摘要
Objective:To detect the expression differences of Wnt signaling pathway related molecules β-catenin, Cyclin D1 and Dickkopf-1 (DKK1) in breast disease tissues, and to explore their application value in pathologically aided diagnosis of breast cancer.Methods:From January 2008 to August 2019, 90 cases of breast tissue specimens in the Cancer Center of Guangzhou Medical University were collected, including 30 cases of breast hyperplasia, 30 cases of breast intraductal carcinoma and 30 cases of breast invasive ductal carcinoma. The expressions of β-catenin, Cyclin D1 and DKK1 in breast tissue of each group were detected by immunohistochemistry. Oncomine database and KM plotter database were used to analyze the expression differences of β-catenin, Cyclin D1 and DKK1 in breast cancer and normal breast tissues and their relationships with survival prognosis of patients with breast cancer, and to verify the results of immunohistochemistry. Receiver operating characteristic (ROC) curve was used to evaluate the efficacies of each molecule in pathologically aided diagnosis.Results:There were statistically significant differences in β-catenin, Cyclin D1 and DKK1 expressions among breast hyperplasia, breast intraductal carcinoma and breast invasive ductal carcinoma ( χ2=7.766, P=0.021; χ2=24.133, P<0.001; χ2=11.585, P=0.003). The expression of β-catenin in breast invasive ductal carcinoma group was significantly higher than that in breast intraductal carcinoma group and breast hyperplasia group ( Z=-2.367, P=0.018; Z=-2.462, P=0.014). The expression of Cyclin D1 in breast invasive ductal carcinoma group and breast intraductal carcinoma group was significantly higher than that in breast hyperplasia group ( Z=-4.166, P<0.001; Z=-4.174, P<0.001). The expression of DKK1 in breast invasive ductal carcinoma group and breast intraductal carcinoma group was significantly higher than that in breast hyperplasia group ( Z=-3.090, P=0.002; Z=-2.923, P=0.003). The results of bioinformatics analysis showed that compared with normal breast tissue, the expression of β-catenin mRNA in invasive breast cancer tissue increased by 2.33 times ( t=15.242, P<0.001), the expression of Cyclin D1 mRNA in breast intraductal carcinoma tissue increased by 6.64 times ( t=7.152, P=0.006), while the expression of DKK1 mRNA in normal breat tissue was 3.41 times higher than that in invasive breast cancer tissue, with no statistically significant difference ( t=-13.193, P>0.999). The median survival time of breast cancer patients in Cyclin D1 high expression group was 173.2 months, which was shorter than 228.9 months in low expression group ( P<0.001). The upper quartile survival time of breast cancer patients in DKK1 high expression group was 55.1 months, which was longer than 40.4 months in low expression group ( P<0.001). The breast invasive ductal carcinoma and breast intraductal carcinoma were combined into tumor group, the sum of the immunohistochemistry scores of β-catenin and Cyclin D1 minus the immunohistochemistry score of DKK1 was used as the combined scoring scheme 1, and the sum of β-catenin and Cyclin D1 immunohistochemistry score was used as the combined scoring scheme 2. ROC curve analysis showed that the area under the curve (AUC) of β-catenin, Cyclin D1, combined scoring scheme 1 and combined scoring scheme 2 for pathologically aided diagnosis of breast cancer were 0.65 ( P=0.080), 0.81 ( P<0.001), 0.70 ( P=0.023) and 0.78 ( P=0.001), respectively. The AUC of Cyclin D1 and combined scoring scheme 2 were ≥0.7, which had good value in pathologically aided diagnosis. Conclusion:Wnt signaling pathway related molecules Cyclin D1 and Cyclin D1 combined with β-catenin detection has a good value in the pathologically aided diagnosis of breast cancer.
