In vitro erythrocyte manufacturing technology offers a novel solution for transfusion therapy, ensuring sustainable blood supply, reducing dependence on donors, and minimizing the risk of infectious disease transmission. This review summarizes recent advancements in in vitro erythrocyte production, including techniques for generating red blood cells (RBC) from various cell sources, with a focus on the cultivation and development of immortalized erythroid cell lines. The importance of enhancing enucleation efficiency and ensuring the safety of immortalized erythroid cells is emphasized. Additionally, the potential and challenges in clinical translation are discussed, aiming to provide new insights for future sustainable RBC supply and therapeutic applications.

Objective:To establish a nomograph prediction model of early critical changes in children with febrile convulsion, and to provide guidance for the prevention and nursing of children with febrile convulsion.Methods:Convenient sampling method was adopted to select 384 children with febrile convulsion in Anhui Children ′s Hospital from January 2018 to April 2021 as the research objects. Based on pews, the children with febrile convulsion were divided into 334 cases of non risk group and 50 cases of risk group. Binary Logistic regression analysis were used to determine the independent risk factors affecting the early critical changes of children with febrile convulsion. A nomogram was drawn based on the independent risk factors. The discrimination and consistency of the model were verified by model ROC curve and Hosmer Lemeshow goodness of fit.The external validation of model prediction efficiency were verified by validation data. Results:Binary Logistic regression analysis showed that age, respiratory rhythm disorder, unconsciousness, breath rate, heart rate, neutrophil-to-lymphocyte ratio (NLR), red blood cell distribution width (RDW), duration of first convulsion and mean body temperature after first convulsion were influence factors for early critical changes in children with febrile convulsion ( P<0.05). The C-index of the model was 0.974 (95% CI 0.954-0.993), and the C-index of the external validation of the model was 0.922 (95% CI 0.880-0.966). The results of H-L fitting test showed that the difference was not statistically significant( χ2=0.29, P>0.05). Conclusions:The early critical changes of children with febrile convulsion may be affected by respiratory rhythm disorder, confusion of consciousness, breath rate, heart rate, NLR, RDW, duration of the first convulsion, mean temperature after the first convulsion and other factors. Pediatric emergency department should collect corresponding intervention measures for children with febrile convulsion according to the establishment of prediction model to prevent their early deterioration.

AIM: To investigate the relationship between serum soluble growth stimulation expressed gene 2 protein(sST2) level and coronary artery complex lesions and their severity. METHODS: A total of 430 patients, who were sequentially admitted to hospital for selective coronary artery angiography, were divided into control group (non-coronary heart disease group, 136 patients), simple lesions group of coronary heart disease (86 patients), complex lesions group (208 patients). To quantitative evaluate the complexity of coronary artery lesions, Syntax scores were further performed on patients in complex lesions groups, including 139 patients in the low-risk group, 36 patients in the medium-risk group, and 33 patients in the high-risk group. The serum soluble ST2 level of each group of patients was tested by means of ELISA. Spearman correlation analysis was used for the correlation between the level of soluble ST2 and the severity of coronary complex lesions. RESULTS: In 430 subjects, the soluble ST2 level of all patients with coronary heart disease (including simple lesions and complex lesions) was significantly higher than that of the control group [(3 449±1 250) vs. (2 743±961) pg/mL, P<0.001]; the sST2 levels of patients in the coronary artery simple lesions group, complex lesions low-risk group, medium-risk group and high-risk group were (3 200±1 406), (3 338±1 064), (3 728±1 228) and (4 261±1 235) pg/mL respectively, and the differences of sST2 levels among above four groups were statistically significant (P<0.001). Logistic regression analysis showed that sST2 was independently associated with coronary heart disease (OR=1.001, P<0.001) and sST2 was independently associated with the severity of coronary artery complex lesions (OR=1.001, P<0.001). Spearman-related analysis shows that the expression levels of sST2 are positively related to the severity of coronary artery lesions (rs: 0.543, P<0.001). The ROC curve showed that the area under the curve of sST2 for complex coronary lesions was AUC=0.726. CONCLUSION: Serum soluble ST2 level may be an important predictor of complicated coronary artery disease.

