Lactic acid bacteria (LAB) are promising candidates for live bacterial drug delivery systems owing to their safety, probiotic properties, and ability to colonize the intestinal tract. At present, most research focuses on engineering LAB as carriers for the delivery of therapeutic proteins. As model organisms equipped with a versatile set of genetic tools, LAB can be readily modified to target various diseases and yield significant therapeutic effects. LAB-based carriers offer multiple advantages, including non-invasive delivery, ease of genetic manipulation, and suitability for large-scale production. Consequently, the use of LAB as recombinant protein expression vectors has attracted extensive research interest worldwide. The foundational principles, strategies for enhancing bioavailability, genetic engineering approaches, and the current research and application status of LAB-based drug delivery systems were summarized in this paper.

Objective To investigate the effects of scribble planar cell polarity protein(SCRIB)on proliferation,apoptosis,and autophagy of glioblastoma(GBM)and elucidate its potential underlying mechanisms.Methods The expression level of SCRIB in GBM tissue was queried through the Biomarker Exploration of Solid Tumors(BEST)database.Lentivirus-mediated shRNA interference was employed to downregulate SCRIB expression in human glioblastoma cell lines U87 and U251,which were divided into negative control group(mock)and SCRIB shRNA interference groups(kd1 and kd2).SCRIB expression levels were detected using Western blotting(WB)and quantitative polymerase chain reaction(qPCR).EdU incorporation and cell apoptosis rates were detected by flow cytometry(FCM).CCK-8 assay was used to detect the proliferation vitality of U87 and U251 cells,and WB was used to detect the expression of proliferation-related proteins.Immunofluorescence(IF)staining was conducted to detect the expression of autophagy-related proteins LC3 and p62,followed by quantitative analysis across multiple fields.WB was also used to detect the expression levels of LC3,p62,and proteins in the JAK-STAT3 signaling pathway.Results Compared with that of normal tissues,SCRIB mRNA expression level was significantly upregulated in GBM tissues(P＜0.05).FCM results showed that EdU incorporation rates were significantly reduced(P＜0.001)while cell apoptosis rates were markedly increased(P＜0.001)in U87 and U251 cells with SCRIB knockdown.CCK-8 results indicated that compared with the mock group,the proliferation vitality of U87 and U251 cells in the SCRIB knockdown group was significantly downregulated(P＜0.001).IF staining showed that LC3 fluorescence aggregation was significantly enhanced(P＜0.001),while p62 fluorescence aggregation was significantly reduced(P＜0.001)in the SCRIB knockdown group.WB results showed that compared with the mock group,the protein expression levels of p27,LC3,p-JAK2 and p-STAT3 were upregulated,while C-Myc,Cyclin D1,MCM,PCNA and p62 were downregulated,with statistically significant differences(P＜0.05).Conclusion Downregulation of SCRIB may induce autophagy and apoptosis in glioblastoma cells by inhibiting the JAK-STAT3 signaling pathway,thereby suppressing cell proliferation.

Objective To investigate the ameliorative effects and mechanisms of six types of tea(green tea,cyan tea,red tea,white tea,black tea and yellow tea)on metabolic disorders in obesity mice induced by high-fat diet(HFD).Methods Four-week-old male C57BL/6J mice were randomly divided into 8 groups with 7 mice per group.An HFD-induced obese mouse model was established,and the mice in control group maintained on standard diet followed by intragastric administration of different teas for 5 weeks.The body weight,liver weight ratio,fasting blood glucose,and lipid profile of the mice were measured to assess glucose and lipid metabolism.Serum inflammatory factors including IL-6,tumor necrosis factor-alpha(TNF-α)and oxidative stress markers[malondialdehyde(MDA)and superoxide dismutase(SOD)were measured.Additionally,liver histopathology and the expression of key glycolipid metabolism-related genes,adenosine monophosphate-activated protein kinase(AMPK)and carnitine palmitoyltransferase 1(CPT-1),were analyzed to explore underlying mechanisms.Results Cyan tea significantly suppressed weight gain,demonstrating superior weight control.White tea markedly reduced fasting blood glucose levels and decreased the area under the curve of oral glucose tolerance test(OGTT)and insulin tolerance test(ITT),indicating synergistic improvements in glucose metabolism and insulin sensitivity.Yellow tea exhibited exceptional anti-inflammatory and antioxidant effects,reducing hepatic IL-6 and MDA while enhancing SOD activity.Green tea activated the lipid oxidation pathway by upregulating AMPK/CPT-1 expression.All kinds of tea significantly attenuated hepatic lipid droplet accumulation.Conclusion All six types of tea alleviated metabolic disorders by reducing hepatic fat content in obesity mice.However,different types of tea exert their unique effects on improving metabolic disorders through differential mechanisms such as glucose metabolism regulation,lipid oxidation,and anti-inflammatory and antioxidant actions.

