AIM: To develop a novel gene-delivery therapeutic based on CRISPR/Cas9 genome editing technology capable of specifically targeting and knocking out the VEGFA gene, thereby achieving sustained suppression of VEGFA expression in retinal pigment epithelial(RPE)cells and providing a new strategy for gene therapy in retinal neovascular diseases.METHODS:Single guide RNAs targeting the human VEGFA gene for knockout were designed, and corresponding recombinant plasmids were constructed. A novel polymer(PTEE)was used to encapsulate the plasmids to prepare a PTEE-loaded anti-VEGFA plasmid(PLAP)gene delivery system. PTEE materials at concentrations of 0.1, 0.2, 0.4, 0.8, and 1.6 μg/μL were co-incubated with ARPE-19 cells, and the biocompatibility of PTEE was evaluated using the cell counting kit-8(CCK-8)assay. Recombinant plasmids expressing green fluorescent protein(GFP)were constructed. Lipofectamine 3000 and jetOPTIMUS®DNA transfection reagents were used as control groups, and PTEE nanomaterials were used as the experimental group to encapsulate the plasmids. When the cell confluence reached 80%, the formulations were transfected into ARPE-19 and 293T cells. GFP expression was observed under light microscopy, and the transfection efficiencies of each group were compared. ARPE-19 cells were induced under hypoxia, and PLAP was transfected into the cells. The expression level of VEGFA was detected by enzyme-linked immunosorbent assay(ELISA)to evaluate the efficacy of this novel gene delivery system.RESULTS: After co-incubation of ARPE-19 cells with different concentrations of PTEE for 24 h and 48 h, no significant effect on cell viability was observed in any group. The transfection efficiency of PLAP in ARPE-19 cells was higher than that in the Lipo3000 and jetOPTIMUS groups, with statistically significant differences(P<0.01). Hypoxia for 6 h significantly induced the upregulation of VEGFA mRNA expression in ARPE-19 cells, and under hypoxic conditions, the PTEE group exhibited a significant inhibitory effect on VEGFA expression(P<0.01).CONCLUSION:PLAP exhibits favorable biocompatibility and prominent VEGFA inhibitory effects in vitro, making it a potential candidate drug for gene therapy of retinal neovascular diseases.

Objective To introduce an innovative technique, the "balance-shaped sternal elevation device" and its application in the subxiphoid uniportal video-assisted thoracoscopic surgery (VATS) for anterior mediastinal masses resection. Methods Patients who underwent single-port thoracoscopic assisted anterior mediastinal tumor resection through the xiphoid process at the Department of Thoracic Surgery, West China Hospital, Sichuan University from May to June 2024 were included, and their clinical data were analyzed. Results A total of 7 patients were included, with 3 males and 4 females, aged 28-72 years. The diameter of the tumor was 1.9-17.0 cm. The operation time was 62-308 min, intraoperative blood loss was 5-100 mL, postoperative chest drainage tube retention time was 0-9 days, pain score on the 7th day after surgery was 0-2 points, and postoperative hospital stay was 3-12 days. All patients underwent successful and complete resection of the masses and thymus, with favorable postoperative recovery. Conclusion The "balance-shaped sternal elevation device" effectively expands the retrosternal space, providing surgeons with satisfactory surgical views and operating space. This technique significantly enhances the efficacy and safety of minimally invasive surgery for anterior mediastinal masses, reduces trauma and postoperative pain, and accelerates patient recovery, demonstrating important clinical significance and application value.

AIM: To compare the changes in corneal epithelial thickness(CET)after small incision lenticule extraction(SMILE)and femtosecond laser-assisted in situ keratomileusis(FS-LASIK).METHODS: A total of 187 patients(187 eyes)who underwent either SMILE or FS-LASIK at Urumqi Aier Eye Hospital between December 2022 and November 2023 were collected. The patients were divided into SMILE group and FS-LASIK group according to surgical methods. The CET of the patients was measured by optical coherence tomography(OCT)system before surgery and at 1 wk, 1, 3, and 6 mo postoperatively.RESULTS: Changes in corneal epithelial thickness(△CET)in the central, paracentral, and mid-peripheral regions were compared at 6 mo postoperatively. The SMILE group was characterized by the most significant thickening in the central area and the least thickening in the mid-peripheral area; while the FS-LASIK group was characterized by the most significant thickening in the paracentral area and the least thickening in the mid-peripheral region. At 1 wk, 1, 3, and 6 mo postoperatively, within the 0-7 mm corneal area, the △CET for both the SMILE and FS-LASIK groups was correlated with the preoperative spherical equivalent.CONCLUSION: Within 6 mo postoperatively, both SMILE and FS-LASIK showed a similar trend in epithelial thickening but with distinct characteristics. The change in corneal epithelial thickness for both procedures was positively correlated with the preoperative diopter.

