OBJECTIVE: To investigate the short-term efficacy and safety of low-dose venetoclax combined with CHG (cytarabine+homoharringtonine+G-CSF) priming regimen in patients with acute myeloid leukemia (AML) and high-risk myelodysplastic syndrome (MDS) ineligible for intensive chemotherapy. METHODS: The data of 14 patients with AML or high-risk MDS admitted to the department of hematology/oncology of the First Hospital of Tsinghua University and 2 cooperative institutions from July 2022 to August 2023 were retrospectively analyzed. All the patients were treated with low-dose venetoclax combined with CHG priming regimen and the early induction (one course) efficacy and adverse reactions were observed. RESULTS: Among the 14 patients, 10 were males and 4 were females, with a median age of 69.5 (46-83) years. After 1 cycle of induction chemotherapy, the complete remission (CR) rate was 64.3% (9/14) and overall response rate (ORR) was 78.6% (11/14). Among the 10 patients with adverse prognosis according to cytogenetics and molecular genetics, the CR rate was 50.0% (5/10), and ORR was 70.0% (7/10). In 7 patients with TP53 mutation, the CR rate was 42.9% (3/7) and ORR was 71.4% (5/7). In the 6 patients with complex karyotype, CR rate was 33.3% (2/6) and ORR was 66.7% (4/6). While the CR rate and ORR of 8 non-complex karyotype patients were both 87.5% (7/8), and the difference in CR rate between patients with complex karyotype and non-complex karyotype was statistically significant ( P < 0.05). The adverse reactions of chemotherapy were tolerable, without early treatment-related deaths. CONCLUSION Low-dose venetoclax combined with CHG priming regimen can be used as an effective treatment for AML and high-risk MDS patients who are ineligible for intensive chemotherapy, and it is safe and worthy of clinical application.
Humans ; Leukemia, Myeloid, Acute/drug therapy* ; Aged ; Male ; Female ; Sulfonamides/therapeutic use* ; Middle Aged ; Myelodysplastic Syndromes/drug therapy* ; Bridged Bicyclo Compounds, Heterocyclic/therapeutic use* ; Aged, 80 and over ; Retrospective Studies ; Cytarabine/administration & dosage* ; Antineoplastic Combined Chemotherapy Protocols/therapeutic use* ; Homoharringtonine/therapeutic use*

