OBJECTIVETo determine the feasibility of detecting target gene expression and evaluate the expression of homeobox gene HOX11L2 as well as its clinical significance in paraffin-embedded T-lymphoblastic lymphoma T-LBL.

METHODSHOX11L2 mRNA was analyzed by RT-PCR and real-time fluorescent quantitative PCR (RQ-PCR) on paraffin-embedded tissues in 54 cases of T-LBL.

RESULTSHOX11L2 expression was observed in 7 cases (13.0%). The age ranged from 5 to 15 years, with a median of 9 years. All of the HOX11L2 expressed patients had prior mediastinal masses, and 5 of them expressed high level of Ki67 antigen (PI> or =80%). Symptoms of CNS invasion were diagnosed in 2 cases. Significant differences in the age ratio (P < 0.05), the level of Ki67 antigen (P < 0.05) and the incidence of mediastinal involvement (P < 0.05) were observed between patients positive or negative for HOX11L2. Patients positive for HOX11L2 had a worse outcome than those negative for HOX11L2, with median survival of 7 and 11.5 month, respectively. No statistic difference was found in sex, clinical stages, CNS and bone marrow involvement between HOX11L2 positive and negative groups.

CONCLUSIONSDetection of target gene expression in paraffin-embedded tissues is feasible by RT-PCR or RQ-PCR. HOX11L2 expression was limited in childhood with T-LBL and probably associated with the development and poor prognosis of some pediatric T-LBL patients.

Feasibility Studies ; Homeodomain Proteins ; genetics ; metabolism ; Humans ; Paraffin Embedding ; Polymerase Chain Reaction ; methods ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; metabolism ; RNA, Messenger ; genetics

OBJECTIVETo study the clinicopathologic features, immunohistochemical findings and prognosis of precursor lymphoblastic lymphoma/acute lymphoblastic leukemia (LBL/ALL).

METHODSOne hundred and fifty-three cases of LBL/ALL were retrospectively analyzed. Immunohistochemical study was carried out. The pathologic findings were correlated with Ann Arbor tumor stage, Ki-67 index, other clinical parameters (including mediastinum/bone marrow involvement, hepato-splenomegaly, age and gender of the patients) and the survival data.

RESULTSStaining for TdT and CD99 was positive in 79.1% (121/153 cases) and 96.3% (131/136 cases), respectively. The cases were categorized into three groups according to the immunohistochemical findings, as follows: precursor T-cell, precursor B-cell and undefined. T-LBL/ALL accounted for 69.3% (106/153 cases) of all of the cases. The male-to-female ratio was 2.4:1 (including 75 males and 31 females). The median age at diagnosis was 17.5 years (ranged from 2 years to 68 years). Ninety-two patients (86.8%) presented with peripheral lymphadenopathy and 59 of them (55.7%) had mediastinal masses. Ninety-one cases (85.8%) were in stage III or IV at diagnosis. The 1-year and 5-year survival rates in patients with T-LBL/ALL were 36.1% and 8.1%, respectively. Patients older than 25 years and those presented in stage III or IV suggested a poor prognosis (P = 0.049 and 0.001, respectively). On the other hand, 29 of the 153 cases (19.0%) belonged to B-LBL/ALL. The median age of the patients was 14 years (ranged from 9 months to 75 years). The male-to-female ratio was 1.6:1 (including 18 males and 11 females). Seventeen patients (58.6%) presented with peripheral lymphadenopathy and 13 of them (44.8%) had involvement of bone marrow or peripheral blood. Mediastinal involvement was found only in 5 cases (17.2%). Twenty-one patients (72.4%) were in stage III or IV at diagnosis. The 1-year and 5-year survival rates were 53.3% and 36.7%, respectively. The remaining 11.7% cases (18/153 cases) were categorized as undefined type, with a negative staining for the following immuno-markers including: CD3ε/CD3, CD45RO, CD79a, CD20, MPO, CD5, CD56, cyclin D1, cytokeratin, neuron-specific enolase, chromogranin A and synaptophysin. The median age of the patients was 15.5 years (ranged from 4 to 53 years). The male-to-female ratio was 2.6:1 (including 13 males and 5 females). The percentage of T-LBL/ALL patients with mediastinal masses were significantly higher than that of B-LBL/ALL cases (P = 0.0003). There was no significant difference in prognostic parameters of T-LBL/ALL and B-LBL/ALL (P = 0.07). The difference in median survival time however was statistically significant (6.0 months +/- 1.1 months versus 15.0 months +/- 7.0 months).

