Objective To investigate the effect of FUNDC1 tyrosine phosphorylation site mutations on mitophagy in H9c2 myocardial cells by constructing tyrosine site mutant plasmids (Y11 and Y18) of the FUN14 domain-containing protein 1 (FUNDC1). Methods The mutant plasmids constructed by whole-gene synthesis were transfected into rat myocardial H9c2 cells and divided into five groups: empty plasmid group, FUNDC1 overexpression group, Y11 mutant group, Y18 mutant group, and Y11 combined with Y18 mutant group. The viability of H9c2 cells was assessed using the CCK-8 assay. Additionally, tetramethylrhodamine ethyl ester (TMRE) staining was utilized to detect mitochondrial membrane potential. The protein expression levels of FUNDC1, translocase of the outer mitochondrial membrane 20 (TOM20), and cytochrome c oxidase IV (COX IV) were detected by Western blot analysis. Confocal microscopy was used to evaluate transfection efficiency as well as the co-localization of mitochondria and lysosomes. Results The FUNDC1 overexpression, Y11, Y18, and Y11 combined with Y18 mutant plasmids were successfully constructed. After plasmid transfection, widespread GFP fluorescence expression was observed under confocal microscopy. Compared with the empty plasmid group, FUNDC1 protein expression levels were significantly increased in the FUNDC1 overexpression group, Y11 mutation group, Y18 mutation group, and Y11 combined with Y18 mutation group, while cell viability and mitochondrial membrane potential showed no significant changes. Compared to the empty plasmid group, cells transfected with Y18 and Y11 combined with Y18 mutant plasmids showed increased TOM20 and COX IV expression levels and decreased mitochondrial-lysosomal co-localization. Conclusion Transfection with FUNDC1 Y18 or Y11 combined with Y18 mutant plasmids inhibited mitophagy in H9c2 myocardial cells.
Animals ; Rats ; Mitophagy/genetics* ; Myocytes, Cardiac/cytology* ; Mitochondrial Proteins/metabolism* ; Mutation ; Phosphorylation ; Tyrosine/genetics* ; Cell Line ; Membrane Proteins/metabolism* ; Membrane Potential, Mitochondrial

Objective:To investigate GPRC5D expression on myeloma cells in newly diagnosed multiple myeloma (NDMM) patients and evaluate its prognostic significance.Methods:This study retrospectively analyzed the clinical data of 65 patients with NDMM treated at the Affiliated Hospital of Xuzhou Medical University from April 2023 to April 2024. The expression of GPRC5D on the surface of myeloma cells in all patients was detected with flow cytometry before induction therapy, and patients were stratified into high and low GPRC5D expression groups based on the median GPRC5D positivity rate. Clinical characteristics, immune status, treatment response after induction therapy, and prognosis were compared between the two groups.Results:The median positive rate of GPRC5D in the plasma cells of 65 patients with NDMM was 32.68%. Based on this threshold, patients were categorized into the high (33 cases, GPRC5D positive rate ≥ 32.68%) and low (32 cases, GPRC5D positive rate <32.68%) GPRC5D expression groups. Compared with the low GPRC5D expression group, the high GPRC5D expression group demonstrated a higher proportion of 1q21 gain (78.8% vs 43.8%, P=0.004), a higher incidence of immunoparesis involving ≥2 uninvolved immunoglobulins (87.9% vs 62.5%, P=0.018), and severe immunoparesis (59.4% vs 33.3%, P=0.046). Further, CD16 +CD56 + cell levels were lower in the high GPRC5D expression group [ (16.60±8.70) % vs (27.78±15.78) %, P=0.005]. No significant difference was observed in the overall response rate between the high and low GPRC5D expression groups (78.8% vs 93.8%, P=0.165). However, the high GPRC5D expression group exhibited a significantly lower rate of achieving very good partial remission or better (42.4% vs 78.2%, P=0.003) and a lower MRD negativity rate (30.0% vs 68.8%, P=0.002). Compared with the low GPRC5D expression group, patients with high expression demonstrated a significantly shorter median progression-free survival (11.2 months vs not reached, P=0.002), whereas the median overall survival was not reached in either group, with no statistically significant difference ( P=0.069) . Conclusions:The GPRC5D positivity rate in the plasma cells of patients with NDMM is associated with 1q21 gain and immune status. High GPRC5D expression at diagnosis may predict poor response to induction therapy and an unfavorable prognosis.

