OBJECTIVE: To investigate the effect of zedoarondiol on neovascularization of atherosclerotic (AS) plaque by exosomes experiment. METHODS: ApoE^-/- mice were fed with high-fat diet to establish AS model and treated with high- and low-dose (10, 5 mg/kg daily) of zedoarondiol, respectively. After 14 weeks, the expressions of anti-angiogenic protein thrombospondin 1 (THBS-1) and its receptor CD36 in plaques, as well as platelet activation rate and exosome-derived miR-let-7a were detected. Then, zedoarondiol was used to intervene in platelets in vitro, and miR-let-7a was detected in platelet-derived exosomes (Pexo). Finally, human umbilical vein endothelial cells (HUVECs) were transfected with miR-let-7a mimics and treated with Pexo to observe the effect of miR-let-7a in Pexo on tube formation. RESULTS: Animal experiments showed that after treating with zedoarondiol, the neovascularization density in plaques of AS mice was significantly reduced, THBS-1 and CD36 increased, the platelet activation rate was markedly reduced, and the miR-let-7a level in Pexo was reduced (P<0.01). In vitro experiments, the platelet activation rate and miR-let-7a levels in Pexo were significantly reduced after zedoarondiol's intervention. Cell experiments showed that after Pexo's intervention, the tube length increased, and the transfection of miR-let-7a minics further increased the tube length of cells, while reducing the expressions of THBS-1 and CD36. CONCLUSION Zedoarondiol has the effect of inhibiting neovascularization within plaque in AS mice, and its mechanism may be potentially related to inhibiting platelet activation and reducing the Pexo-derived miRNA-let-7a level.
Animals ; MicroRNAs/genetics* ; Exosomes/drug effects* ; Plaque, Atherosclerotic/genetics* ; Neovascularization, Pathologic/genetics* ; Human Umbilical Vein Endothelial Cells/metabolism* ; Humans ; Blood Platelets/drug effects* ; Apolipoproteins E/deficiency* ; Thrombospondin 1/metabolism* ; CD36 Antigens/metabolism* ; Platelet Activation/drug effects* ; Male ; Mice ; Mice, Inbred C57BL

Objective:This study aimed to provide basic data for the prevention, diagnosis and treatment of pediatric viral myocarditis (VMC) in China through analyzing the epidemic characteristics and disease burden of pediatric inpatients with VMC from 2016 to 2021.Methods:We performed a descriptive statistical analysis to the age, genders, seasons, regions and hospitalization cost and days of pediatric VMC inpatients and the death. All of the information was obtained from 27 Children′s hospitals or Maternal and Child Health hospitals of 23 provinces of China from 2016 to 2021.Results:A total of 7 647 599 cases including 1 646 VMC inpatients were admitted into our study. The annual numbers of hospitalizations were 173, 227, 313, 301, 295 and 337, with the hospitalized constituent ratios being 14.9/100 000, 17.9/100 000, 23.0/100 000, 20.5/100 000, 26.5/100 000 and 26.4/100 000 from 2016 to 2021. In recent 6 years, the proportion of VMC hospitalizations had increased yearly ( P<0.001), and had associated with the onset age ( P<0.001). Aged 12-≤18 years owned the highest hospitalized constituent ratio. The Northeast of China owned the largest number of VMC inpatients, and the East second to it. Among the 1 646 VMC children, there were 68 deaths, with the hospitalized case fatality rate of 4.13%. There were no significant differences between genders, age, seasons, years and fatality rate of VMC inpatients. For the diseases burden, the median of hospitalization days of all VMC inpatients was 10 days (IQR 6, 21), and the median of hospitalization cost was 1 1 842.3 RMB (IQR 6 969.22, 19 714.78). The median of hospitalization days of deceased VMC children was only 1 day (IQR 1, 3), the median cost could be 8 874.03 RMB (IQR 5 277.94, 5 6 151.59). Conclusions:In this study, we found that proportion of hospitalization of VMC children increased year by year, adolescence might be a risk factor of VMC. The fatality of VMC inpatients could be up to 4.13%, and the death led to a huge economic burden of society, family and individuals.

