OBJECTIVES: To study the impact of SURF4 expression level on long-term prognosis of gastric cancer (GC) and biological behaviors of GC cells. METHODS: SURF4 expression level in GC and its association with long-term patient prognosis were analyzed using publicly available databases and in 155 GC patients with low and high SURF4 expressions detected immunohistochemically. The Cox proportional hazard model and Kaplan-Meier survival curves were used to analyze independent prognostic predictors of GC and the 5-year survival rate of the patients with different SURF4 expression levels. Informatics analyses were conducted to explore the correlation of SURF4 expression level with immune cell infiltration in GC, SURF4-related differential genes and their associated pathways. In cultured GC cell line HGC-27, the effects of SURF4 knockdown and overexpression on proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) were investigated. RESULTS: Analysis of GEPIA dataset and immunohistochemical results suggested significant SURF4 overexpression in GC (P<0.05), which was associated with shortened 5-year survival time of the patients (χ²=38.749, P<0.001). The prognosis of GC was closely related to tumor stage T3-4, N2-3, CEA≥5 μg/L and CA19-9≥37 kU/L (P<0.05). SURF4 expression level was negatively correlated with activated B cells, NK cells and CD8⁺ effector memory T cells (P<0.05) and positively correlated with CD4⁺ T cells (P<0.05). GO and KEGG enrichment analysis suggested that SUFR4 may participate in GC carcinogenesis by promoting EMT through the tight junction pathway. In HGC-27 cells, SURF4 overexpression significantly decreased E-cadherin expression, increased N-cadherin expression, inhibited ZO-1 and claudin-1 expressions, and promoted cell proliferation, migration and invasion. CONCLUSIONS SURF4 is highly expressed in GC, and its overexpression is associated with a shortened 5-year survival of the patients possibly by enhancing tumor cell proliferation, migration and invasion via inhibiting tight junction proteins and promoting EMT.
Humans ; Stomach Neoplasms/metabolism* ; Cell Proliferation ; Cell Movement ; Epithelial-Mesenchymal Transition ; Cell Line, Tumor ; Neoplasm Invasiveness ; Prognosis ; Tight Junction Proteins/metabolism* ; Membrane Proteins/metabolism* ; Female ; Male

Objective To investigate the effect and mechanism of Xuanfu Daizhe Decoction on the proliferation,migration,and invasion activities of esophageal cancer cells.Methods Human e-sophageal cancer cell line EC 109 was treated with Xuanfu Daizhe Decoction at different concentrations,and the cells were divided into high-concentration group(200 μg/mL),medium-concentration group(100 μg/mL),low-concentration group(50 μg/mL),and blank group(0 μg/mL)based on the concentration.CCK-8 assay,wound healing assay,and Transwell invasion assay were used to detect the proliferation,migration,and invasion activities of the cells in each group,respectively.Western blot and cellular immunofluorescence staining were employed to detect the protein expression levels of glycolysis-related enzymes[hexokinase 2(HK2),lactate dehydrogenase A(LDHA),and phospho-fructokinase 1(PFK1)]in the cells of each group,and real-time quantitative fluorescent polymerase chain reaction(qRT-PCR)was used to detect the relative expression levels of mRNA encoding these enzymes.A nude mouse tumor-bearing model was established and divided into control group and Xuanfu Daizhe Decoction-fed group(20 mg/kg)to observe the effect of Xuanfu Daizhe Decoction on the growth of esophageal cancer in vivo.Results The results of CCK-8 assay,wound healing as-say,and Transwell invasion assay showed that Xuanfu Daizhe Decoction could inhibit the prolifera-tion,migration,and invasion activities of EC 109 cells in a concentration-dependent manner.West-ern blot and cellular immunofluorescence analysis revealed that compared with the blank group,the protein expression levels of HK2,LDHA,and PFK1 in the low-concentration,medium-concentra-tion,and high-concentration groups were decreased(P＜0.05).The qRT-PCR results showed that compared with the blank group,the relative expression levels of HK2 mRNA,LDHA mRNA,and PFK1 mRNA in the low-concentration,medium-concentration,and high-concentration groups were all reduced(P＜0.01 or P＜0.000 1).The tumor volume in the subcutaneous tissue on the back of nude mice in the Xuanfu Daizhe Decoction-fed group was smaller than that in the control group,and the protein expression level of LDHA in the tumor tissue of the Xuanfu Daizhe Decoction-fed group was lower than that in the control group,while the expression level of the pro-apoptotic protein Bax was higher than that in the control group(P＜0.05).Conclusion Xuanfu Daizhe Decoction can inhibit the glycolysis process of esophageal cancer cells in a concentration-dependent manner,there-by inhibiting cell proliferation,migration,invasion,and tumor growth in vivo.

