1.Molecular cloning, prokaryotic expression and double-antibody sandwich ELISA development of 17β-hsd10 in mouse.
Chuanzhi LIU ; Yingying NIU ; Yuan'an CHEN ; Cheng WU ; Yuanhua YU
Chinese Journal of Biotechnology 2014;30(11):1774-1780
We expressed 17-hydroxysteroid dehydrogenase10 (17β-hsd10) recombinant protein, prepared anti-17β- hsd10 polyclonal antibodies and established sandwich enzyme linked immunosorbent assay (ELISA) test for detection of 17β-hsd10. RT-PCR was used to get the gene of 17β-hsd10 of mouse liver, and a prokaryotic protein expression system pET 15b-17β-hsd10/Escherichia coli BL21 (DE3) which induced with isopropyl-1-thio-β-galactopyranoside (IPTG) for recombinant protein expression was constructed subsequently. The target protein purified using His-Binding-resin column was used to immunize BALB/c mice and rabbits, serum total IgGs from immunized animals were purified by ammonium sulfate precipitation method. We established a Double-antibody Sandwich enzyme linked immunosorbent assay about 17β-hsd10 using the two antibodies we prepared. We got the concentration of 1.5 mg/mL of 17β-hsd10 protein with molecular weight of 29.5 kDa, and polyclonal antibodies from mouse and rabbit with the tite 1.25 x 10(4) and 2.5 x 10(4) respectively. The concentration of 0.1 g/mL of 17β-hsd10 can be detected by the Double-antibody Sandwich ELISA we established, and the assay was sensitive and specific. It can be widely used in clinical and experimental study.
3-Hydroxyacyl CoA Dehydrogenases
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genetics
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immunology
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Animals
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Antibodies
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immunology
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Cloning, Molecular
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Enzyme-Linked Immunosorbent Assay
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methods
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Escherichia coli
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Immunization
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Mice
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Mice, Inbred BALB C
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Rabbits
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Recombinant Proteins
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genetics
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immunology
2.Prevalence and rate of missed diagnosis study of COPD in presurgical patients
Yingying ZHOU ; Lijuan HU ; Li LIU ; Li LI ; Lei ZHU ; Shanfu NIU
Chinese Journal of Practical Internal Medicine 2000;0(12):-
Objective To invest the prevalence and the missed diagnosis rate of COPD in presurgical patients and their relationship with high risk factors.Methods Epidemic study was carried out in presurgical patients who received pulmonary function test in PFT laboratory of Zhongshan Hospital.Totally 339 patients were selected and their general conditions,exposure history and PFT results were analyzed.Results The prevalence of COPD was 21.8%,and the missed diagnosis rate was 83.8%.About 53.1% subjects had positive exposure history,among which 43.1% had smoking history and 18.6% had occupational exposure history,with the prevalence of 34.2% and 36.5%,between which no statistical significance was revealed.The prevalence of COPD in subjects with negative exposure history was 8.8%,and the missed diagnosis rate was 71.4%.The prevalence of COPD with and without exposure history above the age of 50 years was 38.0% and 10.0%,the former being significantly higher.The prevalence of COPD of men was 25.1%,which was significantly higher than that of women(15.2%)(P=0.04).Logistic regression analysis showed that the prevalence had little relation with gender(OR=0.80,95% CI:0.42~1.51),but had relation with exposure history(OR=4.22,95% CI:2.35~8.34).The severity classification of 62 COPD patients who were newly diagnosed after this PFT:stage Ⅰ 30.6%,stage Ⅱ 56.5%,stage Ⅲ 11.3%,stage Ⅳ 1.9%.Conclusion Most presurgical patients have the risk factors of COPD,like smoking and occupational exposure,and the latter is especially higher in our country.The prevalence and the rate of missed diagnosis of COPD is very high and the classification of severity of most missed diagnosed patients is stage Ⅱ or even more severe.
