Objective: In in vitro systems for differentiating and expanding natural killer (NK) cells, feeder cells provide essential cell-cell contact and paracrine signals that drive precursor proliferation and terminal maturation. However, existing xenogeneic feeder cells or tumor-derived genetically modified feeder cells pose risks of residual immunogenicity and malignant transformation, limiting clinical use. This study aims to develop a humanized mesenchymal-like stromal cell (hematopoietic organoid-derived human mesenchymal stromal cells, HO-hMSCs) derived from iPSC-based hematopoietic organoids, and elucidate its mechanisms of NK-supportive activity to enable a safe, efficient platform for clinical-grade NK cell production. Methods: Human induced pluripotent stem cells (iPSCs) were differentiated into hematopoietic organoids, from which HO-hMSCs were isolated. Flow-cytometric phenotyping and bulk RNA-sequencing were performed to compare HO-hMSCs with umbilical cord-derived MSCs (UC-hMSCs). The effect of HO-hMSCs on NK cell differentiation efficiency (CD3 CD56 ) and effector maturation (CD16 expression) were assessed by co-culture experiments, using UC-hMSCs as control. Results: 1) Hematopoietic organoid induction and NK differentiation: iPSCs were induced to form hematopoietic organoids using cytokine cocktails, which further differentiated into high-purity CD45 CD56 NK cells [(82.8%±12.07)% efficiency on day 21]. 2) HO-hMSC characteristics: HO-hMSCs exhibited upregulated expression of Notch pathway ligands (DLL4, JAG1, 4.06-8.04-fold), homeobox genes (HOXA3, HOXA5, log FC=1.28 and 1.44), and key regulators of NK development (GATA3, BCL11A) and cytokine receptors (IL7R, IL27RA, 6.76 to 13.34-fold increase). 3) Functional validation: Compared to UC-hMSCs, HO-hMSCs co-culture significantly enhanced NK cell proportion by 30.5% (P<0.05) and increased CD16 positivity (+20.5%). Conclusion: This study for the first time reveals that human hematopoietic organoid-derived HO-hMSCs possess potent hematopoietic niche-supportive activity. It provides a humanized, feeder-free platform for robust clinical-grade NK cell production and expands the translational utility of organoid technologies in cell therapy.

OBJECTIVE: To investigate the transfusion and infusion warmer manufacturers, combine the use failures to analyze adverse events, and provide support for enterprise risk management and clinical safe use. METHODS: The sentinels from 7 manufacturing enterprises and 11 medical institutions that participated in Shandong Province's key monitoring program during the "14th Five-Year Plan" period were targeted. This was done by understanding the equipment's principles, structures, and quality control. Additionally, real-world data from January 2019 to December 2023 were collected to count adverse events. RESULTS: During production, there are risks in switching power supply stability and solder joint firmness. Fifteen kinds of faults occurred during use, and common faults such as inability to heat, unable to turn on the machine, and bubbles in the infusion tube accounted for more than 80%. CONCLUSION There are many risk points and failures for transfusion and infusion warmers, so enterprises should improve processes and quality control to address risks, and medical institutions should formulate specifications and maintenance plans to provide targeted theoretical basis for supervision.
Blood Transfusion/instrumentation* ; Risk Factors ; Quality Control ; Humans ; Risk Management ; Equipment Failure

OBJECTIVES: To analyze the differentially expressed exosomal miRNAs in patients with chronic heart failure (CHF) complicated by hyperuricemia (HUA) and explore their potential as novel diagnostic molecular markers and their target genes. METHODS: This study was conducted among 30 CHF patients with HUA (observation group) and 30 healthy volunteers (control group) enrolled between September, 2020 and September, 2023. Peripheral blood samples were collected from 6 CHF patients with HUA for analyzing exosomal miRNAs by high-throughput sequencing, and the results were validated in the remaining 24 patients using qRT-PCR. GO and KEGG enrichment analyses were performed to predict the the target genes of the identified differential miRNAs. We also validated the differentially expressed miRNAs by animal experiment. RESULTS: A total of 42 differentially expressed exosomal miRNAs were detected in observation group by high-throughput sequencing; among them, miR-27a-5p was significantly upregulated (P=0.000179), and miR-139-3p was significantly downregulated (P=0.000058). In the 24 patients with both CHF and PUA, qRT-PCR validated significant upregulation of miR-27a-5p (P=0.004) and downregulation of miR-139-3p (P=0.005) in serum exosomes. When combined, miR-27a-5p and miR-139-3p had a maximum area under the curve (AUC) of 0.899 (95% CI: 0812-0.987) for predicting CHF complicated by HUA. GO and KEGG enrichment analyses suggested that the differential expressions of miR-27a-5p and miR-139-3p was associated with the activation of the AMPK-mTOR signaling pathway to activate the autophagic response. We obtained the same conclusion from animal experiment. CONCLUSIONS Upregulated exosomal miR-27a-5p combined with downregulated exosomal miR-139-3p expression can serve as a novel molecular marker for diagnosis of CHF complicated by HUA, and their differential expression may promote autophagy in cardiomyocytes by activating the AMPK-mTOR signaling pathway.
Humans ; Hyperuricemia/diagnosis* ; Heart Failure/genetics* ; MicroRNAs/metabolism* ; Exosomes/metabolism* ; Chronic Disease ; Male ; Female ; Middle Aged ; Animals