During the traumatic brain injury (TBI), improved expression of circulatory miR-21 serves as a diagnostic feature. Low levels of exosome-miR-21 in the brain can effectively improve neuroinflammation and blood-brain barrier (BBB) permeability, reduce nerve apoptosis, restore neural function and ameliorate TBI. We evaluated the role of macrophage derived exosomes-miR-21 (M-Exos-miR-21) in disrupting BBB, deteriorating TBI, and Rg1 interventions. IL-1

There is an imbalance between the supply and demand of functional red blood cells (RBCs) in clinical appli-cations. This imbalance can be addressed by regenerating RBCs using several in vitro methods. Induced pluripotent stem cells (iPSCs) can handle the low supply of cord blood and the ethical issues in embryonic stem cell research, and provide a promising strategy to eliminate immune rejection. However, no complete single-cell level differentiation pathway exists for the iPSC-derived erythroid differentiation system. In this study, we used iPSC line BC1 to establish a RBC regeneration system. The 10X Genomics single-cell transcriptome platform was used to map the cell lineage and differentiation trajectory on day 14 of the regeneration system. We observed that iPSC differentiation was not synchronized during embryoid body (EB) culture. The cells (on day 14) mainly consisted of mesodermal and various blood cells, similar to the yolk sac hematopoiesis. We identified six cell classifications and characterized the regulatory transcription factor (TF) networks and cell–cell contacts underlying the system. iPSCs undergo two transformations during the differentiation trajectory, accompanied by the dynamic expression of cell adhesion molecules and estrogen-responsive genes. We iden-tified erythroid cells at different stages, such as burst-forming unit erythroid (BFU-E) and orthochromatic erythroblast (ortho-E) cells, and found that the regulation of TFs (e.g., TFDP1 and FOXO3) is erythroid-stage specific. Immune erythroid cells were identified in our system. This study provides systematic theoretical guidance for optimizing the iPSC-derived erythroid differentiation system, and this system is a useful model for simulating in vivo hematopoietic develo-pment and differentiation.

Objective To report a case of Costello syndrome complicated by curis laxa,and to make a molecular genetic diagnosis.Methods Clinical data were collected from a case of Costello syndrome complicated by cutis laxa.Skin tissues were resected from the patient,and peripheral blood samples were obtained from the patient's parents and 150 unrelated healthy controls.Genomic DNA was extracted from these samples,and all the exons and their flanking sequences of the HRAS gene were analyzed by DNA sequencing.Results The 13-month-old female patient presented with growth retardation,severe malnutrition,coarse facial appearance,severely loose skin over the limbs,and decrease or disappearance of subcutaneous fat.A heterozygous mutation c.34G ＞ T (p.Gly12Cys) was detected in exon 2 of the HRAS gene in the patient,but not in her parents or 150 unrelated healthy controls.Conclusion The c.34G ＞ T (p.Gly12Cys) mutation in exon 2 of the HRAS gene may be responsible for Costello syndrome in the patient.

