Objective:To explore the genetic etiology for a child with Progressive external ophthalmoplegia with mitochondrial DNA deletions, autosomal dominant 6 (PEOA6).Methods:A child who had attended the Women and Children′s Hospital Affiliated to Ningbo University on 7 August, 2023 was selected as the study subject. Clinical data of the child were analyzed retrospectively. The child and her parents were subjected to whole exome sequencing (WES), and candidate variant was verified by Sanger sequencing and bioinformatic analysis. This study was approved by Medical Ethics Committee of the Women and Children′s Hospital Affiliated to Ningbo University (Ethics No. EC2020-048).Results:The child, a 7-year-old female, had presented with limb muscle pain, amyosthenia, significantly increased creatine kinase, congenital diaphragmatic hernia and recurrent respiratory tract infections. WES revealed that she has harbored a heterozygous c. 1590G>C (p.L530F) variant of the DNA2 gene, which was verified to have a de novo origin by Sanger sequencing. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the c.1590G>C was rated as a likely pathogenic variant (PS2+ PM2_Supporting+ PP3). Conclusion:The c.1590G>C (p.L530F) variant of the DNA2 gene probably underlay the PEOA6 in this child.

Objective:Explore the feasibility and advantages of using proximal and distal ends of thoracodosal artery and vein as recipient vessels in deep inferior epigastric perforator flap immediate breast reconstruction.Methods:The clinical data of patients who underwent breast reconstruction surgery using the proximal and distal ends of the thoracodorsal vein as recipient vessels at the Department of Breast and Thyroid Surgery of the Second Affiliated Hospital of Hainan Medical University from March 2022 to June 2023 were analyzed retrospectively. Preoperative examinations included thoracoabdominal angiography and color Doppler ultrasonic localization of the main trunk and perforators of the inferior epigastric vessels. The procedure began with mastectomy and axillary lymph node dissection, followed by the isolation of bilateral perforators and the main trunk of the abdominal flap. The main trunks of the bilateral inferior epigastric arteries were then transected, and their vascular pedicles exposed and anastomosed respectively to the proximal and distal ends of the thoracodorsal artery and vein. Both arteries and veins were joined end-to-end. The flap after trimming and reconstruction was then implanted into the cavity left after mastectomy through the incision. Breast positioning was performed with the patient in a knee-bent and hip-flexed position. After adjusting the shape of the reconstructed breast. The donor site was closed, the umbilicus was reconstructed, drainage tubes were placed, and the breast incision was closed. Postoperative follow-up monitored complications associated with the flap and patient satisfaction with the breast reconstruction, utilizing a self-assessment method.Results:Four female patients were included, aged (46.0±6.5) years, ranging from 37 to 52 years. All four patients had bilateral vascular pedicles in the donor area, with three patients having thoracodorsal vessels at the distal and proximal ends as recipient vessels, and one patient having anterior serratus branch of the thoracodorsal vessels at the distal and proximal ends. All drainage tubes were removed within 7 to 10 days after surgery. Patients were discharged. Follow-up period ranged from 1 to 15 months, averaging 6 months. The patients recovered well postoperatively, with no flap-related complications occurring. All four patients were satisfied with the result of the reconstruction.Conclusion:The simultaneous application of the proximal and distal ends of the thoracodorsal artery and vein can ensure the safety of flap survival while reducing damage to the ribs and intercostal muscles, achieving better aesthetic result.

Objective:Explore the feasibility and advantages of using proximal and distal ends of thoracodosal artery and vein as recipient vessels in deep inferior epigastric perforator flap immediate breast reconstruction.Methods:The clinical data of patients who underwent breast reconstruction surgery using the proximal and distal ends of the thoracodorsal vein as recipient vessels at the Department of Breast and Thyroid Surgery of the Second Affiliated Hospital of Hainan Medical University from March 2022 to June 2023 were analyzed retrospectively. Preoperative examinations included thoracoabdominal angiography and color Doppler ultrasonic localization of the main trunk and perforators of the inferior epigastric vessels. The procedure began with mastectomy and axillary lymph node dissection, followed by the isolation of bilateral perforators and the main trunk of the abdominal flap. The main trunks of the bilateral inferior epigastric arteries were then transected, and their vascular pedicles exposed and anastomosed respectively to the proximal and distal ends of the thoracodorsal artery and vein. Both arteries and veins were joined end-to-end. The flap after trimming and reconstruction was then implanted into the cavity left after mastectomy through the incision. Breast positioning was performed with the patient in a knee-bent and hip-flexed position. After adjusting the shape of the reconstructed breast. The donor site was closed, the umbilicus was reconstructed, drainage tubes were placed, and the breast incision was closed. Postoperative follow-up monitored complications associated with the flap and patient satisfaction with the breast reconstruction, utilizing a self-assessment method.Results:Four female patients were included, aged (46.0±6.5) years, ranging from 37 to 52 years. All four patients had bilateral vascular pedicles in the donor area, with three patients having thoracodorsal vessels at the distal and proximal ends as recipient vessels, and one patient having anterior serratus branch of the thoracodorsal vessels at the distal and proximal ends. All drainage tubes were removed within 7 to 10 days after surgery. Patients were discharged. Follow-up period ranged from 1 to 15 months, averaging 6 months. The patients recovered well postoperatively, with no flap-related complications occurring. All four patients were satisfied with the result of the reconstruction.Conclusion:The simultaneous application of the proximal and distal ends of the thoracodorsal artery and vein can ensure the safety of flap survival while reducing damage to the ribs and intercostal muscles, achieving better aesthetic result.

