To explore the application effect of host restriction factors(HRFs)in the development of antiviral gene drugs,this study based on the"common pathway of viral infection"and"host innate immunity HRFs",the genes of three antiviral HRFs,namely cholesterol-25-hydroxylase(CH25H),interferon-induced transmembrane protein(IFITM3)and interferon-stimulated gene 15(ISG15)were fused and expressed using pCK vectors to construct antiviral gene drugs.The fusion expression gene sequence CH25H-IFITM3-ISG15(C Ⅱ)was constructed by linking the gene cod-ing sequences of these three HRFs through the cleavage peptide coding sequence.Subsequently,the C Ⅱ sequence was amplified by PCR,ligated to the pCK expression vector,and the recombinant plasmid was transformed,identified,and extracted to obtain a candidate biodrug based on the DNA expression system,which was named pCK-CⅡ.Then,the recombinant plasmid was transfected in-to HEK 293T cells,and the expression of three antiviral proteins was successfully detected by Western blot.To clarify the antiviral effect of pCK-C Ⅱ at the cellular level,pCK-C Ⅱ was trans-fected into HEK 293 cells and BHK-21 cells,respectively.Twenty-four hours later,the BHK-21 cells were infected with vesicular stomatitis virus expressing green fluorescent protein(VSV-GFP),12 h later,the cells were observed under a fluorescence microscope and detected by flow cy-tometry;HEK 293 cells were inoculated with H3N2 subtype influenza virus,and the expression of H3N2 subtype influenza virus nuclear proteins was detected after 12 h using Western blot.The re-sults showed that transient transfection of pCK-C Ⅱ plasmid could significantly reduce the fluores-cence level of cells and the expression of nuclear protein of H3N2 subtype influenza virus in infec-ted cells.These results indicated that pCK-C Ⅱ had an inhibitory effect on the infection of VSV-GFP and H3N2 subtypes of influenza viruses after transient transfection of cells.

To explore the application effect of host restriction factors(HRFs)in the development of antiviral gene drugs,this study based on the"common pathway of viral infection"and"host innate immunity HRFs",the genes of three antiviral HRFs,namely cholesterol-25-hydroxylase(CH25H),interferon-induced transmembrane protein(IFITM3)and interferon-stimulated gene 15(ISG15)were fused and expressed using pCK vectors to construct antiviral gene drugs.The fusion expression gene sequence CH25H-IFITM3-ISG15(C Ⅱ)was constructed by linking the gene cod-ing sequences of these three HRFs through the cleavage peptide coding sequence.Subsequently,the C Ⅱ sequence was amplified by PCR,ligated to the pCK expression vector,and the recombinant plasmid was transformed,identified,and extracted to obtain a candidate biodrug based on the DNA expression system,which was named pCK-CⅡ.Then,the recombinant plasmid was transfected in-to HEK 293T cells,and the expression of three antiviral proteins was successfully detected by Western blot.To clarify the antiviral effect of pCK-C Ⅱ at the cellular level,pCK-C Ⅱ was trans-fected into HEK 293 cells and BHK-21 cells,respectively.Twenty-four hours later,the BHK-21 cells were infected with vesicular stomatitis virus expressing green fluorescent protein(VSV-GFP),12 h later,the cells were observed under a fluorescence microscope and detected by flow cy-tometry;HEK 293 cells were inoculated with H3N2 subtype influenza virus,and the expression of H3N2 subtype influenza virus nuclear proteins was detected after 12 h using Western blot.The re-sults showed that transient transfection of pCK-C Ⅱ plasmid could significantly reduce the fluores-cence level of cells and the expression of nuclear protein of H3N2 subtype influenza virus in infec-ted cells.These results indicated that pCK-C Ⅱ had an inhibitory effect on the infection of VSV-GFP and H3N2 subtypes of influenza viruses after transient transfection of cells.

