We investigated and analyzed the first case of human infection with avian influenza A(H9N2) virus in Yunnan Province,China,so as to provide a better basis for preventing and controlling human infections with viruses of animal origin in the future.We carried out the field epidemiological survey among the patient,close contacts and the live poultry markets,detected and analyzed the samples from patient and the outdoor environment.Results showed that the 9-month-old boy was a case of human infection with avian influenza A(H9N2) virus with the history of live poultry markets exposure and the results of nucleic acid detection and virus isolation.There was a lot of contamination of the avian influenza virus in the live poultry markets.The second generation cases have not occurred.The monitoring of pneumonia of unknown etiology and influenza like cases in medical institutions is the important means to find timely cases of human infected with avian influenza.Regular disinfection and closing-down of live poultry markets are key measures to reduce the exposure opportunity.

Although children with differentiated thyroid cancer ( DTC) are in a high proportion of cure, due to the absence of the standard management of the evaluation, treatment and follow-up of children with thyroid neoplasia, the aggressive therapy including total thyroidectomy followed by radioactive iodine ( RAI) ablation with iodine-131 ( 131 I) could not be ignored. For the majority of children, total or near-total thyroidectomy is recommended. Central neck dissection is recommended for children with malignant cytology and clinical evidence of gross extrathyroidal invasion and/or loco-regional metastasis on pre-operative. Selective use of 131 I in children with DTC was based on pre-and post-operative staging. Multi-disciplinary collaborative management is needed to improve their outcomes and to minimize adverse effects as far as possible.

Objective To solve the agglomeration problem in the former process, dry granulation technology was used to prepare the granules of Lianhua Qingwen Capsules. Methods Complexity and granule yield coefficient were set as inspection indexes. The optimum subsidiary material and its amount were optimized. The parameters of dry granulation technology were optimized by orthogonal test. Then, granule yield, angle of repose, and bulk density were compared with those of wet granulation technology. Results Starch was set as subsidiary material. The optimum technology is roll pressure of 12 MPa, rotation speed of 5 r/min, and feed speed of 10 r/min. The yield of granules prepared by dry granulation technology was significantly higher than that of wet granulation technology. Conclusion The dry granulation technology can effectively improve the agglomeration of the granulation process of Lianhua Qingwen Capsules, and was suitable for granulating process of Lianhua Qingwen Capsules.

Objective To evaluate the relationship between the level of AMP-activated protein kinase (AMPK) in hippocampus and cognitive dysfunction after splenectomy in aged rats.Methods Sixty-three male aged Sprague-Dawley rats,weighing 480-550 g,were randomly divided into 3 groups (n =21 each):control group (group C),anesthesia group (group A),and surgery group (group S).Morris water maze (MWM) test was performed to evaluate the spatial learning and memory ability before surgery and at 1,3 and 7 days after surgery.The escape latency and swimming distance were recorded.Seven rats were chosen after MWM test was performed at 1,3 and 7 days after surgery and sacrificed.Their hippocampi were removed for detection of the expression of AMPK and phosphorylated (p-AMPK) (by Western blot).Results Compared with the baseline,the escape latency and swimming distance were significantly prolonged at 1 and 3 days after surgery in group S (P ＜ 0.05).Compared with group C,the escape latency and swimming distance were significantly prolonged at 1 and 3 days after surgery,the expression of AMPK was up-regulated at 1,3 and 7 days after surgery,and the expression of p-AMPK was up-regulated at 1 and 3 days after surgery in group S (P ＜ 0.05),and no significant changes were found in the indices mentioned above in group A (P ＞ 0.05).Conclusion Increased AMPK level in hippocampus is the regulatory mechanism of the body adapting to the development of cognitive dysfunction after splenectomy in aged rats.

Objective To investigate the expression and prognostic value of Ikaros6 in adult T-cell acute lymphoblastic leukemia(T-ALL) patients.Methods The RNA was extracted from mononuclear cells of bone marrow in 74 adult T-ALL patients.The expression of Ikaros6 was examined by reverse transcription-PCR,and the results were confirmed by sequencing.Clinical features and prognosis were analyzed based on clinical data.Results In 74 T-ALL patients,the incidence rate of Ikaros6 was 21.6 ％ (16/74).Extramedullary infiltrations were occurred often (x2 =4.084,P =0.043),had higher WBC (103.15 × 109/L vs 15.60×109/L,t =0.214,P =0.831),more severe anemia (75.95 g/L vs 107.00 g/L,t =1.504,P =0.142) and lower platelet count (26.0×109/L vs 67.0×109/L,t =1.421,P =0.164) in patients with Ikaros6 positive.Meanwhile Ikaros6-positive patients had inferior survival and were increased risk of relapse as compared with the Ikaros6-negative patients.Conclusions The incidence of Ikaros6 is high in adult T-ALL patients.Ikaros6-positive patients are more likely to have extramedullary infiltration and higher WBC,meanwhile they had inferior survival and increased risk of relapse.Thus Ikaros6 may be served as a valuable factor for risk stratification and prognosis evaluation in adult T-ALL patients.

