1.Expression of cyclooxygenase-2 and epidermal growth factor receptor in astrocytomas and its clinical significance
Hongtao ZHANG ; Miao TIAN ; Chunming XIU ; Jie LIU ; Guangqiang CUI ; Yunbo WANG ; Shumao PAN
Journal of Chinese Physician 2010;12(2):185-187
Objective To investigate the expression of cyclooxygenase-2 (COX-2) and epidermal growth factor receptor (EGFR) in astrecytomas, as well as the correlation between them. Methods The expression of COX-2, EGFR and PCNA were respectively detected by immunohistochmical (S-P) method in 68 astrocytomas and 5 cases normal brain tissue. Proliferation index (PI) was calculated and the correlation of COX-2, EGFR and PI was analyzed. Results COX-2 and EGFR were negative expression in normal brain tissue. The positive expression rate of COX-2 and EGFR in high grade astrocytomas was significantly higher than that in low grade astrocytomas(73.53% vs 44. 18% ,67.65% vs 38.24%, P <0. 01 ), and the PI was significantly higher than that in low grade astrocytumas as well as normal brain tissue(46.11 ± 10. 68vs 23. 04±6. 25,4. 52±0. 95, P <0. 01 ). The PI in COX-2 positive group was higher than that in negative group( P <0. 01 ). The positive expression rate of COX-2 in the group with EGFR positive expression was higher than that in the negative group. Conclusions The expression of COX-2 and EGFR was related to pathological feature of astrocytomas. COX-2 may promote the proliferation of tumor cells. There was a static correlation between the expression of EGFR and COX-2 in astrocytomas. EGFR signal transduction probably modulated the expression of COX-2 in astrocytomas cells.
2.The clinical study of nuclear factor-kappa B expression and counts of lymph vessels in laryngeal sqnamous cell carcinoma and polyps of vocal cord tissues
Shan LIANG ; Zhuori LI ; Zhulin YANG
Journal of Chinese Physician 2010;12(2):182-184
Objective To study the expression of nuclear factor-kappaB (NF-κB) and the counts of lymph vessels in laryngeal squarnous cell carcinoma and polyps of vocal cord tissues, and explore their clinicopathologic significance and correlation in the course of laryngeal carcinoma. Methods SP immuno-histochemical method was used to detect the expression of NF-κB and the counts of lymph vessels on the routinely paraffln-embedded sections of the specimens from 50 cases laryngeal squamous cell carcinoma and 10 cases of polyps of vocal cord tissues. Results The positive rate of NF-κB and the counts of lymph ves-sels in laryngeal carcinoma[60. 0% ,( 13.3±3.4)/HP]were significantly higher ( P <0. 05 and P <0. 01respectively) than those in polyps of vocal cord tissues[10.0 % ,(6. 1±3. 8)/HP]. The positive rate of NF-κB and the counts of lymph vessels in well differentiated adenocarcinoma and cases without metastasis were significantly lower( P < 0. 05, P <0. 01 ), compared with poor-differentiated adenoearcinoma and ca-ses with metastasis. The counts of lymph vessels in the NF-κB positive cases were significantly higher than thoseinNF-κBnegativecases[(14.9±4.1)/HPvs (9.8±3.1)/ HP, P <0.01] . Conclusions The expression of NF-κB and the counts of lymph vessels might be important markers to be used to monitor the progression, biological behaviors, metastatic status and prognosis of laryngeal carcinoma. NF-κB might pro-mote lympoangiogenesis in laryngeal squnmous cell carcinoma tissues.
