Objective To investigate the expression profile of cell adhesion molecular 3(CADM3)on pulmonary endothelial cells,analyze its role in mediating specific adhesion to monocyte,and explore a new mechanism of hypoxia inducing peripheral monocyte infiltration in pulmonary vessels.Methods Human umbilical vein endothelial cells(HUVEC),rat pulmonary vascular endothelial cells(rPEC),and rat aortic endothelial cells(rAEC)were subjected,and divided into normoxic(21%O2)control group and hypoxic(1%O2 or 5%O2)treatment group.Analyzing the transcriptome data of HUVEC exposure to hypoxia for 8 h screened a differentially expressed molecule,CADM3.Cell adhesion experiments,siRNA interference,immunohistochemical assay,Western blotting,and flow cytometry were used to study the role of CADM3 in hypoxia specific high adhesion of HUVEC monocytes.Iron chelator deferoxamine(DFX)and shRNA interference were employed to enhance the expression of HIF-1α,and the effect of HIF-1 transcriptional activity on CADM3 expression was analyzed.Results Hypoxic treatment resulted in enhanced expression of CADM3 in HUVEC,and the expression level reached the peak at 6-8 h after hypoxia,and then decreased.Transfection with siRNA targeting CADM3 decreased the expression of CADM3,and significantly reduced the adhesion rate of hypoxic HUVEC-U937 cells when compared with the negative control group(P<0.05).Compared with the solvent control group,the protein levels of HIF-1α and its target protein STC2 in HUVEC treated with DFX(100 μmol/L)were increased.Transfection with shRNA targeting HIF-1α led the protein levels of HIF-1α and its target protein STC2 decreased,but had no effect on the protein expression of CADM3 in comparison to the negative control group.The protein level and distribution of CADM3 on the cell membrane were increased in hypoxic rPEC.No expression of CADM3 protein was found in the rAEC when compared with rPEC.After treatment with 5 μg/mL LPS,there were no significant changes in HIF-1α,STC2 and CADM3 in rPEC cultured under normoxia or hypoxia.The adhesion rate between hypoxia rPEC with CD11b+cells was the highest,with statistical significance(P<0.01).After incubation with anti-CADM3 antibody,the adhesion of hypoxic rPEC-U937 was significantly decreased compared with the solvent control group(P<0.05).Conclusion Hypoxia specifically induces the expression of CADM3 in pulmonary vascular endothelial cells in a HIF-1-independent manner,and promotes the specific adhesion of pulmonary vascular endothelial cells and monocytes induced by hypoxia.

Objective To explore the role of X chromosome encoded epigenetic regulator UTX in NK cell-mediated anti-tumor activity.Methods Male Ncr1-iCre mice were crossed with female UTXfl/fl mice to generate F1 Ncr1-iCre+UTXfl/-male mice,which were further crossed with female UTXfl/fl mice to obtain male Ncr1-iCre-UTX fl/-control mice(M-Con)and NK-specific deletion of UTX male mice Ncr1-iCre+UTXfl/-(M-KO),as well as female Ncr1-iCre-UTXfl/fl control mice(F-Con)and UTX-deficient female mice Ncr1-iCre+UTXfl/fl(F-KO).UTX-deficient mice were injected with melanoma cell line B16F10 via tail vein to observe pulmonary metastatic tumor nodules.Moreover,flow cytometry was applied to detect the proportion and quantity of pulmonary NK cells(CD3-CD19-NK1.1+),maturation makers KLRG1 and CD11b,activation receptors NKG2D and CD69,and effector molecules,including perforin,granzyme B,CD107a,and IFN-γ.Then pulmonary NK cells were sorted and co-cultured with B16F10 cells,and the apoptosis of the melanoma cells was measured with flow cytometry.Results Compared with the M-Con mice,the M-KO mice presented less number of pulmonary tumor nodules(P<0.05),increased proportion and quantity of NK cells in the tumor microenvironment(P<0.01),though no obvious changes in the ratio of NK maturation makers KLRG1 to CD11b,enhanced expression level of cytotoxic molecule perforin(P<0.01),but no changes in the expression of effector molecule granzyme B,degranulation marker CD107a and cytokine IFN-γ in NK cells.Co-culture of NK cells and B16F10 cells promoted the apoptosis of tumor cells(P<0.05).Compared with the F-Con mice,the F-KO mice had no statistical difference in the number of pulmonary tumor nodules,but larger proportion and number of NK cells(P<0.05),decreased ratio of KLRG1 to CD11b(P<0.01),elevated level of perforin but decreased levels of granzyme B,CD107a and IFN-γ in NK cells(P<0.01).The co-culture of NK cells and B16F10 cells reduced the apoptosis of tumor cells in F-KO female mice(P<0.05).Conclusion NK-specific deletion of UTX regulates pulmonary metastasis of melanoma in a sex-dependent manner.

