Objective To evaluate the effects of alcohol extract of juglans mandshurica maxim (AEBJ) on immune function of ionizing irradiated mice.Methods The mice were randomly divided into normal control group,irradiating control group,low AEBJ(300 mg?kg-1)irradiating group,high AEBJ(600 mg? kg-1)irradiating group,low AEBJ plus drugs only group and high AEBJ plus drugs only group.The number of WBC and LYMPH,LYMPH%,viscera index,ability of lymphocytes transformation were examined.Results Compared with normal control group,the number of WBC and LYMPH,LYMPH%,viscera index,ability of lymphocytes transformation in irradiating control group were decreased significantly (P

Objective To study the effect on Henoch-Schnlein purpura nephritis with cyclophosphamide (CTX) intravenous pulse therapy.Methods Fourty cases of Henoch-Schonlein purpura nephritis were divided into observation group and control group.The observation group:8～10 mg/(kg?time) of CTX was added into 100 mL of saline which was dropped intravenously,2 days was a period of treatment, intermission of 30 d, and the total duration was 4-6 months. The control group only applied prednisone and dipyridamole.Results After CTX pulse therapy 24 h proteinuria determination and hematuria decreased obviously. The effect before and after therapy had significant difference (P

Objective: To observe the effect of acupuncture plus thunder-fire moxibustion on the expressions of matrix metalloproteinase-3 (MMP-3), tissue inhibitor of metalloproteinase-1 (TIMP-1) and transforming growth factor-β1 (TGF-β1) in cartilage of knee osteoarthritis (KOA) rats, and to explore the mechanism of acupuncture plus thunder-fire moxibustion in the treatment of KOA. Methods:Thirty Sprague-Dawley (SD) rats were randomly divided into a blank control group, a model group and an acupuncture-moxibustion group by random digits table, 10 rats in each group. Rats in the model group and the acupuncture-moxibustion group were injected with papain in the right posterior knee joint to prepare the models. The levels of MMP-3 and TIMP-1 in rat synovium of each group were measured by enzyme-linked immunosorbent assay (ELISA) after 2 weeks of treatment. The level of TGF-β1 was determined by Motic B5 Micro-camera system. Results:The levels of MMP-3 and TIMP-1 in the cartilage of the model group were significantly higher than those in the blank control group (allP<0.01); the levels of MMP-3 and TIMP-1 in the acupuncture-moxibustion group were lower than those in the model group, and the between-group differences were statistically significant (all P<0.05). The levels of MMP-3 and TIMP-1 in the acupuncture-moxibustion group were higher than those in the blank control group, and the differences were statistically significant (all P<0.05). The level of TGF-β1 in cartilage tissues of the model group was significantly lower than that in the blank control group (P<0.01); the level of TGF-β1 in the acupuncture-moxibustion group was higher than that in the model group (P<0.05), but it was lower than that in the blank control group, and the between-group difference was statistically significant (P<0.05). Conclusion: Acupuncture plus thunder-fire moxibustion can effectively recover the abnormal expressions of MMP-3 and TIMP-1 in KOA model rats and somewhat up-regulate TGF-β1, which may be one of its mechanisms of acupuncture plus thunder-fire for KOA.

Objective To study application of Chinese general practice assessment questionnaire (GPAQ)consultation version 2.0 in evaluating patients' satisfaction with community health-care service (CHS).Methods First,origihal GPAQ consultation version 2.0 in English was translated into Chinese language by three experts in general practice/family medicine field,then translated it back into English,finally a Chinese version of GPAQ was formed.Ten medical doctors who were on duty on 25 April,2008 at each out-patient clinic from CHS of Daxing,Nancaiyuan,Yongning and Hancunhe were selected randomly as targets for evaluation of service satisfaction.A sample of the first 30 patients(1200 in total)aged more than 16 years who visited the clinic on that day were selected to fill out the Chinese GPAQ consultation version 2.0 to express their satisfaction with the medical doctors they visited,and reliability and validity of the questionnaire was then evaluated.Results Cronbach's alpha ranged 0.734 to 0.813 for each domain of the Chinese version of GPAQ2.0.and cumulated contribution of three common factors with the eigen-value more than one reached 58.722%.Scores of patients' satisfaction withreceptionists,access,continuity,doctor's communication skills andpatient enablement were significantly lower,as compared to those of national bench-mark in the United Kingdom,with t-values of -6.397,-11.729,-6.328,-59.871 and -2.210,and all P＜0.05.respectively.Conclusions The Chinese version of GPAO 2.0 achieves good reliability and modcrate validity,and can be used as an instrument for evaluating patient's satisfaction with quality of CHS in China as further revision in the future studies.

