[Expression of Th17 and IL-23 in Peripheral Blood and Their Relationship with Immunophenotype in Patients with Acute Myeloid Leukemia].

OBJECTIVE: To observe the expression of helper T cells 17(Th17), interleukin 23 (IL-23) in peripheral blood in patients with acute myeloid leukemia (AML), to analyze the relationship between Th17, IL-23 in peripheral blood and immunophenotype. METHODS: 105 patients with AML in the hospital from January 2019 to January 2021 were prospectively selected as the research subjects, the expression of Th17 and IL-23 in peripheral blood of patients with AML was detected by flow cytometry; immunophenotype was detected and counted. The relationship between the expression of Th17, IL-23 in peripheral blood and immunophenotype of AML patients was analyzed. Draw ROC curve and analyze the predictive value of Th17 and IL-23 expression in peripheral blood to immunophenotype. RESULTS: The immunophenotype results of AML patients showed that myeloid antigen, lymphoid antigen and hematopoietic stem/progenitor cell marker antigen were positive expressed for various antigens in 105 AML patients, in myeloid antigens, CD13+ accounted for the highest proportion (93.33%), in lymphoid antigens, CD56+ accounted for the highest proportion (32.38%), and in hematopoietic stem/progenitor cell marker antigens, CD38+ accounted for the highest proportion (68.57%). The expression of Th17 in peripheral blood of AML patients with CD56+, CD7+, CD34+ and human leukocyte antigen DR+(HLA-DR+) were higher than that of AML patients with CD56-, CD7-, CD34-, HLA-DR-, the expression of IL-23 in peripheral blood of AML patients with CD56+, CD34+ and HLA-DR+ were higher than that of AML patients with CD56-, CD34-, HLA-DR-, the differences were statistically significant (P<0.05); compared the expression of Th17 and IL-23 in peripheral blood between other antibody positive and negative patients, there was no statistical significant difference (P>0.05). Logistic regression analysis showed that the high expression of Th17 in patients with AML was related to the positive expression of CD56, CD7, CD34 and HLA-DR in the detection of immunophenotype, the high expression of IL-23 was related to the positive expression of CD56, CD34 and HLA-DR in the detection of immunophenotype. The ROC curve showed that the AUC of expression levels of Th17 and IL-23 in peripheral blood alone and in combination for predicting CD56+, CD34+, HLA-DR+ and Th17 in peripheral blood for predicting CD7+ were mostly 0.5-0.7, which had certain predictive value, but the predictive performance was low. CONCLUSION: Myeloid antigen, lymphoid antigen and hematopoietic hematopoietic stem/progenitor cell marker antigen are positive expressed for various antigens in AML patients, the high expression of Th17 in peripheral blood of AML patients is related to the positive expression of CD56, CD7, CD34 and HLA-DR in detection of immunophenotyping, the high expression of IL-23 is related to the positive expression of CD56, CD34 and HLA-DR in the detection of immunophenotype.

Management of chronic myeloid leukemia presenting with isolated thrombocytosis and complex Philadelphia chromosome: A case report.

RATIONALE: Chronic myelogenous leukemia (CML) with thrombocytosis and complex chromosomal translocation is extremely rare in clinical setting. Here, we reported the clinical and pathological characteristics of CML patients, which were characterized by thrombocytosis and complex Philadelphia chromosome translocation. Moreover, we also introduced our therapeutic schedule for this patient as well as review relative literature. PATIENT CONCERNS: A 24-year-old female presented with night sweating, fatigue, and intermittent fever for 1 month. DIAGNOSIS: Fluorescence in situ hybridization results revealed that breakpoint cluster region (BCR)-Abelson (ABL) gene fusion in 62% of the cells and karyotyping showed a complex 3-way 46, XY, t(9;22;11) (q34;q11;q13) [19/20] translocation. This patient was diagnosed with CML complicated with thrombocytosis and complex Philadelphia chromosome translocation. INTERVENTIONS: The patients received continuously oral imatinib mesylate tablets (400 mg) once a day. OUTCOMES: After treatment with imatinib for 3 months, the BCR/ABLIS was less than 0.1% and achieved major molecular response. Moreover, the BCR/ABLIS of this patient achieved major molecular response. The BCR/ABLIS values at 6 months and 12 months were less than 0.01% and 0.0032%, respectively. And no BCR/ABL fusion was detected in the next 2 years follow-up period. LESSONS: Imatinib might represent a preferred therapeutic option for CML patients with rare thrombocytosis and complex chromosomal translocation. In addition, BCR/ABL fusion gene examination in patients with thrombocytosis might represent an effective strategy to avoid the misdiagnosis of this specific CML population.

