OBJECTIVE: To develop a multiplex allele-specific PCR (AS-PCR) assay with three-color fluorescence labeling for mitochondrial DNA (mtDNA) SNP typing. METHODS: Based on the principle of AS-PCR, the primer sets were designed for 20 SNP located on the coding region of mtDNA and divided into 2 groups labeled with FAM and HEX fluorescence, respectively. A primer set included two forward (reverse) allelic specific primers with different sizes and a generic reverse (forward) primer. Blood samples from 200 unrelated individuals were analyzed by AS-PCR and capillary electrophoresis. Three random samples at least for each SNP site were examined and verified by direct sequencing. The haplotype frequency was investigated. RESULTS: Distinct electropherograms of 200 blood samples were obtained successfully. The typing results of direct sequencing were identical to those obtained from AS-PCR. The minimum detectable DNA concentration was 0.2 pg under the system of 10 microL. The sensitivity of the DNA concentrations ranged from 0.5 to 5 pg. The 200 individuals were assigned into 15 haplotype, and the haplotype diversity was 0.906 0. CONCLUSION AS-PCR is a simple, rapid and efficient method for mtDNA SNP typing, and can be applied to forensic practice.
Alleles ; DNA ; DNA Primers ; DNA, Mitochondrial/analysis* ; Electrophoresis, Capillary ; Haplotypes ; Humans ; Mitochondria ; Polymerase Chain Reaction/methods* ; Polymorphism, Single Nucleotide ; Sequence Analysis, DNA

OBJECTIVETo detect the expression of the fusion genes resulting from chromosome abnormalities in childhood acute lymphoblastic leukemia(ALL) and its conformity to WHO classification.

METHODSSixty-two children with ALL were investigated. The expression of fusion genes was determined by multiplex reverse transcription-polymerase chain reaction (RT-PCR), karyotyping (R band) and immunophenotyping (by flow cytometry) were also performed.

RESULTSOf the 62 patients, 23(37.1%) were found to carry 13 different fusion genes. The patients with immunophenotype of Pre-B-ALL were found to carry: TEL/AML1(3 cases); E2A/PBX1, E2A/HLF, TLS/ERG, MLL/AF4, MLL/AF9, MLL/AF10, MLL/AFX-MLL/AF6-MLL/ELL, MLL/AF6-MLL/ELL, dupMLL (one case for each); and HOX11 (6 cases). The patients with immunophenotype of Pre-T-ALL were found to carry: TAL1D (4 cases, one is also found to have HOX11 expression); and HOX11 (2 cases). The multiplex RT-PCR in combination with chromosome analysis revealed genetic abnormalities in 69.4%(43/62) of childhood ALL.

CONCLUSIONMultiplex RT-PCR combined with chromosome analysis and immunophenotyping can provide reliable and helpful information for the diagnosis, therapy evaluation and prognosis prediction in childhood ALL, which may also serve as a basis on which to implement the criteria of WHO classification.

Adolescent ; Child ; Child, Preschool ; Chromosome Aberrations ; Core Binding Factor Alpha 2 Subunit ; genetics ; metabolism ; DNA-Binding Proteins ; genetics ; metabolism ; Flow Cytometry ; Homeodomain Proteins ; genetics ; metabolism ; Humans ; Immunophenotyping ; Infant ; Karyotyping ; Myeloid-Lymphoid Leukemia Protein ; genetics ; metabolism ; Oncogene Proteins, Fusion ; genetics ; metabolism ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; metabolism ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA-Binding Protein FUS ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factors ; genetics ; metabolism

OBJECTIVETo study the value of combination assay of multiplex RT-PCR and karyotypic analysis in the diagnosis and classification of childhood acute lymphoblastic leukemia (ALL).

METHODSFifty cases of childhood ALL patients were studied by multiplex RT-PCR in combination with R or G banding karyotype analysis.

RESULTSOf the 50 childhood ALL patients, 18 (36.0%) carried 11 types of fusion genes including E2A/PBX1, TEL/AML1, TLS/ERG, MLL/AF4, MLL/AF9, MLL/AF10, MLL/AFX, MLL/AF6, MLL/ELL, TAL1D, and HOX11, revealed by multiplex RT-PCR, and in 48 cases, 24 (57.1%) had chromosome abnormalities. Among the latter, numeral chromosome abnormalities and chromosome deletions accounted for 75.0% (18/24), while translocations 25.0% (6/24). The multiplex RT-PCR in combination with chromosome analysis could detect genetic abnormalities in 70% (35/50) of childhood ALL.

CONCLUSIONSMultiplex RT-PCR combined with chromosome analysis can enhance the detection rate of genetic abnormalities in childhood ALL. It provides reliable evidence for the diagnosis, classification and prognosis.

Adolescent ; Child ; Child, Preschool ; Chromosome Aberrations ; Female ; Humans ; Infant ; Karyotyping ; Male ; Oncogene Proteins, Fusion ; genetics ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; classification ; diagnosis ; genetics ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

OBJECTIVETo investigate the interrelations among morphology, immunology, cytogenetics and clinical outcome in childhood acute leukemia with 11q23 abnormalities.