Objective: To compare and analyze the antibacterial effects of platelets against five common clinical pathogenic bacteria including MRSA, SE, SA, E. coli, and CRKP, and to preliminarily explore the role of DCD sensitivity in the observed variations of antibacterial effects. Methods: The same number of platelets were used to establish co-culture systems of platelets and platelet lysates with the five pathogenic bacteria. The antibacterial effects of platelets and platelet lysates on the five pathogenic bacteria were evaluated by observing the turbidity of the bacterial solution, measuring the OD value of the bacterial solution and counting the colonies. The supernatant protein of platelets co-cultured with MRSA was collected for quantitative proteomics analysis to explore the important antibacterial proteins of platelets. The content of DCD in the supernatant after co-culture of platelets and platelet lysates with the five pathogenic bacteria was detected by ELISA to preliminarily analyze the reasons for the different antibacterial effects of platelets on the five pathogenic bacteria. Results: Compared with the control group of MRSA, SA, and SE, the turbidity of the bacterial solution decreased after co-culture of platelets and platelet lysates with MRSA, SA, and SE for 12 h, and the OD value and colony count were significantly reduced (P<0.05). The turbidity of the bacterial solution did not change significantly after co-culture of platelets and platelet lysates with E. coli for 24 h, but the OD value decreased (P<0.05), and the colony count decreased to 10 CFU/mL but the difference was not statistically significant (P>0.05). Compared with the control group of CRKP, the turbidity, OD value, and colony count of the bacterial solution did not change significantly after co-culture of platelets and platelet lysates with CRKP (P>0.05). Proteomics results showed that after co-culture with MRSA, important proteins related to platelet activation, including collagen, fibrinogen, von Willebrand factor, integrin αIIbβ3, platelet glycoprotein V and IV were significantly up-regulated. ELISA results showed that after co-culture with the five pathogenic bacteria, platelets could secrete a large amount of DCD, with the content around 3 μg/mL. Conclusion: The antibacterial effect of platelets on Gram-positive bacteria MRSA, SA, and SE is better than that on Gram-negative bacteria E. coli and CRKP, and platelets have the best antibacterial effect on MRSA. The differences in antibacterial effects of platelets on the five pathogenic bacteria may be related to the sensitivity of DCD antibacterial peptides to the five pathogenic bacteria.

Objective To investigate the correlation and clinical prognostic value between the expression of polypeptide N-acety lgalactosaminyltransferases 1(GALNT1),lysosomal adaptor MTOR activator 5(LAMTOR5),and epithelial mesenchymal transition(EMT)in gastric cancer.Methods 102 gastric cancer patients admitted to Nantong Tumor Hospital from January 2018 to January 2020 were selected.Immunohistochemistry(IHC)and real-time fluorescence quantitative PCR(RT-PCR)were used to detect the expression of GALNT1,LAMTOR5 mRNA and protein,as well as the expression of EMT indicators Snail mRNA,N-cad mRNA and Vimentin mRNA in cancer and adjacent tissues.Pearson correlation analysis was performed to investigate the correlation between markers.Kaplan Meier curve analysis was used to investigate the relationship between GALNT1 mRNA,LAMTOR5 mRNA and the prognosis of gastric cancer patients.COX regression analysis was used to identify factors that affect the prognosis of gastric cancer patients.Results GALNT1 mRNA(3.38±0.29)and LAMTOR5 mRNA(3.17±0.26)in gastric cancer tissue were higher than those in adjacent tissues(0.72±0.16,0.65±0.13),and the differences were statistically significant(t=81.111,87.553,all P<0.001).The positive rates of GALNT1(92.16%),LAMTOR5(90.20%)proteins in gastric cancer tissues were higher than those in adjacent tissues(7.84%,11.76%),and the differences were statistically significant(χ2=145.020,125.538,P<0.001).The expression of GALNT1 mRNA and LAMTOR5 mRNA in gastric cancer was positively correlated(r=0.757,P<0.001).The expression of GALNT1 mRNA and LAMTOR5 mRNA in gastric cancer was significantly positively correlated with Snail mRNA,N-cad mRNA and Vimentin mRNA expression(r=0.654,0.712,0.689;0.706,0.645,0.723,all P<0.001).The expression of GALNT1 mRNA and LAMTOR5 mRNA was correlated with TNM stage and lymph node metastasis in gastric cancer patients.The expression of GALNT1 mRNA and LAMTOR5 mRNA in cancer tissues with TNM stage III and lymph node metastasis was higher than that in TNM stage I～II and no lymph node metastasis,and the differences were statistically significant(χ2=29.417～290.104,all P<0.001).The 3-year overall survival rate of the high expression group of GALNT1 mRNA was 40.00%(20/50),which was lower than 76.92%(40/52)of the low expression group,the 3-year overall survival rate of the high expression group of LAMTOR5 mRNA was 38.78%(19/49),which was lower than the 77.36%(41/53)of the low expression group,and the differences were statistically significant(Log-Rank χ2=7.327,5.197,P=0.009,0.023).TNM stage III,lymph node metastasis,GALNT1 mRNA was highly expressed,and high LAMTOR5 mRNA was highly expressed were risk factors affecting the prognosis of gastric cancer(Wald χ2=6.409～16.805,all P<0.001).Conclusion The expression of GALNT1 and LAMTOR5 are elevated in gastric cancer tissues.Both of them are related to EMT indicators and adverse clinicopathological features.They are biomarkers for evaluating the prognosis of gastric cancer patients.

