Objective To investigate the clinical efficacy of low-dose Prednisone combined with Tacrolimus in children with refractory nephrotic syndrome(RNS).Methods A total of 106 children with RNS admitted from March 2022 to September 2023 were included and evenly divided into the monotherapy group and the combination therapy group according to the random number table method.The monotherapy group was given low-dose Prednisone tablets,while the combination therapy group was given low-dose Prednisone tablets combined with Tacrolimus.The differences in nailfold microcirculation indicators,lipid metabolism indicators,renal injury markers,clinical efficacy,and adverse reactions were compared between the two groups.Results The total effective rate of the combination therapy group was higher than that of the monotherapy group(P<0.05).At 3 months after treatment,the serum albumin,capillary flow velocity,input branch diameter and output branch diameter in the combination therapy group were all higher than those in the monotherapy group,while the levels of red blood cell aggregation,triglycerides,low-density lipoprotein cholesterol,total cholesterol,serum creatinine,24-hour urine protein quantification,neutrophil gelatin enzyme-associated lipocalin,retinol binding protein,α1-microglobulin,β-N-acetylglucosaminidase,and renal injury molecule-1 were all lower than those in the monotherapy group(P<0.05).The total incidence of adverse reactions in the combination therapy group was lower than that in the monotherapy group(P<0.05).Conclusion In the treatment of children with RNS,the combination of Tacrolimus and low-dose Prednisone can improve the microcirculation status and reduce renal injury markers,with remarkable effects.

OBJECTIVE To provide a reference for optimizing and promoting the quality standards of Shechuan naolitong granules. METHODS Fifteen batches of Shechuan naolitong granules were used as samples to establish HPLC fingerprints using the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine （2012 edition）. Similarity evaluation and common peak identification were performed， and orthogonal partial least squares discriminant analysis （OPLS-DA） was used to assess quality differences among different batches and to screen quality differential components. Using salvianolic acid B（SAB） as the internal reference， quantitative analysis of multi-components by single-marker （QAMS） was developed to simultaneously determine geniposidic acid （GA）， chlorogenic acid （CA）， vaccarin （VA）， ferulic acid （FA） and senkyunolide I （SI）. The results were compared with those obtained by the external standard method. RESULTS A total of 13 common peaks were identified in the HPLC fingerprints of 15 batches of samples， and the similarities of the spectra were all above 0.96. Seven chromatographic peaks were identified as GA （peak 3）， CA （peak 6）， VA （peak 8）， FA （peak 9）， SI （peak 11）， SAB（peak 12） and TA（peak 13）. OPLS-DA indicated that the differential quality markers among 15 batches were peaks 5， 11 （SI）， and 12 （SAB）.Using SAB as the internal reference， the relative correction factors for GA， CA， VA， FA and SI were calculated as 1.058 4， 0.594 3， 0.643 3， 0.342 7 and 0.262 8， respectively. The mean content of GA， CA， VA， FA， SI and SAB across the 15 batches of samples were 0.155 0， 0.085 4， 0.140 3， 0.071 8， 0.072 7， 1.276 3 mg/g， respectively， showing no significant difference compared with the ESM （P＞0.05）. CONCLUSIONS The established HPLC fingerprint and QAMS are simple， efficient and economical， providing a reference for the quality control and further development of Shechuan naolitong granules.

Objective To investigate the clinical efficacy of low-dose Prednisone combined with Tacrolimus in children with refractory nephrotic syndrome(RNS).Methods A total of 106 children with RNS admitted from March 2022 to September 2023 were included and evenly divided into the monotherapy group and the combination therapy group according to the random number table method.The monotherapy group was given low-dose Prednisone tablets,while the combination therapy group was given low-dose Prednisone tablets combined with Tacrolimus.The differences in nailfold microcirculation indicators,lipid metabolism indicators,renal injury markers,clinical efficacy,and adverse reactions were compared between the two groups.Results The total effective rate of the combination therapy group was higher than that of the monotherapy group(P<0.05).At 3 months after treatment,the serum albumin,capillary flow velocity,input branch diameter and output branch diameter in the combination therapy group were all higher than those in the monotherapy group,while the levels of red blood cell aggregation,triglycerides,low-density lipoprotein cholesterol,total cholesterol,serum creatinine,24-hour urine protein quantification,neutrophil gelatin enzyme-associated lipocalin,retinol binding protein,α1-microglobulin,β-N-acetylglucosaminidase,and renal injury molecule-1 were all lower than those in the monotherapy group(P<0.05).The total incidence of adverse reactions in the combination therapy group was lower than that in the monotherapy group(P<0.05).Conclusion In the treatment of children with RNS,the combination of Tacrolimus and low-dose Prednisone can improve the microcirculation status and reduce renal injury markers,with remarkable effects.

