Objective To study the effects of different levels estradiol on inhibitory of tamoxifen on human mammary cancer cells(ER+) in vitro.Methods Estrogen receptor(ER)-positive MCF7 human breast cancer cell line was treated by the same level of tamoxifen and different levels of estradiol in vitro.Cell proliferation was evaluated by MTT assay.Results E_2 at concentrations between 1? 10~(-12) mol/L to 1? 10~(-7) mol / L significantly stimulated the growth of MCF-7.TAM(10 ?mol/ L) inhabited the growth of MCF-7 significantly.E_2 at different levels may influence inhibitory of tamoxifen on MCF-7 cell lines.E_2(1?10~(-8) mol/L) makes inhibitory of tamoxifen on MCF-7 cell lines valueless.Conclusion E_2 is the risk factor of breast cancer,and the concentration around breast cancer cells may influence the effects of TAM.

Objective To study the expression and significance of miR-26a in intrahepatic cholangiocarcinoma.Methods The expression of miR-26a in 46 intrahepatic cholangiocarcinoma(ICC) tissues and peritumoral tissues was detected by quantitative real time polymerase chain reaction (qRT PCR).The intrahepatic eholangiocarcinoma cell line HCCC-9810 and RBE were transfected with miR 26a mimics and miR 26a inhibitors,respectively,by lipofectamine 2000.The growth curves were constructed by the CCK 8 method.The migration and invasion ability was demonstrated by wound healing and transwell assay.The cell cycle was analyzed by flow cytometry.The potential mechanism was illustrated by Western blotting.Results For the 46 ICC tissues and peritumoral tissues,miR 26a levels were significantly higher in the tumor tissues than in the peritumoral tissues (P＜0.05).Vascular invasion,TNM Ⅲ～Ⅳ stage and lymph node metastasis were significantly associated with high miR 26a expression levels (P＜0.05),but gender,age,tumor amounts,tumor encapsulation,tumor diameter and tumor differentiation showed no significant association (P＞0.05).Enhanced cell proliferation,migration and invasion ability,accelerated G0/G1 phase to S phase transition,activated AKT by PTEN suppression were observed in HCCC-9810 cells with up regulation of miR-26a.Conversely,cell proliferation,migration and invasion ability was inhibited,G0/G1 phase was blocked and AKT was restrained by PTEN increase wkh down regulation of miR-26a in RBE cells.PTEN mRNA in versely correlated with the miR-26a level (r=-0.8272,P＜0.01).Patients with a high miR-26a expression had significantly poorer overall survival (P＜0.05).A high miR 26a exprcssion,multiple tumors and lymph node metastasis were independent prognostic factors of overall survival (P＜0.01).Conclusion Overexpression of miR-26a in intrahepatic cholangiocarcinoma correlated with clinicopath ological features and overall survival,and it potentially promoted tumor proliferation and metastasis through the PTEN/AKT pathway.

Objective To understand the effect of estradiol in different concentrations on proliferation of diverse mammary primary cells in vitro. Methods The primary cells of cancer tissue, the adjacent tissue to tumors and normal mammary tissue from patiens with breast cancer were obtained using collagenase digesting method. All the tissue samples were cultivated in vitro, and were given estradiol in different concentrations. The effect of estradiol on the proliferation of those primary cells was measured by MTT. Results Estradiol remarkedly promoted the proliferation of primary cells of cancer tissue and peritumor tissue in vitro, whose ER expression were positive. Whereas, the promotion effect of estradiol on the proliferation of normal mammary primary cells was relatively weak, and there was no correlation between the promotion effect with the expression of ER in cancer tissue. Conclusion The risks of occurrence and relapse of breast cancer would increase significantly when the concentration of estradiol is no less than 103 pmol/L in vivo.

Objective: To observe the influence of transcutaneous CO2emission (TCE) at points on the meridian or beside the meridian after different stimulations like acupuncture De-qi, sham acupuncture, study the correlation between acupuncture De-qi and local energy metabolism through TCE. Methods: Twenty healthy volunteers (male 8, female 12, average age 29 years old) were selected. Before, during and after five different acupuncture Neignan (PC 6), the TCE was measured at Quze (PC 3), point A [at the lower one fourth of line between Tianquan (PC 2) and Quze (PC 3)], point B [at the middle of line between Ximen (PC 4) and Quze (PC 3)], point C (2 cm ulnar side point A) and point D (2 cm ulnar side point B) with a high sensitive C02 instrument (Frewil-Q.F., made in China). The measure results were compared. Results: The comparison on value of TCE before and after the acupuncture, in Sham acupuncture and shallow acupuncture groups, it increased in all five points; in De-qi acupuncture group, there are significant increase on point A, point B, and Quze (PC 3), it has statistic significance, but there are no significant increase on point C and D. After Ximen (PC 4) or Daling (PC 7) were pressed to block Neiguan (PC 6), the value of TCE on Quze (PC 3), point A and point B were decreased during acupuncture Neiguan (PC 6), and increased after the block was removed, but there was no significant difference. Conclusion: Sham acupuncture has also some effects on body, it can increase energy metabolism of local skin, it may be the reason that shallow acupuncture without qi amving can obtain clinical efficacy. The operation method of traditional qi arriving acupuncture can increase energy metabolism of relevant meridian skin, mechanical press can reduce this effect.