Acute hypobaric hypoxic brain damage is a potentially fatal high-altitude sickness. Autophagy plays a critical role in ischemic brain injury, but its role in hypobaric hypoxia (HH) remains unknown. Here we used an HH chamber to demonstrate that acute HH exposure impairs autophagic activity in both the early and late stages of the mouse brain, and is partially responsible for HH-induced oxidative stress, neuronal loss, and brain damage. The autophagic agonist rapamycin only promotes the initiation of autophagy. By proteome analysis, a screen showed that protein dynamin2 (DNM2) potentially regulates autophagic flux. Overexpression of DNM2 significantly increased the formation of autolysosomes, thus maintaining autophagic flux in combination with rapamycin. Furthermore, the enhancement of autophagic activity attenuated oxidative stress and neurological deficits after HH exposure. These results contribute to evidence supporting the conclusion that DNM2-mediated autophagic flux represents a new therapeutic target in HH-induced brain damage.
Mice ; Animals ; Hypoxia ; Oxidative Stress ; Autophagy ; Cognition ; Sirolimus/therapeutic use*

Objective:To analyze the clinical and epidemiological characteristics of adult carotid body tumors (CBTs) in Northwest China to provide references for early diagnosis and treatment of CBTs.Methods:A multicenter, retrospective, non-intervention epidemiological investigation was conducted on adult CBTs patients who were hospitalized from January 1, 2011 to June 30, 2023 in 7 Class A tertiary hospitals in Northwest China (Departments of Neurosurgery, First Affiliated Hospital of Air Force Medical University, Second Affiliated Hospital of Lanzhou University, People's Hospital of Gansu Province, 940 th Hospital of PLA Joint Logistic Support Force, People's Hospital of Qinghai Province, General Hospital of Ningxia Medical University, People's Hospital of Ningxia Hui Autonomous Region). Medical records were collected in these patients, and they were divided into 2 groups according to their average altitude residence: high altitude group (≥1 500 m) and low altitude group (<1 500 m); meanwhile, these patients were divided into Shamblin type I, type II and type III groups according to Shamblin classification criteria; differences in general data and clinical features among patients from different altitude groups or Shamblin subgroups were compared. Independent influencing factors for Shamblin type III CBTs were analyzed by multivariate ordered Logistic regression. Results:(1) A total of 359 patients were enrolled in the study, including 276 females and 83 males, aged (48.80±12.07) years; 211 patients were into the high altitude group and 148 into the low altitude group; 165 patients were into Shamblin type I group, 146 into Shamblin type II group, and 48 into Shamblin type III group. (2) Compared with those in the low altitude group, patients in the high altitude group had higher proportion of females, older age, lower proportion of Han nationality, higher proportion of Shamblin type I, smaller tumor volume, lower platelet count, higher red blood cell count, hematocrit, hemoglobin level, platelet distribution width and mean platelet volume, and higher large platelet percentage, with significant differences ( P<0.05). (3) Compared with those in the Shamblin type I group, patients in the Shamblin type III group had younger age, lower resident altitude, larger tumor volume, longer time interval from onset to diagnosis, higher proportion of unintentional tumor discovery, larger volume of intraoperative blood loss, lower hemoglobin level, hematocrit, mean erythrocyte volume, and mean hemoglobin concentration, decreased erythrocyte distribution width variable coefficient, and increased platelet count, with significant differences ( P<0.05). Compared with those in the Shamblin type II group, patients in Shamblin type III group had younger age, larger tumor volume, longer time interval from onset to diagnosis, larger volume of intraoperative blood loss, lower hemoglobin, hematocrit and mean erythrocyte volume, higher erythrocyte distribution width variable coefficient and platelet count, with significant differences ( P<0.05). (4) Age ( OR=0.960, 95% CI: 0.942-0.977, P<0.001), residence altitude ( OR=0.992, 95% CI: 0.990-0.999, P=0.020) and time interval from onset to diagnosis ( OR=1.009, 95% CI: 1.005-1.014, P<0.001) were independent influencing factors for Shamblin type III CBTs. Conclusions:More females than males are noted in patients with adult CBTs in Northwest China, and more CBTs patients live at high altitude, with Shamblin type I enjoying the highest proportion. More female and old patients lived at high altitude is noted than those lived at low altitude; patients with Shamblin type III have the youngest age, lowest altitude, and longest time interval from onset to diagnosis. CBTs patients with young age, low residence altitude, and long time interval from onset to diagnosis are more likely to develop Shamblin type III.