Objective:To explore the epidemiological characteristics, clinical manifestations, laboratory findings, and imaging features of children with HRV-associated pneumonia, and to analyze the clinical features and risk factors associated with severe HRV pneumonia, providing references for clinical management.Methods:A single-center, retrospective, observational study was conducted, including 1 001 cases of HRV-positive children with pneumonia admitted to the Respiratory Department of the Affiliated Children′s Hospital of Capital Institute of Pediatrics from January 2019 to December 2023. Among them, 584 cases (58.3%) were male and 417 cases (41.7%) were female, with an age range of 0.1 to 14.9 years, a median age of 3.42 years, and a mean age of (3.92±2.75) years. According to clinical guidelines, the cases were divided into a mild pneumonia group (855 cases, 510 males, 345 females) and a severe pneumonia group (146 cases, 73 males, 73 females). Basic information, clinical, laboratory, and imaging data were collected from the electronic medical record system. Comparisons between different age groups, diagnoses, and pneumonia severity groups were performed using the χ2 test. Multivariate logistic regression analysis was used to identify risk factors for the severity of HRV pneumonia. Results:Among the 1 001 cases of HRV-associated bronchopneumonia, 146 cases (14.6%) were severe pneumonia. The age of severe HRV pneumonia patients was significantly higher than that of the mild pneumonia group (5.2 years vs. 3.7 years, t=-6.050, P<0.01). Severe HRV pneumonia had a higher incidence in autumn and winter (60.9%). Severe HRV pneumonia was associated with higher levels of lactate dehydrogenase (LDH), C-reactive protein (CRP), neutrophils, and creatinine (correlation coefficients 0.198, 0.334, 0.104, 0.142, P<0.01), and lower levels of albumin (correlation coefficient 0.308, P<0.01). Multivariate logistic regression analysis showed that co-infection with Streptococcus pneumoniae or Mycoplasma was an independent risk factor for severe HRV pneumonia [ OR=1.611, 95% confidence interval ( CI):1.066-2.435, P<0.05; OR=3.355, 95% CI:2.062-5.458, P<0.01]. Conclusion:The infection rate of HRV is higher in preschool and school-age children. Severe HRV pneumonia is associated with increased levels of LDH, CRP, neutrophils, and creatinine, as well as decreased levels of albumin. Co-infection with Streptococcus pneumoniae or Mycoplasma may be an independent risk factor for severe HRV pneumonia. High-risk children require enhanced monitoring and early intervention to improve prognosis.

Objective To discuss the correlation between free prostate specific antigen(fPSA),serum ferritin(SF),blood uric acid(SUA)levels and Gleason grading in patients with prostate cancer(PCa).Methods The clinical data of 61 patients with prostate biopsy treated in 411 Hospital of Shanghai University from January to December of 2023 were retrospectively analyzed.According to the results of puncture,the patients were divided into benign prostatic hyperpla-sia(BPH)group(31 cases)and PCa group(30 cases).The levels of fPSA,SF and SUA in patients and Gleason grade in biopsy cases were analyzed.The correlation between fPSA,SF and SUA levels and Gleason grade was analyzed by the method of Spearman.And the diagnostic efficacy of fPSA,SF and SUA levels on PCa was analyzed by receiver operating characteristic curve(ROC).Results The levels of fPSA,SF and SUA in PCa group were significantly higher than those in BPH group(P＜0.05).There were statistically significant differences in levels of fPSA SF and SUA in PCA patients with different Gleason grades(P＜0.05).With the specificity reaching 96.08％and sensitivity reaching 94.35％,the ar-ea under the curve(AUC)of the combined fPSA,SF and SUA levels in the diagnosis of PCa was 0.982,which was higher than that of the single fPSA,SF and SUA levels(P＜0.05).SF and SUA levels in PCa patients were positively correlated with Gleason grade(P＜0.05),while fPSA levels were not correlated with Gleason grade(P＞0.05).Conclusion The levels of SF and SUA in PCa patients are positively correlated with Gleason grade,which can be used as an important index to predict Gleason grade in PCa patients.

