Lentiviral vector mediated up?regulation of miR?126 promotes migration and invasion on AGS cells / 实用医学杂志

Objective To establish an AGS cell line that stably expressing miR?126 and to study the effect of miR?126 on the proliferation and metastatic abilities of the AGS cell line in vitro. Methods AGS cells were infected by lentivirus with Lv?has?mir?126. After confirmation by RT?PCR ,CCK?8 and clone formation assays were used to evaluate the effect of miR?126 on AGS cell growth. Transwell migration and invasion assays were used to evaluate the effect of miR?126 on metastasis of AGS cells. Results We verified correct construction of recombinant AGS cells. RT?PCR confirmed mRNA levels of miR?126 existed significantly differences among the recombinant cell lines (P< 0.05). Proliferation assays and clone formation assays did not show a remarkable growth suppression in AGS?mir?126 cell line. However,transwell assay showed a notable acceleration in AGS?mir?126(P< 0.05). Conclusions We successfully constructed recombinant AGS cell line with stably high miR?126 expression level. MiR?126 could facilitate the metastasis of AGS cell in vitro.

Construction and practice of a novel teaching pattern for laboratory diagnostics based on organ-system diseases / 中华检验医学杂志

Clinical medicine and its teaching patterns put forward the new demand to the teaching of laboratory diagnostics. According to the teaching objectives and development tendency of laboratory diagnostics, a novel teaching pattern based on organ-system diseases was established through systematical reform measures. The course content system was reconstructed and focused on the diseases. And the professional teacher group carefully orchestrated, applied new teaching methods,such as case-based learning and problem-based learning. The independent learning on a resource sharing network platform was encouraged, and the evaluation system was innovated. The novel teaching pattern has obtained gratifying achievement, and showed a bright prospect of development.

Application of virtual simulation technology to laboratory medicine / 中华检验医学杂志

Medical virtual simulation teaching environment is an advanced and complex combination of virtual simulation technology and medical technology training .As an advanced information technology , it has been attracting the attention of medical education researchers .Based on the characteristics of medical laboratory technology education , the virtual simulation experimental teaching center was established and a framework of the virtual simulation training environment was designed in the Department of Laboratory Medicine, Southern Medical University .Virtual simulation softwares and tool softwares were put into use widely in our experimental teaching .As it turns out, this simulated training environment has many merits such as risk-free safety , great efficiency , genuine learning experience and wide application potential , which partly overcomes the restriction of the teaching process brought by the condition of the practical operation and improves the teaching effect significantly .

Role of glycogen synthase kinase 3β in maturation and function of murine myeloid dendritic cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the role of glycogen synthase kinase 3β (GSK-3β) in the maturation and function of murine bone marrow-derived dendritic cells (BMDCs).</p><p><b>METHODS</b>Mature DCs (mDCs) induced by LPS were examined for GSK-3β phosphorylation level with Western blotting before and after LPS exposure. To explore the role of GSK-3β in maturation and function of DCs, we added SB216763, a selective inhibitor of GSK-3β, in the cell culture of immature DCs (iDCs), and examined CD40 and CD86 expressions in the cells by flow cytometry and the expression of IL-6, IL-12 and IL-10 mRNA by real-time PCR; the changes of the immunogenicity of the cells was evaluated by mixed lymphocyte reaction. The expression of GSK-3β and RelB was examined by Western blotting in DC2.4 cells transfected with a lentiviral vector over-expressing murine GSK-3β gene.</p><p><b>RESULTS</b>LPS exposure significantly lowered GSK-3β activity in iDCs as demonstrated by increased Ser9 phosphorylation and reduced Tyr216 phosphorylation. GSK-3β inhibition induced DC maturation by increasing the expression of surface costimulatory molecules CD40 and CD86, lowered the expressions of IL-6 and IL-12 while enhanced the expression of IL-10 in iDCs, and impaired mixed lymphocyte reaction of the cells. In DC2.4 cells, lentivirus-mediated over-expression of GSK-3β obviously down-regulated the expression of RelB.</p><p><b>CONCLUSIONS</b>GSK-3β is a crucial enzyme involved in the differentiation and maintenance of an immature phenotype of DCs. GSK-3β is constitutively active in iDCs to inhibit their spontaneous maturation. DCs become phenotypically mature after inhibition of GSK-3β, which also executes a proinflammatory task in DC activation. The reduction of RelB protein levels as a result of GSK-3β overexpression supports GSK-3β as a new target for inducing tolerogenic DCs.</p>

