1.Fluorescence in situ hybridization and immunohistochemistry for detecting HER-2 gene amplification and protein expression in breast cancer tissue
Haifeng ZHOU ; Xueliang WU ; Xibin SUN ; Likun WANG ; Wanping LIANG
Chongqing Medicine 2014;(11):1299-1301
Objective To analyze and compare the fluorescence in situ hybridization(FISH) and immunohistochemical(IHC) for detecting HER-2 gene amplification and protein expression in breast cancer tissues .Methods 110 cases of breast cancer from Janu-ary 2008 to May 2012 receiving the modified radical mastectomy were selected .The resected breast cancer tissue was detected by FISH and IHC and the detected results were performed the comparative analysis .Results Among 110 cases of breast cancer tissue , 25 cases(22 .73% ) were the HER-2 protein expression(+ + + ) ,44 cases(40 .00% ) were(+ + ) ,26 cases(23 .64% ) were(+ ) and 15 cases(13 .64% ) were(-) .Among 110 cases ,the gene amplification was in 28 cases(25 .45% ) and no gene amplification was in 82 cases(74 .55% ) .The positive(+ + + ) of the IHC detection was coincident with that of FISH ,and the negative(+ /-) of the IHC detection was also coincident with that of FISH ,there was statistical difference between the suspicious positive of the IHC de-tection and the results of FISH (P<0 .05) .But the total coincidence of the IHC detection results and FISH test results was 89 .29%(25/28) ,and the two detection methods had the positive correlation (χ2 =84 .89 ,P<0 .01) .Conclusion The positive and negative expression of the IHC detection has better consistency with that of the FISH detection .However ,the coincidence of the IHC suspi-cious positive expression and the FISH results is poor ,indicating that the suspicious positive sample of the IHC detection needs to be detected by the FISH detection .
2.Relationship between expression of Her-2 gene and the transfer number of axillary lymph node and its influence on prognosis
Haifeng ZHOU ; Yuhong FAN ; Xueliang WU ; Likun WANG ; Xibin SUN ; Wanping LIANG ; Yi ZHOU
Chinese Journal of Postgraduates of Medicine 2014;37(26):3-5
Objective To investigate the relationship between expression of Her-2 gene and the transfer number of axillary lymph node and its influence on prognosis.Methods A total of 351 cases with primary breast cancer from January 2008 to January 2011 were selected.The expression of Her-2 gene was detected by immunohistochemical method,and analyzed the relationship between it and the transfer number of axillary lymph node and its influence on prognosis.Results The expression of Her-2-,+,++ had no correlation with the transfer number of axillary lymph node (x2 =3.757,1.650,2.379,P > 0.05),while the expression of Her-2 +++ had correlation with the transfer number of axillary lymph node (x2 =8.681,P < 0.05).The 2 years survival rates in the expression of Her-2-,+,++,+++ were 77.01% (201/261),85.00% (34/40),29.17%(7/24),1 1.54% (3/26),and which in the expression of Her-2--+ was significantly higher than that in the expression of Her-2 ++-+++ (x2 =61.605,P < 0.01).The transfer number of axillary lymph node was 0 and 1-3,the 2 years survival rate in the expression of Her-2--+ was significantly higher than that in the expression of Her-2 ++-+++,and there was significant difference (x2 =61.605,14.747,P < 0.05).The transfer number of axillary lymph node was 4-9 and ≥ 10,there was no significant difference in the 2 years survival rate between the expression of Her-2--+ and Her-2 ++-+++ (x2 =3.691,3.482,P > 0.05).Conclusions The expression of Her-2-,+,++ has no correlation with axillary lymph node metastasis,and the 2 years survival rate in the expression of Her-2--+ is higher than that in the expression of Her-2 ++-+++,while Her-2-has no difference with Her-2 + in prognosis.While the transfer number of axillary lymph node ≤ 3,the expression of Her-2 gene may be an important prognostic factor.
