BACKGROUND: Endothelial colony-forming cells (ECFCs) transplantation exerts beneficial impact on angiogenesis of impaired tissues caused by ischemia. However, whether this impact is related to the paracrine effect of ECFCs needs to be studied further.OBJECTIVE: To detect the cytokine secretion profile of human umbilical cord blood derived-ECFCs conditioned medium (ECFCs-CM) and explore the effect of ECFCs-CM on the proliferation, migration and tube formation ability of human umbilical vein endothelial cells (HUVECs).METHODS: Human cord blood derived-ECFCs were isolated, cultured in vitro, and then identified based on the previous studies. The cytokines in serum free ECFCs-CM were detected using a cytokines antibody array. HUVECs were cultured with ECFCs-CM, serum free EBM-2 as control. The proliferation, migration and tube formation abilities of HUVECs were examined by cell counting kit-8, scratch test and Matrigel assay, respectively.RESULTS AND CONCLUSION: The cultured cells demonstrated typical characteristics of ECFCs, which showed cobblestone appearance and were positive for CD34, KDR and CD144, but not CD45 or CD133, uptook Dil-acLDL and bond FITC-UEA-I, and tube-like structures were formed on Matrigel. The cytokine antibody array showed that ECFCs-CM significantly upregulated the expressions of 30 kinds of angiogenic factors. Compared with the control group,the HUVECs cultured with ECFCs-CM showed significantly improved proliferation ability at 24, 48 and 72 hours (P <0.01). The migration rate of HUVECs in the experimental group was significantly higher than that in the control group at 12 and 24 hours (P < 0.01). There were more tubular structures in the experimental group than those in the control group (P < 0.05). These results indicate that ECFCs can promote the bioactivity of mature endothelial cells through paracrine action.

Objective To examine the effect of endothelial colony-forming cells conditioned medium(ECFCs-CM)on biological function of human dermal fibroblasts(HDFs). Methods Human cord blood derived-ECFCs were isolated and identified based on the previous studies. The cytokines in ECFCs-CM were detected using a cytokines antibody array. HDFs were cultured with ECFCs-CM,using serum free EBM-2 as control. The proliferation of HDFs was examined by Cell Counting Kit-8(CCK-8)and the migration was assessed by scratch test assay. The apoptosis of HDFs was detected by flow cytometry. Results The cells isolated from human cord blood demonstrated typical characteristics of ECFCs. The cytokines antibody array indicated that ECFCs-CM contained large amounts of secreted cytokines such as PDGF-BBand EGF. Compared with the control group,the HDFs cultured with ECFCs-CM showed improved proliferation and migration ability. The number of apoptotic cells was smaller than that of the control group under the environment of serum starvation. Conclusion ECFCs-CM can promote the proliferation and migration of HDFs and inhibit the apoptosis of HDFs under the environment of serum starvation.

Objective To explore clinical application of retrograde medial and lateral gastrocnemius muscle flap for soft tissue defects of the middle and lower third of the leg. Methods From August 2008 to December 2009, in our hospital we adopted retrograde medial and lateral gastrocnemius muscle flap to renovate 5 cases of refractory soft tissue defects of the middle and lower third of the leg. Results Five cases of retrograde medial gastrocnemius muscle flap were survived, morphology and function of soft tissue defects were renovated well. Conclusion This operation is an effective and reliable technique for soft tissue defects of the middle and lower third of the leg, which is performed without sacrificing the major blood vessels, probing vascular pedicle and matching vascular anastomosis.

Objective To introduce clinical application of retrograde medial pedicled skin flap of the leg to repair the plantar cutaneous deficiency. Methods From January 2002 to May 2008, in our hospital we adopted retrograde medial pedicled skin flap to renovate 12 cases of plantar cutaneous deficiency. The size of the skin flaps ranged from 10 cm× 7 cm to 13 cm × 12 cm. Results Ten cases of retrograde medial pedicled skin flap were survived, the rest had partial necrosis because of distal blood flow obstacle. One was cured by changing dressings; the other was repaired by secondary operation. Morphology and function of soft tissue defects were renovated well with 6-18 months follow-up. Conclusion This operation is an effective and reliable technique for plantar cutaneous deficiency.

