Objective To investigate the antigen presentation characteristics of dendritic cells(DC)and B cells in cardiac grafts.Methods The heart of BALB/c mice was transplanted into the abdominal cavity of C57BL/6J mice.CD45+cells in the heart graft were extracted and sorted by flow cytometry at postoperative 5 d,and single cell RNA sequencing was performed.Taking DC and B cell subsets in cardiac grafts as the main study cells,the changing trend,antigen presenting ability and intercellular communication with T cells after heart transplantation were analyzed by bioinformatics analysis and flow cytometry.Gene ontology(GO)function enrichment difference analysis was adopted to prove the specific function and the reliability annotation of cell subsets.Results Germinal center-like B cell(GC-L B)was the B cell subset with the largest increase in quantity during the acute rejection phase,accounting for 87％.Classical DC(cDC)2 was the only DC subset with a significant increase in quantity during acute rejection of heart transplantation,accounting for 44％of DC subset,and it occupied the highest communication intensity with T cells after heart transplantation.Mononucleated DC(moDC)and memory B cell(MBC)were the main transmitters of T cell input signals in non-transplanted hearts,whereas transformed into cDC2 and GC-L B during the acute rejection phase.Among them,MBC and GC-L B were the main sources of T cell input signals in non-transplanted hearts and heart grafts.Conclusions Compared with DC,B cells occupy a higher number and weight in the intercellular communication with T cells in non-transplanted hearts and heart grafts,prompting that the antigen presenting activity of B cells is more active and stronger than DC in the early stage of acute rejection of heart transplantation.

Objective By population survey,to explore the epidemiological characteristics of gastric precancerous lesions in Lishui District of Nanjing and provide objective basis for the prevention and treatment of early gastric cancer.Methods From July 2021 to December 2022,21 977 patients who received endoscopy and/or 13C-UBT in Lishui District People's Hospital and 6 medical community units in Nanjing City were retrospectively analyzed for demography characteristics,detection rate of gastric precancerous lesions,and H.Pylori infection rate.Results(1)590 cases of gastric precancerous lesions were detected(detection rate 2.68%);(2)The total detection rate of precancerous lesions and three pathological types in males were all higher than those in females(all P<0.001);(3)The minimum age for the total detection rate of precancerous lesions in males and the mini-mum age for each pathological type were lower than in females(P<0.001,0.009,0.005,0.002);(4)The popu-lation total H.pylori infection rate was 23.10%,the H.pylori infection rate in patients with precancerous lesions was higher than that in non-precancerous lesions(P<0.001),both H.pylori infection rate of male and female in precancerous lesions were all higher than those of non-precancerous lesions of the same sex(all P<0.001),in addition,the H.pylori infection rate of male whether in precancerous or non-precancerous lesions was higher than that of female(all P<0.001);(5)The precancerous lesions detection rate in male,female,and the overall age range of 20～29 to 70～79 years is positively correlated with age growth(P<0.001),and rapidly decreases after the age of 79,the of H.pylori infection rate was also positively correlated with age growth(P<0.001),and the trend of age change(P<0.001)was parallel to the precancerous lesions detection rate.Conclusions The detec-tion rate of gastric precancerous lesions in this region is above the average level in China;the total H.pylori infec-tion rate is at a relatively low level in China;the H.pylori infection rate is parallel to the age trend of the detection rate of gastric precancerous lesions,and increases with age.

Objective To isolate endophytic fungi from Angelica sinensis and evaluate the bioactivity of their secondary metabolites.Methods Angelica sinensis and rhizosphere soil were utilized as materials.The tissue homogenization method was employed with six diverse culture media to isolate endophytic fungi.The antibacterial activity of secondary metabolites was gauged using a 96-well plate assay,while UV spectrophotometry was used to evaluate the inhibitory activity of four enzymes.Results A total of 153 fungal strains were isolated and purified from Angelica sinensis roots,stems,leaves,and soil.The samples exhibited specific inhibitory activities against adenosine deaminase(ADA),β-lactamase,xanthine oxidase(XO),and tyrosinase(TYR),with rates of 45.83%,52.78%,51.39%and 55.56%,respectively.Furthermore,1.39%of the samples displayed wide-ranging inhibitory effects against four indicator bacteria.Strain 6B also showcased the lowest inhibitory concentration values of 62.5 and 7.81 μg/mL against Escherichia coli ATCC25922 and ATCC35218,respectively,signifying its potential research significance.Conclusion Angelica sinensis has abundant endophytic fungal resources and is a good source for discovering active compounds,demonstrating certain research value.

