Objective To investigate the distribution characteristics of motor endplates in musculuslevatorani of dif-ferent gestational ages. Methods 20~24 weeks female fetuses were categorized as Group A, and 24~28 weeks female fetuses as group B. Karnovsky-Roots iron cyanide copper staining ( hematoxylin) was used to study the aver-age weight, the morphology, distribution and changes in number of motor end plate. Results The motor endplates morphology and distribution of consistent oval of 20 ~28 weeks female fetuses showed a curve distribution in the musculuslevatorani fibers foreign 1/3 . The weight and numbers of motor endplates in Group A were less than those of group B. Conclusion 20 ~28 weeks female fetuses' musculuslevatorani are visible on the motor end plates, and no significant difference is found in morphology and distribution of the curve along the muscle fibers. The num-ber of motor endplates increases with the increase of anal muscle weight mentioned.

Objective To screen and identify the dengue virus-specific antigens,then establish the ELISA detection method for dengue virus antibody.Methods Using bioinformatic software DNAStar and ANTHEPROT to analyze the hydrophilicity,flexibility,surface probability and antigenicity of dengue virus type 1-4,Japanese encephalitis virus and Yellow fever virus M,E and NSI protein amino acid sequence and also consider the influence of secondary structure.Then in accordance with epitopes localion and amino acid sequence similarity,forecast the share and specific epitopes.Reference the sequence information of different dengue virus strains in GenBank to analyze the epitopes conservative.Based on the results of bioinformatic analysis,5 specific epitopes were amplified and inserted into prokaryotic expression vector pMAL-C2x or pET32a.Then the vectors was transferred into E.coli Rosetta( DE3 ).lsopropyl-β-D-thiogalactoside(IPTG) was used to induce the expression of gene segments.SDS-PAGE were used to identify the expression proteins,and the antigenicity were tested using Western blot.Using the antigen selected by Western blot,ELISA method for dengue virus antibody detection was established.Results Eighty shared epitopes and 25 specific epitopes were forecasted,and 5 antigenic fragments encloude analyzed epitopes from dengue virus type 2 and 3 were expressed in E.coli successfully.One dengue virus type 1-4 shared antigens (Den-Ag5),one dengue virus type 2 and 4 shared antigens( DenAg3),one dengue virus type 1-3 shared antigens(Den-Ag2) and two dengue virus type 1,2 and 4 shared antigens( Den-Ag1,Den-Ag4)were conformed using Western blot.Using antigens Den-Ag5,Den-Ag1 and DenAg2,the ELISA method for dengue virus antibody detection were established.Conclusion Based on the bioinformatic analysis and Western blot verification,5 dengue virus specific antigen were conformed,and the ELISA detection method for dengue virus antibody were established.

Objective To detect the serum prohibitin protein(PHB)level in children with renal interstitial damage and analyze the correlation between PHB and renal interstitial fibrosis(RIF). Methods Serum PHB protein levels were determined by Western blot analysis in 36 children with kidney diseases,and 30 healthy children were studied as control. Levels of BUN,Scr,and urinary microprotein series(including ALBU/Cr,NAGU/Cr,IgG U/Cr,α1-MU/Cr)were studied by automatic biochemical analyzer. Renal interstitial damage was semiquantitatively graded according to Katafuchi's method. The correlation between serum levels of serum PHB protein and those of BUN,Scr as well as urine microprotein were analyzed. Results Serum PHB protein was positive in children with diverse kidney diseases however it was negative in the normal controls(P ＜ 0.05). Serum PHB levels were significantly higher in children with proliferative glomerulonephritis than those with non-proliferative glomerulonephritis(P ＜ 0.05). Statistical analysis indicated that serum PHB levels positively correlated with the degree of tubulointerstitial lesions(r ＝ 0.868,P ＜ 0.001)as well as the glomerular injuries(r ＝ 0.753,P ＜ 0.001). And,serum PHB levels were also positively correlated with urinary microprotein including NAG(r ＝ 0.586,P ＜ 0.001)and IgG(r ＝ 0.341,P ＜ 0.001). Conclusions Serum PHB levels were significantly increased in children with kidney diseases and were positively correlated with the degrees of renal interstitial damage,suggesting that PHB might be a potential clinical marker for detecting tubulointerstitial lesions.

