AIM:To investigate the effects of high concentration of glucose on the expressions of serum and glucocorticoid induced protein kinase 1 (SGK1) and fibronectin in human podocytes and detect the production of fibronectin in human podocytes transfected with active and mutant forms of SGK1.METHODS: The expressions of SGK1 and fibronectin in human podocytes exposed in high glucose for 24 h were detected by using Western blotting. Human podocytes were transfected with three kinds of plasmid: pIRES2-EGFP-SGK1SD (SGK1-active plasmid), pIRES2-EGFP-SGK1KN (SGK1-mutant plasmid) and pIRES2-EGFP (empty vector plasmide). The synthesis levels of fibronectin were detected by immunofluorescence.RESULTS: The expression of SGK1 in normal human podocytes was observed and the expression level was up-regulated in podocytes exposed to high glucose for 24 h (50?4 vs 35?3), parallelly with the high expression of fibronectin (19?4 vs 12?2). The secretion of fibronectin was increased obviously in SGK1-active plasmid (SGK1SD) transfected podocytes compared with the vector transfected podocytes. Whereas, the fibronectin level was decreased significantly but not completely abolished in SGK1-mutant plasmid (SGK1KN) transfected podocytes, compared with SGK1SD transfected podocytes. CONCLUSION: SGK1 may be involved in the synthesis of fibronectin in human podocytes induced by high glucose and play a critical role in the activation of human podocytes during early stage of diabetic nephropathy.

Objective To analyze the value of conventional T2WI combining with breath-hold Cartesian VIBE sequence, and compared with Blade combining with breath-free radial VIBE sequences in pre-operative T staging of potentiallly resectable esophageal carcinoma. Methods Sixty-five cases of esophageal carcinoma were concluded prospectively. All patients had received pathological examination of gastroscope biopsy before MRI. Patients received MR examination, including T2WI, breath-hold VIBE, Blade, and breath-free radial VIBE sequences. Two radiologists with more than five years experiences in the diagnosis of chest, performed T staging in MRIby referring to the the 7th edition of UICC-AJCC TNM classification. The results of MRI T staging and the postoperative pathological T staging were analyzed byχ2 test. Results Sixty-five patients were included. Diagnostic coincidence rates of the preoperative T staging by using conventional T2WI combining with breath-hold Cartesian VIBE sequences and Blade combining with breath-free radial VIBE sequences were 51%(33/65) and 88%(57/65) ,with 32 and 8 cases overstaging or understaging respectively, and the statistical differences were significant(χ2=20.80, P<0.05). The former diagnostic accuracy of esophageal carcinoma in situ, muscularis violation and esophageal fiber membrane were 42%(8/19), 54%(14/26), 55%(11/20), and the latter were 89%(17/19), 88%(23/26), and 85%(17/20). Conclusions Diagnostic coincidence rate of the preoperative T staging by using Blade combining with breath-free radial VIBE sequences is much higher than conventional T2WI combining with breath-hold Cartesian VIBE sequences. Blade combining with breath-free radial VIBE sequences could be used as non-invasive imaging method in preoperative T staging of potentially resectable esophageal carcinoma.

Acute kidney injury (AKI) is often associated with organ donation and renal transplantation, which leads to an increase of fatality rate, hospitalization time and hospitalization costs. In recent years, studies have shown that ferroptosis is closely related to AKI, but the exact molecular biological mechanism has not been clarified, which need more research. In this article, the role of ferroptosis in AKI was reviewed from the aspects of ferroptosis related biomarkers and biological reactions, in order to find a new possible direction for the prevention and treatmentof AKI.