OBJECTIVE: To investigate the clinical and prognostic characteristics of primary acute myeloid leukemia (AML) with 11q23/KMT2A rearrangements. METHODS: Clinical data of 90 patients with primary AML and 11q23/KMT2A rearrangements were analyzed retrospectively. RESULTS: By karyotyping analysis, 80 of the 90 patients had translocations involving 11q23/KMT2A, with t(9;11)(p22;q23), t(6;11)(q27;q23), t(10;11)(p12;q23) and t(11;19)(q23;p13) being the most common ones, while 10 cases were found to have non-translocation abnormalities. The overall complete remission (CR) rate was 75.6%, and patients with t(6;11) had lower CR rate compared with non-t(6;11) patients (47.1% vs. 82.2%, P = 0.005). After a median follow-up of 24.5 months, the patients receiving allo-hematopoietic stem cell transplantation (allo-HSCT) had significantly higher 3-year overall survival (OS) (80.3% vs. 16.6%, P < 0.001) and 3-year event-free survival (EFS) (73.5% vs. 16.3%, P < 0.001) compared with non-transplant patients. Patients with t(6;11) had the lowest 3-year OS (11.8% vs. 56.0%, P < 0.001) and 3-year EFS (5.9% vs. 53.8%, P < 0.001) compared with other type of abnormalities. No significant difference was noted in the survival between patients with t(9;11) and non-t(9;11) regardless whether they had received HSCT. CONCLUSION The clinical characteristics of primary AML with 11q23/KMT2A rearrangements are heterogeneous. Patients did not receive HSCT had poorer survival, particularly with the presence of t(6;11). Allo-HSCT could significantly improve the survival of such patients.
Humans ; Retrospective Studies ; Leukemia, Myeloid, Acute/therapy* ; Translocation, Genetic ; Gene Rearrangement ; Prognosis

Objective:To study the effects of modified citrus pectin (MCP) on the viability and gene expressions of synovial fibroblasts (SF) as well as SF treated by galectin-3 (Gal-3).Methods:Rabbit SF was isolated and cultured in vitro. Then SF was treated with different concentrations of MCP (0, 250, 500, and 750 mg/L). In addition, SF was further treated with the same different concentrations of MCP after treatment with 10 μg/ml Gal-3 for 24 h. The viability of SF was detected by CCK-8 on the first, third, and fifth day after treatment. The mRNA expression of transforming growth factor-β1 (TGF-β1), type I collagen (COL1A2), and Gal-3 in SF was detected by real-time quantitative PCR. The synthesis of type I collagen in SF was investigated by immunofluorescence staining. Results:MCP, especially at a concentration of 500 mg/L can inhibit the proliferation of SF significantly (all P < 0.05) on the first, third, and fifth day after treatment. Compared with the control group, MCP at different concentrations induced different gene expression profiles. In particular, MCP at high concentrations can upregulate the expression of TGF-β1, COL1A2 and Gal-3 in SF. However, MCP shows no significant effect on the synthesis of type I collagen in SF. MCP can down-regulate the expression of TGF-β1, COL1A2, and significantly reduce the synthesis of type I collagen in SF after Gal-3 treatment. Particularly, the effect of MCP at a concentration of 500 mg/L on inhibiting the expression of TGF-β1, COL1A2, and Gal-3 in SF is significant. Conclusions:MCP can inhibit the excessive proliferation of SF and regulate gene expression in SF.