Objective In this study,we compared the masticatory efficiency,occlusal functional indexes,and satisfaction of lingual-ized occlusion(LO)vs.anatomic occlusion(AO)and discussed the difference between these occlusions.Methods Twenty edentulous patients were included in this clinical trial during 2021 April to December.They were divided into 2 groups according to their denture occlusion.Masticatory efficiency was tested by chewing the same amount of peanuts(by weight)and then the absorption of the solution was compared under a spectrophotometer after wearing dentures for 1 month.The TeeTester occlusion analysis system was used to test masticatory function,and satisfaction research was conducted for the edentulous patients at this time point as well.Finally,statistical analyses were conducted to compare the difference in masticatory efficiency between AO and LO by SPSS 20.0.Results The indexes of LO were worse than AO in masticatory efficiency,masticatory function and most of satisfaction research,while LO was better than AO in speech recovery.The difference was only statistically significant in COF-y of masticatory function(P＜0.05).Conclusion LO is similar with AO in terms of masticatory efficiency,satisfaction and denture stability;LO has good performance in denture's fit and use for patients.

CRISPR-Associated Protein 9 ; Gene Editing ; CRISPR-Cas Systems/genetics*

Objective:To analyze the clinical characteristics of elderly acute pulmonary thromboembolism(APE)patients complicated with preexisting atrial fibrillation(AF)and the impact of preexisting AF on in-hospital adverse outcomes in elderly patients with APE.Methods:A retrospective analysis was performed on elderly APE patients with preexisting AF hospitalized in Beijing Anzhen Hospital, Capital Medical University between January 1, 2008 and December 31, 2021.We compared the comorbidities, symptoms, signs, laboratory test results and echocardiographic features, simplified pulmonary embolism severity index(sPESI)scores and adverse in-hospital outcomes between the preexisting AF group and the non-AF group.Logistic regression was used to analyze the risk factors of in-hospital adverse outcomes in elderly patients with APE.Results:A total of 240 patients diagnosed with APE were enrolled.There were 120 patients in the AF group and 120 patients in the non-AF group.For patients in the AF group and the non-AF group, the proportions with chronic heart failure were 38.3%(46/120)and 15.8%(19/120), the proportions with lower extremity deep vein thrombosis(DVT)were 36.7%(44/120)and 65.8%(79/120), the left ventricular ejection fractions(LVEF)were(59±10)% and(62±7)%, and hospital stays were(15±7)and(11±4)days, respectively, and the differences were statistically significant( χ2=15.381, 20.429, t=2.527, -4.710, all P<0.05). The incidences of in-hospital adverse outcomes in the AF group and the non-AF group were 4.2%(5/120)and 3.3%(4/120), respectively, with no significant difference( χ2=0.000, P=1.000). The overall incidence of in-hospital adverse outcomes was 3.8%(9/240). Multivariate Logistic regression analysis showed that elevated lactic acid was an independent risk factor for in-hospital adverse outcomes( OR=2.753, 95% CI: 1.367-5.542, P=0.005). However, AF( OR=2.880, 95% CI: 0.587-14.141, P=0.192)and sPESI score( OR=2.056, 95% CI: 0.904-4.673, P=0.086)were not associated with in-hospital adverse outcomes. Conclusions:Elderly APE patients with preexisting AF have a relatively low incidence of DVT, but a higher proportion have concurrent chronic heart failure and need a longer hospital stay.Elevated lactic acid is an independent risk factor for in-hospital adverse outcomes of elderly APE patients with preexisting AF.However, preexisting AF has no predictive value for in-hospital adverse outcomes in elderly patients with APE.

Objective:To explore the application effect of adjustable body position protection belt in patients undergoing not painless colonoscopy.Methods:A randomized and controlled trial was used, a total of 180 patients who planned to undergo not painless colonoscopy in the Endoscopy center, the First Hospital of Jiaxing from January to October 2022 were selected as the research objects. They were divided into the experimental group and the control group by the random number table method, with 90 cases in each group. The control group adopted the traditional method of colonoscopy body position placement, and on this basis, the experimental group used the adjustable body position protection belt to assist the patient body position placement. The maintenance rate of body position in colonoscopy, the cecal intubation time, the degree of intraoperative pain and the acceptance rate of re-examinationunder the same operating doctor and nurse were compared between the two groups.Results:The maintenance rate of body position in colonoscopy was 97.78%(88/90) in the experimental group and 58.89%(53/90) in the control group, the difference was statistically significant ( χ2 = 37.84, P<0.05). The cecal intubation time was (7.84 ± 4.39) min in the experimental group and (10.13 ± 5.85) min in the control group, the difference was statistically significant ( t = 2.98, P<0.05). The intraoperative pain score was 2.96 ± 1.39 in the experimental group and 4.28 ± 1.42 in the control group, the difference was statistically significant ( t = 6.31, P<0.05). The acceptance rate for re-colonoscopy was 88.89%(80/90) in the experimental group and 72.22%(65/90) in the control group, the difference was statistically significant ( χ2 = 7.98, P<0.05). Conclusions:Use of adjustable body position protection belt to assist body position placement before the patients undergoing not painless colonoscopy, it can improve the maintenance rate of body position during not painless colonoscopy, shorten the time of cecal intubation, reduce the intraoperative pain degree, and patients also have a higher willingness to undergo colonoscopy again.