Objective:To document any effect of repeated high-frequency repetitive transcranial magnetic stimulation (rTMS) using double-cone coils on the lower limb motor function of stroke survivors.Methods:A total of 40 stroke survivors were randomly divided into an rTMS group and a sham stimulation group, each of 20. The rTMS group received rTMS at 10Hz with a double-cone coil, while a coil that produced sound but no magnetic stimulation was used with the sham group. The treatments were administered daily, five times a week, for three weeks. Before as well as after 1, 2 and 3 weeks of treatment, lower limb motor function, balance, and the root mean square (RMS) and median frequency (MF) of the rectus femoris and tibialis anterior muscles were evaluated using the Fugl-Meyer lower extremity assessment (FMA-LE), the Berg Balance Scale (BBS), the Modified Barthel Index (MBI), Brunnstrom staging, the TecnoBody balance assessment system, and surface electromyography.Results:Compared with the sham stimulation group, the BBS score of the rTMS group was significantly higher after 2 weeks of treatment, and both the FMA-LE and BBS scores were significantly higher after 3 weeks. The average Brunnstrom stage in the sham group had increased significantly after 3 weeks, but in the rTMS group it had increased after 2 weeks. By 3 weeks there were no significant differences between the two groups. In terms of movement control, the average motion ellipse area in the rTMS group was significantly smaller than among the sham group after 2 weeks, and after 3 weeks the average motion trajectory length was significantly shorter than in the sham group. The average RMS of the rectus femoris in the rTMS group was significantly higher than the sham group′s average after 3 weeks of treatment, indicating improved muscle activation.Conclusions:High-frequency rTMS using a conical coil can effectively improve the lower limb motor function and balance ability of stroke survivors, demonstrating promising clinical application potential.

To investigate the effects of zearalenone(ZEA)on the proliferation and apoptosis of sika deer antler chondrocytes,the chondrocytes were isolated and cultured in vitro and treated with 50μmol/L ZEA for 24 h.Flow cytometry was used to assess cell proliferation,cell cycle,apoptosis,mitochondrial membrane potential,and intracellular levels of reactive oxygen species(ROS).The expression changes of hypertrophic cartilage cell marker genes Col X,Runx2,Alpl,and apoptosis-related genes Casp-3,Bax,Bcl-2 were measured using quantitative PCR.Additionally,glutathione reductase(GR)activity and the levels of the oxidative stress marker malondialdehyde(MDA)were determined.The results showed that after 24 h of ZEA treatment,cell proliferation was sig-nificantly inhibited,with an increase in the number of cells in the G0/G1 phase and a decrease in the S phase.The expression levels of hypertrophic chondrocyte marker genes Col X,Runx2 and Al-pl were significantly increased.Apoptosis rate was significantly increased,with elevated expression of pro-apoptotic genes Casp-3,Bax and reduced expression of the anti-apoptotic gene Bcl-2.The content of MDA in the antler chondrocytes increased,ROS levels rose,and GR activity decreased.The mitochondrial membrane potential reduced.The results suggested that ZEA could inhibit the proliferation of antler chondrocytes and promote the apoptosis by regulating cellular oxidative stress responses and the expression of apoptosis-related genes.

Objective:To document any effect of repeated high-frequency repetitive transcranial magnetic stimulation (rTMS) using double-cone coils on the lower limb motor function of stroke survivors.Methods:A total of 40 stroke survivors were randomly divided into an rTMS group and a sham stimulation group, each of 20. The rTMS group received rTMS at 10Hz with a double-cone coil, while a coil that produced sound but no magnetic stimulation was used with the sham group. The treatments were administered daily, five times a week, for three weeks. Before as well as after 1, 2 and 3 weeks of treatment, lower limb motor function, balance, and the root mean square (RMS) and median frequency (MF) of the rectus femoris and tibialis anterior muscles were evaluated using the Fugl-Meyer lower extremity assessment (FMA-LE), the Berg Balance Scale (BBS), the Modified Barthel Index (MBI), Brunnstrom staging, the TecnoBody balance assessment system, and surface electromyography.Results:Compared with the sham stimulation group, the BBS score of the rTMS group was significantly higher after 2 weeks of treatment, and both the FMA-LE and BBS scores were significantly higher after 3 weeks. The average Brunnstrom stage in the sham group had increased significantly after 3 weeks, but in the rTMS group it had increased after 2 weeks. By 3 weeks there were no significant differences between the two groups. In terms of movement control, the average motion ellipse area in the rTMS group was significantly smaller than among the sham group after 2 weeks, and after 3 weeks the average motion trajectory length was significantly shorter than in the sham group. The average RMS of the rectus femoris in the rTMS group was significantly higher than the sham group′s average after 3 weeks of treatment, indicating improved muscle activation.Conclusions:High-frequency rTMS using a conical coil can effectively improve the lower limb motor function and balance ability of stroke survivors, demonstrating promising clinical application potential.