CONCLUSIONSBoth TdT and CD99 are useful markers for the diagnosis of precursor lymphoblastic malignancy. T-LBL/ALL predominantly affects children or adolescent males and frequently presents with lymphadenopathy and mediastinal masses, whereas B-LBL/ALL are often accompanied by bone marrow and peripheral blood involvement. In general, T-LBL/ALL carries a poor prognosis. The prognostic criteria include age of older than 25 years and a classification of stage III or IV disease.

12E7 Antigen ; Adolescent ; Adult ; Age Factors ; Aged ; Antigens, CD ; metabolism ; Bone Marrow ; pathology ; Cell Adhesion Molecules ; metabolism ; Child ; Child, Preschool ; DNA Nucleotidylexotransferase ; metabolism ; Female ; Humans ; Immunophenotyping ; Male ; Middle Aged ; Neoplasm Staging ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; metabolism ; pathology ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; metabolism ; pathology ; Prognosis ; Retrospective Studies ; Survival Rate ; Young Adult

AIMTo investigate the neural network and cellular mechanisms of hippocampal epileptogenesis contralateral or ipsilateral to the side of acute tetanization (60 Hz, 2 s, 0.4 - 0.6 mA) of the posterior dorsal hippocampus (ATPDH).

METHODS10 trains of the ATPDH were administered into the CA1 basal dendritic region of the right hemisphere at an interval of 10 minutes.

RESULTS(1) The firing rate of CA1 single neuron in the right or the left hippocampus was inhibited respectively after the ATPDH, and the effects weakened gradually while the trains of the ATPDH increased. The inhibited firing rate and the transformed firing pattern from tonic one to clonic one were more obvious at the side contralateral to the stimulation (62.94% +/- 3.68%, 36.61% +/- 3.14%, P < 0.01). (2) Synchronous primary afterdischarges of depth EEG and single unit discharges were more commonly observed at the side ipsilateral to the ATPDH (P < 0.01). (3) Primary or secondary hippocampal network afterdischarges at high frequency were only found in CA1 region ipsilateral to the ATPDH. (4) Secondary afterdischarges of CA3 basal dendritic neural network were completely synchronized with those of subicular single neuron, which reoccurred and persisted several hours.

CONCLUSIONIt is possible that post-inhibition bursting of single neuron and recurrent network seizures in the hippocampus contralateral to the artificial focus be the important manifestation of the formation of "epileptic networks" across from one hemisphere to another.

Animals ; Electric Stimulation ; Hippocampus ; physiology ; Male ; Neural Pathways ; physiology ; Rats ; Rats, Sprague-Dawley ; Seizures ; etiology

OBJECTIVETo detect gene mutations of children with glucose-6-phosphorate dehydrogenase (G-6-PD) deficiency and of carriers of G-6-PD deficiency gene with the technique of polymerase chain reaction and denatured gradient gel electrophoresis (PCR-DGGE), and to explore the value of the technique in the diagnosis of G-6-PD deficiency and G-6-PD deficiency gene carrying.

METHODScDNAs were harvested by reverse transcription method after RNAs had been extracted from peripheral blood of 43 children with G-6-PD deficiency and of their family members (36 lineages). Electrophoresis behaviors of the fragment from exons 11-12 of G-6-PD cDNA were detected with the technique of PCR-DGGE. Gene sequencing was then performed for the abnormal electrophoresis bands.

RESULTSAbnormal electrophoresis bands were found in the 1304-1520 fragment of G-6-PD cDNA in 33 out of 36 family lineages. The G-6-PD/6-PGD ratio was below 1.00 in 9 mothers of patients. Three of them had the G-6-PD/6-PGD ratio lower than 0.50. The PCR-DGGE bands were the same in the 3 mothers. Gene sequencing showed double heterozygote in the 3 mothers, but the maternal carriers of G-6-PD deficiency gene who had normal G-6-PD/6-PGD ratio showed mono-heterozygote in gene sequencing. Three mutational sites were found in the 1304-1520 fragment, i.e., C1311TG1376T and G1388A. The electrophoresis behaviors were different among the 3 gene mutational sites.

CONCLUSIONSPCR-DGGE is a sensitive and reliable technique in the screening of gene mutations. It is useful in the diagnosis of G-6-PD deficiency, especially in the diagnosis of female G-6-PD deficiency gene carrying.

