Objective To investigate the relationship between the levels of soluble FMS-like tyrosine ki-nase-1(sFlt-1),monocyte chemoatgulant protein-1(MCP-1),and soluble lectin-like oxidized low-density lipo-protein receptor 1(sLOX-1)in the serum of patients with coronary heart disease and the disease state and prognosis.Methods A total of 126 patients with coronary heart disease treated in Zhongshan Hospital,Dalian University from May 2022 to May 2024 were selected as study objects.The patients were divided into good prognosis group(n=78)and poor prognosis group(n=48)according to prognostic results.The serum sFlt-1,MCP-1 and sLOX-1 levels were detected by enzyme-linked immunosorbent assay.The degree of coronary artery stenosis in patients was measured by coronary angiography,as indicated by the Gensini score.Spearman correlation analysis was used to analyze the correlation between Gensini score and serum sFlt-1,MCP-1 and sLOX-1 levels.Multivariate Logistic regression was used to analyze the factors affecting the prognosis of pa-tients with coronary heart disease.The predictive efficacy of serum sFlt-1,MCP-1 and sLOX-1 levels for the prognosis of patients with coronary heart disease was evaluated by receiver operating characteristic(ROC)curve.Results The age in the poor prognosis group was older than that in the good prognosis group(P＜0.05),Gensini score was higher than that in the good prognosis group(P＜0.05).Compared with the poor prognosis group,the serum sFlt-1,MCP-1 and sLOX-1 levels in the good prognosis group were lower(P＜0.05).Gensini score was positively correlated with the serum sFlt-1,MCP-1 and sLOX-1 levels.Multivariate Logistic regression analysis showed that old age,high Gensini score,high sFlt-1 level,high MCP-1 level and high sLOX-1 level were all risk factors effecting the prognosis of patients with coronary heart disease(P＜0.05).ROC curve analysis showed that the areas under the curve(AUC)of serum sFlt-1,MCP-1,and sLOX-1 for predicting the prognosis of coronary heart disease patients were 0.787(95％CI:0.703-0.870),0.815(95％CI:0.741-0.890)and 0.795(95％CI:0.715-0.876),respectively.The AUC for predicting the prognosis of coronary heart disease patients using a combination of three was 0.923(95％CI:0.876-0.970).Conclusion The serum levels of sFlt-1,MCP-1 and sLOX-1 in patients with coronary heart disease are signifi-cantly increased,and are positively correlated with the degree of coronary artery stenosis.The levels of sFlt-1,MCP-1 and sLOX-1 in serum have a certain predictive effect on the prognosis of patients with coronary heart disease.

Objective:To explore the role of circ_0007766 molecule cyclized from the erb-b2 receptor tyrosine kinase 2(ERBB2)gene in prostate cancer(PCa)cells and the impact of hypoxia on the regulation of its expression.Methods:The expression of circ_0007766 molecule in PCa cells and tissues was detected by qRT-PCR.The siRNA of circ0007766 was transfected to PCa cells(DU145 cell and PC3 cell)respectively;Colony formation assay,CCK-8 assay and Transwell assay were performed to detect the effect of circ_0007766 knockdown on the colony formation,proliferation,migration,and invasion abilities of PCa cells.Hypoxia cell model of DU145 and PC3 cells were established by hypoxic chamber method,and WB was performed to detect the protein expression of HIF-1α,a key molecule of the hypoxic pathway.qRT-PCR was performed to detect the expression of circ_0007766 molecule in hypoxic cell model,and the protein expression of RNA-binding protein eukaryotic translation initiation factor 4A3(EIF4A3)was detected by WB.RNA immunoprecipitation(RIP)was performed to detect the binding of EIF4A3 to circ_0007766 under hypoxic conditions.qRT-PCR assay was performed to further detect the effect of EIF4A3 knockdown on the expression of circ_0007766 under hypoxic conditions.Results:circ_0007766 was highly expressed in PCa cells(P<0.01)and PCa tissues(P<0.05).The knockdown of circ_0007766(P<0.05 or P<0.01)could significantly inhibit the proliferation,migration and invasion abilities of PCa cells(all P<0.01).The upregulation of HIF-1α protein under hypoxic conditions confirmed the successful establishment of the hypoxia cell model.qRT-PCR analysis revealed that compared with that in the normoxia group,circ_0007766 expression was markedly elevated in the hypoxia group(P<0.01).WB analysis demonstrated increased EIF4A3 protein expression in cells of the hypoxia group.RIP assays indicated that circ_0007766 was highly concentrated in the EIF4A3-enriched group(P<0.01).Additionally,qRT-PCR showed that hypoxia significantly boosted circ_0007766 expression,whereas EIF4A3 knockdown notably diminished the hypoxia-induced expression of circ_0007766(P<0.05 or P<0.01).Conclusion:Circ-0007766 plays the role of cancer-promoting molecule in PCa cells,and its expression is related to the regulation of EIF4A3 molecule under hypoxic conditions.

