Objective: To design and establish a new method for flow cytometry-based detection of commonly observed highly expressed antigens on red blood cells, and to further evaluate the differences and distribution characteristics of antigen expression levels between ABO blood type homozygotes and heterozygotes in healthy individuals. Methods: Residual blood samples after donor blood type identification by Shanghai Blood Center in April 2024 were collected. Among them, samples of 19 homozygous and 19 heterozygous individuals of type A and type B were selected. Then the expression level of ABO antigen on red blood cells were detected using the new method established in this study and the traditional aldehyde fixed red blood cell method. Both methods were tested independently three times and the results were compared. Results: The mean values of the three detection results of the new method was (×10 /RBC): AA homozygous 3.3±0.5, AO heterozygous 2.8±0.3, BB homozygous 3.6±0.3, BO heterozygous 3.1±2.8. The mean values of the three detection results of the aldehyde fixation method were AA homozygous 5.9±0.9, AO heterozygous 5.0±1.4, BB homozygous 3.8±0.6, and BO heterozygous 3.3±0.4. The average antigen distribution of each genotype followed a normal distribution. Comparing the average antigen expression levels of homozygotes and heterozygotes, both methods showed that A/B homozygotes had higher antigen levels than heterozygotes, with AA being 1.17 to 1.18 times that of AO and BB being 1.15 to 1.16 times that of BO. Comparing the inter batch differences in the three test results of two methods, the new method showed no significant difference in the three test results for four genotypes (P>0.05). The aldehyde fixation method showed significant differences in the test results for all three genotypes (P<0.01) except for BB homozygotes (P>0.05). The reliability and reproducibility of the new method were better than those of the traditional aldehyde fixation method. Conclusion: The antigen expression level of ABO homozygotes is higher than that of heterozygotes, and the difference in antigen level between type A homozygotes and heterozygotes is slightly higher than that of type B. The new method is superior to traditional aldolization fixation methods.

Psoriasis is an incurable chronic inflammatory disease that requires new interventions. Here, we found that fibroblasts exacerbate psoriasis progression by promoting macrophage recruitment via CCL2 secretion by single-cell multi-omics analysis. The natural small molecule celastrol was screened to interfere with the secretion of CCL2 by fibroblasts and improve the psoriasis-like symptoms in both murine and cynomolgus monkey models. Mechanistically, celastrol directly bound to the low-density lipoprotein receptor-related protein 1 (LRP1) β-chain and abolished its binding to the transcription factor c-Jun in the nucleus, which in turn inhibited CCL2 production by skin fibroblasts, blocked fibroblast-macrophage crosstalk, and ameliorated psoriasis progression. Notably, fibroblast-specific LRP1 knockout mice exhibited a significant reduction in psoriasis like inflammation. Taken together, from clinical samples and combined with various mouse models, we revealed the pathogenesis of psoriasis from the perspective of fibroblast-macrophage crosstalk, and provided a foundation for LRP1 as a novel potential target for psoriasis treatment.