Base Sequence ; Electrophoresis, Polyacrylamide Gel ; Female ; Glucosephosphate Dehydrogenase Deficiency ; diagnosis ; genetics ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA

AIMTo observe the property alteration of K(v) channel in SLE patient's peripheral blood lymphocyte and its significant.

METHODSThe patch-clamp technique was used to record the current of K(V) channel in SLE patient's peripheral lymphocyte.

RESULTSThe current amplitude of K(V) channel in the SLE patient's lymphocytes decreased, it was (258.6 +/- 112.5) pA in healthy people, but in SLE patient it was (139.4 +/- 58.5) pA (P < 0.05). There was no other changes in the property of channel, include activation potential, inactivation property, channel closing kinetics and its pharmacological property.

CONCLUSIONThe decline of SLE patient's cell immunity may be related to the decrease of the amplitude of K(V) channel current.

Case-Control Studies ; Cell Membrane ; physiology ; Flow Cytometry ; Humans ; Lupus Erythematosus, Systemic ; blood ; pathology ; physiopathology ; Lymphocyte Count ; Lymphocytes ; physiology ; Monocytes ; metabolism ; physiology ; Patch-Clamp Techniques ; Potassium Channels, Voltage-Gated ; physiology

OBJECTIVETo investigate the influences of verapamil preconditioning on cardiac function in vitro and intracellular free Ca2+ and L-type calcium current (I(Ca-L)) in rat cardiomyocytes post ischemia-reperfusion (I/R) injury.

METHODSThe isolated rat hearts in control group (37 degrees C Tyrode solution perfusion for 30 min, n = 6), I/R group (no flow for 30 min followed 30 min reperfusion with 37 degrees C Tyrode solution, n = 7) and verapamil preconditioning group [37 degrees C Tyrode solution perfusion for 10 min, adding verapamil (20 micromol/L) to Tyrode solution and perfusion for another 30 min, followed then by 30 min no flow and 30 min reperfusion, n = 7] using Langendorff perfusion system. The fluorescence intensity of intracellular Ca2+ was detected with Fluo-3/AM loading by the laser scanning confocal microscope. The I(Ca-L) was recorded via whole-cell patch clamp technique in enzymatically dissociated single rat ventricular myocytes.

RESULTSAs expected, arrhythmias and cardiac dysfunction were shown post I/R injury. The fluorescence intensities of intracellular free Ca2+ in cardiomyocytes were significantly increased compared with control group (P < 0.01). By voltage clamp protocol, peak current densities of I(Ca-L) was significantly reduced and I-V curve significantly elevated. Post I/R injury compared with control group (P < 0.01) which could be reversed by Verapamil preconditioning. Verapamil preconditioning also significantly improved diastolic and systolic functions, and reduced the incidence of arrhythmias.

CONCLUSIONSMyocardial I/R injury might significantly impair heart functions and induce arrhythmias via cellular Ca2+ overload. Verapamil preconditioning could prevent heart I/R injury and reduce arrhythmias by decreasing influx of I(Ca-L), thereby stabilizing cardiomyocytes in myocardial stunning and avoiding occurrence of Ca2+-induced Ca2+ release during I/R injury.

Animals ; Calcium ; metabolism ; Calcium Channels, L-Type ; drug effects ; Ischemic Preconditioning, Myocardial ; methods ; Myocardial Reperfusion Injury ; metabolism ; physiopathology ; Myocytes, Cardiac ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Verapamil ; pharmacology

Objective To study the different effects of valsartan and extended realse nifedipine tablets on lowering blood pressure of essential hypertension patients and their reversal effects on left ventricular hypertrophy. Methods 100 cases of essential hypertension patients with left ventricular hypertrophy were randomly divided into valsartan group(group A) and adalt group(group B).Other antihypertensive drugs except diuretic were removed for 3 weeks.There were 50 cases in group A using valsartan 4～8mg qd,and 50 cases in group B using adalt 30～60 mg qd,the stud),lasted for 24 weeks.The blood pressure was measured and the altrasowic cardiogram examed in baseline and 24 weeks later.Results BP could be significantly reduced after treatment(P

