Objective To explore the feasibility and reference value of allogeneic lung transplantation and postoperative monitoring in miniature pigs for lung transplantation research. Methods Two miniature pigs (R1 and R2) underwent left lung allogeneic transplantation. Complement-dependent cytotoxicity tests and blood cross-matching were performed before surgery. The main operative times and partial pressure of arterial oxygen (PaO₂) after opening the pulmonary artery were recorded during surgery. Postoperatively, routine blood tests, biochemical blood indicators and inflammatory factors were detected, and pathological examinations of multiple organs were conducted. Results The complement-dependent cytotoxicity test showed that the survival rate of lymphocytes between donors and recipients was 42.5%-47.3%, and no agglutination reaction occurred in the cross-matching. The first warm ischemia times of D1 and D2 were 17 min and 10 min, respectively, and the cold ischemia times were 246 min and 216 min, respectively. Ultimately, R1 and R2 survived for 1.5 h and 104 h, respectively. Postoperatively, in R1, albumin (ALB) and globulin (GLB) decreased, and alanine aminotransferase increased; in R2, ALB, GLB and aspartate aminotransferase all increased. Urea nitrogen and serum creatinine increased in both recipients. Pathological results showed that in R1, the transplanted lung had partial consolidation with inflammatory cell infiltration, and multiple organs were congested and damaged. In R2, the transplanted lung had severe necrosis with fibrosis, and multiple organs had mild to moderate damage. The expression levels of interleukin-1β and interleukin-6 increased in the transplanted lungs. Conclusions The allogeneic lung transplantation model in miniature pigs may systematically evaluate immunological compatibility, intraoperative function and postoperative organ damage. The data obtained may provide technical references for subsequent lung transplantation research.

BACKGROUND:Angiogenesis is the main treatment target of cardiovascular diseases.Bone morphogenetic protein 2 can modulate angiogenesis,but the regulatory effect of endothelial cell-specific bone morphogenetic protein 2 on angiogenesis is unclear. OBJECTIVE:To investigate the effect of endothelial-specific bone morphogenetic protein 2 on angiogenesis. METHODS:(1)Bioinformatics analysis:Cellular expression specificity and abundance of bone morphogenetic protein 2 were meta-analyzed by the PanglaoDB single-cell transcriptome database.The endothelial cell transcriptome sequencing dataset of the mouse hindlimb model and endocardial transcriptome dataset of mice overexpressing bone morphogenetic protein 2 were reanalyzed to evaluate the effect of endothelial cell bone morphogenetic protein 2 on the angiogenesis pathway.(2)Validation in vivo:After establishing the mouse hindlimb model,we compared the blood perfusion between the affected and sham limb at 7,14,and 21 days.The expression of the colocation of bone morphogenetic protein 2 and CD31 was explored by immunofluorescence and immunohistochemical staining.(3)Validation in vitro:The cultured human umbilical vein endothelial cells in vitro were divided into a control group,a hypoxia group,and a bone morphogenetic protein 2 inhibitor Noggin intervention group.After being cultured for 24 hours,the angiogenesis of endothelial cells in each group was observed. RESULTS AND CONCLUSION:(1)Endothelial cells are an important cell subgroup expressing bone morphogenetic protein 2.Both in the mouse hindlimb ischemia model and endocardial cells overexpressing bone morphogenetic protein 2,bone morphogenetic protein 2 was significantly up-regulated,and the angiogenesis pathway was significantly activated.(2)In the mouse hindlimb model,bone morphogenetic protein 2-positive blood vessels around neoangiogenesis increased significantly at 7 days of ischemia(P<0.05),and decreased significantly after 2 weeks of ischemia(P<0.001).(3)In umbilical vein endothelial cells cultured in vitro,after hypoxic intervention,the migration and sprouting of endothelial cells increased significantly,and the expression of angiogenesis factors vascular endothelial growth factor and platelet-derived growth factor was significantly increased.Noggin significantly reduced hypoxia-induced endothelial cell angiogenesis(P<0.001)and down-regulated the expression of vascular endothelial growth factor and platelet-derived growth factor(P<0.01).(4)These findings verify that endothelial cell-specific bone morphogenetic protein 2 can regulate angiogenesis,and targeting endothelial cell bone morphogenetic protein 2 is a promising way to improve angiogenesis.