Objective To investigate the protective effects and underlying mechanisms of sodium butyrate on rats exposed to hypoxia and cold conditions.Methods Fifty-eight male SD rats (aged 7～8 weeks,weighing 240～260 g ) were randomly divided into normoxia normothermia saline control (NNC ) group (n=10),normoxia normothermia sodium butyrate(NNB)group(n=10),hypoxia cold saline control (HCC) group (n=19),and hypoxia cold sodium butyrate (HCB)group (n=19).The intragastric dose of sodium butyrate was 200 mg/kg for the NNB and HCB groups,while the NNC and HCC groups were given normal saline of 5 mL/kg.①After continuous intragastric administration for 7 d,the rats in the HCC and HCB groups were placed in a low-pressure hypoxic chamber to simulate an altitude of 5000 m and exposed to a temperature of 8 ℃ for 7 d.Subsequently,blood samples were collected from the abdominal aorta for blood gas analysis,blood routine test,and detection for serum biochemical indicators.ELISA was used to determine serum inflammatory cytokines and endocrine hormones.The rats in the NNC and NNB groups(n=10)were fed outside the chamber and underwent the same tests in 7 d later to evaluate the protective effects of sodium butyrate on the body.②Core body temperature monitoring was conducted to assess the impact of sodium butyrate on the rmoregulation in rats exposed to hypoxia and cold(n=3).③Hypoxia exercise tolerance of the HCC group and HCB group in a hypoxic chamber (11％O2 )was evaluated for their hypoxia resistance (n=6).Results Compared to the NNC group,the rats in the HCC group exhibited significant decreases in arterial oxygen saturation (SaO2 )and arterial oxygen partial pressure (PaO2 ),serum levels of IL-4,estradiol (E2)and cortisol (F),core temperature,and exercise duration (P<0.05),and had notably increased levels of red blood cell (RBC)count,hemoglobin (HGB),hematocrit (HCT),cardiac troponin (CRDAC-T),uric acid (UA),alanine aminotransferase (ALT),total cholesterol (TC),low-density lipoprotein (LDL),IL-6 and granulocyte-macrophage colony-stimulating factor (GM-CSF)(P<0.05).Compared to the HCC group,the rats in the HCB group exhibited significant increases in SaO2,PaO2,IL-4,E2,F,corticotropin releasing hormone (CRH)and adrenocorticotropic hormone (ACTH)(P<0.05),remarkably longer exercise duration under hypoxic exposure (P<0.05 ),but decreases in RBC count,serum levels of HGB,HCT,CRDAC-T,UA,ALT,TC,LDL,IL-6,GM-CSF and free thyroxine (FT4 ),and core temperature (P<0.05).Conclusion Sodium butyrate exhibits protective effects on rats exposed to hypoxia and cold conditions,and it is helpful in their adaptation to these hypoxia and cold environments.