Objective To establish a quantitative RT-PCR method with self-quenched fluorogenic probe for detection of bcr/abl mRNA in patients with chronic myeloid leukemia for providing a useful tool for diagnosis of CML,evaluation of therapeutic effect and monitoring of minimal residual disease(MRD). Methods bcr/abl gene from cultured K562 cells was amplified by conventional RT-PCR.The standard quantitative plasmid was constructed by A-T clone method.The self-quenched fluorogenic quantitative RT- PCR method(FQ-RT-PCR)for determination of bcr/abl mRNA was established successfully using the ABI PRISM 7000 PCR Detector.The linear range,sensitivity,stability,and repetitiveness of the method were determined.The marrow samples from 25 CML patients and 3 ALL patients were assessed.Results The sensitivity of the FQ-RT-PCR was 10 copies/?l recombined plasmid,and bcr/abl mRNA can be detected from 1 K562 cell in 10~5 normal cells.The linear range was 10~2-10~9 copies/?l recombined plasmid.The coefficient variation(CV)value was 2.1% in intra-assay and 6.1% in inter-assay.The median ber/abl mRNA expression level was 4.50?10~4 copies/?g RNA [(0.45-89.00)?10~4],5.45?10~4 copies/?g RNA [(2.95-19.30)?10~4 ],13.00?10~4 copies/?g RNA [(4.10-89.00)?10~4] and 2.35?10~4 copies/?g RNA [(0.45-5.12)?10~4] in 25 CML patients,11 patients in the incipient chronic phase,6 patients in blastic crisis,8 patients in chronic period after treatment,respectively.The bcr/abl mRNA level in blastic crisis was significantly higher than that in chronic phase(q= 3.41,P

Recombinant humanized anti-ricin monoclonal antibody (MIL50) is a recombinant humanized monoclonal antibody targeting ricin. In this study, an ELISA method was used to establish a method for the determination of MIL50 in macaque serum, and a cross design method was used. Twelve rhesus monkeys were intravenously injected 1 mg·kg^-1 test preparation (MIL50 freeze-died powder injection) and reference preparation (MIL50 liquid preparation) to determine the plasma concentration of MIL50 at different time points, and the pharmacokinetic parameters were analyzed to compare the pharmacokinetic characteristics of MIL50 liquid preparation and freeze-died powder injection in rhesus monkeys. Animal welfare and experimental procedures follow the regulations of the Animal Ethics Committee of the Chinese Academy of Medical Sciences and Use of Laboratory Animals and the regulations derived by the Animal Care and Welfare Committee of the Institute of Radiation Medicine, Academy of Military Medical Sciences (IACUC-DWZX-2020-503). The results showed that there was no significant difference between C_max and AUC_0-5d in the two groups. The liquid preparation was the reference preparation, with C_max ratio of 101.6% and AUC_0-5d ratio of 101.9%, the 90% confidence interval of C_max was 79.42%-129.92%, and the 90% confidence interval of AUC_0-5d was 85.72%-121.18%. These results suggested that different dosage forms of MIL50 had certain differences in the changes of blood drug concentration in rhesus monkeys.

OBJECTIVETo investigate the clinicopathological and immunohistochemical features of lymphoblastic lymphoma (LBL).

METHODSA retrospective clinicopathological study of 96 cases LBL was carried out. Immunohistochemical staining was used for the characterization and immunophenotyping.

RESULTSThe patients age ranged from 4 to 72 years, with a median of 16 years, 69 patients were male and 27 female. Seventy-three cases had superficial or multi-lymphoadenopathy and 31 of them had mediastinal masses. Bone marrow was involved in 15 cases. Seventy-three cases were in clinical stages III and IV. The median survival of the followed-up patients was 5.5 (2 approximately 120) months. TdT and CD99 positive reactions were 75.0% and 92.7%, respectively. Of the 96 cases, 78 displayed T-cell marker positivity and 18 B-cell markers. 82.1% of the patients younger than 30 years of age had significantly higher incidences of T-LBL (64 patients), and 93.6% of the patients with mediastinal masses expressed T-cell markers. The poor prognostic factors were T-cell tumors, clinical stages III and IV, Ki-67 PI < 80% and no chemotherapy (P < 0.01).

CONCLUSIONIn children and young males, mediastinal masses with superficial or multi-lymphoadenopathy favors the diagnosis of LBL, but negative TdT reaction can not exclude this diagnosis. T-LBL is more common than B-LBL. Clinical stages, immunophenotypes and the level of Ki-67 expression were closely related with prognosis of LBL.

Adolescent ; Adult ; Aged ; Antigens, CD20 ; analysis ; CD79 Antigens ; analysis ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Humans ; Immunohistochemistry ; Ki-67 Antigen ; analysis ; Leukocyte Common Antigens ; analysis ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; metabolism ; pathology ; Prognosis ; Survival Analysis ; Young Adult