BACKGROUND:Nobiletin has been found to improve lipopolysaccharide-induced abnormal activation of microglia,excessive release of inflammatory factors and redox imbalance.However,the specific mechanism is not fully understood. OBJECTIVE:To investigate the molecular mechanism by which nobiletin can inhibit lipopolysaccharide-induced inflammation in BV2 microglia. METHODS:Passage 3 BV2 microglia were divided into three groups:control group was cultured for 24 hours(without any treatment).Lipopolysaccharide group was treated with 10 μg/mL lipopolysaccharide for 24 hours.Lipopolysaccharide+nobiletin group was treated with 20 μmol/L nobiletin for 6 hours and then 10 μg/mL lipopolysaccharide for 24 hours.After the processing,cell proliferation was detected by CCK-8 assay.The level of intracellular reactive oxygen species was detected by fluorescent probe.The mRNA expression levels of nuclear factor κB p65,tumor necrosis factor α,and interleukin-1β were detected by qRT-PCR.The protein expression levels of nuclear factor κB p65,p-nuclear factor κB p65,tumor necrosis factor α,and interleukin-1β were detected by western blot assay. RESULTS AND CONCLUSION:(1)Compared with the control group,the proliferation activity of lipopolysaccharide group was decreased(P<0.001).Compared with the lipopolysaccharide group,the cell proliferation activity of lipopolysaccharide+nobiletin group was increased(P<0.001).(2)Compared with the control group,the level of intracellular reactive oxygen species was increased in the lipopolysaccharide group(P<0.001).Compared with the lipopolysaccharide group,the level of intracellular reactive oxygen species was decreased in the lipopolysaccharide+nobiletin group(P<0.01).(3)Compared with the control group,the mRNA expression levels of tumor necrosis factor α and interleukin-1β were increased in the lipopolysaccharide group(P<0.001,P<0.01).Compared with the lipopolysaccharide group,mRNA expression levels of tumor necrosis factor α and interleukin-1β were decreased in the lipopolysaccharide+nobiletin group(P<0.01,P<0.05).(4)Compared with the control group,the protein expression levels of p-nuclear factor κB p65,tumor necrosis factor α,and interleukin-1β in were increased the lipopolysaccharide group(P<0.001).Compared with the lipopolysaccharide group,the expression of p-nuclear factor κB p65,tumor necrosis factor α,and interleukin-1β was decreased in the lipopolysaccharide+nobiletin group(P<0.001).(5)These findings suggest that nobiletin attenuates lipopolysaccharide-induced inflammatory response in BV2 microglia by suppressing nuclear factor-κB signaling pathway.

BACKGROUND: The primary limitation to kidney transplantation is organ shortage. Recent progress in gene editing and immunosuppressive regimens has made xenotransplantation with porcine organs a possibility. However, evidence in pig-to-human xenotransplantation remains scarce, and antibody-mediated rejection (AMR) is a major obstacle to clinical applications of xenotransplantation. METHODS: We conducted a kidney xenotransplantation in a brain-dead human recipient using a porcine kidney with five gene edits (5GE) on March 25, 2024 at Xijing Hospital, China. Clinical-grade immunosuppressive regimens were employed, and the observation period lasted 22 days. We collected and analyzed the xenograft function, ultrasound findings, sequential protocol biopsies, and immune surveillance of the recipient during the observation. RESULTS: The combination of 5GE in the porcine kidney and clinical-grade immunosuppressive regimens prevented hyperacute rejection. The xenograft kidney underwent delayed graft function in the first week, but urine output increased later and the single xenograft kidney maintained electrolyte and pH homeostasis from postoperative day (POD) 12 to 19. We observed AMR at 24 h post-transplantation, due to the presence of pre-existing anti-porcine antibodies and cytotoxicity before transplantation; this AMR persisted throughout the observation period. Plasma exchange and intravenous immunoglobulin treatment mitigated the AMR. We observed activation of latent porcine cytomegalovirus toward the end of the study, which might have contributed to coagulation disorder in the recipient. CONCLUSIONS 5GE and clinical-grade immunosuppressive regimens were sufficient to prevent hyperacute rejection during pig-to-human kidney xenotransplantation. Pre-existing anti-porcine antibodies predisposed the xenograft to AMR. Plasma exchange and intravenous immunoglobulin were safe and effective in the treatment of AMR after kidney xenotransplantation.
Transplantation, Heterologous/methods* ; Kidney Transplantation/methods* ; Heterografts/pathology* ; Immunoglobulins, Intravenous/administration & dosage* ; Graft Survival/immunology* ; Humans ; Animals ; Sus scrofa ; Graft Rejection/prevention & control* ; Kidney/pathology* ; Gene Editing ; Species Specificity ; Immunosuppression Therapy/methods* ; Plasma Exchange ; Brain Death ; Biopsy ; Male ; Aged