Objective To investigate the role of autophagy in radiation-induced death response of human nasopharyngeal carcinoma cells.Methods MTT method was used to detect cell viability of CNE-2 cells in different time after irradiation.Clonogenic survival assay was used to evaluate the effect of autophagy inhibitor (chloroquine phosphate) and autophagy inductor (rapamycin) on radiosensitivity of nasopharyngeal carcinoma cells.Cell apoptosis was assessed by flow cytometry.The expressions of LC3 and P62 were measured with Western blot.Cell ultrastructural analysis was performed under an electron microscope.Results Irradiation with 10 Gy induced a massive accumulation of autophagosomes accompanied with up-regulation of LC3-Ⅱ expression in CNE-2 cells.Compared with radiation alone,chloroquine phosphate (CDP) enhanced radiosensitivity significantly by decreasing cell viability (F =25.88,P ＜ 0.05),autophagic ratio (F =105.15,P ＜ 0.05),and LC3-Ⅱ protein level(F =231.68,P ＜0.05),while up-regulating the expression of P62 (F =117.52,P ＜ 0.05).Inhibition of autophagy increased radiation-induced apoptosis (F =143.72,P ＜ 0.05).Rapamycin (RAPA) also significantly decreased cell viability,but increased autophagic ratio and LC3-Ⅱ protein level while down-regulated the expression of P62.Induction of autophagy increased radiation-induced apoptosis(F =167.32,P ＜ 0.05).Conclusions Blockage of autophagy with CDP could enhance radiosensitivity in human nasopharyngeal carcinoma cells,suggesting that inhibition of autophagy could be used as an adjuvant treatment to nasopharyngeal carcinoma.

Objective To establish blood purification system for rats and provide a safe and reliable experimental platform for further research of blood purification. Methods The right carotid artery and the contralateral jugular vein of adult male Sprague?Dawley rats were cannulated to creat vascular access for blood purification, by which continuous arteriovenous hemofiltration blood purification system was established. Blood flow, substitution fluid flow and ultrafiltration rate were regulated by rotary mini?pumps. Blood purification therapy continued for 4 hours on the basis of maintained anesthesia and effective anticoagulation. The safety of continuous arteriovenous hemofiltration blood purification systems was evaluated by comparing the arterial blood gas, electrolyte indexes and blood glucose during the blood purification therapy. Closely monitoring the vital signs of rats, such as blood pressure and heart rate, and observing whether there were any side effects, such as massive haemorrhage, thrombogenesis and gas embolism in the therapeutic process. Results There were no obvious changes of arterial blood gas, electrolyte indexes and blood glucose during the blood purification therapy (P>0.05). The vital signs did not fluctuate acutely before and after the blood purification therapy (P>0.05). The incidence rate of side effects was very low. Conclusions
Continuous arteriovenous hemofiltration blood purification system had no obvious adverse effects on healthy rats. Our blood purification system for rats appears to be safe and reliable.

