2.Triple staining of immunohistochemistry.
You-zhi YU ; Min LIN ; Wei-cheng XUE ; Qiu-jing SONG ; Dan-hua SHEN
Chinese Journal of Pathology 2005;34(4):244-245
3.Effect of Shenfu injection on brain apoptosis and expression of HO-1 during focal cerebral ischemia reperfusion injury in rats
Lei BAO ; Hua SHEN ; Zheng ZHANG ; Ying XU ; Mingzhou MA ; Hui CHENG ; Xi SONG ; Haidong QIN
Chinese Journal of Emergency Medicine 2012;21(9):992-996
Objective To investigate the effects of Shenfu injection ( SF,a Chinese herbal medicine preparation made of Codonopsis pilosula and Aconitum carmichaeli) on the cell apoptosis of focal cerebral ischemic-reperfusion injured rats and the expression of heme oxygenase-1 (HO-1). Methods Forty-two male Sprague-Dawley rats used for producing unilateral brain ischemia reperfusion model were randomly divided into three groups:sham operation group ( Sham group),ischemia reperfusion group ( IR group),and SF Injection group (SF group).The model of focal cerebral ischemia-reperfusion injury was induced by transient occlusion of middle cerebral artery (ischemia for 2 h,and reperfusion for 3,6 h respectively).In SF group,SF ( 10 mg/kg) was intraperitoneally injected duri(n)g reperfusion.Cell apoptosis rate in brain tissue was detected by the technique of Annexin-V-PI double staining and was counted in flow cytometer.Expression of HO-1 in brain was measured by RT-PCR,while the pathological and ultra structure changes of cerebral tissue were also observed.Results Cell apoptosis rate of brain tissue were significantly higher in IR group than that in Sham group (P <0.01 ),while SF group had less significant changes in cell apoptosis rate, HO-1 level of brain tissue than IR group (P < O.01 ).The ultra structure change of brain tissue was less in SF group than that in IR group.Conclusions During early stage of brain IR injury,SF inhibits cellular apoptosis and in turn protects the brain from injury which is attributed to the increase in HO-1 expression induced by SF.
4.The difference of T lymphocyte subsets between mild type and severe type in patients infected with the 2009 pandemic influenza A (H1N1) virus and its clinical significance
Jun WANG ; Jun JIN ; Fang HUANG ; Jianan HUANG ; Cheng JI ; Yueping SHEN ; Hua XU
Chinese Journal of Emergency Medicine 2011;20(2):193-197
Objective To study the changes of subgroups of peripheral blood T lymphocytes in the patients infected with the 2009 pandemic influenza A ( H1N1 ) virus of different severity type. Method A total of 66 patients infected by H1N1 evidenced by RT-PCR admitted from September 2009 to January 2010 were divided into three groups: mild type ( B group, n = 47 ), cured patients of severe and critical severe type ( C group, n = 14) and died patients ( D group, n =5), according to the severity and prognosis. A total of 20 healthy volunteers served as control group( A group). Peripheral blood lymphocyte count, CD3+,CD4+ and CD8+ T lymphocyte count were detected by flow cytometry at the different time points. Fever duration and H1N1 virus negative time were compared. Statistical analysis were performed by using SAS version 9.13 software and the data were processed with ANOVA and SNK test. Results Lymphocyte count, CD3+,CD4+ and CD8+ T lymphocyte count declined in the early period in all the groups, and there were significant differences compared with A group (P<0. 05), while rised with the clinical progression in group B and C,and those of C group were lower than B group ( P < 0.05 ), but those of D group were always low. Fever duration and H1N1 virus negative time were (4.4 ± 1.6) days vs. (4.4 ± 1. 4) days, ( 12.9 ± 3. 1 ) days vs.( 10.2 ± 2.6) days and ( 15.2 ± 7.3 ) days vs. ( 13.3 ± 2.9 ) days respectively, and there were significant differences among the three groups ( P < 0.05 ). Conclusions The cellular immune function was seriously damaged when patients were infected with H1N1. Further more, the changes of lymphocyte count, CD3+ , CD4+and CD8+ T lymphocyte count were tightly related with the degree of severity and prognosis. These findings can be used for clinical diagnosis and treatment.