To construct a recombinant fowl adenovirus 4(FAdV-4)expressing the Cap protein of goose astrovirus genotype 2(GoAstV-2),the expression cassette of Cap gene was inserted into the natural 1 966 bp deletion region of the FAdV-4 genome in the infectious clone p15A-cm-FAdV4-HNJZ.The resulted recombinant plasmid p15A-cm-FAdV4-HNJZ-Cap/GoAstV-2 was linearized with restriction enzyme and transfected into chicken hepatoma cell line(LMH)to rescue the recombinant FAdV-4 expressing the Cap protein of GoAstV-2,rF Ad V4-Cap/GoAstV-2.After 15 passages in LMH cells,the recombinant rFAdV4-Cap/GoAstV-2 was identified by PCR using primers flanking the insertion site of the Cap gene expression cassette and using viral genome DNA extracted from rFAdV4-Cap/GoAstV-2 infected LMH cells as template.LMH cells were in-fected with 15th passage rFAdV4-Cap/GoAstV-2 and indirect immunofluorescence was performed with a polyclonal antibody against Cap protein as the primary antibody.Western blot was carried out with lysates of rFAdV4-Cap/GoAstV-2 infected LMH cells.The in vitro replication dynamic of the 15th passage of the rFAdV4-Cap/GoAstV-2 was also investigated in LMH cells.The results demonstrated that the Cap gene of GoAstV-2 was presented in the genome of the recombinant vi-rus rF AdV4-Cap/Go Ast V-2,and could be expressed stably.The prepared recombinant virus in this study will lay a foundation for developing inactivated bivalent vaccine candidate against co-in-fection of FAdV-4 and GoAstV-2 in goose.

OBJECTIVE To evaluate the value of matrix-assisted laser desorption ionization time-of-flight-mass spectrometry(MALDI-TOF MS)in direct identification of carbapenem-resistant Escherichia coli and Klebsiella pneumoniae positive for blood culture.METHODS The blood culture bottles that were positive for E.coli or K.pneumoniae were collected from the patients with bloodstream infection who were treated in Zhongshan Hospi-tal,Fudan University from Jul.2021 to Jun.2023.The isolates were collected by using a gel-contact clotting tube,then the tested strains were respectively mixed with 4 μg/ml of imipenem,4 μg/ml of meropenem and 2 μg/ml of ertapenem for coculture and were incubated at 35 ℃ for 4 and 5 hours,finally,the strains were i-dentified by using the mass spectrum.The minimum inhibitory concentrations(MICs)of the three types of drugs were tested by microbroth dilution method and were set as the golden standards for the test.RESULTS Totally 31 strains of E.coli and 28 strains of K.pneumoniae that were positive in blood culture bottles were collected,both of the effective rates of controlled growth of the E.coli strains were 100.00％after the incuba-tion for 4 and 5 hours,and the effective rates of controlled growth of the K.pneumoniae strains were 9 6.43％and 100.00％after the incubation for 4 and hours,respectively.The sensitivity,specificity,positive predictive value and negative predictive value of the K.pneumoniae strains against the three types of drugs were 100.00％after the incubation for 5 hours.The specificity and positive predictive value of the E.coli strains against imipenem,meropenem and ertapenem were 100.00％after the incubation for 5 hours;the sensitivities were 73.58％,78.93％and 78.93％,respectively;the negative predictive values were 70.60％,75.00％and 75.00％,respectively.CONCLUSIONS MALDI-TOF MS is rapid and accurate for direct identification of the carbapenem-resistant E.coli and K.pneumoniae strains positive in blood culture bottles,and the accuracy reaches at 100.00％for the test of drug-resistant K.pneumoniae strains after the incubation for 5 hours.The method may provide evidence for clinical treatment of bloodstream infections induced by carbapenem-resistant Enterobacteriaceae.

