1.Experimental study on the effects of tumor necosis factor on the cultured human umbilical vein endothelial cells
Yingbo YANG ; Zhengguo WANG ; Peifang ZHU
Journal of Third Military Medical University 1988;0(06):-
The effects of tumor necrosis factor (TNF) on the activity of plasmogen acitva-tor inhibitor (PAD and the morphology of the cells were observed in the original passage of cultured human umbilical vein endothelial cells (HUVEC). It was found that the TNF effect on the PAI was determined by the dosage of TNF, that is, 500U/ml of TNF could significantly activate PAT activity while 2500U/ml markedly inhibit it. Morphdogical observation canfirmed that 2500U/ml of TNF could injure HUVEC to form a kind of cells whose morphology was quite similar to that of circulating endothelial cells originating from peripheral blood.These findings suggest that TNF can stimulate HUVEC to increase the PAI acivity but too large a dosage of TNF could injure the metabolic function and structure of HUVEC.
2.PREPARATION AND CHARACTERIZATION OF MONOCLONAL ANTIBODY AGAINST SYNTHETIC PEPTIDES OF HUMAN TOLL-LIKE RECEPTOR 4
Qingwu YANG ; Peifang ZHU ; Zhenggu WANG
Medical Journal of Chinese People's Liberation Army 1982;0(01):-
An immunogenic peptide sequence of B cell dominant epitope of human Toll like receptor 4 (TLR4) was selected basing on predication of B cell epitope of TLR4.bALB/c mice were immunized with synthetic peptide of TLR 4 and one clone (B4) of hybridoma cells was picked out by ELISA and limited dilution. High titer McAb prepasations against TLR 4 were obtained from ascites fluid of BALB/c mice infravenously inoculated with the McAb inducing hybridoma cells.The titer of ascites fluid was 1∶ 100 000 .The subclass of the McAb was IgG2,k.The specificity of McAb was identified. by indirect ELISA,and a single strong blot was shown in molecular weights of 90 Kd.using Western blot analysis.
3.A study on the dose-effects relationship of underwater blast injury
Zhihuan YANG ; Peifang ZHU ; Jianxin JIANG
Medical Journal of Chinese People's Liberation Army 1983;0(02):-
Objective To study the dose-effects relationship of underwater blast injury. Methods Sixty-one adult mongrel dogs were exposed to underwater explosion of 200g, 500g, and 1 000g of TNT, respectively. The physical parameters of blast wave were recorded by PCB pressure transducers (USA). Survival or death was observed 6 hours after injury. Correspondingly, the relationship between the physical parameters and injury severity was analyzed. Results Twenty-three among 61 dogs died after injury, the mortality rate was 37.70% (23/61). The main reasons of death were severe lung bleeding, lung edema, perforation of gastrointestinal tract, and rupture of liver and spleen. Half lethal (5/10) impulse of underwater blast wave was 337.7?14.3kPa?ms. The impulses resulting in mild, moderate, severe, and critical underwater blast injuries were 140.46?34.2, 199.2?25.2, 247.8?69.6, and 478.7?183.8kPa?ms, respectively. Conclusion The physical parameters were well related to the injury severity in underwater blast. The early main treatment consist of active and effective management of severe lung injuries, perforation of gastrointestinal tract, and rupture of liver and spleen.
4.Correlation of the expressions of scavenger receptor and CD14 in murine alveolar macrophages with lung injury
Ce YANG ; Jianxin JIANG ; Peifang ZHU
Chinese Journal of Tissue Engineering Research 2005;9(6):222-224
BACKGROUND: There exists a close relationship between the expression of scavenger receptor(SR) and CD14 and the endotoxic lung injury. To further detect the expression of these two receptors and their relationship is helpful to disclose the receptor mechanism of lung inflammatory disequilibrium so as to provide a possible theoretical foundation for sepsis therapy.OBJECTIVE: To investigate the expression of CD14 and SR in murine alveolar macrophages and their effect after trauma and traumatic endotoxin challenge.DESIGN: Randomized and controlled experimental study.SETTING: Fourth Laboratory(a state key laboratory), Trauma, Burns and Combined Injury, the Research Institute of Field Surgery, Daping Hospital,the Third Military Medical University of Chinese PLA.MATERIALS: The experiment was completed in the Research Institute of Field Surgery of Daping Hospital of the Third Military Medical University of Chinese PLA from August 2001 to November 2001. Totally 63 Kunming mice with murine alveolar macrophages were used in the experiment.METHODS: The experimental animals were randomly divided into three groups including control group, trauma group and traumatic endotoxin challenge group. The model mice with both femur fracture and endotoxin challenge were used. Endotoxin at a dose of 5 mg/kg was injected one hour after fracture via tail vein and the dynamic expression of CD14 and SR in murine alveolar macrophages (AM) was detected.RESULTS: Six hours after trauma, the expression of CD14 and that of SR in AM showed the tendency of up-regulation and down-regulation respectively. Furthermore, the post-traumatic endotoxin challenge induced CD14up-regulation and SR down-regulation drastically. Correlation analysis indicated that CD14 and SR showed a significantly negative relationship.CONCLUSION: The study suggested that the dual modulation of CD14 and SR in AM is possibly related with the change of "self-control" to "out of control" of inflammatory reactions in the lungs after endotoxic challenge. The dual regulation of the effective antagonists CD14 and SR are helpful in further understanding lung injury and its repairing mechanism.