[Expression Level and Clinical Value of PTEN Gene in Patients with Acute T Lymphocytic Leukemia].

OBJECTIVE: To study the PTEN gene expression level and its clinical significance in patients with acute T lymphoblastic leukemia. METHODS: One hundred and twenty-four patients with T-ALL treated in our hospital from January 2014 to May 2018 were selected, and 120 healthy people were selected as control group. The bone marrow of the patients were collected, and mononuclear cells were separated out. PTEN gene expression level was detected by RT-PCR, PTEN protein expression level was detected by Western blot, Kaplan-Meier was used to analyze the survival rate of patients with T-ALL, Cox multivariate regression analyzed was used to analyze the independent influencing factors of patients, and the correlation between PTEN level and clinical characteristics of patients with T-ALL and its prognostic value were analyzed. RESULTS: The relative level of PTEN gene in patients with T-ALL was 0.19±0.06, which was significantly lower than that in control groups (P<0.05). There was significantly positive correlation of the expression level of PTEN gene with white blood cell count （r=0.993）and peripheral blood immature cells （r=0.996） in patients with T-ALL. There was no correlation between PTEN gene expression level and sex, age, LDH level, Hb level, platelet count in patients with T-ALL. And it was found that expression levels PTEN protein in the middle-risk group, the standard-risk group and the high-risk group were significant lower than those in the control group (P<0.05), while those in the high-risk group were significantly lower than those in other groups (P<0.05). Kaplan-Meier survival analysis showed that OS and DFS in PTEN gene high expression group were higher than those in PTEN low expression group (P<0.05). Cox multivariate regression analysis showed that white blood cell count and PTEN gene were independent influencing factors of OS (P<0.05); platelet count and PTEN gene were independent influencing factors of DFS (P<0.05). CONCLUSION: PTEN gene level relates to the prognosis of patients with T-ALL. Patients with better prognosis show a higher PTEN gene level, which provides reference for clinical treatment of patients with T-ALL.

Subculturing cells have no effect on CRISPR/Cas9-mediated cleavage of UL30 gene in pseudorabies virus.

CRISPR/Cas9-mediated genome editing can inhibit virus infection by targeting the conserved regions of the viral genomic DNA. Unexpectedly, we found previously that pseudorabies virus (PRV) could escape from CRISPR/Cas9-mediated inhibition. In order to elucidate whether the escape of PRV from Cas9-mediated inhibition was due to cell deficiencies, such as genetic instability of sgRNA or Cas9 protein, the positive cells were passaged ten times, and PRV infection in the sgRNA-expressing cells was evaluated in the present study. The results showed that subculturing cells has no effect on Cas9-mediated cleavage of PRV. Different passages of PX459-PRV cells can stably express sgRNA to facilitate Cas9/sgRNA cleavage on the UL30 gene of PRV, resulting in a pronounced inhibition of PRV infection. Studies to elucidate the mechanism of PRV escape are currently in progress.

Clinical value of multi-slice spiral computed tomography angiography and three-dimensional reconstruction in the diagnosis of double aortic arch.

The aim of this study was to evaluate the clincal value of multi-slice spiral computed tomography (MSCT) in the diagnosis of double aortic arch (DAA) and analyze the associated angiography and 3D reconstruction imaging to increase the accuracy of DAA diagnosis. Clinical and imaging data from 15 patients diagnosed with DAA by MSCT were summarized and compared with the corresponding surgical observations. The DAA diagnosis by MSCT for the 15 patients was confirmed by the surgical observations. A total of 13 cases were categorized as type I (double arches are open), including nine with a larger right arch, two with a larger left arch and two with balanced arches. Two cases were categorized as type II (one atretic arch), characterized by left atretic arch. Among the 15 patients, ultrasound diagnosis succeeded in nine cases and failed in the rest. A single malformation was identified in only two cases, whereas the rest had single or multiple combination(s) of intracardiac and extracardiac malformations, including seven with different levels of tracheobronchial stenosis. MSCT was demonstrated to precisely diagnose DAA complicated by malformation and tracheobronchial stenosis. MSCT is an essential therapeutic strategy and serves as a primary method in DAA diagnosis.

Diagnosis of congenital aortic arch anomalies in chinese children by multi-detector computed tomography angiography.