METHODSEighteen patients with 11q23 abnormalities, from 320 childhood acute leukemia patients, were retrospectively analysed for cell morphology, flow cytometry, immunophenotyping, R-banding karyotype as well as clinical features and prognosis. Twenty cases of childhood AL with normal karyotype during the same period were used as control.

RESULTSThe incidence of 11q23 abnormalities in our childhood acute leukemia patients was 5.63% including 14 acute lymphoblastic leukemia (ALL) and 4 acute myeloid leukemia (AML). Of 16 cases immunophenotypically tested, 13 expressed lymphoid antigens and 3 CD(34) and other myeloid antigens. Karyotype analysis disclosed the following abnormalities: t(4; 11)(q21; q23) in 6 cases, t(10; 11)(p13; q23) in 3, t(11; 19)(q23; p13) in one and del(11)(q23) in 6. The complete remission rate for these patients with 11q23 abnormalities was comparable to that of the control (72.2% vs 80.0%, P > 0.05), while the mortality rate in the former was significantly higher than that in the latter (61.1% vs 25.0%, P < 0.05).

CONCLUSIONS11q23 abnormalities were mainly seen in childhood ALL and acute monocytic leukemia with unique prognostic features.

Acute Disease ; Adolescent ; Child ; Child, Preschool ; Chromosome Aberrations ; Chromosomes, Human, Pair 11 ; genetics ; Cytogenetic Analysis ; Female ; Humans ; Immunophenotyping ; Infant ; Leukemia ; drug therapy ; genetics ; immunology ; Male ; Prognosis ; Retrospective Studies

OBJECTIVEWith the improvement of the diagnosis and treatment, the complete remission (CR) rate and the survival rate of childhood acute lymphoblastic leukemia have been increased in the recent 10 years. The objective of this study was to analyze the outcomes of 119 standard-risk childhood acute lymphoblastic leukemia (SR-ALL) patients, and explore how to improve the survival rate in ALL.

METHODSA total of 119 patients aged 14 months to 15 years were diagnosed as SR-ALL according to the Suggestion of Diagnosis And Treatment for Childhood Acute Leukemia-1993. Among them, seventy-nine were boys and 40 were girls. All of the patients were treated with the CCLG-97 protocol and were followed up for a period of 20 approximately 78 months.

RESULTSThe complete remission rate reached 97.4% in four-week induction. Twenty-one patients were out of follow-up, comprising 63%, 14%, 10%, 8% and 5% of all subjects in 1998, 1999, 2000, 2001 and 2002, respectively. The overall survival rates were 93.3%, 90.2%, 88.0%, 85.0%, 85.0% and 85.0% in 1 year, 2 years, 3 years, 4 years and 5 years, respectively. Relapses occurred in 13 patients (13.8%). Among 9 isolated hematologic relapses, 5 patients (56%) were given irregular therapy, 2 did not reach CR within 4 weeks and relapsed 2 years later, 2 accepted regular therapy, 1 was of hypodiploidy and 1 T-ALL. Isolated central nervous system (CNS) relapse occurred in 4 patients (4.3%). Fifteen patients (12.6%) died, 5 of whom (4.2%) died of complications.

CONCLUSIONReinforcing administration and regular therapy are important to improve the long-term survival rate in childhood ALL. The clinical classification should be adjusted with the improvement of diagnostic methods. CCLG-97 protocol decreased the rate of the relapses in SR-ALL and didn't increase the rate of therapy-related death. High-dose methotrexate should be used in therapy and its dosage, usage and individualized therapeutic regimen should be further studied.

Adolescent ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Child ; Child, Preschool ; China ; Disease-Free Survival ; Female ; Follow-Up Studies ; Humans ; Infant ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; mortality ; prevention & control ; Remission Induction ; methods ; Risk Factors ; Secondary Prevention ; Survival Rate ; Time Factors ; Treatment Outcome

OBJECTIVE With Poloxamer 407(P407) as the carrier, doxorubicin-copper-curcumin ternary complex was encapsulated in the micelles to achieve quantitative and intelligent drug release behavior and synergistic effect.METHODS With P407 as the carrier,the preparing technology was optimized by Box-Behnken design and the in vitro pH-sensitive drug release behavior of DOX-Cu-Cur/P407 was examined;the MTT analysis was carried out to evaluate the cytotoxicity to tumor cells.RESULTS Under the optimum condition, the encapsulation efficiency of doxorubicin and curcumin was (93.17 ± 1.05)% and (100.03 ± 1.10)%, size distribution was (31.06 ± 4.20) nm,PDI is 0.174 ± 0.028.Meantime, in vitro drug release test showed the DOX-Cu-Cur/P407 micelles was pH-sensitive.The MTT analysis showed DOX-Cu-Cur/P407 micelles could significantly inhibit the tumor cells with a better synergistic effect compared with free drugs.CONCLUSION The optimal DOX-Cu-Cur/P407 micelle is uniform in size and has satisfactory encapsulation efficiency.Meanwhile it is sensitive to acidic microenvironment of tumor and can achieve the purpose of quantitative and intelligent drug release.