Objective To investigate the expression levels of serum insulin-like growth factor binding protein-7 (IGFBP-7) and silence-information regulatory factor 4 (SIRT4) in patients with acute heart failure (AHF) and their prognostic value. Methods A total of 151 patients with AHF (AHF group) and 151 healthy subjects (control group) were enrolled. Serum IGFBP-7, SIRT4, N-terminal natriuretic peptide precursors (NT-proBNP) and reactive oxygen species (ROS) were detected in the two groups. The relationships of serum levels of IGFBP-7, SIRT4, NT-proBNP, ROS with clinical grade were analyzed. Pearson correlation coefficient was used to analyze the correlations of IGFBP-7 and SIRT4 with NT-proBNP and ROS. Multivariate Logistic regression analysis was used to analyze the prognostic factors of AHF patients, and the Receiver operating characteristic (ROC) curve was drawn to analyze the predictive efficacy of serum IGFBP-7 and SIRT4 on poor prognosis of AHF patients. Results Compared with the control group, the expression levels of IGFBP-7, SIRT4, NT-proBNP and ROS in serum of the AHF group were significantly increased (P < 0.05). The expression levels of IGFBP-7, SIRT4, NT-proBNP and ROS in the serum of patients with clinical disease grade Ⅳ were higher than that in grade Ⅲ and grade Ⅱ, and those with grade Ⅲ was significantly higher than that of grade Ⅱ (P < 0.05). The expression levels of IGFBP-7, SIRT4, NT-proBNP and ROS and severity grading in patients with poor prognosis in the AHF group were significantly higher than those in patients with good prognosis (P < 0.05). The expression levels of IGFBP-7 and SIRT4 in serum were positively correlated with the levels of NT-proBNP and ROS (r=0.523, 0.498, 0.578, 0.557, P < 0.05). Multifactor Logistic regression analysis showed that clinical grade, serum expression levels of IGFBP-7, SIRT4, NT-proBNP and ROS were independent influencing factors for prognosis of AHF (P < 0.05). ROC curve analysis showed that serum IGFBP-7 and SIRT4 had a certain predictive value for prognosis in AHF patients, with AUC of 0.794 and 0.795, and the combined detection of IGFBP-7 and SIRT4 had a higher predictive value (AUC was 0.909, 95%CI, 0.858 to 0.959). Conclusion Serum IGFBP-7 and SIRT4 are highly expressed in patients with AHF, and their expression levels are correlated with clinical grade and prognosis, and the combined detection of the two indicators is of high reference value for evaluating the prognosis of patients.

Objective To investigate the correlation between the total CSVD burden and fundus vasculopathy,and evaluate significance of fundus vasculopathy for cognitive impairment in CSVD patients.Methods A total of 290 inpatients who taking physical examination in our department from May 2021 to August 2022 were consecutively recruited,and according to their score of CSVD,they were divided into group 0(129 cases),group 1(51 case),group 2(42 cases),group 3(44 cases),and group 4(24 cases).All the subjects underwent brain magnetic resonance imaging,fundus fluorescein angiography,and cognitive function assessment.The total CSVD burden and fundus vasculopathy were evaluated.The general clinical data,results of laboratory tests,fundus vascularization,and cognitive function were compared among the groups with different CSVD burden scores.Spearman correlation analysis and linear correlation analysis were used to explore the correlation between total CSVD burden score and fundus vascular disease.Results Significant differences were observed in terms of age,years of education,cerebral infarction/TIA,total cho-lesterol,LDL-C,creatinine,MoC A and MMSE scores,positive results of connectivity test,digit-symbol conversion test,Stroop colour-word interference test and verbal fluency test,values of CRAE,CRVE,AVR,Scheie grade,DWMH and PVWMH,enlarged perivascular space in the basal ganglia(BG-EPVS),lacunar infarct and cerebral microbleeds(CMB)in different CSVD total bur-den groups(P＜0.05,P＜0.01).Spearman correlation analysis showed that total CSVD burden was negatively correlated with CRAE and AVR(r=-0.655,P=0.000;r=-0.679,P=0.000),and positively with CRVE and Scheie grade(r=0.560,P=0.000;r=0.685,P=0.000).Multivari-ate linear analysis showed that the total CSVD burden after adjusting for relevant risk factors was significantly correlated with CRAE,CRVE,AVR and Scheie grades(P＜0.01).Conclusion Fun-dus vasculopathy is strongly associated with increased total CSVD burden,and it can be regarded as a valid predictor of CSVD-related cognitive impairment.

