The clinical characteristics of a neonatal patient with movement disorder and arthrogryposis (NDEEMA) caused by gain-of-function mutation of the SCN1A gene were reported in this article. The 1-year-and-9-month-old boy started seizures since 2 hours after birth. He had funnel-breast, dislocation of the hip, and bipedal varus. Genetic testing showed SCN1A gene de novo missense mutation c.706A>G(p.Ile236Val), causing overall gain-of-function effect. The frequency of seizures decreased significantly after treatment of oxcarbazepine.

The clinical characteristics of a neonatal patient with movement disorder and arthrogryposis (NDEEMA) caused by gain-of-function mutation of the SCN1A gene were reported in this article. The 1-year-and-9-month-old boy started seizures since 2 hours after birth. He had funnel-breast, dislocation of the hip, and bipedal varus. Genetic testing showed SCN1A gene de novo missense mutation c.706A>G(p.Ile236Val), causing overall gain-of-function effect. The frequency of seizures decreased significantly after treatment of oxcarbazepine.

Objective:To investigate the clinical features and prognosis of testicular relapse in pediatric acute lymphoblastic leukemia (ALL).Methods:Clinical data including the age, time from initial diagnosis to recurrence, relapse site, and therapeutic effect of 37 pediatric ALL with testicular relapse and treated in Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences between November 2011 and December 2022 were analyzed retrospectively. Patients were grouped according to different clinical data. Kaplan-Meier analysis was used to evaluate the overall survival (OS) rate and event free survival (EFS) rate for univariate analysis, and Cox proportional-hazards regression model was used to evaluate the influencing factors of OS rate and EFS rate for multivariate analysis.Results:The age at initial diagnosis of 37 pediatric testicular relapse patients was (5±3) years and the time from initial diagnosis to testicular recurrence was (37±15) months. The follow-up time was 43 (22, 56) months. Twenty-three patients (62%) were isolated testis relapse. The 5-year OS rate and EFS rate of the 37 relapsed children were (60±9) % and (50±9) % respectively. Univariate analysis showed that the 2-year EFS rate in the group of patients with time from initial diagnosis to testicular recurrence >28 months was significantly higher than those ≤28 months ((69±10)% vs. (11±11)%, P<0.05), 2-year EFS rate of the isolated testicular relapse group was significantly higher than combined relapse group ((66±11)% vs. (20±13) %, P<0.05), 2-year EFS rate of chimeric antigen receptor T (CAR-T) cell treatment after relapse group was significantly higher than without CAR-T cell treatment after relapse group ((78±10)% vs. (15±10)%, P<0.05). ETV6-RUNX1 was the most common genetic aberration in testicular relapsed ALL (38%, 14/37). The 4-year OS and EFS rate of patients with ETV6-RUNX1 positive were (80±13) % and (64±15) %, respectively. Multivariate analysis identified relapse occurred≤28 months after first diagnosis ( HR=3.09, 95% CI 1.10-8.72), combined relapse ( HR=4.26, 95% CI 1.34-13.52) and CAR-T cell therapy after relapse ( HR=0.15,95% CI 0.05-0.51) were independent prognostic factors for 2-year EFS rate (all P<0.05). Conclusions:The outcome of testicular relapse in pediatric ALL was poor. They mainly occurred 3 years after initial diagnosis. ETV6-RUNX1 is the most common abnormal gene.Patients with ETV6-RUNX1 positive often have a favorable outcome. Early relapse and combined relapse indicate unfavorable prognosis, while CAR-T cell therapy could significantly improve the survival rate of children with testicular recurrence.