3.Inlfuence on Substance and Energy Metabolism of Diabetic Mouse Model of Chemical Split Fractions ofMori Cortex
Weisheng FENG ; Peipei YUAN ; Yan NIU ; Yingying KE ; Xiaolan WANG ; Zhiyi FENG ; Haixue KUANG ; Xiaoke ZHENG
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;17(3):464-470
This article was aimed to study the impact on substance and energy metabolism by chemical split fractions of Mori Cortex among hypoglycemic diabetic mouse model, in order to explain the new hypothesis of the science connotation in nature and flavor of traditional Chinese medicine (TCM). Male Kunming mice were intraperitoneally injected with a large dose of streptozotocin (STZ) (170 mg·kg-1) to establish type 1 diabetes mellitus mouse model. Medication was given consecutively for four weeks. The enzyme-linked immunesorbent assay (ELISA) was used to detect glucosekinase (GCK), glycogen phosphorylase (PYGL), pyruvate dehydrogenase (PDH), α-ketoglutarate dehydrogenase (α-KGDHC), phosphoglycerate kinase (PGK), acetyl coenzyme A (acetyl-CoA), adenylate kinase (ADK), fumarase (FUM), cytochrome C reductase (CCR), cytochrome C oxidase (COX) and other indicators. Enzymatic detection was used to determine the content of ATP coenzyme (ATPs), the content and ratio of NAD and NADH, the content of myocardial cell Na+-K+ ATP enzyme, as well as the content of ATP and ADP. The results showed that in the model group, the expression of PYGL was increased; and the expressions of GCK and PDH were decreased. It prompted that the source of glucose increased and the expelling of glucose decreased. The glucose level was increased. The COX expression was reduced and the respiratory chain was blocked. It regulated oxidative phosphorylation and the substrate phosphorylation level. It upregulated the expression of CCR, ATPs, NAD+, PGK, α-KGDHC and ADK. However, the expression of FUM was decreased. The activity of Na+-K+ ATPase was decreased significantly. At last, the metabolic disorders appeared. Mori Cortex aqueous extracts and the chemical split fractions significantly increased the GCK and PDH level in substance metabolism among diabetic mice. The levels of PYGL, α-KGDHC, PGK and acetyl-CoA were decreased (P < 0.05, or P <0.01). Meanwhile, it increased ATP and FUM, myocardial cell Na+-K+ ATP enzyme, and COX level in the energy metabolism (P < 0.05). It decreased the level of NAD+, CCR and ATPs (P < 0.05, or P <0.01). It was concluded that both the aqueous extracts and chemical split fractions of Mori Cortex can effectively improve the substance and energy metabolism disorders of diabetic mouse model. This effect may be related to the cold nature of Mori Cortex.
4.A comparative study for the effectiveness of the image by multiphase CT enterography among three scan phases
Jingjin LI ; Guoping XU ; Qinglai XIA ; Yingying LIU ; Xuening ZHANG ; Dongmei NIU ; Liangsheng LIU
Tianjin Medical Journal 2015;(9):1050-1053,1095
Objective To evaluate the influence of the CT enterography (CTE) in different scan phases of small intesti?nal wall and arteriovenous vessels, and to confirm the diagnostic efficiency thereof. Methods A total of 286 patients under?went CTE were collected, and their CT image of three scan phases, including 20 s, 40 s and 60 s after injection, were record?ed. Values of reinforcement of the intestinal wall and arteriovenous enhancing degree in all phases were collected. A statisti?cal analysis was used to confirm the best phase for showing the structure. The diagnostic efficiency of the best phase of CT image was evaluated for the clinical diagnosis of small intestine and vascular lesions. Results (1) The CT value of the small intestinal wall in the intestinal phase was significantly higher than other two phases (P<0.05). There was no significant dif?ference in CT value between arterial phase and portal phase. (2) Comparison between three phases of CT values of ileum showed that artery:arterial phase>intestinal phase, and there is no statistical difference between the intestinal phase and oth?er two phases. The result of CT value of ileum vein showed that intestinal phase>portal phase>arterial phase (P<0.05). (3) The sensitivity was 95.8%, the specificity was 94.9%, the accuracy was 95.1%, the positive predictive value was 79.3%and the negative predictive value was 99.1%using CT image of intestinal period for diagnosing small intestinal diseases. (4) The intestinal period showed small intestinal lesions and confirmed by pathology including 22 cases of small intestinal lympho?ma, 10 cases of stromal tumor, 1 case adenoma, cavernous 3 cases hemangioma and 8 cases of Crohn's disease. The diagno?sis of hemangioma should be combined with arterial phase. Conclusion Small intestinal wall and blood vessels have better display performance in the intestinal phase of multiphase CT enterography, using the arterial phase and intestinal phase scan?ning, we can make an accurate diagnosis for the majority of small intestinal diseases.