Objective:To study the clinical characteristics of odontogenic cutaneous fistula.Methods:6 cases of odontogenic cutane-ous fistula were represented and the reports of 60 cases of the lesion were reviewed.Results:The fistula was mainly located in cheek, chin,para-nasal part,sub-mandibular area and the lower border of the mandible,and respectively corresponding to mandibular third molar,mandibular incisor,maxillary canine,mandibular first and second molar,the corresponding teeth were mainly in mandible (71 .2%).In middle and old aged patients the lesion usually in para-nasal or mandible area,In younger patients the lesion mostly loca-ted in cheek or sub-mandibular area.After root canal therapy for the teeth with apical periodontitis or extraction of none-curable teeth,the odontogenic cutaneous sinus tract disappeared.Conclusion:Proper treatment of focal teeth can cure the odontogenic cutaneous fistulas.

Objective:To study the effects of miR-20a on the osteogenic differentiation potential of inflammatory periodontal liga-ment cells(IPDLSCs).Methods:Cells were isolated and cultured from the healthy and inflammatory periodontal ligament samples (HPDLSCs and IPDLSCs)respectively.miR-20a expression was analyzed by qRT-PCR.Alizarin red staining,Western blot and PCR were used to evaluate the osteogenic differentiation potential of IPDLSCs after transient transinfection of miR-20a mimics or inhib-itor.Results:miR-20a expression in IPDLSCs was lower than that in HPDLSCs,and the osteogenic differentiation potential of IP-DLSCs were promoted by miR-20a mimics,and reduced by miR-20a inhibitor.Conclusion:The miR-20a in IPDLSCs was down reg-ulated.miR-20a can promote the osteogenic differentiation potential of IPDLSCs.

BACKGROUND:Understanding the difference of miRNA-34s expression in normal tissue and tumor tissue wil contribute to screen out a miRNA with high sensitivity as the specific tumor molecular marker.
OBJECTIVE:To investigate the differential expression of miRNA-34s (miR-34a/b/c) between normal skin and keloid tissue using real-time fluorescent quantitative PCR, and to evaluate the role and mechanisms of miRNA-34s in keloid formation and development.
METHODS:Ten cases of keloid tissue and two cases of normal skin tissue were col ected as specimens. Total RNAs were extracted from keloid and nomal skin tissue by Trizol method, and miRNA-34s were further isolated by Ambion’s miRNA Isolation Kit. Real-time fluorescent quantitative PCR was applied to verify expression levels of microRNA-34s (miR-34a/b/c) in keloid tissue and normal skin tissue.
RESULTS AND CONCLUSION:miRNA-34s (miRNA-34a/b/c) expression was down-regulated in keloid tissue compared with normal skin tissue (P<0.01). The findings showed that miRNA-34s (miRNA-34a/b/c) are involved in keloid formation and development, and down-regulation of the family member may result in neoplastic growth of keloid.

Objective:To investigate the expression of SHIP1 in the patients with acute myeloid leukemia and its effect on the apoptosis of human leukemia cells.Methods:The expression of SHIP1 in the bone marrow of patients with acute myeloid leukemia was detected by Westem blot.U937 cells was transfected with SHIP1 expression vector (pEGFP-SHIP1 group) and empty vector control (pEGFP group) respectively,U937 cells without transfection were used as the control group.Flow cytometry was used to detect the apoptosis of the cells,the expression of SHIP1,Bcl-2,Bax,Akt,p-Akt were detected by western blot.Results:The expression of SHIP1 in the bone marrow of patients with acute myeloid leukemia was significantly lower than that of the normal human bone marrow SHIP 1 (P＜0.01).The SHIP1 and Bax expressions as well as the apoptotic rate ofpEGFP-SHIP1 group were significantly higher than those of the control group(P＜0.01),while the Bcl-2 and p-Akt expressions were significantly lower than those in the control group(P＜0.01).Conclusions:SH-P1 expression was down regulated in the bone marrow of patients with acute myeloid leukemia.SHIP1 could promote the apoptosis of human leukemia cells via Akt signaling pathway.

