Objective To construct the prokaryotic recombinant expression vector of PTD4-Cu,Zn-SOD.Methods By using the techniques of gene recombination,the primers of Cu,Zn-SOD and the oligonucleotide sequences of PTD4 were designed,PCR amplification was performed for Cu,Zn-SOD genes,the PCR products were identified,reclaimed and purified,and pET16b served as carrier.The prokaryotic recombinant expression vector of pET16b-Cu,Zn-SOD was constructed using double digestion with Xho Ⅰ and BamH Ⅰ,ligated reaction and plasmid transformation.Then PTD4 gene and pET16b-Cu,Zn-SOD carrier were double digested with Nde Ⅰ and Xho Ⅰ and ligated,and the plasmid was transformed,and the prokaryotic recombinant expression vector of pET16b-PTD4-Cu,Zn-SOD was constructed.The reconstructed vector was analyzed by restriction mapping and was verified by gene sequencing.Results The prokaryotic recombinant expression vector of pET16b-PTD4-Cu,Zn-SOD with a length of 6 207 bp was constructed successfully.The carrier fragment about 5.7 kp and PTD4-Cu,Zn-SOD gene fragment about 510 bp were obtained by double digestion with Nde Ⅰ and BamH Ⅰ,which was consistent with the expected results.The results of gene sequencing showed that the base sequences of pET16b-PTD4-Cu,Zn-SOD were correct when compared with the expected gene sequences.Conclusion The prokaryotic recombinant expression vector of pET16b-PTD4-Cu,Zn-SOD is constructed successfully.

Objective To compare the value of contrast-enhanced ultrasonography (CEUS) with that of laparoscopic ultrasonography (LUS) in localization diagnosis of insulinoma. Methods Thirty-three patients with insulinoma,who received operation, were retrospectively reviewed. All cases were examined by both CEUS and LUS before the tumor removed. Both imaging modalities in diagnosis of insulinoma were analyzed. Results By this study the sensitivity and accuracy of CEUS were 92.31% and 90.0% respectively. Those of LUS were consistent with pathologic findings completely. The accuracy of CEUS was not different with that of LUS (χ2 = 4.21, P = 0. 04). Conclusions CEUS can be used for location diagnosis of insulinoma. For the patients expecting laparoscopic treatment, combination of CEUS with LUS will be suggested before insulinoma is removed.

Objective To investigate the level and influencing factors of PICC catheter patients' willingness to participate in patient safety. Methods A total of 198 PICC catheter patients were sampled from the targeted hospital by convenience sampling,and investigated by the Patient Participate Patient Safety Willingness and Behavior Scale ( PSWBS). Results The PSWBS score of PICC catheter patients was 3.73 ± 0.52 , including initiative willingness 4.13 ± 0.54, initiative tendency 3.82 ± 0.49, attention for technique level 3.70 ± 0.50, attention for environmental safety 3.27 ± 0.48. Among these factors, age(F=14.75, P<0.01), education degree(F=9.588, P<0.01), occupation(25.070, P<0.01), residence(t=8.570, P<0.01)and complications(t=8.880, P<0.01)were the influencing facts with statistical difference (P<0.05) . Conclusions Most of the patients have a high willingness to participate in patient safety , the distribution of impact factors have certain characteristics. Clinical nurses should take targeted nursing measures according to these different characteristics.

Objective To compare the simple cases of hand-foot-mouth disease(HFMD) with HFMD patients complicated with encephalitis and HFMD cases complicated with pulmonary edema (PE). To explore predictor factors of disease progression and unfavorable prognosis. Methods Forty-one EV71-infected children admitted to the Fuyang First People's Hospital in Anhui Province from March to May in 2008 were investigated in the research, who were classified as encephalitis-complicated cases ( encephalitis group, n = 15 ), PE-complicated cases ( PE group, n = 15 ) and simple cases (simple group, n= 11 ). Their clinical manifestation, laboratory findings, and immunophenotypes of peripheral blood lymphocyte were analyzed to find predictors associated with disease progression and unfavorable outcomes. Results The mortality rate in PE group was 66.7%, which was significantly higher than that in encephalitis group. Ninty-three point three percent cases in PE group and encephalitis group were younger than 3 years old, with statistic difference compared to simple group. Patients in PE group had higher total blood white cell (WBC) counts and higher absolute neutrophil counts and tended to have higher breathing rate, heart i'ate and glucose level than encephalitis group. The percentages of T cells and natural killer (NK) cells were significantly lower among patients complicated with encephalitis than simple HFMD patients.Conclusions PE is one predictor for poor prognosis. Factors correlated with unfavorable outcome include high WBC, high absolute neutrophil counts; elevated breathing rate, heart rate and glucose level. The immunophenotypes of peripheral blood lymphocytes can also predict the disease progression.