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Objective The role of this essay is to explore the relationship of BIM deletion polymorphism and paclitaxel intrinsic resistance in breast cancer. Methods Human breast cancer cell lines were screened by techniques of PCR and gene sequencing to detect BIM deletion polymorphism.MTS was used to detect the cytotoxic effects of paclitaxel in different cell lines. Meanwhile,levels of BIM mRNA and protein were detected by rtPCR and Western Blot. Results Comparing with the wild type cell line(MCF7),T47D was a deletion cell line with BIM deletion polymorphism and resistant to paclitaxel(T47D vs.MCF-7,IC50>30le vs.IC50=0.16 vs.02 μmol/L). Moreover,the ratio of BIM EXON3 to EXON4 was significantly increased and the level of functional BIM protein was down-regulated in T47D compared with MCF7(P < 0.05). Conclusion BIM deletion polymorphism might initiate intrinsic resistance to paclitaxel therapy in breast cancer.
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Objective To explore the clinical outcomes of different mild-stimulation in poor ovarian responders in vitro culture/intracytoplasmic injection (IVF/ICSI) to provide the evidence for clinical treatment,and investigate clinical parameters including pre-ovulation rate,rate of oocytes retrieved,fertilization rate,good quality embryo rate,and pregnancy rate per cycle.Methods The clinical date of a total 180 IVF cycles of infertile patients undergone from Jan 2013 to May 2016 in medical reprodution center of Xiang Tan Central Hospital.According to the different methods of controlled ovarian hyperstimulation (COH),the cycles were divided into 3 groups:group A with 60 cycles of early five days using clomiphene,group B with 60 cycles of persistent using clomiphene,and group C with 60 cycles using artificial luteal-phase ovarian stimulation protocol.Results The basic situations in three groups were no significant difference (P >0.05).Compared to groups B and C,the pre-ovalution rate was more in group A (P < O.05),and the rate of oocytes retrieved,fertilization rate,good quality embryo rate,and preganacy rate per cycle were higher in groups B and C (P < 0.05).The rate of oocytes retrieved,fertilization rate,good quality embryo rate,and preganacy rate per cycle were no significant difference among gourps B and C.The good quality embryo rate and preganacy rate per cycle in groups B and C were higher than group A,and group B was higher than group A (P < 0.05).Conclusions Persistent using clomiphene or persistent using clomiphene in COH could decrease the rate of pre-ovalution,and get more rate of oocytes retrieved,good quality embryo rate,and preganacy rate per cycle.
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Objective To determine the changes in the prevalence of type 2 diabetes mellitus and prediabetes in residents aged over 20 years in Shandong coastal areas during years 2004-2014. Methods A random stratified cluster sampling was conducted, and 3 944 inhabitants were investigated in Qingdao, Yantai, Weihai, and Rizhao regions. Results (1)The standardized rate of diabetes mellitus prevalence was 10.36% in 2014, increased by 2.38% as compared with 2004 and 0.85% as compared with 2009 (P<0.05). There were 341 newly diagnosed diabetes mellitus patients, accounting for 57.4% of the diabetes mellitus patients, increased by 6.09% as compared with 2004 and decreased by 4.11% as compared with 2009 (P<0.05). The standardized rate of prediabetes was 12.47% in 2014, increased by 2.63% as compared with 2004 and 0.61% as compared with 2009 (P<0.05). (2) The prevalence of diabetes mellitus and impaired glucose regulation in individuals aged under 50 years in 2014 was significantly higher than that in 2004, but lower than that in 2009 (P<0.05). (3) Compared with 2004 and 2009, the levels of diastolic blood pressure, total cholesterol, low density lipoprotein-cholesterol (LDL-C), and high density lipoprotein-cholesterol in subjects with prediabetes and diabetes mellitus increased significantly in 2014 (all P<0.05). The levels of serum uric acid in subjects with prediabetes and diabetes mellitus decreased significantly in 2014 (P<0.05). (4) The logistic regression analysis showed that the age, body mass index, waist-hip ratio, triglyceride, LDL-C, systolic blood pressure, and diabetic family history were the risk factors for abnormal glucose regulation, while total cholesterol was the protective factor. Conclusions The prevalence of diabetes mellitus and prediabetes increased during 10 years (2004-2014), but the growth rate dropped precipitously during last five years as compared with the first five years. The age of diabetes mellitus onset is getting older in residents of Shandong coastal area, especially in females. Aging, overweight, hypertension, lipid disorders, and diabetic family history were the risk factors of diabetes mellitus.