With the research progress of pathogenesis of JAK gene in myeloproliferative neoplasms (MPN), more tyrosine kinase inhibitors were developed. MPN quantify scoring system is used to determine the efficacy of tyrosine kinase inhibitors for MPN. The choice of tyrosine kinase inhibitors, tyrosine kinase for the relief of MPN symptom burden, etc, become the topics of the 57th American Society of Hematology (ASH) annual meeting.

Objective:To analyze the correlation between clinical features and prognosis or prognostic risk factors in patients with KMT2A::AFF1 gene positive B-ALL.Methods:Retrospective cohort study was conducted. 167 cases of B-ALL admitted to the Shanghai General Hospital and the Naval Medical University Affiliated First Hospital from April 1, 2011 to July 31, 2022 were divided into groups according to gene types. 22 cases with KMT2A::AFF1 positive B-ALL were enrolled as the experimental group, 54 cases with BCR::ABL gene positive B-ALL as control group 1 and 91 cases with KMT2A::AFF1 and BCR::ABL gene negative B-All as control group 2. The median age of first diagnosis in the experimental group, control group 1 and control group 2 were 43.5(30.5, 56), 43.5(34, 55) and 32(24, 46) respectively. The median white blood cell counts of the three groups were 142.4(25.7, 247.2)×10 9/L, 37.6(15.7, 102.2)×10 9/L and 13.4(4.3, 33.0)×10 9/L, respectively. Allo-HSCT rates in three groups were 45.5%, 72.2% and 72.5% respectively. Using SPSS 26.0 software, the statistical methods of nonparametric rank sum test, chi-square test, Kaplan-Meier and Cox regression were used to analyze and compare the differences in clinical characteristics, chemotherapy and prognosis between the experimental group and the control groups, and to analyze the risk factors and the differences in prognosis of allo-HSCT in the experimental group. Results:The age difference between the experimental group and the control group 2 was significant ( Z=-2.151, P=0.031). The white blood cell count in experimental group was significantly higher than that in control group 1 ( Z=-2.363, P=0.018) and control group 2 ( Z=-4.886, P<0.001). The rate of allo-HSCT in experimental group was lower than that in control group 1(45.5% vs 72.2%, χ 2=4.890, P=0.027) and control group 2 (45.5% vs 72.5%, χ 2=5.897, P=0.015). The remission rates of the patients in three groups after receiving one course of chemotherapy were 60%(12/20), 83.3%(45/54) and 76.6%(69/90); the remission rates after two courses of chemotherapy were 25%(5/20), 7.4%(4/54) and 12.2% (11/90), and the non-remission rates of more than two courses of treatment were 15%(3/20), 9.3%(5/54) and 11.1%(10/90), respectively. The effect of chemotherapy in experimental group was worse than that in control group 1 ( Z=-1.979, P=0.048). There was no significant difference between the three groups in sex, whether the chromosome is a standard-risk karyotype, hemoglobin at the time of initial onset, platelet count and percentage of bone marrow blast cells. The overall survival rate (OS) of experimental group was significantly lower than that of control group 1 and control group 2(23.9% vs 36.7%, χ 2=7.608, P=0.006 and 23.9% vs. 44.8%, χ 2=6.442, P=0.011), and the 3-year recurrence-free survival (RFS) was also lower than that of the other two groups (14.0% vs 57.6%, χ 2=17.823, P<0.001 and 14.0% vs 48.2%, χ 2=16.432, P<0.001). There was a significant difference in the total OS rate between the experimental group and the group without allo-HSCT (45.0% vs 9.2%, χ 2=15.254, P<0.001). Univariate analysis showed that age was the risk factor of RFS in the experimental group, and allo-HSCT therapy was the protective factor of OS. Multivariate analysis showed that allo-HSCT was an independent protective factor for OS in the experimental group. Conclusions:Patients with KMT2A::AFF1 positive B-ALL had higher white blood cells, less sensitivity to chemotherapy and poor prognosis. Age was a risk factor of RFS in KMT2A::AFF1 positive B-ALL, and allo-HSCT could improve the prognosis.