5.Changing of pulmonary surfactants after Severe Steam Inhalation Injury in Rabbits
Zhiyuan LIU ; Ngao LI ; Peifang CHU ; Chongcheng YANG ; Jintang SHUI
Journal of Third Military Medical University 1984;0(01):-
Surface tension(in terms of hysteresis), lecithin content, PaO2 and lung water content of both bronchial irrigation fluid and lung homogenates were determined after severe inhalation injury in rabbits. In accompany with the decrease of PaO2 and increase of lung water content, there was a remarkable dropping of pulmonary surfactant content as illustrated by progressive increase of surface tension and decrease of lecithin content in both bronchial irrigation fluid and lung homogenate. It was proposed that dropping of pulmonary surfactant played an important role in the pathogenesis of pulmonary edema after inhalation injury. Therefore, successive determinations of lecithin content or and surface tension of bronchial irrigation fluid might serve as an early diagnosis as well as a pro-gnostis aid in severe inhalation injuries.
6.Unbalance and involvement of Th17/Tr cells in patients with cervical cancer
Ting FU ; Peifang YANG ; Zhijun JIAO ; Jianxin ZHANG
Chinese Journal of Microbiology and Immunology 2011;31(4):312-315
Objective To detect the levels of Th17 and regulatory T(Tr)cells in Imripheral blood mononuclear and tumor tissue from patients with cervical cancer and analyze the relationship hetween Th17 and Tr cells in cervical cancer progression.In addition.the significance of the Th17/Tr cells ratio in cervical cancer pathogenesis was discussed.Methods The expression levels of Th17 and Tr cells were determined by flow cytometry from 32 patients with cervical cancer and 15 health people.The mechanism of involvement of Th17 and Tr cells proportionality in cervical cancer pathogenesis and the correlation between Th17 and Trwas assessed by bivariate correlation analysis.Results The expression levels of both Th17 and Tr in patients were higher than control groups,especially in late stage patients Th17 and Tr proportionality lower than early group,and there was a negative correlation between them.Conclusion Th17 and Tr cells proportionality may be involve in the development of cervical cancer so as to provide novel strategies for tumor immunotherapy.
7.Primary malignant lymphoma of the breast: imaging characteristics and correlation with histopathologic findings
Peifang LIU ; Lu YIN ; Yun NIU ; Jianmei YANG ; Runxian BAO
Chinese Journal of Radiology 2001;0(01):-
Objective To evaluate the mammographic characteristics of primary lymphoma of the breast and to correlate the imaging features with the histopathologic findings.Methods We retrospectively reviewed records in 27 patients with histologically proven primary malignant lymphoma of the breast.Mmammography was performed before surgery in 14 cases.The clinical, pathologic, and imaging features were retrospectively reviewed and correlated in these 14 patients.All patients were female, with the mean age of 41.5 years (range 28-56).The lesion was in the right breast in 7 patients, the left in 6, and bilateral in 1.Results Histology revealed 13 Non-Hodgkin lymphoma (NHL) and 1 Hodgkin lymphoma (HL).In primary lymphoma, mammography revealed a unilateral solitary mass in 9 cases, unilateral multiple masses in 2 cases, bilateral multiple masses in 1 case, and unilateral diffusely increased opacity with skin thickening in 2 cases.The diameter of the masses ranged from 0.7 cm to 5.0 cm (mean 2.6 cm). Of the total 18 masses, the margins of the masses were well defined in 13, and partially defined in 5.Neither calcifications nor spiculations nor adjacent skin retraction were seen.Of the 13 NHL, histologic examination showed diffuse NHL in 12 cases and follicular NHL in 1.No mammographic features were identified that could helped distinguish follicular from diffuse NHL.Conclusion Primary lymphoma of the breast usually presents with clinical and mammographic findings mimicking a carcinoma or benign tumor.Although the clinical and imaging characteristics may suggest the possibility of breast NHL, none of the findings are pathognomonic.The final diagnosis depends on histopathology.
