The cytolytic effect of perforin is a mechanism of anti-virus,killing microbial-infected cells and tumor cells.Perforin is a very important non-specific immune factors in fish.In order to understand the function of perforin,the cDNA of grass carp perforin C-terminal peptide was amplified from grass carp liver and kidney cDNA library.It contains a protein kinase C conserved region 2(C2).The cDNA was connected with pET32a,and transformed to expression bacteria DE3.PFP-C was expressed by a prokaryotic expression system and then purified by affinity chromatography.It showed a significant haemolytic activity when tested with rabbit red cells,the optimal pH for haemolytic activity was 7.5,and its haemolytic function dependents on Ca2+ apparently.

Adolescent ; Adult ; Antigens, Nuclear ; metabolism ; Brain Diseases ; complications ; diagnosis ; pathology ; surgery ; Brain Neoplasms ; pathology ; Child ; Child, Preschool ; Diagnosis, Differential ; Epilepsy ; etiology ; Female ; Ganglioglioma ; pathology ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Infant ; Magnetic Resonance Imaging ; Male ; Malformations of Cortical Development ; classification ; complications ; diagnosis ; pathology ; surgery ; Malformations of Cortical Development, Group I ; Microtubule-Associated Proteins ; metabolism ; Neoplasms, Neuroepithelial ; pathology ; Nerve Tissue Proteins ; metabolism ; Neurofilament Proteins ; metabolism ; Retrospective Studies ; Vimentin ; metabolism ; Young Adult

OBJECTIVETo investigate the effect of hepatocyte growth factor (HGF) on graft-versus-host disease (GVHD) and graft-versus-leukemia (GVL) after allogeneic bone marrow transplantation (allo-BMT) and related mechanism in acute lymphoblastic leukemia (ALL) mice.

METHODSTwenty nude mice were randomly divided into control (group A) and test (group B) groups for monitoring relapse, and 20 BALB/c mice into control (group C) and test (group D) groups for GVHD. HGF as injected from day 0 to day 7 after BMT for groups B and D, while PBS for A and C. CD4(+) and CD8(+) T cell were evaluated by flow cytometry. The survival of mice after BMT was recorded. The level of tumor necrosis factor-alpha (TNF-alpha) was evaluated by ELISA.

RESULTSThe median past-BMT survival were 7.00 +/- 1.58, 9.00 +/- 1.58, 11.00 +/- 3.95 and 24.00 +/- 13.44 days for groups A, B, C, D, respectively, being prolonged in group D. HGF could decrease the quantity of CD4(+) T cells [group D (10.39 +/- 1.15)% vs group C (13.50 +/- 1.80)%, P < 0.01] and increase CD8(+) T cell [group D (12.25 +/- 2.85)% vs group C (6.12 +/- 1.99)%, P < 0.01], decrease the level of TNF-alpha in transplanted ALL mice [group D (112.10 +/- 18.99) pg/ml vs group C (143.90 +/- 25.35) pg/ml, P < 0.01] and reduce the degree of GVHD.

CONCLUSIONHGF could alleviate post-allo-BMT GVHD but retain GVL effect.

Animals ; Bone Marrow Transplantation ; Disease Models, Animal ; Female ; Graft vs Host Disease ; prevention & control ; Graft vs Leukemia Effect ; drug effects ; Hepatocyte Growth Factor ; pharmacology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; immunology ; surgery ; Random Allocation ; Transplantation, Homologous

OBJECTIVETo detect plasma concentrations of vascular endothelial cell growth factor (VEGF) and tissue factor (TF) in children with acute lymphoblastic leukemia (ALL) and explore their clinical significance in ALL.

METHODSThirty-three children with newly diagnosed ALL, including 18 cases of low risk, 7 cases of moderate risk and 8 cases of high risk, were enrolled in this study. Twenty-five patients received a complete remission and 8 cases were in non-remission after conventional remission induction chemotherapy. Plasma concentrations of VEGF and TF in the patients were detected using ELISA before and after treatment. Sixteen healthy children served as normal control group.

RESULTSPlasma concentrations of VEGF and TF in ALL patients before treatment were significantly higher than those in normal controls (P < 0.01). Plasma concentrations of VEGF and TF in the non-remission group before treatment were significantly higher than those in the remission group (P < 0.05) and the control group (P < 0.01). After treatment the plasma concentrations of VEGF and TF in the non-remission group were not significantly reduced and higher than those in the remission and the control groups (P < 0.01). There were significant differences in plasma concentrations of VEGF and TF among the low-risk, moderate-risk and high-risk groups before and after treatment (P < 0.05). Plasma concentrations of VEGF and TF in the high risk group were not significantly reduced after treatment and higher than those in the control group (P < 0.01). A linear correlation was noted between plasma VEGF and TF concentrations in ALL patients before treatment (r=0.50, P < 0.01).