3.Effects of acute lymphoblastic leukemia children bone marrow mesenchymal stem cells on the drug tolerance of K562 cell strains
Zhaoxia WANG ; Yuxin ZHAO ; Yawei ZOU ; Chongjun DONG ; Chunguang XIAO
Journal of Chinese Physician 2010;12(6):775-778
Objective To study the effect of acute lymphoblastic leukemia (ALL) children bone marrow mesenchymal stem cells (MSCs) on the resistance of K562cell atd mechanism in vitro.Method MSCs were obtained from AL children bone marrow after derivation, cultivation and identification.The coculture of MSCs and K562 and K562 suspension were established.Effects of MSCs on the growth of K562 cells were investigated in vivo.The two kinds of cells treated with different concentration of adriamycin (ADM) and the rate of apoptosis was evaluated by flow cytometry.Cell cycle was determined by flow cytometry.RT-PCR was used to detect Bcl-2 and Bax in K562 cells.Result Compared with the cell growth curve of K562 alone, the K562 cell co-cultured with MSCs grew slower and the exponential phase of growth was not obvious.The apoptosis index of the K562 cells co- clutured with MSCs was (9.19 ±0.53)% examined by flow cytometry, and that of the K562 cells alone was 4.00 ± 0.37% respectively( P < 0.05 ).The percentage of cells at G0/G1 phase was (50.2 ± 2.26) % and that at S phase was (37.03 ± 3.50) % in the group of K562 alone, but those of the K562 cells co - cultured with MSCs were (80.95 ± 3.83) % and ( 17.40 ± 1.50)% respectively( P <0.05).The result of RT-PCR suggested expression of Bcl-2/Bax of the K562 cell co-cultured with MSCs was higher than K562 alone.Conclusion ALL children MSCs suppressed the growth of K562 cell in vitro.Adhesion made K562 depress sensitive to ADM.The mechanism was perhaps caused by adhesion with MSCs, K562 cell cycle was changed and related to Bcl-2 gene high level expression.
4.Analyze the relationship between serum XCL1 levels and the immune injury in CHB patients
Journal of Chinese Physician 2010;12(6):768-771
Objective To explore the relevance between serum XCL1 levels and liver damage in hepatitis B patients.Methods The serum concentration of XCL1 was detected by enzyme linked immu-nosorbent assay (ELISA).Peripheral blood T-cell subsets were detected by flow cytometry (FCM).Liver function was assayed by automatic biochemistry analyzer, hepatitis B antigen/antibody semi-quantitative index was detected by time-resolved fluorescence analyzer, and HBV-DNA load was detected by automatic fluorescence quantitative PCR.Results Serum concentration of XCL1 in control group ( n = 20), mild chronic hepatitis B (CHB) group ( n =29), moderate CHB group ( n =20) and severe CHB group ( n =26)were (8.24±1.94) pg/ml, (10.99±1.94) pg/ml, (12.83 ±2.59) pg/ml, (13.72 ±3.13) pg/ml,respectively.The concentration of XCL1 in all CHB groups was significantly higher than control group ( P< 0.05 ).The concentration of XCL1 in severe CHB group was significantly higher than mild CHB group (P < 0.05).XCL1 was positively correlated with ALT, AST, TBIL and DBIL, and the coefficients were (r =0.463、 0.472、 0.413、 0.440, P <0.01 ), respectively.The serum XCL1 levels in hepatitis B virus with low load group was lower than hepatitis B virus with high load group.The percentage of CD4 + T in hepatitis B virus with low load group and high load group were (41.26 ± 11.33)%, (33.01 ± 5.96)%,and the difference was statistically significant.Conclusion Serum concentrations of XCL1 were closely related to the degree of liver inflammation in hepatitis B patients.XCL1 may be involved in the process of chronic hepatitis B.
5.Comparison of two techniques for endothacheal intubation in mice
Younian XU ; Pu MA ; Zhao ZHANG ; Shihai ZHANG
Journal of Chinese Physician 2010;12(6):740-742
Objective To explore a safe and efficient method for endotracheal intubation in mice.Methods 60 BALB/c mice were random divided into 2 groups, direct intubation under direct vision group, ( n = 30) and direct intubation under light by transillumination group ( n = 30).After successful tracheal intubation at the first judgment, mouse received a tracheal instillation of 3 mg/kg lipopolysaccharide.Bronchoalveolar lavage (BAL) was then performed three times with 0.8 ml sterile saline.Exudate cells were centrifuged and stained with Wright's fluid.Whether the intubation indeed succeeded was judged by the amount of neutrophil infiltrated into the lungs.The mortality after intubation, the time consumed for each successful tracheal intubation and the times of attempting for intubation in two groups were assessed to approve the efficiency of two techniques.Results Eight vs.two mice died within 24 hours in two groups respectively.The time consumed for each successful tracheal intubation and success rate for the first time for two groups were (92.6 ±23.4)s vs (64.0±20.1)s and 53.3% vs 86.7% respectively, which the consumed time was significantly different, while the final success rate almost the same for the two groups.Conclusion Both direct intubation under direct vision and intubation under light by transillumination can successfully intubate the tube into the trachea.But direct intubation under direct vision by transillumination is more efficient and safe in endotracheal intubation in mice.