Objective:To evaluate the efficacy of allogeneic hematopoietic cell transplantation(allo-HSCT) using unrelated cord blood or haploidentical donors in the treatment of children with primary immunodeficiency diseases (PID).Methods:The clinical data of 60 children with PID admitted to Chinese People′s Liberation Army General Hospital-Sixth Medical Center from April 2014 to October 2019 were retrospectively analyzed, including 56 cases of chronic granulomatous disease, 2 cases of severe combined immunodeficiency disease, 1 case of high-IgM syndrome and 1 case of severe congenital neutropenia.All patients underwent allo-HSCT, including 12 cases receiving the transplantation from unrelated cord blood (UCB group) and 48 cases from haploidentical donors combined with a third party unrelated cord blood (haploid group). Among these patients, there were 59 males and 1 female, with a median age of 3.4 years.All patients received a myeloablative conditioning regimen based on Busulfan.The prophylaxis of acute graft versus host disease (aGVHD) was performed based on Cyclosporine.In the UCB group, the median dose of mononuclear cells and CD 34+ cells was 0.67×10 8/kg and 0.51×10 6/kg recipient body weight, respectively; In the haploid group, bone marrow and peripheral stem cells from haploid donors were infused on day 01 and day 02, respectively.The third party cord blood was infused 4 hours before bone marrow infusion.The median dose of mononuclear cells and CD 34+ cells of bone marrow and peripheral stem cells from haploid donors was 9.97×10 8/kg and 5.12×10 6/kg recipient body weight, respectively.Kaplan-Meier method was used to analyze the overall survival rate. Results:The median day to neutrophil and platelet engraftment was 13.0 days and 23.5 days, respectively.The rate of complete donor chime-rism was shown 30.0 days after transplantation.There was no case with primary engraftment failure, and 1 case with secondary engraftment failure.The incidence of grade Ⅰ-Ⅱ and grade Ⅲ-Ⅳ aGVHD was 43.3% and 15.5%, respectively.The incidence of chronic graft versus host disease with limited skin type was 6.7%, while that with extensive type was 1.1%.The median follow-up period was 818 days.There were 6 death cases, among which, 5 cases died from infection and 1 case died from heart failure.The total mortality related to transplantation was 11.9%.A total of 53 cases survived without diseases.The estimated 5-year failure free survival and overall survival rate was 83.9% and 88.1%, respectively.Conclusion:The efficacy of allo-HSCT in the treatment of children with PID using unrelated cord blood and haploidentical donors is favorable.

Objective     To evaluate the early clinical effect of reimplantation in the treatment of bicuspid aortic valve (BAV) with aortic root aneurysm. Methods     The clinical data of 25 patients with BAV and aortic root aneurysm [mean diameter: 45-63 (52.68±5.55) mm] undergoing reimplantation in West China Hospital from November 2019 to May 2021 were retrospectively reviewed. There were 22 males and 3 females. The mean age was 15-65 (50.00±13.10) years and body surface area was 1.79±0.23 m2. Results    The pathological classification of BAV malformation was confirmed during the operation: Type 0 in 3 patients and Type 1 in 22 patients. There were 12 patients undergoing cusp central plication, and 2 patients were sutured with a closed fusion crest. Postoperative valve leaflet coaptation height was 0.78±0.15 cm, and effective height was 1.27±0.19 cm. In operation, maximum aortic valve flow velocity was 1.65±0.42 m/s, pressure difference was 5.46±3.05 mm Hg, and aortic valve annulus diameter was 21.32±0.95 mm. Cardiopulmonary bypass time was 225.84±35.34 min, and aortic block time was 189.60±26.51 min. In-hospital time was 11.64±3.07 d, ICU stay time was 2.64±0.99 d, and mechanical ventilation time was 1.48±0.87 d. The follow-up time was 17.20±4.70 months, and no death or major complications occurred during the follow-up in all patients. The cardiac function of the patients significantly improved postoperatively (P≤0.05). Echocardiography suggested that 12 patients had no aortic regurgitation, 10 minor aortic regurgitation, 3 mild aortic regurgitation, and no patients with moderate or more severe regurgitation. The diameter of the aortic sinus, left ventricular end-diastolic diameter and volume decreased during the follow-up, compared to preoperative ones (P≤0.05). The maximum flow velocity of the aortic valve was 1.54±0.36 m/s, and the pressure difference was 5.17±2.38 mm Hg during the follow-up. Conclusion    Reimplantation technology has a good clinical effect for highly selective BAV patients. It can effectively avoid long-term postoperative anticoagulation, but the maximum flow rate after surgery is slightly increased, which may be related to the configuration of BAV itself. While compared with valve replacement, the effect is still worthy of recognition.

