Objective To explore the mechanism of Exenatide-induced rat pancreatic tissue lesion.Methods Thirty SD male rats were divided into three groups according to complete random design,and each group had 10 rats,namely Exenatide group,diabetes-model group and control group.Diabetes-model rats were induced by streptozotocin (STZ,35mg/kg) and high-sugar and high-fat diet.The Exenatide group and diabetes group were subcutaneously administered with Exenatide at a dose of 5 μg/kg twice a day.The control group was treated with same amount of saline.Ten weeks later,all the rats were sacrificed and the pancreatic tissues were harvested for routine pathological examination.Immunohistochemical method was used to detect the expression of α-smooth muscle actin (α-SMA) and type Ⅲ collagen protein in pancreatic tissue,and ELISA was applied to measure the expression of matrix metalloprotei-nase-2 (MMP-2) and MMP-9 in pancreatic tissue.Results In control group,there was no pathological change in pancreatic tissue.In Exenatide group,chronic inflammatory changes were observed; and the degree of inflammatory changes were much severe in diabetes group,and the pathological scores were gradually increased in the 3 groups (P ＜0.05).The expressions of MMP 2 in pancreatic tissue in control group,Exenatide group,diabetes group were (186.98 ± 23.24),(306.07 ± 59.82),(365.08 ± 89.55) μg/L,and the expressions of MMP-9 were (49.37 ± 7.08),(67.24 ±14.73),(87.37 ±13.39)μg/L.The values were significantly higher in Exenatide group and diabetes group than those in control group (P ＜ 0.05),but the difference between the two groups was not statistically significant.The numbers of α-SMA positive cells per high power field were (13.4 ± 5.97),(29.5 ± 8.80),(79.3 ± 27.23) in control group,Exenatide group,diabetes group,and the numbers of type Ⅲ collagen positive cells were (10.6 ± 4.93),(29.3 ± 12.95),(56.0 ± 27.21).The values were significantly higher in Exenatide group than those in control group,and the values were significantly higher in diabetes group than those in Exenatide group (P ＜ 0.05).Conclusions Long-term subcutaneous injection of Exenatide may activate pancreatic stellate cells and cause expression of α-SMA,Ⅲ collagen protein,and MMP-2,MMP-9,then induce chronic inflammatory changes.

Objective To investigate the serum level of visfatin, C-reactive protein (CRP) and monocyte chemoattractant protein-1 (MCP-1) in patients with polycystic ovarian syndrome (PCOS), and provide the basis for understanding the pathogenesis of PCOS. Meth-ods 94 patients with PCOS and 100 healthy subjects were divided into 4 subgroups according to their body mass index(BMI), including obese PCOS subgroup(n = 52), non - obese PCOS subgroup (n = 42), obese healthy subject subgroup (n = 43) and non - obese healthy subject (n = 57). Serum visfatin, CRP, MCP-1, sex hormone levels and metabolic parameters were measured by enzyme-linked immu-nosorbent assay (ELISA), automatic chemistry analyzer or chemiluminescent immunoassay. Results Serum levels of testosterone (T), lu-teiizing hormone (LH) and prolactin (PRL) in the PCOS group were significantly higher than those in healthy subjects group(P <0.05～0.01), and follicle-stimulating hormone (FSH) levels were significantly decreased in PCOS group(P<0.01). Serum levels of Visfatin, CRP, MCP-1,fasting insulin(Fins) and insulin resistance homa model (HOMA-IR) in the obese or non - obese PCOS subgroup were signif-icantly increased than that in the obese or non - obese healthy subjects subgroup respectively (P<0.01). Pearson correlation analysis showed that the visfatin, CRP and MCP-1 levels were positively related to BMI, FINS and HOMA-IR(r=0.323～0.675, P<0.01). Par-tial correlation showed that serum visfatin levels were correlated with HOMA-IR(r=0.491, P<0.01), and serum MCP-1 levels were cor-related with LH (r=0.267, P<0.05) in the PCOS group. Conclusion The patients with PCOS had higher visfatin, CRP and MCP-1 lev-els, and visfatin levels were positively correlated with insulin resistance. Obesity were involved in the chronic inflammation course in patients with PCOS.