Plasma levels of mannan-binding lectin-associated serine protease 2 in children with upper respiratory tract infection / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the significance of plasma levels of mannan-binding lectin (MBL)-associated serine protease 2 (MASP2) in children with upper respiratory tract infection (URTI).</p><p><b>METHODS</b>A total of 103 children with URTI and 35 healthy children were examined for plasma levels of MASP2 and C-reactive protein (CRP). According to CRP levels, white blood cell count (WBC), stage of infection, and administration of treatments, the children with URTI were divided into the elevated CRP group (n=48) and the normal CRP group (n=54), elevated WBC group (n=61) and normal WBC group (n=40), the early stage of infection without treatment group (n=68) and mid-late stage of infection with treatment group (n=35).</p><p><b>RESULTS</b>Plasma MASP2 levels was significantly higher in URTI group than in the healthy control group (P<0.001) and showed a close correlation with age (r=0.302, P<0.01). Plasma MASP2 level was significantly correlated with CRP level in elevated CRP group (r=0.310, P<0.05) but not in normal CRP group (P>0.05), correlated with WBC in elevated WBC group (r=0.392, P<0.01) but not in normal WBC group (P>0.05), and was significantly higher in early stage infection without treatment group than in mid-late stage of infection with treatment group (P<0.01). MASP2, MBL2 and CRP genes had a common binding site for the transcription factor HNF-4α.</p><p><b>CONCLUSIONS</b>MASP2 may be an acute-phase protein, and its plasma level might serve as a new reference index in the diagnosis of URTI in children.</p>

Construction and practice of the innovation talents training mode for laboratory medicine / 中华检验医学杂志

Based on the developing trend of laboratory medicine and the internal rules of discipline construction,series of research and reform practices in the curriculum system were put forward,including practice teaching,innovation education and instructional technology for laboratory medicine.An innovation talents training mode for laboratory medicine is gradually formed and innovative education is pushed on systematically,effectively and abidingly.The new mode has made periodical results and good prospects in cultivating students with innovative quality and ability.

Exploration and practice of constructing a characteristic specialty for laboratory medicine / 中华医学教育探索杂志

According to the internal subject discipline of laboratory medicine and requests for national characteristics specialties,the Laboratory Medicine's construction can be strength-ened in educational object,teaching crew,curriculum system,teaching condition,cultivating quality and other aspects to strive for provincial and even national characteristics specialty.

Clinical significance of serial monitoring of co-stimulating signals in double hand allograft / 中国组织工程研究