Objective Utilizing a specially engineered miR-144-GFP-H1 human embryonic stem cell(hESC)reporter line,this study leverages GFP fluorescence as an indicator of miR-144 expression to gauge the progression of erythropoiesis.The investigation is aimed at elucidating the potential roles of lncRNAs within the erythropoietic framework and conducting an initial assessment of their functional impact.Methods The miR-144/451-GFP-H1 cell line(hereafter referred to as 144-H1)was utilized for in vitro erythrocyte induction culture.The subpopulations of cells entering the erythropoiesis stage were characterized by the surface molecules CD71 and GPA.The GFP reporter gene of miR-144 served as a critical determi-nant to distinguish between GFP-positive cells(with a high propensity for erythropoiesis)and GFP-negative cells(with a low propensity for erythropoiesis).Transcriptome sequencing was performed on both groups to identify differentially ex-pressed long non-coding RNAs(lncRNAs).LncRNA entries with potential for validation were selected for preliminary func-tional verification.The CRISPR/Cas9 gene editing technique was employed to design functional interference strategies for the targeted lncRNAs,obtaining 144-H1 cell lines with knocked-out function of the specific lncRNAs.These knockout cell lines,along with non-knockout 144-H1 cell lines,were used for parallel erythrocyte induction culture to identify differential nodes.This approach preliminarily verified their impact on erythropoiesis in an in vitro development model.Results 1)The constructed 144-H1 cell line was capable of expressing GFP fluorescence upon entering the stage of in vitro erythrocyte in-duction,indicating the activation of miR-144/451.2)Within the CD71,GPA double-positive group,significant differences in lncRNA expression were observed between the GFP-positive and GFP-negative subpopulations.3)Gene editing strategies involving the deletion of sequence segments capable of effectively interfering with the function of multiple lncRNA entries were designed and verified for successful editing.In the knockout cell lines,parts of the lncRNA sequences were directly de-leted.4)In parallel validation experiments of erythrocyte induction culture,cell lines with LINC01569 knocked out exhibited significant differences in flow cytometric subpopulations and cell proliferation capabilities compared to the non-knockout cell lines:①The knockout cell lines showed sustained high expression of GFP fluorescence.②The proportion of the CD71-GPA double-positive group in the knockout cell lines continuously decreased during erythrocyte maturation.③No significant ex-pression of hemoglobin was observed in the knockout cell lines,lacking the characteristic red color.④The cell proliferation capability of the knockout cell lines was significantly lower than that of the non-knockout cell lines(P＜0.05).Conclusion The successful employment of the 144-H1 cell line facilitated an exploration into the potential functions of lncRNAs in e-rythropoiesis.This enables the design of more refined in vitro developmental experiments to enhance the precision in captu-ring lncRNA functions.Among the differentially expressed lncRNA entries,LINC01569 was preliminarily validated to play a regulatory role in erythropoiesis.The functional absence of LINC01569 severely impacts the normal differentiation and prolif-eration of erythrocytes.The specific regulatory mechanism of LINC01569 in erythropoiesis warrants further investigation and research.