Objective:To study the growth trajectory and the incidence of malnutrition and clinical events in infants with congenital heart disease in 1 year after surgery.Methods:Children at the age of 1 year or younger who were diagnosed with congenital heart disease and underwent surgery at Guangzhou Women and Children's Medical Center affiliated to Guangzhou Medical University from January 2018 to January 2019 were included. The age, gender, birth weight and length, and parental height and weight were collected, and the occurrence of clinical events of interest and the children's health as evaluated by caregiver within 1 year after surgery were followed up through questionnaire survey. Malnutrition was defined as a weight-for-age z-score (WAZ) ≤-2 at 1 year after surgery and a WAZ>-2 was defined as non-malnourished.Results:Among the 502 children, 301 were boys and 201 were girls, aged 4.1 (range: 2.0 to 6.8) months, of whom 64.7% were with simple congenital heart disease and 35.3% complex congenital heart disease. The children were generally with mild malnutrition (WAZ<-1 and >-2) before surgery. At 3 months and 6 months after surgery, the children showed a rapid growth catch-up, although failing to reach the normal level, and the trend plateaued at 1 year after surgery. The proportion of children with malnutrition decreased gradually within 1 year after surgery. 47.0% of included children had malnutrition before surgery, and the proportion decreased significantly at 3 months and 6 months, to 17.9% at 1 year after surgery. After discharge, these children suffered from upper respiratory infection most commonly (74.5%), followed by pneumonia (41.2%) and diarrhea (36.7%), and vomiting and constipation (22.1%). In terms of children's health status as evaluated by parents, about 32.0% of families considered their children in poor health before surgery, and the proportion decreased to 6.9% within 1 year after surgery.Conclusion:Infants with congenital heart disease continued to grow and catch up within 1 year after surgery, showing significantly improved nutrition status, but some children still experienced malnutrition at 1 year after surgery.

Objective:This study aimed to explore the performance of renal resistive index (RRI), semiquantitative power Doppler ultrasound (PDU) score, and renal venous Doppler waveform (RVDW) pattern in predicting 28-day renal dysfunction in critically ill patients and establish nomogram model.Methods:This was a prospective, observational study. Critically ill patients admitted to the emergency intensive care unit (ICU) of Cangzhou Central Hospital from January 2018 to October 2022 were included. Patients underwent renal ultrasound examination to obtain RRI, PDU score and RVDW pattern within 24 h after ICU admission. The following clinical variables were collected during the renal ultrasound examination session, including heart rate, mean arterial pressure, type and dose of vasoactive drugs, oxygen therapy parameters, and average urine volume per hour derived from a period of 6 h prior to the ultrasound examination. The data on duration of AKI and mortality were recorded on the 28th day of follow-up. Patients were divided into 28-day normal renal function group and 28-day renal dysfunction group according to 28-day renal dysfunction. 28-days of renal dysfunction was defined as failure to achieve renal function recovery within 28 days of ICU admission. The difference of each index between the two groups was compared. Associated factors for 28-day renal dysfunction were determined by univariate and multivariate COX regression analyses. A nomogram was developed based on the independently factors associated with 28-day renal dysfunction. Survival receiver operator characteristic (ROC) curves were plotted to assess diagnostic performance in predicting 28-day renal dysfunction. Delong’s test was used to compare area under the curves (AUC) between each predictor.Results:187 patients were enrolled for the final analysis: 97 with no AKI, 48 with AKI stage 1, 24 with AKI stage 2, and 18 with AKI stage 3 upon enrollment. At 28-day follow up, 16 patients had renal dysfunction and 2 required continuous renal replacement therapy (CRRT). The multivariate COX regression showed that RVDW and SCr upon enrollment were the independent risk predictors. Nomogram based on RVDW and SCr upon enrollment showed the best performance in predicting 14-day renal dysfunction (AUC = 0.918, 95% CI:0.871-0.964, P<0.05), and the AUC was statistically significantly higher than single index (all P<0.05). Nomogram also showed the best performance in predicting 28-day renal dysfunction (AUC = 0.924, 95% CI:0.865-0.983, P<0.05), and the AUC was statistically significantly higher than single index (all P<0.05) except for SCr upon enrollment. The optimal cutoff for nomogram in predicting 28-day renal dysfunction was ≤89.5 (sensitivity, 81.2%; specificity, 90.6%; Youden index, 0.719). Kaplan-Meier analysis showed that the median duration of renal dysfunction in the groups with total nomogram score >85.9 and ≤85.9 was 0 and 22 days (HR=0.220, 95% CI:0.129-0.376, P<0.001). Conclusions:SCr and RVDW pattern within 24 h from ICU admission were independent factors associated with 28-day renal dysfunction in critically ill patients. The value of the nomogram model based on these two factors in predicting 28-day renal dysfunction is superior to each single intrarenal Doppler spectrum indicator and clinical indicator.