Objective: To investigate the characteristics of HIV-1 molecular transmission network among HIV/AIDS patients in Zhoushan City, Zhejing Province. Methods: The newly reported HIV/AIDS cases in Zhoushan City from 2020 to 2022 were selected. Basic information was collected and whole blood samples were obtained at the initial follow-up. The pol gene sequences of HIV-1 were amplified by RT-PCR and nested-PCR. HIV-1 subtypes were identified by Neighbor-Joining phylogenetic trees. The HIV-1 molecular transmission network was built and analyzed using Cytoscape 3.6.1 software. Results: A total of 222 HIV/AIDS cases were reported in Zhoushan City from 2020 to 2022, 200 whole blood samples were collected, and 152 sequences were obtained successfully, including 122 males (80.26%), 75 cases aged 50 years and above (49.34%), 109 cases with a junior high school education or below (71.71%), and 63 cases with commercial heterosexual contact (41.45%). The main subtypes were CRF07_BC and CRF01_AE, accounting for 45.39% and 21.05%, respectively. When the threshold of genetic distance was set to 1%, 20 molecular clusters were formed in 69 cases, with a clustering rate of 45.39%. Using the molecular network constituted by reported HIV/AIDS cases in 2020 as the baseline network, there were 2 active molecular clusters with ≥5 new cases in 2022, each with 9 cases, characterized mainly by individuals aged 50 or above, with a junior high school education or below, and transmission through commercial heterosexual sex. Conclusions The predominant HIV-1 subtypes among HIV/AIDS cases in Zhoushan City are CRF07_BC and CRF01_AE. Transmission through commercial heterosexual contact among middle-aged and elderly people is a main mode of HIV transmission.

Objective:To investigate the clinical value of targeted sequencing panel in the detection of genetic variation in neonates in neonatal intensive care unit (NICU).Methods:All neonates (≤28 d of age) admitted in the NICU (case group) and 200 full-term healthy neonates born with no obvious phenotypic abnormalities of Huzhou Maternity and Child Health Care Hospital were enrolled in this prospective study from November 2022 to January 2023. Based on a list of preventable and treatable rare diseases as well as newly screened diseases in China, a targeted sequencing panel suitable for Chinese newborns was designed to target the pathogenic genes and mutation sites associated with 601 genes and 542 diseases. Dried blood spot specimens were prepared and analyzed by the targeted sequencing panel. Pathogenic sites detected by the panel sequencing were verified using Sanger sequencing. The genetic testing results were analyzed according to the clinical features of the neonates. According to the number of primary clinical diagnosis index (including premature infants, neonatal hyperbilirubinemia, hemorrhagic diseases, neonatal infections, ventricular septal defect/patent ductus arteriosus, and others), these patients were divided into four groups with 1, 2, 3, and ≥4 diagnosis index, respectively. Chi-square test and linear correlation Chi-square test were used for statistical analysis. Results:There were 173 patients in the case group and 30.6% (53/173) of them carried pathogenic variants, including 52 positive for pathogenic genes and one with chromosome copy number variant. The positive rate of pathogenic genes was significantly higher in the case group than in the control group [30.1% (52/173) vs. 15.0% (30/200), χ 2=12.26, P<0.001]. Fourteen pathogenic genes were detected in the case group, including FLG, UGT1A1, G6PD, MYH7, AR, ABCC2, ACADS, CYP21A2, GJB2, MEFV, PAH, PKHD1, SCN4A, and HBA. In the case group, the detection rate of pathogenic variants in jaundiced neonates was higher than that in non-jaundiced neonates [35.2% (44/125) vs. 18.8% (9/48), χ 2=4.42, P=0.036]. However, there were no statistically significant differences in the detection rates of pathogenic variants between male and female infants, infants born to mothers of advanced maternal age or not, infants born to mothers with or without gestational diabetes mellitus, premature and term infants, or infants with or without hemorrhagic disorders, neonatal infections, or ventricular septal defects/patent ductus arteriosus in the case group (all P>0.05). The detection rate of pathogenic variants showed a linear increase in infants with 1, 2, 3, and ≥4 diagnosis index [21.1% (8/38), 25.4% (15/59), 38.2% (13/34), and 40.5% (17/42); linear correlation χ 2=4.84, P=0.028]. In the case group, seven genes with a high detection rate of genetic variation (including positive pathogenic genes and carriers) were UGT1A1 [had the highest detection rate, 24.9% (43/173)], GJB2, FLG, DUOX2, ABCA4, G6PD, and MUT. Seven loci with higher mutation frequency were c.211G>A(p.Gly71Arg), c.1091C>T(p.Pro364Leu), c.-41_-40dupTA, and c.686C>A(p.Pro229Gln) in the UGT1A1 gene, c.109G>A(p.Val37Ile) in the GJB2 gene, and c.12064A>T(p.Lys4022Ter) and c.3321del(p.Gly1109GlufsTer13) in the FLG gene. Conclusion:This panel sequencing can provide effective genetic testing for neonates in NICU, especially in children with complex clinical diagnosis.