Objective To investigate the correlation and clinical prognostic value between the expression of polypeptide N-acety lgalactosaminyltransferases 1(GALNT1),lysosomal adaptor MTOR activator 5(LAMTOR5),and epithelial mesenchymal transition(EMT)in gastric cancer.Methods 102 gastric cancer patients admitted to Nantong Tumor Hospital from January 2018 to January 2020 were selected.Immunohistochemistry(IHC)and real-time fluorescence quantitative PCR(RT-PCR)were used to detect the expression of GALNT1,LAMTOR5 mRNA and protein,as well as the expression of EMT indicators Snail mRNA,N-cad mRNA and Vimentin mRNA in cancer and adjacent tissues.Pearson correlation analysis was performed to investigate the correlation between markers.Kaplan Meier curve analysis was used to investigate the relationship between GALNT1 mRNA,LAMTOR5 mRNA and the prognosis of gastric cancer patients.COX regression analysis was used to identify factors that affect the prognosis of gastric cancer patients.Results GALNT1 mRNA(3.38±0.29)and LAMTOR5 mRNA(3.17±0.26)in gastric cancer tissue were higher than those in adjacent tissues(0.72±0.16,0.65±0.13),and the differences were statistically significant(t=81.111,87.553,all P<0.001).The positive rates of GALNT1(92.16%),LAMTOR5(90.20%)proteins in gastric cancer tissues were higher than those in adjacent tissues(7.84%,11.76%),and the differences were statistically significant(χ2=145.020,125.538,P<0.001).The expression of GALNT1 mRNA and LAMTOR5 mRNA in gastric cancer was positively correlated(r=0.757,P<0.001).The expression of GALNT1 mRNA and LAMTOR5 mRNA in gastric cancer was significantly positively correlated with Snail mRNA,N-cad mRNA and Vimentin mRNA expression(r=0.654,0.712,0.689;0.706,0.645,0.723,all P<0.001).The expression of GALNT1 mRNA and LAMTOR5 mRNA was correlated with TNM stage and lymph node metastasis in gastric cancer patients.The expression of GALNT1 mRNA and LAMTOR5 mRNA in cancer tissues with TNM stage III and lymph node metastasis was higher than that in TNM stage I～II and no lymph node metastasis,and the differences were statistically significant(χ2=29.417～290.104,all P<0.001).The 3-year overall survival rate of the high expression group of GALNT1 mRNA was 40.00%(20/50),which was lower than 76.92%(40/52)of the low expression group,the 3-year overall survival rate of the high expression group of LAMTOR5 mRNA was 38.78%(19/49),which was lower than the 77.36%(41/53)of the low expression group,and the differences were statistically significant(Log-Rank χ2=7.327,5.197,P=0.009,0.023).TNM stage III,lymph node metastasis,GALNT1 mRNA was highly expressed,and high LAMTOR5 mRNA was highly expressed were risk factors affecting the prognosis of gastric cancer(Wald χ2=6.409～16.805,all P<0.001).Conclusion The expression of GALNT1 and LAMTOR5 are elevated in gastric cancer tissues.Both of them are related to EMT indicators and adverse clinicopathological features.They are biomarkers for evaluating the prognosis of gastric cancer patients.