Objective:To retrieve, evaluate and summarize the relevant evidence on the management of hyperphosphatemia in adult maintenance hemodialysis patients.Methods:Domestic and foreign databases and relevant professional websites were searched for best practices, evidence summaries, guidelines, expert consensus and systematic reviews on hyperphosphatemia in adult maintenance hemodialysis patients. The retrieval time was from the establishment of the databases to January 2023. The methodological quality of the included articles was evaluated, and the best evidence was extracted and summarized.Results:A total of 13 articles were included, including 2 clinical decisions, 4 guidelines and 7 systematic reviews. A total of 23 evidences were summarized from 5 aspects, including blood phosphorus monitoring, adequate dialysis, diet control, drug management and health education.Conclusions:This study summarizes the best evidence on the management of hyperphosphatemia in maintenance hemodialysis patients and provides a reference for clinicians to manage hyperphosphatemia in adult maintenance hemodialysis patients.

Objective:To explore the predictive value of bedside ultrasound monitoring of optic nerve sheath diameter (ONSD) for short-term neurological prognosis in critically ill patients without primary brain injury.Methods:An observational prospective study was conducted to enroll critically ill patients without primary brain injury admitted to the emergency intensive care unit (ICU) of Cangzhou Central Hospital from January 2021 to April 2022. The exclusion criteria were as follows: age < 18, combined ocular and optic nerve pathology or injuries, impaired consciousness due to prior neuropathy, primary brain injury, ICU stay < 3 days, death or loss of follow-up within 28 days. Bedside ultrasound measurements of ONSD were performed within 24 hours of ICU admission and on day 3 of ICU admission. The consciousness status was assessed daily during ICU hospitalization. If the Glasgow Coma Scale (GCS) is 15 and the confusion assessment method intensive care unit (CAM-ICU) is negative, the consciousness status will be defined as nonconsciousness disorder. While if the GCS score is less than 15 or the CAM-ICU is positive, the consciousness status will be defined as consciousness disorder. According to the status of consciousness at 28 days, patients were divided into a nonconscious disorder group and a conscious disorder group, and the difference in each index was compared between the two groups. Univariate and multivariate Cox regression were used to analyze the factors influencing 28-day neurological function prognosis, and a Kaplan?Meier survival curve was plotted to analyze the relationship between ONSD and 28-day neurological function prognosis.Results:Sixty-one critically ill patients without primary brain injury (48 in the nonconscious disorder group and 13 in the conscious disorder group) were recruited. Compared to patients in the unconscious disorder group, those in the conscious disorder group had lower GCS upon ICU admission [7(4, 8) vs. 8(6, 14), P<0.05], longer length of mechanical ventilation (MV) [28(15, 28) days vs. 10(4, 14) days, P<0.001], and longer length of ICU stay [28(28, 28) days vs. 12(7, 20) days, P<0.001]. Patients in the conscious disorder group had a higher ONSD within 24 hours of ICU admission [(5.75±0.53) mm vs. (5.45±0.60) mm, P=0.114] and a higher ONSD 3 days after ICU admission [(5.54±0.64) mm vs. (5.22±0.65) mm, P=0.124] than patients in the unconscious disorder group, but the differences were not statistically significant. Multivariate Cox regression analysis showed that use of MV, GCS upon ICU admission and ONSD on day 3 of ICU admission were independent risk factors. Kaplan?Meier survival analysis showed that patients with an ONSD < 5.30 mm on day 3 had a better 28-day neurological prognosis. Moreover, among the patients with ONSD within 24 hours ≥5.30 mm, the patients with ONSD decreased to < 5.30 mm on day 3 had significantly better 28-day neurological prognosis than those with ONSD ≥ 5.30 mm on day 3 ( P=0.042). Conclusions:ONSD within 24 hours of ICU admission, especially ONSD levels and changes in ONSD on day 3, had predictive value for the short-term neurological prognosis of critically ill patients without primary brain injury.

