1.Expression.purification and antigenicity of a recombinant respiratory syncyttal virus fusion gene encoded protein(amino acids 168-289)in the insect cells
Chinese Journal of Laboratory Medicine 2009;32(2):170-174
Objective To explore the antigenicity of the recombinant respiration syncytial virus (RSV)fusion protein (amino acids 168-289) encoded by 546-881 bases of the fusion gene expressed in insect baculoviruses expression system.Methods The fragment F' of fusion gene 546-881 bases was amplified from viral RNA( Long strain ) by the reverse transcription-polymerase chain reaction(RT-PCR).F' was inserted into transfer vector pBacPAK9 and the recombinant plasmid pBacRSV F' was constructed.Sf9 insect cells were then co-transfeeted with a mixture of recombinant plasmids pBacRSV F' and linearized BacPAK6 viral DNA( Bsu36 Ⅰ -digested).The recombinant baculoviruses BacPAK F' was constructed and was able to express the recombinant protein in Sf9 insect cells.The recombinant protein was purified by Ni2+ NTA chromatography and its antigenicity was identified by Western blot(WB) analysis.Specimens including the nasopharyngeal aspirates(NPAs) and sera were collected from 33 infants and young children with acute lower respiration tract infection. Indirect immunofluorecenee assay (IFA) and WB were used to detect the RSV antigen in NPAs and the anti-RSV antibody in sera respectively.Results The recombinant protein (molecular weight 13 000) was expressed in Sf9 insect cells.WB analysis demonstrated that the purified recombinant protein had a specific RSV antibody-binding activity. The recombinant protein could be recognized by positive serum infected by RSV.The positive rate was 27.3% and 33.3% respectively.There was no significant difference between them(X2 = 0.29 ,P > 0.05).Conclusion The recombinant respiratory syncytial virus fusion gene (546-881 bp) encoding protein is expressed in Sf9 insect cells and it has strong antigenicity and could have clinical application value for detection of RSV infection.
2.Application of the improved abdominal rotation card method in insulin injection
Wei YIN ; Shan FAN ; Zhijuan LI ; Hongmei GUO ; Hongbing BU
Chinese Journal of Practical Nursing 2016;32(22):1706-1709
Objective To explore the effect of the improved abdominal rotation card method in insulin injection. Methods A total of 100 hospitalized diabetes patients were randomly divided into control group (n=50) and observational group (n=50) according to the random number method. In the control group, insulin was injected to the subcutaneous tissue of abdomen with traditional method annular rotating method. Insulin was injected using improved abdominal rotation card method in the observational group. Compare accuracy and mastery rate of injection site rotation between the two groups. Compare fasting blood glucose (FBG), postprandial 2H blood glucose (PBG), HbA1c, the incidence of hypoglycemia and endermic induration between the two groups after three months. Results The nurses in the observation group had higher accuracy rate of the injection site rotation compared to the control group [98.6%(690/700) vs. 38.6%(270/700),χ2=584.66, P<0.01]. Mastery rate of the injection site rotation for the patients in the observation group were significantly higher than the control group [70.0% (35/50) vs. 20.0% (10/50), χ2=25.74, P < 0.01]. The incidence of endermic induration were significantly lower in observation group compared to the control group [2.0% (1/50) vs.16.0% (8/50), χ2=5.98, P < 0.01]. The incidence of hypoglycemia were significantly lower in observation group compared to the control group [4.0%(2/50) vs. 16.0%(8/50),χ2=4.00, P<0.01]. Conclusions The new abdominal rotation method in insulin injection can be a safe and effective therapy in patients with type 2 diabetes.
3.Improved effect of APP17 peptide on overexpression of phosphorylated Tau protein in brain tissues of mice with diabetes mellitus
Pengwen WANG ; Shan LU ; Yaping LEI ; Zhiwei ZHAO ; Zhijuan JI ; Shuli SHENG
Chinese Journal of Tissue Engineering Research 2006;10(44):202-203
BACKGROUND: Overexpression of phosphorylated Tau protein is a factor of dementia, and scholars abroad find that APP17 peptide may have effect on it.OBJECTIVE: To observe changes of phosphorylated Tau protein Ser202/Thr205 of mice with diabetes mellitus (DM) after injection of APP17 peptide.DESIGN: Randomized control study.SETTING: Department of Pathology, Capital University of Medical Sciences; Department of Brain Aging, Xuanwu Hospital, Capital University of Medical Sciences.MATERIALS: The experiment was carried out in the Pathological Department of Capital University of Medical Sciences and Brain Aging Department of Beijing Xuanwu Hospital. A total of 18 male Kunming mice of 8 weeks old and weighing 28-32 g were randomly divided into control group, DM group and APP17 peptide group with 6 in each group.METHODS: DM models were induced by streptozotocin (STZ) through selectively destroying β-islet cells; meanwhile, APP17 peptide was intraperitoneally injected into mice. Four weeks later, brain tissue underwentimmunohistochemical staining with AT-8 (Ser202/Thr205, a special monoclonal antibody).MAIN OUTCOME MEASURES: ① Morphological observation; ② AT-8 distribution; ③ quantitative analysis of immunohistochemical staining.RESULTS: Positive AT-8 cells in DM group were distributed in retrosplenial cortex, hippocampus, thalamus, hypothalamus, etc.; however, those incontrol and APP17 peptide groups were only distributed in retrosplenial cortex and hippocampus, and poorly stained.CONCLUSION: Positive AT-8 cells may be widely distributed in neurons of brains of DM mice; however, APP17 peptide may normalize the expression of positive AT-8 cells.