Objective:To assess the effect of Hood's technique on urinary continence in patients undergoing single-port robot-assisted radical prostatectomy (spRARP).Methods:The clinical data of 53 patients who underwent spRARP performed by a single surgeon in the First Affiliated Hospital of Harbin Medical University from June 2021 to October 2023 were retrospectively analyzed. There were 25 patients in the spRARP+ Hood group and 28 patients in the spRARP group. There were no statistically significant differences between the spRARP+ Hood group and the spRARP group in terms of patients′ age [(70.28±5.98) years vs. (60.89±6.86) years old], body mass index[(24.64±2.85) kg/m 2 vs. (24.59±3.17) kg/m 2], prostate weight [70.00 (40.69, 102.25) g vs. 73.50 (49.13, 94.50) g], total prostate specific antigen[8.62 (4.56, 15.26) ng/ml vs. 12.68 (6.99, 19.24) ng/ml], Gleason score [8 (7, 8) vs. 8 (7, 8)], age-adjusted Charlson comorbidity index (aCCI) [4 (3, 5) vs. 3 (3, 4)], and clinical stage [T 2a/T 2b/T 2c: 6/10/9 cases vs. 5/7/16 cases ] ( P>0.05). In the SpRARP + Hood group, the detrusor apron, tendon arch, pubic prostatic ligament, and dorsal vascular complex were completely preserved during the operation. In contrast, this was not the case for spRARP.Additionally, the incision size, bleeding volume, intraoperative blood transfusion volume, operation time, gastrointestinal function recovery time, total amount of drainage in the first three postoperative days, retention time of the drainage tube, postoperative hospitalization time, positive incision margins, recovery rate of urinary continence immediately after removal of the urinary catheter, and the recovery rate of urinary continence at 1, 3, and 6 months postoperatively were compared between the two groups. Results:All 53 surgeries were successfully completed. The differences in incision length [4.0 (3.5, 4.0) cm vs. 4.0 (4.0, 4.0) cm], intraoperative bleeding [50 (40, 100) ml vs. 100 (50, 100) ml], and intraoperative transfusion rate [4.0% (1/25) vs. 17.8% (5/28)] were not statistically significant between the spRARP+ Hood group and the spRARP group ( P>0.05), and the difference in operative time [205.0 (167.5, 240.0) min vs. 242.5 (185.0, 300.0) min] was statistically significant( P<0.05).The recovery time of gastrointestinal function in the spRARP+ Hood group vs. the spRARP group [1.0 (1.0, 1.5) d vs. 1.0 (1.0, 2.0) d], total amount of drainage in the first 3 d postoperatively [150.00 (72.50, 261.00) ml vs. 230.00 (115.00, 417.50) ml], duration of drain retention [5.0 ( 4.0, 5.0) d vs. 5.0 (4.0, 6.8) d], postoperative hospital stay [5.0 (4.0, 7.5) d vs. 5.0 (3.3, 7.8) d], and margin positivity rate [4.0% (1/25) vs. 3.6% (1/28)] were not statistically significant ( P>0.05). The postoperative Clavien-Dindo complication classification was grade I in both groups. The differences between the spRARP+ Hood group and the spRARP group in the rates of recovery of urinary continence immediately after the urinary catheter removal [56.0% (14/25) vs. 7.1% (2/28)] and one month after surgery [76.0% (19/25) vs. 28.5% (8/28)] were statistically significant ( P<0.05). The differences in the rates of recovery of urinary continence at 3 months after surgery [80.0% (20/25) vs. 67.8% (19/28)], at 6 months after surgery [88.0% (22/25) vs. 96.4% (27/28)], and biochemical recurrence at 6 months after surgery [4.0% (1/25) vs. 3.6% (1/28)] were not statistically significant ( P>0.05). Conclusions:The outcomes of spRARP+ Hood in the treatment of localized prostate cancer were comparable to those of spRARP. However, spRARP+ Hood has better urinary continence immediately after removal of the urinary catheter and 1 month postoperatively.

