Bacteria have posed a threat to human health,and the emergence of super bacteria has made it more difficult to cure bacterial infections in clinical practice.Currently,vaccines are one of the effective means of preventing bacterial infections.With the rapid development of cutting-edge technologies in recent years in such disciplines as biology,medicine,and materials science,various innovative strategies have been provided for vaccine research and preparation.This article summarizes the status quo and prospects of innovative vaccines for treating bacterial infections in recent years,including subunit vaccines,mRNA vaccines and attenuated live vaccine in the hopes of providing data for subsequent development and research of bacterial vaccines.

Background::Rapid initiation of antiretroviral therapy (ART) is recommended by guidelines, however, real-world studies of same-day initiation of ART in China are limited, and an optimal treatment regimen has yet to be identified. The study aims to provide a realistic reference for rapid initiation of ART.Methods::We retrospectively analyzed the clinical data of treatment-na?ve people with human immunodeficiency virus (PWHs) who were diagnosed and prescribed same-day ART initiation from January 1, 2021 to December 31, 2022 at Chongqing Public Health Medical Center. PWHs voluntarily chose an ART regimen that divided them into two groups: National Free Antiretroviral Treatment Program (NFATP)-recommended regimens group (2 nucleoside reverse transcriptase inhibitors + non-nucleoside reverse transcriptase inhibitors/protease inhibitors) and bictegravir/emtricitabine/tenofovir alafenamide (BIC/FTC/TAF) group. The primary endpoint was the virological outcome of the two groups for same-day ART initiation at 24 weeks and 48 weeks. The secondary endpoints included changes in CD4 counts, maintenance of the original ART regimen at 48 weeks, and lipid levels and renal function at 48 weeks.Results::A total of 255 PWHs were included in the study, including 131 (51.4%) in the NFATP group and 124 (48.6%) in the BIC/FTC/TAF group. The overall virological suppression rates at 24 weeks and 48 weeks were 78.2% (165/211) and 95.4% (207/217), respectively. At 24 weeks, the virologic suppression rate in the NFATP group was lower than that in the BIC/FTC/TAF group (65.3% [66/101] vs. 90.0% [99/110], P <0.001). The median increase in the CD4 count was 198.0 (126.0-300.0) cells/μL at 24 weeks, with 182.0 (108.0-245.0) cells/μL in the NFATP group and 219 (132.0-316.0) cells/μL in the BIC/FTC/TAF group ( P = 0.035). At 48 weeks, there was no significant difference in the virological suppression rate or CD4 count between the groups. The 48-week initial ART regimen retention rates and treatment retention rates were significantly higher in the BIC/FTC/TAF group than in the NFATP group (91.1% (113/124) vs. 71.8% (94/131), 99.2% (118/119) vs. 93.0% (120/129), respectively). In terms of safety, there were no significant changes from baseline in levels of creatinine, estimated glomerular filtration rate (eGFR), or lipids in either group at 48 weeks. Conclusions::ART initiation on the day of diagnosis is effective, safe, and feasible, with satisfactory rates of virologic suppression, 48-week initial ART regimen retention rates, and treatment retention rates in treatment-na?ve PWHs. In our study, the early virologic suppression rate, CD4 cell counts, and treatment retention of the BIC/FTC/TAF regimens were significantly better than those of the NFATP regimens.

