Objective To improve the efficiency of induced differentiation of primitive neural epithelial cells derived from human induced pluripotent stem cells(hiPSCs-NECs)into functional midbrain dopaminergic progenitor cells(DAPs).Methods HiPSCs were cultured in mTeSRTM medium containing DMH1(10 μmol/L),SB431542(10 μmol/L),SHH(200 ng/mL),FGF8(100 ng/mL),purmorphamine(2 μmol/L),CHIR99021(3 μmol/L),and N2(1%)for 12 days to induce their differentiation into primitive neuroepithelial cells(NECs).The hiPSCs-NECs were digested with collagenase IV and then cultured in neurobasal medium supplemented with 1%N2,2%B27-A,BDNF(10 ng/mL),GDNF(10 ng/mL),AA,TGF-β,cAMP,and 1%GlutaMax in the presence of different concentrations of Rho kinase inhibitor Y27632,and the culture medium was changed the next day to remove Y27632.Continuous induction was performed until day 28 to obtain DAPs.Results Human iPSCs expressed the pluripotency markers OCT4,SOX2,Nanog,and SSEA1 and were positive for alkaline phosphatase staining.The hiPSCs-NECs were obtained on day 13 in the form of neural rosettes expressing neuroepithelial markers SOX2,nestin,and PAX6.In digested hiPSCs-NECs,the addition of 5 μmol/L Y27632 significantly promoted survival of the adherent cells,increased cell viability and the proportion of S-phase cells(P<0.01),and reduced the rate of apoptotic cells(P<0.05).On day 28 of induction,the obtained cells highly expressed the specific markers of DAPS(TH,FOXA2,NURR1,and Tuj1).Conclusion Treatment with Y27632(5 μmol/L)for 24 h significantly promotes the survival of human iPSCs-NECs during their differentiation into DPAs without affecting the cell differentiation,which indirectly enhances the efficiency of cell differentiation.

Objective To improve the efficiency of induced differentiation of primitive neural epithelial cells derived from human induced pluripotent stem cells(hiPSCs-NECs)into functional midbrain dopaminergic progenitor cells(DAPs).Methods HiPSCs were cultured in mTeSRTM medium containing DMH1(10 μmol/L),SB431542(10 μmol/L),SHH(200 ng/mL),FGF8(100 ng/mL),purmorphamine(2 μmol/L),CHIR99021(3 μmol/L),and N2(1%)for 12 days to induce their differentiation into primitive neuroepithelial cells(NECs).The hiPSCs-NECs were digested with collagenase IV and then cultured in neurobasal medium supplemented with 1%N2,2%B27-A,BDNF(10 ng/mL),GDNF(10 ng/mL),AA,TGF-β,cAMP,and 1%GlutaMax in the presence of different concentrations of Rho kinase inhibitor Y27632,and the culture medium was changed the next day to remove Y27632.Continuous induction was performed until day 28 to obtain DAPs.Results Human iPSCs expressed the pluripotency markers OCT4,SOX2,Nanog,and SSEA1 and were positive for alkaline phosphatase staining.The hiPSCs-NECs were obtained on day 13 in the form of neural rosettes expressing neuroepithelial markers SOX2,nestin,and PAX6.In digested hiPSCs-NECs,the addition of 5 μmol/L Y27632 significantly promoted survival of the adherent cells,increased cell viability and the proportion of S-phase cells(P<0.01),and reduced the rate of apoptotic cells(P<0.05).On day 28 of induction,the obtained cells highly expressed the specific markers of DAPS(TH,FOXA2,NURR1,and Tuj1).Conclusion Treatment with Y27632(5 μmol/L)for 24 h significantly promotes the survival of human iPSCs-NECs during their differentiation into DPAs without affecting the cell differentiation,which indirectly enhances the efficiency of cell differentiation.