Objective To explore the role of miR-1260b targeting activating transcription factor 6β(ATF6β)in osteogenic differentia-tion of human periodontal ligament stem cells(hPDLSCs).Methods The periodontal ligament tissue was scraped from the third molar extracted from the patient with periodontitis,and hPDLSCs were isolated and cultured.The proportions of positive cells of CD34,CD45,CD29,CD90 and CD105 were detected by flow cytometry,and the expression of vimentin and pan-cytokeratin were detected by immunofluo-rescence staining.The cellular luciferase activities of ATF6β-WT and ATF6β-MUT in the miR-1260b mimic+WT/MUT group(transfected with miR-1260b mimic and ATF6β-WT/MUT)and the mimic-NC group(transfected with miR-NC and ATF6β-WT/MUT)were analyzed by the dual luciferase reporter gene assay.In addition,the cells were divided into the miR-1260b mimic group(transfected with miR-1260b mimic alone),the ATF6β-OE group(transfected with the ATF6β overexpression vector alone),the combined group(transfected with miR-1260b mimic and ATF6β overexpression vector simultaneously),and the NC group(transfected with empty vector).Alizarin red S staining and alkaline phosphatase staining were performed on hPDLSCs in the NC group,the miR-1260b mimic group,the ATF6β-OE group and the combined group.Meanwhile,the mRNA expression levels of miR-1260b,ATF6β,Runt-related transcription factor 2(Runx2),osteocalcin(OCN),and osteopontin(OPN)in each group of cells were detected by qRT-PCR.The protein expressions of glucose-regulated protein 78(GRP78),C/EBP homologous protein(CHOP),and protein kinase R-like endoplasmic reticulum kinase(PERK)in each group of cells were detected by Western blot.Results The flow cytometry detection showed that CD29,CD90 and CD105 on the surface of the isolated and cultured cells were positive,while CD34 and CD45 were negative.Immunofluorescence staining showed that vimentin in the cells was positive and pan-cytokeratin was negative,which was in line with the characteristics of hPDLSCs.Luciferase assay showed that miR-1260b mimic could significantly reduce the luciferase activity of ATF6β-WT(P<0.05),but had no significant effect on the luciferase activity of ATF6β-MUT(P>0.05).After alizarin red S staining,the staining of hPDLSCs gradually deepened after 3 days,5 days and 7 days of culture,suggesting an enhanced osteogenic differentiation ability.With the enhancement of osteogenic differentiation ability of hPDLSCs,the expression of miR-1260b in the cells showed a gradually increasing trend,while the mRNA and protein expression levels of ATF6β showed a gradually decreasing trend(P<0.05).The results of alizarin red S staining and alkaline phosphatase staining showed that the proportions of positive area in the miR-1260b mimic group were higher than those in the NC group(P<0.05),while the proportions of positive area in the ATF6β-OE group and the combined group were lower than those in the NC group(P<0.05).The mRNA and protein levels of ATF6β in the miR-1260b mimic group were lower than those in the NC group(P<0.05),while the mRNA and protein levels of ATF6β in the ATF6β-OE group and the combined group were higher than those in the NC group(P<0.05).The mRNA levels of Runx2,OCN and OPN in the miR-1260b mimic group were significantly higher than those in the NC group(P<0.05).The mRNA levels of Runx2,OCN and OPN in the ATF6β-OE group and the combined group were significantly lower than those in the NC group(P<0.05).The protein expression levels of GRP78,CHOP and PERK in the miR-1260b mimic group were significantly lower than those in the NC group(P<0.05).The protein expression levels of GRP78,CHOP and PERK in the ATF6β-OE group and the combined group were significantly higher than those in the NC group(P<0.05).Conclusion miR-1260b can inhibit endoplasmic reticulum stress level and promote osteogenic differentiation of hPDLSCs by targeting ATF6β.

Objective:To investigate the significance of serum β2-microglobulin(β2-MG)for survival and relapse of patients with diffuse large B-cell lymphoma(DLBCL)in the rituximab era.Methods:Clinical data of 92 patients with DLBCL admitted from December 2003 to July 2015 were retrospectively analyzed.The optimal cutoff value of β2-MG levels for predicting prognosis of the DLBCL patients was determined using receiver operating characteristic(ROC)curve.Kaplan-Meier analysis was used to estimate progression-free survival(PFS)and overall survival(OS).Cox logistic regression analysis was used to explore potential prognostic factors associated with survival.Binary logistic regression analysis was used to analyze the relationship between various factors and relapse.Results:The most discriminative cutoff value for β2-MG level was determined to be 2.25 mg/L by the ROC curve.Subgroup analysis showed that patients in the elevated β2-MG(＞2.25 mg/L)group had significantly worse PFS(P=0.006)and a trend toward worse OS compared with those in the low β2-MG(≤2.25 mg/L)group(P=0.053).Univariate analysis showed that elevated β2-MG,age＞60 years,Ann Arbor stage Ⅲ-Ⅳ,as well as IPI score ≥ 3 were associated with worse PFS.Binary logistic regression analysis showed that age＞60 years and β2-M G＞2.25 mg/L were potential influencing factors for relapse of DLBCL patients.Conclusion:Serum β2-MG might be an important predictor for the survival and relapse of DLBCL patients in the rituximab era.