Objective:To compare the predictive ability of SpO 2/FiO 2(S/F) and ROX index on the failure of high-flow nasal cannula(HFNC)therapy in children with acute respiratory failure after congenital heart disease surgery, and to identify the best cut-off point. Methods:Through a case-control study, the clinical data of 371 children with acute respiratory failure after congenital heart surgery treated with HFNC admitted to Guangzhou Women and Children′s Medical Center from January 2018 to December 2021 were retrospectively analyzed.The primary outcome was the need for re-intubation within 48 h after extubation of invasive ventilation.The ability of S/F and ROX index to predict HFNC failure was compared, and the optimal cut-off point was determined based on the area under the curve (AUC) of receiver operating characteristic curve.Results:A total of 371 children were included, of whom 27 (7.3%) eventually required mechanical ventilation within 48 h. The S/F prediction accuracy was highest after 6 h of HFNC treatment(AUC=0.712, 95% CI 0.599-0.825, P=0.001), and the best cut-off point for S/F was 178 mmHg(1 mmHg=0.133 kPa)(sensitivity 74.9%, specificity 69.6%). Whereas the prediction accuracy of the ROX index was highest after 12 hours of HFNC treatment, the AUC was 0.737(95% CI 0.623-0.851, P=0.002), and the best cut-off point of the ROX index was 5.865(sensitivity 72.4% specificity 66.7%). The difference in AUC between S/F after 6 h of HFNC treatment and ROX after 12 h was not statistically significant ( P=0.444), with higher sensitivity and specificity, and earlier prediction time(6 hours) in the former. Conclusion:Children with acute respiratory failure after congenital heart surgery have a strong predictive ability of S/F after 6 h of HFNC treatment, and the risk of HFNC treatment failure is higher in children with S/F <178 mmHg.

Objective:To summarize the results of surgical treatment for complete atrioventricular septal defect(CAVSD) in early and middle stages.Methods:147 children with CAVSD in Guangzhou Women and Children’s Medical Center from January 2010 to December 2019 were selected, Males 85, females 62, median age of surgery 5 months(1 months-10 years old), median body mass 5.5 kg(2.4-20.9 kg). Complete atrioventricular septal defect was diagnosed by ultrasonic cardiogram before surgery. All the children underwent atrial ventricular valve formation and underwent simultaneous repair.Outpatient follow-up was planned.Ultrasonic cardiogram and electrocardiogram were performed. SPSS 22 statistical software was used for data analysis.Results:All 147 CAVSD patients underwent one-time surgical correction.Early postoperative death occurred in 7 cases(4.76%). The causes of death were: 3 cases of pulmonary hypertension crisis, 3 cases of severe mitral insufficiency(MI), 1 case of postoperative malignant arrhythmia, and the rest of the children were cured and discharged. Permanent pacemaker was installed in 3 patients due to atrioventricular block(AVB). The follow-up time was 1-10 years old, and 2 patients died in late stage: 1 patient did not seek medical treatment in time due to infection, and 1 patient had unknown cause. Five patients underwent secondary surgery: 4 due to severe mitral/tricuspid insufficiency(MI/TI) and 1 due to delayed AVB. The mid-term follow-up showed 9 cases of severe MI and 4 cases of severe TI. Compared with children with surgical age<3 months and ≥3 months, there were statistically significant differences in postoperative ventilator-assisted ventilation time, severe MI before postoperative discharge and total mortality between the two groups( P<0.05). Mid-term follow-up results showed no difference between the two groups. There were statistically significant differences in surgical age, postoperative CICU stay time and total hospital stay between the children with trisomy 21-syndrome and those without trisomy 21-syndrome( P<0.05), and there was no difference between the two groups in mid-term follow-up results. Residual shunt of 1-3 mm VSD was found in 29 cases, 26 cases were closed during follow-up, and 3 cases had smaller residual shunt. Conclusion:Modified single patch technique treatment of CAVSD has good effect, low mortality and low re-operation rate. But age <3 months group, infant mortality was significantly increased, the duration of postoperative mechanical assisted ventilation was prolonged, and the proportion of early postoperative severe MI was high.Severe MI and TI is easy to occur after CAVSD, which requires long-term follow-up and timely treatment. The children with trisomy 21-syndrome were similar to those with normal chromosome except for longer stay in ICU and total hospital stay.