Objective To determine the neurotoxic effects of amyloid beta 42(Aβ42)oligomers and investigate the mechanism of their induction of Alzheimer's disease(AD)-like pathogenesis in neuronal cells.Methods Western blotting and transmission electron microscopy were used to identify the synthesized Aβ42 oligomers.In order to assess the impact of the oligomers,MTT assay was employed to measure cell viability in neuroblastoma cell line SH-SY5Y treated with 10μmol/L Aβ42 oligomers for 12 or 24 h,glutamatergic neurons derived from human embryonic stem cells(hESC)exposed to Aβ42 oligomers for 24,48,or 96 h,and corresponding control cells.TUNEL assay was utilized to assess the apoptosis of glutamatergic neurons.Additionally,immu-nofluorescence assay was applied to detect the changes in Aβ plaques and p-tau pathology.Results Western blotting displayed monomers and small-molecule aggregation(＜30 000)in our synthe-sized Aβ42 oligomers,and transmission electron microscopy showed that the synthesized oligomers were mainly in a shape of spherical particles.Treatment of 10 μmol/L Aβ42 oligomers for 12 and 24 h in SH-SY5Y cells significantly decreased cell viability when compared with the control cells[(70.89±2.54)％vs(100.00±2.02)％,(52.63±3.37)％vs(100.00±2.80)％,P＜0.05].The 10μmol/L oligomers treatment for 24,48 and 96 h also decreased cell viability in glutamatergic neu-rons(P＜0.05).The apoptotic rates was significantly higher in glutamatergic neurons after treat-ment for 48 and 96 h when compared to the control cells[(1.44±0.31)％vs(1.00±0.38)％,(1.75±0.64)％vs(1.00±0.31)％,P＜0.05].Furthermore,circular granular Aβ-positive plaque signals were observed in the glutamatergic neurons after treated with the oligomers for 24,48,and 96 h,but no such plaque signals were seen in the control cells.Additionally,glutamatergic neurons incu-bation with 10 μmol/L oligomers for 24 h resulted in tau hyperphosphorylation at the Thr231 site,with the average fluorescence intensity significantly higher than that in control cells(P＜0.05).Conclusion Aβ42 oligomers show toxic effects to both SH-SY5Y cells and glutamatergic neurons,and they can also induce glutamatergic neurons to produce AD pathology.

OBJECTIVE: To investigate the protective effects of total saponins from Panax japonicus (TSPJ) against high-fat dietinduced testicular Sertoli cell junction damage in mice. METHODS: Forty male C57BL/6J mice were randomized into normal diet group, high-fat diet group, and low-dose (25 mg/kg) and high-dose (75 mg/kg) TSPJ treatment groups (n=10). The mice in the normal diet group were fed a normal diet, while the mice in the other groups were fed a high-fat diet. After TSPJ treatment via intragastric administration for 5 months, the testes and epididymis of the mice were collected for measurement of weight, testicular and epididymal indices and sperm parameters. HE staining was used for histological evaluation of the testicular tissues and measurement of seminiferous tubule diameter and seminiferous epithelium height. The expression levels of ZO-1, occludin, claudin11, N-cadherin, E-cadherin and β-catenin in Sertoli cells were detected with Western blot, and the localization and expression levels of ZO-1 and β-catenin in the testicular tissues were detected with immunofluorescence assay. The protein expressions of LC3B, p-AKT and p-mTOR in testicular Sertoli cells were detected using double immunofluorescence assay. RESULTS: Treatment with TSPJ significantly improved high-fat diet-induced testicular dysfunction by reducing body weight (P < 0.001), increasing testicular and epididymal indices (P < 0.05), and improving sperm concentration and sperm viability (P < 0.05). TSPJ ameliorated testicular pathologies and increased seminiferous epithelium height of the mice with high-fat diet feeding (P < 0.05) without affecting the seminiferous tubule diameter. TSPJ significantly increased the expression levels of ZO-1, occludin, N-cadherin, E-cadherin and β-catenin (P < 0.05) but did not affect claudin11 expression in the testicular tissues. Immunofluorescence assay showed that TSPJ significantly increased ZO-1 and β-catenin expression in the testicular tissues (P < 0.001), downregulated LC3B expression and upregulated p-AKT and p-mTOR expressions in testicular Sertoli cells. CONCLUSION TSPJ alleviates high-fat diet-induced damages of testicular Sertoli cell junctions and spermatogenesis possibly by activating the AKT/mTOR signaling pathway and inhibiting autophagy of testicular Sertoli cells.
Male ; Animals ; Mice ; Mice, Inbred C57BL ; Testis ; Sertoli Cells ; beta Catenin ; Diet, High-Fat ; Occludin ; Proto-Oncogene Proteins c-akt ; Seeds ; Cadherins ; Intercellular Junctions

Immunotherapy has become a common means of cancer treatment. In immunotherapy, PD-1/PD-L1 inhibitors have significant efficacy. Cancer and various opportunistic infections are common complications in patients with AIDS. Owing to the special immune situation of these patients, AIDS is regarded as an exclusion standard in most clinical trials for cancer immunotherapy, conferring immunotherapy difficulty in treating patients with AIDS. The popularity of effective antiretroviral drugs has prolonged the lifetime of people with AIDS. Therefore, exploiting the opportunity of using immunotherapy in AIDS with cancer is urgent.