Objective:To investigate the presence of a distinct stem cell populations different from mesenchymal stem cells in the mandibular periosteum of both human and non-human primates (macaca mulatta), to explore its properties during intramembranous osteogenesis and to establish standard protocols for the isolation, culturing and expanding of mandibular periosteal stem cells (PSC) distinguished from other PSCs in other anatomical regions.Methods:Periosteum was harvested from the bone surface during flap bone removal in patients aged 18-24 years undergoing third molar extraction and from the buccal side of the mandibular premolar region of 6-year-old macaca mulatta respectively, and then subjected to single-cell sequencing using the Illumina platform Novaseq 6000 sequencer. Cross-species single-cell transcriptome sequencing results were compared using homologous gene matching. PSC were isolated from primary tissues using two digestion methods with body temperature and low temperature, and their surface markers (CD200, CD31, CD45 and CD90) were identified by cell flow cytometry. The ability of cell proliferation and three-lineage differentiation of PSC expanded to the third generation in vitro in different species were evaluated. Finally, the similarities and differences in osteogenic properties of PSC and bone marrow mesenchymal stem cells (BMSC) were compared. Results:The single-cell sequencing results indicated that 18 clusters of cell populations were identified after homologous gene matching for dimensionality reduction, and manual cellular annotation was conducted for each cluster based on cell marker databases. The comparison of different digestion protocols proved that the low-temperature overnight digestion protocol can stably isolate PSC from the human and m. mulatta mandibular periosteum and the cells exhibited a fibroblast-like morphology. This research confirmed that PSC of human and m. mulatta had similar proliferation capabilities through the cell counting kit-8 assay. Flow cytometry analysis was then used to identify the cells isolated from the periosteum expressed CD200(+), CD31(-), CD45(-), CD90(-). Then, human and m. mulatta PSC were induced into osteogenesis, adipogenesis, and chondrogenesis to demonstrate their corresponding multi-lineage differentiation capabilities. Finally, comparison with BMSC further clarified the oesteogenesis characteristics of PSC. The above experiments proved that the cells isolated from the periosteum were peiosteal cells with characteristics of stem cells evidenced by their cell morphology, proliferation ability, surface markers, and differentiation ability, and that this group of PSC possessed characteristics different from traditional mesenchymal stem cells.Conclusions:In this study, normal mandibular PSC from humans and m. mulatta were stably isolated and identified for the first time, providing a cellular foundation for investigating the mechanism of mandibular intramembranous osteogenesis, exploring ideal non-human primate models and establishing innovative strategies for clinically mandibular injury repair.

Objective:To investigate the effect of extracellular vesicles derived from mitotic cell death PC3 cells(MEV) on prostate cancer cells.Methods:After the phenotype of mitotic death was continuously induced by 1 μmol/L cisplatin for 5 days, nuclear damage was detected by immunofluorescence, cell stiffness was measured by Atomic Force Microscope, cell senescence marker(SA-β-Gal), proliferation, cycling, mitochondrial membrane potential, mitochondrial number by flow cytometry, and expression of senescence-secreting phenotype-associated factors including CDKNIA, CDKN2A, IL-6, IL-8, TNF-α, IFN-α/β/γ by qRT-PCR. Mitotic-death PC3 cells derived extracellular vesicles (MEV) were extracted, and the morphology and size of MEV were detected by electron microscopy and nanoparticle tracking analysis, and their stiffness and substance (β-actin and mtDNA) were separately detected by atomic force microscope and qRT-PCR. The effects of MEV on PC3 cells were further detected by immunofluorescence to detect phagocytosis, flow cytometry to detect proliferation and apoptosis, and qRT-PCR to detect the level of IFNα/β/γ. Finally, 50μg MEV were treated to PC3 cells combined with 15μmol/L adriamycin (Dox), and apoptosis was detected by flow assay.Results:Mitotic cell death PC3 cells and MEV were both obtained successfully.In comparison with normal PC3's, the number of MEV group was significantly increased[(4 530.9±353.6)×10 6(particle)/1×10 6 cell and (33.7±5.4)×10 6 particle/1×10 6 cell, P<0.01]. Additionally, the particle size of the extracellular vesicles were significantly smaller[(122.0±2.6)nm and (163.6±2.6)nm, P<0.01], along with significantly increased content of nuclear DNA[(111.0±20.7)/1×10 6 cell, P<0.01] and mtDNA[(26.2±3.8)/1×10 6 cell, P<0.01]. MEV were also easier to absorb by PC3 for their softness[(0.11±0.01)MPa and(0.18±0.01)MPa, P<0.01]. MEV could significantly induce PC3 cell apoptosis[(641.0±42.5)MFI and(351.7±37.0)MFI, P<0.01] and inhibit their proliferation[(1 523.0±64.9)MFI and(1 336.3±94.1)MFI, P<0.05].Besides, they did not affect the level of IFNβ but down-regulated IFNα/γmRNA level[(0.6±0.1)and(0.8±0.1), P<0.01].The combination of MEV and 15 μmol/L Dox can significantly promote PC3 cell apoptosis[(14 290.3±1315.9)MFI and(2 669.3±241.5)MFI, P<0.01]. Conclusions:Mitotic cell death PC3 cells can efficiently secrete DNA-rich flexible extracellular vesicles.The MEV were more easily taken up by PC3 and significantly inhibit its cellular activity and promote the anticancer effect of Dox.