Objective To analyze the characteristics of males with autologous sperm preservation(ASP)in Anhui human sperm bank,and to explore the future direction of ASP in human sperm bank.Methods The basic infor-mation of males applied for ASP in Anhui human sperm bank from January 2019 to December 2023 was retrospec-tively analyzed.Results During this period,there were 424 males applied for ASP.93.40％(396/424)came from Anhui Province,of which 46.46％(197/424)came from Hefei.The age range of them was 15 to 59 years old.66.04％(280/424)had a college degree or above.23.11％(98/424)were employees of public institutions or enterprises.26.89％(114/424)were unmarried and 89.39％(379/424)were childless.67.45％(286/424)patients applied for ASP because of assisted reproductive technology treatment.15.33％(65/424)patients did it due to tumors,among which testicular cancer,lymphoma,leukemia and seminoma were the main reasons.A total of 1 163 semen samples were saved,and 53 males had used their sperm.Conclusion Only a few people applied for ASP,and the characteristics of males with ASP can be used to further strengthen publicity for key groups,espe-cially cancer patients,so as to benefit more people with autologous sperm preservation.

Objective:To observe the clinical effect of lower eyelid blepharoplasty combined with micro-autologous fat transplantation (MAFT) in the rejuvenation of the infraorbital regions.Method:A retrospective analysis was conducted on the clinical data of patients with lower eyelid pouch and tear trough treated with lower eyelid blepharoplasty combined with MAFT in the Department of Plastic Surgery of Affiliated Hospital of Nanjing University of Chinese Medicine from March 2020 to March 2022. Patients seeked medical attention due to lower eyelid pouch and tear trough, with or without skin laxity, and some patients had varying degrees of midface depression. After preoperative examination and evaluation of the patient’s infraorbital area and midfacial depression, appropriate individualized plans were developed.The patient underwent lower eyelid blepharoplasty by transconjunctival approach combined with tear trough MAFT. For patients with low or depressed midface, midface MAFT was also performed, and for patients with skin laxity, skin removal surgery was performed simultaneously. Complications were recorded after the operation. Preoperative and postoperative photos of more than 6 months were taken with the same postion and delivered to the same doctor to grade the patient’s Barton classification (0-3 grades, with higher grade indicating more obvious tear trough deformity). The improvement of tear trough deformity after surgery was assessed, and the global aesthetic improvement scale (GAIS) score was also assessed. The patients evaluated their improvement using the patients aesthetic improvement scale (PAIS) score (GAIS and PAIS are both 1-5 points, with higher scores indicating more significant postoperative improvement). Normal distribution metric data was represented by Mean±SD, and count data was represented by frequency and/or percentage; skewed distribution metric data was represented by M ( Q1, Q3), and compared by rank sum test. Results:A total of 69 patients were enrolled, including 9 males and 60 females, aged 25-45 years, with an average age of 36.7 years. Among them, 35 patients had midfacial depression, and 11 patients had skin laxity. (0.37±0.07) ml (0.2-0.6 ml) of micro-autologous fat was injected into the orbicularis oculi muscle on each side; (0.18±0.06) ml (0.1-0.2 ml) of micro-autologous fat was injected into the subcutaneous layer on each side; (3.80±0.98) ml (3-5 ml) of micro-autologous fat was injected into the deep medial cheek fat compartment and pyriform aperture fat compartment on each side. No serious complications occurred after the surgery. Postoperative follow-up lasted for 6-13 months, with an average of 8.7 months. The patient’s lower eyelid pouch, tear trough deformity, and depression in the midface have all improved, presenting a youthful and smooth appearance from the infraorbital area to the midface. The skin texture in the lower eyelid area had also improved. At the last follow-up, the Barton classification of the lacrimal groove decreased compared with that before surgery, and the difference was statistically significant [grade 0 (0, 1) vs. grade 2 (1, 2), Z=-7.62, P<0.001]. At the last follow-up, the PAIS score of 69 patients was 5 points in 49 cases (71%) and 4 points in 20 cases (29%); doctor’s GAIS score was 5 points in 38 cases (55%), 4 points in 27 cases (39%), and 3 points in 4 cases (6%) . Conclusion:For those patients with aging status of infraorbital region, like lower eyelid pouch and tear trough deformities, the combination of lower eyelid blepharoplasty through the transconjunctival approach and MAFT can achieve good therapeutic effects. This method is safe with a short recovery time.