This article reviews the concept, importance, and assessment tools of nurse leadership. This article introduces the characteristics of each assessment tool and its application in different cultural backgrounds, so as to provide a reference for nurses to choose appropriate assessment tools, and to provide a basis for the development of assessment tools for nurse leadership in China.

Objective:To compare the activity difference of the high affinity humanized CD19 chimeric antigen receptor (CAR)-T cells and murine CD19 CAR-T cells.Methods:Peripheral venous blood T cells from 8 healthy volunteers were collected and infected with humanized and murine CD19 CAR lentivirus. Human and murine CD19 CAR-T cells were prepared and cell proliferation was detected by cell counting kit-8 (CCK-8) method. The cytotoxicity of CD3 + T cells, humanized and murine CD19 CAR-T cells to NALM-6 cells was detected by lactate dehydrogenase assay. Thirty BAL B/c nude mice transplanted with NALM-6 cells were randomly divided into 3 groups with 10 mice in each group and injected humanized CD19 CAR-T cells, mouse CD19 CAR-T cells and control CD3 + T cell via tail vein, respectively. The proportion of NALM-6 cells in peripheral blood and the proportion of CD19 CAR-T cells in T cells from the vein of the inner canthus were detected by flow cytometry. The overall survival of BAL B/c nude mice was observed. Results:The proliferation of mouse and humanized CD19 CAR-T cells were (68.50±0.93)% and (80.63±1.41)%, respectively ( t=20.353, P<0.001) after cultured in vitro for 24 hours, and were (91.38±1.41)% and (148.13±1.25)%, respectively ( t=85.364, P<0.001) after cultured for 48 hours. When the effect to target ratio was 1∶1, there was no difference between the humanized and murine CD19 CAR-T cell group after co-culture for 24 hours ( P=0.169), while the killing activity of humanized CD19 CAR-T cells against NALM-6 cells was higher than that of murine CD19 CAR-T cells ( P<0.01) after 48 hours of co-culture. When the effect to target ratio was 4∶1, the cytotoxicity of humanized CD19 CAR-T cells against NALM-6 cells was higher than that of murine CD19 CAR-T cells in co-culture for 24 and 48 hours ( P<0.01). On the seventh day of CD19 CAR-T cell therapy, the proportion of NALM-6 cells in the peripheral blood of BAL B/c nude mice decreased to the lowest level in the humanized CD19 CAR-T cell group and the murine CD19 CAR-T cell group. After 21 days, the proportion of NALM-6 cells in the murine CD19 CAR-T cell group was higher than that in the humanized CD19 CAR-T cell group ( P21 d=0.001, P28 d<0.001, P35 d<0.001). The proportion of humanized and murine CD19 CAR-T cells in the peripheral blood reached the peaks after 7 days of therapy, and the proportion of humanized CD19 CAR-T cells was higher than that of murine CAR-T cells ( P7 d=0.002). The CD19 CAR-T cells disappeared in the peripheral blood in the murine CD19 CAR-T cell group after 14 days of therapy, while in the humanized CD19 CAR-T cell group it disappeared after 21 days of therapy. The median survival of BAL B/c nude mice in the murine CD19 CAR-T cell group and the humanized CD19 CAR-T cell group was 42 days and 63 days, respectively ( χ2=15.382, P<0.001). Conclusions:High affinity humanized CD19 CAR-T cells have stronger proliferation, higher cytotoxicity and longer survival time compared with those of murine CD19 CAR-T cells. The results indicate that the clinical efficacy of humanized CD19 CAR-T cells would be better than that of murine CD19 CAR-T cells.

