BACKGROUND: Islet transplantation is an effective method for the treatment of type 1 diabetes mellitus and parts of type 2 diabetes mellitus. However, its application is hindered by insufficient sources and immunologic rejection. Though transdifferentiation of pancreatic stem cells is at the starting step, it is thought to be the hopeful source for islet cell transplantation.OBJECTIVE: To look for a suitable cells-transplantation source for the treatment of diabetes mellitus. METHODS: The pancreatic ductal epithelial cells were separated from Kunming mice and cultured in DMEM/F12 medium supplemented with keratinocyte growth factor, hepatocyte growth factor and nicotinamide, etc. Samples were taken at different time points for light microscopy and electron microscope. The changes of CK-19 and PDX-1 were detected by immunocytochemistry at 1 and 16 days. The expressions of insulin and glucagon gene were detected by RT-PCR at 1 and 16 days. The physiologic function of these islet-like clusters was determined by dithizone staining and glucose stimulation at 21 days. RESULTS AND CONCLUSION: A large number of epitheliod cells were CK-19 immunoreactive positive and few of them were PDX-1 positive at 1 day after isolation, then CK-19 positive cells proliferated quickly and formed substantial plaques of epithelial cells in cobblestone pattern. At 16 days later, these cells begin to form islet-like clusters gradually, while most of them were PDX-1 immunoreactive positive. The analysis of mRNA by RT-PCR showed very low levels of insulin and glucagon mRNA in the starting materials but increase was found as the process of transdifferentiation. At 21 day differentiated islet-like clusters were stained red by dithizone. In those samples exposed to a stimulatory 15 mmol/L glucose, there was a 1.6-fold increase in insulin compared with to 5.6 mmol/L glucose (P < 0.05). Pancreatic ductal cells of adult Kunming mice could proliferate quickly and have the potency of transdifferentiation into islet-like clusters when cultured in vitro under appropriate conditions.

The process of intervertebral disc degeneration,which could result in intervertebral disc structural and functional change,is a chronic one with multiple factors.The pathophysiologic process is still not completely find out.More and more research reports manifest that certain gene polymorphism also lead to increased risk of intervertebral disc degeneration except environmental factors.Discussions about related genetic factors and their pathophysiological role in the process of degeneration could have a further understanding to disease development.Elucidating genetic components which are associated with degeneration could not only provide insights into the mechanism of the process,but also have clinical significance for early diagnosis and prevention.In order to have a thorough understanding of functional role played by different genes,this paper summarize polymorphism and disease correlation by selecting 15 genes after reviewed the related literature published in recent years.Genetic polymorphisms in 15 genes have been analyzed in association with intervertebral disc degeneration,including aggrecan,collagen Types Ⅰ,Ⅸ and Ⅺ,fibronectin,HAPLN 1,CILP,MMP-1,2 and 3,PARK2,IL-1,6 and VDR.Each genetic polymorphism codes for a protein which has a functional role in the pathogenesis of disease.Among the 15 genes analyzed,polymorphisms in aggrecan,Type Ⅸ collagen,MMP3,IL1,IL6 and VDR show the most promise as functional variants.Genetic studies are necessary for understanding the mechanism of the degeneration.Relevant genetic information could be used as a predictive model for determining individuals' risk for intervertebral disc degeneration eventually.

A double targets of high throughput screening model for xanthine oxidase inhibitors and superoxide anion scavengers was established. In the reaction system of xanthine oxidase, WST-1 works as the probe for the ultra oxygen anion generation, and product uric acid works as xanthine oxidase activity indicator. By using SpectraMax M5 continuous spectrum enzyme sign reflectoscope reflector, the changes of these indicators' concentration were observed and the influence factors of this reaction system to establish the high throughput screening model were studied. And the model is confirmed by positive drugs. In the reaction system, the final volume of reaction system is 50 μL and the concentrations of xanthine oxidase is 4 mU x mL(-1), xanthine 250 μmol x L(-1) and WST-1 100 μmol x L(-1), separately. The Z'-factor of model for xanthine oxidase inhibitors is 0.537 4, S/N is 47.519 9; the Z'-factor of model for superoxide anion scavengers is 0.507 4, S/N is 5.388 9. This model for xanthine oxidase inhibitors and superoxide anion scavengers has more common characteristics of the good stability, the fewer reagent types and quantity, the good repeatability, and so on. And it can be widely applied in high-throughput screening research.