AIM:To observe the effect of the extract of Ginkgo biloba(EGB)on pituitary-testicular axis and the mRNA expressions of luteinizing hormone receptor(LHR)and steroidogenic acute regulatory protein(StAR).METHODS:Thirty male Sprague-Dauley rats were divided into three groups randomly:normal control group,type II diabetic group and EGB treatment group.After fed with high-fat diet for 4 weeks,the later two groups were injected with strepozotocin intraperitoneally to induce type II diabetes mellitus.The EGB treatment group was given EGB at the dose of 50 mg/kg once a day for 12 weeks by intragastric administration.The normal control and diabetic group were given normal saline of equal volume per day for 12 weeks.The indices of blood glucose,insulin and low-density lipoprotein-cholesterol(LDL-c)were measured.The morphologic change of testicular tissue was observed under light microscopy(LM)and transmission electron microscopy(TEM)respectively.The concentrations of blood luteinizing hormone(LH)and testosterone(T)were assayed by the technique of enzyme linked immunosorbent assay(ELISA).The mRNA expressions of LHR and StAR from Leydig cells were detected by RT-PCR.RESULTS:The concentrations of blood glucose,insulin and LDL-c increased obviously,and the testis weights lessened obviously in type II diabetic groups compared to those in normal control groups.Rare spermatogenic cells of seminiferous tubule and germinal arrest were observed in diabetic group under LM.Ultrastructural analysis of testicular tissue by TEM showed dilation of the endoplasmic reticulum and mitochondrial swelling in Leydig cell and sertoli cell in diabetic group.The level of blood LH and T decreased in typeⅡ diabetic groups in comparison with that in the normal control group.Compared to normal groups,the mRNA expression of StAR in type Ⅱ diabetic groups decreased,while the mRNA expression of LHR increased.After the treatment of EGB,the pathological change of testis was relieved,the concentrations of blood glucose,insulin and LDL-c were decreased,the level of blood LH and T,and the mRNA expression of StAR were increased,and the mRNA expression of LHR descended compared to type Ⅱ diabetic groups.CONCLUSION:EGB may increase the LH-induced testosterone production by correcting metabolic disorder of glucose and lipid,improving the function of pituitary-testicular axis and regulating the expression of LHR and StAR mRNA.

Objective: Construct a paraquat (PQ) cell fibrosis model in vitro, observe the effect of PQ on the expression of a disintegrin and metalloproteinase-17 (ADAM17) in A549 cells, and explore the role of ADAM17 in the pulmonary fibrosis induced by PQ poisoning. Methods: A549 cells are divided into normal control group, different concentration of PQ groups, CCK-8 is used to detect cell viability, screening concentration and time of PQ, cell morphology is observed under microscope; Enzyme-linked immunosorbent assay (ELISA) detectes fibrosis markers of collagen type I (Col I) and fibronectin (FN) expression. Establishment of cell model of fibrosis; distribution by immunocytochemical detection of ADAM17 in A549 cells, Reverse transcription-polymerase chain reaction and Western blot are used to detect the expression of ADAM17 mRNA and protein. Results: 1. With the increase of PQ concentration and the prolongation of the action time, the activity of A549 cells decreased (P< 0.05) , which is dose-dependent and time dependent. 2.The normal A549 cells fusion is paving stone growth and arranged more closely. After PQ induction, the cell arrangement was loose, the intercellular connection became loose, and some cells dissolved and died. 3.ELISA showed that with the increase of PQ concentration, the expression of Col I and FN increased (P<0.05) , and Col I and FN expression gradually increased with the prolongation of PQ time (P<0.05) , and the fibroblast model is successfully established. 4. Immunocytochemistry showes that ADAM17 is expressed in the cytoplasm of A549 cells. 5. RT-PCR and Western blot showed that the expression of ADAM17 mRNA and protein increased significantly with the increase of PQ concentration (P<0.05) , which is most obvious at PQ 200 μmol/L. With the prolonged action of PQ, the expression level of ADAM17 mRNA and protein also increased significantly (P<0.05) , and reached the peak in 24 h. Conclusion PQ can induce morphological changes of alveolar epithelial cells, cause cell damage, and successfully establish a cell fibrosis model, which has a dose and time dependence on the toxicity of A549 cells. ADAM17 is overexpressed in the A549 cells induced by PQ and may be involved in the process of pulmonary fibrosis induced by paraquat.