4.Investigation on undergraduates′cognition,attitude and behavior about low-carbon life
Shan FAN ; Qiuxia HAN ; Huamin LI ; Rugang JIANG ; Fang RUAN ; Zhijuan YU
Chongqing Medicine 2015;(18):2527-2530
Objective To investigate the undergraduates′cognition ,attitude and behavior about low‐carbon life and then to provide a relatively scientifically strategy .and provide scientific basis to formulate countermeasures about undergraduates′low‐car‐bon lifestyle .Methods By multistage sampling ,382 students′knowledge ,attitude and practice about low‐carbon life in Wuhan Uni‐versity of science and technology were analyzed .Results The students′cognition about low‐carbon life was good ,the awareness rate of girl was higher than that of boy ;Their attitude was positive ,grade and knowledge awareness were the two influence factors ;the behavior situation was unsatisfactory ,the origin of students and attitude enthusiasm were the elements affect rationality of be‐havior .Conclusion Undergraduates should strengthen the cognition of low‐carbon life ,improve attitude enthusiasm ,and form a good behavioral habit of low‐carbon life .
5.The effect of phytosterol intensive diet intervention in patients with type2 diabetes mellitus combined with nonalcoholic fatty liver
Wei YIN ; Ruo ZHUANG ; Qiaoyan LIU ; Shan FAN ; Zhijuan LI ; Hongbing BU ; Ruirong PAN
Chinese Journal of Practical Nursing 2017;33(28):2161-2167
Objectives To investigate the effects of phytosterol intensive diet intervention on blood glucose, blood lipid and liver function in patients with type 2 diabetes mellitus combined with nonalcoholic fatty liver disease(NAFLD). Methods Patients with NAFLD admitted to the department of endocrinology, the Affiliated Hospital of Jiangsu University from January 2016 to June 2016 were recruited.We divided the groups according to the order of patient admission,with patients admitted from January to March who received conventional diabetes mellitus low-fat diet enrolled as control group,and patients admitted from April to June received extra phytosterol intensive diet on the basis of conventional diabetes mellitus diet as treatment group. The changes of blood glucose, blood lipid and liver function between two groups with a follow-up of six months before and after intervention were compared and analyzed. Results After intervention,the levels of fasting blood sugar(FPG)and blood glucose(2hPG), glycosylated hemoglobin (HbA1c), cholesterol (TG), triglyceride (TC), alanine aminotransferase (ALT) of patients in control group(11.13 ± 3.17)mmol/L,(18.65 ± 6.21)mmol/L,(9.82 ± 1.69)%,(2.81 ± 1.43) mmol/L、(5.40 ± 1.14)mmol/L,77.27%(51/66),which were lower than those before intervention((8.51 ± 2.83)mmol/L,(10.39 ± 3.62)mmol/L,(7.78 ± 1.46)%,(2.18 ± 1.13)mmol/L,(4.99 ± 1.04)mmol/L, 90.91%(60/66),P<0.05,and FPG,2 hPG,HbA1c,TG,TC,LDL-C,ALT and aspartate aminotransferase (AST) in the experimental group were(11.32 ± 3.64)mmol/L,(20.09 ± 4.83)mmol/L,(9.70 ± 2.12)%, (2.68 ± 1.74)mmol/L,(5.16 ± 1.10)mmol/L,(3.18 ± 0.92)mmol/L,(70.27)%(52/74),(86.49)%(64/74), which were significantly lower than those before intervention((7.37 ± 2.08)mmol/L,(9.20 ± 3.35)mmol/L, (6.75 ± 0.99)%,(1.86 ± 1.13)mmol/L,(4.69 ± 1.06)mmol/L,(2.67 ± 0.72)mmol/L, 91.89%(68/74), 98.65%(73/74), P<0.05, and the differences was statistically significant(t=4.584,9.329,7.349,2.823, 2.140,χ2=4.587, P<0.01 or 0.05 in control group;t=8.106,15.715, 10.826,3.393,2.651,3.755,P<0.01 in experimental group). The levels of FPG, 2 hPG and HbA1c were significantly lower in the experimental group compared with those in control group after intervention(P<0.05),and the positive-to-negative rate of fatty liver were found to be significantly higher (33.8%,25/74) than that (9.1%,6/66) in controls(P<0.05).There were not significantly differences in the level of TG,TC,high density lipoprotein(HDL-C), LDL-C, ALT and AST between the control group and experimental group(P>0.05). Conclusions Phytosterol intensive diet intervention can effectively reduce LDL-C,AST and the blood glucose level of type 2 diabetes patients with NAFLD, improving the positive-to-negative rate of fatty liver. Phytosterol intensive diet intervention can effectively reduce LDL-C, AST and the blood glucose level of type 2 diabetes patients with NAFLD,improve the positive-to-negative rate of fatty liver.