BACKGROUND:Extracellular vesicles have received extensive attention in the field of bone defect regeneration and repair in recent years.However,natural extracellular vesicles have deficiencies in sustained controlled release,tissue targeting,and drug loading capacity.Therefore,the introduction of engineering strategies to modify extracellular vesicles to enhance their therapeutic efficacy has become a research hotspot.OBJECTIVE:To review the role and application progress of engineered extracellular vesicles in the regeneration and repair of bone defects.METHODS:PubMed,Web of Science,CNKI,and WanFang databases were searched for relevant articles published in the past fifteen years.The search terms were"engineering,extracellular vesicles,exosomes,bone defect,bone regeneration,bone repair"in Chinese and English.After removal of poorly related,outdated,and duplicate studies by screening,93 articles were finally included for review according to inclusion criteria.RESULTS AND CONCLUSION:(1)Extracellular vesicles are primarily isolated based on their density,size,immunoaffinity,and surface charge.After isolation,extracellular vesicles are characterized using imaging techniques,size-and counting-based techniques,and flow cytometry.(2)Extracellular vesicles stimulate bone regeneration by regulating immunity,angiogenesis,and proliferation and differentiation of target cells.(3)The engineering strategies of extracellular vesicles include surface modification and cargo loading.(4)The introduction of bone morphogenetic protein 2,mutant hypoxia-inducible factor-1α,vascular endothelial growth factor,miRNA and other bioactive factors into extracellular vesicles through engineering strategies can enhance their regenerative repair ability for bone defects.

BACKGROUND:Extracellular vesicles have received extensive attention in the field of bone defect regeneration and repair in recent years.However,natural extracellular vesicles have deficiencies in sustained controlled release,tissue targeting,and drug loading capacity.Therefore,the introduction of engineering strategies to modify extracellular vesicles to enhance their therapeutic efficacy has become a research hotspot.OBJECTIVE:To review the role and application progress of engineered extracellular vesicles in the regeneration and repair of bone defects.METHODS:PubMed,Web of Science,CNKI,and WanFang databases were searched for relevant articles published in the past fifteen years.The search terms were"engineering,extracellular vesicles,exosomes,bone defect,bone regeneration,bone repair"in Chinese and English.After removal of poorly related,outdated,and duplicate studies by screening,93 articles were finally included for review according to inclusion criteria.RESULTS AND CONCLUSION:(1)Extracellular vesicles are primarily isolated based on their density,size,immunoaffinity,and surface charge.After isolation,extracellular vesicles are characterized using imaging techniques,size-and counting-based techniques,and flow cytometry.(2)Extracellular vesicles stimulate bone regeneration by regulating immunity,angiogenesis,and proliferation and differentiation of target cells.(3)The engineering strategies of extracellular vesicles include surface modification and cargo loading.(4)The introduction of bone morphogenetic protein 2,mutant hypoxia-inducible factor-1α,vascular endothelial growth factor,miRNA and other bioactive factors into extracellular vesicles through engineering strategies can enhance their regenerative repair ability for bone defects.