5.Impact of uric acid changing degrees on kidney function in middle-aged and elderly women——A follow-up cohort study
Min NIU ; Zhengnan GAO ; Shen LI ; Xue WEI ; Lan LUO ; Xiaolan GU ; Mingming LIU ; Nan ZHANG ; Ning LI ; Yingying DONG
Chinese Journal of Endocrinology and Metabolism 2017;33(5):392-397
Objective To determine the impact of uric acid changes on kidney function in middle-aged and elderly women with normal renal function.Methods Serum uric acid and estimated glomerular filtration rate(eGFR) were studied by prospective cohort analysis in 1152 middle-aged and elderly Dalian women with serum uric acid<7 mg/dl, estimated glomerular filtration rate(eGFR)≥90 ml·min-1·(1.73 m2)-1, and age>45 years old.Results The analysis of variance (ANOVA) of clinical indices between baseline and follow-up data showed that an eGFR was decreased after three years in both middle-aged and elderly women even with normal serum uric acid.Compared with those in the lowest guartile subgroup, a significant drop of eGFR to less than 90 ml·min-1·(1.73 m2)-1 was seen in those with the highest guartile of baseline serum uric acid group (OR=1.707, 95%CI 1.192-2.443, P=0.003).Furthermore, there were positive correlations between changes in the levels of serum uric acid and eGFR, and linear regression analysis showed that, as the change level of serum uric acid increased, the change level of eGFR declined(β=1.562, 95%CI 1.050-2.075, P<0.01], and the changing degree of uric acid increase is independent risk factors refor eGFR reductions.Conclusion Reduction in eGFR level is inversely related to serum uric acid in perimenopausal and postmenopausal women.However, our study showed that, women with serum uric acid level ranging from normal, slight increase, and relatively high levels, the increasing degrees of uric acid are independent risk factors responsible for the changing degree of eGFR reduction.
6.Experimental Study on Hypoglycemic Effect ofMori Cortex Aqueous Extracts and Chemical Split Fractions
Xiaoke ZHENG ; Peipei YUAN ; Yingying KE ; Shen WANG ; Aishe GAO ; Jinli HE ; Yangang CAO ; Na ZHANG ; Lingling LI ; Yan NIU ; Weisheng FENG
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(9):1957-1967
This article was aimed to study effects of aqueous extracts and all chemical split fractions ofMori Cortex on hypoglycemic effect of diabetic mice model. Intra-peritoneal injection of 170 mg·kg-1 streptozocin (STZ) was given to male Kunming mice to establish type I diabetes mode. Continuous administration of medication was given for 4 weeks. And then, indicators such as body weight, water intake, food intake, fasting plasma glucose (FBG), insulin, C-peptide, total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-c), and low density lipoprotein cholesterol (LDL-c) were detected. Pathological morphology of the liver and pancreas were observed by light microscopy. The results showed that high-dose group ofM. Cortex aqueous extracts can improve weight loss of type I diabetic mice, significantly reduce water intake and food intake, reduce FBG, TC, TG and LDL-c levels in different degrees (P<0.05 orP<0.01), and increase C-peptide and HDL-c levels (P<0.05 orP<0.01). When dosages of 30% ethanol fraction and fatty oil fraction were only about 1/2 and 1/4 ofM.Cortex aqueous extracts, we found that it can improve lipid disorder status,repair liver cells, and improve liver tissue damage. Its effect was superior to M. Cortex aqueous extracts. It was concluded thatM. Cortexaqueous extracts showed a better hypoglycemic effect. The effective component parts were 30% ethanol fraction and fatty oil fraction. Its hypoglycemic mechanism may be related to the promotion of insulin secretion, regulation of blood lipid disorders, as well as the protection of liver structure and function.