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Objective To observe the characteristics of spectral-domain optical coherence tomography (SD-OCT) for leakage point in acute central serous chorioretinopathy (CSC).Methods A total of 21 acute CSC patients (21 eyes) were enrolled in this retrospective study,including 17 men (17 eyes) and 4 women (5 eyes).The mean age was (47.3±8.8) years (range 35-66 years).The mean duration was (1.6±0.8) months (range 0.5-3.0 months).All patients were underwent mydriatic fundus photography,SD-OCT examination and fluorescein fundus angiography (FFA).SD-OCT and FFA images were carefully compared to observe the SD-OCT examination characteristics of fluorescence leakage point.Results 21/21 eyes had one fluorescein leakage point.In addition to serous retinal detachment,leakage point in the SD-OCT examination showed retinal pigment epithelium (RPE) protrusion in 10 eyes (47.6 %),RPE detachment in 7 eyes (33.3 %),highly reflective areas suggesting fibrinous exudate in the subretinal space in 3 eyes (14.3 %),and RPE defect in 1 eye (4.8 %).Conclusion The SD-OCT characteristics of acute CSC include RPE protrusion,RPE detachment,highly reflective areas suggesting fibrinous exudate in the subretinal space and RPE defect.
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Objective To investigate the expression and its potential effect on the immune function of indoleamine 2 ,3-dioxygenase (IDO) in cutaneous malignant melanoma (MM). Methods The IDO protein expression in MM tissue from 30 patients,normal skin from 10 of these patients and normal foreskin of 10 healthy subjects were observed by skin immunohistochemical staining. The number of IDO positive cells were counted. Epidermal cells of MM tissue were isolated and cultured with Hanks solution contained 100 μmol/L tryptophan. The concentration of tryptophan metabolite and kynurenine were detected in supernatant ,reflecting the ability to metabolize the tryptophan of the skin cells. Further ,the isolated epidermal cells were co-cultured with T cells. The influence on PHA-stimulated T cell proliferation were observed. Results IDO(+)cells were almost undetectable in the normal skin,but IDO(+)cells greatly accumulated in epidermis of MM tissue. The percentage of IDO(+) cells/total cells[(28.5 ± 9.4)%]was significantly higher than that of normal skin from healthy subjects[(3.4 ± 1.5)%,P<0.05]and control skin from MM patient[(4.7 ± 2.3)%,P<0.05]. Compared with the epidermal cells from the healthy skin,the IDO(+)cells from MM tissue had a stronger ability to metabolize tryptophan into kyn-urenine in vitro. These isolated IDO-competent epidermal cellscould inhibit PHA-stimulated T cell proliferation. Conclusions IDO(+)cells greatly accumulated in cutaneous MM tissue ,which might induce a local immune tolerant microenvironment.
摘要
Objective To investigate the expression and its potential effect on the immune function of indoleamine 2 ,3-dioxygenase (IDO) in cutaneous malignant melanoma (MM). Methods The IDO protein expression in MM tissue from 30 patients,normal skin from 10 of these patients and normal foreskin of 10 healthy subjects were observed by skin immunohistochemical staining. The number of IDO positive cells were counted. Epidermal cells of MM tissue were isolated and cultured with Hanks solution contained 100 μmol/L tryptophan. The concentration of tryptophan metabolite and kynurenine were detected in supernatant ,reflecting the ability to metabolize the tryptophan of the skin cells. Further ,the isolated epidermal cells were co-cultured with T cells. The influence on PHA-stimulated T cell proliferation were observed. Results IDO(+)cells were almost undetectable in the normal skin,but IDO(+)cells greatly accumulated in epidermis of MM tissue. The percentage of IDO(+) cells/total cells[(28.5 ± 9.4)%]was significantly higher than that of normal skin from healthy subjects[(3.4 ± 1.5)%,P<0.05]and control skin from MM patient[(4.7 ± 2.3)%,P<0.05]. Compared with the epidermal cells from the healthy skin,the IDO(+)cells from MM tissue had a stronger ability to metabolize tryptophan into kyn-urenine in vitro. These isolated IDO-competent epidermal cellscould inhibit PHA-stimulated T cell proliferation. Conclusions IDO(+)cells greatly accumulated in cutaneous MM tissue ,which might induce a local immune tolerant microenvironment.
