Recent clinical studies have shown that mutation of phosphatase and tensin homolog deleted on chromosome 10 (PTEN) gene in cancer cells may be associated with immunosuppressive tumor microenvironment (TME) and poor response to immune checkpoint blockade (ICB) therapy. Therefore, efficiently restoring PTEN gene expression in cancer cells is critical to improving the responding rate to ICB therapy. Here, we screened an adeno-associated virus (AAV) capsid for efficient PTEN gene delivery into B16F10 tumor cells. We demonstrated that intratumorally injected AAV6-PTEN successfully restored the tumor cell PTEN gene expression and effectively inhibited tumor progression by inducing tumor cell immunogenic cell death (ICD) and increasing immune cell infiltration. Moreover, we developed an anti-PD-1 loaded phospholipid-based phase separation gel (PPSG), which formed an in situ depot and sustainably release anti-PD-1 drugs within 42 days in vivo. In order to effectively inhibit the recurrence of melanoma, we further applied a triple therapy based on AAV6-PTEN, PPSG^@anti-PD-1 and CpG, and showed that this triple therapy strategy enhanced the synergistic antitumor immune effect and also induced robust immune memory, which completely rejected tumor recurrence. We anticipate that this triple therapy could be used as a new tumor combination therapy with stronger immune activation capacity and tumor inhibition efficacy.

Background and objective Transcription factor(TF)can bind specific sequences that either promotes or represses the transcription of target genes,and exerts important effects on tumorigenesis,migration,invasion.Staphylococcal nuclease-containing structural domain 1(SND1),which is a transcriptional co-activator,is considered as a promising target for tumor therapy.However,its role in lung adenocarcinoma(LUAD)remains unclear.This study aims to explore the role of SND1 in LUAD.Methods Data from The Cancer Genome Atlas(TCGA),Gene Expression Omnibus(GEO),Clinical Pro-teomic Tumor Analysis Consortium(CPTAC),and Human Protein Atlas(HPA)database was obtained to explore the associa-tion between SND1 and the prognosis,as well as the immune cell infiltration,and subcellular localization in LUAD tissues.Furthermore,the functional role of SND1 in LUAD was verified in vitro.EdU assay,CCK-8 assay,flow cytometry,scratch assay,Transwell assay and Western blot were performed.Results SND1 was found to be upregulated and high expression of SND1 is correlated with poor prognosis of LUAD patients.In addition,SND1 was predominantly present in the cytoplasm of LUAD cells.Enrichment analysis showed that SND1 was closely associated with the cell cycle,as well as DNA replication,and chro-mosome segregation.Immune infiltration analysis showed that SND1 was closely associated with various immune cell popula-tions,including T cells,B cells,cytotoxic cells and dendritic cells.In vitro studies demonstrated that silencing of SND1 inhib-ited cell proliferation,invasion and migration of LUAD cells.Besides,cell cycle was blocked at G,phase by down-regulating SND1.Conclusion SND1 might be an important prognostic biomarker of LUAD and may promote LUAD cells proliferation and migration.