Objective To explore the role of miR-1260b targeting activating transcription factor 6β(ATF6β)in osteogenic differentia-tion of human periodontal ligament stem cells(hPDLSCs).Methods The periodontal ligament tissue was scraped from the third molar extracted from the patient with periodontitis,and hPDLSCs were isolated and cultured.The proportions of positive cells of CD34,CD45,CD29,CD90 and CD105 were detected by flow cytometry,and the expression of vimentin and pan-cytokeratin were detected by immunofluo-rescence staining.The cellular luciferase activities of ATF6β-WT and ATF6β-MUT in the miR-1260b mimic+WT/MUT group(transfected with miR-1260b mimic and ATF6β-WT/MUT)and the mimic-NC group(transfected with miR-NC and ATF6β-WT/MUT)were analyzed by the dual luciferase reporter gene assay.In addition,the cells were divided into the miR-1260b mimic group(transfected with miR-1260b mimic alone),the ATF6β-OE group(transfected with the ATF6β overexpression vector alone),the combined group(transfected with miR-1260b mimic and ATF6β overexpression vector simultaneously),and the NC group(transfected with empty vector).Alizarin red S staining and alkaline phosphatase staining were performed on hPDLSCs in the NC group,the miR-1260b mimic group,the ATF6β-OE group and the combined group.Meanwhile,the mRNA expression levels of miR-1260b,ATF6β,Runt-related transcription factor 2(Runx2),osteocalcin(OCN),and osteopontin(OPN)in each group of cells were detected by qRT-PCR.The protein expressions of glucose-regulated protein 78(GRP78),C/EBP homologous protein(CHOP),and protein kinase R-like endoplasmic reticulum kinase(PERK)in each group of cells were detected by Western blot.Results The flow cytometry detection showed that CD29,CD90 and CD105 on the surface of the isolated and cultured cells were positive,while CD34 and CD45 were negative.Immunofluorescence staining showed that vimentin in the cells was positive and pan-cytokeratin was negative,which was in line with the characteristics of hPDLSCs.Luciferase assay showed that miR-1260b mimic could significantly reduce the luciferase activity of ATF6β-WT(P<0.05),but had no significant effect on the luciferase activity of ATF6β-MUT(P>0.05).After alizarin red S staining,the staining of hPDLSCs gradually deepened after 3 days,5 days and 7 days of culture,suggesting an enhanced osteogenic differentiation ability.With the enhancement of osteogenic differentiation ability of hPDLSCs,the expression of miR-1260b in the cells showed a gradually increasing trend,while the mRNA and protein expression levels of ATF6β showed a gradually decreasing trend(P<0.05).The results of alizarin red S staining and alkaline phosphatase staining showed that the proportions of positive area in the miR-1260b mimic group were higher than those in the NC group(P<0.05),while the proportions of positive area in the ATF6β-OE group and the combined group were lower than those in the NC group(P<0.05).The mRNA and protein levels of ATF6β in the miR-1260b mimic group were lower than those in the NC group(P<0.05),while the mRNA and protein levels of ATF6β in the ATF6β-OE group and the combined group were higher than those in the NC group(P<0.05).The mRNA levels of Runx2,OCN and OPN in the miR-1260b mimic group were significantly higher than those in the NC group(P<0.05).The mRNA levels of Runx2,OCN and OPN in the ATF6β-OE group and the combined group were significantly lower than those in the NC group(P<0.05).The protein expression levels of GRP78,CHOP and PERK in the miR-1260b mimic group were significantly lower than those in the NC group(P<0.05).The protein expression levels of GRP78,CHOP and PERK in the ATF6β-OE group and the combined group were significantly higher than those in the NC group(P<0.05).Conclusion miR-1260b can inhibit endoplasmic reticulum stress level and promote osteogenic differentiation of hPDLSCs by targeting ATF6β.

Aim To study the role and mechanism of ML210 in glioma.Methods The cell viability was detected by CCK8 assay.The percentage of dead cells was detected by SYTOXstaining.The role of ferroptosis-signaling pathway in gliomas was detected bygenomics.Cell proliferation was observed by EdU staining and clone formation assay.Cell migration ability was detec-ted by scratch healing assay.The apoptosis was detec-ted by flow cytometry.Cell mitochondrial function was assesses by JC-1 staining.The mechanism of action of ML210 was detected by molecular docking coupled with immunoblotting assay(Western blot).The levels of ROS,MDA were observed by ELISA.Results Compared with the control group,ML210 treatment dose-dependently decreased glioma cell viability,in-hibited cell proliferation,migration,and increased cell apoptosis and mitochondrial dysfunction,which were reversed by ferroptosis antagonists.Gene microarray screening showed that 688 genes of the ferroptosissig-naling pathway were aberrant and 10 signaling path-ways were altered in gliomas.Molecular docking re-sults showed that ML210 binding to GPX4 significantly inhibited the protein expression level of GPX4 and pro-moted the elevation of ROS and MDA levels.Conclu-sions ML210 produces anti-glioma cells via GPX4-mediated ferroptosis pathway.