Objective:To observe the clinical effect of lower eyelid blepharoplasty combined with micro-autologous fat transplantation (MAFT) in the rejuvenation of the infraorbital regions.Method:A retrospective analysis was conducted on the clinical data of patients with lower eyelid pouch and tear trough treated with lower eyelid blepharoplasty combined with MAFT in the Department of Plastic Surgery of Affiliated Hospital of Nanjing University of Chinese Medicine from March 2020 to March 2022. Patients seeked medical attention due to lower eyelid pouch and tear trough, with or without skin laxity, and some patients had varying degrees of midface depression. After preoperative examination and evaluation of the patient’s infraorbital area and midfacial depression, appropriate individualized plans were developed.The patient underwent lower eyelid blepharoplasty by transconjunctival approach combined with tear trough MAFT. For patients with low or depressed midface, midface MAFT was also performed, and for patients with skin laxity, skin removal surgery was performed simultaneously. Complications were recorded after the operation. Preoperative and postoperative photos of more than 6 months were taken with the same postion and delivered to the same doctor to grade the patient’s Barton classification (0-3 grades, with higher grade indicating more obvious tear trough deformity). The improvement of tear trough deformity after surgery was assessed, and the global aesthetic improvement scale (GAIS) score was also assessed. The patients evaluated their improvement using the patients aesthetic improvement scale (PAIS) score (GAIS and PAIS are both 1-5 points, with higher scores indicating more significant postoperative improvement). Normal distribution metric data was represented by Mean±SD, and count data was represented by frequency and/or percentage; skewed distribution metric data was represented by M ( Q1, Q3), and compared by rank sum test. Results:A total of 69 patients were enrolled, including 9 males and 60 females, aged 25-45 years, with an average age of 36.7 years. Among them, 35 patients had midfacial depression, and 11 patients had skin laxity. (0.37±0.07) ml (0.2-0.6 ml) of micro-autologous fat was injected into the orbicularis oculi muscle on each side; (0.18±0.06) ml (0.1-0.2 ml) of micro-autologous fat was injected into the subcutaneous layer on each side; (3.80±0.98) ml (3-5 ml) of micro-autologous fat was injected into the deep medial cheek fat compartment and pyriform aperture fat compartment on each side. No serious complications occurred after the surgery. Postoperative follow-up lasted for 6-13 months, with an average of 8.7 months. The patient’s lower eyelid pouch, tear trough deformity, and depression in the midface have all improved, presenting a youthful and smooth appearance from the infraorbital area to the midface. The skin texture in the lower eyelid area had also improved. At the last follow-up, the Barton classification of the lacrimal groove decreased compared with that before surgery, and the difference was statistically significant [grade 0 (0, 1) vs. grade 2 (1, 2), Z=-7.62, P<0.001]. At the last follow-up, the PAIS score of 69 patients was 5 points in 49 cases (71%) and 4 points in 20 cases (29%); doctor’s GAIS score was 5 points in 38 cases (55%), 4 points in 27 cases (39%), and 3 points in 4 cases (6%) . Conclusion:For those patients with aging status of infraorbital region, like lower eyelid pouch and tear trough deformities, the combination of lower eyelid blepharoplasty through the transconjunctival approach and MAFT can achieve good therapeutic effects. This method is safe with a short recovery time.

