1*05 ∶ 03(0.8％),DQA1 *05 ∶ 08(0.8％),DQA1*04 ∶ 01(0.4％),DQA1*05 ∶ 06(0.4％) from high to low frequency.Moreover,a new allele was detected in the patients.The haplotypes' frequencies and linkage disequilibrium(LD) analysis of HLA-DQA1 and HLA-DQB1 showed that the most common haplotype was DQA1 *02 ∶ 01-DQB1 *02 ∶ 02(16.1％),followed by DQA1 *03 ∶ 02/03-DQB1 *03 ∶ 03 (11.8％)and DQA1 *01 ∶ 03-DQB1 * 06 ∶ 01 (9.1％).Stronger LD were observed between DQA1 * 02 ∶ 01 and DQB1*02 ∶ 02,DQA1 *03 ∶ 02 and DQB1*03 ∶ 03,DQA1 *01 ∶ 03 and DQB1*06 ∶ 01,HLA-DQA1*06∶01 andDQB1*03 ∶ 01,DQA1*05 ∶ 01 and DQB1*02 ∶ 02(P＜0.001).Conclusion There was strong linkage disequilibrium between HLA-DQA1 and HLA-DQB1 genes.The polymorphism of HLA-DQA1 gene was less than that of HLA-DQB1 gene.No more guidance was provided to donor selection in unrelated donor-recipient HLA matched HSCT by adding HLA-DQA1 genotyping,but it might have clinical application values in HSCT with HLA Ⅱ locus mismatched donor and recipient.

Objective To investigate the effect of gender and age on the apparent diffusion coefficient (ADC) of normal adult pancreas.Methods A total of 383 patients with normal pancreas (290 male,93 female,range from 21 to 78 years of age) were enrolled in this study.The subjects were divided into four groups based on different age (≤40 years,41-50 years,51-60 years and ＞60 years) with patient number of 56,108,139 and 80,respectively.Breath-hold single-shot echo-planar DWI (b value =0,500 s/mm2) was performed to determine ADCs on all patients.The average ADCs was calculated by four ADCs measured from the head to tail part of the pancreas in each patient.Patients with different age or gender were analyzed by independent-samples t test.Effect of gender on ADCs was analyzed using the Mann-Whitney U test.Relationship between ADCs and age was analyzed using Spearman rank-order correlation test,and Kruskal-Wallis H test was used to compare the ADCs among 4 age groups.Results The median pancreatic ADC values in female group(n =93) [1.60 × 10-3 mm2/s (1.47 × 10-3-1.77 × 10-3) mm2/s] was higher than that in male group (n =290) [1.57 × 10-3mm2/s(1.41 × 10-3-1.74 × 10-3)mm2/s].Mann-Whitney U test results showed the mean ADCs was similar between the two groups (Z =1.335,P =0.182).The age distribution was similar between the male [(52 ± 10) years of age] and female [(51 ± 11) years of age]groups (t =0.267,P=0.790).The age spectrum showed that there was no correlation between the average ADC values and age (r =0.016,P =0.752).The median ADC values of the four age groups were 1.58 ×10-3,1.54 × 10-3,1.59 × 10-3 and 1.57 × 10-3 mm2/s,respectively.Kruskal-Wallis H test showed no significant difference of mean ADCs among the age groups (x2 =2.15,P =0.542).Conclusion There is no correlation of ADCs between age and gender in normal adult pancreas.

Objective To investigate the frequencies of human leukocyte antigen ( HLA)-DRB1 alleles of Chinese Marrow Donor Program (CMDP) donors in seven markets of Jiangsu Han population. Methods 2787 donors from CMDP Jiangsu Branch were typed in HLA-DRB1 locus using polymerase chain reaction-sequence-based typing (PCR-SBT) method or polymerase chain reaction-reverse sequence specific oligonucleotide probes (PCR-rSSOP) method. Frequencies of HLA-DRB1 were separately calculated in seven markets and a comparison was made among them. Results A total of 38,33,26,36,37,31,26 alleles in DRB1 locus were separately found in Huai-an, Xuzhou, Taizhou, Suqian, Nanjing, Changzhou, Yangzhou population. The most frequent HLA-DRB1 alleles in Huai-an, Xuzhou, Suqian respectively were DRB1 * 07:01, 09:01, 15:01,12:02; In Nanjing and were DRB1 * 09:01,07:01, 15:01, 12:02; In Yangzhou were DRB1 * 09:01,07:01,15:01,08:03; In Taizhou were DRB1 *09:01, 07:01, 04:05, 15:01; In Changzhou were DRB1 *09:01,07:01, 12:02, 08:03. Conclusion The above results reflected the distribution of HLA-DRB1 alleles in seven markets of Jiangsu Han population, which held abundant polymorphism, and a certain uniqueness in every market. The HLA-DRB1 allele distribution of Nanjing, Yangzhou were the closest groups to northern China, Huaian, Xuzhou, Suqian, Changzhou, Taizhou were close to northern China except DRB1 * 09:01,07:01 in different frequencies. These findings could guide the selection of future donor in CMDP Jiangsu Branch. It also could serve as foundation for HLA related disease research, studies of population genetics and anthropology in Chinese Han population.