BACKGROUND: Researches on the hand allograft have been transited from laboratory aspects to clinical application.OBJECTIVE: To evaluate the clinical significance of serial monitoring of co-stimulating signals in double hand transplantation.DESIGN: Case controlled observation.SETTING: Department of Traumatic Orthopaedics, Nanfang Hospital,Southern Medical University.PARTICIPANTS: From September 2001 to December 2001, a patient who was proposed to have double hand transplantation in the Department of Traumatic Orthopaedics, Nanfang Hospital Affiliated to Southern Medical University was taken as the subject of the experiment, meanwhile 15healthy adult man were enrolled as controls.METHODS: Before operation, blood sample was collected from the patient once before given immunodepressant and twice after given immunodepressant; blood sample was also collected before arteriovenous connection during operation, as well as once a day during the first week after operation, once every other day on the second week, twice a week on the third and fourth week, followed by once a week for total two month. The first week after operation was taken as inducing period with the remainder as maintaining period. 2 mL peripheral venous blood was collected from healthy adult and anti-coagulated by EDTA of mass scores of 20 g/L.Flowcytometry was used to carry out a serial monitoring of T cell surface co-stimulating moleculars (CD28, CD54, CD11a).MAIN OUTCOME MEASURES: The change of co-stimulatingmolecules on the surface of T lymphocyte before and after transplantation.RESULTS:The changes of the three co-stimulating molecules were consistent with each other after double hand transplantation (CD28,CD54,CD11a),displaying decrement during inducing period comparing to pre-operation [(9.84±5.28)%, (55.50±3.62)%; (71.03±5.33)%, (95.10±1.26)%;(9.40±9.17)%, (29.70±3.23)%] and keeping at lower level at maintaining period [ (22.54±6.56) %, (91.28±8.12 ) %, ( 11.22±4.08 ) %].CONCLUSION:The serial monitoring of the co-stimulating signals in the peripheral blood of patients with linb allograft is beneficial to the observation of the posopertaive immunologic reaction, providing important guidance for the application of immunosuppression medicine, as well as the prediction and diagnosis of postoperative immune rejection. Meanwhile, costimulating signal induced immunological tolerance is proved promising for the application of hand transplantation.

Early immunologic status of patients following hand allotransplantation / 中国组织工程研究

BACKGROUND: There are a lot of immunologic studies about limb allotransplantation in animal experiment. But, it is only early investigation in clinic; its clinical immunologic study needs further accumulation.OBJECTIVE: To dynamically analyze the early immunologic state change in patients following hand allotransplantation.DESIGN: Controlled trial.SETTING: Department of Orthopaedics, Guangzhou General Hospital,Guangzhou Military Area Command of Chinese PLA; Department of Traumatic Orthopaedics and Department of Laboratory Medicine, Nanfang Hospital, First Military Medical University of Chinese PLA.PARTICIPANTS: Two patients who underwent unilateral hand allotransplantation in the Department of Orthopaedics, Guangzhou General Hospital,Guangzhou Military Area Command of Chinese PLA were enrolled, serving as experimental group. The observation was between September 1999 and March 2000. Twenty persons, including 12 male and 8 female, who homochronously received health examination, aged 20 to 45 years, were enrolled, serving as healthy control group. They all had no reactive immune and infectious diseases, and voluntarily participated in the trial.METHODS: Peripheral blood was collected from 2 patients who underwent hand allotransplantation once respectively at pre-operative 1 day and 3 days. Blood collecting was performed once per day at post-operative 1 week, three times per week at post-operative 2 to 4 weeks, twice per week at 5 to 8 weeks post-operation, once per week at 9 to 16 weeks post-operation, twice per month at 5 to 6 months post-operation. ① Peripheral blood T cell subgroups (CD3+,CD4+,CD8+T cells)were detected by flow cytometer,serum panel reactive antibody (PRA) by ELISA method, serum C-reactive protein by turbidimetric immanoassay (TIA), serum creatine kinase (CK) by enzyme dynamics method. ②Mixed lymphocyte reaction(MLR): mitomycin C-treated donor peripheral blood lymphocytes were used as stimulator, and proliferative reaction of peripheral blood lymphocyte of patients to donor transplanted antigen was detected with the incorporation of 3H-TDR method (Negative: There was no significant difference between the mean value of stimulation index and 1, conversely positive). Autogenic peripheral blood lymphocytes treated with the same way replaced donor stimulator, serving as control. Stimulation index of each specimen was calculated (Stimulation index=Experiment cmp/controlcpm), serving as control index. Peripheral blood T-cell subgroups (CD3+,CD4+,CD8+T cell), serum PRA, C-reactive protein and CK were detected in 20 persons in healthy control group;Twenty persons were randomly divided into 10 groups. Two persons in each group were used as donor and recipient mutually and performed MLR.MAIN OUTCOME MEASURES: ① Peripheral blood T cell subgrpups (CD3+,CD4+,CD8+T cell). ②PRA. ③ C-reactive protein. ④CK. ⑤MLR.RESULTS: ①CD3+,CD4+,CD8+T cell levels were obviously decreased within one week after operation. CD3+ and CD4+T cell levels both recovered to be the pre-operative levels, but CD8+ level exceeded pre-operative level significantly [CD3+: (66.43±4.56); CD4+: (30.55±3.94); CD8 +:(33.45 ±2.69)]. There was no significant difference between experiment group and control group. ②Serum PRA was 0 to 10%, there was no significant difference as compared with control group. ③ Serum C-reactive protein was 0 to 0.359 mg/L, there was no significant difference as compared with control group. ④ Serum CK was 25 to 170 mmol/L, and there was no significant difference as compared with control group. ⑤ MLR after transplantation was negative, and it turned into be positive 5 months later.They were all positive in control group.CONCLUSION: Short-term change and long-term redistribution of T cell subgroups are closely related to immunosuppressive agent, suggesting that immunosuppressive agent has obvious effect on T-cell subgroup following hand allotransplantation. Immuno-induction schedule make patients be in immune suppression state, which effectively avoid early rejection. But patients cannot bear specificity yet; they need the inhibition of immunosuppressive agents.