Objective:To explore the clinical application value of a deep learning algorithm-based ultrasound biomicroscopy (UBM) image analysis system for primary angles-closure glaucoma (PACG) anterior located ciliary body.Methods:A diagnostic test study was conducted.A total of 2 132 UBM images from 726 eyes of 378 PACG patients who underwent UBM examination were collected at Renmin Hospital of Wuhan University from August 2022 to December 2023.The dataset was divided into a training set of 1 599 images and a test set of 533 images, and a deep learning algorithm was employed to construct a model.An additional 334 UBM images from 101 eyes of 69 PACG patients treated at Huangshi Aier Eye Hospital were selected to conduct external testing.A separate set of another 110 UBM images were selected for a human-machine competition to compare the accuracy and speed between anterior located ciliary body evaluation system and three senior ophthalmologists.Furthermore, eight junior ophthalmologists assessed the 110 UBM images independently without and with the assistance of the model, and the differences between the two evaluations were analyzed to assess the assisstance effect of the model.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Renmin Hospital of Wuhan University (No.WDRY-2022-K109).Results:The model achieved an accuracy of 93.43% for anterior located ciliary body identification in the internal test set, with a sensitivity of 84.30% and a specificity of 97.78%.The model also performed well on the external test set with an accuracy of 92.81%.In the human-machine competition, the model's accuracy was comparable to that of the senior ophthalmologists and outperformed two of the three senior ophthalmologists.The average total time of the three senior ophthalmologists was 726.73 seconds, approximately 12.47 times longer than the model's 58.30 seconds.With model assistance, the diagnostic accuracy of the eight junior ophthalmologists was 86.71%, which was significantly higher than 76.25% without model assistance ( χ2=-7.550, P<0.001).And the image evaluation time was (714.91±213.82)seconds, which was significantly lower than (987.90±238.56)seconds without model assistance ( t=2.774, P<0.05). Conclusions:The UBM image analysis system based on a deep learning algorithm demonstrates high accuracy in diagnosing anterior located ciliary body in PACG and provides a strong support for the UBM image recognition training of junior ophthalmologists.

As a practical carrier for promoting the tiered diagnosis and treatment model, the medical consor-tium is of great significance for balancing medical resources and boosting medical service efficiency. The construction of medical consortiums not only improves the accessibility of high-quality medical resources for patients, but also enhances the diagnostic and treatment level of member units. Meanwhile, it provides space for the leading hospital to adjust the structure of diseases and improve the level of discipline construction. As the core of medical insurance payment reform, DRG, through indicators such as the case mix index(CMI) and the number of diagnosis related group (DRG), provides objective and quantified data support for case management and disease structure optimization, thus effectively guiding the rational allocation of medical resources and the adjustmentof diseases and surgical types within the medical consortium. Comprehensive use of DRG evaluation indicators can construct a multidimensional medical consortium construction evaluation system, provides a clear direction for medical consortium cooperation, thereby promoting the overall healthy and sustainable development of medical consortiums and achieving a win-win situation for all parties involved. This paper, based on the "1+5+1" medical consortium cooperation model centered around Peking Union Medical College Hospital, utilizes DRG indicators to analyze the benefits for patients, member hospitals, and the leading hospital during the medical consortium construction process, with the hope of providing reference for the construction of a medical consortium evaluation system.

OBJECTIVE To investigate the changes in high performance liquid chromatography （HPLC） fingerprint spectra and nucleoside components between Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine and its processed product Succus bambusae pinella preparata， providing a reference for the quality evaluation of the latter. METHODS HPLC fingerprint was established for 10 batches of Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine and its processed product Succus bambusae pinella preparata following the Similarity Evaluation System of TCM Chromatographic Fingerprints （2012 Edition）. Hierarchical cluster analysis （HCA）， principal component analysis （PCA）， and orthogonal partial least squares-discriminant analysis （OPLS- DA） were conducted on their common peaks. The contents of four nucleoside components， hypoxanthine， uridine， adenine， and guanosine， in both Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine and Succus bambusae pinella preparata were determined. RESULTS The similarity between the fingerprints of the 10 batches of Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine， Succus bambusae pinella preparata， and their corresponding reference fingerprints ranged from 0.851 to 0.990. A total of 10 common peaks were obtained for both samples， and 4 components were identified as hypoxanthine， uridine， adenine， and guanosine. The results of HCA， PCA and OPLS-DA showed that the samples of Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine and Succus bambusae pinella preparata were clustered into separate categories， with OPLS-DA selecting 4 differential components between them， ranked by variable importance projection values as peak 8， peak 1， peak 6 （adenine） and peak 10. The content determination results showed that the average contents of hypoxanthine， uridine， adenine and guanosine in Succus bambusae pinella preparata declined by 15.90%， 12.00%， 26.04% and 22.18% compared to Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine， respectively， with statistically significant differences in the contents of hypoxanthine， adenine and guanosine （P＜0.05 or P＜0.01）. CONCLUSIONS The established fingerprint and content determination methods are simple to operate and have good repeatability， which are suitable for qualitative and quantitative analysis of Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine and Succus bambusae pinella preparata. The average contents of the four nucleoside components decreased after the processing of Succus bambusae pinella preparata.