Objective:To investigate the effectiveness of rotator cuff repair surgery in treating proximal humeral fractures and its association with patient age and Neer classification.Methods:A retrospective analysis was conducted on the clinical data of 90 patients with proximal humeral fractures who underwent rotator cuff repair surgery at the Marine Police Corps Hospital of the Chinese People's Armed Police Force between January 2020 and January 2023. The clinical data of these patients were retrieved from the hospital's medical record database. Based on the age distribution of the patients, they were divided into four groups: Group 1 (> 18 years old and < 30 years old, n = 10), Group 2 (≥ 30 years old and < 45 years old, n = 15), Group 3 (≥ 45 years old and < 60 years old, n = 40), and Group 4 (> 60 years old, n = 25). There were no statistically significant differences in baseline data among the four groups (all P > 0.05), indicating comparability. Additionally, these patients were divided into three groups based on Neer classification: Group I (Neer type II, n = 28), Group II (Neer type III, n = 30), and Group III (Neer type IV, n = 32). Differences in various outcome measures related to treatment efficacy, including operation time, intraoperative blood loss, fracture healing time, Visual Analogue Scale (VAS) score, Constant-Murley score, Neer shoulder function score, and the overall incidence of postoperative complications, were recorded and compared across different age groups. Overall efficacy was assessed according to the Neer criteria: > 90 points as excellent, 80-89 as good, 70-79 as fair, and < 70 as poor. Results:After 6 months of surgery, the overall clinical treatment effectiveness rate was 95.56% (86/90). At 2 weeks postoperatively, the VAS score was (3.45 ± 1.35) points, which was significantly decreased compared with the level measured preoperatively ( P < 0.05). At 6 months postoperatively, the Constant-Murley score and Neer shoulder function score were (90.34 ± 6.34) points and (86.34 ± 5.46) points, respectively (both P < 0.05), which were significantly increased compared with the levels measured preoperatively. There were significant differences in fracture healing time ( F = 4.89, P < 0.05), intraoperative blood loss ( F = 5.28, P < 0.05), VAS score ( F = 5.02, P < 0.05), and Neer shoulder function score ( F = 5.32, P < 0.05) and Constant-Murley scores ( F = 3.85, P < 0.05) compared with before treatment. There was no statistically significant difference in the overall incidence of postoperative complications or surgical time among Groups 1, 2, 3,and 4 [0 vs. 5.00% (2/40) vs. 6.67% (1/15) vs. 12.00% (3/25), χ2 = 0.98, P > 0.05]. Group III had longer surgical time and fracture healing time compared with Groups I and II ( F = 4.55, 4.23, both P < 0.05), with greater intraoperative blood loss ( F = 5.24, both P < 0.05). Additionally, Group III had a higher VAS score compared with Groups I and II ( F = 5.06, P < 0.05), while the Neer shoulder joint function score and Constant-Murley score in Group III were lower ( F = 3.83, 4.56, both P < 0.05). There was no statistically significant difference in the overall incidence of postoperative complications among the Groups I, II, and III [9.38% (3/32) vs. 3.57% (1/28) vs. 6.67% (2/30), χ2 = 1.00, all P > 0.05]. Pearson correlation analysis revealed a positive correlation between patient age, Neer classification, intraoperative blood loss, fracture healing time, and VAS score ( r = 0.565, all P < 0.05), and a negative correlation with the Neer shoulder joint function score ( r = -0.620, P < 0.05). However, there was no significant correlation between patient age, Neer classification, and the Constant-Murley score ( r = -0.008, P > 0.05). Conclusion:The rotator cuff repair is effective in treating proximal humeral fractures, but the effectiveness varies depending on patient age and Neer classification. Patient age and Neer classification are positively correlated with intraoperative blood loss, fracture healing time, and VAS score, while they are negatively correlated with the Neer shoulder joint function score.

Objective:To investigate the effect of chimeric flap pedicled with superficial branch of superficial iliac circumflex artery in repair of soft tissue defect of dorsal hand combined with metacarpal bone defect.Methods:From May 2015 to January 2022, 34 patients(28 males and 6 females) of soft tissue defects of dorsal hand with metacarpal bone defects were treated in the Department of Orthopedics of Yibin Third People's Hospital. The age of patients ranged from 22 to 51 years old, with an average age of 37 years old. The areas of soft tissue defects after debridement were 2.5 cm×5.0 cm-4.5 cm×9.0 cm, and the defects were all in dorsal hand and dorsal wrist. The lengths of metacarpal bone defect were 1.8-4.1 cm. All the patients had only single metacarpal bone defect, among which: 14 patients had defects in first metacarpal bone, 7 in second metacarpal bone, 4 in third metacarpal bone, 8 in fourth metacarpal bone and 1 in fifth metacarpal bone. All the patients were repaired by chimeric flap pedicled with superficial branch of superficial iliac circumflex artery. The size of flaps were 3.6 cm×5.4 cm-5.2 cm×9.5 cm. Anticoagulation, thermal preservation and plaster fixation were applied for 4-6 weeks after surgery. Postoperative follow-ups included regularly outpatient clinic visit, telephone or Wechat reviews. Follow-up items covered: the feeling and appearance of flaps in recipient sites, healing of the donor sites and recovery of hand functions.Results:All the 34 chimeric flaps survived. Regular follow-up lasted for 3 to 15(average, 10) months. All incisions in the donor sites of hip healed in stage I. TPD of the flaps was 5.1-7.3(mean, 6.4) mm. Appearance of flaps in the receiving area were satisfactory without swelling. Movement of wrists and metacarpophalangeal joints met the basic requirement of movement. The healing time of metacarpal defect was 2-3 months with an average of 2.8 months. Hand functions were evaluated at excellent in 6 patients and good in 28, according to the Evaluation Standard of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association.Conclusion:The chimeric flap pedicled with superficial branch of superficial iliac circumflex artery is an ideal flap to repair the soft tissue defect in dorsal hand combined with metacarpal bone defect. It has advantages of less donor site damage, good blood supply of flap, simple surgical procedure, and one-stage repair of a combined soft tissue and metacarpal bone defects.