Objective:To investigate the effect of extracellular vesicles derived from mitotic cell death PC3 cells(MEV) on prostate cancer cells.Methods:After the phenotype of mitotic death was continuously induced by 1 μmol/L cisplatin for 5 days, nuclear damage was detected by immunofluorescence, cell stiffness was measured by Atomic Force Microscope, cell senescence marker(SA-β-Gal), proliferation, cycling, mitochondrial membrane potential, mitochondrial number by flow cytometry, and expression of senescence-secreting phenotype-associated factors including CDKNIA, CDKN2A, IL-6, IL-8, TNF-α, IFN-α/β/γ by qRT-PCR. Mitotic-death PC3 cells derived extracellular vesicles (MEV) were extracted, and the morphology and size of MEV were detected by electron microscopy and nanoparticle tracking analysis, and their stiffness and substance (β-actin and mtDNA) were separately detected by atomic force microscope and qRT-PCR. The effects of MEV on PC3 cells were further detected by immunofluorescence to detect phagocytosis, flow cytometry to detect proliferation and apoptosis, and qRT-PCR to detect the level of IFNα/β/γ. Finally, 50μg MEV were treated to PC3 cells combined with 15μmol/L adriamycin (Dox), and apoptosis was detected by flow assay.Results:Mitotic cell death PC3 cells and MEV were both obtained successfully.In comparison with normal PC3's, the number of MEV group was significantly increased[(4 530.9±353.6)×10 6(particle)/1×10 6 cell and (33.7±5.4)×10 6 particle/1×10 6 cell, P<0.01]. Additionally, the particle size of the extracellular vesicles were significantly smaller[(122.0±2.6)nm and (163.6±2.6)nm, P<0.01], along with significantly increased content of nuclear DNA[(111.0±20.7)/1×10 6 cell, P<0.01] and mtDNA[(26.2±3.8)/1×10 6 cell, P<0.01]. MEV were also easier to absorb by PC3 for their softness[(0.11±0.01)MPa and(0.18±0.01)MPa, P<0.01]. MEV could significantly induce PC3 cell apoptosis[(641.0±42.5)MFI and(351.7±37.0)MFI, P<0.01] and inhibit their proliferation[(1 523.0±64.9)MFI and(1 336.3±94.1)MFI, P<0.05].Besides, they did not affect the level of IFNβ but down-regulated IFNα/γmRNA level[(0.6±0.1)and(0.8±0.1), P<0.01].The combination of MEV and 15 μmol/L Dox can significantly promote PC3 cell apoptosis[(14 290.3±1315.9)MFI and(2 669.3±241.5)MFI, P<0.01]. Conclusions:Mitotic cell death PC3 cells can efficiently secrete DNA-rich flexible extracellular vesicles.The MEV were more easily taken up by PC3 and significantly inhibit its cellular activity and promote the anticancer effect of Dox.