Objective:To investigate the expression of SET-NUP214 fusion gene in hematological malignancies and to analyze its related clinical biological characteristics.Methods:The clinical data of 24 patients with SET-NUP214 fusion gene-positive hematological malignancies were retrospectively analyzed, and the Kaplan-Meier method was used for survival analysis.Results:Among the 24 patients with SET-NUP214 fusion gene, 15 cases of acute lymphoblastic leukemia (ALL) (13 cases of T-ALL and 2 cases of B-ALL) , 7 cases of acute myeloid leukemia (AML) , and 2 cases of T/myeloid mixed acute leukemia have been identified. The immunophenotype of 13 cases of T-ALL was mainly characterized by CD3 +CD2 -, 73.3% of ALL was characterized by myeloid marker expression, and 85.7% of AML was characterized by CD7 expression. Complete remission (CR) was achieved in 22 patients (91.7%) after induction chemotherapy. All 24 patients received allogeneic hematopoietic stem cell transplantation (HSCT) . With a median follow-up of 24 months, the 3-year relapse free survival (RFS) of AML and ALL was 85.7% and 33.3%, respectively ( P=0.128) . Comparing 13 cases of SET-NUP214-positive and 62 cases of SET-NUP214-negative T-ALL, the CR rates of induction chemotherapy were 92.3% and 93.5% ( P=0.445) , and the 4-week CR rates of induction chemotherapy were 69.2% and 72.6%, respectively ( P=0.187) ; the differences were not statistically significant. After HSCT, the 3-year RFS of SET-NUP214 +T-ALL and SET-NUP214 -T-ALL was 38.5% and 66.4%, respectively ( P=0.028) , and the difference was statistically significant. Conclusion:The SET-NUP214 fusion gene is mainly detected in T cell-derived hematological malignancies, and the prognosis of SET-NUP214 positive T-ALL is relatively poor.

OBJECTIVE: To trace a rare case of chronic myeloid leukemia (CML) with a four-way Philadelphia chromosome variant by cytogenetic analysis in order to provide a basis for the selection of treatment. METHODS: Bone marrow morphology, chromosomal karyotyping, fluorescence in situ hybridization (FISH) and real-time quantitative PCR (RQ-PCR) were used for the diagnosis and staging of the disease. Point mutations in the tyrosine kinase domain of ABL1 gene were detected by Sanger sequencing. RESULTS: The patient was initially diagnosed as CML in chronic phase (CML-CP) with a chromosomal karyotype of 46,XX,t(5;9;22;6)(q13;q34;q11;q25), while FISH revealed presence of a variant Philadelphia chromosome translocation. Clonal evolution has occurred after 38 months of tyrosine kinase inhibitor (TKI) treatment, when cytogenetic analysis revealed coexisting t(5;9;22;6)(q13;q34;q11;q25) and t(5;9;22;6;17)(q13;q34;q11;q25;q11). After 57 months of TKIs treatment, only the t(5;9;22;6;17) clone was detected. Three months later, hyperdiploidy with additional abnormalities were detected in addition to t(5;9;22;6;17). Three mutations, including p.Tyr253Phe, p.Thr315Ile and p.Gly250Glu, were identified in the tyrosine kinase domain of the ABL1 gene during the course of disease. The patient did not attain cytogenetic and molecular response to TKIs. CONCLUSION The four-way variant translocation may be genetically unstable. Clonal evolution and genetic mutations are likely to occur during TKIs treatment, resulting in poor response to drug therapy. This observation, however, needs to be confirmed by large-scale studies.
Enzyme Inhibitors/therapeutic use* ; Evolution, Molecular ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive/genetics* ; Mutation/genetics* ; Philadelphia Chromosome ; Translocation, Genetic