6.Study on the changes and clinical significance of the DNT cells and T lymphocyte subtype in children with infectious mononucleosis
Huiqin XUE ; Lei ZHU ; Rui ZHAO ; Huimin GUO ; Weili DU
Journal of Chinese Physician 2010;12(6):772-774
Objective To study on the changes of the DNT cells and T lymphocyte subtype in children with infectious mononucleosis (IM) and its clinical significance.Methods The flow cytometry (FCM) was used to detected the DNT cells and other T lymphocyte subtype in 48 cases of IM.Results The study showed that DNT cells( 9.39 ± 4.89 )% were greatly increased in comparison with normal controls (NC) (4.26 ± 1.68)% ( P <0.01 ).CD4 cells(21.45 ±9.87)% were decreased ( P <0.01 ) and CD8 cells increased in comparison with NC (32.43 ± 5.07) % ( P < 0.01 ).Conclusion DNT cells and T lymphocyte subtype can be used to evaluate the immune function of children with infectious mononucleosis (IM) and provide guidance for adoptive immunotherapy.
7.Detection of plasma PAF-AH and G994T single nucleotide polymorphism research in type 1 diabetes
Yuanyuan ZHANG ; Guanqi GAO ; Changgui LI
Journal of Chinese Physician 2010;12(6):752-755
Objective To investigate the plasma PAF-AH level in initialed type 1 diabetes patients and the association between the PAF-AH gene polymorphism and type 1 diabetes mellitus. Methods155 initialed type 1 diabetes patients and 138 controls were selected in this study. Plasma PAF-AH was determined by PAF-AH Assay kit, and the PAE-AH gene G994t genotypes were detected by PCR. ResultsThe activity of plasma PAF-AH in the patients with type 1 diabetes was significantly lower than that in the controls[ (20.64 ± 6.23)nmol/(min · ml) vs (28.56 ± 4. 11)nmol/(min · ml), P <0.05). Compared with control group, the frequencies of the GT genotype on G994T polymorphism between type 1 diabetes patients and controls were not significantly different(8.4% vs 7. 2%, P >0. 05). ConclusionThe activity of plasma PAF-AH in the patients with type 1 diabetes was significantly lower than controls. The G994T polymorphism of PAF-AH gene was not related to type 1 diabetes mellitus.
8.Effects of Ang-( 1-7 ) on the apoptosis of cultured endothelial cells induced by Ang Ⅱ
Huiyu YANG ; Zhiming YANG ; Yunfei BIAN ; Maolian LI ; Nana ZHANG ; Fen GAO ; Chuanshi XIAO
Journal of Chinese Physician 2010;12(6):748-751
Objective To investigate the effect of Ang-(1-7) on the apoptosis in human umbilical vein endothelial cells (HUVECs) induced by Ang Ⅱ.Methods HUVECs were isolated and cultured.Cultured HUVECs were incubated for 24 h with Ang-(1-7), Ang Ⅱ, Ang-(1-7) A-779, Ang-(1-7) + Ang Ⅱ, A-779 + Ang Ⅱ + Ang-( 1-7), respectively.Cultured HUVECs without incubating stimulator were chosen as controls.The apoptosis of endothelial cells were detected by flow cytometry.Results The apoptosis of endothelial cells in HUVECs were upregulated by AngⅡ ( 10-6 mol/L) (25.60% ±3.17% vs 2.32% ±0.24%, P <0.005).Compared with the AngⅡ group, Ang-(1-7) dose-dependently inhibited the apoptosis of endothelial cells in HUVECs ( (20.04% ± 2.21% ,16.04% ± 1.32 %, 10.04% ±2.05,7.79% ±1.50% vs AngⅡ group 25.60% ±3.17%, P <0.05 , P <0.05).The effects of Ang-(1-7) could be blocked by A-779 (23.37% ±0.75% vs 20.04% ± 2.21%, 16.04% ± 1.32,10.04% ± 2.05% ,7.79%± 1.50%, P < 0.05 ).Conclusion Ang-(1-7) can attenuate the apoptosis of endothelial cells induced by Ang Ⅱ in HUVECs in a dose-dependent manner.The effects of Ang-(1-7) could be blocked by A-779( P<0.05).
