AlM: To study the effects of different concentrations of Coix seed oil on human retinal capillary endothelial cells ( HRCECs ) proliferation and vascular endothelial growth factor ( VEGF) expression in high glucose environment.
METHODS: HRCECs extracted from human fresher eyeball and cultured in vitro, and ultimately used in the experiment were the growth of 3rd ~ 4th cells, the experimental were divided into blank control group, low glucose control group, high glucose control group, high glucose + ( 50ü L/mL, 100ü L/mL, 200ü L/mL ) different concentrations Coix seed oil group. Detecting the multiplication of HRCECs by MTT, the immunocytochemical method was employed to detect the each group HRCECs of VEGF expression.
RESULTS:MTT assay results showed that: different concentrations of coix seed oil acted at HRCECs for 48h, inhibition of cell proliferation was significant difference compared with high glucose control group (P<0. 05). Within 48h showed concentration dependence. There was no statistical difference between the low glucose group and high glucose control group (P>0. 05). lmmunocytochemical assay showed that:50ü L/mL, 100ü L/mL, 200ü L/mL Coix seed oil acted at HRCECs 48h, the expression of VEGF decreased significantly compared with the high glucose control group ( P< 0. 05 ), and in a dose- dependent manner. However, in high glucose control group, the expression of VEGF was obvious higher than that of low glucose control group (P<0. 05).
CONCLUSlON:Coix seed oil can inhibit the HRCECs proliferation and suppress the VEGF expression in high glucose environment.

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.

AIM: To examine the relationship between corneal lymphangiogenesis and hemangiogenesis after keratoplasty.METHODS: Nineteen human corneas were obtained from 19 patients undergoing a second corneal transplantation in Zhongshan Ophthalmic Center in 2005. Blood and lymphatic vessels in human transplanted corneas were identified by lymphatic vessel endothelial receptor(LYVE-1) and platelet endothelial cell adhesion modecule-1(PECAM-1) immunohistochemistry, and double enzyme-histochemistry; then the association of corneal blood vessel counting (BVC) with lymphatic vessel counting (LVC) was examined.RESULTS: Corneal hemangiogenesis was present in 12 cases (63%), and lymphangiogenesis occurred in 5 cases (26%) of human transplanted corneas. In addition, corneal lymphangiogenesis was only present in vascularized corneas. LVC was strongly and positively correlated with BVC(r=0.725;P<0.01).CONCLUSION: Corneal lymphangiogenesis develops after keratoplasty, and strongly associates with hemangiogenesis.

Objective The purpose of this study was to develop a high-throughput micro-plate radiobinding assay (RBA) of glutamic acid decarboxylase antibody (GAD-Ab) and to evaluate its clinical application. Methods 35labeled GAD65 antigen was incubated with sera for 24 h on a 96-well plate, and then transferred to the Millipore plate coated with protein A, which was washed with 4℃ PBS buffer, and then counted by a liquid scintillation counter. The GAD-Ab results were expressed by WHO standard unit (U/ml). A total of 224 healthy controls, 162 patients with type 1 diabetes mellitus(T1DM) and 210 patients with newly diagnosed type 2 diabetes (T2DM) were recruited. A total of 119 TI DM and healthy cases with gradually changing GAD-Ab levels were selected to compare the consistency of micro-plate RBA with conventional radioligand assay (RLA). Blood samples were obtained from the peripheral vein and finger tip in 32 healthy controls, 35 T1DM and 24 T2DM patients, and tested with micro-plate RBA and then compared with the conventional RLA to investigate the reliability of finger tip sampling. Linear correlation,student's t-test, variance analysis and receiver operating characteristic (ROC) curve were performed using SPSS 11.5. Results (1) The optimized conditions of micro-plate RBA included 2 μl serum incubated with3 ×104 counts/min 35S-GAD for 24 h under slow vibration, antigen-antibody compounds washed 10 times by 4℃ PBS buffer, and radioactivity counted with Optiphase Supermix scintillation liquid. (2)The intra-batch CV of the micro-plate RBA was 3.8%- 10.2%, and the inter-batch CV was 5.6%- 11.9%. The linearity analysis showed a good correlation when the GAD-Ab in serum samples ranged from 40.3 to 664 U/ml and the detection limit of measurement was 3.6 U/ml. The results from Diabetes Autoantibody Standardization Program (DASP) 2005 showed that the sensitivity and specificity for GAD-Ab were 78% (39 positive among 50 new-onset T1DM) and 98% (2 positive among 100 healthy controls). The results of GAD-Ab obtained with micro-plate RBA and RLA were closely correlated (r=0.915,P<0.001) with a high concordance level of 97.5% and a Kappa value of 0.95. (3)TI DM and T2DM patients showed higher positive rates for GAD-Ab than the healthy controls(46.9% and 5.2% vs 0.89% ,X2=123.5 and 10. 1 ,P <0.001 and <0.01, respectively). (4)The consistency of GAD-Ab measurement with RBA using finger tip blood and RLA measurement using venous blood was 96.7% (r =0.946,P <0.001, Kappa value: 0.905). Conclusions The micro-plate RBA of GAD-Ab has high sensitivity, specificity and reproducibility, and can be measured with finger tip blood sampling. It might be a better alternative for clinical practice.