5.Clinicopathologic analysis and expression of cyclin D1 and p53 of ovarian borderline tumors and carcinomas
Hui-Lin SHAO ; Dan-Hua SHEN ; Wei-Cheng XUE ; Yi LI ; You-Zhi YU ;
Chinese Journal of Obstetrics and Gynecology 2001;0(04):-
Objective To study the clinicopathologieal features and expression of cyclin D1 and p53 in epithelial ovarian tumors,and to investigate the correlation between pathogenesis of ovarian cancer and epithelial borderline tumors.Methods Fifty four cases of ovarian borderline tumors and 45 cases of ovarian carcinomas from the People's Hospital,Peking University were reviewed retrospectively.The clinical data and pathological findings were analyzed.Immunohistochemical study of cyclin D1 and p53 was performed in all 99 cases.Results(1)In borderline tumors,the age of patients ranged from 14-82 (mean age=42.5)years.International Federation of Gynecology and Obstetrics(FIGO)stage of borderline tumors was stage Ⅰ in 48 cases,stage Ⅱ in 3 cases,and stage Ⅲ in 3 cases.In ovarian carcinomas,the age of patients ranged from 26-80(mean age=53.5)years.FIGO stage of carcinoma was stage Ⅰ in 6 cases, stage Ⅱ in 8 cases,stage Ⅲ in 26 cases,and stage Ⅳ in 5 cases.In follow-up of 54 cases with borderline tumors the 5-year survival rate was 98% and of 45 cases with carcinomas a 5-year survival rate of 51% was noted.(2)In 54 cases of borderline tumors,mucinous types accounted for 56%(30/54)and serous types accounted for 30%(16/54).There were 5 cases with micropapillary pattern,3 cases with peritoneal implants,3 cases with lymph node involvement,6 cases with microinvasion,one case with intraepithelial carcinoma,and one case with mural nodules.In 45 cases of carcinomas,serous carcinoma was the most (49%,22/45).The remainder included 3 cases of mucinous types,8 cases of endometrioid types,6 cases of transitional cell types,3 cases of mixed phenotype and 3 cases of undifferentiated types.(3) Overexpression of cyclin D1 and p53 was observed in 31%(14/45)and 56%(25/45)of ovarian carcinomas, respectively.There was a significant association between p53 overexpression and tumor grade.In the borderline tumor group,69%(37/54)had overexpression of cyelin D1 and 6%(3/54)had overexpression of p53.There were significant differences in expression of cyclin D1 and p53 between conventional serous borderline tumors and high-grade serous carcinomas(cyclin D1:91% vs 26%;p53:0 vs 58%).However, micropapillary serous borderline tumors and low-grade serous carcinomas showed remarkably similar expression of cyelin D1 and p53.Conclusions Epithelial ovarian borderline tumors are distinct from ovarian cancer in clinical progress and prognosis,and histological types.Overexpression of cyclin D1 is common in ovarian borderline tumors and low grade carcinomas.And overexpression of p53 is more common in high grade ovarian carcinomas.Conventional serous borderline tumors are distinct from high-grade serous carcinomas in pathogenesis.Micropapillary serous borderline ovarian tumors may be closely related to low grade serous carcinomas.
6.Study on extraction process of polysaccharide from Sargassum fusiforme by enzymatic treatment.
Hua-fang ZHANG ; Jing-shun JIN ; Rong-mei TAN ; Cheng SHEN
China Journal of Chinese Materia Medica 2006;31(22):1860-1862
OBJECTIVETo investigate the optimal extraction process of polysaccharides from S. fusiforme by enzymatic treatment.
METHODThe optimum extraction conditions were obtained by the experiment with the orthogonal design. The content of polysaccharides of S. fusiforme was determined by spectraphotometry.
RESULTThe amount of enzyme and temperature significantly affected total polysaccharides of S. fusiforme.
CONCLUSIONThe optimum extraction conditions include the addition of 1. 2 x 10 (4) U x 100 g(-1) enzyme into water at pH 4. 5, and the subsequent treatment for 10 min while the temperature is maintained at 45 degrees C.