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The incidence of toxic keratopatby is increasing yearly.Due to the shortage of specific clinical manifestations,the diagnosis and treatment of the disease are relatively difficult,hence toxic keratopathy has received extensive attention of the domestic and abroad scholars.In recent years,with the development of ophthalmic examination technology and progress of instruments,its mechanism,diagnosis and treatment have been newly cognized.These three aspects will be reviewed in this article.
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Objective To investigate the effect of comprehensive intervention on asymptomatic hyperuricemia.Methods in this random controlled study,all patients diagnosed with asymptomatic hyperuricemia on health examination according to the diagnostic standards were randomly divided into intervention group and control group.Comprehensive intervention included healthy lifestyle guidance and administration of sodium bicarbonate in the intervention group.Low purine diet was used in the control group.The follow-up period for all the patients was one year.The changes of various indexes in both groups before and after intervention were observcd and compared.Results Compared with the control group,the patients in the intervention group presented significantly lower levels of BMI,serum uric acid,blood sugar,SBP,DBP,triglyceride and less frequent occurrence of obesity,hyperglycemi,hypertension,hipertrigliceridemia (P < 0.05).After intervention,the patients in the intervention group also exhibited significantly lower the levels of BMI,serum uric acid,blood sugar,SBP,DBP,triglyceride and significantly lower incidences of obesity,hyperglycemia,hypertension,hypercholesteremia and hipertrigliceridemia.Conclusion Comprehensive intervention into asymptomatic hyperuricemia can reduce the incidence of complications such as central obesity,hyperglycemia,hyperlipidemia and hypertension.
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OBJECTIVE To explore the genetic mechanism for a family affected with cardiac conduction block. METHODS Affected family members were screened for potential mutations of known candidate genes. As no pathogenic mutation was found, two patients and one healthy member from the family were further analyzed by exomic sequencing followed by Sanger sequencing. The pathogenicity of suspected mutation was analyzed using bioinformatics software. RESULTS Sequencing of the full exome has identified a c.G1725T mutation in the CLCA2 gene. Sanger sequencing has detected the same mutation in all five patients, but not in the normal member from the family. Bioinformatics analysis indicated that the mutation has resulted in substitution of the 575th amino acid cysteine (C) by tryptophan (W). The site is highly conserved and becomes pathogenic with the mutation. CONCLUSION The heterozygous c.G1725T mutation in exon 11 of the CLCA2 gene probably underlies the disease and fit the autosomal dominant pattern of inheritance.
Subject(s)
Female , Humans , Male , Middle Aged , Amino Acid Sequence , Chloride Channels , Genetics , Computational Biology , Heart Block , Genetics , Molecular Sequence Data , Mutation摘要
ObjectiveTo investigate the characteristics of EGFR gene mutations among NSCLC patients in south of China and analyze the correlation between mutations and clinical features.Methods Specimens of lung cancer tissues were collected from 185 NSCIC patients in our hospital.DNA was extracted from specimens.Exon 18,19,20 and 21 of EGFR gene were amplified by FQ-PCR to be bi-directional sequenced.ResultsEGFR gene mutations in 62 (33.5%) of 185 NSCLC patients was identified in carcinoma tissues,of which,2cases,41cases,5 cases and 14 cases respectively located at exon 18,exon 19,exon 20 and exon 21.The mutation of Del L747 → P752 (P753S) ( proportion 8.1% ),Del E746 → A750 ( proportion 45.1% ) at exon 19 and L858R ( proportion 22.6% ) at exon 21 were the predominant mutation in 16 kinds of mutations.Four cases of mutation at exon 19 got the different results in bi-directional sequencing.The silent mutation 2361G→A at exon 20 was observed (28.1% ).The mutation rate in women was significantly higher than men (46.2% vs 24.3%,x2 =9.670,P =0.002).Non-smokers had significantly higher mutation rate than smokers (41.4% vs 17.1%,x2 =7.380,P =0.007) ; Adenocarcinoma patients had significantly higher mutation rate than squamous cell carcinoma (38.3% vs 6.3%,x2 =6.426,P =0.011).Clinical stage Ⅲ patients had significantly lower mutation ratethan patients with stage Ⅱ orⅣ ( 10.8% vs 53.8%,x2 =8.026,P =0.003 ;10.8% vs 41.3%,x2 =9.518,P =0.002).No statistically significance correlation was found between the mutation ratio and age.ConclusionsEGFR gene mutation has a close relationship with females,non-smokers and adenocarcinoma.Most mutations occur in exon 19 and 20 among patients in south of China.