CONCLUSIONSVEGF and TF play an important role in the development of ALL and may be useful to the evaluation of the severity and the outcome in ALL.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; blood ; etiology ; Thromboplastin ; analysis ; Vascular Endothelial Growth Factor A ; blood

To observe the effects of hepatocyte growth factor (HGF) on graft-versus-host disease (GVHD) and Th1/Th2 related cytokines in mice with acute lymphoblastic leukemia (ALL) after allogenic bone marrow transplantation (allo-BMT), BALB/c mice were conditioned by total body irradiation with 11 Gy and then were transplanted with allogeneic bone marrow after establishing ALL model. BALB/c mice were divided into groups A and B. The mice of group A were injected subcutaneously with HGF from day 0 to 7 after allo-BMT, and the mice of group B were injected subcutaneously with PBS from day 0 to 7 after allo-BMT. The symptoms of GVHD and the GVHD pathological changes of liver and small intestine and skin were observed. The serum levels of both IFN-gamma and IL-4 were determined by ELISA. The results showed that the score of GVHD in group A was lower than that in group B (P < 0.05). The levels of IFN-gamma in both groups A and B were all higher than that in normal group (P < 0.05 and P < 0.001, respectively), However, the level of IFN-gamma in group A was lower than that in group B (P < 0.01). The levels of IL-4 in both group A and B were all lower than that in normal group (P < 0.05), but the level of IL-4 in group A was higher than that in group B (P < 0.05). It is concluded that HGF can alleviates the severity of GVHD, because of its balancing the Th1/Th2-related cytokines after allo-BMT.
Animals ; Bone Marrow Transplantation ; adverse effects ; methods ; Cytokines ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Graft vs Host Disease ; immunology ; prevention & control ; Hepatocyte Growth Factor ; pharmacology ; Interferon-gamma ; blood ; Interleukin-4 ; blood ; Male ; Mice ; Mice, Inbred BALB C ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; blood ; immunology ; surgery ; Th1 Cells ; immunology ; Th2 Cells ; immunology ; Transplantation, Homologous

PURPOSE: Modafinil is a wake-promoting agent that has been proposed to improve cognitive performance at the preclinical and clinical levels. Since there is insufficient evidence for modafinil to be regarded as a cognitive enhancer, the aim of this study was to investigate the effects of chronic modafinil administration on behavioral learning in healthy adult rats. METHODS: Y-maze training was used to assess learning performance, and the whole-cell patch clamp technique was used to assess synaptic transmission in pyramidal neurons of the hippocampal CA1 region of rats. RESULTS: Intraperitoneal administration of modafinil at 200 mg/kg or 300 mg/kg significantly improved learning performance. Furthermore, perfusion with 1mM modafinil enhanced the frequency and amplitude of spontaneous postsynaptic currents and spontaneous excitatory postsynaptic currents in CA1 pyramidal neurons in hippocampal slices. However, the frequency and amplitude of spontaneous inhibitory postsynaptic currents in CA1 pyramidal neurons were inhibited by treatment with 1mM modafinil. CONCLUSIONS: These results indicate that modafinil improves learning and memory in rats possibly by enhancing glutamatergic excitatory synaptic transmission and inhibiting GABAergic (gamma-aminobutyric acid-ergic) inhibitory synaptic transmission.
Adult ; Animals ; CA1 Region, Hippocampal ; Excitatory Postsynaptic Potentials ; Humans ; Inhibitory Postsynaptic Potentials ; Learning* ; Memory ; Neurons ; Perfusion ; Rats* ; Synaptic Potentials ; Synaptic Transmission*

OBJECTIVETo evaluate the whole body MRI and diffusion-weighted MRI in detecting intranodal lesions in patients with lymphoma.

METHODSWhole body MRI and diffusion-weighted MRI (DWI) were performed in 23 patients with histologically proven lymphoma. A conventional coronal MRI scan from head to inguinal groove was done for whole body scanning. In the DWI, axial MRI scans were performed after segmentation based on SENSE technique, and all images were merged into whole body image reconstruction by software.

RESULTS417 lymph nodes were detected by MRI in the 23 patients. The overall positive rate of whole body MRI and DWI was 79.1% and 89.7%, respectively. It was 70.9% versus 85.2% and 79.4% versus 90.1% for the lymph nodes of < 2 cm and 2-3 cm in diameter, with a significant difference between the two methods (P < 0.01). However, it was 94.7% versus 97.9% for the lymph nodes of > 3 cm in diameter, not significantly different between the two methods (P > 0.05). Both methods had similar sensitivity in detecting the lymph nodes in the neck, supraclavicular and infraclavicular fossae, mediastinum and axillary fossa. However, the positive rate of whole body MRI was 51.2%, 43.8% and 52.2%, significantly less sensitive than 83.7%, 71.9% and 87.0%, respectively, by DWI in detecting the lymph nodes in the retroperitoneal space, pelvic cavity and inguinal groove (all P < 0.01).