Objective To investigate the correlation between wholeGorgan MRI score (WORMS)and pain in patients with knee osteoarthritis (OA).Methods 1 1 9 patients with knee OA were enrolled,and The Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC)was used to estimate the pain.Knees of all the subj ects underwent scanning with sagittal,coronal,transverse TSEG T2 WIGFS and sagittal 3DGFLASHGWE sequences at 3.0T MR.The pathological changes of knee cartilage lesions,subarticular bone marrow edema (BME),bone cysts,bone attrition,osteophytes,meniscal inj ury,ligament inj ury,synovial thickening and joint effusion were evaluated according to the WORMS.The correlations between the above pathological changes and the pain were analyzed.Results There were significant and positive correlations between articular cartilage lesions and osteophytes,articular cartilage lesions and BME,BME and cyst,bone attrition and osteophytes,and BME and osteophytes,with the correlation coefficients of 0.6 70,0.5 9 8,0.5 1 8,0.5 1 4 and 0.505 ,respectively (P＜0.05 ).Multiple regression analysis showed that BME,bone attrition,synovial thickening and joint effusion were the main factors affecting the severity of pain after adj usted for age and BMI (P＜0.05).Conclusion The source of OA pain is multifactorial.The severity of BME,bone attrition,synovial thickening and joint effusion are the main factors associated with joint pain.As a nonGinvasive examination method,MR can make a comprehensive evaluation of the pathological changes of knee OA.

The 14-3-3 protein is a highly conserved acidic polypeptide family involved in intracellular signaling,protein transportation, cell proliferation, invasion, migration, and apoptosis. Many studies have shown that 14-3-3ζhas a high expression level in many malignant tumors. This paper introduces the structure of 14-3-3ζ,the biological function,and the development of the malignant tumor ( breast cancer,lung cancer,hepatocel-lular carcinoma,glioma,head and neck neoplasms) related to the development of 14-3-3ζand treatment of ma-lignant tumor in order to provide valuable information and ideas.

Objective To prepare PTD4-Cu,Zn-SOD fusion protein.Methods The recombinant plasmid of pET 1 6b-Cu,Zn-SOD and pET16b-PTD4-Cu,Zn-SOD was transformed into Escherichia coli BL21 (DE3).Isopropyl β-D-1-thiogalactopyranoside was then added at a final concentration of 0.84 mmol/L,and the cells were incubated for 4 h to induce the expression of Cu,Zn-SOD and PTD4-Cu,Zn-SOD fusion protein.Lysozyme and ultrasound were used to lyse the bacteria,the supernatant was collected for 15％ SDS-PAGE to analyze the expression of the target protein.Ni-NTA His bind resin was used to purify Cu,Zn-SOD protein and PTD4-Cu,Zn-SOD fusion protein under natural conditions.Western blot was used to identify the target protein.Results The results of Western blot showed that the purity of the target protein was about 90％,and the Cu,Zn-SOD protein with a molecular weight about 19 kDa and the PTD4-Cu,Zn-SOD fusion protein with a molecular weight about 20 kDa were found.Conclusion PTD4-Cu,Zn-SOD fusion protein is prepared successfully.