Objective To investigate the correlation between the proportion of peripheral blood follicular T helper cells(Tfh cells)and intracellular interleukin-21(IL-21)content with blue light retinal injury.Methods Brown Norway(BN)rats were randomly divided into 4 groups and were exposed to blue light for 3 hours a day to establish retinal light damage model.According to the duration of illumination,the rats was divided into 0 days(control group),3 days(3 d group),7 days(7 d group)and 14 days(14 d group).The proportion of Tfh cells and content of IL-21 in Tfh cells in peripheral blood of each group was detected by flow cytometry and ELISA sepa-rately after illumination.Electroretinogram(ERG)was used to evaluate retinal function.The changes of fundus in rats were observed by fundus photography.The thickness of outer nuclear layer of retina was analyzed by HE staining.Results After retinal blue light injury,with the extension of illumination time,the proportion of Tfh cells in peripheral blood and intracellular IL-21 content both increased(P<0.05).ERG showed that retinal function decreased after light damage and aggravated with the extension of illumination time,the latency and ampli-tudes of A-wave and B-wave increased and decreased respectively(P<0.05).The retinal fundus of rats showed depigmentation in 3 d,and the retinal vessels became thinner and exudate with the extension of illumination time.HE staining showed that the outer nuclear layer of retina(ONL)became thinner(P<0.05).Correlation analy-sis indicated that the proportion of Tfh cells in peripheral blood and the intracellular IL-21 content could jointly reflect the degree of injury(P<0.000 1),and the proportion of Tfh cells in peripheral blood was negatively corre-lated with ONL thickness and the amplitude of a and b waves,positively correlated with the peak time of a and b waves,(P<0.0001).Conclusion The proportion of Tfh cells in peripheral blood and the intracellular IL-21 content were increased after blue light damage to retina,and were significantly increased with the extension of light time with a certain correlation.

BACKGROUND: Immunotherapies such as adoptive immune cell infusion and immune-modulating agents are widely used for cancer treatment, and the concomitant symptoms, including cytokine release syndrome (CRS) or immune-related adverse events (irAEs), are frequently reported. However, clinical manifestations induced by mismatched donor granulocyte colony-stimulating factor mobilized peripheral blood mononuclear cell (GPBMC) infusion in patients receiving microtransplant (MST) have not yet been well depicted. METHODS: We analyzed 88 cycles of mismatched GPBMC infusion in patients with acute myeloid leukemia receiving MST and 54 cycles of chemotherapy without GPBMC infusion as a comparison. Clinical symptoms and their correlation with clinical features, laboratory findings, and clinical response were explored. RESULTS: Fever (58.0% [51/88]) and chills (43.2% [38/88]) were the significant early-onset symptoms after GPBMC infusion. Patients possessing less human leukocyte antigen-matching loci with the donor or those with unrelated donors experienced more chills (3 [2-5] loci vs. 5 [3-5] loci, P  = 0.043 and 66.7% [12/18] vs. 37.1% [26/70], P  = 0.024). On the other hand, those with decreased CD4 + /CD8 + T-cell ratio developed more fever (0.8 [0.7-1.2] vs. 1.4 [1.1-2.2], P  = 0.007). Multivariable analysis demonstrated that younger patients experienced more fever (odds ratio [OR] = 0.963, 95% confidence interval [CI]: 0.932-0.995, P  = 0.022), while patients with younger donors experienced more chills (OR = 0.915, 95% CI: 0.859-0.975, P  = 0.006). Elevated ultra-sensitive C-reactive protein levels in the absence of cytokine storm were observed following GPBMC infusion, which indicated mild and transient inflammatory response. Although no predictive value of infusion-related syndrome to leukemia burden change was found, the proportion of host pre-treatment activated T cells was positively correlated with leukemia control. CONCLUSIONS Mismatched GPBMC infusion in MST induced unique infusion-related symptoms and laboratory changes, which were associated with donor- or recipient-derived risk factors, with less safety and tolerance concerns than reported CRS or irAEs.
Humans ; Leukocytes, Mononuclear ; Hematopoietic Stem Cell Transplantation/adverse effects* ; Leukemia, Myeloid, Acute/therapy* ; Unrelated Donors ; Granulocyte Colony-Stimulating Factor ; Graft vs Host Disease