To promote the ideological and political construction of the doctor-patient communication course, the research group discussed the subject characteristics and proposed the goals, principles, elements, and paths of the ideological and political construction of the doctor-patient communication course combined with practical teaching and relevant policy documents. Besides, this paper put forward the top-level framework design for the implementation, curriculum assessment, and evaluation indicators of the ideological and political construction, and developed the Guidelines for Ideological and Political Teaching of the Doctor-patient Communication Course and related teaching evaluation indicators, with a view to providing reference evaluation standards for the ideological and political construction of the doctor-patient communication course in China.

ObjectiveTo investigate the synergistic effect of Coptidis Rhizoma crude polysaccharide （CCP） and berberine （BBR） in treating ulcerative colitis （UC） model mice. MethodThirty male BALB/c mice were randomized into five groups. Except the 6 mice in the normal group， the rest were given 5% dextran sodium sulfate in their daily drinking water to establish the UC model. After modeling， the mice were administrated with corresponding agents by gavage once daily for 4 days： BBR （100 mg·kg^-1） group， BBR （100 mg·kg^-1） + low-dose （22.8 mg·kg^-1） CCP group， BBR （100 mg·kg^-1） + high-dose （45.6 mg·kg^-1） CCP group. The mice in the model group and normal group were administrated with the same volume of normal saline. At the end of the experiment， the mice were sacrificed for the collection of colon， and the expression of tight junction proteins zonula occluden-1 （ZO-1）， Claudin-1， and Occludin in colon tissue was detected by Western blot. With the normal group as the control， the disease activity index （DAI） score， colon length， colon histomorphology， and expression levels of tight junction proteins in other groups were evaluated. ResultCompared with the normal group， the modeling down-regulated the protein levels of ZO-1， Claudin-1， and Occludin （P<0.01）. Compared with the model group， BBR did not significantly change the protein level of Claudin-1 and up-regulated those of ZO-1 and occludin （P<0.01）. The expression levels of Claudin-1， ZO-1， and Occludin were up-regulated in BBR + CCP groups （P<0.01）. The expression levels of tight junction proteins in BBR + CCP groups were significantly higher than those in the BBR group （P<0.05）. ConclusionThe administration of CCP combined with BBR can effectively ameliorate intestinal mucosal barrier damage in the mice with UC.

ObjectiveTo determine the pathogens and drug resistance in patients with bacterial diarrhea in Lishui City of Zhejiang Province from 2015 to 2019， and provide scientific evidence for prevention and treatment of bacterial diarrhea. MethodFecal specimens were collected from patients with diarrhea in the People’s Hospital of Lishui City from 2015 to 2019. Bacteria were identified by time-of-flight mass spectrometer and serum agglutination reaction. Drug sensitivity in the suspected bacteria was identified by VITEK 2 Compact system. ResultsA total of 2 937 fecal samples were tested from 2015 to 2019， of which 191 were positive for bacteria. The prevalence was 6.65% in male and 6.32% in female. It was highest in the age group 21‒30 years old， followed by the group 51‒60 years old. Summer was the season with the highest prevalence of bacteria. Furthermore，the bacterial species included salmonella （3.98%）， Vibrio parahaemolyticus （1.43%）， aeromonas （0.48%）， shigella （0.37%） and other bacteria （3.66%）. Salmonella had high resistance to cefuroxime and amikacin. Vibrio parahaemolyticus and shigella had high resistance to ampicillin. Aeromonas had high resistance to ampicillin and ampicillin/sulbactam. ConclusionPrevalence of bacteria differs by gender， age and seasons in patients with bacterial diarrhea in Lishui from 2015 to 2019. Salmonella is the principal pathogen in bacterial diarrhea. Additionally， multiple drug resistance is commonly identified. Therefore， it warrants strengthening the pathogenic surveillance on bacteria and drug resistance in bacterial diarrhea.