Objective: To analyze the characteristics of myeloid neoplasms with t (3;21) (q26;q22) . Methods: Clinical data of patients with t (3; 21) (q26; q22) , diagnosed as hematologic malignancies in Peking University people's hospital from January 2011 to March 2018, were collected retrospectively. 19 patients in our hospital and forty-eight patients bearing t (3;21) (q26;q22) with detailed survival data reported in literature were summarized. Kaplan- Meier method was used for survival analysis. Results: Among 19 patients, including 15 males and 4 females with a median age of 36 years (22-68 years) , 4 cases was diagnosed as de novo acute myeloid leukemia (AML) , 4 as myelodysplastic syndromes (MDS) , 3 as MDS-AML and 8 as chronic myelogenous leukemia (CML) in myeloid blast transformation. All of the 19 patients were detected to have t (3;21) (q26;q22) by G-banding technique and 13 carried additional cytogenetic aberrations. 9 of the 19 patients were detected for positive AML1-MDS1 fusion genes. In the 9 patients with detailed follow-up data, 6 patients received chemotherapy and only 2 achieved complete remission (CR) while 4 with no response. During the follow-up period, 8 patients died and the median overall survival (OS) was 6 months (4.5 to 22 months) . Survival analysis of the present 9 patients together with the literature data showed that the prognosis was poor and the median OS was 7 months. In particular, AML/t-AML had the worst prognosis. Hematopoietic stem cell transplantation (HSCT) could significantly improve survival, the median OS in HSCT group and non-HSCT group were 20.9 and 4.7 months respectively (P<0.001) . Conclusions t (3; 21) (q26; q22) is a rare recurrent chromosomal abnormality which is detected mainly in myeloid neoplasm and confer to poor clinical prognosis. HSCT should be recommended to improve the outcomes.

OBJECTIVETo report on a case of therapy-related acute monocytic leukemia(t-AML) with t(11;17) (q23;q21)/MLL-AF17q after successful treatment for acute promyelocytic leukemia(APL) with t(15;17) (q22;q21)/PML-RARα.

METHODSA MICM method (bone marrow morphology(M), immunophenotype(I), cytogenetics(C), and molecular biology(M)) was used for the diagnosis and classification of the disease at the time of onset and transformation.

RESULTSThe patient was initially identified with typical morphology and immunophenotype of APL. She has carried t(15;17)(q22;q21) and PML-RARα fusion gene but was without t(11;17)(q23;q21) or MLL gene abnormalities. After 13 months of successful treatment, she has transformed to AML with typical morphology and immunophenotype. t(11;17)(q23;q21) and MLL-AF17q fusion gene were detected in her bone marrow sample, while no PLZF-RARα fusion gene was detected by real-time quantitative reverse-transcription PCR(RQ-PCR) and fluorescence in situ hybridization(FISH).

CONCLUSIONt-AML is a serious complication after successful treatment of APL. t(11;17)(q23;q21) is not specific for the diagnosis of variant APL and can also be detected in t-AML. RQ-PCR and FISH are essential for the diagnosis of such patients.

Chromosomes, Human, Pair 11 ; Chromosomes, Human, Pair 15 ; Chromosomes, Human, Pair 17 ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Leukemia, Monocytic, Acute ; genetics ; Leukemia, Promyelocytic, Acute ; genetics ; Middle Aged ; Neoplasms, Second Primary ; genetics

Objective: To compare the cytogenetic response detected by conventional banding analysis (CBA) and fluorescence in situ hybridization (FISH) and to explore the correlation between the cytogenetic and molecular response in chronic myeloid leukemia (CML) patients during tyrosine kinase inhibitor (TKI) treatment. Methods: CBA, FISH and real-time quantitative reverse transcriptase polymerase chain reaction (RQ-PCR) methods were performed to detect the cytogenetic and molecular response simultaneously in 504 bone marrow samples from 367 CML patients who received TKI treatment. Results: Among 504 samples, 344 were detected to reach complete cytogenetic response (CCyR) by CBA, while 297 samples reached CCyR by FISH which were considered to carry BCR-ABL positive cells<1%. When the results of CBA, FISH and RQ-PCR were compared in 493 samples at the same time, it showed that in 337 samples with CBA-CCyR, 273 (81.0%) reached FISH-CCyR and 289 (85.8%) were BCR-ABL^IS (International Scale, IS) ≤1% by RQ-PCR, compared to 9.0 (261/290) were BCR-ABL^IS ≤1% among 290 samples with FISH-CCyR. There was no significant difference in the median value of BCR-ABL^IS between samples in CBA-CCyR and FISH-CCyR (0.21% vs 0.13%, z=-1.875, P=0.061) . Furthermore, when the samples were divided into three groups according to BCR-ABL positive cells (0,>0~<1%, 1%~5%) by FISH, the statistical difference was observed, the proportion of samples with BCR-ABL^IS ≤1% in the three groups were 94.1%, 57.6% and 27.7% respectively (χ²=43.499, P<0.001; χ²=9.734, P=0.003) , while the median value of BCR-ABL^IS were 0.10%, 0.64% and 1.80% respectively (z=-5.864, P<0.001; z=-4.787, P<0.001) . Conclusion FISH results were in good concordance with CBA in identify samples in CCyR, FISH was more sensitive and had better correlation with RQ-PCR results than CBA, but how to define FISH-CCyR need further study.