Objective To compare the consistency of liquid chromatography-tandem mass spectrometry(LC-MS/MS)and fully auto-mated chemiluminescent immunoassay(CLIA)methods in measuring plasma aldosterone concentration(PAC)in the elderly patients in intensive care unit(ICU)and to explore the correlation between the levels of aldosterone(ALD)and blood biomarkers.Methods A total of 41 elderly ICU patients were included.PAC was measured using both LC-MS/MS and CLIA methods,followed by methodolo-gy validation and consistency comparison.Meanwhile,a retrospective analysis of the patients'clinical data was conducted,and the cor-relations between ALD and blood biomarkers were analyzed.Results The linear range of the LC-MS/MS method was 15 to 1 500 pg/mL,with a lower limit of quantification of 15 pg/mL.The inter-day and intra-day precisions were both less than 15％,and the re-covery rate was 94％to 99％,with no significant matrix effect.The PAC results detected by CLIA and LC-MS/MS methods were posi-tively correlated(r=0.762 7,P＜0.01),but the consistency between the two methods was poor.Deming regression analysis yielded the equation Y=0.969 6X-16.71,with a slope of 0.970(95％CI:0.890～1.049)and an intercept of-16.71(95％CI:-25.690 to-7.728).Bland-Altman analysis showed that CLIA overestimated PAC by an average of 21.18 pg/mL(95％CI:-25.89 to 68.26 pg/mL)compared to LC-MS/MS,with a bias of 24.02％.In the correlation analysis between ALD and blood biomarkers,ALD showed a significant positive correlation with myoglobin(Mb)(r=0.303,P＜0.05).Conclusion The LC-MS/MS method demonstrated good methodological performance in measuring PAC in the elderly patients in ICU.The consistency between LC-MS/MS and CLIA methods was poor,and the two methods should not be used interchangeably in clinical practice.There was a positive correlation between ALD and Mb,suggesting that ALD may be associated with myocardial injury.