【Objective】 To construct an in-vitro model of erythrocyte antibody-mediated complement activation, and establish quantitative detection methods based on flow cytometry and spectrophotometry, so as to explore the correlation of anti-body titers and complement activation speed, and provide a methodological basis for studying the adverse transfusion reactions of anti-body mediated complement hemolysis. 【Methods】 Mouse monoclonal antibody that recognized human C3b and fluorescent secondary antibody were used to label C3b fragments on erythrocytes, and the deposition of C3b fragments after complement activation was detected by flow cytometry. The absorbance at 540 nm of the supernatant in the complement activation reaction system was measured by spectrophotometry as the amount of hemoglobin released was related to the absorbance. 【Results】 The complement activation system was constructed according to the ratio of 3% red blood cell suspension (mixed for 6 people) 1∶anti-Tj^a 1∶complement 2. The repeatability was good (P value>0.05) as different red blood cell mixtures had been used to repeat the detection reaction system. When using 32×, 64× and 128× dilutions of anti-Tj^a mediated complement activation, the deposition of C3b fragments has been detected by flow cytometry at 30 s, 1 min and 2 min, respectively, and MFI peaked at 5 min, 10 min and 30 min, respectively. No obvious hemolysis has been observed within 1.5 h. 【Conclusion】 In vitro model of anti-Tj^a-mediated complement activation demonstrates the speed of complement activation is related to the concentration of antibody. At a certain antibody concentration, the speed of complement activation has been slowed down, and no obvious hemolysis observed.

Objective:To investigate the causes of strong pathogenicity of Bacillus cereus ( B. cereus) in a mouse model of B. cereus endophthalmitis and the factors that might be related to the prognosis of the disease. Methods:C57BL/6J mice aged 6-8 weeks were injected with 1 μl PBS solution containing 100 CFU B. cereus into the vitreous cavity to construct traumatic endophthalmitis model, and a control group was set up by injecting the contralateral eyeball with 1 μl sterile PBS. A mouse model of Staphylococcus epidermidis ( S. epidermidis) endophthalmitis was constructed in the same way as disease control group. Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA) were used to detect the levels of inflammatory cytokines at different time points. Histology, electroretinogram and transmission electron microscopy were used to detect the progression of endophthalmitis and retinal function at different time points. Results:B. cereus grew significantly faster than S. epidermidis in the eyes of C57BL/6 mice and gradually moved to the cornea 12 h after infection. The results of transmission electron microscopy showed that many more B. cereus were found in the iris with sparse pigment particles, while S. epidermidis could not be detected in the anterior segment after infection. The electroretinogram results showed that the amplitude of A wave and B wave of mice with B. cereus endophthalmitis decreased significantly 6 h after infection, and the B wave could not be detected 12 h after infection. Moreover, the amplitude reduction at different time points was significantly larger than that in the S. epidermidis endophthalmitis group. Histological examination found that compared with the S. epidermidis endophthalmitis group, the mice with B. cereus endophthalmitis had significantly increased inflammatory cells in the anterior chamber and vitreous cavity with a higher degree of infiltration, which was more destructive to the tissue structure. ELISA results showed that the activity of myeloperoxidase (MPO) was significantly stronger in the B. cereus endophthalmitis group than in the S. epidermidis endophthalmitis group, suggesting that a much more severe inflammation was induced. The expression of IL-6, TNF-α and IL-1β at the transcription and protein levels in the mouse model of B. cereus endophthalmitis were significantly higher than those in the mice with S. epidermidis endophthalmitis. Conclusions:B. cereus could induce severe endophthalmitis and tissue destruction in the eye due to its rapid growth and migration ability, which was an important factor leading to vision loss.