Objective:To investigate GPRC5D expression on myeloma cells in newly diagnosed multiple myeloma (NDMM) patients and evaluate its prognostic significance.Methods:This study retrospectively analyzed the clinical data of 65 patients with NDMM treated at the Affiliated Hospital of Xuzhou Medical University from April 2023 to April 2024. The expression of GPRC5D on the surface of myeloma cells in all patients was detected with flow cytometry before induction therapy, and patients were stratified into high and low GPRC5D expression groups based on the median GPRC5D positivity rate. Clinical characteristics, immune status, treatment response after induction therapy, and prognosis were compared between the two groups.Results:The median positive rate of GPRC5D in the plasma cells of 65 patients with NDMM was 32.68%. Based on this threshold, patients were categorized into the high (33 cases, GPRC5D positive rate ≥ 32.68%) and low (32 cases, GPRC5D positive rate <32.68%) GPRC5D expression groups. Compared with the low GPRC5D expression group, the high GPRC5D expression group demonstrated a higher proportion of 1q21 gain (78.8% vs 43.8%, P=0.004), a higher incidence of immunoparesis involving ≥2 uninvolved immunoglobulins (87.9% vs 62.5%, P=0.018), and severe immunoparesis (59.4% vs 33.3%, P=0.046). Further, CD16 +CD56 + cell levels were lower in the high GPRC5D expression group [ (16.60±8.70) % vs (27.78±15.78) %, P=0.005]. No significant difference was observed in the overall response rate between the high and low GPRC5D expression groups (78.8% vs 93.8%, P=0.165). However, the high GPRC5D expression group exhibited a significantly lower rate of achieving very good partial remission or better (42.4% vs 78.2%, P=0.003) and a lower MRD negativity rate (30.0% vs 68.8%, P=0.002). Compared with the low GPRC5D expression group, patients with high expression demonstrated a significantly shorter median progression-free survival (11.2 months vs not reached, P=0.002), whereas the median overall survival was not reached in either group, with no statistically significant difference ( P=0.069) . Conclusions:The GPRC5D positivity rate in the plasma cells of patients with NDMM is associated with 1q21 gain and immune status. High GPRC5D expression at diagnosis may predict poor response to induction therapy and an unfavorable prognosis.

Biological hydroxyapatite(BHA)is widely used in the treatment of clinical bone defects due to its good biocompatibility and osteoconductivity.The clinical application of mateiral size is based on the principle of bone defect area adaptation,which contributes to diversity of BHA sizes.However,different sizes correspond to different hierarchi-cal levels of bone biomimicry.As the size changes,the bone biomimicry hierarchy evolves accordingly and influences the process of bone repair and regeneration through osteo-coagulo-immunomodulation,leading to unstable bone graft outcomes.Therefore,this paper reviews the size effect of clinical BHA,analyzes the multilevel structure of natural bone,proposes the evolution of bone biomimetic hierarchy triggered by the size of BHA,and further analyzes the size-media-ted osteo-coagulo-immunomodulation.Based on the hierarchical levels of bone and its osteo-coagulo-immunomodula-tion effect,we provide a new understanding of the biolog-ical principle of the size effect of biomaterials and a theo-retical basis for the basic research and clinical application of different size BHA materials.

Objective:To analyze TCM tongue diagnosis related research on coronary heart disease (CHD) at home and abroad using the method of bibliometrics; To provide reference for further research on clinical diagnosis and treatment of CHD.Methods:Research literature about TCM tongue diagnosis for CHD was retrieved from CNKI, VIP, CBM, Wanfang Data, PubMed, Embase, Cochrane Library and Web of Science from the establishment of the databases to December 31st, 2022. The key features of literature related to tongue diagnosis from CHD, including publication volume, region, institution, authors, and research hotspots, were analyzed using BICOMBS2.0, CiteSpace 6.1.R6, and Excel 2019 software.Results:Totally 364 Chinese articles and 5 English articles were included. Liaoning University of Traditional Chinese Medicine was the institution that published the most relevant research in Chinese, and Shanghai University of Traditional Chinese Medicine was the institution that published the most relevant research in English. Professor Wang Yiqin of Shanghai University of Traditional Chinese Medicine was the author with the largest number of publications in both Chinese and English, and the authors with the largest number of publications on Chinese research worked closely cooperate with each other. Chinese research focused on syndrome characteristics of CHD, objectification of tongue diagnosis and data mining based on the clinical experience of renowned TCM practitioners; the English research topic included modern data collection and analysis and expert consensus.Conclusions:The research team of tongue diagnosis for CHD in China has begun to take shape. There are few English articles on tongue diagnosis for CHD, and the development level is relatively lagging behind. In the future, different institutions and researchers need to strengthen international cooperation to enhance the international influence of TCM diagnostics.