Cellulase ; metabolism ; Hydrogen-Ion Concentration ; Polysaccharides ; analysis ; isolation & purification ; metabolism ; Sargassum ; chemistry ; Technology, Pharmaceutical ; methods ; Temperature
7.Effects of naloxone on pneumocyte apoptosis during pulmonary ischemia reperfusion injury
Zheng ZHANG ; Hua SHEN ; Ying XU ; Mingzhou MA ; Haibin NI ; Hui CHENG ; Xi SONG ; Lei BAO ; Haidong QING
Chinese Journal of Emergency Medicine 2010;19(5):507-510
Objective To investigate the effects of naloxone (Na) on pneumocyte apoptosis and expression of heme oxygenase-1 (HO-1) during ischemia reperfusion injury of lung in rats. Method Forty-two male Sprague-Dawley rats were made models of ischemia reperfusion injury of unilateral lung, and were randomly( random number) divided into three groups: sham operation group (Sh group), ischemia reperfusion group (IR group) and naloxone group (Na group). The hilus of lung was clamped for 45 minutes and the clamp was taken off to build the I/R model. After 3-6 hours reperfusion, naloxone in dose of 1 mg/kg was injected intra-peritoneally in rats of Na group. The rate of cell apoptosis in lung tissue was detected with the way of Annexin-V-PI in flow cy-tometer. The wet to dry weight ratio (W/D) of lung tissue was measured. The expression of HO-1 in lung was measured by using RT-PCR and the ultra-structure change of lung tissue was observed under electron microscope. Results The rate of pneumocyte apoptosis and W/D ratio of lung tissue were significantly higher in IR group than in Sh group (P < 0.01), and the rate of pneumocyte apoptosis and W/D ratio of lung tissue were negatively correlated with the expression of HO-1 mRNA in lung tissue. Compared with IR group, the rate of cell apoptosis and W/D ratio were lower and the expression of HO-1 mRNA was higher in Na group (P < 0.01). The ultra- structure changes of lung tissue were lessened in Na group than in IR group. Conclusions During early period of lung IR injury, HO-1 induced by naloxone can inhibit the cellular apoptosis and protect the lung tissue.
8.Study on the B cell linear epitopes of rabies virus CVS-11 nucleoprotein.
Xin-Jun LV ; Xin-Xin SHEN ; Peng-Cheng YU ; Hao LI ; Li-Hua WANG ; Qing TANG ; Guo-Dong LIANG
Chinese Journal of Virology 2014;30(3):253-256
To study the B cell linear epitopes of rabies virus CVS-11 nucleoprotein, peptides were synthesized according to the amino acid sequences of B cell linear epitopes. Linear epitopes predicted by bioinformatics analysis were evaluated with immunological techniques. Indirect enzyme-linked immunosorbent assay showed that titers of antibodies to peptides (355-369 and 385-400 residues of rabies virus CVS-11 nucleoprotein) were above 1:12 800 in mouse sera. The antibodies recognized denatured rabies virus CVS-11 nucleoprotein in Western blot analysis. Purified anti-peptide antibodies recognized natural rabies virus CVS-11 nucleoprotein in BHK-21 cells in indirect fluorescent antibody test. The 355-369 and 385-400 residues of rabies virus CVS-11 nucleoprotein were validated as B cell linear epitopes.
Amino Acid Sequence
;
Animals
;
Antibodies, Viral
;
immunology
;
Epitope Mapping
;
Epitopes, B-Lymphocyte
;
chemistry
;
genetics
;
immunology
;
Female
;
Humans
;
Male
;
Mice
;
Mice, Inbred BALB C
;
Molecular Sequence Data
;
Nucleoproteins
;
chemistry
;
genetics
;
immunology
;
Rabies
;
immunology
;
virology
;
Rabies virus
;
chemistry
;
genetics
;
immunology
9.Effects of surface roughness of bone cements on histological characteristics of induced membranes.
Hai-Xiao LIU ; Hua-Zi XU ; Yu ZHANG ; Gang HU ; Yue SHEN ; Xiao-Jie CHENG ; Lei PENG
China Journal of Orthopaedics and Traumatology 2012;25(8):662-666
OBJECTIVETo explore surface roughness of bone cement and surround tissue on histological characteristic of induced membranes.
METHODSBone cements with smooth and rough surface were implanted in radius bone defect, intramuscular and subcutaneous sites of rabbits, and formed induced membranes. Membranes were obtained and stained (HE) 6 weeks later. Images of membrane tissue were obtained and analyzed with an automated image analysis system. Five histological parameters of membranes were measured with thickness,area,cell density,ECM density and microvessel density. Double factor variance analysis was used to evaluate the effect of the two factors on histological characteristics of induced membranes.
RESULTSMembranes can be induced by each kind of bone cement and at all the three tissue sites. In histological parameters of thickness,area and micro vessel,there were significant differences among the membranes induced at different tissue sites (P = 0.000, P = 0.000, P = 0.000); whereas, there were no significant differences in histological parameters of cell density and ECM density (P = 0.734, P = 0.638). In all five histological parameters of membranes, there were no significant differences between the membranes induced by bone cements with different surface roughness (P = 0.506, P = 0.185, P = 0.883, P = 0.093, P = 0.918).
CONCLUSIONSurround tissue rather than surface roughness of bone cements can affect the histological characteristics of induced membranes. The fibrocystic number, vascularity, mechanical tension and micro motion of the surround tissue may be closely correlated with the histological characteristics of induced membranes.
Animals ; Bone Cements ; Female ; Membranes ; cytology ; Rabbits ; Radius ; cytology ; Surface Properties ; Tissue Engineering ; methods ; Tissue Scaffolds