Aim To study the role and mechanism of ML210 in glioma.Methods The cell viability was detected by CCK8 assay.The percentage of dead cells was detected by SYTOXstaining.The role of ferroptosis-signaling pathway in gliomas was detected bygenomics.Cell proliferation was observed by EdU staining and clone formation assay.Cell migration ability was detec-ted by scratch healing assay.The apoptosis was detec-ted by flow cytometry.Cell mitochondrial function was assesses by JC-1 staining.The mechanism of action of ML210 was detected by molecular docking coupled with immunoblotting assay(Western blot).The levels of ROS,MDA were observed by ELISA.Results Compared with the control group,ML210 treatment dose-dependently decreased glioma cell viability,in-hibited cell proliferation,migration,and increased cell apoptosis and mitochondrial dysfunction,which were reversed by ferroptosis antagonists.Gene microarray screening showed that 688 genes of the ferroptosissig-naling pathway were aberrant and 10 signaling path-ways were altered in gliomas.Molecular docking re-sults showed that ML210 binding to GPX4 significantly inhibited the protein expression level of GPX4 and pro-moted the elevation of ROS and MDA levels.Conclu-sions ML210 produces anti-glioma cells via GPX4-mediated ferroptosis pathway.

Objective To study the protective effect of Irisin in doxorubicin(Dox)induced-Cardiomyopathy and its possible mechanism.Methods AC 16 cells were used to construct Dox injury model and divided into control group(AC 16 cells were cultured with complete medium),Irisin group(AC16 cells were treated with 10 ng·L-1 Irisin for 24 h),Dox group(AC 16 cells were treated with 4 μmol·L-1 Dox for 24 h),Dox+Irisin group(AC 16 cells were pretreated with 10 ng·L-1 Irisin for 2 h,and then treated with 4 pmol·L-1 Dox for 24 h).Cell counting kit-8(CCK-8),terminal deoxynucleotidyl transferase-mediated nick end labeling(TUNEL)and lactate dehydrogenase(LDH)were used to detect the proliferation,apoptosis and mortality of AC 16 cells.Western blot was used to detect the expression levels of nuclear factor-κB(NF-κB)signaling pathway and apoptotic factors B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax)and caspase-9 protein.Mito-Tracker Red CMXRos probe was used to detect mitochondrial membrane potential.Results In the contrl group,Irisin group,Dox group,Dox+Irisin group,the rate of apoptosis were(0.97±0.09)％,0,(42.80±6.70)％,(11.74±1.79)％;the expression of Bax protein were 0.85±0.01,0.36±0.02,1.15±0.07,0.37±0.11;the expression of caspase-9 protein were 0.52±0.02,0.59±0.03,1.11±0.02,0.67±0.08;the expression of Bcl-2 protein were 1.01±0.04,1.05±0.25,0.43±0.02 and 0.99±0.30;the probability of mitochondrial damage were(0.02±0.01)％,(0.5±0.15)％,(38.6±2.39)％,(1.58±0.54)％.The difference of the above indexes between the contrl group and the Dox group were statistically significant(all P＜0.05);the difference between Dox group and Dox+Irisin group were statisically significant(all P＜0.05).Conclusion Irisin could reduce the expression level of Bax,caspase-9,p-NF-κB,and p-mTOR caused by Dox,increase the expression level of Bcl-2,ameliorate the myocardial damage caused by Dox,and reduce cardiotoxicity.