To investigate the effects of zearalenone(ZEA)on the proliferation and apoptosis of sika deer antler chondrocytes,the chondrocytes were isolated and cultured in vitro and treated with 50μmol/L ZEA for 24 h.Flow cytometry was used to assess cell proliferation,cell cycle,apoptosis,mitochondrial membrane potential,and intracellular levels of reactive oxygen species(ROS).The expression changes of hypertrophic cartilage cell marker genes Col X,Runx2,Alpl,and apoptosis-related genes Casp-3,Bax,Bcl-2 were measured using quantitative PCR.Additionally,glutathione reductase(GR)activity and the levels of the oxidative stress marker malondialdehyde(MDA)were determined.The results showed that after 24 h of ZEA treatment,cell proliferation was sig-nificantly inhibited,with an increase in the number of cells in the G0/G1 phase and a decrease in the S phase.The expression levels of hypertrophic chondrocyte marker genes Col X,Runx2 and Al-pl were significantly increased.Apoptosis rate was significantly increased,with elevated expression of pro-apoptotic genes Casp-3,Bax and reduced expression of the anti-apoptotic gene Bcl-2.The content of MDA in the antler chondrocytes increased,ROS levels rose,and GR activity decreased.The mitochondrial membrane potential reduced.The results suggested that ZEA could inhibit the proliferation of antler chondrocytes and promote the apoptosis by regulating cellular oxidative stress responses and the expression of apoptosis-related genes.

Objective:To investigate the association between adiponectin-related copy number variation (CNV) region (CNVR) and gestational diabetes mellitus (GDM).Methods:Pregnant women who had prenatal screening in Anqing Municipal Hospital, Anhui Province, from February 2018 to December 2020 were surveyed for baseline information collection, and blood samples were collected. The outcome information was obtained by post pregnancy follow-up. Latex-enhanced immunoturbidimetry and ASA-CHIA chip were used to detect serum adiponectin levels and CNV of pregnant women, respectively. After genotyping, CNV data were processed with software PennCNV 1.0.5 following standard quality control procedure. CNVR were identified and integrated by using software R 4.3.3. Then the associations between CNVR and adiponectin was evaluated, and gene annotation and over-representation analysis were conducted. The log-binomial regression model was used to adjust relevant covariates and analyze the association between adiponectin-related CNVR and GDM.Results:The detection rate of GDM was 9.54% (176/1 845) in the pregnant women. The genotyping information of 1 840 people (99.73%) passed quality evaluation. A total of 33 878 CNVs were identified, and 1 449 CNVRs were obtained after integration. After the false discovery rate method correction, CNVR_132 (CHR2: 47611743-47635062), CNVR_254 (CHR3: 10182703-10183872), CNVR_691 (CHR7: 150637053-150834539) and CNVR_1101 (CHR14: 104248431-104830620) were correlated with adiponectin levels (all P<0.05). Over- representation analysis showed that the molecular function of ribonucleotide binding [Gene Ontology (GO): 0032553] was significantly enriched based on the GO database. The log-binomial regression model, adjusting age, pre-pregnancy BMI, history of miscarriage, smoking history, and family history of diabetes, indicated that CNVR_132 (CHR2: 47611743-47635062) and CNVR_1101 (CHR14: 104248431-104830620) were not statistically associated with the risk for GDM (both P>0.05). However, CNVR_254 (CHR3: 10182703-10183872, a RR=1.83, 95% CI: 1.15-2.91) and CNVR_691 (CHR7: 150637053-150834539, a RR=1.73, 95% CI: 1.23-2.43) might be associated with an increased risk for GDM (all P<0.05). Conclusion:Adiponectin-related CNVR_254 (CHR3: 10182703-10183872) and CNVR_691 (CHR7: 150637053-150834539) might be risk factors for the incidence of GDM.