Organ shortage has become one of the major challenges hindering the development of organ transplantation. Xenotransplantation is one of the most valuable methods to resolve global organ shortage. In recent years, the development of genetic engineering technique and research and development of new immunosuppressant have provided novel theoretical basis for xenotransplantation. International scholars have successively carried out researches on xenotransplantation in genetically modified pigs to non-human primates or brain death recipients, making certain substantial progresses. However, most of the researches are still in the preclinical stage, far from clinical application. Therefore, according to the latest preclinical experimental research progress at home and abroad, the history of xenotransplantation, the development of gene modification technology, xenotransplantation rejection and immunosuppression regimens were reviewed, aiming to provide reference for subsequent research of xenotransplantation, promote clinical application of xenotransplantation and bring benefits to more patients with end-stage diseases.

Objective To explore the clinical effect and safety of carrellizumab combined with albumin-bound paclitaxel on the treatment of patients with locally advanced esophageal cancer.Methods Ninety-eight patients with locally advanced esophageal cancer were randomly divided into the study group and the reference group,with 49 cases in each group.The reference group was treated with albumin-bound paclitaxel,while the study group was treated with camrelizumab+albumin-bound paclitaxel.After treatment,the overall efficacy was evaluated,and the disease control rate was calculated.Serum tumor markers[high mobility group protein B1(HMGB1),squamous cell carcinoma associated antigen(SCCA)],PD-1 and PD-L1 levels were detected by enzyme-linked immunosorbent assay(ELISA)before and after treatment.Immunohistochemistry was used to detect the microvascular density(MVD)and the expression levels of PD-1 and PD-L1 in esophageal carcinoma tissue.The percentages of CD3+,CD4+and CD8+cells were detected by flow cytometry.Patients were followed up and adverse reactions and survival were recorded.Results The disease control rate was higher in the study group than that of the control group(P＜0.05).There were no significant differences in all indexes between the groups before treatment(P＞0.05).After treatment,HMGB1 level,SCCA level,MVD,serum PD-1 level and tumor tissue high expression rate of PD-1 were all decreased in the two groups,while serum PD-L1 level,PD-L1 high expression rate and percentages of CD3+,CD4+and CD8+cells were all increased(P＜0.05).Except there was no significant difference in MVD between groups,the other indexes were significantly changed in the study group compared with the control group(P＜0.05).There was no significant difference in the incidence of adverse reactions between the two groups(P＞0.05).The progression-free survival rate was higher in the study group than that of the control group(P＜0.05).Conclusion Carrellizumab combined with albumin-bound paclitaxel in the treatment of locally advanced esophageal cancer can help control lesion progression and inhibit angiogenesis,with good safety.

With the vigorous promotion of organ donation after citizen death in China, increased utilization of marginal livers, and continuous expansion of hepatocellular carcinoma indications for liver transplantation, innovations in techniques such as auxiliary liver transplantation, pediatric liver transplantation, laparoscopic liver transplantation, magnetic liver transplantation,and non-ischemic liver transplantation have significantly improved the number of liver transplantation surgery performed, patient survival rates, and graft survival rates in China, while complication rates have gradually decreased. As such,liver transplantation in China has reached leading or advanced levels internationally. In the new development context,Chinese liver transplantation faces new opportunities and challenges for development. Evolutions in basic diseases of transplant recipients and tumor classifications of will further broaden the population eligible for transplantation and introduce new demands for liver transplantation procedures. Emerging technologies including artificial organs, xenotransplantation,and artificial intelligence are bringing prospects for advancing liver transplantation. Looking ahead, the progression of liver transplantation will go beyond prioritizing patient survival rates and graft survival rates alone, instead emphasizing improved quality of life for transplant recipients post-surgery to an even greater extent.