Objective:To analyze the clinical characteristics of patients with Vibrio vulnificus infection, share diagnosis and treatment experience, and establish a rapid diagnosis procedure for this disease. Methods:This study was a retrospective case series study. From January 2009 to November 2022, 11 patients with Vibrio vulnificus infection who met the inclusion criteria were admitted to the Department of Burns and Wound Repair of Guangdong Provincial People's Hospital Affiliated to Southern Medical University. The gender, age, time of onset of illness, time of admission, time of diagnosis, route of infection, underlying diseases, affected limbs, clinical manifestations and signs on admission, white blood cell count, hemoglobin, platelet count, C-reactive protein (CRP), alanine transaminase (ALT), aspartate transaminase (AST), creatinine, procalcitonin, albumin, N-terminal pro-B-type natriuretic peptide (NT-proBNP), and blood sodium levels on admission, culture results and metagenomic next-generation sequencing (mNGS) results of pathogenic bacteria and the Vibrio vulnificus drug susceptibility test results during hospitalization, treatment methods, length of hospital stay, and outcomes of all patients were recorded. Comparative analysis was conducted on the admission time and diagnosis time of patients with and without a history of exposure to seawater/marine products, as well as the fatality ratio and amputation of limbs/digits ratio of patients with and without early adequate antibiotic treatment. For the survived patients with hand involvement, the hand function was assessed using Brunnstrom staging at the last follow-up. Based on patients' clinical characteristics and treatment conditions, a rapid diagnosis procedure for Vibrio vulnificus infection was established. Results:There were 7 males and 4 females among the patients, aged (56±17) years. Most of the patients developed symptoms in summer and autumn. The admission time was 3.00 (1.00, 4.00) d after the onset of illness, and the diagnosis time was 4.00 (2.00, 8.00) d after the onset of illness. There were 7 and 4 patients with and without a history of contact with seawater/marine products, respectively, and the admission time of these two types of patients was similar ( P>0.05). The diagnosis time of patients with a history of contact with seawater/marine products was 2.00 (2.00, 5.00) d after the onset of illness, which was significantly shorter than 9.00 (4.25, 13.00) d after the onset of illness for patients without a history of contact with seawater/marine products ( Z=-2.01, P<0.05). Totally 10 patients had underlying diseases. The affected limbs were right-hand in 8 cases, left-hand in 1 case, and lower limb in 2 cases. On admission, a total of 9 patients had fever; 11 patients had pain at the infected site, and redness and swelling of the affected limb, and 9 patients each had ecchymosis/necrosis and blisters/blood blisters; 6 patients suffered from shock, and 2 patients developed multiple organ dysfunction syndrome. On admission, there were 8 patients with abnormal white blood cell count, hemoglobin, and albumin levels, 10 patients with abnormal CRP, procalcitonin, and NT-proBNP levels, 5 patients with abnormal creatinine and blood sodium levels, and fewer patients with abnormal platelet count, ALT, and AST levels. During hospitalization, 4 of the 11 wound tissue/exudation samples had positive pathogenic bacterial culture results, and the result reporting time was 5.00 (5.00, 5.00) d; 4 of the 9 blood specimens had positive pathogenic bacterial culture results, and the result reporting time was 3.50 (1.25, 5.00) d; the mNGS results of 7 wound tissue/exudation or blood samples were all positive, and the result reporting time was 1.00 (1.00, 2.00) d. The three strains of Vibrio vulnificus detected were sensitive to 10 commonly used clinical antibiotics, including ciprofloxacin, levofloxacin, and amikacin, etc. A total of 10 patients received surgical treatment, 4 of whom had amputation of limbs/digits; all patients received anti-infection treatment. The length of hospital stay of 11 patients was (26±11) d, of whom 9 patients were cured and 2 patients died. Compared with that of the 6 patients who did not receive early adequate antibiotic treatment, the 5 patients who received early adequate antibiotic treatment had no significant changes in the fatality ratio or amputation of limbs/digits ratio ( P>0.05). In 3 months to 2 years after surgery, the hand function of 8 patients was assessed, with results showing 4 cases of disabled hands, 2 cases of incompletely disabled hands, and 2 cases of recovered hands. When a patient had clinical symptoms of limb redness and swelling and a history of contact with seawater/marine products or a pre-examination triage RiCH score of Vibrio vulnificus sepsis ≥1, the etiological testing should be initiated immediately to quickly diagnose Vibrio vulnificus infection. Conclusions:Vibrio vulnificus infection occurs most frequently in summer and autumn, with clinical manifestations and laboratory test results showing obvious infection characteristics, and may be accompanied by damage to multiple organ functions. Both the fatality and disability ratios are high and have a great impact on the function of the affected limbs. Early diagnosis is difficult and treatment is easily delayed, but mNGS could facilitate rapid detection. For patients with red and swollen limbs accompanied by a history of contact with seawater/marine products or with a pre-examination triage RiCH score of Vibrio vulnificus sepsis ≥1, the etiological testing should be initiated immediately to quickly diagnose Vibrio vulnificus infection.

OBJECTIVE To compare the safety of high-dose methotrexate （HD-MTX） via peripherally inserted central catheter （PICC） and totally implantable venous access port （TIVAP） in pediatric patients with malignant brain tumors. METHODS Patients with malignant brain tumors who received HD-MTX via PICCs or TIVAPs in our hospital from July 2018 to April 2022 were retrospectively analyzed. Clinical data were collected to compare differences in blood concentration of methotrexate （MTX），the incidence of adverse events （including adverse drug reactions and catheter-related complications） and length of stay in hospital. Multivariate linear regression was applied to analyze the factors that influenced the blood concentration of MTX. RESULTS A total of 107 patients were included in the study，with 65 patients in the PICC group and 42 patients in the TIVAP group. Blood concentration of MTX at 24 h （C24 h） in TIVAP group was significantly higher than PICC group （[ 126.87±61.99） μmol/L vs. （102.45±48.77） μmol/L，P＜0.05）. There was no significant difference in blood concentration of MTX at 42 h （C42 h），compared with PICC group （P＞0.05）. Results of multivariate linear regression analysis showed that TIVAP was associated with the increase of C24 h（P＜0.05）. No significant differences were observed in the incidence of adverse events and the length of stay in the hospital between 2 groups （P＞0.05）. CONCLUSIONS Risk of adverse events is not increased，although the MTX C24 h level is elevated after administration of TIVAP. TIVAP is a safe choice for HD-MTX therapy with implementing therapeutic drug monitoring.