Objective:To investigate the efficacy and safety of Doxycycline in the treatment of Mycoplasma pneumoniae pneumonia (MPP). Methods:Case series study.A retrospective observational study was conducted on 202 patients diagnosed with MPP in Beijing United Family Hospital from January 2022 to January 2024.The patients were divided into 3 groups according to antibiotics used: Azithromycin group which enrolled 93 cases, 60 cases (Azithromycin alone group) were included after excluding 33 cases with adding steroids; Doxycycline group which enrolled 32 case, 31 cases (Doxycycline alone group) were included after excluding 1 case with adding steroids; Azithromycin switch to Doxycycline when diagnosed Macrolides-unresponsive MPP(MUMPP). Azithromycin switch to Doxycycline group which enrolled 77 cases, 73 cases (switch group) were included after excluding 4 cases with adding steroids.Results used the Mann-Whitney U test, Chi-square test or exact test analyse, comparing the differences in defervescene time, hospitalization day, outcomes and side effects in each group. Results:The fever duration in the Doxycycline alone group was 48(36, 48) h, which was significantly shorter than that in the Azithromycin alone group[120(120, 144) h] ( Z=-7.646, P<0.001). The hospitalization time in the Doxycycline alone group was [3(3, 4) d], which was significantly shorter than that in the Azithromycin alone group [4(3, 5) d] ( Z=-3.368, P=0.002). The medium fever duration in the switch group after Azithromycin was changed to Doxycycline was 48 h, which was not statistically significantly different from that in the Doxycycline alone group ( Z=-0.571, P=0.849). The medium hospitalization time in the switch group after Azithromycin changed to Doxycycline was 4 d, which was significantly longer than that in the Doxycycline alone group (3 d) ( Z=-2.759, P=0.008). Among 93 cases enrolled in the Azithromycin group, 33 cases (35.5%) converted to unresponsive MPP(UMPP), 23 cases (24.7%) progressed to refractory MPP (RMPP), and 33 cases (35.5%) added steroids.Among 32 cases included in the Doxycycline group, 1 case (3.1%) converted to UMPP, no case developed to RMPP, and 1 case (3.1%) added steroids.The above-mentioned 3 proportions were significantly different between the two groups (all P<0.001). Among 93 cases enrolled in the Azithromycin group, 11 cases (11.8%) progressed to severe MPP (SMPP), and 13 cases (14.0%) developed complications.In 32 cases included in the Doxycycline group, 1 case (3.1%) progressed to SMPP, and 3 cases (9.4%) developed complications.The difference in these 2 proportions was not statistically significant between the two groups ( P=0.294, 0.760). In 77 patients included in the Azithromycin switch to Doxycycline group, there were 4 cases converting to UMPP, 4 cases converting to RMPP and 4 cases adding steroids, which were not statistically significantly different from those in the Doxycycline group (all P=0.540). In the Azithromycin switch to Doxycycline group, there were 3 cases progressing to SMPP, and 7 cases developed complications, which were not statistically significantly different from those in the Doxycycline group (all P=1.000). Conclusions:Doxycycline can improve the fever symptom, shorten illness duration and hospitalization time, and reduce steroid usage and the proportions of UMPP and RMPP in pediatric MPP.Switching to Doxycycline is recommended for MUMPP patients as Doxycycline is effective for the treatment of pediatric MPP and no tetracycline pigmentation teeth now.

Objective:To characterize the minimal inhibitory concentration (MIC) of macrolides against clinical Mycoplasma pneumoniae (MP) isolates and to investigate the significance of 23S rRNA mutations. Methods:Cross-sectional study.A total of 288 clinical MP strains preserved in the laboratory from 2016 to 2021 were taken for macrolide resistance gene testing and the evaluation of in vitro susceptibility to Azithromycin and Erythromycin.MIC 50 and MIC 90 values were calculated separately for macrolide-susceptible and -resistant strains. Results:All 288 MP strains underwent the test of in vitro susceptibility to Azithromycin, while 86 of them were additionally tested for Erythromycin.Among these strains, 22 strains were Azithromycin-sensitive, and 266 strains were Azithromycin-resistant.A2063G mutations were detected in 260 (97.7%) strains, while A2064G mutations were detected in 6 (2.3%) strains.Azithromycin-resistant strains had an MIC 50 of 128.000 μg/mL and an MIC 90 of 512.000 μg/mL, with the MIC ranging between 16.000 and 512.000 μg/mL.Seven strains were sensitive and 79 strains were resistant to Erythromycin.Among Erythromycin-resistant strains, A2063G mutations were detected in 73 (92.4%) strains, while A2064G mutations were detected in 6 (7.6%) strains.Erythromycin-resistant strains had an MIC 50 of 256.000 μg/mL and an MIC 90 of 512.000 μg/mL, with the MIC ranging between 64.000 and 1 024.000 μg/mL. Conclusions:A2063G and A2064G mutations in the 23S rRNA gene of MP are associated with high-level in vitro resistance to Azithromycin and Erythromycin, significantly limiting the clinical effectiveness of these antibiotics.Early resistance gene testing is recommended for suspected MP patients, which can help optimize the treatment, improve prognosis, and prevent resistance spread.