Objective:To assess the effect of Hood's technique on urinary continence in patients undergoing single-port robot-assisted radical prostatectomy (spRARP).Methods:The clinical data of 53 patients who underwent spRARP performed by a single surgeon in the First Affiliated Hospital of Harbin Medical University from June 2021 to October 2023 were retrospectively analyzed. There were 25 patients in the spRARP+ Hood group and 28 patients in the spRARP group. There were no statistically significant differences between the spRARP+ Hood group and the spRARP group in terms of patients′ age [(70.28±5.98) years vs. (60.89±6.86) years old], body mass index[(24.64±2.85) kg/m 2 vs. (24.59±3.17) kg/m 2], prostate weight [70.00 (40.69, 102.25) g vs. 73.50 (49.13, 94.50) g], total prostate specific antigen[8.62 (4.56, 15.26) ng/ml vs. 12.68 (6.99, 19.24) ng/ml], Gleason score [8 (7, 8) vs. 8 (7, 8)], age-adjusted Charlson comorbidity index (aCCI) [4 (3, 5) vs. 3 (3, 4)], and clinical stage [T 2a/T 2b/T 2c: 6/10/9 cases vs. 5/7/16 cases ] ( P>0.05). In the SpRARP + Hood group, the detrusor apron, tendon arch, pubic prostatic ligament, and dorsal vascular complex were completely preserved during the operation. In contrast, this was not the case for spRARP.Additionally, the incision size, bleeding volume, intraoperative blood transfusion volume, operation time, gastrointestinal function recovery time, total amount of drainage in the first three postoperative days, retention time of the drainage tube, postoperative hospitalization time, positive incision margins, recovery rate of urinary continence immediately after removal of the urinary catheter, and the recovery rate of urinary continence at 1, 3, and 6 months postoperatively were compared between the two groups. Results:All 53 surgeries were successfully completed. The differences in incision length [4.0 (3.5, 4.0) cm vs. 4.0 (4.0, 4.0) cm], intraoperative bleeding [50 (40, 100) ml vs. 100 (50, 100) ml], and intraoperative transfusion rate [4.0% (1/25) vs. 17.8% (5/28)] were not statistically significant between the spRARP+ Hood group and the spRARP group ( P>0.05), and the difference in operative time [205.0 (167.5, 240.0) min vs. 242.5 (185.0, 300.0) min] was statistically significant( P<0.05).The recovery time of gastrointestinal function in the spRARP+ Hood group vs. the spRARP group [1.0 (1.0, 1.5) d vs. 1.0 (1.0, 2.0) d], total amount of drainage in the first 3 d postoperatively [150.00 (72.50, 261.00) ml vs. 230.00 (115.00, 417.50) ml], duration of drain retention [5.0 ( 4.0, 5.0) d vs. 5.0 (4.0, 6.8) d], postoperative hospital stay [5.0 (4.0, 7.5) d vs. 5.0 (3.3, 7.8) d], and margin positivity rate [4.0% (1/25) vs. 3.6% (1/28)] were not statistically significant ( P>0.05). The postoperative Clavien-Dindo complication classification was grade I in both groups. The differences between the spRARP+ Hood group and the spRARP group in the rates of recovery of urinary continence immediately after the urinary catheter removal [56.0% (14/25) vs. 7.1% (2/28)] and one month after surgery [76.0% (19/25) vs. 28.5% (8/28)] were statistically significant ( P<0.05). The differences in the rates of recovery of urinary continence at 3 months after surgery [80.0% (20/25) vs. 67.8% (19/28)], at 6 months after surgery [88.0% (22/25) vs. 96.4% (27/28)], and biochemical recurrence at 6 months after surgery [4.0% (1/25) vs. 3.6% (1/28)] were not statistically significant ( P>0.05). Conclusions:The outcomes of spRARP+ Hood in the treatment of localized prostate cancer were comparable to those of spRARP. However, spRARP+ Hood has better urinary continence immediately after removal of the urinary catheter and 1 month postoperatively.