Objective To investigate the effects of scribble planar cell polarity protein(SCRIB)on proliferation,apoptosis,and autophagy of glioblastoma(GBM)and elucidate its potential underlying mechanisms.Methods The expression level of SCRIB in GBM tissue was queried through the Biomarker Exploration of Solid Tumors(BEST)database.Lentivirus-mediated shRNA interference was employed to downregulate SCRIB expression in human glioblastoma cell lines U87 and U251,which were divided into negative control group(mock)and SCRIB shRNA interference groups(kd1 and kd2).SCRIB expression levels were detected using Western blotting(WB)and quantitative polymerase chain reaction(qPCR).EdU incorporation and cell apoptosis rates were detected by flow cytometry(FCM).CCK-8 assay was used to detect the proliferation vitality of U87 and U251 cells,and WB was used to detect the expression of proliferation-related proteins.Immunofluorescence(IF)staining was conducted to detect the expression of autophagy-related proteins LC3 and p62,followed by quantitative analysis across multiple fields.WB was also used to detect the expression levels of LC3,p62,and proteins in the JAK-STAT3 signaling pathway.Results Compared with that of normal tissues,SCRIB mRNA expression level was significantly upregulated in GBM tissues(P＜0.05).FCM results showed that EdU incorporation rates were significantly reduced(P＜0.001)while cell apoptosis rates were markedly increased(P＜0.001)in U87 and U251 cells with SCRIB knockdown.CCK-8 results indicated that compared with the mock group,the proliferation vitality of U87 and U251 cells in the SCRIB knockdown group was significantly downregulated(P＜0.001).IF staining showed that LC3 fluorescence aggregation was significantly enhanced(P＜0.001),while p62 fluorescence aggregation was significantly reduced(P＜0.001)in the SCRIB knockdown group.WB results showed that compared with the mock group,the protein expression levels of p27,LC3,p-JAK2 and p-STAT3 were upregulated,while C-Myc,Cyclin D1,MCM,PCNA and p62 were downregulated,with statistically significant differences(P＜0.05).Conclusion Downregulation of SCRIB may induce autophagy and apoptosis in glioblastoma cells by inhibiting the JAK-STAT3 signaling pathway,thereby suppressing cell proliferation.

Objective:To quantitatively analyze right ventricular reverse remodeling in patients with severe tricuspid regurgitation after transcatheter tricuspid edge-to-edge repair (T-TEER) by two-dimensional speckle tracking echocardiography, and to preliminarily evaluate the clinical efficacy of this procedure.Methods:This study was a prospective single-center cohort study. Patients diagnosed with severe tricuspid regurgitation at the Xiamen Cardiovascular Hospital Xiamen University from March 2021 to June 2023 were enrolled. All patients underwent transthoracic echocardiography and transesophageal three-dimensional echocardiography before T-TEER, and transthoracic echocardiography at 30 days, 6 months, and 9 months after T-TEER. The primary endpoint was major adverse cardiovascular and cerebrovascular events, including death, stroke, myocardial infarction, reoperation, arrhythmia, and conduction block. Other clinical evaluation indicators included New York Heart Association (NYHA) functional classification and tricuspid regurgitation grade.Results:A total of 34 patients were enrolled, aged (67.9±9.3) years, and 71% (24/34) were female. The median follow-up duration was 9 months. All patients achieved a reduction of tricuspid regurgitation by ≥2 grades at 9 months after T-TEER, with 79% (27/34) of them having mild to moderate tricuspid regurgitation. Transthoracic echocardiography at 9 months after T-TEER showed that the vena contracta width of tricuspid regurgitation ((5.42±2.33) mm vs. (11.54±4.05) mm, P<0.001), effective regurgitant orifice area ((0.24±0.09) cm2 vs. (0.52±0.14) cm2, P<0.001), regurgitant jet area ((7.95±4.02) cm2 vs. (13.93±6.10) cm2, P<0.001), inferior vena cava diameter ((19.38±2.63) mm vs. (23.56±3.31) mm, P<0.001), right ventricular end-diastolic diameter ((28.03±6.26) mm vs. (33.21±8.24) mm, P=0.001), and tricuspid annular diameter ((36.47±4.40) mm vs. (41.44±7.08) mm, P<0.001) were all reduced compared with baseline; while the tricuspid annular plane systolic excursion ((18.08±5.25) mm vs. (14.91±3.42) mm, P=0.005) and right ventricular fractional area change ((37.61±7.52)% vs. (30.79±9.06)%, P=0.004) were both increased compared with baseline. At 9 months after T-TEER, all patients had a NYHA functional classification of grade Ⅰ or Ⅱ, and no major adverse cardiovascular and cerebrovascular event occurred during the follow-up period. Conclusion:It is preliminarily confirmed that T-TEER is safe and effective in the treatment of severe tricuspid regurgitation, with significant right ventricular reverse remodeling observed in patients at 9 months after T-TEER.