Objective To evaluate the value of 18F-FDG PET/CT in preoperative diagnosis and staging of suspected extrahepatic cholangiocarcinoma (EHCC).Methods The clinical data of 116 patients (72 males,44 females;age range 26-89 years) with suspected EHCC from January 2013 to October 2014 were retrospectively analyzed.All patients received preoperative whole body 18F-FDG PET/CT scan.The imaging results were compared with final clinical diagnosis.The diagnostic sensitivity,specificity,positive predictive value,negative predictive value and accuracy of 18F-FDG PET/CT were calculated.Two-sample t test was applied to compare lesion SUVmax of malignant and benign diseases.One-way analysis of variance was applied to compare SUVmax of highly,moderately and poorly differentiated tumors.x2 test was used to compare the difference of diagnostic sensitivities for hilar cholangiocarcinomas and common bile duct tumors.Results All patients were confirmed by exploratory laparotomy and subsequent histologic examination.A total of 94 cases (93 adenocarcinomas and 1 squamous carcinoma) were confirmed malignant and 22 cases (11 biliary calculi,9 cholangitis,1 choledochal cyst,1 tuberculosis) were confirmed benign.The diagnostic sensitivity,specificity,positive predictive value,negative predictive value and accuracy of 18F-FDG PET/CT for primary tumor were 61.7％ (58/94),77.3％ (17/22),92.1％ (58/63),32.1％ (17/53),64.7％ (75/116),respectively.The diagnostic sensitivity and specificity for regional lymph node metastasis were 45.5％(15/33),91.4％(53/58),and those for distant metastasis were 3/4,94.3％(82/87).The SUVmax of malignant tumors were higher than that of benign lesions (4.57± 3.75,2.72± 2.48;t =2.83,P＜ 0.05),while the differences of SUVmax among highly,moderately and poorly differentiated tumors were not significant (4.89±4.75,4.23±2.49,4.47±2.73;F=0.269,P＞0.05).18F-FDG PET/CT showed a lower sensitivity in hilar cholangiocarcinomas than that in common bile duct tumors,while no statistical significance was observed:48.6％ (17/35) vs 69.0％ (40/58),x2=3.827,P＞0.05.Conclusions The value of 18F-FDG PET/CT in preoperative diagnosis and staging of EHCC is limited.It can distinguish some benign diseases from malignant tumors,but with higher false positive for cholangitis.It can help to adjust treatment strategies by detecting distant metastasis.