Objective To examine the expression and analyze the significance of autophagy-related gene microtubule-associated protein 1 light chain 3 (MAP1-LC3,LC3),p62 and lysosorne-associated membrane protein 2 (LAMP-2) in pancreatic tissues of mice with acute pancreatitis (AP).Methods Twenty mice were randomized into AP group and control group,and the number of mice was equal between two groups.AP group was intra-peritoneally injected by 20％ L-arginine solution (two injections of 4 g/kg body weight,every 1 h) in the dosage of 4 g/l kg twice every 1 hour to establish AP model,while control group was administered with equal volume of normal saline by intra-peritoneal injection.All the mice were euthanized at 24 hour after the last injection.Pancreatic histopathological changes were measured.In addition,the protein expressions of LC3,p62 and LAMP-2 were detected by Western blot.Results No obvious pathological changes were observed in control group.Pancreatic acinar edema,structure destruction,missing,the obvious widening of interlobular septum,small interlobular septum and acinar septum,and the necrosis of acinar cells at different degrees were observed in AP group.The pathological score for tissue edema,hemorrhage,necrosis and inflammation in AP group was 3.13 ± 0.50,2.83 ± 0.32,3.25 ± 0.46 and 3.16 ± 0.47,respectively,which was all 0 in control group.The differences were statistically significant between AP group and control group (P ＜ 0.01).In AP group,the ratio of LC3-Ⅱ/LC3-Ⅰ,p62 and LAMP-2 protein in pancreatic tissue were 1.16 ± 0.08,0.94 ± 0.04 and 0.35 ± 0.04,respectively,which were 0.24 ± 0.02,0.34 ± 0.03 and 0.95 ± 0.03 in control group.The ratio of LC3-Ⅱ/LC3-Ⅰ and p62 protein in pancreatic tissue in AP group were much higher than those in control group,while LAMP-2 in AP group was lower than that in control group,and there was statistically significant difference between two groups (all P ＜0.01).Conclusions Intraperitoneal injection of L-arginine could induce acute pancreatitis,and autophagy is impaired,which was associated with decreased LAMP-2 protein expression.

To analyze the structural characteristics of the Rap2B gene from Clonorchis sinensis by bioinformatics and to clone and express this gene through genetic engineering methods in order to obtain the corresponding recombinant protein.The structural and functional characteristics of the Rap2B protein were analyzed by using the related bioinformatic softwares. Using the full-length cDNA plasmid clone (Noc009e03) as template , the coding region of Rap2B gene sequence was amplified with PCR and cloned into the prokaryotic expression vector pET-28a(+) and then expressed in E.coli BL21 through IPIG induction. The recombinant protein expressed was purified by Ni-IDA affinity chromatography and detected by SDS-PAGE and Western blotting. It was demonstrated that the size of this gene was 847 bp in length, encoding 141 amino acids and its theoretical molecular weight was 15851.1 daltons. As demonstrated by PCR, double enzyme digestion and DNA sequencing, the recombinant expression plasmid pET-28a(+)-Rap2B was successively constructed and the Rap2B-like gene was highly expressed in E.coli BL21. The recombinant protein was obtained through purification with His affinity chromatography and could react specifically with the serum of rats infected with C.sinensis. Through these investigations, the Rap2B protein may be predicted as a member of the Ras oncogene family protein and is potential in the carcinogenic process of C.sinensis.

Objective To explore the alteration and effect of autophagy in pancreas tissue of rat injected by exenatide.Methods Diabetes model rats were induced by two-month high-sugar and high-fat diet and streptozotocin injection (35 mg/kg) in normal rats.50 SD male rats were divided into four groups according to the principle of complete random design,namely normal control group (n=10),normal exenatide-injected group (n=10),diabetes-model control group (n=l5) and diabetes-model exenatide-injected group (n=15).Rats in exenatide-injected groups were subcutaneously injected with exenatide respectively in 5 μg/kg dose each time,twice a day,at 8 a.m.and 6 p.m.Animal weights were weighted weekly and the dose of exenatide was adjusted according to current weight.Rats in the two control groups were injected with the corresponding amount of saline.Mter 10 weeks of treatment,all rats were killed and pancreatic tissues were disposed.Immunohistochemistry was used to measure the expression of GLP-1R in pancreatic tissues.Western blot was used to test the expressions of LC3-Ⅰ,LC3-Ⅱ and p62 in pancreatic tissues,and LC3-Ⅱ/Ⅰ ratio and p62 were compared between any two groups.All specimens were stained with hematoxylin-eosin (HE).The data were expressed as means ± standard deviation and were analyzed by unpaired Student t test using SPSS 18.0 statistics software.P value ＜0.05 was considered to be statistically significant for all tests.Results The pancreatic tissues from 13 rats (6 from the normal exenatide-injected group and 7 from the diabetes-model exenatide-injected group) appeared pathological changes such as gland structure damage,pancreatic cells atrophy and cells compartment broadening.The expressions of GLP-1R,LC3-Ⅱ and p62,and LC3B-Ⅱ/Ⅰ ratio in the two exenatide-injected groups were higher than those in the respective control group,and the differences had statistical significance (P＜0.05).Conclusions Long-term subcutaneous injection of exenatide can upregulate the expression of GLP-1R in rat pancreatic acinar cells and may induce the impaiment of autophagy flux in rat pancreatic cell.