Objective To explore the construction of α-1,3-galactosyltransferase (GGTA1) gene-knockout (GTKO) Diannan miniature pigs and the kidney xenotransplantation from pigs to rhesus macaques, and to assess the effectiveness of GTKO pigs. Methods The GTKO Diannan miniature pigs were constructed using the CRISPR/Cas9 gene-editing system and somatic cell cloning technology. The phenotype of GTKO pigs was verified through polymerase chain reaction, Sanger sequencing and immunofluorescence staining. Flow cytometry was used to detect antigen-antibody (IgM) binding and complement-dependent cytotoxicity. Kidney xenotransplantation was performed from GTKO pigs to rhesus macaques. The humoral immunity, cellular immunity, coagulation and physiological indicators of the recipient monkeys were monitored. The function and pathological changes of the transplanted kidneys were analyzed using ultrasonography, hematoxylin-eosin staining, immunohistochemical staining and immunofluorescence staining. Results Single-guide RNA (sgRNA) targeting exon 4 of the GGTA1 gene in Diannan miniature pigs was designed. The pGL3-GGTA1-sgRNA1-GFP vector was transfected into fetal fibroblasts of Diannan miniature pigs. After puromycin selection, two cell clones, C59# and C89#, were identified as GGTA1 gene-knockout clones. These clones were expanded to form cell lines, which were used as donor cells for somatic cell nuclear transfer. The reconstructed embryos were transferred into the oviducts of trihybrid surrogate sows, resulting in 13 fetal pigs. Among them, fetuses F04 and F11 exhibited biallelic mutations in the GGTA1 gene, and F04 had a normal karyotype. Using this GTKO fetal pig for recloning and transferring the reconstructed embryos into the oviducts of trihybrid surrogate sows, seven surviving piglets were obtained, all of which did not express α-Gal epitope. The binding of IgM from the serum of rhesus monkey 20# to GTKO pig PBMC was reduced, and the survival rate of GTKO pig PBMC in the complement-dependent cytotoxicity assay was higher than that of wild-type pig. GTKO pig kidneys were harvested and perfused until completely white. After the left kidney of the recipient monkey was removed, the pig kidney was heterotopically transplanted. Following vascular anastomosis and blood flow restoration, the pig kidney rapidly turned pink without hyperacute rejection (HAR). Urine appeared in the ureter 6 minutes later, indicating successful kidney transplantation. The right kidney of the recipient was then removed. Seven days after transplantation, the transplanted kidney had good blood flow, the recipient monkey's serum creatinine level was stable, and serum potassium and cystatin C levels were effectively controlled, although they increased 10 days after transplantation. Seven days after transplantation, the levels of white blood cells, lymphocytes, monocytes and eosinophils in the recipient monkey increased, while platelet count and fibrinogen levels decreased. The activated partial thromboplastin time, thrombin time and prothrombin time remained relatively stable but later showed an upward trend. The recipient monkey survived for 10 days. At autopsy, the transplanted kidney was found to be congested, swollen and necrotic, with a small amount of IgG deposition in the renal tissue, and a large amount of IgM, complement C3c and C4d deposition, as well as CD68⁺ macrophage infiltration. Conclusions The kidneys of GTKO Diannan miniature pigs may maintain normal renal function for a certain period in rhesus macaques and effectively overcome HAR, confirming the effectiveness of GTKO pigs for xenotransplantation.