Objective:To explore the obstacle factors and promoting factors of exercise guidance and implementation after breast cancer surgery, so as to provide reference and basis for the formulation of exercise program after breast cancer patients.Methods:Using the objective sampling method, a total of 10 medical staff and 10 patients with breast cancer after operation from the breast surgery department of the Seven People's Hospital Affiliated to Shanghai University of Traditional Chinese Medicine hospital in Shanghai were selected as research objects from July to September 2022. A semi-structured interview was conducted. The interview outline was developed based on the knowledge transformation framework, and the results were analyzed by the content analysis method.Results:The study extracted three levels of influencing factors. The system level included caregiver workload, educational materials, norms and responsibilities and trust in evidence-based evidence. The practitioner level included the level of exercise knowledge, the concern about patient compliance and the emphasis on exercise. The patient level included sports knowledge level, personality and interest, venous line, adverse reaction of treatment, psychological state, weather environment, attention to health and concern of family members.Conclusions:The medical staff need to consider the postoperative exercise guidance for breast cancer patients from the department, themselves and patients, and should fully consider the patients' conditions and subjective experience to improve the patients' exercise compliance.

Objective:To evaluate the value of 18F-FDG PET/CT in the diagnosis and treatment of primary breast lymphoma (PBL). Methods:Clinical data and 18F-FDG PET/CT imaging data of 6 patients (all females, age 46-79 years) with pathologically diagnosed primary breast diffuse large B cell lymphoma (PB-DLBCL) in Xishan People′s Hospital of Wuxi City and Affiliated Hospital of Jiangnan University from July 2015 to October 2021 were analyzed retrospectively. A total of 10 18F-FDG PET/CT scans were done for primary staging (6 scans of 6 patients), evaluation of treatment response (3 scans of 2 patients), and recurrence detection (1 scan of 1 patient). 18F-FDG PET/CT image analysis was performed qualitatively (visually) and semi-quantitatively (SUV max). Treatment response was evaluated by Deauville scores. Results:All 6 patients were diagnosed pathologically as PB-DLBCL (3 patients by core needle biopsy, 3 patients by biopsy after lumpectomy). All 6 patients were staged using baseline 18F-FDG PET/CT before chemotherapy. For 3 patients diagnosed by core needle biopsy, baseline 18F-FDG PET/CT showed unilateral breast lesion with high FDG uptake (SUV max: 23.0, 52.9, and 33.6). For 3 postoperative patients, baseline 18F-FDG PET/CT showed flocculent soft tissue density in the operative area with low FDG uptake (SUV max: 3.4, 2.2 and 2.0). Patient No.2 showed a large left breast mass with left axillary lymph node involvement by baseline PET/CT, and multiple nodular uptakes in bilateral breast (Deauville score of 4) after 4 courses of chemotherapy and negative result (Deauville score of 1) after 3 courses of new chemotherapy regimens by PET/CT. Patient No.4 showed right breast lesion and right axillary lymph nodes by routine preoperative imaging examination, but left breast lesion by postoperative PET/CT. According to the results of 18F-FDG PET/CT, patient No.4 was with complete response (Deauville score of 1) after treatment, but recurrence (Deauville score of 5) occurred after 7 months follow-up. Conclusion:18F-FDG PET/CT can play an important role in every step of management (diagnosis and staging, treatment response evaluation and detection of recurrence) in patients with PB-DLBCL.