8.Quantitative study of cochlear hair cells injuries in guinea pigs exposed to blast underpressure
Chaojun LI ; Zhaohua LIU ; Peifang ZHU ; Zhengguo WANG ; Cheng YANG ; Haibin CHEN ; Jihong ZHOU ; Xin NING
Chinese Archives of Otolaryngology-Head and Neck Surgery 2006;0(03):-
OBJECTIVE To explore the characteristics of cochlear hair cell injuries in guinea pigs exposed to blast underpressure(BUP). METHODS The guinea pigs were killed 14 days after exposure to experimental BUP. Their basilar membranes were stained by silver nitrate and the hair cell injuries were quantitatively assessed using the light microscope. RESULTS The outer hair cells(OHCs) of the guinea pigs clearly appeared to be injured following exposure to the experimental BUP at a peak underpressure varying between -22.4kPa and -63.3kPa. The most obvious injury was in the second turn and OHC loss was smallest in the first row and more severe in the second and third rows. OHC loss was seen in all of three experimental groups of animals following exposure to BUP. Furthermore, the higher the peak of underpressure, the more obvious the injury of OHCs. Quantitative morphological analysis of cochlear hair cells showed that the total OHC loss rates in all the experimental groups of guinea pigs exposed to BUP were significantly higher than that of control group animals(P
9.Value of breast imaging reporting and data system in Chinese breast cancer screening
Yanhong AN ; Zhaoxiang YE ; Yi LI ; Chun QING ; Jianmei YANG ; Yumei ZHAO ; Peifang LIU ; Kexin CHEN
Chinese Journal of Radiology 2011;45(4):353-357
Objective To study the value of breast imaging reporting and data system (BI-RADS)in Chinese breast cancer screening. Methods A total number of 3483 women participated in breast cancer screening with mammography in Hexi district in Tianjin from August to December 2009, which was organized by ministry of public health. BI-RADS assessment categories and recommendations were compared with histological findings. The precision, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were calculated. Results Among 3483 screening mammography cases, 267 were almost entirely fat breast, 1245 were scauered fibroglandular, 1890 were dense and 81 extremely dense.There were 1011 patients(29.0%) with category 1, 1741 (50.0%) with category 2, 383 (11.0%) with category 3, 59 patients(1. 7%) with category 4 and 16 (0. 5%) with category 5 according to BI-RADS assessment categories. Totally, 71 women with 77 lesions were confirmed by histological examinations. There were 29 malignant and 48 benign lesions. The diagnostic precision, sensitivity, specificity of BI-RADS were 63. 6% (49/77) , 93. 1% (27/29) and 45.8% (22/48) . The general PPV of BI-RADS was 50. 9%(27/53). The PPV of categories 0, 4, 5 were 25.0% (1/4), 36. 4% (12/33) and 87. 5% (14/16). The NPV of categories 2 and3 were90.9% (10/11), 100.0% (12/12). Conclusions B1-RADS is of much value in assessing the breast malignancy. It is applicable in Chinese breast cancer screening.
10.Primary expression and identification of TLR4 extracellular fragment in Pichia pastoris
Qingwu YANG ; Ling MOU ; Peifang ZHU ; Zhengguo WANG ; Jianxin JIANG ; Jingzho WANG
Journal of Third Military Medical University 1984;0(01):-
Objective To express and identify of Toll-like receptor 4 (TLR4) extracellular fragment in Pichia pastoris and to provide basis for the related researches. Methods The human TLR4 extracellular cDNA fragment was amplified by PCR and after confirming by DNA sequence analysis, was inserted into the Pichia pastoris expression vector pPIC9K containing AOX1 promoter and lead sequence of ? factor gene to form a pPIC9K/TLR4 extracellular recombinant expression plasmid. The recombinant plasmid was transformed into GS115 and high copies transformants were screened with G418. After being identified by colony PCR, transformants were cultured in flasks and induced by 1% methanol. The expression products were analyzed by SDS-PAGE and were identified by Western blot analysis. Results The sequencing showed that TLR4 extracellular gene was identical to that in Genbank. The analysis for the recombinant plasmid DNA digested by enzyme demonstrated that the recombinant expression plasmids of pPIC9K/TLR4 extracellular were constructed successfully. After screening by G418, 200 high-copy-number of transformants were acquired. The special TLR4 extracellular gene segment was obtained by identification with colony PCR, which showed that TLR4 extracellular gene of 5 clones were inserted into Pichia pastoris. After methanol induction for 4 d, the expression proteins came up to 50.35% of total proteins in medium supernatant as shown by SDS-PAGE. Western blot analysis proved that the expression protein could bind to TLR4 monoclonal antibody specifically. Conclusion TLR4 extracellular protein is expressed and identified successfully by Pichia pastoris system, which can provide basis for the researches of further screening of high expression colony, protein purification, and functional research.