1.The relation between Child-Turcotte classification and pathology, diagnosis, prognosis and fibrosis index
Journal of Third Military Medical University 2003;0(10):-
Objective To study the relation between Child Turcotte classification and pathology, diagonsis, prognosis and fibrosis index as well as its significance. Methods The levels of hyaluronic acid(HA), type Ⅲ procollagen(PCⅢ), Laminin(LN) and type Ⅳ collagen(Ⅳ C) were detected by enzyme linked immunoadsordent assay(ELISA) or radioimmunoassay(RIA). Pathomorphology was observed in 68 patients with cirrhosis. Results Level of HA in serum was positively correlated with cirrhotic severity, but other fibrosis indexes could not reflect cirrhotic severity. Child Turcotte classification was concordant with the pathological changes. The concordance rate of Child Turcotte classification B or C patients with pathologic diagnosis was up to 97.8%. The recovery rate of Child Turcotte classification A patients was up to 95.5%. Fatality rate of Child Turcotte classification C patients was up to 96.9%. Conclusion Child Turcotte classification is closely correlated with the severity of hepatic fibrosis, severity of pathological changes in liver and prognosis of patients. It is of clinical value in the reflection of severity of hepatic cirrhosis.
2.Proportion and difference of neural stem cells and neurons from different embryonic mouse brain tissues
Fenglan ZHANG ; Lujun YANG ; Zhicheng XIAO
Chinese Journal of Tissue Engineering Research 2017;21(21):3364-3369
BACKGROUND:At present, mouse embryonic neural stem cells (NSCs) culture has been skillfully operated by many labs, but there are differences existing about which part are dissociated to get NSCs. Embryonic 14 days (E14) mouse brain tissues are widely used for culturing NSCs, but there are less studies about the detailed percentage and difference of NSCs separated from different brain tissues. OBJECTIVE: To test the proportion and difference of NSCs and neurons percentage from E14 mouse whole brain, cortex and forebrain, providing quantized data for optimizing the isolation of high-purity NSCs. METHODS:E14 C57BL/6 mouse whole brain, cortex and forebrain tissues were separated and dissociated into single cells that were adherently cultured for 3.0-4.0 hours and labeled by DAPI. Then the cells were immunostained with NSCs specific marker, Nestin, and neuron specific marker, Tuj1, to identify NSCs and neurons percentage by calculating Nestin+/DAPI and Tuj1+/DAPI. In addition, real-time PCR assay was used to test Nestin and Tuj1 mRNA expression in the E14 mouse whole brain, cortex and forebrain. RESULTS AND CONCLUSION: (1) Immunocytochemical results showed that there were a large amount of Nestin+ and Tuj1+ cells in the whole brain, cortex and forebrain of E14 mice. NSCs percentage in the forebrain was obviously higher than that in the whole brain (P < 0.01) and cortex (P < 0.05), while the percentage of neurons in the forebrain was significantly lower than that in the whole brain (P < 0.05) and in the cortex (P < 0.001). (2) Real-time PCR results showed that the Nestin mRNA expression in the forebrain was significantly higher than that in the whole brain (P < 0.05) and slightly higher than that in the cortex (P > 0.05); the Tuj1 mRNA expression in the forebrain was significantly lower than that in the whole brain (P < 0.05) and in the cortex (P < 0.05). These findings indicated that the forebrain had the most NSCs and the least neurons compared with the whole brain and the cortex. In summary, E14 mouse forebrain has the highest percentage of NSCs compared with the whole brain and cortex, which is a better source to obtain NSCs for the following cell culture experiments.
3.The clinical study of escharectomy in shock phase in coping with burn complications
Yaowen SUN ; Zhenxin ZHANG ; Lujun YANG
Journal of Clinical Surgery 1999;0(05):-
Objective To study the availability of escharectomy in shock phase and its role in preventing complications.Methods To make an analysis between the escharectomies and skin grafting of 79 cases in or beyond shock phase in the incidence of sepsis,visceral complications,MODS,mortality,healing time and the expenditure.Result The cases of the early operations revealed much better consequences than those performed beyond shock phase.Conclusion The escharectomy in shock phase proves to be available and significant in reducing post burn complications.