Objective To evaluate the role of preoperative serological indexes in predicting long-term survival and tumor recurrence of hepatocellular carcinoma (HCC) patients after liver transplantation, aiming to explore its significance in expanding the Milan criteria. Methods Clinical data of 669 recipients undergoing liver transplantation for HCC were retrospectively analyzed. The optimal cut-off value was calculated by the receiver operating characteristic (ROC) curve. The risk factors affecting the overall survival and recurrence-free survival rates of HCC patients after liver transplantation were identified by univariate and multivariate regression analyses. The correlation between preoperative serum liver enzymes and pathological characteristics in HCC patients was analyzed. The predictive values of alpha-fetoprotein (AFP) combined with γ -glutamyl transferase (GGT) and different liver transplant criteria for the survival and recurrence of HCC patients after liver transplantation were compared. Results Exceeded Milan criteria, total tumor diameter (TTD) > 8 cm, AFP > 200 ng/mL and GGT > 84 U/L were the independent risk factors for the overall survival and recurrence-free survival rates of HCC patients after liver transplantation (all P < 0.05). Correlation analysis showed that preoperative serum GGT level was correlated with TTD, number of tumor, venous invasion, microsatellite lesions, capsular invasion, tumor, node, metastasis (TNM) stage, Child-Pugh score and exceeded Milan criteria (all P < 0.05). Milan-AFP-GGT-TTD (M-AGT) criteria were proposed by combining Milan criteria, TTD with serum liver enzyme indexes (AFP and GGT). The 5-year overall survival and recurrence-free survival rates of HCC recipients who met the M-AGT criteria (111 cases of exceeded Milan criteria) were significantly higher than those who met Hangzhou criteria (both P < 0.05), whereas had no significant difference from their counterparts who met the University of California at San Francisco (UCSF) criteria (both P > 0.05). Conclusions Preoperative serological indexes of AFP and GGT could effectively predict the long-term survival and tumor recurrence of HCC patients after liver transplantation. Establishing the M-AGT criteria based on serological indexes contributes to expanding the Milan criteria, which is convenient and feasible.

Objective:To validate the feasibility of the gamma analysis method in the study of prescription dose conversion between logistic nanodosimetry model (LNDM) and microdosimetric kinetic model (MKM) basing on the Chinese self-developed model LNDM by applying clinical experiences of National Institute of Radiological Science (NIRS).Methods:Physical dose distributions derived from the MKM- and LNDM-based carbon ion treatment plans were compared via the method of gamma analysis under the open-source treatment planning platform matRad. In this way, the prescribed dose conversion factor between the MKM- and LNDM-based treatment plans was obtained. Using water phantoms, the influence of geometric shape, size, depth of target volume (TV), prescribed dose and field setting on the conversion factor was investigated comprehensively. Moreover, preliminary verification of the acquired conversion factor was conducted on the C-shape model and a case of liver cancer patient.Results:The conversion factor depended on the field setting rather than the TV shape. Under the condition of single field, the conversion factor was positively correlated with the size and depth of TV, and the prescribed dose. Moreover, the conversion factor was successfully verified using the C-shape model and the patient with liver cancer, where the gamma passing rates (2%/2 mm) of the physical dose distribution generated by the MKM and LNDM treatment plans were 92.79% and 91.19%, respectively.Conclusions:The conversion factors (f=D LNDM/D MKM) obtained in this study might provide guidance for the prescribed dose setting during the carbon ion treatment planning based on the LNDM. Besides, the gamma analysis method could be used for the study of the prescribed dose conversion between different models.

The identification of tumor-related microRNAs(miRNAs)exhibits excellent promise for the early diag-nosis of cancer and other bioanalytical applications.Therefore,we developed a sensitive and efficient biosensor using polyadenine(polyA)-mediated fluorescent spherical nucleic acid(FSNA)for miRNA analysis based on strand displacement reactions on gold nanoparticle(AuNP)surfaces and electrokinetic signal amplification(ESA)on a microfluidic chip.In this FSNA,polyA-DNA biosensor was anchored on AuNP surfaces via intrinsic affinity between adenine and Au.The upright conformational polyA-DNA recognition block hybridized with 6-carboxyfluorescein-labeled reporter-DNA,resulting in fluores-cence quenching of FSNA probes induced by AuNP-based resonance energy transfer.Reporter DNA was replaced in the presence of target miRNA,leading to the recovery of reporter-DNA fluorescence.Sub-sequently,reporter-DNAs were accumulated and detected in the front of with Nafion membrane in the microchannel by ESA.Our method showed high selectivity and sensitivity with a limit of detection of 1.3 pM.This method could also be used to detect miRNA-21 in human serum and urine samples,with re-coveries of 104.0％-113.3％and 104.9％-108.0％,respectively.Furthermore,we constructed a chip with three parallel channels for the simultaneous detection of multiple tumor-related miRNAs(miRNA-21,miRNA-141,and miRNA-375),which increased the detection efficiency.Our universal method can be applied to other DNA/RNA analyses by altering recognition sequences.

Internet hospitals played a huge role in meeting patients’ medical needs during the COVID-19 pandemic. As a new service model, it also brings new challenges to medical ethics. This paper analyzed the three key elements of behavior: actors, behavior process and action result, and found that apart from ethical challenges such as medical security, data security and privacy protection, different types of Internet hospitals show diversity due to different actors involved. The first type of Internet hospital represented the conflict of interest of doctors brought by the integration of offline and online working modes. The second type showed the challenge of fairness in medical treatment brought by the demand of personalized service provided by medical staff in public medical institutions. The third type manifested the medical security challenges brought by doctors with varying technical levels in an open platform environment. Combined with the current ethical governance of science and technology and the digital transformation of society, this paper put forward some countermeasures and suggestion, such as further clarifying the boundaries of rights and responsibilities of relevant subjects of Internet platform, formulating the code of conduct requirements for medical staff in Internet hospitals, and supervising doctors’ medical practice in Internet hospitals.