Objective To investigate the expression of STAT3 signaling pathway genes including Survivin and COX-2 in cholangiocarcinoma,as well as the relationship between expression of these genes and prognosis of patients with cholangiocarcinoma.Methods The tumor and normal tissue samples were respectively collected from 43 patients with cholangiocarcinoma and 12 patients with intra-and extrahepatic bile duct stones or hepatic duct injury in the Affiliated Provincial Hospital of Anhui Medical University from September 2007 to July 2012.The expression of STAT3,phosphorylated-STAT3 (p-STAT3),Survivin and COX-2 were examined using immunohistochemistry,and the relationship between the expression of these genes and the clinical pathological features and prognosis of patients with cholangiocarcinoma was analyzed.Patients were followed-up through outpatient examination and telephone interview until March 2014.Categorical data were analyzed using the chi-square test.Correlation analysis was done by Spearman's method.The survival curve was generated using the Kaplan-Meier method,and the survival analysis was conducted using the log-rank test.Results The positive expression rates of STAT3,p-STAT3,Survivin and COX-2 in the tumor samples were 69.8％ (30/43),65.1％ (28/43),72.1％ (31/43),79.1％ (34/43),respectively,which were compared with 41.7％ (5/12),8.3％ (1/12),16.7％ (2/12) and 41.7％ (5/12) in the normal tissue samples,showing a significant difference for the last 3 indexes (x2=12.136,9.811,4.679,P ＜ 0.05).Overexpression of p-STAT3,Survivin and COX-2 protein was correlated with lymph node metastasis (x2 =14.700,5.959,4.075,P ＜ 0.05).Overexpression of p-STAT3 was also related to neural invasion (x2=10.384,P ＜ 0.05).Expression of Survival and COX-2 protein was not associated with lymph invasion (x2=2.718,3.024,P ＞ 0.05).Expression of p-STAT3,Survivin and COX-2 was however not associated with gender,age and tumor location,differentiation and diameter (x2=0.148,0.720,1.835,1.040,0.236 ; 0.001,0.009,0.029,1.863,0.197 ; 0.433,0.686,0.002,2.974,0.029,P ＞ 0.05).Expression of Survivin and COX-2 protein was positively correlated to p-STAT3 protein (r =0.524,0.583,P ＜ 0.05).All the 43 patients were followed up for 6-60 months.Among the 17 patients with hilar cholangiocarcinoma,the median survival time was 7,9,9 months for patients with positive expression of p-STAT3,Survivin and COX-2 protein,compared with 18,11 and 11 months for patients with negative expression of these proteins.The survival rates of the patients with positive and negative expression of p-STAT3 protein were 33.3％ and 68.6％,respectively,with a statistical significance for p-STAT3 protein (x2=12.916,P ＜ 0.05).Of the remaining 26 patients with common bile duct carcinoma,the median survival time was 9,10 and 9 months for patients with positive expression of p-STAT3,Survivin and COX-2 protein,compared with 20,20 and 20 months for patients with negative expression of these proteins.The survival rates of the patients with positive expression of p-STAT3,Survivin and COX-2 protein were 20.8％,9.4％ and 8.5％,which were lower than 37.5％,37.5％ and 50.0％ of patients with the negative expression of these proteins,with a statically significance for all the 3 proteins (x2=12.787,6.245,11.161,P ＜ 0.05).Conclusions The p-STAT3,Survivin,COX-2 proteins are highly expressed in the cholangiocarcinoma and the expression levels of these proteins are positively correlated.The survivin and COX-2 may be the downstream genes of STAT signaling pathway,which are involved in the progression and prognosis of cholangiocarcinoma.

Objective To evaluate the correlation between left ventricular function and arterial stiffness of left femoral artery in patients with lower extremity atherosclerotic disease (LEAD).Methods Thirty-three patients with LEAD and 37 healthy subjects (control group) were enrolled in this study.The intima-media thickness (IMT),diameter and parameters of arterial stiffness [dispensability coefficient (DC),compliance coefficient (CC),stiffness α,stiffness β,pulse wave velocity (PWVβ) ]were measured by ultrasonography with the technology of QIMT and QAS.The thickness of the interventricular septum (IVSd),end-diastolic left ventricular diameter (LVDd) and left ventricular mass (LVM),and parameters of the left ventriculsr function (EF,E/A,E'/A',E/E' and Tei index) were measured by echocardiography.These parameters were compared between two groups.Correlations between the parameters of the arterial stiffness and those of the cardiac function were evaluated by Pearson correlative analysis.Results ①The IVSd,LVM and E/E' ratio were significantly higher in LEAD group than those in control group ( P ＜0.05).There were no significant differences in EF,E/A,E'/A',and Tei index between two groups ( P ＞0.05).②The IMT,α,β,PWVβ of left femoral artery were significantly higher in LEAD group than those in control group,while DC and CC were significantly lower in LEAD group than those in control group ( P ＜0.05).③The E/E' ratio,one of the parameters representing the left ventricular diastolic function,was correlated negatively with CC and positively with α,β,and PWVβ ( P ＜0.05 or P ＜0.01 ).The E'/A' ratio was correlated positively with DC and CC,and negatively with α,β,and PWVβ ( P ＜0.05 or P ＜0.01 ).Both EF and Tei index were not significantly correlated with the above parameters of arterial stiffness ( P ＞0.05).Conclusions Patients with LEAD have thickened femoral IMT,higher arterial stiffness of left femoral artery,as well as impaired left ventricular function.There is a close correlation between the atherosclerosis of the femoral artery and the early left vcntricular dysfunction.

