4.A Comparative Study on Sensitivity of HE,IHC and RT-PCR in Detection of Breast Cancer Metastases in Axillary Lymph Nodes
Bo CHEN ; Yunfei LU ; Jian ZENG ; Sheng HE
Chinese Journal of Bases and Clinics in General Surgery 2003;0(03):-
Objective To compare the sensitivity of HE,immunohistochemistry (IHC) and RT-PCR in detection of breast cancer metastases in axillary lymph nodes.Methods Twenty female patients with newly diagnosed and clinically node-negative breast cancers underwent modified radical mastectomy, including a complete axillary lymph node dissection. The ages of the patients ranged from 31 years to 65 years, and the diagnosis of breast cancer was approved by pathological finding. Two hundred and thirty-nine axillary lymph nodes were found in these 20 patients. Metastases in axillary lymph nodes were explored by HE, cytokeratin 19 IHC and RT-PCR for cytokeratin 19 respectively. Results Seven(2.9%) lymph nodes were found to have metastatic cancers by HE in 3 patients,all nodes were found in level Ⅰ. Metastatic cancers were found in 13 (5.4%) nodes by IHC in 7 patients,11 nodes in level Ⅰ and 2 nodes in level Ⅱ; and 52(21.8%) nodes were found to contain tumor cells by RT-PCR in 14 patients,30 nodes in level Ⅰ and 22 nodes in level Ⅱ. All of 7 histologically(HE) positive nodes were found to contain tumor cells by IHC and RT-PCR. Among 232 histologically(HE) negative nodes,6(2.6%) nodes were found to contain tumor cells by IHC,and 45(19.4%) nodes were found to contain tumor cells by RT-PCR, all 6 IHC positive nodes showed the expected 460-base pair products on gel electrophoresis ( P
5.Expression of interleukin-17 and retinoid-related orphan nuclear receptor gamma t in cornea of experimental keratomycosis
Zong-sheng, ZENG ; Xiao-li, HAN ; Jian-zhang, HU
Chinese Journal of Experimental Ophthalmology 2013;(7):653-658
Background In the past few decades,the balance of Th1/Th2 is often used to explain the immune mechanisms of fungal infection and fungal disease.More recently,a novel subset of CD4+ effector Th cells has been found to participate in anti-fungal infection response.However,whether Th17 is involved in the immune response in fungal keratitis is unclear up to now.Objective Present study was to investigate the expression change of Th17 type cytokine and its specific transcription factor,retinoid-related orphan nuclear receptor gamma t (RORγt),in the cornea of Fusarium solani keratitis.Methods Ninety-six clean BALB/c mice were divided into Fusarium solani keratitis model group and control group by randomized digital table.Fusarium solani keratitis models were established by epikeratophakia-assisted corneal epithelial erasion and interlayerly injection of 5 μl (1 × 106 CFU/ml) Fusarium solani solution in the right eyes,and the equal volume of PBS was injected in the same way in the control group.10% KOH wet film was used to examine the fungal hyphea and funga strain was identified by inoculation.The corneas were examined under the slit lamp microscope 1 day,3,5,7 days after modeling and the inflammatory response was scored based on the criteria of Wu and Hu.The histopathological examination of corneas was performed in the time points above.Real time fluorescence quantitative PCR was used to detect the expression levels of interleukin-17 (IL-17) mRNA and RORγt mRNA in the corneas.The expression of IL-17 protein in the corneas was detected by ELISA.The use and raise of the mice followed the Statement of Association for Research in Vision and Ophthalmology.Results The inflammatory scores were 3.2±0.8,6.6± 1.1,9.4± 1.1 and 6.8±0.8 in 1 day,3,5,7 days after modeling,showing a significant difference among them (F =89.786,P =0.010).The inflammatory scores were higher in the third and seventh day than that in the first day (P<0.05),but they were significantly lower than that in the fifth day (P<0.05).The infiltration of inflammatory cells showed a coincident tendency with the score.The expressing levels of IL-17 mRNA (2-ΔΔCt) in the corneas were 4.12±0.73,20.72±1.81 and 14.16±1.88 in 3,5,7 days after modeling,with statistically significant differences in comparison with those in the control group (P<0.01),and the expression level was significantly higher in the fifth day than those in the first,third and seventh day in the model group(P<0.01).The expression levels of IL-17 protein (ng/g) were significantly increased 1 day,3,5,7 days in the model group compared with the control group (P<0.01).A similar change was found in the expression of RORγt mRNA to that of IL-17 mRNA.Conclusions Expressions of IL-17 and its transcription factor RORγt upregulate in the fungal keratitis and has an association with inflammatory degree,which suggests that Th17 subset may play an important role in the immune responses of fungal keratitis.
