In order to study the regularity of shedding virus from infected SPF chickens and the formation of aerosol of H9N2 subtype AIV, SPF chickens were bred in a positive and negative pressure isolator. Aerosol samples were collected by AGI-30 (All Glass Impinger-30) extractor, and simultaneously trachea and cloaca samples were collected by tracheal swabs and cloacal swabs in different periods after challenged with vi- ruses. The above-mentioned samples were detected by HI, Dot-ELISA and RT-PCR methods. The results in- dicated that aerosols were isolated from the 4 days to the 43 days after inoculation. It was proved that H9N2 subtype AIV could copy themselves in respiratory passage and cloaca, and then could formation of aerosols. AIV H9N2 subtype could be isolated from cloacal and tracheal swabs 3 days after inoculation and lasted for 45 days, viruses were detected from all infected SPF chickens on 7 days.

AIMTo design and synthesize some cephalotaxine and drupacine derivatives with different substituentes on C3'-N of taxol side chain.

METHODSProtective side chain acid VI (4'S,5'R) was prepared from optically active (2'R,3'S) methyl beta-phenyl glycidate I in five steps. The desired acids were coupled with cephlotaxine and drupacine respectively in the presence of 2-DPC/DMAP, followed by acidic hydrolysis and acylating to give novel alkloid esters with different substitutes on C3'-N.

RESULTSThe seven new esters were studied for antitumor activity, the results showed that the antitumor activity was influenced by the substituentes on C3'-N.

CONCLUSIONIt might provide some rational basis for further structral modification.

Antineoplastic Agents, Phytogenic ; chemical synthesis ; chemistry ; pharmacology ; Cell Line, Tumor ; Colonic Neoplasms ; pathology ; Esters ; Harringtonines ; chemical synthesis ; chemistry ; pharmacology ; Humans ; KB Cells ; Liver Neoplasms ; pathology ; Molecular Structure

OBJECTIVETo investigate the effects of GnRH analogues GnRHa and GnRHant on the MAPK pathway in rat Leydig cells.

METHODSRat Leydig cells were primarily cultured for 24 hours in vitro and serum-starved for 2 hours, followed by treatment with GnRHa (10(-7) mol/L) or GnRHant (10(-6) mol/L) for 0, 5, 15, 30, 60 and 90 minutes, with the 0 min group as the control. Then the protein levels of phosphorylated ERK (p-ERK) and phosphorylated p38 (p-p38) were detected by Western blot, and that of p-ERK determined by the same means after co-incubation of GnRHa or GnRHant with the PKC inhibitor GF109203X at 1, 5, 10 and 20 micromol/L.

RESULTSAfter stimulation of the Leydig cells with GnRHa or GnRHant for different times, the protein level of p-p38 showed no significant difference from that of the control group (P > 0.05). Then the Leydig cells were treated with GF109203X at different concentrations for 20 minutes and with addition of GnRHa for another 10 minutes. The level of p-ERK was significantly decreased (P < 0.05) by GF109203X at 10 and 20 micromol/L. Compared with the control, the p-ERK expression was increased by 65% at 15 minutes (P < 0.05) in the GnRHant stimulation group, by 81% (to the peak) at 30 minutes (P < 0.05), began to fall at 60 minutes, and returned to the base level at 90 minutes. The p-ERK level exhibited no significant difference from that of the control (P > 0.05) after treatment of the Leydig cells with different concentrations of GF109203X for 20 minutes and then with GnRHant for 30 minutes.

CONCLUSIONThe ERK MAPK activation induced by GnRHa depends on the PKC pathway, but not that induced by GnRHant. The p-38 MAPK pathway may not be involved in the effect of GnRH analogues on rat Leydig cells.

Animals ; Cells, Cultured ; Gonadotropin-Releasing Hormone ; analogs & derivatives ; pharmacology ; Leydig Cells ; drug effects ; metabolism ; MAP Kinase Signaling System ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley

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

BACKGROUNDPerforator flaps are used extensively in repairing soft tissue defects. Superior gluteal artery perforator flaps are used for repairing sacral defects, but the tension required for direct closure of the donor area after harvesting of relatively large flaps carries a risk of postoperative dehiscence. This research was to investigate a modified superior gluteal artery perforator flap for repairing sacrococcygeal soft tissue defects.

METHODSFrom June 2003 to April 2010, we used our newly designed superior gluteal artery perforator flap for repair of sacrococcygeal soft tissue defects in 10 patients (study group). The wound and donor areas were measured, and the flaps were designed accordingly. Wound healing was assessed over a follow-up period of 6 - 38 months. From January 1998 to February 2003, twelve patients with sacrococcygeal pressure sores were treated with traditional methods, VY advancement flaps or oblong flaps, as control group.

RESULTSAfter debridement, the soft tissue defects ranged from 12 cm × 10 cm to 26 cm × 22 cm (mean 16.3 cm × 13.5 cm). Four patients were treated using right-sided flaps ranging from 15 cm × 11 cm to 25 cm × 20 cm (mean 18.2 cm × 14 cm). Four patients were treated using left-sided flaps, and two were treated using both right- and left-sided flaps. Suction drains were removed on postoperative Days 3 - 21 (mean 5.9) and sutures were removed on postoperative Days 12 - 14. Each flap included 1 - 2 perforators for each of the donor and recipient sites. Donor sites were closed directly. All flaps survived. In eight patients, the wounds healed after single-stage surgery. After further debridement, the wounds of the remaining two patients were considered healed on postoperative Days 26 and 33, respectively. The rate of first intention in the study group (80%, 8/10) significantly increased than that of control group ((25%, 3/12), χ(2) = 4.583, P = 0.032). Follow-up examinations found that the flaps had a soft texture without ulceration. In the two patients without paraplegia, the range of motion of the hip joints was not affected.