Objective:To identify the risk factors of venous thromboembolism (VTE) in postoperative renal transplantation recipients by Meta-analysis, and to provide evidence-based reference for clinical staff to develop early VTE prevention strategies.Methods:PubMed, Web of Science, Cochrane Library, Embase, China National Knowledge Infrastructure, VIP database, Wanfang database and Chinese Biomedical Literature Database were searched to collect the studies on the risk factors of postoperative VTE in kidney transplant recipients. The search period was from the establishment of the database to March 10, 2024. After literature screening, data extraction and quality evaluation were conducted independently by two researchers, Meta-analysis was performed using RevMan 5.3 software.Results:A total of 15 literatures with 20 influencing factors were included. Meta-analysis showed that age ( MD = 6.36, 95% CI 2.56-10.17, P<0.05), body mass index ( MD = 1.83, 95% CI 0.15-3.50, P<0.05), VTE history ( OR = 2.04, 95% CI 1.08-3.86, P<0.05), blood transfusion history ( OR = 3.77, 95% CI 2.43-5.83, P<0.05), glomerular filtration rate ( MD = -5.54, 95% CI -9.93 - -0.91, P<0.05), donor age ( MD = 3.18, 95% CI 1.10-5.25, P<0.05), combination of malignant tumor ( OR = 2.87, 95% CI 1.45-5.68, P<0.05), end-stage renal disease as polycystic kidney disease ( OR = 1.76, 95% CI 1.39-2.22, P<0.05), and interstitial nephritis ( OR = 1.60, 95% CI 1.06-2.40, P<0.05) were the influencing factors for postoperative VTE in renal transplant recipients. Conclusions:Clinical medical staff should actively identify high-risk groups for VTE after kidney transplantation by considering the 8 influencing factors determined by this study, and take targeted measures early to reduce the risk of postoperative VTE.

Objective:To systematically retrieve, extract and summarize the best evidence regarding the prevention and management of perioperative gastrointestinal dysfunction in patients with colorectal cancer, and to provide an evidence-based basis for clinical nursing practice.Methods:All evidence on the prevention and management of perioperative gastrointestinal dysfunction in colorectal cancer patients in various domestic and international databases and websites such as UpToDate, China national knowledge infrastructure, etc. were searched according to the "6S" pyramid model, including clinical decisions, guidelines, expert consensus, systematic evaluations, and randomized controlled trials from the time of database construction to August 15, 2024, on a top-to-bottom basis. Literature screening, quality assessment, and evidence extraction were performed independently by 2 nursing researchers.Results:A total of 19 documents were finally included, included 4 guidelines, 3 expert consensus, 7 systematic evaluations, 1 Meta-analysis, and 4 randomized controlled trials, and 35 pieces of evidence were summarized in 6 areas, including screening and assessment, build a multidisciplinary team, preoperative preparation, intraoperative monitoring, postoperative care, and evaluation of outcomes.Conclusions:The evidence summarized in this study is both scientifically rigorous and practically applicable, and can be used by healthcare professionals to design care plans for colorectal cancer patients that accelerate the recovery of gastrointestinal function, thereby promoting evidence-based clinical nursing practice.

OBJECTIVE To provide references for further improving the management level of Chinese herbal pieces covered by the healthcare security drug catalog(hereinafter referred to as"healthcare security covered Chinese herbal pieces").METHODS Policy documents related to healthcare security covered Chinese herbal pieces were retrieved from official websites of National Healthcare Security Administration and medical insurance bureaus of various provinces(autonomous regions,municipalities directly under the central government).Using text analysis,the current management status of healthcare security covered Chinese herbal pieces in various provinces was analyzed from two dimensions:management regulations and catalog formats.RESULTS In terms of the management regulations for healthcare security covered Chinese herbal pieces,some provinces implemented both national and provincial standards for Chinese herbal pieces,while a small number of provinces adopted national standards alongside their own provincial standards.Regarding the payment categories of healthcare security covered Chinese herbal pieces,most were classified as category A,though some provinces managed them as category B,and others included both categories A and B or category C.In terms of applicants,there were three kinds:only medical institutions,only herbal piece enterprises,or both medical institutions and herbal piece enterprises being allowed to apply.During the application process,some provinces required preliminary evaluations by local healthcare security departments with recommended lists submitted,while some provinces omitted preliminary evaluations and relied solely on unified evaluation by provincial healthcare security departments.Regarding the methods for catalog formulation and adjustment,there was little variation among the provinces.In terms of the catalog format for healthcare security covered Chinese herbal pieces,some provinces had supplemented the catalog fields based on the national medical insurance catalog.In contrast,some provinces had relatively simple catalogs.Regarding the sorting of the herbal pieces catalog,some catalogs were arranged by the number of Chinese character strokes in the names of the herbal pieces,but most catalogs lacked clear sorting rules.CONCLUSIONS Significant results have been achieved in formulating management measures and the catalog of healthcare security covered Chinese herbal pieces.However,there is still a great necessity to develop targeted and adaptable management clauses based on local conditions and the characteristics of Chinese herbal pieces.The content and format of healthcare security drug catalog of Chinese herbal pieces should be improved;in addition,continuous tracking of the latest management policies is also essential to improve the accuracy and operability of formulated catalog.