Objective:To analyze the distribution of different SF3B1 genotypes in patients with myelodysplastic syndromes (MDS) and its prognostic value.Methods:Totally, 377MDS patients who were initially diagnosed in the First Affiliated Hospital of Nanjing Medical University from January 2014 to January 2022 were included in the retrospective analysis.The patients were divided into three different groups according to mutation stcote of SF3B1, including 317 patients with SF3B1 wild type (SF3B1 WT) (214 males and 103 females, 63(49, 71) years old),39 patients with SF3B1 K700E mutation(SF3B1 K700E(17 males and 22 females, 65(52, 73)years old)) and 21 patients with SF3B1 non-K700E mutation(SF3B1 non-K700E)(13 males and 8 females, 67(63, 73) years old). MDS-related 20 gene mutations were detected using targeted sequencing technology; Survival curves were constructed by the Kaplan-Meier method; Cox proportional hazards model was established to evaluate different factors at diagnosis on survival by univariate and multivariate analyses.. Results:Compared with SF3B1 non-K700E patients, SF3B1 K700E patients had a higher median absolute neutrophil count ( P=0.002) and were likely to be in the low/int-1 International Prognostic Scoring System (IPSS) categories ( P=0.023). A 20-gene targeted sequencing analysis showed that, compared with SF3B1 WT patients, SF3B1 K700E patients were associated with lower frequency of ASXL1 and U2AF1 mutations ( P=0.018 and P=0.003); while compared with SF3B1 non-K700E patients, the frequency of ASXL1 mutation was significantly lower in SF3B1 K700E cases ( P=0.029). Patients with SF3B1 K700E had better overall survival (OS) in comparison with SF3B1 WT and SF3B1 non-K700E in MDS patients ( P<0.001 and P=0.045, respectively). In comparison with SF3B1 WT patients, SF3B1 MUT patients had more favorable OS and progression-free survival (PFS) in MDS without excess blasts ( P<0.001 and P<0.001, respectively), but no significant difference was found in MDS with excess blasts ( P>0.05). Compared with SF3B1 WT patients, SF3B1 K700E patients had superior OS and PFS in the int-1 IPSS category ( P=0.010 and P=0.013, respectively). By multivariable analysis, the presence of SF3B1 K700Ewas an independent predictor of superior OS ( HR=0.461,95% CI 0.262-0.811, P=0.007). Conclusion:SF3B1 K700E and SF3B1 non-K700E patients had significantly improved OS in comparison with SF3B1 WT MDS patients. Furthermore, SF3B1 K700E patients were associated with a better OS compared with SF3B1 non-K700E MDS patients. SF3B1 mutation could not overcome the poor prognostic effect of excess blasts, which highlights the importance of the SF3B1 mutation subtype in risk assessment of MDS without excess blasts.

Due to the insufficient long-term protection and significant efficacy reduction to new variants of current COVID-19 vaccines, the epidemic prevention and control are still challenging. Here, we employ a capsid and antigen structure engineering (CASE) strategy to manufacture an adeno-associated viral serotype 6-based vaccine (S663V-RBD), which expresses trimeric receptor binding domain (RBD) of spike protein fused with a biological adjuvant RS09. Impressively, the engineered S663V-RBD could rapidly induce a satisfactory RBD-specific IgG titer within 2 weeks and maintain the titer for more than 4 months. Compared to the licensed BBIBP-CorV (Sinopharm, China), a single-dose S663V-RBD induced more endurable and robust immune responses in mice and elicited superior neutralizing antibodies against three typical SARS-CoV-2 pseudoviruses including wild type, C.37 (Lambda) and B.1.617.2 (Delta). More interestingly, the intramuscular injection of S663V-RBD could overcome pre-existing immunity against the capsid. Given its effectiveness, the CASE-based S663V-RBD may provide a new solution for the current and next pandemic.

Objective : To investigate the expression , clinical significance , progression , and prognosis of miR⁃381 ⁃3p in acute myeloid leukemia (AML) , as well as its impact on AML cell proliferation and apoptosis , in order to provide theoretical basis for the treatment of AML. Methods : Bioinformatics analysis was used to identify differentially expressed miRNAs , clinical data and blood samples of AML patients were collected , and the expression levels of miRNAs in the bone marrow fluid of the included patients were measured to further elucidate the relationship between miRNAs and AML. The included patients were followed up to calculate overall survival (OS) and disease⁃free survival (DFS) ; AML cells were cultured in vitro , miR⁃381 ⁃3p plasmids were constructed , miR⁃381 ⁃3p was overexpressed and miR⁃381 ⁃3p was knocked down in AML , and they were divided into five groups : control , miR⁃381 mimics , mimics NC , miR⁃381 inhibitor , inhibitor NC . The proliferation and apoptosis of AML cells were detected using CCK⁃8 and flow cytometry . Results : Differentially expressed miRNAs were identified using bioinforDifferentially expressed miRNAs were identified using bioinfor included . The expression level of miR⁃381 in AML patients was lower than that in the control group , and all FAB subtypes were lower than those in the normal group . The expression level of miR⁃381 was not related to the age , gender , peripheral blood leukocytes , lymphocytes , and FAB typing of AML patients , and the OS and DFS of miR⁃381 patients with high expression were significantly prolonged , with statistically significant differences . In vitro experiments had shown that knocking down miR⁃381 could inhibit apoptosis and promote proliferation of AML cells . Overexpression of miR⁃381 could promote apoptosis and inhibit proliferation of AML cells . Conclusion MiR⁃381 ⁃ 3p is low expressed in AML patients , and its overexpression can significantly prolong OS and DFS . miR⁃381 ⁃3p can promote apoptosis of AML cells , inhibit proliferation , and may become a targeted molecule for the treatment of AML.