AIM:To investigate the effect of ankyrin repeat domain 49(ANKRD49)on epithelial-mesenchy-mal transition(EMT)in NCI-H1299 cells,and to explore its mechanism.METHODS:The ANKRD49 was over-ex-pressed in NCI-H1299 cells.The morphological changes of ANKRD49-overpressing NCI-H1299 cells were observed under microscope.The mRNA and protein expression levels of EMT-related markers[E-cadherin,transforming growth factor-β1(TGF-β1),vimentin and α-smooth muscle actin(α-SMA)]and EMT-related transcription factors(Snail1,Slug,Twist and ZEB1)were detected by RT-qPCR Western blot.Immunofluorescence staining was performed to observe the localiza-tion and expression of E-cadherin and vimentin in ANKRD49-overexpressing cells or control cells.Immunohistochemical method was performed to examine the levels of E-cadherin,α-SMA,Snail,Slug and ZEB1 in lung tissues of nude mice in-oculated with ANKRD49-overexpressing H1299 cells or control cells.RESULTS:Compared with the control group,the ANKRD49 overexpressing cells showed mesenchymal cell morphology(fusiform and less tight connections).RT-qPCR and Western blot results showed that the mRNA and protein levels of mesenchymal markers vimentin and α-SMA in ANKRD49 overexpressing cells were significantly higher than those in cells of control group,while the mRNA and protein levels of epithelial marker E-cadherin were lower than those in cells of control group.Compared with control group,the im-munofluorescence intensity of E-cadherin of H1299 cells decreased in after ANKRD49 overexpression,while that of vimen-tin increased significantly.Snail,Slug and ZEB1 expression were significantly elevated in ANKRD49 overexpressing cells compared with control group.The levels of E-cadherin in lung tissues of nude mice inoculated with ANKRD49-overexpressing H1299 cells declined,while the levels of α-SMA,Snail,Slug and ZEB1 increased compared with those in control mice.CONCLUSION:ANKRD49 promoted EMT of NCI-H1299 cells by increasing the expression of Snail1,Slug and ZEB1 and consequent downreguation of E-cadherin and upregulation of vimentin and α-SMA.

Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in tertiary hospitals in major regions of China in 2022.Methods Clinical isolates from 58 hospitals in China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2022 Clinical &Laboratory Standards Institute(CLSI)breakpoints.Results A total of 318 013 clinical isolates were collected from January 1,2022 to December 31,2022,of which 29.5％were gram-positive and 70.5％were gram-negative.The prevalence of methicillin-resistant strains in Staphylococcus aureus,Staphylococcus epidermidis and other coagulase-negative Staphylococcus species(excluding Staphylococcus pseudintermedius and Staphylococcus schleiferi)was 28.3％,76.7％and 77.9％,respectively.Overall,94.0％of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 90.8％of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis showed significantly lower resistance rates to most antimicrobial agents tested than Enterococcus faecium.A few vancomycin-resistant strains were identified in both E.faecalis and E.faecium.The prevalence of penicillin-susceptible Streptococcus pneumoniae was 94.2％in the isolates from children and 95.7％in the isolates from adults.The resistance rate to carbapenems was lower than 13.1％in most Enterobacterales species except for Klebsiella,21.7％-23.1％of which were resistant to carbapenems.Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.1％to 13.3％.The prevalence of meropenem-resistant strains decreased from 23.5％in 2019 to 18.0％in 2022 in Pseudomonas aeruginosa,and decreased from 79.0％in 2019 to 72.5％in 2022 in Acinetobacter baumannii.Conclusions The resistance of clinical isolates to the commonly used antimicrobial agents is still increasing in tertiary hospitals.However,the prevalence of important carbapenem-resistant organisms such as carbapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a downward trend in recent years.This finding suggests that the strategy of combining antimicrobial resistance surveillance with multidisciplinary concerted action works well in curbing the spread of resistant bacteria.