Alzheimer's disease (AD) is characterized by cognitive and functional deterioration, with pathological features such as amyloid-beta (Aβ) aggregates in the extracellular spaces of parenchymal neurons and intracellular neurofibrillary tangles formed by the hyperphosphorylation of tau protein. Despite a thorough investigation, current treatments targeting the reduction of Aβ production, promotion of its clearance, and inhibition of tau protein phosphorylation and aggregation have not met clinical expectations, posing a substantial obstacle in the development of drugs for AD. Recently, artificial intelligence (AI), computational biology (CB), and systems biology (SB) have emerged as promising methodologies in AD research. Their capacity to analyze extensive and varied datasets facilitates the identification of intricate patterns, thereby enriching our comprehension of AD pathology. This paper provides a comprehensive examination of the utilization of AI, CB, and SB in the diagnosis of AD, including the use of imaging omics for early detection, drug discovery methods such as lecanemab, and complementary therapies like phototherapy. This review offers novel perspectives and potential avenues for further research in the realm of translational AD studies.

Lipid metabolism,as the basis of life maintenance,is a prerequisite for cell survival,and lipid homeostasis can rapidly respond to metabolic changes in a coordinated manner.In cancers,there is an increase in lipid metabolism in cancer cells to meet the requirements for plasma membrane synthesis and energy production.Abnormal lipid metabolism plays an important role in the progression of primary liver cancer.This article reviews the association between abnormal lipid metabolism and primary liver cancer,in order to find targets for the prevention and treatment of primary liver cancer.

Objective To analyze the composition of microbial community in gut of elderly male Nycticebus bengalensis,aiming to explore the health impact factors associated with artificial captivity.Methods Fecal samples of 4 adult male Nycticebus bengalensis were collected and the V4 region was amplified using bacterial 16S rDNA universal primers.Illumina NovaSeq sequencing platform was used for microbial sequencing analysis.The complexity and similarity of the samples were analyzed using the QIIME software analysis tool.The species structure and abundance of the intestinal flora were analyzed at the phylum and genus levels based on validated data.The PICRUSt software was applied to predict the metabolic function of the flora.Results The results showed that the diversity index(Shannon index)of the flora in the youngest Nycticebus bengalensis was higher than that of the others.PCoA analysis showed that the bacterial community compositions of the four samples had a certain degree of similarity.Bacteria identified in the Nycticebus bengalensis feces included 12 phyla,18 classes,28 orders,49 families,93 genera,and 59 species.Among them,the dominant phyla were Bacteroidetes and Firmicutes with average relative abundances of 46％and 28％,respectively;The genera Bacteroides,Bifidobacterium,and Fusobacterium had higher abundances,with relative abundances of 33％,6％,and 6％,respectively;The beneficial genus Bifidobacterium was found in all samples,with the highest relative abundance found in the younger Nycticebus bengalensis.PICRUSt function prediction analysis showed that the abundance of the functional genes related to amino acid transport and metabolism,carbohydrate transport and metabolism,was relatively high.Conclusion The use of Illumina NovaSeq high-throughput sequencing technology can comprehensively detect the fecal microbial community of the Nycticebus bengalensis.The bacterial composition of Nycticebus bengalensis has rich diversity.Many bacteria are not identified and have higher relative abundance,which need further study.

Objective:To explore the genetic characteristics of a fetus with sex chromosome abnormality indicated by non-invasive prenatal testing (NIPT) at 25 + gestational weeks. Methods:A pregnant woman who was admitted to the Taizhou Hospital for abnormal NIPT result on January 6, 2023 was selected as the study subject. Relevant clinical data was collected. The fetus was subjected to chromosomal karyotyping analysis, copy number variation sequencing (CNV-seq), fluorescence in situ hybridization (FISH), and multiplex PCR assays. Results:NIPT had suggested monosomy of X chromosome. The fetus was found to have a chromosomal karyotype of 45, X[59]/46, X, del(Y)(q11.2)[17] at 30 + weeks of gestational age. CNV-seq suggested the presence a 7.98 Mb deletion at Yq11.222q12 and a mosaicism 16.92 Mb deletion. FISH suggested that the fetus harbored two SRY genes and a mosaicism sex chromosomal abnormality, and multiplex PCR revealed that its AZF b+ c region was completely deleted. C-banded karyotyping showed darkly stained dense mitotic granules at both ends of the Y chromosome. The fetus was ultimately determined as a 45, X/46, X, idic(Y)(q11.2) mosaicism. Following elected abortion, testing of the fetal tissue confirmed the presence of 45, X/46, XY mosaicism, and CNV-seq result of the placental tissue was compatible with that of NIPT. CNV-seq analysis of the couple revealed no obvious abnormality. Conclusion:With combined NIPT, karyotyping, CNV-seq, FISH and multiplex PCR assays, the fetus was diagnosed as a 45, X/46, X, idic(Y)(q11.2) mosaicism with deletion of the AZF b+ c region. Above finding has enabled prenatal diagnosis for the fetus.