Objective:To investigate the safety and the efficacy of percutaneous and surgical approach in femoro-femoral veno-arterial extracorporeal membrane oxygenation (VA-ECMO) cannulation.Methods:All consecutive patients implanted with femoro-femoral VA-ECMO between January 2018 and December 2020 in Beijing Anzhen Hospital, Capital Medical University. Propensity score matching was used to compare outcomes of percutaneous and surgical groups while controlling for confounders.Results:Among the 276 patients who received femoro-femoral VA-ECMO (62 surgical and 214 percutaneous), propensity-score matching selected 52 pairs of patients with similar characteristics with mean age of(59.6±13.0)years old, in which 26 patients were female. There were a lower ECMO cannulation-associated complication (28.8% vs. 48.1%, P=0.044) and a lower hospital mortality (42.3% vs. 67.3%, P=0.010) in the percutaneous group. The circuit blood flow after ECMO initiation was similar in both groups[(3.3±0.8)L·min -1·kg -1 in percutaneous group vs. (3.2±0.7)L·min -1·kg -1 in surgical group, P=0.738]. The serum lactate was declined in both group after ECMO initiation[(5.4±5.8)mmol/L vs. (9.2±6.9)mmol/L, P<0.001 in percutaneous group; (6.3±6.2)mmol/L vs. (10.5±7.0)mmol/L, P=0.003 in surgical group]. Conclusion:Percutaneous approach is a safe and efficient technique in emoro-femoral VA-ECMO cannulation. Compared with surgical cannulation, percutaneous approach is associated with lower ECMO cannulation-associated complication and lower hospital mortality.

To investigate the effect of dietary fiber on blood glucose and pregnancy outcomes in patients with gestational diabetes mellitus (GDM). One hundred and twelve patients with GDM in the second trimester of pregnancy were recruited from Women's Hospital, Zhejiang University School of Medicine. Patients were randomized into two groups with 56 in each group: the control group received basic nutrition support; while the dietary fiber group were given additional dietary fiber ( total dietary fiber per day) before meals in addition to basic nutrition support. Intervention for all cases lasted for 8 weeks. Fasting blood glucose and postprandial blood glucose (2 h BG) were measured every week, and oral glucose tolerance test (OGTT) was performed at 42 d postpartum to evaluate the glycemic outcomes. Perinatal outcomes were recorded. The dietary fiber intervention markedly improved 2 h BG in patients with GDM and significantly elevated the glucose compliance rate from the 3rd to 8th week compared to the control group ( <0.05 or <0.01). OGTT 2 h glucose and the incidence of impaired glucose tolerance in the dietary fiber group were significantly lower than those in the control group, while the glucose compliance rate was significantly higher than that in the control group (all <0.01). Moreover, the rates of adverse perinatal outcomes, such as premature rupture of membranes and neonatal hyperbilirubinemia were declined in the dietary fiber group (<0.05 or <0.01). Dietary fiber intervention can ameliorate hyperglycemia in GDM patients, improve perinatal outcomes and reduce the incidence of postpartum impaired glucose tolerance.
Blood Glucose ; Diabetes, Gestational ; Dietary Fiber ; Female ; Glucose Tolerance Test ; Humans ; Infant, Newborn ; Pregnancy ; Pregnancy Outcome

【Objective】 To investigate the clinical staffs′ knowledge about blood transfusion and their demands, so as to provide evidence for arranging the content for continuing education of blood transfusion medicine. 【Methods】 A self-designed questionnaire was used to investigate the blood transfusion related knowledge of 652 clinical staffs from other cities or counties/districts in Shaanxi Province, who came to our hospital to attend meetings, further training and study from July to August 2019. 【Results】 Clinical stsffs investigated lacked a deep understanding of blood transfusion and blood donation related knowledge, and even were ignorant of certain knowledge. The awareness rate of relevant knowledge was the highest among doctors, followed by laboratory technicians and the lowest among nurses. 【Conclusion】 The awareness rate of blood transfusion related knowledge among medical staffs is relatively low, suggesting the training of blood transfusion related knowledge should be enhanced.