4.Construction and functional analysis of a bispecific antibody that targets TNF-α and ED-B.
Lujun LI ; Yanqun YANG ; Xueping HU ; Mian XIE ; Mengyuan LIU
Acta Pharmaceutica Sinica 2014;49(12):1665-73
In order to enhance the specificity of TNF-α monoclonal antibody to inflamed site, a bispecific antibody BsDb that targets TNF-α and the extra-domain B (ED-B) of fibronectin (FN) was constructed by covalently linking the anti-TNF-α single chain Fv antibody (TNF-scFv) and the anti-ED-B scFv L19 via a flexible peptide linker deriving from human serum albumin (HSA). ED-B is an antigen specifically expressed at the inflamed site. BsDb is expressed in E. coli, identified by immunoblot, and purified with affinity chromatography. This was followed by further examination of its bioactivities and pharmacokinetics. We demonstrated that BsDb retained the immunoreactivity of its original antibodies as it could simultaneously bind to TNF-α and ED-B and neutralize the biological action of TNF-α. In the collagen-induced arthritis mice model, BsDb selectively accumulate in the inflamed joint with a maximal uptake of (12.2 ± 1.50)% ID/g in a single inflamed paw and retain in the inflamed paw for at least 72 h. In contrast, BsDb showed a short serum half-life of (0.50 ± 0.05) h and a rapid clearance from normal tissues. The findings reported herein indicate that BsDb has good specificity to the inflamed site and low toxicity to normal tissues. BsDb is therefore likely to have greater clinical applications in the treatment of rheumatoid arthritis and other autoimmune diseases. This laid a stable basis for its preclinical study.
5.Incidental nodal irradiation with involved-field conformal radiotherapy for patients with thoracic esophageal squamous cell carcinoma
Kai JI ; Lujun ZHAO ; Chengwen YANG ; Zhenxing FENG ; Ping WANG
Chinese Journal of Radiation Oncology 2013;(1):30-34
Objective To quantify the incidental irradiation dose (ⅡD) to lymph node stations of esophagus when treating patients with T1-4N0 M0 thoracic esophageal squamous cell carcinoma (ESCC) with a dose of 60 Gy/30f.Methods Twenty-nine patients with medically inoperable T1-4N0M0 thoracic ESCC were treated with three-dimensional radiotherapy on involved-field.The conformal CTV was re-created using a 3 cm margin in the proximal and distal direction (following the course of the esophagus) beyond the barium esophagogram,endoscopic examination and CT defined GTV and a 0.5 cm margin in the lateral and anteroposterior directions of the CT defined GTV.The PTV encompassed 1 cm proximal and distal margins,0.5 cm radiaI margin on the basis of CTV.Cervical,mediastinal and abdominal lymph nodes were delineated respectively.Equivalent uniform dose (EUD) and other dosimetric paraneters were calculated for each nodal station.Nodal region whose metastasis rate is greater than 5% was considered a high risk lymph node subgroups.Results Under a 60 Gy dose prescription,the median Dmean and EUD,V40 and V50 were ≥40 Gy,≥85% and ≥75% in most of the high risk nodal regions.For the subgroups whose EUD were less than 40 Gy,most of the ⅡD of these regions was significantly associated with the length and location of esophageal tumor (r =0.892,P =0.000).Conclusions Lymph node stations nearby of ESCC received considerable ⅡD with involved-field irradiation which could control subclinical lesions.But more clinical studies should be needed.
6.Study on the acute toxicity of Tripterygium Hypoglaucum (Levl) Hutch solution in mice
Lujun YANG ; Houyong YU ; Ping HUANG ; Jian YUAN ; Jia CAO ;
Journal of Third Military Medical University 2003;0(17):-
Objective To explore the acute toxicity and LD 50 of Tripterygium Hypoglaucum (Levl) Hutch(THH) solution to provide information for safe clinical application. Methods After oral administration of THH solution in mice, the mortality and the physiological and pathological changes were observed. Results The LD 50 (95% confidence limit) of THH in male and female mice was 79 g/kg(69~89 g/kg) and 100 g/kg (90~112 g/kg), respectively. No marked pathological change of the organs was found. Conclusion According to the standard of grading of acute toxicity, THH solution belongs to the moderate class. Therefore, it is safe in clinical practice and has a wide application.