To enhance the quality and efficiency of ozagrel by investigating the differences between the ozagrel polymorphs in bioavailability. Solid ozagrel in different polymorph forms were orally administered to SD rats. An HPLC method was established to determinate plasma level of ozagrel. The bioavailabilities of two polymorph forms were calculated and compared. The pharmacokinetic parameters of ozagrel, were as follows: Cmax was 32.72 ± 17.04 and 34.01 ± 19.13 mg · L(-1), respectively; AUC0-t was 61.14 ± 14.76 and 85.56 ± 18.08 mg · L(-1) · h, respectively; t½ was 1.53 ± 0.51 and 4.73 ± 3.00 h, respectively. There was no significant difference in pharmacokinetic parameters between form I and II polymorphs of ozagrel while the t½ of form II is longer, which indicates that the use of form II polymorph as pharmaceutical product may prolong the effective action time in clinics. This would help the polymorph quality control in drug production.

Objective To evaluate the association between serum 25-hydroxyvitamin D3 [25 (OH) D3],parathyroid hormone,and arterial stiffness in patients with type 2 diabetes.Methods Serum 25 (OH) D3 and parathyroid hormone(PTH) were determined in a cross-sectional sample of 258 patients aged 30 years and over.Arterial stiffness was assessed by pulse wave velocity(PWV) obtained with a VP-1000 pulse wave unit.Fasting plasma HbA1c,lipid profile,calcium,and high sensitive-C reactive protein were determined.Results (1)The prevalence of vitamin D3 deficiency was high(79.84％) in patients with type 2 diabetes.(2) Those with lowered serum vitamin D3 levels had raised PWV [(1610.76 ± 142.70 vs 1527.95 ± 58.02) cm/s,P＜0.05].(3) Multiple stepwise regression analysis showed that 25 (OH) D3 was an impact factor of PWV risk score,which was independent of age,duration of diabetes,and systolic blood pressure(β =-0.256,P＜0.01).(4) Serum PTH was positively correlated with PWV (r =0.210,P ＜ 0.05) and systolic blood pressure (r =0.229,P ＜ 0.05),but negatively correlated with 25 (OH) D3 (r =-0.153,P ＜ 0.05).Conclusions 25 (OH) D3 deficiency is common in patients with type 2diabetes,and a low serum 25 (OH) D3 level is significantly associated with increased arterial stiffness in these patients.The association of serum PTH with arterial stiffness may result via changes in vitamin D and blood pressure.

Objective To evaluate the diagnostic value of triptorelin stimulation test in disorders due to delayed puberty.Methods Triptorelin stimulation test was carried out in 128 teenagers with delayed puberty,due to idiopathic hypogonadotropic hypogonadism(IHH) in 77 cases and constitutional delayed puberty(CDP) in 51.Blood samples were obtained 15 min before and 0,30,60,and 120 min after tripotorelin administration,and the levels of LH and FSH were determined.An extended GnRH stimulation test was carried out in 3 patients with IHH.Results Peak LH,peak FSH,and LH increment were parameters with high diagnostic value.A cut-off point at 8.2 IU/L of peak LH showed a sensitivity of 87％ and a specificity of 80.4％ in the differential diagnosis of IHH and CDP.Conclusion Peak LH cut-off point at 8.2 IU/L of triptorelin stimulation test seems to be sufficient to confirm diagnosis of IHH and CDP.An extended GnRH stimulation test may distinguish hypothalamic from the pituitary hypogonadotropic hypogonadism.