Objective:To research the application of deliberated Teach-back combined with Internet+nursing clinical pathway on Gartland type Ⅲ supracondylar fractures.Methods:A total of 51 children with Gartland type III supracondylar fractures who were hospitalized from March 2018 to May 2018 were selected as the control group, and 46 children with Gartlnd type III supracondylar fractures who were hospitalized from June 2018 to August 2018 were selected as the experimental group. The control group was given the traditional perioperative care routines, and the experimental group was given the Internet + perioperative clinical pathway combined with Teach-back for perioperative period.The execute accuracy of fasting and banned drinking was checked between the two groups, the incidence of postoperative needle infection was compared between the two groups, the function of the elbow joint was evaluated according to the Flynn score, the hospitalization days and hospitalization expenses were compared between the two groups.Results:The execute accuracy of fasting and banned drinking was 64.71%(33/51), 58.82%(30/51) in the control group and 86.96%(40/46), 84.78%(39/46)in the experimental group, and there were statistically significant differences between the two groups( χ2 values were 6.431, 7.937, P<0.05 or 0.01). The incidence of postoperative needle infection was 35.29%(18/51) in the control group and 2.17%(1/46) in the experimental group, and there were statistically significant differences between the two groups( χ2 value was 16.844, P<0.01). The difference of Flynn scores after 4 weeks operation between the two groups was statistically significant( P<0.01), but there was no significant difference after 6 weeks operation between the two groups( P>0.05). The hospitalization days and hospitalization expenses were(2.03±0.04) d, (10 135.403±94.384) yuan RMB in the experimental group, and (4.94±0.14) d, (12 577.646±192.604) yuan RMB in the control group, and there were statistically significant differences between the two groups( t values were 21.256, 11.386, P<0.01 or 0.05). Conclusions:The use of Teach-back combined with Internet + nursing clinical pathway on the Gartland type III supracondylar fracture of the humerus can improve the accuracy of preoperative fasting and banned drinking execut and reduce postoperative needle infection. The elbow function of the affected limb is significantly improved 4 weeks after operation. The hospitalization days and hospitalization expenses are reduced and the effect is significantly better than regular care, Which has the value of further clinical research and promotion and application.

Objective To compare the efficacy and safety of leflunomide (LEF) combined with medium/low dose corticosteroids and full dose of corticosteroids in the treatment of IgA nephropathy. Method Primary IgAN patients diagnosed by renal biopsy with 18?65 years old and eGFR≥30 ml·min?1·(1.73 m2)?1 and proteinuria>0.5 g/24 h were enrolled in a prospective controlled clinical study. They were randomly divided into leflunomide combined with medium/low dose corticosteroids (LEF group) and corticosteroids alone (steroid group). The primary outcomes were (1) end stage renal disease or dialysis (2) 50% increase in serum creatinine above the baseline. Secondary outcome was the remission of proteinuria. Results Ninety patients completed the follow?up. The 24?hour proteinuria at baseline were 2.00(1.10, 2.88) g and 1.87(1.13 ,3.08) g in LEF group and steroid group respectively. Compared with baseline, it was significantly decreased in both groups at 6 months [0.30(0.11, 0.93) g, 0.30(0.14, 1.33) g] and 12 months [0.30(0.09, 0.82) g, 0.32(0.14, 0.66) g], (P<0.05). Estimated glomerular filtration rate (eGFR) at baseline, 6 months and 12 months were (80.39 ± 28.56), (87.12±28.70) and (88.20±30.26) ml·min-1·(1.73 m2)-1. It was decreased in steroid group (P<0.05), while no significant difference was detected in LEF group[baseline (87.63 ± 27.35), 6 months (86.91 ± 32.45), 12 months (90.06 ± 30.00) ml·min-1·(1.73 m2)-1, P>0.05]. At 6 and 12 months, there was no significant difference in terms of 24?hour proteinuria, serum creatinine and eGFR (CKD?EPI) between groups (P>0.05). There was no statistically significant difference in adverse events between groups during the treatment (9/40 cases in LEF group and 11/50 cases in steroid group, P>0.05). The average follow?up was 79 months, and there was no difference in the renal prognosis between the two groups. Multivariate Cox regression analysis revealed that serum creatinine at baseline and renal interstitial inflammatory cell infiltration predicted the risk of the progress of IgA nephropathy. Conclusion Leflunomide plus medium/low dose corticosteroids has a similar effect as full dose of corticosteroids in IgA nephropathy and does not increase the risk for adverse events during the treatment.