Objective:To explore the prognostic utility of LifePort perfusion parameters plus perfusate biomarkers for predicting delayed graft function(DGF)and recovery time during deceased donor kidney transplantation(KT).Methods:From January 1, 2019 to August 31, 2019, retrospective analysis was performed for clinical data of 113 KT recipients. Based upon whether or not DGF occurred within 3 months, they were divided into two groups of DGF group(20 cases)and non-DGF (93 cases). Two groups were compared using LifePort perfusion parameters, biomarker concentrations, incidence of DGF and kidney recovery time. Statistical analysis was performed.Results:The incidence of DGF was 17.7%(20/113); Multivariate Logistic regression results indicated that terminal resistance(OR 1.879, 95% CI 1.145~3.56)and glutathione S-transferase(GST)(OR 1.62, 95% CI 1.23~2.46)were independent risk factors for DGF; Cox hazard model revealed that terminal resistance was a risk factor for recovery time of renal function(HR=0.823, 95% CI 0.735~0.981). The model combining terminal resistance and GST(AUC=0.888, 95% CI 0.842~0.933)significantly improved the predictive efficacy for DGF as compared with using terminal resistance(AUC=0.756, 95% CI 0.693~0.818)or GST alone(AUC=0.729, 95% CI 0.591~0.806).Conclusions:Combining LifePort perfusion parameters and fluid biomarkers can improve the predictive utility of DGF.

Objective To analyze the prediction efficiency of scoring models at home and abroad on delayed graft function (DGF) after renal transplantation in China. Methods The clinical data of 112 donors and 220 recipients undergoing renal transplantation were prospectively analyzed. The DGF predicted by KDRI model, Jeldres model, and model of our center was compared with actual DGF incidence of renal transplant recipients. The prediction efficiency of each model was analyzed. The predictive accuracy was compared by the area under curve (AUC) of receiver operating characteristic (ROC) curve. Results The DGF incidence of 220 renal transplant recipients was 14.1% (31/220). DGF prediction using KDRI model showed that 41 cases were high risk donors, the AUC was 0.57, the sensitivity was 0.37, the specificity was 0.66, and the positive predictive value was 22%. DGF prediction using Jedres model showed that 22 cases were high risk recipients, the AUC was 0.56, the sensitivity was 0.13, the specificity was 0.92 and the positive predictive value was 20%. DGF prediction using the model of our center showed that 25 cases were high risk donors, the AUC was 0.80, the sensitivity was 0.53, the specificity was 0.84, the positive predictive value was 40%. Conclusions Compared with the KDRI and Jedres models, the prediction model of our center has higher AUC and sensitivity with a better prediction efficiency on DGF. Therefore, it is a suitable evaluation system of donors from donation after citizen's death in Chinese.

Objective:To investigate the relevant situation of the emergency departments (ED) of general hospitals at all levels in Zhejiang Province participating in the prevention and control of Corona Virus Disease 2019 (COVID-19), and to provide a reference for further improving the capacity of emergency services.Methods:A multi-center cross-sectional investigation study was jointly initiated by the Zhejiang Provincial Emergency Medicine Quality Control Center, the Emergency Medicine Branch of Zhejiang Medical Association, and the Emergency Physicians Branch of Zhejiang Medical Doctors Association. Before investigation, experts were organized to develop a special questionnaire. Then the ED of general people's hospitals or Chinese Medicine hospitals at the county/district-level or above were included and their participations in the prevention and control of COVID-19 epidemic between March 9, 2020 and March 15, 2020 were investigated. The collected information included: basic information of the hospitals; participation of ED in fever clinics; the number of COVID-19 patients admitted to ED; the situation of ED support to the anti-epidemic front-line; and the epidemic prevention facilities of ED, and the early supply of medical protective equipments.Results:A total of 181 hospitals from 11 prefectures and cities across the province were finally included, including 52 (28.7%) Grade-A tertiary hospitals, 45 (24.9%) Grade-B tertiary hospitals, 64(35.4%) Grade-A secondary hospitals, and 20 (11.0%) Grade-B secondary or lower hospitals. Among them, 93 (51.4%) were COVID-19 designated hospitals, and 49 (27.1%) had the ability to detect COVID-19 nucleic acid; 177 (97.8%) set up independent fever clinics and 33 (18.6%) were managed by ED alone or mainly. The medical staffs of 65 (36.7%) fever clinics were all or mainly from ED. A total of 213 cases of COVID-19 were received/treated in 40.3% ED, accounting for 17.3% of the total number of cases in Zhejiang Province. A total of 3 848 doctors and nurses supported the front-line of epidemic prevention, of them, 253 supported Hubei Province (accounting for 12.5% ??of the total number). There were 68.5% and 76.8% ED having spare separate clinic and ordinary rescue single room to deal with the COVID-19. In addition, at the early days of anti-epidemic, more than 50% ED were in short supply or even had no medical protective equipment.Conclusions:The ED of Zhejiang Province plays an important role and has made a great contribution to the prevention and control of the COVID-19 epidemic. It is necessary to improve the conditions of ED to effectively respond to the prevention and control of daily infectious diseases.