Objective:To compare the activity difference of the high affinity humanized CD19 chimeric antigen receptor (CAR)-T cells and murine CD19 CAR-T cells.Methods:Peripheral venous blood T cells from 8 healthy volunteers were collected and infected with humanized and murine CD19 CAR lentivirus. Human and murine CD19 CAR-T cells were prepared and cell proliferation was detected by cell counting kit-8 (CCK-8) method. The cytotoxicity of CD3 + T cells, humanized and murine CD19 CAR-T cells to NALM-6 cells was detected by lactate dehydrogenase assay. Thirty BAL B/c nude mice transplanted with NALM-6 cells were randomly divided into 3 groups with 10 mice in each group and injected humanized CD19 CAR-T cells, mouse CD19 CAR-T cells and control CD3 + T cell via tail vein, respectively. The proportion of NALM-6 cells in peripheral blood and the proportion of CD19 CAR-T cells in T cells from the vein of the inner canthus were detected by flow cytometry. The overall survival of BAL B/c nude mice was observed. Results:The proliferation of mouse and humanized CD19 CAR-T cells were (68.50±0.93)% and (80.63±1.41)%, respectively ( t=20.353, P<0.001) after cultured in vitro for 24 hours, and were (91.38±1.41)% and (148.13±1.25)%, respectively ( t=85.364, P<0.001) after cultured for 48 hours. When the effect to target ratio was 1∶1, there was no difference between the humanized and murine CD19 CAR-T cell group after co-culture for 24 hours ( P=0.169), while the killing activity of humanized CD19 CAR-T cells against NALM-6 cells was higher than that of murine CD19 CAR-T cells ( P<0.01) after 48 hours of co-culture. When the effect to target ratio was 4∶1, the cytotoxicity of humanized CD19 CAR-T cells against NALM-6 cells was higher than that of murine CD19 CAR-T cells in co-culture for 24 and 48 hours ( P<0.01). On the seventh day of CD19 CAR-T cell therapy, the proportion of NALM-6 cells in the peripheral blood of BAL B/c nude mice decreased to the lowest level in the humanized CD19 CAR-T cell group and the murine CD19 CAR-T cell group. After 21 days, the proportion of NALM-6 cells in the murine CD19 CAR-T cell group was higher than that in the humanized CD19 CAR-T cell group ( P21 d=0.001, P28 d<0.001, P35 d<0.001). The proportion of humanized and murine CD19 CAR-T cells in the peripheral blood reached the peaks after 7 days of therapy, and the proportion of humanized CD19 CAR-T cells was higher than that of murine CAR-T cells ( P7 d=0.002). The CD19 CAR-T cells disappeared in the peripheral blood in the murine CD19 CAR-T cell group after 14 days of therapy, while in the humanized CD19 CAR-T cell group it disappeared after 21 days of therapy. The median survival of BAL B/c nude mice in the murine CD19 CAR-T cell group and the humanized CD19 CAR-T cell group was 42 days and 63 days, respectively ( χ2=15.382, P<0.001). Conclusions:High affinity humanized CD19 CAR-T cells have stronger proliferation, higher cytotoxicity and longer survival time compared with those of murine CD19 CAR-T cells. The results indicate that the clinical efficacy of humanized CD19 CAR-T cells would be better than that of murine CD19 CAR-T cells.