9.Rosiglitazone and ATRA on gastric cancer SGC7901 cell line proliferation in vitro
Kaihong XIE ; Guoliang LI ; Zhongshan ZHANG ; Xiaochun ZENG
Journal of Chinese Physician 2010;12(6):743-747
Objective To investigate the influence of PPARγ excitomotor RSG and ATRA on gastric cancer SGC7901 cell line proliferation in vitro and its potential mechanism study.Methods Human gastric cancer SGC7901 cell line was cultured in vitro.Experiment samples were divided to blank group,10μmol/L ATRA group, 12.5μmol/L RSG group, 25μmol/L RSG group, 10μmol/L ATRA + 25μmol/L RSG group.Proliferation inhibitory effect was determined by MTI assay.Flow cytometry was used to detect cell cycle, H.E stain was used to observed micrography alteration.Expression of PPARγ protein in gastric cancer cells were measured by immunohistochemistry.PPARγ mRNA in gastric cancer cells were measured by RT-PCR.Results ATAR at concentration 10μmol/L, RSG at 12.5 μmol/L and RSG at 25 μmol/L could inhibit the proliferation of SGC7901 cells in a dose-and time-dependent, and when both agents were combined for 72h, growth inhibition ratio was (29.73 ± 0.69) %.Flow cytometry analysis revealed a cell cycle arrest at G1 and S phase, and when both agents combined, S% was (12.87 ± 0.35 )%, cell micrography tended to be normal when both agents combined.Up-regulation of PPARγ protein and PPARγ mRNA expressions were also observed, those effects were enhanced when both agents combined, and grey scale ratio was 0.646.Conclusion The ATRA and RSG could significantly induced growth inhibition of human gastric cancer SGC7901 cell, which may be associated with cell cycle arrest and inducing differentiation, activation of PPARγ protein and PPARγ mRNA expression.Synergistic effect could be caused by the combined use of the two agents.
10.The immunohistochemical analysis of AGEs, SP-A in the lung of diabetic rats
Wei ZHAO ; Tingwei ZHANG ; Ruilin WANG ; Baosheng ZHANG ; Hong ZHANG
Journal of Chinese Physician 2010;12(6):732-735
Objective To approach the changes of advanced glycosylated end products (AGEs),surfactant proteins A (SP-A) in the lung of experimental diabetic rats and their relationship. Methods 48 male SD rats were divided into diabetes mellitus (DM) group and control group, each group with 24 rats.The DM rat model was made by injecting streptozocin (60mg/kg) into caudal vein. The rats were killed and the lung was individually taken out at the end of 4, 12 and 20 weeks after the models were established. The changes of AGEs, SP-A in rats lung were observed with immunohistochemical assay and the images were analyzed( black is minimum of gray, white is maximum of gray ). Results We observed a great quantity of AGEs positive cells in the alveolar epithelial cells, bronchial mucosal epithelium, angio-endothelial cell and smooth muscle cells of the DM rats. The average gray (AG) was inferior to that of the controls(4weeks 93.92 ± 7.92 vs 104. 75 ± 8. 20; 12 weeks 76. 25 ± 6. 76 vs 93.50 ± 7.56; 20 weeks 47.63 ± 7.96 vs 142. 38 ± 19. 76; P <0. 05) and decreased with the DM course. In the 4 weeks DM rats, there were a few SP-A positive cells in the type Ⅱ alveolar epithelial cells, Clara cells and alveolar macrophage cells. In the 12 and 20 weeks DM rats, there were a great many CTGF and TGF-β1 positive cells. The AG was inferior to that of the controls( 12 weeks 75.63 ± 6. 70 vs 110. 50 ± 13.20;20 weeks 47.38 ± 4. 84 vs 97. 25 ± 9. 87; P < 0. 01 ). Conclusion With the progress of diabetes, DM rats' pulmonary alveolar type Ⅱ cells injury appeared, that might be related with the deposition of AGEs.