Objective:To investigate the effects of Huber 360 neuromuscular control training and evaluation system (Huber 360) on balance dysfunction for patients with stroke. Methods:From January to November, 2020, 16 patients with stroke were randomly assigned to control group (n = 8) and observation group (n = 8). Both groups received routine motor function training, while the observation group accepted Huber 360 training in addition, for three weeks. They were assessed with Berg Balance Scale, Huber 360, 10-meter Walking Test and Timed 'Up & Go' Test before and after training. Results:All the indexes improved after training in both groups (t > 1.368, P < 0.05), and improved more in the observation group than in the control group (t > 2.230, P < 0.05). Conclusion:Huber 360 can promote the recovery of balance for stroke patients.

Objective To explore the role and mechanism of stabilizing microtubules of endothelial cells and pericytes for ameliorating the dysfunction of microvasculature after spinal cord injury(SCI). Methods The endothelial cells and pericytes from rat brain microvascular tissue (microvessel) were separated and subjected to glucose oxygen deprivation (OGD). The cell viability was detected by CCK-8 and the expression of α-tubulin was detected by immunofluorescence and Western blotting. Rats (n = 36) were subjected to dorsal spinal cord transection at T

Objective To evaluate the pharmacokinetic characteristics and safety of single and multiple oral azvudine tablets in healthy young and elderly Chinese subjects.Methods This was a open-label and parallel-group study.The trial consisted of two groups:healthy young subjects group and healthy elderly subjects group,with 12 subjects in each group.Enrolled subjects were first given a single dose,fasting oral azvudine tablet 5 mg,after a 3-day cleansing period entered the multiple dose phase,fasting oral azvudine tablet 5 mg·d-1 for 7 days.Results After a single dose of azvudine 5 mg,Cmax and AUC0-∞ were(4.76±2.12)ng·mL-1,(6.53±2.20)ng·mL-1·h,and Tmax,t1/2 were 0.75,1.87 h in young subjects;Cmax and AUC0-∞ were(6.40±3.25)ng·mL-1,(9.50±3.70)ng·mL-1·h,and Tmax,t1/2 were 0.63,2.66 h in elderly subjects.After a multiple dose of azvudine 5 mg·d-1 for 7 d,Cmax and AUC0-∞ were(3.26±1.61)ng·mL-1,(5.38±2.19)ng·mL-1·h,and Tmax,ss,t1/2,ss were 0.88,2.13 h in young subjects;Cmax,ss and AUC0-∞,ss were(3.97±2.09)ng·mL-1,(6.71±3.26)ng·mL-1·h,and Tmax,ss,t1/2,ss were 0.75,2.56 h in elderly subjects.Elderly/young geometric mean ratios and 90％CIs were 128.37％(88.23％-186.76％),139.93％(105.42％-185.72％),140.03％(106.33％-184.41％)for azvudine Cmax,AUC0-t,AUC0-∞ after a single dose,and were 118.66％(80.83％-174.20％),118.41％(83.60％-167.69％),118.95％(84.78％-166.89％)for azvudine Cmax,AUC0-t,AUC0_∞ after a multiple dose of azvudine 5 mg·d-1 for 7 d.Conclusion After single and multiple oral administration of azvudine tablets,systemic exposure to azvudine was higher in healthy elderly subjects compared with healthy young subjects.After taking azvudine tablets,the types,severity and incidence of adverse events and adverse drug reactions in healthy elderly people were not significantly different from those in healthy young subjects.Azvudine was found to be safe and well tolerated in healthy elderly subjects.

Objective:To explore the preventive measures of nasal pressure injury in patients with preserved nasal endotracheal intubation after oral and maxillofacial neoplasms resection and simultaneous free tissue flap transplantation.Methods:Using the convenient sampling method, a total of 520 patients who underwent oral and maxillofacial neoplasms resection and simultaneous free tissue flap transplantation in Peking University Hospital of Stomatology were selected as the research objects from August 2018 to December 2019. According to the random number table method, they were divided into the experimental group and the control group. Patients in the control group received routine care, while in the experimental group, silk surgical tape was used to re-fix the endotracheal intubation tube away from the original position, and the special-shaped inflatable balloon was used to support the remote end of the tube. The incidence of nasal pressure injury, medical adhesive-related skin injury (MARSI) and the occurrence of complications caused by the fixation of tracheal intubation of patients were compared between the two groups. In the end, 246 patients in the experimental group and 248 patients in the control group completed the study.Results:No stage 3 or above nasal pressure injury occurred in the two groups. The incidence of nasal pressure injury related to nasotracheal tube in the experimental group was 2.85% (7/246) , lower than 12.10% (30/248) in the control group (χ 2=15.254, P<0.001) . There was no statistically significant difference in the incidence of MARSI of patients between the two groups ( P>0.05) . No complications caused by endotracheal intubation fixation occurred in both groups. Conclusions:The use of silk surgical tape to re-fix the endotracheal intubation and the combination of special inflatable balloon to support the end of the endotracheal intubation can prevent nasal pressure injury, and the safety is better.