6.Effect of Low-Dose Hydrocortisone on Expression of Hippocampus Nuclear Factor-Kappa B,Inhibitory Kappa Bin Lipopolysaccharide-Induced Septic Rats
zhi-yuan, WU ; jian-sheng, ZENG ; xun-mei, FAN
Journal of Applied Clinical Pediatrics 1992;0(06):-
Objective To investigate the effect of low-dose hydrocortisone(HC)on hippocampus nuclear factor kappa B((NF-?B)),I?B expression in lipopolysaccharide(LPS)induced septic rats and the role of NF-?B signal transcription pathway in pathogenesis.Methods Fifty-four rats were randomly divided into 3 groups: control group(A group,n=6),model group(B group,n=24),low-dose HC treatment group(C group,n=24).The septic rat model was established by intraperitoneal injection LPS(1 mg/kg),as the intervention by caudal vein injection low-dose HC(6 mg/kg),each of B and C group was subdivied into 2,8,16,24 hours respectively after LPS injection(n=6).At serial time points,the animals in each group were sacrificed,brain tissue samples were harvested to determine NF-?B,I?B expression by immunhistochemistry in hippocampus.Results In B group: NF-?B expression was up regulated compared with A group(P
7.Significance of Change of Gastric Mucosal Partial Pressure of Carbon Dioxide in Rabbit Model with Septic Shock
jian-sheng, ZENG ; xun-mei, FAN ; su-yun, QIAN
Journal of Applied Clinical Pediatrics 1992;0(06):-
Objective To observe the change of gastric mucosal-arterial partial pressure of carbon dioxide gap [p(g-a)(CO2)] in septic shock rabbit.Methods Sixteen anesthetized and mechanically ventilated rabbit were randomly assigned to 2 groups:shock group(n=8) and control group(n=8).The rabbit in shock group were challenged with intravenous injection of 2 mg/kg Lipopolysaccharides from Escherichia coli.The rabbit in control group were intravenous injection of normal saline solution.Mean arterial pressure(MAP) and heart rate were continuously recorded by multichannel physiologic recorder.Cardiac index(CI) and superior mesenteric blood flow index(SMBFI) were continuously monitored by doppler flowmeter.Gastric mucosal partial pressure of carbon dioxide [pg(CO2)] was evaluated by gas tonometry every 10 min.Arterial and venous blood gas analysis,hemoglobin,and lactate levels were measured every 1 hour.Results The parameters remained stable in control group,but the parameters changed significantly in shock group.Compared with baseline levels,2 hours after Lipopolysaccharides infusion in shock group,MAP decreased from(78?5) mmHg to(50?2) mmHg(1 mmHg=0.133 kPa)(F=145.3 P
8.Meloxicam versus indomethacin in the prevention of heterotopic ossification after total hip arthroplasty
Yirong ZENG ; Linyang JIAN ; Wenjun FENG ; Jie LI ; Feilong LI ; Sheng HE
Chinese Journal of Tissue Engineering Research 2013;(39):6867-6874
BACKGROUND:In order to avoid heterotopic ossification after total hip arthroplasty, nonsteroidal anti-inflammatory drugs are commonly used for prevention.
OBJECTIVE:To compare the effect of meloxicam and indomethacin in the prevention of heterotopic ossification after total hip arthroplasty.
METHODS:Fifty-one patients who treated in the Department of Orthopedics, the First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine from 2010 to 2011 were col ected. Among the 51 patients, nine patients were treated with bilateral total hip arthroplasty, and al the patients had total hip arthroplasty with the posterior-lateral approach. The patients were divided into the control group and the experimental group according to the drugs used after replacement, and the patients in the two groups were administered with indomethacin sustained-release tablet 25 mg+omeprazole capsule 20 mg or meloxicam tablet 15 mg after replacement.