Objective:To evaluate whether FOXO4-DRI could reverse radiation-induced pulmonary fibrosis (RIPF) and to explore the underlying mechanism.Methods:C57BL/6 mice were randomly divided into 4 groups: control, FOXO4-DRI, radiation, and radiation+ FOXO4-DRI. Mice in radiation or radiation+ FOXO4-DRI groups received 17 Gy X-ray radiation on the right side of the whole chest. Mice in FOXO4-DRI and radiation+ FOXO4-DRI groups were injected with FOXO4-DRI intraperitoneally at 16 and 20 weeks after irradiation, respectively. The right lungs were collected at 24 weeks after irradiation and subjected to HE staining and Masson trichrome staining to observe the morphological changes and collagen deposition. Immunohistochemistry was used to evaluate the expressions of col1α1 and α-SMA in lung tissues. β-gal staining was used to observe senescent cells. The level of reactive oxygen species in lung tissue was detected. The expressions of P21, P16 Ink4a and senescence-associated secretory phenotype (SASP) mRNA were detected by qRT-PCR, and the expression of related proteins were assessed by Western blot. Results:FOXO4-DRI reduced collagen deposition ( t=6.18, P<0.05), down-regulated the expression of col1α1 and α-SMA ( t=4.69, 3.20, P<0.05), and reduced the number of β-gal positive cells ( t=6.09, P<0.05) in the lung tissue of RIPF mice. FOXO4-DRI also down-regulated the gene and protein expressions of P21 and P16 Ink4a ( t=5.31, 3.32 and 4.77, 3.37, P<0.05) and inhibited the expressions of SASP genes IL-1α, IL-1β, TNF-α and MMP2 ( t=4.36, 4.84, 4.47, 3.82, P<0.05), reduced reactive oxygen species ( t=2.84, P<0.05), and promoted the activation of p-AKT and p-PI3K proteins ( t=-7.13, -12.61, P< 0.05) in the lung tissue of RIPF mice. Conclusions:FOXO4-DRI reverses RIPF by activating the PI3K/AKT signaling pathway, reducing oxidative stress and inhibiting cellular senescence.

Lobectomy is the standard surgical procedure of non-small cell lung cancer (NSCLC). Based on parenchymal-sparing advantage, better postoperative lung function, rapid recovery and less invasiveness, segmentectomy has been widely used in early peripheral non-small cell carcinoma in recent years. But there was no randomized clinical trials confirming survival benefit of segmentectomy. Led by Asamura, the Japanese Clinical Oncology Group (JCOG) has conducted a series of studies on this topic. Ever since the presentation at the 101st Annual Meeting of the American Society of Thoracic Surgeons (AATS) in 2021, the results of JCOG0802/WJOG4607l have triggered massive debate. This study was aimed at determining whether segmentectomy was non-inferior to lobectomy in overall survival in patients with early peripheral NSCLC (tumor diameter≤2 cm and consolidation tumor ratio>0.5), and the results were published in The Lancet on 22 April 2022. The 5-year overall survival rate was higher in the segmentectomy group than that in the lobectomy group, despite a higher rate of local recurrence, suggesting that segmentectomy should be the standard surgical procedure for those patients. Results of this study provide high-level evidence-based medicine evidence for the safety and effectiveness of segmentectomy, and are expected to promote the application of segmentectomy in those patients or even more other patient populations. However, due to the increased local recurrence rate and unsatisfactory postoperative lung function, there are still problems to be solved to make segmentectomy a standard surgical procedure. This paper interprets this study, discusses its instructiveness in clinical practice and summarizes its limitations.