6.Validity and reliability of the depression and somatic symptoms scale for screening depression among patients with coronary heart disease
Xinxin SHI ; Rongjing DING ; Zhijuan XIE ; Minjie ZHENG ; Shan QU
Chinese Journal of Behavioral Medicine and Brain Science 2022;31(12):1131-1136
Objective:To assess the validity and reliability of the depression and somatic symptoms scale among patients with coronary heart disease.Methods:Totally 246 patients with coronary heart disease were assessed with depression and somatic symptoms scale (DSSS), Hamilton depression rating scale for depression (HAMD) and patients’ health questionnaire depression scale-9 item (PHQ-9). The structural validity was evaluated with exploratory factor analysis and confirmatory factor analysis. The validity as a screening tool was evaluated with the gold standard diagnosed by psychiatrists who were trained with the mini international neuropsychological interview (MINI) according to ICD-10. Receiver operating characteristic (ROC) curve was used to identify cutoff scores for depression. Cronbach α coefficient was used to evaluate the internal consistency.Results:Exploratory factor analysis yielded two factors: depression factor and somatic factor, and the cumulative variance was 51.8%. The fitting indexes of confirmatory factor analysis were as follows: χ2/ df=3.636, RMR=0.077, RMSEA=0.104, IFI=0.804, TLI=0.781, CFI=0.802. The intraclass correlation coefficient of DSSS and HAMD was 0.54. The area under ROC curve (AUC) was 0.828, and the best boundary value was 17 points (sensitivity and specificity: 81% and 75%, respectively). The total scores and subscale scores for internal consistency of DSSS were higher in the depression group than those in the non-depression group ( P<0.01). Cronbach α coefficient for internal consistency of DSSS was 0.917. Conclusion:The DSSS has good validity and reliability among patients with coronary heart disease for screening depression, and can be used to screen depression among patients with coronary heart disease in general hospital.
7.High-performance liquid chromatography-mass spectrometry-based serum metabolic profiling in patients with HBV-related hepatocellular carcinoma.
Lei ZHANG ; Zhijuan FAN ; Hua KANG ; Yufan WANG ; Shuye LIU ; Zhongqiang SHAN
Journal of Southern Medical University 2019;39(1):49-56
OBJECTIVE:
To explore the diagnostic value of the serum metabolites identified by high-performance liquid chromatography-mass spectrometry (HPLC/MS) for hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC).
METHODS:
A total of 126 patients admitted to Tianjin Third Central Hospital were enrolled, including 27 patients with HBV-related hepatitis with negative viral DNA (DNA-N), 24 with HBV-related hepatitis with positive viral DNA, 24 with HBV-related liver cirrhosis, 27 with HBV-related HCC undergoing surgeries or radiofrequency ablation, and 24 with HBV-related HCC receiving interventional therapy, with 25 healthy volunteers as the normal control group. Serum samples were collected from all the subjects for HPLC/MS analysis, and the data were pretreated to establish an orthogonal partial least- squares discriminant analysis (OPLS-DA) model. The differential serum metabolites were preliminarily screened by comparisons between the HBV groups and the control group, and the characteristic metabolites were identified according to the results of non-parametric test. The potential clinical values of these characteristic metabolites were evaluated using receiver operator characteristic curve (ROC) analysis.
RESULTS:
A total of 25 characteristic metabolites were identified in the HBV- infected patients, including 9 lysophosphatidylcholines, 2 fatty acids, 17α-estradiol, sphinganine, 5-methylcytidine, vitamin K2, lysophosphatidic acid, glycocholic acid and 8 metabolites with few reports. The patients with HBV- related HCC showed 22 differential serum metabolites compared with the control group, 4 differential metabolites compared with patients with HBV-related liver cirrhosis; 10 differential metabolites were identified in patients with HBV-related HCC receiving interventional therapy compared with those receiving surgical resection or radiofrequency ablation. From the normal control group to HBV-related HCC treated by interventional therapy, many metabolites underwent variations following a similar pattern.
CONCLUSIONS
We identified 25 characteristic metabolites in patients with HBV-related HCC, and these metabolites may have potential clinical values in the diagnosis of HBV-related HCC. The continuous change of some of these metabolites may indicate the possibility of tumorigenesis, and some may also have indications for the choice of surgical approach.
Carcinoma, Hepatocellular
;
blood
;
diagnosis
;
virology
;
Case-Control Studies
;
Chromatography, High Pressure Liquid
;
DNA, Viral
;
blood
;
Hepatitis B virus
;
genetics
;
Hepatitis B, Chronic
;
blood
;
virology
;
Humans
;
Liver Cirrhosis
;
virology
;
Liver Neoplasms
;
blood
;
diagnosis
;
virology
;
Mass Spectrometry
;
Metabolome
;
Metabolomics
;
ROC Curve