Objective:To assess the efficacy of induction chemotherapy followed by allogeneic hematopoietic stem cell transplantation (allo-HSCT) in the treatment of FLT3-ITD + acute myeloid leukemia (AML) with normal karyotype. Methods:The clinical data of FLT3-ITD + AML patients with normal karyotype in the First Affiliated Hospital of Nanjing Medical University from Jan 2018 to March 2021 were retrospectively analyzed. Results:The study included 49 patients with FLT3-ITD +AML, 31 males, and 18 females, with a median age of 46 (16-59) years old. All patients received induction chemotherapy, and 24 patients received sequential allo-HSCT (transplantation group) . The median follow-up time was 465 days, the one-year overall survival (OS) from diagnosis was (70.0 ± 7.4) %, and one-year disease-free survival (DFS) was (70.3±7.4) %. The one-year OS was significantly different between the transplantation group and the non-transplantation group [ (85.2 ± 7.9) % vs (52.6 ± 12.3) %, P=0.049]. but one-year DFS [ (84.7 ± 8.1) % vs (55.2 ± 11.9) %, P=0.061] was not. No significance was found in one-year OS between patients with low-frequency and high-frequency FLT3-ITD + ( P>0.05) . There were 12 patients with high-frequency FLT3-ITD + in the transplantation and the non-transplantation groups, respectively. The one-year OS [ (68.8 ± 15.7) % in the transplantation group vs (26.2 ± 15.3) % in the non-transplantation group, P=0.027] and one-year DFS [ (45.5 ± 21.3) % in the transplantation group vs (27.8±15.8) % in the non-transplantation group, P=0.032] were significantly different between the two groups. Conclusion:Induction chemotherapy followed by allo-HSCT can enhance the prognosis of FLT3-ITD + patients, particularly those with FLT3-ITD high-frequency mutation.

Objective:To investigate the related factors of nonalcoholic steatohepatitis in children with obesity.Methods:A retrospective analysis was performed on 91 children with obesity who visited the Pediatric Obesity Clinic in the Affiliated Hospital of Hangzhou Normal University from July 2020 to January 2021. The 91 children with obesity were divided into two groups: with or without nonalcoholic steatohepatitis. Age, gender, body mass index, blood 25 hydroxyvitamin D3, fasting blood glucose, total cholesterol, triglyceride, low density lipoprotein, fasting insulin, control attenuation parameter and liver hardness value of in the 2 groups were recorded. Univariate analysis of the clinical data of the two groups was performed, and the clinical data with statistically significant differences between the two groups were included in binary logistic regression analysis to explore the related factors of nonalcoholic steatohepatitis in children with obesity. And then the receiver operating characteristic curve (ROC curve) was drawn on the relevant factors.Results:The children with obesity received treatment at the age of 6 years 1 month to 14 years 11 months, the male to female ratio was 1.33∶1. And 13 children (14.3%) were diagnosed with non-alcoholic steatohepatitis, of 46 nonalcoholic fatty liver disease, 9 were male and 4 were female. Univariate analysis showed that there were significant differences in gender, age, fasting insulin, control attenuation parameter and liver hardness value between the two groups (all P<0.05). Binary logistic regression analysis showed that control attenuation parameter ( OR=1.022, 95% CI: 1.001-1.041) and liver hardness value ( OR=1.689, 95% CI: 1.077-2.648) were the related factors of nonalcoholic steatohepatitis in children with obesity (both P<0.05). The area under the curve (AUC) values of control attenuation parameter and liver hardness value for predicting nonalcoholic steatohepatitis in children with obesity was 0.840 (95% CI: 0.748-0.931) and 0.794 (95% CI: 0.672-0.915), respectively. Conclusion:Control attenuation parameter and liver hardness value are correlated with nonalcoholic steatohepatitis in children with obesity with certain diagnostic value.