RESULTS AND CONCLUSION:There were no significant differences in the incidence of heterotopic ossification, pain, modified D’Aubigne and Postel scores after replacement between two groups (P>0.05). But, the gastrointestinal adverse reactions of the experimental group were less than those of the control group. The application of meloxicam only can effectively avoid the heterotopic ossification and release pain. Consequently, we recommend meloxicam as postoperative drug for the prevention of heterotopic ossification and pain remission fol owing total hip arthroplasty.
9.Proteomic analysis of gastric mucosa in chronic gastritis rats of Pi-Wei damp-heat syndrome treated by sanren decoction: an experimental study.
Sheng-Yin LIAO ; Jun ZENG ; Ai-Yao WANG ; Jian-Yong CHEN
Chinese Journal of Integrated Traditional and Western Medicine 2013;33(1):76-80
OBJECTIVETo study the expressions of gastric mucosal proteins in chronic gastritis (CG) rats of Pi-Wei damp-heat syndrome (PWDHS), to investigate the pathogenesis correlated to CG rats of PWDHS, to observe the differential expressions of gastric mucosal proteins in CG rats of PWDHS, and to investigate the mechanisms of Sanren Decoction (SD) for treating CG rats of PWDHS.
METHODSTotally 36 male SD rats were adaptable fed for 3 days and randomly divided into 3 groups, i.e., the normal control group, the CG of PWDHS rat model group (abbreviated as the model group), and the SD treatment group, 12 in each group. The CG of PWDHS rat model was prepared by composite factors. The gastric mucosal protein was separated using two-dimensional gel electrophoresis technique, and stained by Coomassie brilliant blue. The protein spots expressed differently were analyzed by PDquest 8.0 software. The protein spots expressed differently was identified by MALDI-TOF/TOF-MS.
RESULTSThe protein spots were 1 025 +/- 3 9, 994 +/- 51, 1 087 +/- 33 in the normal control group, the model group, and the SD treatment group respectively detected from two-dimensional gel electrophoresis profiles. Compared with the normal control group, there were 74 protein spots differentially expressed in the model group, 30 spots up-regulated and 44 spots down-regulated. Compared with the model group, there were 75 protein spots differentially expressed in the SD treatment group, 49 spots up-regulated and 26 spots down-regulated. Five protein spots differentially expressed were successfully identified, i.e., heat shock protein 72 (HSP72), heat shock protein 60 (HSP60), protein disulfide-isomerase (PDI), malate dehydrogenase (MDH), and unnamed protein.
CONCLUSIONSThe pathogenesis of CG of PWDHS may be correlated to energy metabolism disturbance and stress. The mechanisms of SD for treating it may possibly adjust differential expressions of gastric mucosal proteins.
Animals ; Drugs, Chinese Herbal ; therapeutic use ; Gastric Mucosa ; metabolism ; Gastritis ; diagnosis ; drug therapy ; metabolism ; Male ; Medicine, Chinese Traditional ; Phytotherapy ; Proteome ; metabolism ; Rats ; Rats, Sprague-Dawley
10.Effect of Estrogen on Expression of Osteoblast Apoptosis Related Genes Induced with Serum Hungry
Xiao-ming TANG ; Fu-xing PEI ; Sheng-fu LI ; Jiancheng ZENG ; Yaoming ZHANG ; Zhongqian LIU ; Jian PANG
Chinese Journal of Rehabilitation Theory and Practice 2006;12(11):960-962
ObjectiveTo explore the mechanism of estrogen inhibiting osteoblast apoptosis induced with serum hungry.MethodsOsteoblasts of the second or third generation from newly born SD rats calvaria were divided randomly into three groups: control group,serum hungry group,serum hungry with estrogen group.Cells of each group were incubated for 1,2,3,5,7 or 14 d,and then were stained immunohistochemically.The rates of positive cells of each group were analyzed.ResultsThere was a little positive expression of Bax,Bcl2 and Fas in control group.The expression of Bax and Fas were significantly increased(P<0.05)in serum hungry group,peak time was 14 d,but the expression of Bcl-2 were not affected.Compared with that of serum hungry group,the expression of Bax and Fas significantly decreased(P<0.05) in serum hungry and estrogen group,peak time was still 14 d,while that of Bcl-2 increased(P<0.05).ConclusionSerum hungry can increase the expression of Bax and Fas in osteoblast,that can be inhibited by estrogen.Estrogen can also increase the expression of Bcl-2 in osteoblast.All of these may play a role in inhibiting osteoblast apoptosis induced with serum hungry.