Objective To investigate the distribution and antimicrobial resistance profiles of the common pathogens isolated from urine from 2015 to 2021 in the CHINET Antimicrobial Resistance Surveillance Program.Methods The bacterial strains were isolated from urine and identified routinely in 51 hospitals across China in the CHINET Antimicrobial Resistance Surveillance Program from 2015 to 2021.Antimicrobial susceptibility was determined by Kirby-Bauer method,automatic microbiological analysis system and E-test according to the unified protocol.Results A total of 261 893 nonduplicate strains were isolated from urine specimen from 2015 to 2021,of which gram-positive bacteria accounted for 23.8％(62 219/261 893),and gram-negative bacteria 76.2％(199 674/261 893).The most common species were E.coli(46.7％),E.faecium(10.4％),K.pneumoniae(9.8％),E.faecalis(8.7％),P.mirabilis(3.5％),P.aeruginosa(3.4％),SS.agalactiae(2.6％),and E.cloacae(2.1％).The strains were more frequently isolated from inpatients versus outpatients and emergency patients,from females versus males,and from adults versus children.The prevalence of ESBLs-producing strains in E.coli,K.pneumoniae and P.mirabilis was 53.2％,52.8％and 37.0％,respectively.The prevalence of carbapenem-resistant strains in E.coli,K.pneumoniae,P.aeruginosa and A.baumannii was 1.7％,18.5％,16.4％,and 40.3％,respectively.Lower than 10％of the E.faecalis isolates were resistant to ampicillin,nitrofurantoin,linezolid,vancomycin,teicoplanin and fosfomycin.More than 90％of the E.faecium isolates were ressitant to ampicillin,levofloxacin and erythromycin.The percentage of strains resistant to vancomycin,linezolid or teicoplanin was＜2％.The E.coli,K.pneumoniae,P.aeruginosa and A.baumannii strains isolated from ICU inpatients showed significantly higher resistance rates than the corresponding strains isolated from outpatients and non-ICU inpatients.Conclusions E.coli,Enterococcus and K.pneumoniae are the most common pathogens in urinary tract infection.The bacterial species and antimicrobial resistance of urinary isolates vary with different populations.More attention should be paid to antimicrobial resistance surveillance and reduce the irrational use of antimicrobial agents.

Objective To understand the distribution and changing resistance profiles of clinical isolates of Enterococcus in hospitals across China from 2015 to 2021.Methods Antimicrobial susceptibility testing was conducted for the clinical isolates of Enterococcus according to the unified protocol of CHINET program by automated systems,Kirby-Bauer method,or E-test strip.The results were interpreted according to the Clinical & Laboratory Standards Institute(CLSI)breakpoints in 2021.WHONET 5.6 software was used for statistical analysis.Results A total of 124 565 strains of Enterococcus were isolated during the 7-year period,mainly including Enterococcus faecalis(50.7％)and Enterococcus faecalis(41.5％).The strains were mainly isolated from urinary tract specimens(46.9％±2.6％),and primarily from the patients in the department of internal medicine,surgery and ICU.E.faecium and E.faecalis strains showed low level resistance rate to vancomycin,teicoplanin and linezolid(≤3.6％).The prevalence of vancomycin-resistant E.faecalis and E.faecium was 0.1％and 1.3％,respectively.The prevalence of linezolid-resistant E.faecalis increased from 0.7％in 2015 to 3.4％in 2021,while the prevalence of linezolid-resistant E.faecium was 0.3％.Conclusions The clinical isolates of Enterococcus were still highly susceptible to vancomycin,teicoplanin,and linezolid,evidenced by a low resistance rate.However,the prevalence of linezolid-resistant E.faecalis was increasing during the 7-year period.It is necessary to strengthen antimicrobial resistance surveillance to effectively identify the emergence of antibiotic-resistant bacteria and curb the spread of resistant pathogens.