Human leukemia is closely associated with various genetic alterations such as chromosomal translocations and gene mutations. The use of retroviral transduction/bone marrow transplantation mouse model harboring these genetic abnormalities has been critical in understanding the molecular pathogenesis of leukemia and exploring new therapeutic target. Additional genetic events are verified to cooperate with fusion genes resulting from chromosomal translocations in acute myeloid leukemia (AML) to develop a leukemic phenotype in mice, such as C-KIT N822K with AML1-ETO, FLT3-ITD with PML-RARα, Meis1 with NUP98-HOX, and Cdx4 with MLL-AF9. Mouse model shows that BCR/ABL fusion gene induces chronic myeloid leukemia (CML), and suggests that GATA-2 L359V and high expression of Hes1 are key molecules in acute myeloid transformation of CML. Furthermore, combination therapy with Imatinib and arsenic sulfide for CML mice exerts more profound therapeutic effects than either drug as a single agent. This review focuses the recent progress and application of retroviral-mediated mouse models of myeloid leukemia, and discusses some factors influencing the mouse model establishment, including retroviral construction, retrovirus titer and hematopoietic microenvironment.
Animals ; Disease Models, Animal ; Leukemia, Myeloid ; genetics ; Mice ; Retroviridae ; genetics

Lysine decarboxylase is a key enzyme involved in the upstream biosynthesis of lycopodium alkaloids (LAs) such as huperzine A, contributing to the decarboxylation of lysine to 1,5-pentanediamine (cadaverine). Three lysine decarboxylase genes (HsLDC-L1, HsLDC-L2, HsLDC-L3) were successfully cloned from Huperzia serrata using transcriptomic sequence data mining strategy combined with reverse transcription PCR. The physicochemical properties, secondary and tertiary structures, amino acid identities, and evolutionary relationship of the three LDCs were analyzed by online bioinformatics analysis platforms and DNAMAN, MEGA 7.0 software, revealing that all of these proteins had the conserved PLP binding domain and active site residues were completely conserved in LDCs. Phylogenetic analysis showed that these LDCs were located in the same branch as other known LDCs from LA-producing plants. Accordingly, the ORFs of these three HsLDCs were inserted into different expression plasmids for further expression in E. coli. However, only HsLDC-L1 was successfully expressed in E. coli BL21 (DE3) by inserting into a pCold TF vector. The recombinant protein was purified by Ni²⁺ affinity chromatography purification. HsLDC-L1 contains 469 amino acid residues, with a calculated molecular weight of 50.50 kDa. HsLDC-L1 expectedly catalyzed the decarboxylation of lysine to produce cadaverine. In addition, HsLDC-L1 can also catalyze the generation of putrescine from ornithine. However, it cannot catalyze the decarboxylation of tyrosine, phenylalanine, tryptophan and histidine. The results not only provide insight into the biosynthesis of LAs including huperzine A, but also provide a critical genetic element for the overproduction of Δ¹-piperideine and pelletierine, the essential biosynthetic precursors of LAs, using synthetic biology strategies.

OBJECTIVETo characterize the structural and the functional feature of a novel gene HSPCSET isolated from human CD34+ hematopoietic stem/progenitor cells (HS/PCs).

METHODSBioinformatic technology was used to identify the structural features of the HSPCSET protein and perform the multiple sequence alignment. Yeast-two-hybrid system was used to identify the proteins interacting with the HSPCSET protein. After sequencing, we selected out the positive clones which had clear functions, and carried out beta-gal experiment and GST pull down assay to confirm the results. The cellular location of the HSPCSET was checked by immunofluorescence assay.

RESULTSThe HSPCSET protein belongs to a SET domain family, which is evolutionarily conserved across species. It implied that HSPCSET may have biologically important function. Using yeast-two-hybrid system, we showed that the protein sequence with SET domain might bind to 13 proteins, which involved in signaling transduction, transcriptional regulation, apoptosis, tumorigenesis, development, etc. And 4 proteins (GADD34, SIVA, DNAJ and PHF1) were confirmed by one-on-one back of the hybrid experiment, beta-gal test and GST pull down assay. When GADD34 and HSPCSET were co-transfected, they co-localized in the nucleus, suggesting a strong interaction.

CONCLUSIONThe novel gene HSPCSET is likely to have biologically important function. This study provides the basis for further studies of its function in hematopoiesis and tumorigenesis.

Amino Acid Sequence ; Animals ; Antigens, Differentiation ; metabolism ; Cell Cycle Proteins ; metabolism ; Computational Biology ; Conserved Sequence ; Hematopoietic Stem Cells ; metabolism ; Humans ; Molecular Sequence Data ; Protein Phosphatase 1 ; Protein Structure, Tertiary ; Proteins ; chemistry ; genetics ; metabolism ; Sequence Homology, Amino Acid ; Two-Hybrid System Techniques

