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 ; genetics ; immunology ; Animals ; Antibodies ; immunology ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; methods ; Escherichia coli ; Immunization ; Mice ; Mice, Inbred BALB C ; Rabbits ; Recombinant Proteins ; genetics ; immunology

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.

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.

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.

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.

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.

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.

Stroke is the world＇s second most dead disease and the leading cause of death in China. Its incidence, disability, recurrence rate, and high mortality rate, which seriously endangers the quality of life and life safety of residents. Continuing nursing can improve the health outcomes of stroke patients and reduce the rate of readmission. This article discusses the dynamic changes of continuing nursing needs of stroke patients from health education, life care, rehabilitation nursing, psychosocial support, etc., and analyzes the influencing factors of continuing nursing needs in terms of the general situation of patients, family society and health status, and provides a basis for future targeted hospital-community-family continuing nursing interventions.

Objective:To explore the influence of frequency of machinery heating power cleaning on dental handpiece loss and cleaning effect.Methods:This study selected 180 completely new high speed dental handpiece with machinery heating power cleaning. All of handpiece were randomly divided into group A, B and C, 60 cases in each group. Cleaning, oiling by special oiling machine, packing and sterilizing were carried out for the dental handpiece after being used. Group A, B and C implemented the process including machinery heating power cleaning, cleaning and oiling by special oiling machine, packing as well as sterilizing weekly, every other week and every three weeks respectively. This study randomly extracted 5 cases in each group to check the visible stains and monitor the cleaning effects after 3, 6 and 9 months.Results:Among group A, B and C, the using times were (56.60±24.77) , (49.70±30.08) , and (46.73±35.16) respectively with no statistical difference ( P=0.800) . There were 15 dental handpiece with trouble in 9 months, 4 in group A, 7 in group B and 4 in group C with no statistical difference ( P=0.660) ; 12 of them could be continued use after simple repair, 1 in group A, 7 in group B and 4 in group C with no statistical differences ( P=0.100) ; 3 of them could be continued use after changing central spindle, all in group A. The pass rates of visible cleanliness were all 100%. Conclusions:On the basis of cleaning and oiling by special oiling machine, machinery heating power cleaning every three weeks for dental handpiece can ensure the internal cleaning and improve working efficiency, but will not increase the loss.

Objective:To study changes of pollution conditions of the new dental unit waterlines.Methods:By means of random sampling, 7 new dental chair units of the same brand were selected. Of which, 5 dental chair units (DCU) were supplied with water after centralized treatment of municipal water and 2 dental chair units were supplied with independent water storage tanks. From the first day of clinical use, water samples of three-way syringes and high-speed dental handpieces and mouthwashes were continuously collected for bacterial culture and the number of colonies was counted.Results:A total of 343 water samples were collected. There was a statistically significant difference in the number of colonies in the water samples from the outlet of three-way syringes and high-speed dental handpieces after the waterlines was rinsed for 2 min before treatment ( P<0.01) . There was no statistically significant difference in the number of dental unit warterlines (DUWLs) colonies between the central water supply and the independent water storage tank before and after washing ( P> 0.05) . Waterline washing could significantly reduce the number of colonies in the waterlines ( P<0.01) , but there was no significant change in the number of colonies throughout the day ( P>0.05) . The number of colonies in the new DUWLs could exceed the recommended standard in the guideline on the second day of clinical use and the number of colonies increased significantly compared with that on the first day ( P<0.05) . Conclusions:In clinical work, no matter what kind of water supply method is used, in addition to waterline washing according to regulations, DUWLs should be disinfected daily.