Objective To construct plvx-cyclooxygenase-2(COX-2)-DsRed and establish breast cancer cell line MCF7 which overexpressed COX-2, to explore the effect of COX-2 on breast cancer cell.Methods The full-length COX-2 PCR product was obtained by total COX-2 PCR primers and COX-2 cDNA vector.After the PCR product and lentiviral vector plvx-DsRed-Monomer-N1 were cut simultaneously by restriction enzyme BamH1 and Xholl, they were connected and sequenced, to get lentiviral vector plvx-COX-2-DsRed.After selected by puromycin, overexpressed COX-2 breast cancer cell line MCF7-plvx-COX-2-DsRed was obtained.The stable cell line was verified by real time PCR and Western blot.The differences of proliferation ability between stable cell line and normal one were compared by colony formation test and Western blot.Results The lentiviral vector plvx-COX-2-DsRed and stable cell line MCF7-plvx-COX-2-DsRed after selecting were obtained.COX-2 expression level of the stable cell line was 75.29 times as high as that of MCF7, and 64.91 times as high as that of cell line MCF7-plvx-DsRed-Monomer-N1 by PCR assay (P ＜ 0.05), which was consistent with the results of Western blot and microscope photo.MTT results showed that cell line MCF7-plvx-COX-2-DsRed had grown faster than cell line MCF7 and MCF7-plvx-DsRed-Monomer-N1 from the 2nd day (P ＜ 0.05), which was accordant with colony formation assay.MCF7-plvx-COX-2-DsRed cell line had higher c-myc expression and lower β-catenin expression than MCF7 cell and cell line MCF7-plvx-DsRed-Monomer-N1 detected by Western blot relative quantification (P ＜ 0.05).Conclusion The plvx-COX-2-DsRed lentiviral vector and cell line MCF7-plvx-COX-2-DsRed are successfully constructed.COX-2 can increase proliferation of MCF7 cells through up-regulating the expression of c-myc.

Objectives To assess the plasma levels of acyl ghrelin (AG) and dys-acyl ghrelin (DG) in chronic kidney disease (CKD) and hemodialysis (HD) patients and analyze their relationships with different stages of CKD and hemodialysis. Methods Forty-six CKD stage 1-5 patients and 15 hemodialysis patients were enrolled into the study. Body weight, height, hemoglobin, biochemical parameters, inflammatory parameters, preprandial, postprandial and 3 hours after hemodialysis plasma AG and DG levels were measured. Appetite and food intake were assessed. Body mass index (BMI), and estimated glomerular filtration rate (eGFR) were calculated. Results There were no significant differences in BMI, SGA, appetite, food intake and malnutrition among CKD patients of different stages. eGFR was declining with the progression of CKD stages and patients received a three-week hemodi-alysis. Compared with that in CKD stage 1-2 patients, the level of preprandial and postprandial DG was remarkably increased in stage 3-5 patients (P<0.01). The level of DG was significantly decreased after a standard breakfast in CKD patients (P<0.01). CKD stage was positively correlated with preprandial (r=0.31, P<0.05)a nd postprandi-al DG (r=0.34, P < 0.05), TNF-α (r=0.33, P < 0.05), IL-6 (r=0.40, P < 0.05), leptin (r=0.34, P < 0.05), and age (r=0.41, P<0.05). CKD stage was also highly and positively correlated with the proportion of preprandial and postprandial DG (r=0.61, P<0.01;r=0.55, P<0.01). Multivariate partial-correlation analysis showed that CKD was independently associated with the proportion of preprandial and postprandial DG (r=0.55, P < 0.01; r=0.43, P < 0.01).There was no decrease in AG postprandially, nor any changes in AG resultant from dialysis (P > 0.05);levels of DG decreased slightly postprandially and were markedly decreased by hemodialysis (P<0.01), even lower than those seen postprandially in CKD stage 1-2;Both preprandial and postprandial DG were negatively correlated with serum albumin levels (r=-0.64, P < 0.05; r=-0.59, P < 0.05), while there was no correlation between AG and serum albumin levels. Conclusions There is a strong and independent correlation of DG with CKD stage. Postprandial suppression of ghrelin is impaired with reduced renal function. Hemodialysis removes DG but not AG.