Objective: The study aimed to analyze the association between different types of obesity and dyslipidemia among rural primary and middle school students in Zhejiang Province, so as to inform strategies for prevention and control of childhood obesity and hyperlipidemia. Methods: As part of Nutrition Improvement Programme for Rural Compulsory Education Students, 1 244 participants were selected by stratified cluster random sampling in 5 counties of Zhejiang Province during September to December 2021. Physical examination, detection of blood lipid and questionnaire survey were conducted. The Chi -square test and Logistic regression analyses were used to assess the association between different types of obesity and dyslipidemia. Results: The prevalence rates of overweight, obesity, abdominal obesity, and hyperlipidemia were 15.11%, 12.46%, 17.60%, and 21.78%. Obesity and abdominal obesity were correlated to high risk of high triglycerides ( OR =3.97, 95% CI =2.54-6.20; OR =4.45, 95% CI =2.95- 6.72 )( P <0.05). Compared with the non overweight and obese group with normal waist circumference,the overweight and obesity group were correlated to high risk of high cholesterol ( OR=2.53, 95%CI =1.45-4.42, P <0.05). Abdominal overweight or obese group had the highest risk for dyslipidemia and triglycerides ( OR =1.82, 95% CI =1.33-2.48; OR =3.64, 95% CI =2.45-5.43) ( P < 0.05). Conclusions The prevalence rates of overweight, obesity, abdominal obesity, and hyperlipidemia are relatively high in rural primary and middle school students of Nutrition Improvement Programme for Rural Compulsory Education Students in Zhejiang Province. Abdominal obesity is a more important risk factor for hyperlipidemia. Waist circumference should be the focus of considerable attention.

Objective To explore the consistency of different physicians in diagnosis of malignant breast lesions with breast CEUS predictive model.Methods Totally 953 patients with solitary breast nodule from multicenter who underwent ultrasound and CEUS were collected.The research team was composed by the initial group (one junior physician from each hospital),check group (one or two physicians who had at least two-year experience of CEUS examination from each hospital),research group (two senior physicians from Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital) and cross-blinded group (one or two vice directors or chief physicians from each hospital).At first,the lesions were classified according to the breast imaging reporting and data system (BI RADS) by the initial group and the check group,then new BI RADS classifications were performed by research group and cross blind group with breast CEUS predictive model.The consistency of different physicians in diagnosis of malignant breast lesions was analyzed.Results Among 953 patients,benign lesions were found in 451 patients (451/953,47.32％),malignant lesions were found in 435 patients (435/953,45.65％),and precancerous lesions were found in 67 patients (67/953,7.03％).The accuracy of the initial group,check group,research group and cross-blinded group was 71.67％(683/953),74.92％(714/953),80.17％ (764/953) and 83.42 ％ (795/953),respectively.The consistency of different physicians for diagnosis of malignant breast lesions between initial group and check group was good (Kappa =0.82,P＜0.001),while between initial group and crossblinded group,initial group and research group were both moderate (Kappa =0.56,0.41;all P＜0.001).The consistency of different physicians for diagnosis of malignant breast lesions between cheek group and cross-blinded group,between check group and research group were both moderate (Kappa =0.68,0.51;all P＜0.001).The consistency between research group and cross-blinded group with breast CEUS predictive model was moderate (Kappa =0.74,P＜ 0.001).Conclusion The consistency of different physicians in diagnosis of malignant breast lesions with breast CEUS predictive model was moderate.

Objective: To investigate lunch supply of public primary school canteens in Zhejiang Province, and to provide a basis for the scientific guidance of school lunch. Methods: During May to Jun. and Sept. to Oct. in 2019, lunch food supply was weighed and recorded and the number of diners in 44 public primary school canteens were summarized. Each investigation lasted for one week. Results: Food was mainly based on the taste of the students (40.91%) in the school canteen. About 45.45% of the schools referred to the Nutrition Guidelines of School Meals for students meals when making recipes in the school canteen. The supplies of cereals, vegetables, fruits, livestock and poultry meat, fish and shrimp, eggs, milk, soybean nuts, vegetable oil and salt were 109.05, 118.01, 0, 63.96, 9.25, 11.31, 0, 10.68, 10.47, 2.54 g. The supply of vegetable oil was basically the same as the recommended amount ( P >0.05). The supplies of energy, protein, calcium, iron, zinc, vitamin A, vitamin B 1, vitamin B 2, vitamin C, dietary fiber were 820.84 kcal, 32.79 g, 164.18 mg, 7.84 mg, 4.71 mg, 23.07 μgRAE, 0.41 mg, 0.35 mg, 20.47 mg, 2.34 g, 37.56% of energy from fat and 48.47% of energy from carbohydrate. The supply of vitamin B 1 was basically the same as the recommended amount ( P >0.05). There were no significant differences in all kinds of food and nutrients between urban and rural primary schools ( P >0.05). Conclusion Lunch supply is not optimistic in public primary school canteens in Zhejiang Province, with unreasonable structure and fails to agree with current nutritional recommendations. It is suggested that the scientific guidance of students meals should be carried out according to the survey results combined with the characteristics of local diet.