Animals ; Drugs, Chinese Herbal ; chemistry ; Humans ; Medicine, Chinese Traditional ; Microscopy, Confocal ; Muscle, Smooth, Vascular ; ultrastructure

PBL tutoring is quite different from the traditional tutoring and emphasizes the importance of student-centred instead of teacher-centred education.In PBL,teachers guide students to find answers to their own questions and facilitate students' learning process.Therefore,the tutor role is complex and has a fundamental change.

Objective To know the expression of 47 kD protein of orientia tsutsugamushi(Ot) Karp strain and test its immunogenicity and antigenicity.Methods Forty-seven kD protein gene that code the whole mat peptide was amplified from the genomic DNA of Ot.Karp strain by PCR and constructed to a recombinant plasmid pGEX-47.The E. coli cells BL21 were transformed with pGEX-47,then the transformants were induced to express the recombinant 47 kD protein by IPTG. The antigenicity and immunogeni-(city) were identified by Dot Blot method and the experiment of immunizing mice respectively.Results 1. The PCR product of 47 kD protein gene of Ot.Karp strain was obtained and its sequence was the same as the published 47 kD protein gene.2. The E.coli expression plasmid pGEX-47 constructed was identified containing the 47 kD protein gene fragment by restrication analysis and PCR identification,and an expression band about 47 kD was detected by SDS-PAGE of the purified product from GST-fusson protein expressed by pGEX-47. 3.Dot Blot analysis and the experiment of immunizing mice testified its antigenicity and immunogenicity.Conclusions The 47 kD protein gene of Ot Karp strain is amplified that code the whole mat peptide, which can be expressed in prokaryotic cells and the expressed protein has antigenicity and immunogenicity.

Anemia, Hemolytic, Congenital ; etiology ; Female ; Hepatitis ; complications ; Humans ; Infant ; Syndrome

Objective To study the effects of CD2-associated protein (CD2AP) on podocyte adhesion and extension ability and to explore its possible mechanism. Methods Conditionally immortalized murine podocyte cell line was cultured in RPMI 1640 medium at 33℃permissive conditions. The podocytes were transfected with CD2AP small interfering RNA (siRNA) and serambing sequences labeled with fluorescein were taken as control. The transfected podocytes were trypsinized and seed into collagen IV coated plates. The relative cell adhesion and cell area were examined 90 min later. Apoptotic rates of CD2AP siRNA transfected podoeytes and different PAN concentrations incubated podoeytes were detected by flow cytometer. The distribution of F-actin was observed under laser scanning confoeal microscope. Nephrin protein expression and its phosphorylation level were examined by immunofluorescence and Western blot. Results The relative ceil adhesion of CD2AP siRNA transfected podocytes was apparently lower than that of control group[(41.72±6.07)% vs (64.46±8.53)%, P<0.05]. The cell area analysis had the similar result. The apoptotic rate of CD2AP siRNA transfected podocytes was significantly higher than that of the controls [(5.73±0.61)% vs (3.26±0.45)%, P<0.05]. 100 mg/L PAN could markedly induce podocytes to apoptosis and impair cell adhesion ability (P<0.05). Nevertheless, no significant difference was found in cell body spreading (P>0.05). The distribution of F-actin in CD2AP depletion podocytes was apparently altered. The expression of nephrin protein and its phosphorylation level was conspicuously descended to some degree (P<0.05). Conclusions CD2AP depletion facilitates podocyte apoptosis and impairs cell adhesion function. Cytoskeleton confusion and nephrin signaling weakness caused by CD2AP depletion may he partly responsible for the decline of cell adhesion and spreading.

Objective: To investigate the alterations of bcl-2 gene family in the rat brain and the molecular mechanism of neuronal apoptosis follow ing traumatic brain injury (TBI). Methods: Male Sprague-Dawley rats were subjected to lateral fluid percussion brain injury(FPBI) of moderate severity. bax and bcl-xL mRNA and protein expression was detected by RT-PCR an d immunohistochemistry. In addition to morphological evidence of apoptosis, TUNE L histochemistry was used to identify DNA fragmentation in situ under both l ight and electron microscope, whereas characteristic internucleosomal DN A fragm entation of apoptosis was demonstrated by DNA gel electrophoresis. Resul ts: bcl-xL mRNA and protein decreased in the ipsilateral hemisphere t o the impact site as early as 6 h post-injury［(67.42±7.54)% and (85.85±5.72)% r espectively］. The decrease in bcl-xL mRNA and protein preceded apoptosis was observed 12 h post-injury. And this was the main cause of up-regulation of the ratio of bax to bcl-xL in the acute period(minutes-hours) followin g FPBI. bax mRNA and protein were observed to rise slowly, doubled 3 d post- injury, returned to sham level slowly. The delayed cell death (days-weeks) migh t associated with the up-regulation of pro-apoptotic gene bax. Conclusio n: The expression of bcl-xL and bax coincide with apoptosis following TBI. The reg ulation of bax and bcl-xL by TBI occur before transcription. The balance of bax/bcl-xL ratio determines the neurocytes to survive or die following FPBI.