BACKGROUND: Delayed graft function (DGF) is the main cause of renal function failure after kidney transplantation. This study aims at investigating the value of hypothermic machine perfusion (HMP) parameters combined with perfusate biomarkers on predicting DGF and the time of renal function recovery after deceased donor (DD) kidney transplantation. METHODS: HMP parameters, perfusate biomarkers and baseline characteristics of 113 DD kidney transplantations from January 1, 2019 to August 31, 2019 in the First Affiliated Hospital of Xi'an Jiaotong University were retrospectively analyzed using univariate and multivariate logistic regression analysis. RESULTS: In this study, the DGF incidence was 17.7% (20/113); The multivariate logistic regression results showed that terminal resistance (OR: 1.879, 95% CI 1.145-3.56) and glutathione S-transferase (GST)(OR = 1.62, 95% CI 1.23-2.46) were risk factors for DGF; The Cox model analysis indicated that terminal resistance was an independent hazard factor for renal function recovery time (HR = 0.823, 95% CI 0.735-0.981). The model combining terminal resistance and GST (AUC = 0.888, 95% CI: 0.842-0.933) significantly improved the DGF predictability compared with the use of terminal resistance (AUC = 0.756, 95% CI 0.693-0.818) or GST alone (AUC = 0.729, 95% CI 0.591-0.806). CONCLUSION According to the factors analyzed in this study, the combination of HMP parameters and perfusate biomarkers displays a potent DGF predictive value.
Biomarkers ; Delayed Graft Function ; Graft Survival ; Humans ; Kidney/physiology* ; Kidney Transplantation/adverse effects* ; Organ Preservation ; Perfusion ; Retrospective Studies ; Tissue Donors

AIM: To evaluate the binocular visual function in high myopia patients after the implantation of implantable collamer lens(ICL)V4c.METHODS: A total of 35 cases(70 eyes)that received binocular ICL implantation at our hospital from May 2019 to May 2021 were enrolled in this prospective study. Binocular full-range visual acuity, contrast sensitivity, stereopsis, mesopic vision and glare sensitivity, and monocular wavefront and the quality of vision questionnaire were assessed before the surgery and at 1 mo postoperatively.RESULTS: At 1 mo postoperatively, 35 cases(100%)had binocular uncorrected distance visual acuity(UDVA)≤0.00(LogMAR), 16 cases(46%)had binocular UDVA≥preoperative corrected distance visual acuity(CDVA). Binocular UDVA and uncorrected intermediate visual acuity(UIVA,80 cm)were improved compared to preoperative CDVA and distance-corrected intermediate visual acuity(DCIVA,80 cm)(all P<0.05).While there were no differences in the binocular postoperative UIVA(60 cm)and preoperative DCIVA(60 cm),and uncorrected near visual acuity(UNVA,40 cm)and preoperative distance-corrected near visual acuity(DCNVA,40 cm)(all P>0.05). The binocular contrast sensitivity was significantly improved postoperatively(P=0.001), and the postoperative binocular mesopic vision, glare sensitivity(no glare/glare)and binocular stereopsis(5 m/40 cm)had no differences(all P>0.05). The postoperative total higher-order aberration, trefoil aberration, coma and spherical aberration were increased, besides the median of total coma in the right eye with a pupil diameter of 3.0 mm was decreased after surgery. The mean total score of quality of vision questionnaire was significantly increased from 54.87 preoperatively to 80.92 after implantation(P<0.05), with high satisfaction and no obvious visual disturbance in patients.CONCLUSION: Although the monocular high-order aberrations increased in the early stage after ICL V4c binocular implantation in patients with high myopia, the binocular visual function was improved.