Abnormalities of chromosome 11 involving mixed lineage leukemia (MLL) on 11q23 are often seen in acute myeloid leukemia (AML)-M5 or AML-M4. The fusion gene of MLL-PTD and MLL plays a critical role in the pathogenesis of these AML. However, rare chromosome abnormalities have been identified in this type of leukemia. To explore whether there were other MLL gene mutations at M4 and M5, in this study all of the MLL exons were sequenced at cDNA level. 25 patients with de novo AML-M4 or M5 with normal karyotypes excluding M4eo and MLL fusion gene or MLL-PTD were selected, the amplification and direct sequencing analysis of full length MLL gene exons were carried out, then the mutations found were verified at genomic DNA level. Furthermore, the point mutations were tested in normal samples and a larger group of AML patients using the platform of Mass Array. The results showed that high-frequency deletion/insertion and point mutations in RD, PHD, TAD and SET domains of MLL were found, while these alterations in normal samples and other subtypes of AML samples were also verified, and without significant difference (P > 0.05). It is concluded that a variety of deletions/insertions in MLL mRNA and point mutations are respectively alternative splicing of MLL gene at transcriptional level and single nucleotide polymorphism. These alternations together constituted genetic polymorphisms of MLL. Although these variations may not play a direct role in the molecular pathogenesis of AML-M4 or M5, their correlations to clinical treatment and prognosis need to be further explored.
Alternative Splicing ; Base Sequence ; Chromosomes, Human, Pair 11 ; genetics ; DNA Mutational Analysis ; Histone-Lysine N-Methyltransferase ; Humans ; Leukemia, Monocytic, Acute ; genetics ; Leukemia, Myelomonocytic, Acute ; genetics ; Molecular Sequence Data ; Mutation ; Myeloid-Lymphoid Leukemia Protein ; genetics ; Oncogene Proteins, Fusion ; genetics

Objective To investigate the characteristics and diagnostic value of 18F-fluorodeoxyglucose (FDG) PET/CT in patients with secondary malignant peripheral nerve lesions. Methods 18F-FDG PET/CT studies of 8 cases of secondary malignant peripheral nerve lesions confirmed by histopathology or follow-up were analyzed retrospectively. The maximum standardized uptake value ( SUVmax ) of infiltrating peripheral nerves and contralateral normal peripheral nerves was measured and compared with their morphological appearances on CT. Paired student t-test was performed by SPSS 10.0. Results Twelve secondary malignant peripheral nerve lesions with high 18F-FDG metabolism were found in 8 cases. On PET imaging,the lesions distributed along the neurovascular tissues or intervertebral foramina with appearances resembling those of fibre bundles,radices or nodes on PET but no density differences with the surrounding soft tissue or fat planes on CT. The SUVmax was 6.86 ± 3.87. The contralateral normal peripheral nerves showed no abnormal 18F-FDG uptake with a SUVmax of 1.10 ±0.46,which was significantly different from that of the secondary malignant peripheral nerve lesions (t = 9.231,P ＜ 0.001 ). Conclusion 18 F-FDG PET/CT may be useful in locating the secondary malignant peripheral nerve lesions and in assessing its regional infiltration.

To investigate the potential role and the mechanism of PLZF-RARalpha/RARalpha-PLZF double fusion gene in the pathogenesis of acute promyelocytic leukemia (APL) in vivo at systematic biological level, PLZF-RARalpha/RARalpha-PLZF double transgenic mouse model was established by intercross; the integration and expression of fusion genes were analyzed by PCR and RT-PCR; the disease phenotype was detected by morphological and pathological examination of peripheral blood and bone marrow cells, as well as flow cytometry assays; the effects of ATRA with or without tricostatin A on bone marrow blast cells from PLZF-RARalpha/RARalpha-PLZF double TM were observed. The results showed that leukemia occurred in 5 PLZF-RARalpha/RARalpha-PLZF double TM 7, 7, 9, 11 and 11 months respectively, out of them two (40%) with classic APL features, the others (60%) with chronic myeloid leukemia through an observation period of 18 months. The leukemia occurrence of PLZF-RARalpha/RARalpha-PLZF TM was about 10%, which was similar to PLZF-RARalpha TM as that reported before. The latency was over 6 months, not earlier than PLZF-RARalpha TM only. No morphologic changes of PLZF-RARalpha/RARalpha-PLZF double TM blast cells to ATRA were observed, but increased cytoplasmic-nuclear ratio and nuclear condensation in bone marrow blast cells were found in combination of ATRA with tricostatin A. It is concluded that PLZF-RARalpha/RARalpha-PLZF double fusion gene transgenic mice have heterogeneity of pathogenesis. HDAC inhibitors such as trichostatin A, in combination with ATRA, induce differentiation of the blast/promyelocytic cells from PLZF-RARa/RARa-PLZF double TM, but not ATRA alone.
Animals ; Antigens, CD34 ; blood ; Bone Marrow Cells ; drug effects ; immunology ; pathology ; Cell Differentiation ; drug effects ; Chorionic Gonadotropin ; genetics ; Disease Models, Animal ; Female ; Flow Cytometry ; Humans ; Hydroxamic Acids ; pharmacology ; Leukemia, Promyelocytic, Acute ; blood ; genetics ; pathology ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Inbred CBA ; Mice, Transgenic ; Oncogene Proteins, Fusion ; genetics ; Pedigree ; Receptors, Chemokine ; blood ; Tretinoin ; pharmacology