CONCLUSIONThe use of the quadrilobed superior gluteal artery perforator flap can overcome the disadvantages of traditional perforator flaps and represents an improved approach for repairing soft tissue defects in the sacrococcygeal region.

Adult ; Debridement ; Female ; Humans ; Male ; Middle Aged ; Perforator Flap ; Sacrococcygeal Region ; surgery ; Soft Tissue Injuries ; surgery ; Wound Healing

OBJECTIVE: To review the clinical data of pathological morphology, diagnosis, surgical treatment of cor triatriatum in 15 patients. METHODS: Fifteen patients with a mean age of (14.6+/-10.3) years (range from 6 months to 40 years) were performed operations under extracorporeal circulation. Fourteen of the patients had cor triatriatum sinister, and 1 had cor triatriatum dexter; 12 of the 15 patients had other cardiac abnormalities. The excision of the fibromuscular membrane was accomplished through a right atrial incision in all of the 14 cases, and the associated abnormalities were corrected at the same time. RESULTS: One patient died after the operation, and the other survivors had good outcome. CONCLUSION Operation is necessary if the diagnosis is clear. The patients generally have good prognosis. Surgical results of cor triatriatum depend on the complexity of associated defects and the adequacy of the repair.
Adolescent ; Adult ; Child ; Child, Preschool ; Cor Triatriatum ; surgery ; Female ; Humans ; Infant ; Male ; Retrospective Studies ; Young Adult

Objective To investigate the southern region of Zhejiang hepatitis E virus (HEV) infection. Methods A cluster sampling strategy was used to sample all blood donors from February to Ocwber in 2008 in Wenzhou blood center. Their blood was tested for IgG and lgM antibody against HEV. Reverse transcriptase-polymerase chain reaction(RT-PCR) and sequencing were applied to detect its genotypo and sequence homology in HEV IgM-positive specimen. Results The prevalence of anti-HEV IgG in 3044 cases of blood donors was 33.28%. IgG increased with age. There are certain increase in positive rates between the 20-year-old group and over 40 years of age group from 21.16% to 50.36%. The positive rate of IgM was 0.92% . The ratio of infection among different age group was the highest in the age range from 31to 40 years and up to 1.90%. lgG and IgM through their negative and positive analysis of samples found in their group with donors age, sex and blood type does not significantly related to each other. Nucleic acids were found in three cases through PCR amplification in all 28 cases of HEV IgM positive samples.The total positive rate was one-thousandth, of which two cases for gene 4,1 cases of infection for gene 1. Conclusion The results indicate that there was a certain percentage of HEV virus in voluntary blood donors in south Zhejiang.

BACKGROUNDDiabetes mellitus is associated with coronary dysfunction, contributing to a 2- to 4-fold increase in the risk of coronary heart diseases. The mechanisms by which diabetes induces vasculopathy involve endothelial-dependent and -independent vascular dysfunction in both type 1 and type 2 diabetes mellitus. The purpose of this study is to determine the role of vascular large conductance Ca(2+)-activated K(+) (BK) channel activities in coronary dysfunction in streptozotocin-induced diabetic rats.

METHODSUsing videomicroscopy, immunoblotting, fluorescent assay and patch clamp techniques, we investigated the coronary BK channel activities and BK channel-mediated coronary vasoreactivity in streptozotocin-induced diabetic rats.

RESULTSBK currents (defined as the iberiotoxin-sensitive K(+) component) contribute (65 ± 4)% of the total K(+) currents in freshly isolated coronary smooth muscle cells and > 50% of the contraction of the inner diameter of coronary arteries from normal rats. However, BK current density is remarkably reduced in coronary smooth muscle cells of streptozotocin-induced diabetic rats, leading to an increase in coronary artery tension. BK channel activity in response to free Ca(2+) is impaired in diabetic rats. Moreover, cytoplasmic application of DHS-1 (a specific BK channel b(1) subunit activator) robustly enhanced the open probability of BK channels in coronary smooth muscle cells of normal rats. In diabetic rats, the DHS-1 effect was diminished in the presence of 200 nmol/L Ca(2+) and was significantly attenuated in the presence of high free calcium concentration, i.e., 1 mmol/L Ca(2+). Immunoblotting experiments confirmed that there was a 2-fold decrease in BK-b(1) protein expression in diabetic vessels, without altering the BK channel α-subunit expression. Although the cytosolic Ca(2+) concentration of coronary arterial smooth muscle cells was increased from (103 ± 23) nmol/L (n = 5) of control rats to (193 ± 22) nmol/L (n = 6, P < 0.05) of STZ-induced diabetic rats, reduced BK-b(1) expression made these channels less sensitive to intracellular Ca(2+), which in turn led to enhanced smooth muscle contraction.

CONCLUSIONSOur results indicated that BK channels are the key determinant of coronary arterial tone. Impaired BK channel function in diabetes mellitus is associated with down-regulation of BK-b(1) expression and reduction of the b(1)-mediated BK channel activation in diabetic vessels.

Animals ; Blotting, Western ; Coronary Vessels ; metabolism ; Diabetes Mellitus, Experimental ; metabolism ; physiopathology ; Diabetes Mellitus, Type 1 ; metabolism ; physiopathology ; Electrophysiology ; Large-Conductance Calcium-Activated Potassium Channels ; metabolism ; Male ; Muscle, Smooth, Vascular ; metabolism ; Rats ; Rats, Sprague-Dawley