Objective:To analyze the hemodynamic changes of different types of unruptured intracranial aneurysms before and after flow diverter (FD) treatment with computational fluid dynamics (CFD), and lay research foundation for precision treatment and prognosis evaluation for unruptured intracranial aneurysms.Methods:Four patients with different types of unruptured intracranial aneurysms, admitted to Department of Neurosurgery, First Affiliated Hospital of Air Force Medical University from January 2022 to March 2022, were chosen. Digital subtraction angiography (DSA) data of the patients before and immediately after surgery were collected. Morphological and hemodynamic parameters of the aneurysms were calculated by 3D reconstruction, finite element simulation, and CFD methods: ostium ratio (OsR), neck ratio (NR), area ratio (ArR), volume ratio (VoR), wall shear stress (WSS), normalized wall shell stress (NWSS), blood inflow, relative inflow, aneurysm average velocity, parent artery average velocity, normalized velocity, residual flow volume (RFV), and inflow concentration index (ICI); differences of these indexes before and after treatment were compared.Results:The OsR of 6 aneurysms was 0.225, 0.267, 0.265, 0.389, 1.000, 1.000, respectively; NR was 1.220, 0.274, 1.090, 1.587, 2.809, and 4.019, respectively; ArR was 0.608 and 0.224, 0.623, 3.462, 1.225 and 1.784, respectively; and VoR was 0.386, 0.052, 0.212, 3.462, 0.422 and 1.882, respectively. The parameters of WSS, NWSS, blood inflow, relative inflow, aneurysm average velocity, parent artery average velocity, normalized velocity, RFV, and ICI decreased obviously after FD implantation.Conclusion:On the basis of 3D reconstruction combined with FD/coil virtual implantation, CFD-based hemodynamic analysis can obtain accurate parameters of different types of intracranial aneurysms before and after FD treatment.

Objective:To investigate the influence and mechanism of stromal interaction molecule 1 (STIM1) in microglia/macrophages M1 activation after cerebral ischemia-reperfusion injury.Methods:(1) Animal experiment: 20 male C57BL/6J mice were randomly divided into sham-operated (Sham) group, middle cerebral artery occlusion and reperfusion (MCAO/R) group, MCAO/R+si-Ctrl group, and MCAO/R+si-STIM1 group ( n=5); MCAO/R models were established in mice of the latter 3 groups; empty vector control virus and STIM1 gene knockout lentivirus were transfected into mice in the MCAO/R+si-Ctrl group and MCAO/R+si-STIM1 group. The transfection efficiency of STIM1 and the expression of microglia/macrophages M1 activation marker cluster of differentiation 86 (CD86) in each group were observed. (2) Cell experiment: primary microglia were divided into Ctrl group, oxygen-glucose deprivation/re-oxygenation (OGD/R) group, OGD/R+si-Ctrl group, OGD/R+si-STIM1 group, OGD/R+solvent group, and OGD/R+4-phenylbutyric acid (4-PBA) group; OGD/R models were established in the later 5 groups; empty vector control virus and STIM1 gene knockout lentivirus were transfected into mice in the OGD/R+si-Ctrl group and OGD/R+si-STIM1 group; cells in the OGD/R+4-PBA group were pre-treated with 1 mmol/L 4-PBA for 1 h at 24 h before OGD/R modelling to inhibit endoplasmic reticulum stress (ERS), and cells in the OGD/R+solvent group were pre-treated with 0.5% dimethyl sulfoxide (DMSO) for 1 h at the same time. Reverse transcription quantitative polymerase chain reaction (RT-qPCR), ELISA, Western blotting and other methods were used to detect the levels of CD86, tumour necrosis factor-α ( TNF-α) mRNA, interleukin (IL)-1β, and ERS-related proteins (transcription factor C/EBP homologous protein [CHOP], activated transcription factor 4 [ATF4]) in these cells. Results:(1) Animal experiment: the STIM1 expression in MCAO/R+si-STIM1 group was significantly lower than that in Sham group, MACO/R group and MCAO/R+si-Ctrl group ( P<0.05); as compared with that in the MACO/R group and MCAO/R+si-Ctrl group, the number of microglia/macrophages co-expressing CD86 and Iba-1 around the ischemic foci of mice in the MCAO/R+si-STIM1 group was significantly decreased ( P<0.05). (2) Cell experiment: as compared with those in the OGD/R group and OGD/R+si-Ctrl group, the expression levels of STIM1, CD86, and TNF-α mRNA, and supernatant IL-1β content in the OGD/R+si-STIM1 group were significantly decreased ( P<0.05); as compared with those in the OGD/R group and OGD/R+si-CTRL group, the ATF4 and CHOP expression levels in OGD/R+si-STIM1 group were significantly decreased ( P<0.05); as compared with those in the OGD/R group and OGD/R+solvent group, the CD86 level, TNF-α mRNA expression level and IL-1β content in the OGD/R+4-PBA group were significantly decreased ( P<0.05). Conclusion:STIM1 affects microglia/macrophages M1 activation after ischemia-reperfusion injury by regulating ERS level.