Objective:To explore the expression level and clinical significance of serum Hsa_circ_0089761 in patients with cervical cancer.Methods:A total of 107 cervical cancer patients, 80 cervical intraepithelial neoplasia (CIN) patients, and 60 normal control group were selected and analyzed from January 2021 to March 2023 at the Ninth Affiliated People's Hospital of Shanghai Jiao Tong University School of Medicine. We compared the levels of serum Hsa_circ_0089761, squamous cell carcinoma antigen (SCCA), and carcinoembryonic antigen (CEA) among different groups, and analyzed the relationship between the expression level of serum Hsa_circ_0089761 and the clinical and pathological characteristics of cervical cancer. The receiver operating characteristic (ROC) curve was drawn to analyze the diagnostic value of serum Hsa_circ_0089761, SCCA, and CEA levels for cervical cancer. Pearson correlation analysis was used to evaluate the correlation between serum Hsa_circ_0089761 expression levels and SCCA and CEA in cervical cancer patients.Results:The expression levels of serum Hsa_circ_0089761[(2.96±0.95) vs (1.83±0.74), (0.92±0.41)], SCCA[(9.63±1.84)ng/ml vs (2.28±0.65)ng/ml, (1.30±0.27)ng/ml], and CEA[(6.47±2.20)ng/ml vs (1.61±0.57)ng/ml, (1.15±0.12)ng/ml] in the cervical cancer group were significantly higher than those in the CIN group and the control group (all P<0.001), and the serum Hsa_circ_0089761 expression levels in the CIN group were significantly higher than those in the control group ( P<0.001). Cervical cancer patients in stages Ⅲ-Ⅳ, with low differentiation, lymph node metastasis, infiltration depth ≥1/2 of the muscle layer, positive SCCA, and positive CEA had significantly higher levels of serum Hsa_circ_0089761 expression (all P<0.05). The ROC curve analysis showed that the specificity of diagnosing cervical cancer was highest (85.00%) for Hsa_circ_0089761 ≥2.25, and the area under the ROC curve (AUC) for diagnosing cervical cancer in combination with SCCA was highest (0.932, 95% CI: 0.874-0.993), with the highest accuracy (89.30%). The sensitivity of the combination of Hsa_circ_0089761+ SCCA+ CEA in diagnosing cervical cancer was highest (96.26%). The correlation analysis results showed that the serum Hsa_circ_0089761 expression levels in cervical cancer patients were positively correlated with SCCA ( r=0.775, P<0.001) and CEA ( r=0.613, P<0.001). Conclusions:The expression level of serum Hsa_circ_0089761 in cervical cancer patients is significantly increased, which is related to clinical and pathological characteristics. The combination of Hsa_circ_0089761 and SCCA detection has high value in the diagnosis of cervical cancer.