Objective:To investigate the relationship between arterial blood partial pressure of carbon dioxide and neurological outcome after cardiopulmonary resuscitation.Methods:The clinical data of 116 patients who underwent cardiopulmonary resuscitation admitted to the Intensive Care Unit and Emergency Department of the Second People's Hospital of Hefei from January 2018 to January 2020 were retrospectively analyzed. According to the average arterial blood partial pressure of carbon dioxide within 24 hours after admission, patients were divided into normal (35 mmHg ≤ PaCO 2 ≤ 55 mmHg, 1 mmHg = 0.133 kPa, n = 44), hypercapnia (PaCO 2 > 55 mmHg, n = 51), and hypocapnia (PaCO 2 < 35 mmHg, n = 21) groups. ICU stay, in-hospital mortality, and neurological outcome at discharge were compared among groups. A logistic regression analysis model was established. The relationship between PaCO 2 and neurological outcome was determined. Results:There were no significant differences in age, sex, cardiac arrest time, acute physiological and chronic health evaluation II score at admission, 1-hour mean arterial pressure, location of cardiac arrest, and initial heart rhythm among the three groups (all P > 0.05). ICU stay in the normal group [(7.23 ± 2.55) days] was significantly higher than that in the hypercapnia [(12.21 ± 4.12) days] and hypocapnia [(11.78 ± 4.72) days] groups ( t = 6.48, 4.59, both P < 0.01). In-hospital mortality in the normal group was 38.6% (17/44), which was significantly lower than 60.8% (31/51) in the hypercapnia group and 66.7% (14/21) in the hypocapnia group ( χ2 = 4.63, 4.47, both P < 0.05). The good neurological outcome rate in the normal group was 55.6% (15/44), which was significantly higher than 25.0% (5/51) in the hypercapnia group and 28.6% (2/21) in the hypocapnia group ( χ2 = 8.38, 5.14, both P < 0.05). Multivariate logistic regression analysis showed that cardiac arrest time, 1-hour mean arterial pressure, acute physiological and chronic health evaluation II score, and PaCO 2 are important factors for neurological outcomes of resuscitated patients at discharge (all P < 0.01). Conclusion:Within 24 hours after cardiopulmonary resuscitation, maintaining a normal PaCO 2 level can help improve the neurological outcome of patients at discharge.

Long non-coding RNAs (lncRNAs) play a significant role in maintaining tissue morphology and functions, and their precise regulatory effectiveness is closely related to expression patterns. However, the spatial expression patterns of lncRNAs in humans are poorly characterized. Here, we constructed five comprehensive transcriptomic atlases of human lncRNAs covering thousands of major tissue samples in normal and disease states. The lncRNA transcriptomes exhibited high consistency within the same tissues across resources, and even higher complexity in specialized tissues. Tissue-elevated (TE) lncRNAs were identified in each resource and robust TE lncRNAs were refined by integrative analysis. We detected 1 to 4684 robust TE lncRNAs across tissues; the highest number was in testis tissue, followed by brain tissue. Functional analyses of TE lncRNAs indicated important roles in corresponding tissue-related pathways. Moreover, we found that the expression features of robust TE lncRNAs made them be effective biomarkers to distinguish tissues; TE lncRNAs also tended to be associated with cancer, and exhibited differential expression or were correlated with patient survival. In summary, spatial classification of lncRNAs is the starting point for elucidating the function of lncRNAs in both maintenance of tissue morphology and progress of tissue-constricted diseases.
Humans ; Gene Expression Profiling ; Neoplasms/genetics* ; Organ Specificity ; RNA, Long Noncoding/genetics* ; Transcriptome