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ObjectiveTo investigate the effects of bone marrow stem cells autologous transplantation after acute myocardial infarction on local mRNA expression of SDF-1,Kit ligand,E-selectin,ICAM-1and VCAM-1.Methods Acute myocardial infarction models were established in 10 swine,which were randomized divided into placebo group (5 swine) and transplantation group (5 swine).After six months,hearts of placebo group and early transplantation group were harvested and mRNA expression of SDF-1,Kit ligand,E-selectin,ICAM-1 and VCAM-1 in infarcted myocardial region was detected.Results The mRNAs of SDF-1,Kit ligand,E-selection,ICAM-1 and VCAM-1 were expressed in the infarcted myocardial region of transplantation group six months after myocardial infarction,but it was not found in the placebo group.ConclusionsBone marrow autologous transplantation after acute myocardial infarction can promote local mRNA expression of SDF-1,Kit ligand,E-selectin,ICAM-1 and VCAM-1,which may be due to the paracrine effects of stem cells.
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Objective To evaluate the effects of time selection and immediate percutneous transluminal coronary angioplasty (PTCA) in refractory unstable angina pectoris.Methods PTCA was performed in 27 RUAP patients,among which 16 underwent emergency PTCA and 11 patients experienced PTCA after controlling with medicine.ECG and recurrence of symptoms were observed immediately and 6 months after PTCA.Results The procedural success rate was 100%.The immediate relieving rate was 96.3%.The recurrence rate of 11 cases within 6 months (18.2%)was significantly lower than that of 16 cases(37.5%).Conclustion The PTCA for RUAP is relatively safe and effective,but PTCA should be performed after situation is unchanged by use of medicine and PTCA shoulde be carried out immediately if the symptoms can not be controlled by medicine.
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Objective To evaluate the safety and efficacy of low-molecular weight heparin on the prevention of acute closure after percutaneous intracavitary coronary angioplasty.Methods 73 patients were randomly assigned to LMWH and SH groups.All patients underwent SPECT or CAG.Myocardial ischemia and hemorrhage complication were analyzed.Results In Group LMWH,no case of hemorrhage complication occurred,and in 3 cases angina occurred within 24h after the operation,which was proved by CAG to be acute closure of target vessel (the incidence rate 8.6%) .In Group SH,gum and subcutaneous hemorrhage occurred to 2 cases within 7h after the operation,hematuria occurred to 2 cases within 12h and one case died from cerebral hemorrhage,which was confirmed by CT (the total hemorrhage incidence 13.2%).Another 4 cases in Group SH suffered from re-occurrence of angina,which was confirmed by CAG to be acute closure of target vessel (10.5%).There was significant difference in hemorrhage complication between the two groups (P0.05)。Conclusion The results suggest that LMWH is safe and effective in the prevention of acute closure after native vessel coronary angioplasty.
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Objective To study the relationship between expression of EGFR and E- cadherin and metastasis of esophageal cancer. Methods The expression of EGFR and E- cadherin in esophageal cancer tissues of 47 patients by LSAB immunohistochemistry method was studied. Results The positive rates of EGFR in distant metastasis group and local lymph node metastasis group were 47.62 % and 56.52 % respectively. They were higher than individual corresponding control groups(27.08 % and 26.32 %)(P