Objective:To investigate the efficacy and safety of Doxycycline in the treatment of Mycoplasma pneumoniae pneumonia (MPP). Methods:Case series study.A retrospective observational study was conducted on 202 patients diagnosed with MPP in Beijing United Family Hospital from January 2022 to January 2024.The patients were divided into 3 groups according to antibiotics used: Azithromycin group which enrolled 93 cases, 60 cases (Azithromycin alone group) were included after excluding 33 cases with adding steroids; Doxycycline group which enrolled 32 case, 31 cases (Doxycycline alone group) were included after excluding 1 case with adding steroids; Azithromycin switch to Doxycycline when diagnosed Macrolides-unresponsive MPP(MUMPP). Azithromycin switch to Doxycycline group which enrolled 77 cases, 73 cases (switch group) were included after excluding 4 cases with adding steroids.Results used the Mann-Whitney U test, Chi-square test or exact test analyse, comparing the differences in defervescene time, hospitalization day, outcomes and side effects in each group. Results:The fever duration in the Doxycycline alone group was 48(36, 48) h, which was significantly shorter than that in the Azithromycin alone group[120(120, 144) h] ( Z=-7.646, P<0.001). The hospitalization time in the Doxycycline alone group was [3(3, 4) d], which was significantly shorter than that in the Azithromycin alone group [4(3, 5) d] ( Z=-3.368, P=0.002). The medium fever duration in the switch group after Azithromycin was changed to Doxycycline was 48 h, which was not statistically significantly different from that in the Doxycycline alone group ( Z=-0.571, P=0.849). The medium hospitalization time in the switch group after Azithromycin changed to Doxycycline was 4 d, which was significantly longer than that in the Doxycycline alone group (3 d) ( Z=-2.759, P=0.008). Among 93 cases enrolled in the Azithromycin group, 33 cases (35.5%) converted to unresponsive MPP(UMPP), 23 cases (24.7%) progressed to refractory MPP (RMPP), and 33 cases (35.5%) added steroids.Among 32 cases included in the Doxycycline group, 1 case (3.1%) converted to UMPP, no case developed to RMPP, and 1 case (3.1%) added steroids.The above-mentioned 3 proportions were significantly different between the two groups (all P<0.001). Among 93 cases enrolled in the Azithromycin group, 11 cases (11.8%) progressed to severe MPP (SMPP), and 13 cases (14.0%) developed complications.In 32 cases included in the Doxycycline group, 1 case (3.1%) progressed to SMPP, and 3 cases (9.4%) developed complications.The difference in these 2 proportions was not statistically significant between the two groups ( P=0.294, 0.760). In 77 patients included in the Azithromycin switch to Doxycycline group, there were 4 cases converting to UMPP, 4 cases converting to RMPP and 4 cases adding steroids, which were not statistically significantly different from those in the Doxycycline group (all P=0.540). In the Azithromycin switch to Doxycycline group, there were 3 cases progressing to SMPP, and 7 cases developed complications, which were not statistically significantly different from those in the Doxycycline group (all P=1.000). Conclusions:Doxycycline can improve the fever symptom, shorten illness duration and hospitalization time, and reduce steroid usage and the proportions of UMPP and RMPP in pediatric MPP.Switching to Doxycycline is recommended for MUMPP patients as Doxycycline is effective for the treatment of pediatric MPP and no tetracycline pigmentation teeth now.

Objective:To characterize the minimal inhibitory concentration (MIC) of macrolides against clinical Mycoplasma pneumoniae (MP) isolates and to investigate the significance of 23S rRNA mutations. Methods:Cross-sectional study.A total of 288 clinical MP strains preserved in the laboratory from 2016 to 2021 were taken for macrolide resistance gene testing and the evaluation of in vitro susceptibility to Azithromycin and Erythromycin.MIC 50 and MIC 90 values were calculated separately for macrolide-susceptible and -resistant strains. Results:All 288 MP strains underwent the test of in vitro susceptibility to Azithromycin, while 86 of them were additionally tested for Erythromycin.Among these strains, 22 strains were Azithromycin-sensitive, and 266 strains were Azithromycin-resistant.A2063G mutations were detected in 260 (97.7%) strains, while A2064G mutations were detected in 6 (2.3%) strains.Azithromycin-resistant strains had an MIC 50 of 128.000 μg/mL and an MIC 90 of 512.000 μg/mL, with the MIC ranging between 16.000 and 512.000 μg/mL.Seven strains were sensitive and 79 strains were resistant to Erythromycin.Among Erythromycin-resistant strains, A2063G mutations were detected in 73 (92.4%) strains, while A2064G mutations were detected in 6 (7.6%) strains.Erythromycin-resistant strains had an MIC 50 of 256.000 μg/mL and an MIC 90 of 512.000 μg/mL, with the MIC ranging between 64.000 and 1 024.000 μg/mL. Conclusions:A2063G and A2064G mutations in the 23S rRNA gene of MP are associated with high-level in vitro resistance to Azithromycin and Erythromycin, significantly limiting the clinical effectiveness of these antibiotics.Early resistance gene testing is recommended for suspected MP patients, which can help optimize the treatment, improve prognosis, and prevent resistance spread.

