Objective To investigate the effect of alpha 1-antitrypsin (A1AT) concerning in reducing the injury of transplanted islets by pancreas exocrine cells and promoting proliferation of the pancreas B cells.Method The pancreases of mice were digested with collagenase,islets were isolated artificially,and pancreatic exocrine cells were collected.In purified islet group (n =6),100 islets were seeded into a 6 well culture plate.In experimental group(n =6),100 islets were co-cultured with equal volume of pancreas exocrine cells,and 0.5 mg/mL A1AT was added into a 6-well culture plate.In control group(n =6),100 islets were co-cultured with equal volume of pancreas exocrine cells.After 48 h,insulin content of islets in each well and trypsin concentration in the supernatant of each well were measured.The islets were cultured in low sugar and high sugar 1640 medium,then glucose stimulated insulin secretion (GSIS) test was carried out.In vivo,8-9-week old male BALB/C mice were induced with STZ (190 mg/kg body weight,i.p) to establish the diabetic model and randomly divided into two groups.In experimental(n =10) and control(n =10) groups,250 islets and the equal volume of pancreatic exocrine cells were transplanted into different regions of left kidney subcapsule,resepctively.The experimental group was injected with A1AT (83 mg/kg,qd,i.p) for 28 days after operation,and the control group was injected with the same amount of normal saline (qd,i.p) for 28 days.Both two groups were given EDU (5 μg/g,qd,i.p) for 28 days.The blood glucose level was monitored continually.Nephrectomies were performed after 28 days.The expression of anti-amylase antibodies in the renal subcapsule was detected by immunohistochemical staining,and the proliferation of islet beta cells was examined using immunofluorescence staining.Result Insulin levels and insulin stimulation index in the control group were decreased as compared with those in the purified islet group; those in the experimental group were higher than in the control group,but lower than in the purified islet group.Trypsin concentration in the control group was increased as compared with the purified islet group,that in the experimental group was lower than the control group,but higher than in the purified islet group (all P＜0.01).After islets transplantation,the blood glucose levels in control and experimental groups were normal,but those in the control group recovered later than in the experimental group (P＜0.01).At 3rd day after nephrectomy,the blood glucose levels were ＞21 mmol/L in both two groups.A large number of anti-amylase antibody-positive cells were found in the renal subcapsule in the control group while little seen in the experimental group after 28 days.The immunofluorescence showed that the insulin +/EDU + B cells in the experimental group were more than those in the control group.Conclusion Conclusion Co-culture of islets and pancreatic exocrine cells with A1AT can prevent islet cells from damage caused by trypsin.A1AT could inhibit the secretion of pancreatic amylase from pancreatic acinar cells and promote proliferation of islet beta cells.

OBJECTIVE: To evaluate the value of T-SPOT.TB in the diagnosis of tuberculous peritonitis (TBP). METHODS: A total of 50 patients with clinically suspected TBP admitted from August 2011 to July 2012 from the Third Xiangya Hospital were enrolled in this prospective cohort study. Peripheral blood T-SPOT.TB, purified protein derivatives of tuberculin (PPD) skin test, serum tuberculosis antibody (TB-Ab) and ascitic adenosine deaminase (ADA) were measured in the 50 patients and we compared the diagnostic value of the 4 methods. RESULTS: According to the standard of diagnosis of TBP and grouping, 24 patients were diagnosed with TBP, 17 non-TBP and 9 undiagnosed in the end. The sensitivity of T-SPOT.TB for the diagnosis of TBP was 91.7% (22/24), with statistical significance when compared with PPD skin test 37.5% (9/24), serum TB-Ab 16.7% (4/24), and ascitic ADA 36.4% (8/22) (P<0.01). The specificity of T-SPOT.TB for the identification of non-TBP was 76.5% (13/17), without statistical significance when compared with PPD skin test 70.0% (7/10), serum TB-Ab 78.6% (11/14), and ascitic ADA 100% (14/14) (P>0.05). The positive prediction value of T-SPOT.TB for the diagnosis of TBP was 84.6% (22/26), without statistical significance when compared with PPD skin test 75.0% (9/12), serum TB-Ab 57.1% (4/7), and ascitic ADA 100%(8/8) (P>0.05). The negative prediction value of T-SPOT.TB for the identification of non-TBP was 86.7% (13/15), with statistical significance when compared with PPD skin test 31.8% (7/22), serum TB-Ab 35.5% (11/31), and ascitic ADA 50.0% (14/28) (P<0.05). CONCLUSION Peripheral blood T-SPOT.TB for the diagnosis of TBP is highly sensitive, which is better than PPD skin test, serum TB-Ab, and ascitic ADA. T-SPOT.TB has an important reference for diagnosing suspected TBP quickly and accurately.
Adolescent ; Adult ; Aged ; Child ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Humans ; Interferon-gamma Release Tests ; methods ; Leukocytes, Mononuclear ; immunology ; Male ; Middle Aged ; Peritonitis, Tuberculous ; diagnosis ; Sensitivity and Specificity ; T-Lymphocytes ; immunology ; Young Adult