Objective To study the correlations of VDR FokⅠgene polymorphism with type 2 diabetes in postmenopausal women of Han nationality in south Sichuan. Methods 160 patients with type 2 diabetes (T2DM) and 190 healthy cases were enrolled in the study. The VDR FokⅠgene polymorphisms were detected using RFLP-PCR and DNA sequencing. Results The FF, Ff and ff genotype frequencies were 32.5%, 47.5%and 20% in the T2DM group and 15.8%, 53.7%, 30.5% in the control group, respectively (P < 0.05). The allele frequencies were 56.3%, 43.8% in the T2DM group and 42.6%, 57.4% in the control group, respectively (P < 0.05). The risk of T2DM in the FF genotype people was 2.568 times higher than Ff/ff genotype (adjusted OR = 2.568, 95%CI = 1.246 ~ 5.292, P < 0.05). The levels of 2 h PG and HbA1C in the FF genotype people were significantly higher than those of the Ff/ff genotype people (P<0.05). Conclusions There was an association between the VDR FokⅠgene polymorphism and type 2 diabetes incidence in the postmenopausal women in south Sichuan area.

Background:Surgical treatment of both-column acetabular fractures is challenging because of the complex acetabular fracture patterns and the curved surface of the acetabulum.Seldom study has compared the application of three-dimensional (3D) printing technology and traditional methods of contouring plates intra-operatively for the surgical treatment of both-column acetabular fractures.We presented the use of both 3D printing technology and a virtual simulation in pre-operative planning for both-column acetabular fractures.We hypothesized that 3D printing technology will assist orthopedic surgeons in shortening the surgical time and improving the clinical outcomes.Methods:Forty patients with both-column acetabular fractures were recruited in the randomized prospective case-control study from September 2013 to September 2017 for this prospective study (No.ChiCTR1900028230).We allocated the patients to two groups using block randomization (3D printing group,n =20;conventional method group,n =20).For the 3D printing group,1:1 scaled pelvic models were created using 3D printing,and the plates were pre-contoured according to the pelvic models.The plates for the conventional method group were contoured during the operation without 3D printed pelvic models.The operation time,instrumentation time,time of intra-operative fluoroscopy,blood loss,number of times the approach was performed,blood transfusion,post-operative fracture reduction quality,hip joint function,and complications were recorded and compared between the two groups.Results:The operation and instrumentation times in the 3D printing group were significantly shorter (130.8 ± 29.2 min,t =-7.5,P ＜ 0.001 and 32.1 ± 9.5 min,t =-6.5,P ＜ 0.001,respectively) than those in the conventional method group.The amount of blood loss and blood transfusion in the 3D printing group were significandy lower (500 [400,800] mL,Mann-Whitney U=74.5,P ＜ 0.001 and 0 [0,400] mL,Mann-Whitney U =59.5,P ＜ 0.001,respectively) than those in the conventional method group.The number of the approach performed in the 3D printing group was significantly smaller than that in the conventional method group (pararectus + Kocher-Langenbeck [K-L] approach rate:35％ vs.85％;X2 =10.4,P ＜ 0.05).The time of intra-operative fluoroscopy in the 3D printing group was significantly shorter than that in the conventional method group (4.2 ± 1.8 vs.7.7 ± 2.6 s;t =-5.0,P ＜ 0.001).The post-operative fracture reduction quality in the 3D printing group was significantly better than that in the conventional method group (good reduction rate:80％ vs.30％;X2 =10.1,P ＜ 0.05).The hip joint function (based on the Harris score 1 year after the operation) in the 3D printing group was significantly better than that in the conventional method group (excellengood rate:75％ vs.30％;x2 =8.1,P ＜ 0.05).The complication was similar in both groups (5.0 ％ vs.25 ％;x2=3.1,P =0.182).Conclusions:The use of a pre-operative virtual simulation and 3D printing technology is a more effective method for treating bothcolumn acetabular fractures.This method can shorten the operation and instrumentation times,reduce Mood loss,blood transfusion and the time of intra-operative fluoroscopy,and improve the post-operative fracture reduction quality.