Objective:To investigate differences in bacterial and fungal microbiome between infected nails and healthy nails among patients with onychomycosis.Methods:Nail scraping samples were collected from infected nails and healthy nails of 31 patients with onychomycosis, who visited Dalian Dermatosis Hospital from August 2020 to July 2021. The total DNA of nail microbiota was extracted, and the V3-V4 regions of the bacterial 16S rDNA gene and the fungal internal transcribed spacer (ITS) region were amplified and sequenced using Illumina technology. The USEARCH and mothur softwares were used for data cluster analysis to obtain the operational taxonomic units (OTUs) , Wilcoxon rank sum test was used to analyze α diversity, analysis of similarities (ANOSIM) was performed to analyze β diversity, linear discriminant analysis of effect size (LEfSe) was performed to evaluate the species difference.Results:Among the 31 patients with onychomycosis, 16 were males and 15 were females. According to the age, they were divided into young group (18 - 35 years old, 10 cases) , middle-aged group (36 - 60 years old, 11 cases) , and elderly group (over 60 years old, 10 cases) . As the α-diversity analysis revealed, the infected nail group showed significantly decreased Shannon index ( W = 290, P = 0.007) , but significantly increased Simpson index ( W = 663, P = 0.010) compared with the healthy nail group, suggesting that the diversity and evenness of bacterial communities were lower in the infected nail group than in the healthy nail group; however, there was no significant difference in the diversity of fungal communities between the infected nail group and healthy nail group. The β-diversity analysis based on the unweighted-UniFrac distance matrix showed no significant difference in the fungal or bacterial community composition between the infected nail group and healthy nail group (bacterial communities: R = 0.0052, P = 0.331; fungal communities: R = 0.0036, P = 0.337) ; the β-diversity analysis based on the weighted-UniFrac distance matrix showed significant differences in the abundance of bacterial and fungal communities between the two groups (both P = 0.001) . In terms of the species composition, the bacterial flora with significantly decreased abundance in the infected nail group compared with the healthy nail group included Bacteroidetes, Proteobacteria, Betaproteobacteria, Burkholderiales, Ralstonia, Sphingomonas and Streptococcus, while the abundance of Bacilli, Bacillales and Staphylococcus was significantly higher in the infected nail group than in the healthy nail group. Compared with the healthy nail group, the fungal flora with significantly increased abundance in the infected nail group included Eurotiomycetes, Onygenales, Leotiomycetes-ord-incertae-sedis, Arthrodermataceae, Periconia, Erysiphe, Tilletiopsis, Trichophyton, Erysiphe cruciferarum, Trichophyton rubrum, Malassezia sympodialis, while the abundance of Saccharomycetes, Saccharomycetales, Saccharomycetaceae, Dothioraceae, Candida and Alternaria was significantly lower in the infected nail group than in the healthy nail group. Conclusion:The diversity and abundance of bacterial communities significantly differed between infected nails and healthy nails among patients with onychomycosis, while only the abundance of fungal communities differed between the two groups, and perhaps there was correlations between some bacterial and fungal communities.