Background Apoptosis is a primary clinical pathological mechanism of diabetic retinopathy (DR).Oxidative stress and high glucose can activate cell apoptosis pathway and thus leads to cellular damage.It is confirmed that tert-butyl hydroquinone (tBHQ) plays an antioxidation effect,however,whether it has a protective role on retinal cells in DR is still unelucidated.Objective This study was to investigate the effect of tBHQ on vascular endothelial growth factor (VEGF) and bcl-2 expressions in retina of type 2 diabetic rats and its possible mechanism via nuclear factor erythroid 2-related factor 2/antioxidant response element (Nrf2/ARE) signal pathway.Methods Fifty clean healthy male SD rats were included in this experimental study.Ten rats were fed with normal diet as the normal control group,and other rats were fed with high fatty and high sugar food for 4 weeks.After 12 hours of fasting,streptozotoin (STZ) (30 mg/kg) was intraperitoneally injected to induce the type 2 diabetic models.The model rats were randomly divided into the diabetic control group and tBHQ group and 1％ tBHQ was added into the high fatty and sugar food 1 week after modeling in the tBHQ group.Fasting plasma glucose (FPG) level,blood total cholesterol (TC) level,blood triglyceride (TG) level,high density lipoprotein-cholesterol (HDL-C),low density lipoproteincholesterol (LDL-C) and fasting serum insulin (FINs) were detected 4 and 12 weeks after modeling,respectively,and radio immunoassay was used to detect the FIN levels of the rats.The relative expression of VEGF and bcl-2 in retinas of the rats were assayed by immunohistochemistry and fluorescence real-time quantitative PCR (qRT-PCR).The use of the animals complied with the Regulations for the Administration of Affairs Concerning Experimental Animals by State and Technology Commission.Results Type 2 diabetic models were successfully established in 35 rats with successful rate 92.1％.The FIN levels were significantly different among different groups and time points (Fgroup =22.480,P =0.000;Ftime =7.636,P =0.008).The FPG,TC,TG and LDL-C levels were significantly different among the groups (FPG:Fgroup =78.531,P =0.000;TC:Fgroup =28.049,P =0.000;TG:Fgroup =13.108,P =0.000;LDL-C:Fgroup =6.804,P＜0.05).Immunohistochemistry showed that VEGF and bcl-2 were mainly expressed in retinal ganglion cell layer,inner plexiform layer and outer plexiform layer.The expressions of VEGF and bcl-2 proteins were significantly different among different groups (VEGF:Fgroup =11.805,P =0.000;bcl-2:Fgroup =22.943,P =0.000);the expression level of bcl-2 protein was higher in 12 weeks after modeling than that in 4 weeks after modeling in the tBHQ group (P＜0.05).The expressions of VEGF and Bcl-2 mRNA in rat retinas were significantly different among different groups and time points (VEGF:Fgroup =79.220,P =0.000;Ftimo =6.090,P＜0.05;Bcl-2:Fgroup =105.000,P=0.000;Ftime =13.170,P=0.001).Four and eight weeks after modeling,the expressions of VEGF and Bcl-2 mRNA in the diabetic control group and tBHQ group were significantly higher than that in the normal control group,and the expressions of Bcl-2 mRNA in the tBHQ group were significantly higher than that in the model control group (all at P＜0.05);the expression of Bcl-2 mRNA was higher at 12 weeks after modeling than that at 4 weeks in the tBHQ group (P＜0.05).Conclusions tBHQ produces anti-oxidative-damage and anti-apoptosis effects on retinal cells by up-regulating VEGF expression and down-regulating bcl-2 expression in DR rats.In addition,tBHQ may have effects on lowering high blood sugar,regulating insulin and blood lipid levels.