Objective To investigate the influences of verapanal preconditioning on cardiac function in vitro and intracellular free Ca~(2+) and L-type calcium current (I_(Ca-L)) in rat cardiomyocytes post ischemiareperfusion (I/R) injury. Methods The isolated rat hearts in control group (37 ℃ Tyrode solution perfusion for 30 min, n =6), I/R group ( no flow for 30 min followed 30 min reperfusion with 37 ℃ Tyrode solution, n =7) and verapamil preconditioning group [37 ℃ Tyrode solution perfusion for 10 min, adding verapamil (20 μmol/L ) to Tyrade solution and perfusion for another 30 nan, followed then by 30 min no flow and 30 min reperfusion, n=7 ] using Langendorff perfusion system. The fluorescence intensity of intracellular Ca~(2+) was detected with Fluo-3/AM loading by the laser scanning confocal microscope. The I_(Ca-L) was recorded via whole-cell patch clamp technique in enzymatically dissociated single rat ventricular myocytes. Results As expected, arrhythmias and cardiac dysfunction were shown post I/R injury. The fluorescence intensities of intracellular free Ca~(2+) in cardiomyocytes were significantly increased compared with control group (P <0. 01 ). By voltage clamp protocol, peak current densities of I_(Ca-L) was significantly reduced and I-V curve significantly elevated. Post I/R injury compared with control group (P < 0. 01 )which could be reversed by Verapamil preconditioning. Verapamil preconditioning also significantly improved diastolic and systolic functions, and reduced the incidence of arrhythmias. Conclusions Myocardial I/R injury might significantly impair heart functions and induce arrhythmias via cellular Ca~(2+) overload.Verapamil preconditioning could prevent heart I/R injury and reduce arrhythmias by decreasing influx of I_(Ca-L), thereby stabilizing cardiomyocytes in myocardial stunning and avoiding occurrence of Ca~(2+) -induced Ca~(2+) release during I/R injury.

AIM:To elucidate the association between chronic kidney injury and interleukin-33 (IL-33;an alarmin)/suppression of tumorigencity 2 (ST2) in patients with systemic lupus erythematosus (SLE).METHODS:Serum levels of IL-33 and soluble ST2 (sST2) were assessed by ELISA in 50 SLE patients and 30 healthy controls (HC).RESULTS:The levels of IL-33 and sST2,and IL-33/sST2 ratio were significantly higher in SLE patients than those in the HC.The IL-33 and sST2 levels were positively associated with SLE disease activity index (SLEDAI),erythrocyte sedimentation rate (ESR),proteinuria and triglyceride,but negatively associated with complement C3.IL-33/sST2 ratio was positively associated with SLEDAI and estimated glomerular filtration rate (eGFR).Independent explanatory variables associated with high IL-33/sST2 included chronic kidney disease (CKD) staging and albumin (R2 =0.442),especially CKD staging.CONCLUSION:Elevated serum sST2 and IL-33 levels in SLE patients are correlated with disease activity and risk factors of kidney injury.IL-33/sST2 ratio may serve as a potential biomarker for chronic kidney injury in SLE patients.

Three kinds of glass sharp electrospray ionization devices were built for direct mass spectrometry (MS) analysis using sharp slide,slender capillary and sharp dropper. Through the simulation experiment and mass spectrometry experimental investigation,the influence of the experimental conditions was studied. Under the optimum working conditions such as the electrospray tip diameter was less than 0.24 mm, the distance between the electrospray tip and the MS sample inlet was 10 mm, the electrospray voltage was 4.5 kV, the electrospray capillary angle was about 5.0°-7.5° (upper at back) and the left and right angles were 0°, 0.1 mg/mL sample solution(containing 1% formic acid for positive ion detection,or ammonium hydroxide for negative ion detection) could be well analyzed by MS. In positive or negative ion modes, the quinine, mandelic acid,benzoin,phenylalanine and other different types of sample solutions were analyzed by MS. The result showed that quinine,mandelic acid,benzoin,phenylalanine,tartaric acid,2,5-dihydroxybenzoic acid (DHB) and tangeretin could be detected in positive ion mode using sharp dropper electrospray. DHB, phenylalanine, tartaric acid and mandelic acid were detected in negative ion mode. Sharp slide electrospray was suitable for measuring the stable sample in the air, capillary electrospray and sharp dropper electrospray were suitable for detecting many low viscosity sample solution even instable sample. Using the method, the extraction solution of propranolol hydrochloride tablet and berberine hydrochloride tablet was directly analyzed by MS. The results showed that the glass sharp electrospray method had the advantages such as without carrier gas, simple structure, easy to build and operate, rapid analysis, anti-cross pollution, and could be conveniently used for MS analysis.