Objective To investigate the dosimetric differences of different types of multileaf collima-tors(MLCs)on the planning target volume(PTV)and organs at risk(OARs)in volumetric modulated arc therapy(VMAT)for cervical cancer.Methods Twenty postoperative patients with cervical cancer receiving radiotherapy in this hospital from May 2023 to January 2024 were selected as the study subjects.For each pa-tient,two kinds of VMAT plans(MLCi2-MLC and AgilityTM-MLC)after cervical cancer operation were de-signed.The dosimetric parameters of the planning target volume(PTV),exposure dose by organs at risk(OAR),y passing rate and monitor units(MU)were compared between the two plans.Results Compared with MLci2-MLC,prescription dose(V45),conformity index(CI)and homogeneity index(HI)in AgilityTM-MLC were better,and the differences were statistically significant(P＜0.05),Compared with MLCi2-MLC,bladder V30,V40,average dose(Dmean),rectal V10,V20,V30,left femoral head V10,Dmean and small intestine V10,V30,Dmean and Dmax were lower,bladder V45,rectal V45,right femoral head V20 were higher,and the differences were statistically significant(P＜0.05).Compared with MLCi2-MLC,the y passing rate in AgilityTM-MLC was lower[(98.31±0.64)％vs.(99.73±0.37)％],and the difference was statistically significant(P＜0.05);while MU had no statistical difference between AgilityTM-MLC and MLCi2-MLC(996.74±65.46 vs.996.80±49.77,P＞0.05).Conclusion AgilityTM-MLC demonstrates better PTV conformity and homogenei-ty,as well as good protection on OARs in both high and low dose regions.

Purpose To investigate the expression of pro-tein of relevant evolutionary and lymphoid interest domain-con-taining 1(PRELID1)in gastric cancer tissues and to analyze its effect on prognosis,and the mechanism of influencing the prolif-eration and invasion ability of gastric cancer cells.Methods Using TCGA data and clinical data of 111 patients with gastric cancer,we analyzed the relationship between the expression of PRELID1 and clinicopathological parameters and the impact on clinical prognosis.The biological function of PRELID1 was pre-dicted by bioinformatics,and further verified by in vitro and in vivo experiments.Lentivirus was applied to regulate the level of PRELID 1 in gastric cancer cell line(MGC803)in vitro,and its effect on the proliferation,migration,and invasion of gastric cancer cells was observed.The nude mouse subcutaneous tumor-igenesis was used to observe the effect of PRELID1 on the growth of gastric cancer tissue in vivo.Results The expression of PRELID1 was significantly higher in gastric cancer tissues than that in the adjacent tissues(P＜0.001)and was positively cor-related with the cell proliferation indicator Ki67(P＜0.001).Cox regression model analysis showed that the high expression of PRELID 1 was an independent risk factor affecting the 5-year survival rate after radical gastrectomy(HR=2.336;95％CI=1.354-4.029).Gene enrichment results showed that the func-tion of PRELID1 was related to proliferation and JAK/STAT sig-naling.CCK-8 and Transwell experiments found that up-regula-tion of PRELID1 promoted the proliferation(P=0.016),mi-gration(P=0.016)and invasion(P=0.025)of gastric cancer cells,while down-regulation inhibited the proliferation(P=0.026),migration(P=0.048)and invasion(P=0.029).Subcutaneous tumor formation experiments in nude mice found that up-regulation of PRELID1 promoted the growth of gastric cancer tissue(P=0.047),while down-regulation was the oppo-site(P=0.005).Western blot detecting gastric cancer cells and gastric cancer tissues found that up-regulation of PRELID1 promoted the expression of JAK and STAT proteins(all P＜0.05),while down-regulation inhibited them(all P＜0.05).Conclusion The high expression of PRELID1 associated with poor prognosis may regulate the proliferation,migration and in-vasion of gastric cancer cells by up-regulating JAK/STAT signa-ling in gastric cancer.