Objective To evaluate the feasibitity of robot-assisted posterior laparoscopic modified"single-position"radical nephroureterectomy.Methods A retrospective analysis was made on 7 patients receiving robot-assisted posterior laparoscopic single-position radical nephroureterectomy between April 2022 and April 2023.The patients were in a fully healthy lateral position,and an artificial pneumoperitoneum was established.Trocars were placed at the right costal margin of the posterior axillary line,3-4 cm above the iliac crest of the midaxillary line,6-8 cm below the anterior axillary line,and 3-4 cm above the iliac crest of the midaxillary line near the outer edge of the musculus rectus abdominis,respectively.After the kidney was removed,the ureter was freed down to the iliac vessels,and then the main joint of the robot was reversed 180° for redocking.The ureter was continuously freed downwards to the bladder wall and the catheter was clamped.The bladder was opened after filling with indocyanine green and distilled water mixture.Then the fluid in the bladder was washed,the contralateral ureteral orifice was observed,the affected side of the ureter was resected,and the bladder incision was sutured by two layers with V-LOCK 2-0 sutures.The incision was extended under the right costal margin of the posterior axillary line and 3-4 cm above the iliac crest of the midaxillary line to remove the specimen.Results The operation was successfully completed in all the 7 cases.The surgical operation time was 155-263 min(mean,247.0 min)and the blood loss was 20-100 ml(mean,42.9 ml).The postoperative anal exhaust time was 14-24 h(mean,22.6 h).There were 1 case of postoperative absorption fever,2 cases of moderate anemia,and 2 cases of postoperative incision fat liquefaction.In the 2 patients with moderate anemia,one patient developed postoperative intramuscular artery rupture leading to massive bleeding and the formation of hematoma in the surgical area,with the amount of bleeding being approximately 1000 ml,and the other had moderate anemia before and after surgery.The hospital stay ranged 8-16 d(mean,11.6 d).Pathologic examinations showed high-grade uroepithelial carcer in all the patients.Postoperative follow-ups lasted 3-9 months,with a mean of 6.2 months.None had bladder tumor recurrence or distant metastasis.Conclusion Robot-assisted posterior laparoscopic modified"single-position"radical nephroureterectomy is safe and feasible.

In recent years, the genome editing technologies based on the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein (CRISPR/Cas) have developed rapidly. The system can use homologous directed recombination (HDR) to achieve precise editing that it medicated, but the efficiency is extremely low, which limits its application in agriculture and biomedical fields. As an emerging genome editing technology, the CRISPR/Cas-mediated DNA base editing technologies can achieve targeted mutations of bases without generating double-strand breaks, and has higher editing efficiency and specificity compared with CRISPR/Cas-mediated HDR editing. At present, cytidine base editors (CBEs) that can mutate C to T, adenine base editors (ABEs) that can mutate A to G, and prime editors (PEs) that enable arbitrary base conversion and precise insertion and deletion of small fragments, have been developed. In addition, glycosylase base editors (GBEs) capable of transitioning from C to G and double base editors capable of editing both A and C simultaneously, have been developed. This review summarizes the development, advances, advantages and limitations of several DNA base editors. The successful applications of DNA base editing technology in biomedicine and agriculture, together with the prospects for further optimization and selection of DNA base editors, are discussed.