OBJECTIVETo compare the difference in the efficacy on low back pain of cold-damp pattern between electric-thermal Bian-stone therapy and moxibustion box therapy.

METHODSForty-one cases of low back pain of cold-damp pattern were randomized into an electric-thermal Bian-stone therapy group (group A, 26 cases) and a box moxibustion therapy group (group B, 15 cases). In the group A, the electric-thermal Bian-stone was placed over Shenshu (BL 23) and Weizhong (BL 40). The temperature of stone was adjusted in accordance with patient's comfort. In the group B, moxibustion box was used over Shenshu (BL 23) and Weizhong (BL 40). The treatment was given once every day or every two days. Ten treatments made one session. The symptom and physical signs score of low back pain and the score of cold-damp syndrome were observed before and after treatment in the patients.

RESULTSThe symptom and physical signs score of low back pain and the score of cold-damp syndrome were all improved in the two groups (all P < 0.01). The changing rate of symptom and physical signs and syndrome were (37.04 +/- 32.68)% and (22.85 +/- 29.95)% in the group A, and were (47.29 +/- 22.08)% and (23.89 +/- 22.53)% in the group B, respectively, without significant difference in comparison between the two groups (all P > 0.05).

CONCLUSIONThe efficacy on low back pain of cold-damp pattern treated with the electric-thermal Bian-stone therapy is similar to that of moxibustion box therapy. This therapy is characterized as more convenient, safer operation and less pollution.

Acupuncture Points ; Adult ; Aged ; Aged, 80 and over ; Combined Modality Therapy ; Female ; Humans ; Low Back Pain ; therapy ; Male ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Moxibustion

Objective To optimize several key factors in preparing protein microarray,and establish microarrays to detect autoantibodies.Methods Four factors,including incubation time of the microarray after spotting,the blocking reagent,the length of blocking time,the length of time conjugating with the second-an- tibody,were selected to carry out orthogonal optimization,then we determined the optimal spotting concentra- tion of antigen and the best dilution of serum with the optimized conditions obtained from the above.On the basis of the optimized conditions,we prepared microarray to detect six autoantibodies simultaneously,which were compared with IIF and ELISA.Results The best conditions we determined from the experiment were: incubating the spot microarray for 120 min,PBST containing 1% BSA and 2.5% saccharose as the blocking reagent and blocking for 30 min,reacting with the second-antibody for 30 min,the appropriate spotting con- centrations of six antigens differed from 100?g/ml to 300?g/ml and the best serum dilution was 1:2.Com- pared with IIF for ANA detection,the sensitivity of the microarray was 88.6%,the specificity was 83.3%,and compared with ELISA in detecting the other five autoantibodies,the sensitivity of the microarray was between 80.0% and 88.2%,and the specificity was between 90.2% and 98.6%.Conclusion The optimization condi- tions we obtained are suitable for preparing protein microarray,and the detection sensitivity and specificity of autoantibodies by microarray are consistent with the conventional methods in general.The microarray for de- tecting autoantibodies is applicable in the clinical diagnosis of autoimmune disease.

Objective To summarize the experience about the diagnosis and treatment of bladder benign neoplasm in children.Methods A retrospective study was conducted for a total of 15 patients with bladder benign neoplasm from October 2006 to May 2016.There were 10 males and 5 females with a mean age of 8.7 years (ranging 1.1-13.8 years).The clinical manifestations of 15 patients included hematuria in 9 patients,frequent micturition with urgent and painful in 3 patients,dysuria in 1 patient,abdominal pain in 2 patients and headache during voiding in 1 patient.Ultrasound showed solid masses in the bladder with iso-echoic or nonhomogeneously hypoechoic.CT scanning showed regular or irregular mass with some enhancement in the bladder.All cases received tumor complete resection by opening operation and bladders were preserved.Among the 15 cases,neoplasms located in the anterior,posterior and lateral wall of bladder in 9 cases,ureteral orifice in 4 cases and trigone of bladder in 2 cases.The size of tumors ranged from 1.2 to 6.0 cm (mean 3.1 cm).The tumors were unifocal and seemed like papillary or cauliflower.The literatures of benign neoplasm of bladder were reviewed,which focused on the clinic characters,pathological classification and therapeutic method.Results Pathologic type included papilloma in 5 patients,inverted papilloma in 1 patient,inflammatory myofibroblastic tumor in 7 patients,hemangioma in 1 patient,pheochromocytoma in 1 patient.Fifteen patients were followed up for 6 to 36 months,mean 26.7 months.All patients recovered well without relapse or metastasis Conclusions Bladder benign neoplasm in children is rare with many kinds of pathological classification.The major clinical manifestation is gross hematuria while dysuria is unusual.Tumors are fewer in trigone of bladder.The best treatment is to resect the tumor completely with bladder preservation if possible.