Objective To explore the relationship between Graves disease and hypokalemic periodic paralysis or thymus hyperplasia.Methods Sixty-two patients with Graves disease were enrolled in this study.Thirty-three patients without thymus hyperplasia were selected as group A and other 29 patients were group B.In addition,30 healthy volunteers served as control group.Results There were no significant difference between group A and group B in terms of sex,age,illness course,thyroid enlargement,exophthalmos,periodic paralysis and the levels of free triiodothyronine (FT3),free thyroxin (FT4),thyroid stimulating hormone (TSH) (P ＞ 0.05).The incidence rate of hypokalemic periodic paralysis in male Graves disease with thymus hyperplasia was 62％ (8/13),higher than that in female patients (6％ (1/16),x2 =10.24,P ＜ 0.05).The incidence rate of hypokalemic periodic paralysis in male Graves patients was 62％ (16/26),higher than in female Graves patients (6％ (2/36),x2 =22.96,P ＜ 0.05).There was no significant relationship between Graves disease patients combined with thymus hyperplasia and sex,age,illness course,thyroid enlargement,exophthalmos,periodic paralysis and the levels of FY3,FT4,TSH.Conclusion The incidence rate of hypokalemic periodic paralysis in male Graves disease patients combind with thymus hyperplasia is higher than in female,and therefore it should pay more attention of male Graves patients in case misdiagnosis.

Eleven male patients with pituitary stalk interruption syndrome (PSIS) at our hospital were retrospectively analyzed with respects to clinical features,endocrine status and imaging characteristics.The clinical manifestations included growth retardation and delayed puberty.All of them were deficient in growth hormone and pituitary gonadotropin secretion.Secondary hypothyroidism and hypocortisolism occurred in 8 patients and one had concurrent central diabetes insipidus.MRI with gadolinium contrast revealed no pituitary stalk or slim and anterior pituitary hypoplasia with ectopic posterior pituitary.The clinical characteristics of PSIS are growth retardation and delayed puberty,partial or complete adenohypophyseal dysfunction.The detection of hypothalamic pituitary region by MRI is an important diagnostic tool.

ObjectiveTo analyze and compare the difference and prognosis between vascular embolization and craniotomy occlusion in patients suffering from aneurysmal subarachnoid hemorrhage (aSAH) with Hunt-Hess levelⅢ-Ⅳ, and acute postoperative hydrocephalus.Methods A retrospective study was conducted on 767 patients who had undergone vascular embolization (vascular embolization group,n = 403) or craniotomy occlusion operation (craniotomy occlusion operation group,n = 364), and the patients with postoperative acute hydrocephalus were screened. The clinical data of patients of both groups was analyzed. By judging short-term prognosis in patients with hydrocephalus with Glasgow outcome scale (GOS) score estimated at discharge, the advantages and disadvantages of two surgical procedures were compared.Results The number of cases with postoperative hydrocephalus in vascular embolization group was 56 (13.90%), while that in craniotomy occlusion group was 33 (9.07%). The difference between the two groups of incidence of hydrocephalus was statistically significant (χ2= 4.350,P = 0.037 ). In 767 patients with aSAH, the incidence of hydrocephalus among the patients after the hematoma removal operation was significantly lower than that of patients without hematoma removal [3.07% (11/358) vs. 19.07% (78/409),χ2 = 47.635,P = 0.000]. The incidence of hydrocephalus among the patients after ventricular drainage was significantly lower than that of patients without the drainage [2.77% (19/685) vs. 85.37% (70/82),χ2 = 487.032,P = 0.000]. In 403 cases of vascular embolization group, the incidence of hydrocephalus in the patients after the hematoma removal operation was lower than that of patients without it [8.06% (5/62) vs. 14.96% (51/341),χ2 = 2.082,P = 0.168]. The incidence of hydrocephalus in the patients after the ventricular drainage was lower than that of patients without drainage [2.59% (9/347) vs. 83.93% (47/56),χ2 = 266.599,P = 0.000]. In 364 cases of craniotomy occlusion operation group, the incidence of hydrocephalus in the patients after hematoma removal operation was significantly lower than that of patients did not receive [2.03% (6/296) vs. 39.71% (27/68),χ2 = 95.226,P = 0.000]. The incidence of hydrocephalus among the patients after the ventricular drainage was significantly lower than that of patients without drainage [2.96% (10/338) vs. 88.46% (23/26),χ2 = 203.852,P = 0.000]. The difference in incidence of hydrocephalus between the patients who had hematoma removal surgery between vascular embolization group and craniotomy occlusion operation group was statistically significant [8.06% (5/62) vs. 2.03% (6/296),χ2 = 4.411,P = 0.027], while no statistically difference was present in ventricular drainage patients [2.59% (9/347) vs. 2.96% (10/338),χ2 = 0.085,P = 0.819]. There were 23 patients (41.07%) with good outcome (GOS score 4-5), while 33 (58.93%) with poor outcome (GOS score 1-3) in 56 patients undergone vascular embolization operation. Good result (GOS score 4-5) was shown in 21 (63.64%) and 12 (36.36%) with poor outcome (GOS score 1-3) among 33 patients with hydrocephalus after craniotomy occlusion operation, and the difference was statistically significant (χ2 = 4.230,P = 0.039).Conclusions Hematoma is one of the main factor contributing to the differences in the incidence of postoperative hydrocephalus of Hunt-Hess gradeⅢ-Ⅳ patients either receiving vascular embolization or craniotomy occlusion operation. Lateral ventricle drainage may not be the factor that contributes to the difference in incidence of hydrocephalus formation between the vascular embolization and craniotomy occlusion operation groups in Hunt-Hess levelⅢ-Ⅳ patients. The short term prognosis in the craniotomy occlusion operation group is superior to that of endovascular intervention embolization group.