Objective: To investigate the alteration of bcl- 2 gene family in the rat brain and the molecular mechanism of neuronal apoptosis following traumatic brain injury. Methods: Male Sprague -Dawley rats were subjected to lateral fluid percussion brain injury(FPI) of mo derate severity. Bcl-2, Bcl-x and Bax protein expression was detected by immun ohistochemistry. Results: (1) The immunoreactivity of Bcl-2 and Bcl-x protein decreased in the hippocampus ipsilateral impact site as early as 6 h post-injury, and this was the main cause of down-regulation of the ratio of Bcl-2+Bcl-x to Bax. (2) During 1-3 d after injury, the Bax protein express i on increased significantly, while the Bcl-2 and Bcl-x protein expression decre ased relatively slow. The decreased ratio of Bcl-2+Bcl-x to Bax was mainly due to the Bax up-regulation. Conclusion: The bcl-2 gene family is involved in neuronal apoptosis after FBI, and the protein expression alteration of the family members leads the neuronal cell to apoptosis.

Inservice Training ; Occupational Health Services ; organization & administration

Background and purpose:The patients with aggressive lymphoma who have a poor prognosis and unlikely to be cured with conventional chemotherapy. This study was aimed to evaluate the effect of high-dose etoposide in mobilization followed auto-SCT in treating refractory lymphoma. Methods:40 patients [median age 33 (13-61) years] with refractory non-Hodgkin’s lymphoma (NHL, n=32) or Hodgkin’s lymphoma (HD, n=8) received high-dose etoposide [VP16 10-15 mg/(kg·d)×2 d] in mobilization in our center. Remission status prior to mobilization was PD (n=40). The use of such granulocyte colony-stimulating factor [G-CSF, 5-10μg/(kg·d)] mobilized peripheral blood stem cells (PBSC) after high-dose etoposide until the end of leukapheresis. Peripheral blood stem cell was collected and frozen in-80℃refrigerator. All these patients received auto peripheral blood stem cell transplantation (auto-PBSCT). Conditioning regimen was BEAM (n=19, 47.5%) or CBV (n=21, 52.5%). Results:Twenty-eight pa-tients (70%) were assessable for response after high-dose etoposide at a median pretreatment time of 39 days (range 17-172 days), 12 patients (30%) had no response. Median follow-up of 28 (4-66) months, 16 patients (40%) reached CR after auto-PBSCT. Fifteen of the 28 patients (53.6%) who had response to high-dose etoposide reached CR, 4 patients (14.3%) reached PR, 9 patients (32.1%) succumb to progression of disease. One of the 12 patients (8.3%) who had no response to high-dose etoposide reached CR, 1 patients (8.3%) reached PR, 10 patients (83.4%) succumb to progression of disease. The estimated 1-year OS and EFS were 69%and 56.7%respectively, 2-years OS and EFS were 63%and 52%respectively. The prognosis of the patients who had no response to etoposide was poor. The estimated 1-year OS and EFS were 25%and 16.7%respectively. Two group of comparison differences have statistics signiifcance (P<0.01). Conclusion: High-dose etoposide could be used in refractory lymphoma as rescue therapy in mobilization. It can increase the EFS and OS of patients who had response. The hematopoietic stem cells collection and hematopoietic reconstitution are not affected by etoposide.

Objective: To investigate the alteration of bcl 2 gene family in the rat brain and the molecular mechanism of neuronal apoptosis following traumatic brain injury. Methods: Male Sprague Dawley rats were subjected to lateral fluid percussion brain injury(FPI) of moderate severity. Bcl 2, Bcl x and Bax protein expression was detected by immunohistochemistry. Results: (1) The immunoreactivity of Bcl 2 and Bcl x protein decreased in the hippocampus ipsilateral impact site as early as 6 h post injury, and this was the main cause of down regulation of the ratio of Bcl 2+Bcl x to Bax. (2) During 1 3 d after injury, the Bax protein expression increased significantly, while the Bcl 2 and Bcl x protein expression decreased relatively slow. The decreased ratio of Bcl 2+Bcl x to Bax was mainly due to the Bax up regulation. Conclusion: The bcl 2 gene family is involved in neuronal apoptosis after FBI, and the protein expression alteration of the family members leads the neuronal cell to apoptosis.