With the vigorous promotion of organ donation after citizen death in China, increased utilization of marginal livers, and continuous expansion of hepatocellular carcinoma indications for liver transplantation, innovations in techniques such as auxiliary liver transplantation, pediatric liver transplantation, laparoscopic liver transplantation, magnetic liver transplantation,and non-ischemic liver transplantation have significantly improved the number of liver transplantation surgery performed, patient survival rates, and graft survival rates in China, while complication rates have gradually decreased. As such,liver transplantation in China has reached leading or advanced levels internationally. In the new development context,Chinese liver transplantation faces new opportunities and challenges for development. Evolutions in basic diseases of transplant recipients and tumor classifications of will further broaden the population eligible for transplantation and introduce new demands for liver transplantation procedures. Emerging technologies including artificial organs, xenotransplantation,and artificial intelligence are bringing prospects for advancing liver transplantation. Looking ahead, the progression of liver transplantation will go beyond prioritizing patient survival rates and graft survival rates alone, instead emphasizing improved quality of life for transplant recipients post-surgery to an even greater extent.

Objective:To analyze the types and distribution of pathogenic variants for neonatal genetic diseases in Huzhou, Zhejiang Province.Methods:One thousand neonates (48 ~ 42 h after birth) born to Huzhou region were selected as the study subjects. Dry blood spot samples were collected from the newborns, and targeted capture high-throughput sequencing was carried out for pathogenic genes underlying 542 inherited diseases. Candidate variants were verified by Sanger sequencing.Results:Among the 1 000 newborns, the male to female ratio was 1.02 : 1.00. No pathogenic variants were detected in 253 cases, whilst 747 cases were found to carry at least one pathogenic variant, which yielded a carrier rate of 74.7%. The most frequently involved pathogenic gene was FLG, followed by GJB2, UGT1A1, USH2A and DUOX2. The variants were classified as homozygous, compound heterozygous, and hemizygous variants. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), 213 neonates were verified to have carried pathogenic and/or likely pathogenic variants, with a positive rate of 21.3%. The most commonly involved genes had included UGT1A1, FLG, GJB2, MEFV and G6PD. Conclusion:Newborn screening based on high-throughput sequencing technology can expand the scope of screening and improve the positive predictive value. Genetic counseling based on the results can improve the patients′ medical care and reduce neonatal mortality and childhood morbidity, while provide assistance to family members′ health management and reproductive decisions.

Objective To predict and analyze the potential suitable areas of the Guangxi characteristic medicinal plant Zanthoxylum nitidum(Roxb.)DC.in China under climate change;To provide reference for ecological conservation and artificial cultivation of Zanthoxylum nitidum(Roxb.)DC.Methods Based on the 49 pieces of geographical distribution information of Zanthoxylum nitidum(Roxb.)DC.in China,19 environmental factors,and the maximum entropy(MaxEnt)model,a species distribution model was constructed to predict and analyze the potential growth areas of Zanthoxylum nitidum(Roxb.)DC.under current and future climate change.Results Under the current climate change,the potential suitable areas of Zanthoxylum nitidum(Roxb.)DC.in China mainly depended on 4 environmental factors:precipitation in the coldest season(Bio19),precipitation in the warmest season(Bio18),precipitation in the driest season(Bio17),and average annual temperature(Bio1).Under the future climate change,the center of mass of the high suitable areas of Zanthoxylum nitidum(Roxb.)DC.will mainly migrate to the northeast and northwest,and the migration distance will range from 13.63 km to 153.95 km.Compared with the present,the stable and unchanged high suitable areas of Zanthoxylum nitidum(Roxb.)DC.under future climate change will be reduced to about 21.68%,and mainly distributed in southwest Guangxi,and these stable and unchanged areas will become the first choice for semi-artificial planting of Zanthoxylum nitidum(Roxb.)DC.Conclusion The results are helpful to provide scientific basis for formulating reasonable ecological conservation and artificial cultivation strategies to reduce the impact of climate change on the population spread of Zanthoxylum nitidum(Roxb.)DC.