Objective To design an interactive and shared electronic database for long-term follow-up management of patients with acute minor stroke (NIHSS≤5) using REDCap,and to explore the value of establishing this database,trying to provide new ideas for clinical treatment. Methods The CRF table of case report was designed according to the relevant data of patients in hospital and the requirements of follow-up management. The nosocomial case data of patients with acute minor stroke (NIHSS≤5) from 3 stroke centers in Shanxi Province were collected and recorded on the CRF form of case reports. An interactive shared electronic database was designed by REDCap,and the data in CRF table were checked and revised and entered into the database. Patients were followed up at 3 months and 1 year after onset. Results Based on REDCap system,a database of acute minor stroke ( NIHSS≤5) in Shanxi Province was established and used in clinical practice. The number of patients expected to be included has been achieved. Its data entry,data quality control,user rights management and data export functions can be stable operation. Conclusion The interactive sharing clinical database of acute minor stroke ( NIHSS≤5) is established by redcap,which has the advantages of simple interface operation,convenient communication,timely entry,and multi-access. It provides a powerful tool for longitudinal data collection,reducing deviation in research,and comprehensively implementing and coordinating project research. It ensures the reliability of research results and has clinical research value.

Objective:To investigate occupational hazards in waste incineration power generation enterprises.Methods:From May 2016 to September 2018, four garbage incineration power generation enterprises in different regions and scales were selected as the survey objects. Investigated and analyzed the types of occupational hazards, workplace distribution and occupational disease prevention facilities and the concentration (intensity) of major types of work and workplace occupational hazards were also improved Line detection and analysis.Results:Silica dust could be seen in slag tank, slag sorting and crusher operation position. The concentration of silica dust in each workplace fluctuates within 0.3-1.1 mg/m 3. Both the concentration of silica dust in the workplace and the time weighted average allowable concentration (C-TWA) of dust exposure of dust exposed workers were detected, but none of them exceeds the requirements of the national occupational exposure limit. Carbon monoxide, ammonia, nitrogen dioxide, hydrogen sulfide, non methane total hydrocarbon, formaldehyde, hydrogen fluoride, hydrogen cyanide, hydrochloric acid, sodium hydroxide and ozone were detected, but all meet the requirements of national occupational exposure limit. The 8-hour equivalent a sound level (L ex·8h) over 85 dB (A) mainly includes boiler, steam turbine, electrical inspector and auxiliary value of water treatment. Conclusion:The occupational hazards of waste incineration power generation enterprises are serious. Occupational health management and occupational health monitoring to reduce their occupational hazards to workers.

Objective:To construct a color assessment tool for continuous bladder irrigation (CBI) drainage fluid and to explore its clinical value.Methods:The color assessment tool for CBI drainage fluid (tentative version) was constructed through literature review, expert meetings and practical development, and its consistency was tested and the tool was improved to form a color assessment tool for CBI drainage fluid. Convenience sampling was used to select patients with benign prostatic hyperplasia who underwent transurethral resection of the prostate (TURP) in the First Affiliated Hospital of Soochow University from April to May 2020. According to the assessment method of the tool, the patient's drainage fluid color was evaluated and recorded.Results:The color assessment tool for CBI drainage fluid included 6 color scales, and the corresponding Pantone color card codes were 503C, 3519C, 701C, 702C, 710C, 1807C respectively, and the blood concentration was 0.125%, 0.5%, 1.0%, 2.0%, 4.0% and 8.0% respectively. The tool was used to evaluate the drainage fluid color in 29 patients after TURP. Each evaluation took (5.12±2.41) s; when the color scale was greater than number two, complications such as the bladder spasm and blood clot blockage occurred.Conclusions:The construction method of color assessment tool for CBI drainage fluid is scientific and reliable, which can provide a basis for evaluating the CBI drainage fluid color, and can assist medical and nursing staff to quickly evaluate the drainage fluid color and find abnormal color scales.

Objective:To investigate occupational hazards in waste incineration power generation enterprises.Methods:From May 2016 to September 2018, four garbage incineration power generation enterprises in different regions and scales were selected as the survey objects. Investigated and analyzed the types of occupational hazards, workplace distribution and occupational disease prevention facilities and the concentration (intensity) of major types of work and workplace occupational hazards were also improved Line detection and analysis.Results:Silica dust could be seen in slag tank, slag sorting and crusher operation position. The concentration of silica dust in each workplace fluctuates within 0.3-1.1 mg/m 3. Both the concentration of silica dust in the workplace and the time weighted average allowable concentration (C-TWA) of dust exposure of dust exposed workers were detected, but none of them exceeds the requirements of the national occupational exposure limit. Carbon monoxide, ammonia, nitrogen dioxide, hydrogen sulfide, non methane total hydrocarbon, formaldehyde, hydrogen fluoride, hydrogen cyanide, hydrochloric acid, sodium hydroxide and ozone were detected, but all meet the requirements of national occupational exposure limit. The 8-hour equivalent a sound level (L ex·8h) over 85 dB (A) mainly includes boiler, steam turbine, electrical inspector and auxiliary value of water treatment. Conclusion:The occupational hazards of waste incineration power generation enterprises are serious. Occupational health management and occupational health monitoring to reduce their occupational hazards to workers.