Objective To investigate the expression and clinical significance of P-selectin (CD62P) in bone marrow hematopoietic stem cells of patients with acute leukemia (AL). Methods The CD62P expression in bone marrow mononuclear cells of 15 healthy donors and 56 untreated patients with AL, were examined by flow cytometry. Results The average rate of CD62P expression was (6.72±7.64) % in hematopoietic stem cells (CD+45 CD+34 CD-38) of the 38 patients with acute myeloid leukemia (AML), was (3.46±2.51) % in hematopoietic stem cells (CD+45 CD+34 CD+19) of the 12 patients with B-acute lymphoblastic leukemia (B-ALL), and was (6.23±4.95) % in hematopoietic stem cells (CD+45 CD+34 CD+7) of 6 patients with T-acute lymphoblastic leukemia (T-ALL). The expression rates in those AL patients were higher than that in the healthy controls (1.04 ±1.23) % (t = 2.847, 3.284, 3.091, respectively, P ＜0.01), while there was no difference between the control group and the group who reach CR after routine treatment (t =1.932, P ＞0.05). Furthermore, the leukocyte,hemoglobin and platelet count in CD+62P patients with AML and T-ALL were significantly higher than CD-62P ones (t =4.153, 8.095, 8.289, 7.235, 8.692, 9.832, respectively, P ＜0.05), but there was no significant difference between CD+62P and CD-62P patients with B-ALL (t =0.340, 1.142, 0.019, respectively, P ＞0.05).Conclusion The CD62P is one of the markers of platelet activation, and its expression varies in different types of AL. The CD62P in hematopoietic stem cells of AL could be regarded as a new sign for the leukemic stem cells, as well as a helpful prognostic indicator in treatment response assessment.

Multiplex reverse transcription-polymerase chain reaction (M-RT-PCR) has been proved to possess great clinical potential for simultaneous screening of 29 chromosomal translocations in acute leukemia. To evaluate the clinical value of M-RT-PCR in hematologic malignancies, bone marrow samples from 90 patients with various hematologic malignancies, including 25 acute myelogenous leukemia (AML), 22 acute lymphoblastic leukemia (ALL), 27 chronic myelogenous leukemia (CML), 4 myeloproliferative diseases (MPD), 3 chronic lymphoblastic leukemia (CLL), 3 non-Hodgkin's lymphoma (NHL), 3 myelodysplastic syndrome (MDS), 2 multiple myeloma (MM) and 1 malignant histocytosis (MH) were subjected to both M-RT-PCR and chromosome karyotypic analysis. Some of cases were subjected to follow-up examination of M-RT-PCR during the period of clinical complete remission (CR) for detection of minimal residual leukemia. In our hand, 12 of 29 chromosomal translocation transcripts including TEL/PDGFR, DEK/CAN, MLL/AF6, AML1/ETO, MLL/AF9, BCR/ABL, MLL/MLL, PML/RARu, TLS/ERG, E2A/HLF, EVI1 and HOXI1 were detected in 57 cases (63.3 %) of the 90 samples, which were in consistence with the results of karyotypic analysis. Furthermore, M-RT-PCR had also shown good clinical relevance when used as an approach to detect minimal residual leukemia. We concluded that M-RT-PCR could be used as an efficient and fast diagnostic tool not only in the initial diagnosis of hematologic malignancies but also in subsequent monitor of minimal residual leukemia.

The effects of two different histone deacetylase (HDAC) inhibitors, sodium butyrate (NaB) and trichostatin A (TSA),on apoptosis of human leukemic cells in vitro and the molecular mechanisms were investigated. The experiments were divided up 5 groups: control group, NaB group, TSA group, NaB+Z-VAD-FMK group and TSA+Z-VAD-FMK group. The apoptosis rate was determined by morphological analysis and flow cytomytry. The expression of Daxx, Bcl-2, and Bcl-xl proteins was detected by Western blot. NaB and TSA could induce the apoptosis of HL-60 and K562 cells, and Z-VAD-FMK caused a marked decrease in apoptosis induced by HDAC inhibitors. HDAC inhibitors could down-regulate the expression of Daxx protein, but had no significant influence on the expression of Bcl-2 and Bcl-xl proteins. The results suggested that NaB and TSA induce distinct caspase-dependent apoptosis of human leukemic cells through down-regulating the expression of Daxx protein in vitro.

Nowadays the role of genetic findings in determining the diagnosis, therapy and prognosis of acute myeloid leukemia (AML) has become more valuable. To improve and validate the detection of clonal chromosomal aberrations in leukemia, we designed a combined application of karyotyping with multiplex reverse transcription-polymerase chain reaction (RT-PCR) and fluorescence in situ hybridization (FISH), and addressed the expression and distribution of fusion genes among the subtypes of Chinese adult patients with de novo AML. Multiplex RT-PCR assays were performed on 477 samples from newly diagnosed AML patients, and cytogenetic data were obtained from 373 of them by R or G banding techniques and those in some cases were confirmed by FISH. The PCR products in some suspected cases were tested by two-directional sequencing. The results showed that except unqualified samples, fusion genes were detected by multiplex RT-PCR in 211 of 474 patients (44.51%), including AML1-ETO, CBFβ-MYH11, PML-RARα, PLZF-RARα, NPM-RARα, MLL rearrangements, BCR-ABL, DEK-CAN, SET-CAN, TEL-PDGFR, TLS-ERG, AML1-MDS1 (EVI-1). In 373 patients, who took both multiplex RT-PCR and karyotype analysis, the detection rate of chromosomal aberrations by using multiplex RT-PCR and karyotyping was 160/373 (42.89%) and 179/373 (47.98%) respectively, and the combination could optimize the detection rate of clonal genetic abnormalities to 216/373 (57.90%). The PCR results from 11 cases "normal" in karyotyping but abnormal in RT-PCR for MLL rearrangements were confirmed by two-directional sequencing. It is concluded that karyotype studies remain the cornerstone for genetic testing; conventional cytogenetics and molecular-based methods are complementary tests for the detection of clonal genetic aberrations in AML, especially for the cryptic or submicroscopic aberrations. Once a genetic marker has been identified by combined analysis, it could be used to monitor residual disease during/after chemotherapy, by quantitative RT-PCR and/or FISH.