Objective: To study the overexpression of protein phosphatase 2 regulatory subunit B''α gene effects on the proliferation and invasion of hepatoma cells. Methods: Immunohistochemistry method was used to analyze the expression of PPP2R3A in cancerous and paracancerous tissues. Hepatocellular carcinoma cell lines (Huh-7 and HepG2) with stably overexpressing PPP2R3A were constructed by lentiviral vector. Biological behavioral transition in hepatocellular carcinoma cell proliferation, cell cycle, apoptosis, invasion and metastasis were detected by cell counting kit-8 assay (CCK-8), flow cytometry, and transwell assay. A subcutaneous nude tumor mice model was constructed to validate the growth of hepatoma cells. Two independent sample t-tests were used to compare the groups. Results: The expression of PPP2R3A gene in human hepatocarcinoma tissues was higher than paracancerous tissues. The absorbance (A value) of hepatoma cells was increased (P < 0.05) after overexpression of PPP2R3A gene. The transition from G1-to-S phase was significantly increased i.e., the G1 phase of the cell cycle was reduced (Huh-7: t = 3.04, P = 0.0384; HepG2: t = 4.06, P = 0.0153), while the S phase was increased (Huh-7: t = 3.47, P = 0.0255; HepG2: t = 4.46, P = 0.0112). Early apoptotic rate was decreased (Huh-7: t = 7.34, P = 0.0018; HepG2: t = 4.06, P = 0.0153). The number of Huh-7 cells migrating to the lower chamber was increased (t = 3.18, P = 0.0334), and after the use of matrigel the number of cells reaching to the lower chamber was also increased (t = 2.84, P = 0.0464). The results of animal experiments showed that the subcutaneous tumor growth (t = 4.31, P = 0.0035) was significantly overexpressed in nude mice group. The results of Western blot showed that the expression of PARP and P53 protein in the spliced ​​forms decreased, while the accumulation of β-catenin protein in the liver cancer cells was increased. Conclusion Overexpressed PPP2R3A gene may promote proliferation, migration and invasion ability, inhibit apoptosis, induce G1/S phase transition, and participate in the biological behavior of hepatoma cells.

Objective To explore the prognostic role of laminin (LN ) as a tumor biological marker in predicting the recurrence of hepatocellular carcinoma (HCC) related with HBV infection after liver transplantation (LT ) .Methods Tissue samples from 251 HBV-related HCC patients undergoing LT were immunohistochemically stained with anti-LN antibody .The relevant prognostic factors were analyzed using Spearman's rank test , Kaplan-Meier method , log-rank test and multivariate step-wise Cox regression analysis .Results The expressions of LN in tumor tissues were significantly positively correlated with tumor number (P=0 .00) ,microsatellite (P=0 .02) ,venous invasion (P=0 .048) ,pTNM tumor stage (P=0 .00) ,pre-LT serum α-fetoprotein (AFP) level (P=0 .00) ,HBV DNA level (P= 0 .02) ,HBeAg level (P= 0 .02) and tumor recurrence (P= 0 .00) respectively .Significant differences existed in 1/3/5-year overall survival or tumor recurrence-free survival rate post-LT among LN different expression (-,+ ,≥ + + ) in HBV-related HCC patients (P< 0 .05 ) . Multivariate analysis indicated that LN was a significantly independent predictor in predicting poor tumor recurrence-free survival post-LT (P=0 .01) .Conclusions LN may be a feasible marker in predicting HCC recurrence post-LT for HBV-related HCC patients .

Gene therapy represents a promising treatment for the Alzheimer׳s disease (AD). However, gene delivery specific to brain lesions through systemic administration remains big challenge. In our previous work, we have developed an siRNA nanocomplex able to be specifically delivered to the amyloid plaques through surface modification with both CGN peptide for the blood-brain barrier (BBB) penetration and QSH peptide for -amyloid binding. But, whether the as-designed nanocomplex could indeed improve the gene accumulation in the impaired neuron cells and ameliorate AD-associated symptoms remains further study. Herein, we prepared the nanocomplexes with an siRNA against -site amyloid precursor protein-cleaving enzyme 1 (BACE1), the rate-limiting enzyme of A production, as the therapeutic siRNA of AD. The nanocomplexes exhibited high distribution in the A deposits-enriched hippocampus, especially in the neurons near the amyloid plaques after intravenous administration. In APP/PS1 transgenic mice, the nanocomplexes down-regulated BACE1 in both mRNA and protein levels, as well as A and amyloid plaques to the level of wild-type mice. Moreover, the nanocomplexes significantly increased the level of synaptophysin and rescued memory loss of the AD transgenic mice without hematological or histological toxicity. Taken together, this work presented direct evidences that the design of precise gene delivery to the AD lesions markedly improves the therapeutic outcome.