Objective:To investigate the characteristics and cytotoxicity in vitro of the residual leukemia cells in the culture system that caused the accidental transfer of CD19 chimeric antigen receptor (CAR) into leukemia cells during the preparation of autologous CD19 CAR-T cells of relapsed/refractory B-cell acute lymphoblastic leukemia.Methods:①Peripheral blood mononuclear cells (PBMC) of 30 patients with relapsed/refractory B-cell acute lymphoblastic anemia (R/R B-ALL) who accepted CD19 CAR-T cell therapy and six healthy volunteers were collected. ②The residual leukemia cells were analyzed by flow cytometry in the system after the PBMCs of R/R B-ALL patients were sorted by CD3 magnetic beads. ③ CD3 + T cells from patients and healthy volunteers were transfected with CD19 CAR and CD22 CAR lentivirus to prepare CD19 CAR-T and CD22 CAR-T cells. ④The Nalm-6 cell line was resuscitated and the Nalm-6 cells with CD19 CAR lentivirus were transfected to prepare CD19 CAR-Nalm-6 cells. The patient's primary ALL cells were transfected with CD19 CAR lentivirus at the same time. ⑤The transfection rates were analyzed by flow cytometer, the cell proliferation was analyzed by the CCK-8 method, and the cell-killing activities were detected by the lactate dehydrogenase method. Results:① Among the 30 R/R B-ALL patients who received CD19 CAR-T cell therapy, two patients had 2.04% and 3.32% residual leukemia cells in CD3 + T cells. After 4 days in culture, the residual leukemia cells disappeared and could not be detected by a flow cytometer with prolonged cultivation in vitro. ② The proliferation of CD19 CAR-Nalm-6 cells was higher than that of the Nalm-6 cells. ③ The killing activity of the CD19 CAR-T cells on Nalm-6 cells was higher than that of the CD19 CAR-Nalm6 cells at a target ratio of 1∶1 on 24, 48, 72 h, respectively. The cytotoxicity of CD22 CAR-T cells on CD19 CAR-Nalm-6 cells was significantly higher than that of CD19 CAR-T cells. ④ The cytotoxicity of CD22 CAR-T alone on CD19 CAR-Nalm-6 cells was higher than that of CD19 CAR-T combined with CD22 CAR-T at the same target ratio. Conclusion:The residual leukemia cells in the culture system in the preparation of CD19 CAR-T cells may lead to the introduction of CD19 CAR into leukemia cells and results in the failure of the CD19 CAR-T cell therapy. Detecting the residual leukemia cells in the culture system via flow cytometry before transfection with CD19 CAR lentivirus is needed. Thus, CD22 CAR-T cell therapy could be used as one of the salvage treatments.

Objective:To observe the efficacy and safety of humanized anti-BCMA chimeric antigen receptor modified (BCMA CAR) -T cell therapy after disease progression with their murine BCMA CAR-T cell therapy in patients with relapsed/refractory multiple myeloma (MM) .Methods:Study participants underwent leukapheresis to collect T cells for BCMA CAR-T manufacturing. Patients were pretreated with intensive chemotherapy (fludarabine combined with cytarabine) before CAR-T therapy. Adverse events (AEs) , CAR DNA expansion, and cytokine were monitored. In vitro, transfection efficacy, specific cytotoxicity, and inflammatory response were detected when co-cultured with effector and target cells.Results:Patient (PT) 1 and 2 achieved complete remission (CR) and disease stability at 3 months post murine CAR-T therapy. However, 16 and 18 months later, they experienced progression of disease (PD) , and patient 1 presented with extramedullary disease at PD. Both of the patients received humanized CAR-T therapy and achieved partial remission (PR) and very good partial remission (VGPR) post humanized CAR-T therapy. PT1 achieved CR of the soft tissue masses at 4 months post humanized CAR-T therapy. Notably, the median peak of the BCMA CAR-T cells, copy of BCMA CAR gene, persistence of BCMA CAR-T, and the peak levels of IL-6, IL-8, IL-10, IFN-γ and TNF-α were higher in humanized CAR-T therapy than those in the murine CAR-T therapy. During the murine CAR-T therapy, both of the patients experienced grade 1 CRS and no ICANS. PT1 experienced grade 3 CRS and grade 2 ICANS during humanized CAR-T therapy, which were relieved by supportive care. Grade 2 CRS was observed for patient 2 during humanized CAR-T therapy. Humanized BCMA CAR-T cells showed a higher inflammatory response and in vitro cytotoxicity than that of murine BCMA CAR-T cells with effector/targets cells at 1∶1 over 48 hours ( P<0.001) . The proportions of residual cells in humanized BCMA CAR-T and murine CAR-T were (17.38±5.18) % vs (28.27±4.58) %, (13.25±1.62) % vs (22.77±1.77) % for PT1 and PT2, respectively. Conclusions:The humanized BCMA CAR-T cell therapy was efficient and safe for patients who experienced progression of disease after the murine CAR-T therapy, especially for patients with extramedullary disease.