Morchella is a genus of fungi with the ability to concentrate Cd both in the fruit-body and mycelium. However, the molecular mechanisms conferring resistance to Cd stress in Morchella are unknown. Here, RNA-based transcriptomic sequencing was used to identify the genes and pathways involved in Cd tolerance in Morchella spongiola. 7444 differentially expressed genes (DEGs) were identified by cultivating M. spongiola in media containing 0.15, 0.90, or 1.50 mg/L Cd2+ . The DEGs were divided into six sub-clusters based on their global expression profiles. GO enrichment analysis indicated that numerous DEGs were associated with catalytic activity, cell cycle control, and the ribosome. KEGG enrichment analysis showed that the main pathways under Cd stress were MAPK signaling, oxidative phosphorylation, pyruvate metabolism, and propanoate metabolism. In addition, several DEGs encoding ion transporters, enzymaticon-enzymatic antioxidants, and transcription factors were identified. Based on these results, a preliminary gene regulatory network was firstly proposed to illustrate the molecular mechanisms of Cd detoxification in M. spongiola. These results provide valuable insights into the Cd tolerance mechanism of M. spongiola and constitute a robust foundation for further studies on detoxification mechanisms in macrofungi that could potentially lead to the development of new and improved fungal bioremediation strategies.

Severe aplastic anemia II (SAA-II) progresses from non-severe aplastic anemia (NSAA). The unavailability of efficacious treatment has prompted the need for haploidentical bone marrow transplantation (haplo-BMT) in patients lacking a human leukocyte antigen (HLA)-matched donor. This study aimed to investigate the efficacy of haplo-BMT for patients with SAA-II. Twenty-two patients were included and followed up, and FLU/BU/CY/ATG was used as conditioning regimen. Among these patients, 21 were successfully engrafted, 19 of whom survived after haplo-BMT. Four patients experienced grade II-IV aGvHD, including two with grade III-IV aGvHD. Six patients experienced chronic GvHD, among whom four were mild and two were moderate. Twelve patients experienced infections during BMT. One was diagnosed with post-transplant lymphoproliferative disorder and one with probable EBV disease, and both recovered after rituximab infusion. Haplo-BMT achieved 3-year overall survival and disease-free survival rate of 86.4% ± 0.73% after a median follow-up of 42 months, indicating its effectiveness as a salvage therapy. These promising outcomes may support haplo-BMT as an alternative treatment strategy for patients with SAA-II lacking HLA-matched donors.
Anemia, Aplastic/therapy* ; Bone Marrow Transplantation ; Graft vs Host Disease ; HLA Antigens ; Hematopoietic Stem Cell Transplantation ; Humans ; Transplantation Conditioning

Morchella is a genus of fungi with the ability to concentrate Cd both in the fruit-body and mycelium. However, the molecular mechanisms conferring resistance to Cd stress in Morchella are unknown. Here, RNA-based transcriptomic sequencing was used to identify the genes and pathways involved in Cd tolerance in Morchella spongiola. 7444 differentially expressed genes (DEGs) were identified by cultivating M. spongiola in media containing 0.15, 0.90, or 1.50 mg/L Cd2+ . The DEGs were divided into six sub-clusters based on their global expression profiles. GO enrichment analysis indicated that numerous DEGs were associated with catalytic activity, cell cycle control, and the ribosome. KEGG enrichment analysis showed that the main pathways under Cd stress were MAPK signaling, oxidative phosphorylation, pyruvate metabolism, and propanoate metabolism. In addition, several DEGs encoding ion transporters, enzymaticon-enzymatic antioxidants, and transcription factors were identified. Based on these results, a preliminary gene regulatory network was firstly proposed to illustrate the molecular mechanisms of Cd detoxification in M. spongiola. These results provide valuable insights into the Cd tolerance mechanism of M. spongiola and constitute a robust foundation for further studies on detoxification mechanisms in macrofungi that could potentially lead to the development of new and improved fungal bioremediation strategies.