7.Progress in detection and technology applications of minimal residual disease detection in postoperative early stage non-small cell lung cancers
Ruiqi NIU ; Pingping LIU ; Yingying DU
Journal of International Oncology 2022;49(10):623-626
Surgery is the mainstay of treatment for early lung cancers, but there is still a risk of recurrence and metastasis after surgery. With the advancement of molecular biology and detection methods, detecting the level of postoperative peripheral blood minimal residual disease (MRD) in patients can dynamically monitor recurrence and determine prognosis. Due to the wide variety of MRD detection methods, uneven detection power, lack of uniform standards and prospective study validation, clinical application is still controversial. The further development of MRD detection for early stage cell lung cancer still needs technical progress, standardized detection criteria and credible clinical data.
8.Impact of prescribed adaptive statistical iterative reconstruction-V on image quality and radiation dosage of lung pure ground glass nodules: A phantom study
Ying LI ; Na'na AI ; Yingying ZHANG ; Yucun FU ; Jiong JIANG ; Dandan NIU ; Yanhui ZHAI ; Shouqiang JIA
Chinese Journal of Medical Imaging Technology 2018;34(5):775-778
Objective To investigate the impact of prescribed adaptive statistical iterative reconstruction V (Pre-ASiR-V) on imaging quality and radiation dosage of pure ground glass nodules (pGGN) in chest phantom,in order to obtain the optimal level of Pre-ASiR-V.Methods CT scanning for a chest phantom containing 4 artificial pGGNs was performed with Revolution CT,and the Pre-ASiR-V level was set as 0,20%,40%,60%,80% and 100% group,respectively.The mean noise,effective dose (ED) and the subjective scores of pGGN imaging were recorded and compared.Results The mean noise of groups (Pre-ASiR-V 0,20%,40%,60%,80% and 100%) was (17.93±2.20)HU,(17.30±3.68)HU,(18.20± 3.44)HU,(18.80±0.20)HU,(19.87±2.56)HU and (15.90±4.56)HU,respectively (F=0.568,P=0.723).ED of these groups was 7.40 mSv,5.16 mSv,3.36 mSv,1.97 mSv,0.97 mSv and 0.33 mSv,respectively.Compared with imaging of PreASiR-V 0,the reduction percentage of ED was 30.27%,54.59%,73.38%,86.89% and 95.54%,respectively.The subjective score of the image quality evaluated by the 2 observers had high agreement (Kappa=0.778,P=0.003),and all the scores were greater than 3.The subjective score of Pre-ASiR-V 80% and 100% group was slightly lower than those in other groups.Conclusion Different Pre-ASiR-V level slightly impacts the noise of high-resolution CT images of chest phantom,while reduces radiation dosage significantly.Pre-ASiR-V level of 60% is the optimal protocol.
9.An investigation of recognition and needs of policies of science and technology achievement transformation a-mong the medical personnel in 21 tertiary hospitals in Shanghai
Xiaojie HAN ; Yingying WANG ; Huiyao CHEN ; Xiaoxin YE ; Yuanyuan LI ; Xiaofeng XU ; Lu WEI ; Na LI ; Yun LIN ; Yuhong NIU ; Rong ZHOU ; Yang HONG
Chinese Journal of Medical Science Research Management 2019;32(4):277-280
Objective To understand the cognition ,attitudes and needs of the science and technology achievement trans-formation policies among the medical personnel of 21 tertiary hospitals in Shanghai .Methods Convenient sampling method was used .The self-designed questionnaires were used to survey 420 personals in 21 tertiary hospitals in Shanghai ,and 411 valid questionnaires were responded with a valid rate of 97 .86% .Results The cognition of science and technology achievement transformation policies of the respondents was low ,and the degree of cognition was related to the professional titles ( P<0 .05) .While the recognition degree of some policies was high ;40 .47% agreed that enterprises could be the decision-maker during the transformation process ;85 .99% agreed that it is OK to hire the Technology intermediary service agencies to evalu-ate the achievements value ,confirmed that need such agencies to provide the services of information collection and screening . Conclusions More Policy publicity should be enhanced among the medical personnel ,communications about their needs and suggestions on the science and technology achievement transformation should be carried out to further update related policies , as well as improve the transformation of science and technology achievements in health system .