Objective To compare FibroTouch, serum hepatic fibrosis index and aspartate aminotransferase-to-platelet ratio index (APRI) in diagnosis of liver fibrosis and early cirrhosis for patients with chronic HBV infection with mild abnormal liver function.Methods One hundred and seven chronic HBV infected patients with alanine transaminase (ALT)≤80 U／L who underwent liver biopsy in Mengchao Hepatobiliary Hospital of Fujian Medical University from January 2014 to January 2017 were enrolled in the study.According to the liver biopsy pathology , the patients were divided into mild liver fibrosis group (S1, n＝26 ), significant fibrosis group ( S2-3, n ＝34 ) and early cirrhosis group ( S4, n ＝47 ).The differentiations of liver stiffness measurement (LSM), Laminin(LN), Collagen Type ⅢN-peptide(PⅢP N-P), Collagen Type IV(CⅣ), Hyaluronic acid ( HA) and APRI were compared among the three groups , and their correlations with fibrosis were analyzed with Spearman rank correlation analysis .Logistic regression analysis was performed to establish a multi-index model for predicting the degree of liver fibrosis , and the receiver operating characteristic (ROC) curves were drawn to compare and analyze the predictive value for different stages of liver fibrosis.Results The LSM in the significant fibrosis group was higher than that in the mild liver fibrosis group (Z＝-2.611, P＜0.01).The levels of PⅢP N-P,CIV,LN and HA in early cirrhosis group were higher than those in mild liver fibrosis group (Z＝-4.026,-4.205,-2.938 and-3.156, all P＜0.01).LSM and APRI in early liver cirrhosis group was higher than that in mild liver fibrosis group(Z＝-5.789 and -4.308, both P＜0.01) and significant liver fibrosis group (Z＝-3.460 and -3.498, both P＜0.01).PLT in early liver cirrhosis group was lower than that in mild liver fibrosis group and significant liver fibrosis group (Z＝-4.533 and -3.206, both P＜0.01).The correlation analysis showed that the values of LSM , CIV, PⅢP N-P and APRI were positively correlated with the stage of liver fibrosis (r＝0.606, 0.418, 0.396 and 0.470, respectively; all P＜0.01).There was no significant difference in the efficacy of each single index in predicting hepatic fibrosis S≥2 and S＝4 (all P＞0.05). The prediction of liver fibrosis models were established with Logistic regression analysis .The model 1 Logit (P)＝-3.810+0.385×LSM+0.052×CIV was used for predicting liver fibrosis S≥2; the area under the receiver operating characteristic curve ( AUC) was 0.839, with specificity of 96.0%, and the diagnostic efficacy was superior to PⅢP N-P, CⅣand APRI(Z＝2.579, 2.145 and 2.219, all P＜0.05), but there was no significant difference compared with LSM ( Z ＝1.001, P ＞0.05).The model 2 Logit ( P)＝-4.291+0.105×LSM+0.028×CIV+2.682×APRI was used for predicting liver fibrosis S ＝4; its AUC was 0.865 with sensitivity of 82.9%and specificity of 81.8%, and the diagnostic efficacy was superior to LSM, PⅢP N-P, CⅣand APRI (Z＝2.309, 2.485, 2.403 and 2.103, all P＜0.05).Conclusions For chronic HBV infected patients with mild abnormal liver function , FibroTouch can be selected as a noninvasive method to predict liver fibrosis S≥2.The diagnostic efficacy of the prediction model 2, based on the combination of LSM, CIV and APRI is superior to each single index , which can improve the prediction level of early cirrhosis.