7.Role of [Ca~(2+)]i in the apoptosis of Jurkat T lymphoma cells induced by Tripterygium Hypoglaucum (Levl) Hutch alkaloids
Lujun YANG ; Wernera GRUNDLER ; Michael NUESSE ; Lin AO ; Jia CAO ;
Journal of Third Military Medical University 2003;0(17):-
Objective To explore the roles of [Ca 2+ ]i in the apoptosis of Jurkat cells induced by Tripterygium Hypoglaucum (Levl) Hutch alkaloids. Methods After Jurkat cells were stained with Indo 1 AM and PI, changes of [Ca 2+ ]i were assessed by flow cytometry at the single cell level. Results THH alkaloids could induce no changes of [Ca 2+ ] inside cytoplasm. Conclusion THH alkaloids induce the apoptosis of Jurkat cells via other pathways rather than via the [Ca 2+ ] pathway.
8.Tripterygium Hypoglaucum (Levl) Hutch alkaloids induce DNA strand breaks and DNA fragmentation in Jurkat T lymphoma cells
Lujun YANG ; Michael NUESSE ; Lin AO ; Jia CAO ;
Journal of Third Military Medical University 2003;0(17):-
Objective To investigate the effects of Tripterygium Hypoglaucum (Levl) Hutch alkaloids on the nuclear DNA strand breaks and DNA fragmantation in Jurkat T lymphoma cells to understand the mechanisms of the apoptosis. Methods After Jurkat cells were induced by the alkaloids, DNA stand breaks were labeled by TUNEL assay and DNA fragmentation was analyzed by DNA content analysis. Flow cytometry was performed to determine these phenomena. Results THH alkaloids could effectively induce DNA stand breaks and DNA fragmentation. Conclusion There are great changes in nuclear DNA in the apoptosis of Jurkat T lymphoma cells induced by THH alkaloids.
9.Effects of Tripterygium Hypoglaucum (Levl) Hutch alkaloids on cell cycle and time-dependent apoptosis of Jurkat T lymphoma cells
Lujun YANG ; Lin AO ; Michael NUESSE ; Ying XU ; Jia CAO ;
Journal of Third Military Medical University 2003;0(17):-
Objective To study the effects of the cytotoxicity, cell cycle and time dependent apoptosis of Jurkat T lymphoma cells induced by Tripterygium Hypoglaucum (Levl) Hutch (THH) alkaloids so ad to explore the mechanisms of the apoptosis induced by the THH alkaloids. Methods Cell vitality and cell proliferation were measured by Typan blue staining. Cell cycle and the time dependent apoptosis were determined by DNA staining, TUNEL labeling and flow cytometry. Results THH alkaloids could effectively inhibit cell proliferation of Jurkat cells and induce the G 1 arrest and could induce apoptosis in G 2/S phase first and then G 1 phase. Conclusion THH alkaloids can inhibit DNA synthesis, cell proliferation and can also induce apoptosis of Jurkat T lymphoma cells in all phases.
10.Study on X-ray induced frequency of micronucleated erythrocytes in mice bone marrow by flow cytometer
Bingbing HU ; Lujun YANG ; Shengxue LIU ; Jia CAO
Journal of Third Military Medical University 2003;0(22):-
Objective To detect the frequency of micronucleated polychromatic erythrocytes(fMNPCE) in mice bone marrow after irradiated with X-ray by flow cytometer(FCM) to study whether fMNPCE detecting method could become a rapid biodosimeter for early radiation damage. Methods BALB/c mice were randomly assigned to receive the X-ray irradiation at dose of 0.5,3.0,6.0 Gy(n=4 for each irradiation dose),and the bone marrow was collected in 6,12,24 h after irradiation.Another 4 mice were intraperitoneally injected with cyclophosphamide twice as positive control,of which the bone marrow was collected in 42 h after intoxication began and 4 treated with spurious radiation as normal control.Results fMNPCE in mice bone marrow increased remarkably in 24 h after exposure to 0.5 Gy X-ray(P