OBJECTIVE: To investigate the risk factors for hypoglycemia after birth in preterm infants with a gestational age of ≤32 weeks. METHODS: A retrospective analysis was performed for 86 neonates with hypoglycemia and a gestational age of ≤32 weeks who were admitted to the neonatal intensive care unit from January 2017 to June 2020 (hypoglycemia group). A total of 172 preterm infants with normal blood glucose who were hospitalized during the same period were randomly enrolled as the control group. Univariate analysis and multivariate logistic regression analysis were used to screen out the risk factors for hypoglycemia in preterm infants. RESULTS: There were 515 preterm infants during the study, among whom 86 (16.7%) had hypoglycemia. Compared with the control group, the hypoglycemia group had significantly higher percentages of small for gestational age (SGA), cesarean section, maternal hypertension, and antenatal steroid administration (P<0.05), but significantly lower birth weight and rate of intravenous glucose use before blood glucose test (P<0.05). SGA (OR=4.311, 95%CI: 1.285-14.462, P<0.05), maternal hypertension (OR=2.469, 95%CI: 1.310-4.652, P<0.05), and antenatal steroid administration (OR=6.337, 95%CI: 1.430-28.095, P<0.05) were risk factors for hypoglycemia in preterm infants, while intravenous glucose use (OR=0.318, 95%CI: 0.171-0.591, P<0.05) was a protective factor against hypoglycemia in preterm infants. CONCLUSIONS SGA, maternal hypertension, and antenatal steroid administration may increase the risk of early hypoglycemia in preterm infants with a gestational age of ≤32 weeks, and intravenous glucose use is recommended as soon as possible after birth for preterm infants with a gestational age of ≤32 weeks to reduce the incidence rate of hypoglycemia.
Cesarean Section ; Female ; Gestational Age ; Humans ; Hypoglycemia/etiology* ; Infant ; Infant, Newborn ; Infant, Premature ; Infant, Small for Gestational Age ; Pregnancy ; Retrospective Studies ; Risk Factors

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.

Humans ; Liver Function Tests ; methods ; Technetium Tc 99m Aggregated Albumin ; Technetium Tc 99m Pentetate

Objective To evaluate the CT findings of pancreatic carcinoid tumors.Methods The CT imaging data of five patients with pancreatic carcinoid tumors confirmed by pathology were retrospectively analyzed.Results The tumors ranged in maximum diameter from 2.0 to 11.0 cm with a mean of 6.4 cm. On unenhanced CT,the tumors were slightly hypodense relative to the pancreatic parenchyma,homogenous in 2 cases,and heterogenous in 3 cases.One tumor showed calcification.After contrast material injection, the solid component of the tumor showed marked heterogenous enhancement on the arterial phase scanning in 3 cases,and mild heterogenous enhancement in 2 cases.The degree of tumor enhancement was less intense than the surrounding pancreatic parenchyma due to necrosis of various degree,which led to the cystic appearance of the tumor in 1 ease.On the portal phase scanning,all tumors showed marked enhancement similar to that of the pancreatic parenchyma.On the delayed phase scanning,the degree of enhancement was more intense than the surrounding pancreatic parenchyma in 1 case.Liver metastases with retroperitoneal lymphadenopathy and peripancreatic vessels invasion were seen in 1 case.No dilatation of the biliary tract or pancreatic duct was present.Conclusion The CT features of pancreatic carcinoid tumors included infrequent dilatation of the biliary tract or pancreatic duct and unusual vascular involvement,calcification within the mass,marked enhancement similar to that of the surrounding pancreatic parenchyma during the portal phase scanning and more intense during the delayed phase scanning.

OBJECTIVETo investigate the effect of basic fibroblast growth factor (bFGF) and hyaluronic acid (HA) on the proliferation of rabbit chondrocytes in vitro.

METHODSChondrocytes from the knee joints of New Zealand white rabbits were cultured. bFGF or HA or both were added into the culture medium respectively, and the proliferation of the chondrocytes was measured with MTT 3-(4, 5-dimethylthiazol-2-yl) 2, 5-diphenyl-tetra-zolium bromide. (MTT, Sigma, M2128).

RESULTSBasic fibroblast growth factor (10 ng/ml) with low concentration of fetal bovine serum in the culture medium promoted the proliferation of chondrocytes significantly, and this effect reached its maximum when concentration of bFGF reached 50 ng/ml. HA itself had no effect on the proliferation of chondrocytes. However, when bFGF was used in combination with HA, especially when the concentration of bFGF was 50-500 ng/ml and that of HA was 10-50 ng/ml, the effect on the proliferation of chondrocytes was much more than when bFGF or HA was used alone.

CONCLUSIONSbFGF can promote the proliferation of chondrocytes. HA, which has no effect on the proliferation of the cells, can maintain a normal growth of chondrocytes. When bFGF is used in combination with HA, more proliferation is obtained.

Analysis of Variance ; Animals ; Cell Division ; drug effects ; physiology ; Cells, Cultured ; Chondrocytes ; drug effects ; physiology ; Female ; Fibroblast Growth Factor 2 ; pharmacology ; Hyaluronic Acid ; pharmacology ; In Vitro Techniques ; Knee Joint ; cytology ; Male ; Probability ; Rabbits ; Sensitivity and Specificity