3.MicroRNA-143 inhibits proliferation and migration as well as invasion in esophageal cancer cell line ECA109
Wenxue WU ; Wanping LIU ; Hebo LIU ; Jie CHEN ; Yanyan SUN ; Chuantao ZOU ; Yanmei LI
Journal of International Oncology 2016;43(11):801-805
Objective To investigate the functions of microRNA-1 43 (miR-1 43)in esophageal cancer cell line ECA1 09.Methods ECA1 09 cells were transfected with negative control (NC),miR-1 43 mimics or miR-1 43 inhibitors.3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT)assay was per-formed to evaluate the growth of ECA1 09 cells after transfection.Annexin V-FITC /PI apoptosis test kit was used to detect early apoptosis rate in ECA1 09 cells.Transwell migration and invasion assays were conducted to compare the migration and invasion capacity of ECA1 09 among different groups.Real-time PCR and Western blotting were used to analyze the mRNA and protein alteration after transfection.Results Three and four days after transfection,compared with NC (absorbance value:0.90 ±0.02 and 1 .09 ±0.07),miR-1 43 mimics inhibited ECA1 09 cell proliferation (absorbance value:0.66 ±0.05 and 0.80 ±0.04),while miR-1 43 inhibi-tors promoted cell proliferation (absorbance value:1 .1 3 ±0.09 and 1 .51 ±0.08),with statistical signifi-cances (F =49.1 6,P =0.000;F =1 00.34,P =0.000).Early-stage apoptosis rates of ECA1 09 transfected with NC,miR-1 43 mimics and miR-1 43 inhibitors were 3.42% ±0.72%,1 1 .63% ±1 .1 5% and 0.94% ± 0.1 0%,respectively,with statistical significance (F =1 51 .61 ,P =0.000).Meanwhile,compared with NC (migration cell number:336 ±1 3,invasion cell number:1 47 ±1 6),miR-1 43 mimics inhibited cell migration (1 48 ±1 6)and invasion (75 ±1 0),while miR-1 43 inhibitors promoted cell migration (51 0 ±1 4)and inva-sion (238 ±1 6),with statistical significances (F =470.99,P =0.000;F =90.04,P =0.000).Compared with NC (1 .00 ±0.00),miR-1 43 mimics down-regulated mRNA (relative expression level 0.22 ±0.08)and protein expression (relative expression level 0.46 ±0.08)of K-ras,whereas miR-1 43 inhibitors up-regulated mRNA (1 .55 ±0.1 2)and protein expression (1 .33 ±0.05)of K-ras (F =1 31 .36,P =0.000;F =88.1 7, P =0.000).Conclusion miR-1 43 functions as a tumor suppressor in esophageal cancer cell line ECA1 09, probably by down-regulating K-ras expression.
4.An association analysis on rs1412125 at HMGB1 with atrial fibrillation
Li LI ; Zhen DU ; Jun YANG ; Jiawang DING ; Jin SUN ; Jiong ZHANG ; Wanping CHEN ; Song LI ; Zhiyang LV ; Yong CHEN
The Journal of Practical Medicine 2015;(6):884-887
Objective To investigate the association of rs1412125 at high-mobility group box 1 (HMGB1) gene with atrial fibrillation (AF) in Chinese Han population. Methods 396 patients with AF and 726 control subjects were recruited to this study. A case-control association analysis was applied to assess the as-sociation between rs1412125 in HMGB1 and AF. Results There were no significant differences in rs1412125 and the frequency of genotype distribution between the study group and the control group (allelic association:object P value was 1.05E-06, adjust P value was 0.176; genotypic association: additive P value was 0.146, dominant P value was 0.162, and recessive P value was 0.998). The interactions analysis showed that rs1412125 allele C increased the risk of AF in male subjects with rheumatic heart disease (adjust P value was 2.07E-04, rectify OR = 8.20, and 95%CI: 2.70-25.0). Conclusions There is no independent genetic association between rs1412125 at HMGB1 and atrial fibrillation. However , the interactions between rs1412125 and both gender and rheumatic heart disease might increase the risk of atrial fibrillation.