Objective:To evaluate the characteristics of carotid plaque and the immediate outcomes after carotid artery stenting (CAS) in diabetic and non-diabetic patients by optical coherence tomography (OCT).Methods:Patients underwent CAS and OCT before and after operation in the Department of Neurology, Jinling Hospital from January 2014 to March 2019 were enrolled retrospectively. The clinical features, the characteristics of carotid plaque on OCT and the immediate outcomes after CAS were compared between diabetic group and non-diabetic group. The risk factors of stent malapposition were analyzed.Results:A total of 46 patients were enrolled. Their age was 64.02±8.32 years and 41 were males (89.1%). There were 20 patients (43.5%) in the diabetes group and 26 (56.5%) in the non-diabetes group. The proportions of atherosclerotic plaque with thin fibrous cap (40.0% vs. 7.7%; χ2=5.166, P=0.023), plaque rupture (55.0% vs. 23.1%; χ2=4.945, P=0.026) and macrophage infiltration (60.0% vs. 30.8%; χ2=3.930, P=0.047) in the diabetic group were significantly higher than those in the non-diabetic group. Multivariate logistic regression analysis showed that older age (odds ratio [ OR] 1.208, 95% confidence interval [ CI] 1.033-1.413; P=0.018), coronary heart disease ( OR 15.953, 95% CI 1.142-222.952; P=0.040), alcohol consumption ( OR 6.192, 95% CI 1.098-34.923; P=0.039) and lower systolic blood pressure ( OR 0.944, 95% CI 0.894-0.997; P=0.037) were independently associated with stent malaposition. Conclusion:Compared with the non-diabetic patients, carotid plaque in diabetic patients may be more unstable. Older age, coronary heart disease, alcohol consumption and lower systolic blood pressure were associated with stent malaposition after carotid stenting. OCT can reveal the characteristics of carotid plaque and the immediate outcomes after CAS, which can provide strong evidence for treatment decision.

Objective:To analyze the survival benefits and treatment related toxic effects of simultaneous integrated boost intensity-modulated radiotherapy (SIB-RT) for non-operative esophageal squamous cell carcinoma patients.Methods:The data of 2 132 ESCC patients who were not suitable for surgery or rejected operation, and underwent radical radiotherapy from 2002 to 2016 in 10 hospitals of Jing-Jin-Ji Esophageal and Esophagogastric Cancer Radiotherapy Oncology Group (3JECROG) were analyzed. Among them, 518 (24.3%) cases underwent SIB (SIB group) and 1 614 (75.7%) cases did not receive SIB (No-SIB group). The two groups were matched with 1∶2 according to propensity score matching (PSM) method (caliper value=0.02). After PSM, 515 patients in SIB group and 977 patients in No-SIB group were enrolled. Prognosis and treatment related adverse effects of these two groups were compared and the independent prognostic factor were analyzed.Results:The median follow-up time was 61.7 months. Prior to PSM, the 1-, 3-, and 5-years overall survival (OS) rates of SIB group were 72.2%, 42.8%, 35.5%, while of No-SIB group were 74.3%, 41.4%, 31.9%, respectively ( P=0.549). After PSM, the 1-, 3-, and 5-years OS rates of the two groups were 72.5%, 43.4%, 36.4% and 75.3%, 41.7%, 31.6%, respectively ( P=0.690). The univariate survival analysis of samples after PSM showed that the lesion location, length, T stage, N stage, TNM stage, simultaneous chemoradiotherapy, gross tumor volume (GTV) and underwent SIB-RT or not were significantly associated with the prognosis of advanced esophageal carcinoma patients who underwent radical radiotherapy ( P<0.05). Cox model multivariate regression analysis showed lesion location, TNM stage, GTV and simultaneous chemoradiotherapy were independent prognostic factors of advanced esophageal carcinoma patients who underwent radical radiotherapy ( P<0.05). Stratified analysis showed that, in the patients whose GTV volume≤50 cm 3, the median survival time of SIB and No-SIB group was 34.7 and 30.3 months ( P=0.155), respectively. In the patients whose GTV volume>50 cm 3, the median survival time of SIB and No-SIB group was 16.1 and 20.1 months ( P=0.218). The incidence of radiation esophagitis and radiation pneumonitis above Grade 3 in SIB group were 4.3% and 2.5%, significantly lower than 13.1% and 11% of No-SIB group ( P<0.001). Conclusions:The survival benefit of SIB-RT in patients with locally advanced esophageal carcinoma is not inferior to non-SIB-RT, but without more adverse reactions, and shortens the treatment time. SIB-RT can be used as one option of the radical radiotherapy for locally advanced esophageal cancer.