Objective:To investigate the relationship between plasma fluoride content, daily calcium intake and blood cell parameters in children and adolescents.Methods:This study was based on the National Health and Nutrition Examination Survey (NHANES) database of the United States from 2013 to 2016, with 3 684 children and adolescents aged 6 - 19 as the research subjects. Information on plasma fluoride content, daily calcium intake and blood cell parameters from the database were collected. Non-linear relationships between plasma fluoride content, daily calcium intake and blood cell parameters were analyzed using restricted cubic splines. If there was a non-linear relationship, the optimal inflection point was calculated using threshold/saturation effect analysis method. Subsequently, multiple linear regression models were used to analyze the associations among the three, and the modification effect of daily calcium intake (binary classification, stratified by median daily calcium intake) on the association between plasma fluoride content and blood cell parameters was analyzed.Results:There was no non-linear relationship between plasma fluoride content and white blood cell count, hemoglobin content and platelet count ( Pnon-linear > 0.05), but there was a non-linear relationship between plasma fluoride content and erythrocyte count and hematocrit ( Pnon-linear < 0.001). After adjusting for confounding factors, the optimal inflection points of the effects of plasma fluoride content on erythrocyte count and hematocrit were 0.54 and 0.31 μmol/L, respectively. There was no non-linear relationship between daily calcium intake and blood cell parameters ( Pnon-linear > 0.05). After adjusting for confounding factors, for every 1 μmol/L increase in plasma fluoride content, the white blood cell count increased by 0.49 × 10 9/L ( P = 0.009). There was a saturation effect in the association between plasma fluoride content, erythrocyte count and hematocrit: when plasma fluoride content was < 0.54 μmol/L, the erythrocyte count decreased by 0.46 × 10 12/L for every 1 μmol/L increase ( P < 0.001). When plasma fluoride content was < 0.31 μmol/L, the hematocrit decreased by 6.29% for every 1 μmol/L increase ( P = 0.006). The above associations were not statistically significant when plasma fluoride content was higher than the optimal inflection points ( P > 0.05). After stratification according to the median daily calcium intake, in the low-calcium group (daily calcium intake < 0.87 g), for every 1 μmol/L increase in plasma fluoride content, the white blood cell count increased by 0.77 × 10 9/L ( P = 0.001). When plasma fluoride content was < 0.54 μmol/L, the erythrocyte count decreased by 0.41 × 10 12/L for every 1 μmol/L increase ( P = 0.002). When plasma fluoride content was ≥0.54 μmol/L, erythrocyte count decreased by 0.47 × 10 12/L for every 1 μmol/L increase ( P < 0.001). When the plasma fluoride content was < 0.31 μmol/L, the hematocrit decreased by 8.29% for every 1 μmol/L increase ( P = 0.011). The above associations were not statistically significant in the high-calcium group (daily calcium intake ≥0.87 g, P > 0.05). There was an interaction of daily calcium intake and plasma fluoride content on platelet count ( Pinteraction = 0.070), as demonstrated by an increase in platelet count of 12.68 × 10 9/L ( P = 0.013) in the low-calcium group and a decrease in platelet count of 9.05 × 10 9/L ( P = 0.035) in the high-calcium group for every 1 μmol/L increase in plasma fluoride content. Conclusions:The blood cell parameters of children and adolescents are closely related to plasma fluoride content, but not directly related to daily calcium intake. However, the correlation between plasma fluoride content and blood cell parameters varies among different calcium intake populations, and daily calcium intake can modify the association between plasma fluoride content and platelet count.