Objective:To evaluate the assistant role of manifestations under tracheoscopy in the diagnosis of invasive pulmonary aspergillosis (IPA) in severe patients.Methods:A retrospective study was conducted. The patients with suspected IPA admitted to intensive care unit (ICU) of Affiliated Hospital of Binzhou Medical College from January 2015 to December 2019 were enrolled. The diagnosis, clinical diagnosis and suspected diagnosis were made according to the grading criteria of Guidelines for the diagnosis and treatment of invasive fungal infection in severe patients (2007). Those who met the criteria were enrolled in the IPA group, and those who did not meet the criteria or other pathogens were enrolled in the non-IPA group. The general data of the patients were collected, and the changes of tracheal and bronchial mucosa under tracheal microscope before and after treatment were recorded, as well as the results of galactomannan (GM) test and aetiology culture of bronchoalveolar lavage fluid (BALF). The baseline, bronchoscopy and pulmonary CT manifestations and their dynamic changes were compared in each group. Results:A total of 142 patients with suspected IPA were finally enrolled. Among them, 12 were pathologically proven IPA, 77 were probable IPA, 22 were possible IPA, and 31 were undefined IPA. Of the 142 patients, 60 had typical manifestations of mucosal injury under bronchoscopy, including 7 proven IPA patients (58.3%), 52 probable IPA patients (67.5%), and 1 possible IPA patient (4.5%), but none undefined IPA patient. The patients undergoing lung CT scan were 12 proven IPA patients (100%), 73 probable IPA patients (94.8%), and 21 possible IPA patients (95.5%), respectively. Most of the Chest CT showed patchy or strip density increasing and other non-specific manifestations. There were 3 proven IPA patients (25.0%), 7 probable IPA patients (9.0%), and 0 possible IPA patient (0%) who had typical IPA CT manifestations (halo sign and cavity or crescent sign). Among the patients of proven IPA and probable IPA (89 cases), there were a total of 35 cases with endoscopic airway mucosal injury and tracheoscopy reexamination ≥ 3 times. All the 35 patients received anti-aspergillus treatment, among which 16 survived and 19 died. Among the 16 patients who survived, the microscopic appearance of mucosal injury was gradually reduced and the clinical manifestations were gradually improved. Of the 19 patients who died, 16 had deteriorated endoscopic airway mucosal injury.Conclusions:The specific manifestations of severe patients with bronchial mucosal injury are of great significance in the diagnosis of IPA. In the case of severe patients who cannot receive pathological examination or chest CT in time, dynamic observation of the changes of airway mucosal injury is a simple auxiliary method to discover the changes of patients' condition in time, evaluate the effect of antifungal therapy and the prognosis of IPA.

Objective To observe the anatomic and imaging morphology ofjugnlar bulb and its relationship with the surrounding structures, and to investigate the classification ofjugnlar bulb and its clinical significance. Methods We dissected 30 human temporal bones and studied multi-slice spiral CT imaging data of temporal bone of 120 cases and blood vessel cast mould specimen of the jugular bulb of 6 cases, to observe the morphology of jugnlar bulb and its spatial relationship with the surrounding structures. We made an imagined sagittal plane on the medial well of the tympanic cavity, with a horizontal tangent line of the proximal wall of the tympanic cavity and a vertical tangent line of the posterior wall of the tympanic cavity as coordinate axes (X axis and Y axis), respectively, so the 4 quadrants ( Ⅰ , Ⅱ, Ⅳ, Ⅳ) were formed. The jugular bulb was classified intro 4 types according to the quadrant where its top was projected and subtyped according to its position on the inner or outer side of the plane. The operation via mastoid approach was simulated on specimen to observe the effect of jugnlar bulb on the operation route. Results Some jugular bulbs were flat type and others were prominent types. The classification in the group of CT image: type Ⅰ , 11 case (9%);type Ⅱ, 63 cases (53%);type Ⅲ, 25 cases (21%);type Ⅳ, 21cases (17%). The classification in the group of specimen: type Ⅰ, 1 case (3%);type Ⅱ, 11 cases (37%);type Ⅲ, 8 cases (27%);type Ⅳ, 10 cases (33%). Each type of the jugular bulb had different effects on the operative approach. Conclusions The classification method with the 4 quadrants is a simple and three-dimensional way to describe the position of the jugular bulb for imaging diagnosis or operative scheme design.