Objective:To examine the effects of apelin-13 on ferroptosis of the C2C12 skeletal muscle cell line induced by a high-iron environment and explore potential underlying mechanisms.Methods:C2C12 cells were cultured in Dulbecco's Modified Eagle Medium(DMEM)and divided into a control group, a ferric citrate(FAC)group, an apelin-13 group, an FAC+ apelin-13 group, a ferroptosis inducer RSL3 group and an FAC+ apelin-13+ RSL3 group.Cell viability was detected by the 3-(4, 5-dimethyl thiazole-2)-2, 5-diphenyl thiazolyl blue(MTT)assay.The intracellular concentrations of total iron and divalent iron were measured by colorimetry; the levels of glutathione(GSH), malondialdehyde(MDA)and intracellular reactive oxygen species(ROS)in cells were detected by an enzyme-linked immunosorbent assay, visible spectrophotometry and a chemifluorescence method, respectively.The ultrastructure of C2C12 cells was examined by transmission electron microscopy.The protein expression of glutathione peroxidase 4(GPX-4), ferritin heavy chain 1(FTH-1), heme oxygenase 1(HO-1)and nuclear factor E2-related factor 2(Nrf-2), were detected by Western blotting.Results:Compared with the FAC group, the FAC+ Apelin-13 group had significantly elevated cell viability(optical density: 0.52±0.06 vs.0.28±0.04, t=7.837, P=0.007)and higher concentrations of GSH(2.41±0.35 vs.0.91±0.12 μmol/g Pro, t=9.778, P=0.003), but significantly decreased levels of ROS(22.06±5.79 vs.52.71±7.28 a. u./mg Pro, t=8.064, P=0.006), MDA(4.63±0.51 vs.9.11±0.84 mmol/mg Pro, t=8.642, P=0.006), total iron(1.53±0.24 vs.3.17±0.55 μmol/g Pro, t=6.135, P=0.013)and divalent iron(0.75±0.08 vs.1.94±0.36 μmol/g Pro, t=5.068, P=0.027), as well as reduced intracellular iron deposition.In the control group and the apelin-13 group, the morphology of the mitochondria was clear and normal.In contrast, the mitochondria in the FAC group had increased membrane density, membrane shrinkage and rupture, vacuolar degeneration, and obvious mitochondrial damage, which were consistent with the morphological characteristics of ferroptosis.Compared with the FAC group, the FAC+ apelin-13 group showed significant improvement in mitochondrial damage.Moreover, compared with the FAC+ apelin-13 group, the cell viability of the FAC+ apelin-13+ RSL3 group was significantly decreased(optical density: 0.23±0.04 vs.0.48±0.06, t=7.642, P=0.007). Compared with the FAC group, the FAC+ apelin-13 group had significantly up-regulated cellular expression of GPX-4(relative expression: 0.96±0.14 vs.0.31±0.07, t=7.712, P=0.008), FTH-1(0.57±0.08 vs.0.27±0.05, t=6.944, P=0.011), and HO-1(0.49±0.07 vs.0.28±0.05, t=6.472, P=0.012), as well as increased nuclear expression of Nrf-2(relative expression: 0.42±0.04 vs.0.19±0.05, t=7.114, P=0.008)with a higher ratio of nuclear expression over total cellular expression[(58.36±5.24)% vs.(36.58±5.32)%, t=5.858, P=0.015]and a higher level of HO-1 protein expression(relative expression: 0.49±0.07 vs.0.28±0.05, t=6.472, P=0.012). Conclusions:Apelin-13 inhibits ferroptosis induced by a high iron environment in C2C12 cells, and the underlying molecular mechanisms may be related to the Nrf-2/HO-1 signaling pathway.