The purpose of this study was to evaluate the value of multi-detector computed tomography (MDCT) angiography for the diagnosis of congenital aortic arch anomalies and present the radiological images of congenital aortic arch anomalies in Chinese children. MDCT angiography and transthoracic echocardiography (TTE) were applied for the diagnosis of congenital aortic arch anomalies in 362 Chinese children between May 2006 and December 2011 (age ranges from 5 days to 12 years; mean age, 3.3 years). Surgery and/or catheter angiography (CA) were conducted in all patients to confirm the final diagnosis. In the 362 Chinese children with congenital heart anomalies, congenital aortic arch anomalies were definitely diagnosed in 198 children and 164 children ruled out by operation and/or (CA). Among the 198 children with anomalies, coarctation of aorta (CoA), interruption of aortic arch (IAA), right aortic arch, aberrant right subclavian artery and double aortic arch were diagnosed in 134, 32, 20, 10 and 2 children respectively, and there were 6 cases with uncommon congenital aortic arch anomalies: 2 had double aortic arch including 1 with five branches of the aortic arch, 2 had isolation of the right subclavian artery with two patent ductus arteriosus (PDA), 1 had an isolation of the common carotid artery with a PDA, and 1 had double PDA with a single ventricle and pulmonary artery atresia. Among the 32 children with IAA, 28 were of type A, and 4 were of type B. The diagnostic sensitivity, specificity and accuracy of MDCT angiography for congenital aortic arch anomalies were 100% (198/198), 98% (161/164) and 99% (359/362), respectively. The diagnostic sensitivity, specificity and accuracy of TTE were 92% (182/198), 81% (133/164) and 87% (315/362), respectively. In conclusion, MDCT angiography is a reliable, noninvasive imaging technique for the diagnosis of congenital aortic arch anomalies in children. Sometimes, even more information can be obtained from this technique than from conventional angiography.

[Pneumocephalus caused by neonatal Enterobacter cloacae infection in a case].

OBJECTIVE: Gas-containing encephalo-meningitis is very rare. There have only been a few cases reported in the literature. The majority of neonatal cases reported in literature died. We report a case of a 5-day-old neonate who developed diffuse pneumocephalus from Enterobacter cloacae septicemia and intracranial infection. METHOD: This neonate was initially treated with penicillin and mezlocillin. He also received phototherapy, intravenous infusion, correction of acidosis and other supportive therapy. Complete blood count, C-Reactive protein, cranial CT scan, blood culture, cerebrospinal fluid culture and biochemistry were tested repeatedly. RESULTS: This neonate's condition deteriorated after admission. He developed respiratory distress, increased muscle tone and decreased level of consciousness. His WBC and C-reactive protein were elevated, while blood gas, electrolytes, liver enzymes and renal function were within normal range initially. Cranial CT scan was done which demonstrated diffuse pneumocephalus. He was transferred to a higher level hospital for further management at the request of the family. Blood culture done in our hospital subsequently showed growth of Enterobacter cloacae. The infant developed seizures and further deterioration in level of consciousness after transfer. Antibiotics were switched to penicillin and ceftizoxime. Cranial CT scan repeated 2 days after transfer showed hydrocephalus and some resolution of pneumocephalus. Cerebrospinal fluid (CSF) was aspirated from the lateral ventricles two weeks after admission. CSF culture also showed growth of Enterobacter cloacae. Antibiotic was switched to imipenem according to antibiotic sensitivity. His general condition was improved. Blood and CSF cultures were negative 1 month after admission. His head circumference at discharge was 34.6 cm. Repeat cranial CT scan at 4 month of age demonstrated severe hydrocephalus, diffuse leukomalacia and calcification. This infant suffered significant neurodevelopmental deficit. Muscle tone was diffusely increased. Head circumference at 9 month of age was 48.4 cm. CONCLUSION: This case suggests the importance of Enterobacter cloacae infection in the newborns. Our analysis of 34 cases of Enterobacter cloacae infection showed that 93.75% - 100% were sensitive to quinolones, 94.12% were sensitive to imipenem, 73.33% were sensitive to gentamicin, 50% were sensitive to piperacillin-tazobactam. Enterobacter cloacae is generally not sensitive to penicillin, first and second generation cephalosporins (0 - 21.4%). Enterobacter cloacae septicemia and intracranial infection in neonates have a high mortality rate and can result in severe neurodevelopmental deficit in survivors.