Objective:To observe the morphological and hemodynamics changes of aortic segments in mice with angiotensinogen Ⅱ(Ang II) combined with β-aminopropionitrile(BAPN) induced-aortic dissection by color Doppler ultrasound(CDUS).Methods:Twenty male mice of 6-8 weeks old C57BL/6 were randomly divided into two groups: the model group( n=10) was induced by intraperitoneal injection of Ang Ⅱ combined with BAPN to establish mice model with aortic dissection; the control group( n=10) was intraperitoneally injected with normal saline.The body weight, systolic and diastolic blood pressure of the mice were routinely recorded. On the 42th day, CDUS was used to measure the indexes of ascending aorta(AoA), descending thoracic aorta(DAo) and suprarenal aorta(SAo) in both groups, including the inner diameter of the cross section, peak systolic velocity(PSV), the end diastolic velocity(EDV), the resistance index(RI), the pulsatility index(PI), time average mean velocity(TAMV), the heart rate(HR) and the maximal shear rate(SR). Then, the aortas were harvested from the root to the bifurcation of the renal artery. The pathological changes of the aortic wall were observed using hematoxylin-eosin(HE) staining. Results:①There were statistically significant differences in body weight, systolic blood pressure, diastolic blood pressure and heart rate between the model group and the control group(all P<0.05). Compared with the control group(0/10), the incidence of the AoA dissection(8/10) in the model group was obviously higher, the difference was statistically significant( P<0.05); while the incidence of the DAo dissection(4/10) and the SAo dissection(3/10) in the model group was slightly higher, the differences were not statistically significant (all P>0.05). ②Compared with the ascending aorta of the control group, the inner diameter, PSV, EDV, TAMV, PI and SR in the model group were significantly higher(all P<0.05), while RI showed no significant difference between the two groups ( P>0.05). For the descending thoracic aorta, PSV, EDV, TAMV, PI and SR in model group were higher than those of the control group(all P<0.05), however the inner diameter and RI were not significantly different between the two groups (all P>0.05). And for the superior renal aorta, PSV, TAMV, RI, PI and SR in the model group were obviously higher than the control group(all P<0.05), whereas the inner diameter and EDV were not significantly different between the two groups (all P>0.05). ③The HE of the tissue section in the model group showed, the aortas were obviously dilated, irregular, with inhomogeneously thickening wall; the endothelial cell nuclei were slightly stained, and some intima and middle layer ruptured and protruded outward to form dissecting aneurysms. The adventitias were markedly infiltrated with inflammatory cells. Conclusions:Ultrasonography could primarily evaluate the hemodynamic changes of aorta in hypertension with aortic dissection, and the PSV, TAMV, PI and SR of aorta may be important indicators for early predicting the occurrence of aortic dissection in hypertension.

Objective To characterise the alterations of serum autoantibodies for cyclinB1,p62,Koc-IMP1 and survivin in the subjects with esophgeal and gastric cardia carcinoma and precancerous lesion and their expres-sions in the esophageal and gastric cardia cancer tissue.Methods Enzyme-linked immunoassay and tumor-associated antigen mini-array (consisting of five full-length recombinant proteins,including eyefinB1-p62-Koc,IMP1 and Survivin)were applied to determine the serum level of the autoantibodies of these antigens on 376 subjects with e-sephageal and gastric cardia carcinoma and precancerous lesions.At the same time,the expression of these antigens was detected by immunohistochemical method(ABC)on 13 patients with esophageal cancer and 16 with gastric car-dia cancer.Results All of the 5 antigens determined,the linear correlation Was observed for the detection frequency of cyclinB1,IMPI and p62 in esophageal carcinogenesis,and for p62 in gastric cardia multi-stage progression from normal to precancerous and cancerous lesions(P<0.05).The detection rale with single positive antoantibody im-munoreactivity for both esophageal and gastric cardia cancers were low.However.the positive detection mte for both esophageal and gastric cardia cancer increased apparently when the multiple positive markers were combined together for analysis,which increased tO 3～5 and 3～4 folds respectively.Furthermore,the difference in autoantibody immu-noreactive rate was significant with the lesion progressed from mild tO severe precancerous lesions and to cancer both in esophageal and gastric cardia cancers(P<0.05).The positive immunoreactions of the 5 antigens were detected in cancer tissues.The positive immunostaining rates for cyclinB1,Koc,IMP1 and Survivin both in esophageal and gastric cardia cancers were higher compared to their serllin positive rate of autoantibodies I P<0.05).Of the pa-tients with positive immunostaining in the two cancer tissues,the autoantibodies in the serum for the corresponding antigens could be detected in the same patient.Conclusion The production of the tumor-associated autoantibodies is related tO antigens.The screening rate through serum tumor-associated antigen mini-array for the patients with e-sophageal and gastric cardia carcinoma and precancerous lesions has been increased apparently with combined analy-sis of multiple autoantibodies than with single one.

Clostridium difficile infection (CDI) is a growing public health concern worldwide. Antibiotics are currently the preferred treatment for CDI, but the heavy use of antibiotics has resulted in the emergence of highly resistant strains of Clostridium difficile and recurrent CDI. More and more studies show that restoring intestinal flora homeostasis is an important strategy for the treatment of CDI. This article reviewed the relationship between intestinal flora homeostasis and CDI, and the treatment strategies for CDI by restoring intestinal flora homeostasis.