Objective:To investigate the effects of peritoneal dialysis combined with hemodialysis on improving cardiac and renal function, peritoneal function, and nutritional status, and reducing complications in patients with end-stage renal disease.Methods:A total of 90 patients with end-stage renal disease who received treatment at Lishui Municipal Central Hospital and Lishui People's Hospital from January 2019 to May 2022 were retrospectively included in this study. The patients were divided into a control group and an observation group, with 45 patients in each group, based on the treatment methods used. The control group received peritoneal dialysis treatment, while the observation group received a combination of peritoneal dialysis and hemodialysis treatment. The changes in cardiac and renal function, peritoneal function, nutritional status, and the occurrence of complications were observed in both groups.Results:After treatment, the observation group showed significantly lower values in the cardiothoracic ratio [(57.02 ± 3.35)%], brain natriuretic peptide [(8 849.34 ± 124.65) ng/L], left ventricular mass index [(181.32 ± 8.56) g/m2], blood urea nitrogen [(14.04 ± 1.94) mmol/L], and serum creatinine [(181.47 ± 27.06) μmol/L] compared with the control group [(58.92 ± 3.11)%, (15 126.39 ± 322.82) ng/L, (187.28 ± 8.95) g/m2, (18.49 ± 2.82) mmol/L, (196.56 ± 31.07) μmol/L, t = 2.79, 121.68, 3.23, 8.72, 2.46, all P < 0.05]. Additionally, the observation group had significantly higher levels of ejection fraction [(63.47 ± 5.23)%], albumin [(52.98 ± 6.37) g/L], and hemoglobin [(114.94 ± 13.61) g/L] compared with the control group [(60.46 ± 5.18)%, (47.01 ± 6.04) g/L, (98.04 ± 10.93) g/L, t = -2.74, -4.52, -6.49, all P < 0.05]. The observation group also exhibited a significantly lower dialysate/plasma creatinine ratio [(0.56 ± 0.09)] and a lower level of β 2-microglobulin [(18 032.29 ± 718.11) mg/L] compared with the control group [(0.61 ± 0.07), (18 424.42 ± 736.43) mg/L, t = 2.54, 2.06, both P < 0.05]. The incidence of complications in the observation group was 11.11% (5/45), which was significantly lower than that in the control group [31.11% (14/45), χ2 = 4.27, P < 0.05]. Conclusion:Peritoneal dialysis combined with hemodialysis can improve cardiac and renal function, peritoneal function, and nutritional status and decrease the incidence of complications in patients with end-stage renal disease. This approach deserves to be clinically promoted. The findings of this study are of significant innovation and scientific value.

Porcine circovirus type 2(PCV2)mainly damages the immune cells of pigs,causing lym-phocyte depletion and immune suppression.Interleukin(IL)-15 regulates immune functions wide-ly,and plays an important regulatory role in the survival,proliferation,differentiation and immune function of a variety of immune cells such as natural killer(NK),CD8+T cells and NKT cells.In this study,in order to determine the effect of PCV2 on IL-15 expression,4-week-old piglets(n=4)were infected with PCV2 and the negative control group(n=4)was set up.On day 7 post-infec-tion,the inguinal lymph nodes of the infected and control groups were collected,and porcine cyto-kine antibody microarray(QAP-CYT-1)was employed to quantify the expression of cytokines in the tissues,screen for differential cytokines,and GO and KEGG enrichment analysis for IL-15 were conducted.Real-time quantitative fluorescent PCR(qPCR)and ELISA were used to verify the level of IL-15 mRNA and protein,and those in porcine peripheral blood mononuclear cells(PBMC)and serum were simultaneously detected.Compared with the negative control group,the expression lev-el of IL-15 was significantly reduced in the infected group(P＜0.05);IL-15 was mainly involved in cytokine-cytokine receptor interactions,immune responses,cellular activation,and the regulation of JAK-STAT and TNF signaling pathways.The levels of IL-15 mRNA and protein in inguinal lymph nodes in the infected group were significantly lower than those in the control group(P＜0.05),which was consistent with the detection results of QAP-CYT-1.However,there was no significant difference in IL-15 mRNA and protein levels in PBMC and serum.These results indicate that PCV2 can inhibit IL-15 in the inguinal lymph node microenvironment of piglets.This study can provide important information for further revealing the immunosuppressive mechanism of PCV2.