The practice and contents of pre-job training of laboratory medicine undergraduate / 中华医学教育探索杂志

Clinical practice is an important stage in clinical education and the future career of the graduates. Taking the students of Clinical Inspection Major as an example,this article analyzes the questions need to be paid attention to in the clinical practice and the importance of pre-post training in increasing the quality of clinical practice.

Exploration and practice on the model curricula development of clinical immunology examination / 中华医学教育探索杂志

The construction of the model curricula is the important branch of teaching quality engineering. The key to the development of the model curricula lies in the reform of the teaching model. As required to the national standards of the model curricula and to the purpose of innovative educational philosophy,with the years of continuous exploration and practice,the clinical immunology examination curriculum has kept developing from the teaching staff,teaching condition,content of course,teaching method and means etc.,which has brought about satisfactory teaching effects. Thus it is forming the model curricula with special features.

Dynamic detection of T cell subsets and CD3＋HLA-DR＋T cells in peripheral blood with hand allograft recipients / 临床检验杂志

Objective To study dynamic changes of T cell subsets and CD3＋HLA-DR＋T cells in peripheral blood of composite tissue transplantation-hand allograft recipients. Methods The levels of CD3＋、CD3＋CD4＋、CD3＋CD8＋、CD3＋HLA-DR＋T cells and ratio of CD4/CD8 in peripheral blood of hand allograft recipients in different periods were examined by using flow cytometry.The recipients before transplantation were as the control groups.Results The first day after transplantation,the levels of CD3＋、CD3＋CD4＋、CD3＋CD8＋、CD3＋HLA-DR＋T cells and ratio of CD4/CD8 all began to descend.The 3th to 5th day after transplantation,the levels were the lowest.The 8th day,the levels of CD3＋、CD3＋CD4＋、CD3＋CD8＋、CD3＋HLA-DR＋T cells and ratio of CD4/CD8 were eventually rised and go to stable 15th day after transplantation.But the levels of CD3＋、CD3＋CD4＋and value of CD4/CD8 were still lower than those of the contols,and the levels of CD3＋CD8＋、CD3＋HLA-DR＋T cells were higher distinctly.Conclusion Dynamic changes of T cell subsets and active T cells in peripheral blood of composite tissue transplantation-hand allograft recipients were accordant with that of renal allograft recipients with stable period and with stable clinical state of the hand transplantation recipients.