Objective To investigate the incidence rate,clinical characteristics,and prognosis of neonatal stroke in Shenzhen,China.Methods Led by Shenzhen Children's Hospital,the Shenzhen Neonatal Data Collaboration Network organized 21 institutions to collect 36 cases of neonatal stroke from January 2020 to December 2022.The incidence,clinical characteristics,treatment,and prognosis of neonatal stroke in Shenzhen were analyzed.Results The incidence rate of neonatal stroke in 21 hospitals from 2020 to 2022 was 1/15 137,1/6 060,and 1/7 704,respectively.Ischemic stroke accounted for 75％(27/36);boys accounted for 64％(23/36).Among the 36 neonates,31(86％)had disease onset within 3 days after birth,and 19(53％)had convulsion as the initial presentation.Cerebral MRI showed that 22 neonates(61％)had left cerebral infarction and 13(36％)had basal ganglia infarction.Magnetic resonance angiography was performed for 12 neonates,among whom 9(75％)had involvement of the middle cerebral artery.Electroencephalography was performed for 29 neonates,with sharp waves in 21 neonates(72％)and seizures in 10 neonates(34％).Symptomatic/supportive treatment varied across different hospitals.Neonatal Behavioral Neurological Assessment was performed for 12 neonates(33％,12/36),with a mean score of(32±4)points.The prognosis of 27 neonates was followed up to around 12 months of age,with 44％(12/27)of the neonates having a good prognosis.Conclusions Ischemic stroke is the main type of neonatal stroke,often with convulsions as the initial presentation,involvement of the middle cerebral artery,sharp waves on electroencephalography,and a relatively low neurodevelopment score.Symptomatic/supportive treatment is the main treatment method,and some neonates tend to have a poor prognosis.

Objective:To investigate which indicator is more advantageous when using arterial oxygen saturation(SaO2)and fingertip pulse oxygen saturation(SpO2)for blood oxygen detection in patients with hyperleukocytic acute leukemia(HAL).Methods:In this prospective research,the difference between SaO2 and SpO2 of 18 HAL patients(observation group)and 14 patients(control group),as well as the relationship between the difference and white blood cell(WBC)counts were analyzed.Results:SaO2 was lower than SpO2 in the observation group(P＜0.05),and SpO2-SaO2 difference was positively correlated with WBC counts(r=0.47).However,there was no statistical difference between SaO2 and SpO2 in the control group.SaO2 and PO2 showed a downward trend with the prolongation of detection time after arterial blood was collected in the observation group,but there was no statistical difference.There was no downward trend of SaO2 and PO2 in the control group.Conclusion:HAL patients have a phenomenon where SaO2 is lower than SpO2,that is pseudohypoxemia,and this phenomenon may be caused by excessive consumption of oxygen by the leukemia cells in vitro.SpO2 can be monitored bedside in real time and is non-invasive,it is a better way to detect the blood oxygen status of HAL patients.

Objective:To investigate the effect of feeder layer cells expressing interleukin(IL)-21 on the amplification of NK cells in vitro.Methods:The K562 cell line with IL-21 expression on its membrane was constructed by electroporation,and co-cultured with NK cells after inactivation.The proliferation of NK cells was observed.The killing function of the amplified NK cells in vitro was evaluated by the lactate dehydrogenase(LDH)and interferon-γ(IFN-y)release assay.A colorectal cancer xenograft model in NOD/SCID mice was established,and a blank control group,a NK cell group and an amplified NK cell group were set up to detect the tumor killing effect of amplified NK cells in vivo.Results:K562 cells expressing IL-21 on the membrane were successfully constructed by electroporation.After co-culturing with K562 cells expressing IL-21 on the membrane for 17 days,the NK cells increased to 700 times,which showed an enhanced amplification ability compared with control group(P＜0.001).In the tumor cell killing experiment in vitro,there was no significant difference in the killing activity on tumor cells between NK cells and amplified NK cells,and there was also no significant difference in mice in vivo.Conclusion:K562 cells expressing IL-21 on the membrane can significantly increase the amplification ability of NK cells in vitro,but do not affect the killing function of NK cells in vitro and in vivo.It can be used for the subsequent large-scale production of NK cells in vitro.