Objective:To investigate the appropriate age for booster doses of hepatitis B vaccine in children aged 0-14.Methods:Retrospective study.A total of 3 118 children aged 0-14 years who underwent quantitative serological marker testing for hepatitis B virus at the Affiliated Hospital of Hangzhou Normal University from January 2015 to October 2021 were recruited in this analysis.There were 1 702 males and 1 416 females, with a male to female ratio of 1.20∶1.00.Children were divided into 15 groups according to their age, and the classifying interval was 1 year.The hepatitis B virus surface antibody (Anti-HBs) titer was quantified by chemiluminescent microparticle immunoassay.The Anti-HBs positivity rates and hepatitis B immune response among groups of different sexes and age were compared by the chi- square test and rank- sum test, respectively. Results:A total of 3 118 children were investigated.The titer and effective response rate of Anti-HBs decreased gradually with age.The difference in the titer and effective response rate of Anti-HBs was statistically significant among groups of different age (all P<0.01), but not significant between males and females (all P>0.05). The median titer of Anti-HBs in children aged above 3 years was 58.49 IU/L(0-1 001.00 IU/L). About 59.1% (1 477/2 497 cases) of children aged 3 years and above had no immune response or low immune response (i.e., the titer of Anti-HBs was below 100 IU/L). Conclusions:The immune protective effect of the hepatitis B vaccine decreases year by year in children who have received the standardized vaccine, and the vaccine has poor protective effect on most children aged 3 years and above.Therefore, booster dose vaccination for preventing hepatitis B is necessary for children aged 3 and above.

Objective: To study the miR-1285 through Yes-associated protein 1(YAP1) on the proliferation, apoptosis and mechanism of chronic myelogenous leukemia K562 cells. Methods: The plasmid of miR-1285 was constructed to make K562 cells overexpress miR-1285 and knock down miR-1285. They were divided into four groups: miR-1285 mimics, mimics control, miR-1285 inhibitor, and inhibitor control; qRT-PCR was used to quantitatively analyze the expression of miR-1285 of these four groups. The expression level of miR-1285 in the group was used to verify the transfection efficiency. The apoptosis and proliferation of the four groups were detected by CCK-8 and Annexin V-FITC; the changes of YAP and its downstream molecule epidermal growth factor receptor(EGFR) were detected by Western blot, and apoptosis related molecules BAX, Bcl-2 protein expression was detected. Results: After knock down miR-1285, it can promote the proliferation of K562 cells and inhibit apoptosis. The expression of YAP increased, the expression of downstream molecule EGFR increased correspondingly, the expression of apoptosis-related molecule BAX decreased, and the expression of Bcl-2 increased; After expressing miR-1285, K562 cell proliferation decreased, apoptosis increased, YAP expression decreased, downstream molecule P-EGFR expression decreased, apoptosis molecule BAX expression increased, and Bcl-2 expression decreased. Conclusion miR-1285 can inhibit the proliferation and promote apoptosis of K562 cells of chronic myelogenous leukemia by inhibiting the expression of YAP. It is expected to become a targeted molecule for the treatment of chronic myelogenous leukemia.

Objective To evaluate the efficacy,bleeding profile and safety of low-dose levonorgestrel-releasing intrauterine system (LNG-IUS 8) in Chinese healthy women of childbearing age.Methods A multi-center,open-label,single-arm clinical trial conducted at 16 centres in China enrolled 773 healthy women of childbearing age (mean age 31.6 years old,range 18 to 40 years old),who demanded contraception,from April 2006 to June 2013.All women placed LNG-IUS 8 for 3 years and then been followed up at 3,6,9,12,18,24,30,36 months.The efficacy variables including pregnancy rate and expulsion rate were analyzed using life table,while observing adverse events (AE) to evaluate the safety.The bleeding profile happened during the study was assessed using 90-day reference intervals (World Health Organization criteria).Results Eight pregnancies occurred among 773 women,resulting in a overall Pearl index of 0.42 per 100 women years.The 3-year cumulative pregnancy rate was 0.37 per 100 women years and the 3-year cumulative expulsion rate was 1.99 per 100 women years.The number of women with bleeding/spotting reduced and the bleeding/spotting days declined over time.Totally 219 AE were reported related to LNG-IUS 8 placements.The most common AE were vaginal bleeding (8.2％,63/773)and the ovarian cyst (6.2％,52/773).LNG-IUS 8 had an improving effect on dysmenorrhea that the percentage of women with dysmenorrhea as well as the days of dysmenorrhea decreased over time.The percentage of women satisfied or very satisfied with LNG-IUS 8 was 87.2％ (622/713).Conclusion LNG-IUS 8 is highly effective and safe for Chinese healthy women of childbearing age.