10.Effect of neck CT arteriovenous simultaneous enhancement methods on image quality and radiation dose
Yunfu LIU ; Tianliang KANG ; Yongxian ZHANG ; Senlin GUO ; Lin FU ; Qinggang XU ; Yingying CAO ; Junfang XIAN ; Yantao NIU
Chinese Journal of Radiology 2023;57(7):756-761
Objective:To investigate the feasibility of simultaneous arteriovenous enhancement of neck CT with two-stage injection of contrast agent and its effect on image quality and radiation dose.Methods:A total of 30 patients undergoing neck CT enhancement scan due to space-occupying lesions in Beijing Tongren Hospital, Capital Medical University from February to April 2022 were prospectively included as the experimental group. The neck CT enhancement scan was performed with two-stage injection of contrast agent and arteriovenous simultaneous enhancement. The dosage of contrast agent was calculated according to the patient′s body weight, and the method of two-stage injection was adopted. The dosage of contrast agent in the first stage was 0.7 ml/kg, with normal saline in the middle stage, and the second stage (began at 35 s) was 0.3 ml/kg. A total of 30 patients with gender and age matching with the experimental group from December 2021 to January 2022 were retrospectively collected as the control group. The control group was treated with the traditional arterial phase and venous phase scanning method with the dosage of 1.0 ml/kg contrast agent. The arterial phase was scanned at the 30 s and the venous phase was scanned at the 60 s. The CT values of bilateral carotid arteries and jugular veins in the experimental group were measured, the CT values of bilateral carotid arteries in the arterial phase were measured in the control group, and the CT values of bilateral carotid arteries and jugular veins in the venous phase were measured. Carotid artery enhancement score was performed for images of experimental group and control group in arterial and venous phase, and jugular vein and lesion enhancement score was performed for images of experimental group and control group in venous phase. The effective dose was calculated for both groups. The difference of carotid artery CT values between images was compared by one-way analysis of variance, and LSD method was used for pairwise comparison. The CT values of jugular vein were compared using independent sample t test. Kruskal-Wallis test was used to compare carotid artery enhancement scores, and Nemenyi method was used for pairwise comparison. Jugular vein and lesion enhancement scores and effective dose were compared by Mann-Whitney U test. Results:The CT value of carotid artery of experimental group [left (276±24) HU, right (273±25) HU] was lower than that of control group in arterial phase [left (329±33) HU, right (327±32) HU], and higher than that in the venous phase [left (147±15) HU, right (148±16) HU]. All the differences were statistically significant ( P<0.001). The CT value of jugular vein of experimental group [left (206±18) HU, right (203±19)] was higher than that of control group in the venous phase [left (154±15) HU, right (151±15)], the difference was statistically significant ( t=11.88, 11.76, both P<0.001). There was no significant difference in carotid artery enhancement score between experimental group and control group in arterial phase ( P=0.624), but the carotid artery enhancement score of the experimental group was higher than that of the control group in the venous phase, and the difference was statistically significant ( P<0.001). The scores of jugular vein and lesion enhancement in experimental group were higher than those of control group in venous phase, and the difference was statistically significant ( Z=5.01, P<0.001). The effective dose of the experimental group [2.41(2.04, 2.72) mSv] was decreased by 52.2% compared with the control group [5.04(4.18, 5.44) mSv], and the difference was statistically significant ( Z=-6.24, P<0.001). Conclusions:The neck CT enhanced scan with two-stage injection of contrast agent and arteriovenous simultaneous enhancement method can obtain comprehensive images of arterial and venous phases, and realize simultaneous enhancement of carotid artery, jugular vein and lesions, and reduce radiation dose.