CONCLUSIONBoth whole body MRI and diffusion-weighted MRI have a relative high sensitivity in detecting intranodal lesions for patients with lymphoma, showing a certain value in clinical application.

Adolescent ; Adult ; Aged ; Diffusion Magnetic Resonance Imaging ; methods ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphoma ; diagnosis ; pathology ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Neoplasm Staging ; Whole Body Imaging ; methods ; Young Adult

OBJECTIVETo summarize the clinical experience in repair of deep burn and traumatic wounds with combined transplantation of different types of pedicled skin flaps in lower extremities.

METHODSTwo hundred and thirty-six patients with 271 deep wounds in lower extremities after burn or trauma were repaired with muscular skin flaps, local fascial flaps and island flaps with vascular pedicle (more than 20 types) in our department from Jan. 1998 to Sept. 2008.

RESULTSComplete necrosis of skin flaps occurred in 1 case, congestion and necrosis over the edge of skin flaps occurred in 3 cases, which were healed after grafting, and other skin flaps survived well with soft texture. Skin flaps were too bulky in 26 cases, among them 17 cases were thinned, and the appearance of other skin flaps were satisfactory. In 68 patients with functional region injury were recovered to certain extent without contracture.

CONCLUSIONSSkin flaps with pedicles, multiple transplantations if necessary, can repair deep wounds satisfactorily in lower extremities after deep burn or trauma injury.

Adolescent ; Adult ; Aged ; Burns ; surgery ; Buttocks ; surgery ; Child ; Child, Preschool ; Female ; Humans ; Lower Extremity ; injuries ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; Skin Transplantation ; Surgical Flaps ; Wound Healing ; Young Adult

OBJECTIVETo investigate the validity of transesophageal echo-Doppler technique for hemodynamic monitoring of patients with extensive burns during shock stage to provide guidance for fluid resuscitation.

METHODSMongrel dogs were inflicted with 50% TBSA full-thickness burns. Cardiac output (CO) were determined by Swan-Ganz catheter, and other hemodynamic indices by transesophageal echo-Doppler device, including aortic blood flow (ABF), stroke volume (SVa), left ventricle ejection time (LVET), peak velocity (PV), max acceleration (max ACC), total systemic vascular resistance (TSVR) were collected before and after burns. Hemodynamic indices of ten patients with extensive burns were collected by transesophageal echo-Doppler device with in 24 hours after the injury.

RESULTSBoth CO and ABF showed continuous decrease after burns, and there was positive correlation between them (r = 0.90, P < 0.01). The myocardiac contractibility (ACC, PV) and the preload of the left ventricle (LVET) were also decreased continuously after bums,but the postburn afterload (TSVR) were obviously increased. For the patients with conventional therapy, the hemodynamic state of low cardiac output and high peripheral resistance during early period was converted to high cardiac output and low peripheral resistance 24 hours after injury.

CONCLUSIONTransesophageal echo-Doppler technique is useful in monitoring hemodynamic in patients with extensive burn, and it is non-invasive, real-time, synchronized and rapid, and it can provide complete profile of hemodynamic data.

Animals ; Blood Flow Velocity ; Burns ; diagnostic imaging ; therapy ; Dogs ; Fluid Therapy ; Humans ; Male ; Ultrasonography, Doppler ; Vascular Resistance

Objective To study the HIV drug resistance (HIVDR) transmission in Kunming city of Yunnan province in 2010.Methods Referring to the guidelines for HIV drug resistance threshold survey (HIVDR-TS) set by WHO,62 plasma samples of recently reported HIV-infected individuals who were older than 25 years of age,were collected from January to August 2010.Genotyping of pol genetic mutations associated with HIVDR with reverse transcriptional PCR was performed and the prevalence of HIV-1 drug resistance transmission was evaluated.Results Of the 62 plasma samples,54 were successfully sequenced and genotyped on pol sequence.Based on the pol sequences,HIV subtypes including CRF08_BC ( 53.2％ ),CRF07_BC ( 25.5％ ),CRF01_AE ( 19.1％ )and C (2.1％) were identified.According to the time of sampling,the first 47 sequenced samples were used for drug resistance prevalence analysis.A protease inhibitor (PI) relative mutation was found in one sample.Based on the WHO standard,the prevalence of transmitted HIV-1 drug resistance was ＜5％.Conclusion HIV-1 drug resistant strains transmission was still catalogued as low prevalence level in Kunming.To prevent the increase of HIVDR prevalence,normative treatment and scientific management to AIDS patients seemed to be quite important.