Objective To explore the relationship between the size and location of the aneurysm after subarachnoid hemorrhage (aSAH) and its clinical classification. Methods A retrospective study was performed in patients with aSAH from January 1, 2008 to December 31, 2014. The relevant clinical data were collected including age, gender, aneurysm size, location, and Hunt-Hess (H-H) classification. The aneurysms were classified by size (A group d<5.00 mm, B group 5.00 mm≤d<10.00 mm, C group d≥10.00 mm), location and H-H classification according to the results of CT, digital subtrac?tion angiography (DSA), and magnetic resonance angiography (MRA). The relationship between size, position of aneurysm and H-H classification was observed and analyzed. Results There were 750 cases included in this study, with average age (56.14 ± 11.88), male 292 and female 458. The total number of aneurysms was 903, and the number of multiple aneurysms was 91 (12.13%). There was one case with multiple aneurysms that can be included in A, B and C groups. There were two cases with multiple aneurysms that can be included in A and B groups, two cases can be included in A and C groups, and three cases can be included in B and C groups. The number of aneurysms and the ratios of groups A, B and C were 20(3.9%), 12 (3.8%), 5 (7.5%), 70 (13.6%), 39 (12.2%), 10(14.9%), 2 (0.4%), 4 (1.3%), 2 (3.0%), 165 (32.0%), 94 (29.4%), 6 (9.0%), 130 (25.2%), 90 (28.1%), 6 (9.0%), 17 (3.3%), 11 (3.4%) and 2 (3.0%) for the location in the anterior cerebral artery, the middle cerebral artery, the posterior cerebral artery, the internal carotid artery, the anterior communicating artery, the posterior communicating artery, and the vertebral basilar artery, respectively. The number of aneurysms and the ratios of H-H classificationⅠ,Ⅱ,Ⅲ,ⅣandⅤin groups A, B and C were 48 (9.3%), 45 (14.1%), 12 (17.9%), 228 (44.2%), 150 (46.9%), 14 (20.9%), 68 (13.2%), 54 (16.9%), 30 (44.8%), 142 (27.5%), 43 (13.4%), 9 (13.4%), 30 (5.8%), 28 (8.8%) and 2 (3.0%). There was a negative correlation between the size of aneurysm and the H-H grade (rs=-0.075, P=0.024). Conclusion The anterior communicating artery and posterior communicating artery are high-risk areas for smaller aneurysms. The internal ca?rotid artery is high-risk areas for larger aneurysms. The size of aneurysm is negatively correlated with H-H classification.