5.The effects of Shenfu decoction on expression of cold-inducible RNA-binding protein in rats with traumatic brain injury treated by mild hypothermia
Guan WANG ; Ying CAI ; Hongsheng SUN ; Wanping HAO ; Dongting YANG ; Taoli WANG ; Pengfei BIE ; Jiayu LIU ; Xueyan WANG
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2016;23(5):479-483
ObjectiveFrom the changes of expression of cold-inducible RNA-binding protein (CIRP) in rats with traumatic brain injury under mild hypothermia treatment with Shenfu decoction as a subsidiary, to speculate the mechanism of protective effect of the decoction on the injury.Methods Ninety Sprague-Dawley (SD) rats were divided into three groups by random number table: non-transfection control group, adenovirus mediated immune flourescent reverse transcription virus group (blank AD5-GFP transfection group) and adenovirus mediated immune flourescent reverse transcription virus carrying CIRP silent expression gene group (AD5-GFP-CIRP-SiRNA transfection group), 30 rats in each groups. Then, each group was subdivided into three subgroups: model group, traditional Chinese medicine (TCM) low and high dose groups, 10 rats in each subgroup. After the mild hypothermia treatment for 48 hours, in the TCM low dose group and high dose group, a dose of TCM 1 mL/kg and 5 mL/kg was injected via a tail vein into the rat respectively, while in the model group, 1 mL/kg normal saline was injected into the same vein, once a day for consecutive 2 days in all the groups. Before modeling in the blank AD5-GFP transfection group and AD5-GFP-CIRP-SiRNA transfection group, virus transfection models were reproduced at first by one-time intrathecal injection of 0.1 mL AD5-GFP and 0.1 mL (1×1010 pfu/mL) AD5-GFP-CIRP-SiRNA virus vector respectively, and in model group, 0.1 mL normal saline was given. The rat cortex, hippocampus and hypothalamus part were collected, the brain cell apoptosis was detected by transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL), the CIRP mRNA expression in the cortex, hippocampus and hypothalamus part was measured by reverse transcription-polymerase chain reaction (RT-PCR), the protein expressions of rat sarcoma protein Raf, Ras, extracellular signal-regulated kinase (ERK), phosphorylation ERK (p-ERK), mitogen activated protein kinase (MEK), p-MEK were determined by Western Blot.Results The brain tissue cell apoptosis indexes (AI) in the cortex, hippocampus and hypothalamus part in TCM low, high dose group of non-transfection control and blank AD5-GFP transfection group were lower than those in model group, and the expressions of CIRP mRNA were higher than those in model group, there were no significant differences in AI and CIRP mRNA in the cortex, hippocampus and hypothalamus between model, TCM low and high dose groups of AD5-GFP-CIRP-SiRNA transfection group, but AI was significantly higher and CIRP mRNA was significantly lower than that in corresponding subgroups of AD5-GFP transfection control group and blank AD5-GFP transfection group. Western Blot detection showed that: Raf/Ras, p-MEK/MEK protein expressions revealed no statistical significant differences in different parts of each group (allP > 0.05), the p-ERK/ERK protein expression in the cortex, hippocampus, and hypothalamus part was significantly lower in TCM low and high dose group than that in the model group of non-transfection control group and blank AD5-GFP transfection group, the degree of descent in the TCM high dose group being more significant (the cortex: non-transfection control group was 7.2±1.0 vs. 15.3±1.8, AD5-GFP transfection group was 8.1±0.7 vs. 16.2±1.5; hippocampus part: non-transfection control group was 6.6±0.8 vs. 14.7±2.0, AD5-GFP transfection group was 6.8±1.0 vs. 14.9±1.3; hypothalamus part: non-transfection control group was 9.4±1.1 vs. 12.7±1.7, AD5-GFP transfection group was 10.6±1.3 vs. 9.4±1.1, allP < 0.05). There were no significant statistical differences in p-ERK/ERK protein expression in above brain parts between AD5-GFP-CIRP-SiRNA transfection subgroups (allP > 0.05).Conclusions The Shenfu decoction used in rats with brain trauma under treatment of mild hypothermia is possibly by promoting CIRP over-expression, lowering ERK expression and inhibiting the initiation of signal transduction of the secondary transcription factor phosphorylation, thereby the neural cell apoptosis is decreased and play a subsidiary role of anti-apoptosis of mild hypothermia.