Objective To clarify the distribution of traditional Chinese medicine(TCM)pattern and its associated risk factors after percutaneous coronary intervention(PCI),and evaluate the re-porting quality of existing studies to guide future research standardization. Methods English databases including PubMed,Cochrane Library,and Web of Science,as well as Chinese databases including China National Knowledge Infrastructure(CNKI),China Scientific Journal Database(VIP),and Wanfang Database were searched to retrieve papers about PCI.The time span for the paper retrieval was set from the foundation of the databases to October 1,2023.Statistical analyses were performed using Stata 12 and Python(V 3.9).The Strengthening the Reporting of Observational Studies in Epidemiology(STROBE)statement was used to assess the reporting quality of included studies. Results Overall,1 356 articles were selected,and 40 cross-sectional studies were included with 10 270 participants.The most common TCM patterns before,one to two weeks after,and six months to one year after PCI was Qi stagnation and blood stasis(n=261,36.45%),inter-twined phlegm and blood stasis(n=109,27.18%),and Qi deficiency and blood stasis(n=645,37.03%),respectively.Smoking[odds ratio(OR)=1.15,95%confidence interval(CI)(0.83-1.47),I2=24.7%,P=0.257],pattern of congealing cold and Qi stagnation[OR=4.62,95%CI(1.37-7.86),I2=61.6%,P=0.074],and low-density lipoprotein(LDL)[OR=1.38,95%CI(0.92-1.85),I2=12.2%,P=0.286]were risk factors for restenosis.Hypertension[OR=7.26,95%CI(3.54-14.88),I2=91.6%,P=0.001],and overweight[i.e.,body mass index(BMI)>23][OR=1.20,95%CI(1.07-1.35),I2=85.3%,P=0.009]were significant risk factors of concomi-tant anxiety. Conclusion This systematic review and meta-analysis revealed that patients with different TCM pattern types have distinct characteristics and risk factors after PCI.More high-quality studies are warranted to provide supportive evidence for future research and clinical practice.

[Objective]Depression is a common mental illness with a profound impact on physical health.Depression has been associated with a higher risk of bacterial infection;however,whether this relationship is causal and how depression affects infection remains unclear.Therefore,we aimed to investigate the effects of depressive phenotype in infected mice by constructing a chronic unpredictable mild stress(CUMS)model.[Methods]Mice were induced with CUMS for 4 weeks.The depressive phenotype was evaluated using behavioral tests.Subsequently,the mice were intraperitoneally injected with Klebsiella pneumoniae to establish bacterial infection.Serum and abdominal tissues were collected 48 h after infection.Hematoxylin-eosin(HE)staining was used to observe the pathological changes in the tissues,and enzyme-linked immunosorbent assay(ELISA)was used to measure the levels of inflammatory factors.In addition,the fecal samples collected before infection were analyzed for 16S rDNA gene of gut microbiota,and the expression levels of NF-κB/NLRP3 signaling pathway in colon tissues of uninfected mice were detected.[Results]Behavioral tests showed that compared with the control mice,CUMS mice had significantly lower body weight(P<0.0001,t=5.426),lower sucrose preference rate(P<0.001,t=4.937),increased swimming stationary time(P<0.001,t=16.37),and decreased time spent in the central area of the open field(P<0.01,t=3.575).Survival analysis showed that compared with the control mice,the survival rate of CUMS mice significantly decreased after infection(P<0.05).Additionally,histochemical staining showed that tissue damage in the liver(P<0.05,t=4.025),kidney(P<0.05,t=2.828),and mesentery(P<0.01,t=5.367)significantly increased.Furthermore,ELISA results showed that the levels of the inflammatory cytokines IL-6(P<0.01,t=3.365),IL-1β(P<0.01,t=4.061),TNF-α(P<0.01,t=4.460)and LPS(P<0.0001,t=27.24)were elevated.The difference was statistically significant.According to 16S rDNA sequencing,CUMS-induced changes in the intestinal bacterial community structure of mice,making them significantly different from the control mice.Compared with the control mice,the expression levels of NF-κB(P<0.01,t=6.825)and NLRP3(P<0.001,t=9.561)were upregulated in CUMS mice.[Conclusion]The CUMS model was successfully constructed and CUMS mice developed more severe bacterial infection.Gut microbiota was dysregulated and the expression of NF-κB/NLRP3 signaling pathway was up-regulated in CUMS mice,which was related to the susceptibility to bacterial infection.