Objective To evaluate the association between the left carotid arterial stiffness and left ventricular diastolic function in patients with lower extremity atherosclerosis (AS).Methods ①A total of 32 patients with AS and 34 control objects were enrolled.The carotid arterial stiffness parameters:compliance coefficient (CC),distensibility coefficient (DC),stiffness parameter (α,β),pulse wave velocity β (PWVβ) were measured by using quality arterial stiffness(QAS) technology.And the values were compared between the two groups.②The parameters of left ventricular (LV) structure and function:LV end-diastolic interventricular septal thickness (IVSd),LV end-diastolic diameter (LVDd),LV end-diastolic wall thickness (PWd),LV ejection fraction (EF),systolic velocity (s'),early-diastolic velocity (e'),Tei index and E/e' ratio were measured by using two-dimensional echocardiography and tissue Doppler.These parameters were compared between the two groups.The association between the carotid arterial stiffness parameters and LV function parameters were analyzed by correlative analysis.Results ①Compared with the control group,the DC and CC were lower,and α,β,PWVβ,IMT were higer than the control group,with statistically significant differences(P ＜0.05).②The IVSd,Tei index and E/e'was significantly higher in the AS group than those in the control group.And the PWd,s',e' were lower than those in the control group (P ＜ 0.05).There was no significant difference in EF between the two groups (P ＞0.05).③The e' was correlated positively with DC and CC (r =0.39,0.36,P ＜0.01),and negatively with α,β,and PWVβ (r =-0.42,-0.42,-0.49,P ＜0.01).Tei index was correlated negatively with DC and CC (r =-0.50,-0.52,P ＜0.01),and positively with α,β,and PWVβ (r =0.58,0.58,0.62,P ＜0.01).The E/e' was correlated regatively with CC (r =-0.27,P ＜0.05),and positively with PWVβ (r =0.28,P ＜0.05).There were no significant correlation between s',EF and the stiffness parameters of carotid artery (P＞0.05).Conclusions In patients with AS,the left carotid artery stiffness increases and left ventricular systolic and diastolic function are impaired.The carotid artery stiffness and left ventricular diastolic function is correlated.Changes in carotid artery stiffness reflect the change in left ventricular diastolic function.

ObjectiveTo investigate the role played by acute glucagon response in the short-term intensive insulin therapy induced long-term remission of newly-diagnosed type 2 diabetes.MethodsTen newly-diagnosed type 2 diabetic patients( 7 males,3 females) received intensive continuous subcutaneous insulin infusion( CSII )therapy for 2 weeks.Intravenous glucose tolerance test (IVGTT) and hyperinsulinemia euglycemia clamp test were performed before and after CSII.Glucose infusion rate ( GIR ),acute insulin response ( AIR ) and acute glucagon response (AGR) were assessed.Long-term remission was defined as good glycaemic control without any hypoglycaemic agent (fasting plasma glucose ＜ 7.0 mmol/L 及 2 h postprandial glucose ＜ 10 mmol/L) one year after CSII.Relationship between AGR and long-term remission were analyzed by one way ANOVA and Spearman correlation analysis.Results ( 1 ) Five of the ten patients achieved long-term remission in this study.GIR and AIR in the remission group were increased significantly after CSII compared with those before CSII [ GIR ( 5.39 ± 1.76 vs 2.02 ± 0.83 )mg · kg-1 ·min-1,AIR ( mean 54.1 vs mean 3.39 ) mU · L-1 · min,P＜0.01 respectively ],however both of them were not associated with the remission.(2) AGR in the remission group was significantly higher than that in the non-remission group before CSII ( 5.10±0.60 vs 2.85 ± 1.86,P＜0.05 ) and was decreased significantly after CSII.The mean of AGR after CSII was apparently lower in the remission group than that in non-remission group (0 vs 3.04±2.00,P＜0.01 ).(3)Spearman analysis showed that AGR before CSII and its range of reduction after CSII were correlated with remission ( r for both were 0.731,P=0.016).ConclusionHigher level of AGR before CSII and greater reduction after CSII in the subjects with newly-diagnosed type 2 diabetes were significantly associated with long-term remission,suggesting that pancreatic α cells may play a unique role in the induction of remission of type 2 diabetes.