Targeted point mutagenesis through homologous recombination has been widely used in genetic studies and holds considerable promise for repairing disease-causing mutations in patients. However, problems such as mosaicism and low mutagenesis efficiency continue to pose challenges to clinical application of such approaches. Recently, a base editor (BE) system built on cytidine (C) deaminase and CRISPR/Cas9 technology was developed as an alternative method for targeted point mutagenesis in plant, yeast, and human cells. Base editors convert C in the deamination window to thymidine (T) efficiently, however, it remains unclear whether targeted base editing in mouse embryos is feasible. In this report, we generated a modified high-fidelity version of base editor 2 (HF2-BE2), and investigated its base editing efficacy in mouse embryos. We found that HF2-BE2 could convert C to T efficiently, with up to 100% biallelic mutation efficiency in mouse embryos. Unlike BE3, HF2-BE2 could convert C to T on both the target and non-target strand, expanding the editing scope of base editors. Surprisingly, we found HF2-BE2 could also deaminate C that was proximal to the gRNA-binding region. Taken together, our work demonstrates the feasibility of generating point mutations in mouse by base editing, and underscores the need to carefully optimize base editing systems in order to eliminate proximal-site deamination.
APOBEC-1 Deaminase ; genetics ; metabolism ; Animals ; Bacterial Proteins ; genetics ; metabolism ; Base Sequence ; CRISPR-Associated Protein 9 ; CRISPR-Cas Systems ; Cytidine ; genetics ; metabolism ; Embryo Transfer ; Embryo, Mammalian ; Endonucleases ; genetics ; metabolism ; Gene Editing ; methods ; HEK293 Cells ; High-Throughput Nucleotide Sequencing ; Humans ; Mice ; Mice, Inbred C57BL ; Microinjections ; Plasmids ; chemistry ; metabolism ; Point Mutation ; RNA, Guide ; genetics ; metabolism ; Thymidine ; genetics ; metabolism ; Zygote ; growth & development ; metabolism ; transplantation

Genome editing tools such as the clustered regularly interspaced short palindromic repeat (CRISPR)-associated system (Cas) have been widely used to modify genes in model systems including animal zygotes and human cells, and hold tremendous promise for both basic research and clinical applications. To date, a serious knowledge gap remains in our understanding of DNA repair mechanisms in human early embryos, and in the efficiency and potential off-target effects of using technologies such as CRISPR/Cas9 in human pre-implantation embryos. In this report, we used tripronuclear (3PN) zygotes to further investigate CRISPR/Cas9-mediated gene editing in human cells. We found that CRISPR/Cas9 could effectively cleave the endogenous β-globin gene (HBB). However, the efficiency of homologous recombination directed repair (HDR) of HBB was low and the edited embryos were mosaic. Off-target cleavage was also apparent in these 3PN zygotes as revealed by the T7E1 assay and whole-exome sequencing. Furthermore, the endogenous delta-globin gene (HBD), which is homologous to HBB, competed with exogenous donor oligos to act as the repair template, leading to untoward mutations. Our data also indicated that repair of the HBB locus in these embryos occurred preferentially through the non-crossover HDR pathway. Taken together, our work highlights the pressing need to further improve the fidelity and specificity of the CRISPR/Cas9 platform, a prerequisite for any clinical applications of CRSIPR/Cas9-mediated editing.
Blastocyst ; CRISPR-Cas Systems ; Hemoglobins, Abnormal ; genetics ; metabolism ; Humans ; Zygote