Agriculture ; CRISPR-Cas Systems/genetics* ; DNA/genetics* ; Gene Editing ; Technology

Objective:To investigate the effects of body mass index (BMI) levels at different baseline on the risk of new-onset acute pancreatitis (AP).Methods:The subjects were from the Kailuan Study Cohort and divided into 3 groups according to baseline BMI levels: BMI<24 kg/m 2, normal weight; BMI 24-28 kg/m 2, overweight; BMI≥28 kg/m 2, obesity. The incidence of new-onset AP in these three groups was analyzed. The survival curve was plotted by Kaplan-Meier method, the cumulative incidence was calculated and tested by log-rank method. Multivariate Cox proportional hazards regression model was used to calculate HR of baseline BMI levels for AP. Results:A total of 123 841 subjects were included and followed up for (11.94±2.13) years, during which, 395 cases were found with AP. The incidence of AP was 2.67 per 10 000 person years in total population, and the incidences of AP were 2.20, 2.72 and 3.58 per 10 000 person-years in the normal, overweight and obesity groups, respectively. The cumulative incidences of AP was 0.32%, 0.40% and 0.49% in normal, overweight and obesity groups, respectively, which showed a significant inter-group difference by log-rank test ( χ2=13.17, P<0.01). The results of multivariable adjusted Cox proportional hazards regression model analysis indicated that obesity group ( HR=1.45, 95% CI: 1.10-1.92) had a higher risk for AP compared with the normal BMI group. The subgroup analyses by age and sex showed that compared with the normal weight group,the HRs for AP in the obesity group was 1.58(95% CI:1.14-2.19) and 1.40(95% CI:1.03-1.90) among subjects younger than 60 years old and male subjects, respectively. After excluded onset AP within two years from baseline,with a control group from normal weight,the results of multivariate Cox proportional hazards regression model analysis indicated that the AP in the obesity group was 1.60 (95% CI: 1.18-2.15). Conclusion:Obesity may increase the risk of developing AP, particularly among young and middle-aged men.

Objective:To analyze the subcellular localization of family of serine hydrolases 1 (FSH1) protein in Microsporum canis. Methods:The FSH1 and enhanced green fluorescent protein (EGFP) genes were amplified by PCR using the previously constructed plasmid containing the FSH1 gene and the recombinant plasmid pCAMBIA-LRP-EGFP as the template; the vector DNA was obtained by double-enzyme digestion of the recombinant plasmid pCAMBIA-LRP-EGFP with SnaBI/KpnI. Then, the EGFP expression plasmid and Ptrcp-FSH1-EGHP-Ttrcp fusion plasmid were constructed by inserting the amplified EGFP gene and EGFP-FSH1 gene into the vector DNA respectively, and identified by PCR and sequencing. The two recombinant plasmids were transformed into Microsporum canis by an Agrobacterium tumefaciens-mediated method, and the gene EGFP and fusion gene FSH1-EGFP were expressed integratedly in Microsporum canis under the regulation by the fungal universal promoter Ptrpc and terminator Ttrpc. The cellular localization of the fusion protein was observed by laser scanning confocal microscopy. Results:The Agrobacterium tumefaciens-mediated transformation system and EGFP expression vector in Microsporum canis were successfully constructed; the fusion gene FSH1-EGFP was expressed integratedly in Microsporum canis. Laser confocal microscopy showed that fluorescence signals of the FSH1-EGFP fusion protein were concentrated in the cytoplasm and nuclei of Microsporum canis, with a granular or cluster-like appearance. Conclusion:The FSH1-EGFP fusion protein was successfully localized in the cytoplasm and nuclei of Microsporum canis, providing a basis for further clarifying the function and pathogenic mechanisms of the FSH1 gene in Microsporum canis.

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated proteins (Cas) system is a hotspot of gene editing and gene expression research, in which CRISPR/Cas13 system provides a new direction for RNA interference and editing. In this study, we designed and synthesized the corresponding gRNAs of CRISPR/Cas13a and CRISPR/Cas13b systems in non-homologous end joining (NHEJ) pathway, such as Ku70 and Lig4, and then detected the expression of ku70 and lig4 in HEK293T cells. The CRISPR/Cas13a system could efficiently knockdown the mRNA expression of ku70 and lig4 more than 50%, and CRISPR/Cas13b system also suppressed ku70 and lig4 about 92% and 76%, respectively. Also, CRISPR/Cas13a, b systems could down-regulate Ku70 and Lig4 proteins level to 68% and 53%, respectively. The study demonstrates that the CRISPR/Cas13 system could effectively knockdown the expression of RNA and protein in HEK293T cells, providing a new strategy for gene function and regulation research.
CRISPR-Cas Systems ; DNA Ligase ATP ; genetics ; Gene Expression Regulation ; genetics ; Gene Knockdown Techniques ; HEK293 Cells ; Humans ; Ku Autoantigen ; genetics