6.Herbal textual research on Bunge pricklyash seed
Yutong DUAN ; Junhan YANG ; Ming RAN ; Jiaxin SUI ; Lingjie MENG ; Wanping SUN
China Pharmacist 2024;27(2):357-364
By consulting the relevant literature and historical herbal literature,the name,origin and harvesting,efficacy and indications,as well as nature,taste,and meridians of Bunge pricklyash seed were reviewed."Jiaomu"as a medicinal herb name was first recorded in the"Compendium of Materia Medica Annotations",and later generations also used"Jiaomu"as a proper name.Bunge pricklyash seeds are the seeds of the Rutaceae plant Sichuan Bunge pricklyash seed or green Bunge pricklyash seed.They are harvested when they mature in autumn from August to October,and can be processed by net,stir-frying or salt.It can promote diuresis and reduce swelling,treat kidney deficiency and tinnitus,and is mainly used to treat bloating in the abdomen.It is non-toxic or slightly toxic.This article traced the origin of Bunge pricklyash seed,studied the history of medicine,clarified the original traditional Chinese medicine properties of Bunge pricklyash seed,and provided scientific basis for the correct clinical use and the national formulation of quality standards for this herb.
7.Varieties and Prescription Characteristics of Chinese Patent Medicines for Stroke in China
Jingdan ZHANG ; Wanping SUN ; Xiaoxia LIN ; Shuo ZHANG ; Xue ZHANG ; Jiahui YAO ; Yiming LIU ; Ming XIE
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):270-274
ObjectiveTo explore the listed varieties and prescription characteristics of Chinese patent medicines for stroke in China, explore the medication rules of Chinese medicine for stroke, and provide guidance for further clinical research and development of Chinese patent medicines. MethodsExcel 2021 and the Ancient and Modern Medical Record Cloud Platform (V2.3.5) were used to systematically mine and analyze the varieties and prescriptions of Chinese patent medicines for stroke in China. ResultsA total of 244 Chinese patent medicines (two for different dosage forms of the same prescription), 1 736 approval documents for Chinese patent medicines, 792 manufacturers, and 83 varieties of protected Chinese patent medicines were finally included in the database. The top three dosage forms were capsules (75), pills (53), and tablets (42). There were 28 Chinese patent medicines for stroke in the National Essential Drug Catalogue (2018), 129 in the National Essential Medical Insurance, Industrial Injury Insurance and Maternity Insurance Drug Catalogue (2023), and 4 in the National Non-prescription Drug Catalogue. Among the 138 prescriptions screened out, Chinese patent medicines mainly treated stroke patients with the syndrome of Qi deficiency and blood stasis. The top three most frequent medicinal herbs were Chuanxiong Rhizoma (63), Pheretima (47), and Salviae Miltiorrhizae Radix et Rhizoma (47). The medicinal herbs used were mainly warm, pungent, with the meridian tropism to the liver meridian. The correlation analysis showed that the herb pair with the highest support was Astragali Radix-Chuanxiong Rhizoma, and that with the highest confidence was Carthami Flos-Chuanxiong Rhizoma. Five herb combinations were identified based on the cluster analysis. ConclusionThe Chinese patent medicines for stroke mainly treat patients with the syndrome of Qi deficiency and blood stasis. The medicinal herbs used in the prescriptions mainly have the functions of activating blood and resolving stasis, extinguishing wind and stopping convulsions. Drug compatibility usually focuses on activating blood and resolving stasis, as well as expelling phlegm and opening orifices. This review of the varieties and prescription characteristics of Chinese patent medicines for stroke helps optimize clinical decision-making, guide drug research and development, promote medical research and scientific progress, and provide more effective support and guarantee for the treatment of stroke patients.