Objective To optimize the radioligand assays for glutamic acid decarboxylase antibody (GAD-Ab), tyrosine phosphatase antibody (IA-2A) and insulin autoantibody (IAA) and to facilitate their application in clinical practice. Methods The radioligand assays established in our laboratory were optimized in several aspects, including the mode and time of antigen-antibody incubation, pH value of TBST buffer, times of washing, and type of centrifuge. The sensitivity and specificity of the optimized assays were calculated by the Diabetes Antibody Standardization Program (DASP, 2003). And the results were compared to those measured with domestic radioimmune assay kits. Results The optimal condition for radiligand assay were as followed: incubating the antigens of GAD-Ab and IA-2A with serum by rotating slowly for 24 hours and IAA for 72 hours respectively; washing the precipitation complex of GAD-Ab and IA-2A with TBST buffer (pH value 7 2～7 4) 4 times and IAA 5 times respectively; and applying the horizontal type of centrifuge. The results from DASP demonstrated that sensitivity and specificity of the optimized assays were 82% and 98% respectively for GAD-Ab, and 64% and 100% for IA-2A. The proportion of consistency between our results for 82 IAA gradient index of optimized assay and those measured with domestic radioimmune kits was 89%. The extensive analysis showed that the consistency reached 100% for those with IAA index less than 0 04 or more than 1 00, and the inconsistency was for the bordline positive ones (index 0 04～1 0). Conclusion The optimized radioligand assays for islet autoantibodies are high sensitivity and specificity, and are applicable in clinical practice.

Unsteadiness and falls in the elderly are a common complaint in primary care and their incidence is expected to increase as the population ages. These patients are physiologically less able to withstand traumatic forces than their younger counterparts and are thus more prone to morbidity and mortality. Aside from intrinsic and extrinsic risk factors, there may be more sinister pathologies underlying these events. We report a case of an elderly gentleman who presented with unsteadiness, clumsiness and recurrent falls. The patient’s son brought to our attention subtle suspicious findings which eventually led to further evaluation. The patient was found to have had bilateral acute on chronic subdural haemorrhages with associated mass effect, underwent emergent decompression surgery and showed marked functional recovery. We review the incidence and outcomes of recurrent falls in the elderly, as well as the incidence, risk factors and varied presentations of subdural haemorrhage in the primary care setting. The case also reminds clinicians of the tendency to overlook and undermanage such patients with seemingly “minor” falls, and the value of family members’ inputs during consultation. It highlights the need for a high index of suspicion and the necessity of lower referral thresholds for further evaluation if indicated.

Electrocardiography ; Humans ; Myocardial Ischemia ; physiopathology

Anemia, Pernicious ; complications ; surgery ; Biopsy ; Endoscopy ; methods ; Humans ; Male ; Middle Aged ; Stomach Neoplasms ; complications ; surgery ; Tomography, X-Ray Computed ; methods ; Treatment Outcome ; Ultrasonography ; methods

Objective To investigate the clinical features of acute mesenteric ischemia (AMI) in the elderly and provide evidence for early prevention,diagnosis and treatment of AMI for elderly patients.Methods A retrospective analysis was performed in 104 patients with AMI in our hospital,who were divided into two groups:the elderly group (aged≥60 years) and the non-elderly group (aged ＜ 60 years).Clinical manifestations,misdiagnosis rate,laboratory data,underlying diseases,treatment and prognosis were compared between the two groups.Results Acute superior mesenteric artery thromboembolus (ASMATE) was the main cause of AMI in the elderly group and acute mesenteric venous thrombosis (ASMVT) was the main cause of AMI in the non-elderly group.Elderly patients were more prone to misdiagnosis as compared with the non-elderly group (P =0.007).Abdominal pain (100.0％) was the most common clinical manifestation of AMI patients,followed by vomiting (58.7％).Nausea and vomiting were strongly associated with AMI in the elderly (P＜0.05).In laboratory data,serum creatinine,lactate dehydrogenase and uric acid were higher and the D-Dimer level was lower in elderly patients than in non-elderly patients (P＜0.05).Hypertension,atrial fibrillation,arteriosclerosis,ischemic heart disease,cerebral infarction and chronic renal insufficiency were more common in the elderly with AMI (all P＜0.05),while history of alcohol intake was higher in the non-elderly group (P=0.042).Hypertension,arteriosclerosis and cerebral infarction were independent relevant factors for elderly-onset AMI (P =0.000,OR =4.057;P =0.001,OR=4.585;P=0.007,OR=4.269).The number of patients with colon necrosis was higher in the elderly group than in the non-elderly group (P=0.038),and the prognosis was worse in the elderly group than in the non-elderly group (P=0.001).Conclusions ASMATE is the main cause of AMI in elderly patients.Nausea and vomiting are the main clinical manifestations,together with impaired renal function.Hypertension,arteriosclerosis and cerebral infarction are the independent relevant factors for elderly-onset AMI.Elderly patients are more prone to misdiagnosis,show more severity and have worse prognosis.

To construct a cDNA subtractive library of genes transactivated by hepatitis C virus core protein with suppression subtractive hybridization technique. mRNA was isolated from HepG2 cells transfected with pcDNA3 1(-)-core and pcDNA3 1(-) empty vector,respectively, then cDNA was synthesized. After restriction enzyme RsaI digestion, small sized cDNA were obtained. Then tester cDNA was divided into two groups and ligated to the specific adaptor 1 and adaptor 2, respectively. After tester cDNA was hybridized with driver cDNA twice and underwent two times of nested PCR, and then it was subcloned into T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E.coli strain JM109. The cDNA were sequenced and analyzed in GenBank with Blast search after PCR. The subtractive library of genes transactivated by HCV core protein was constructed successfully. The amplified library contained 233 positive clones. Colony PCR product showed that 213 clones contained 100~ 1 000 bp inserts. Sequence analysis was performed in 63 clones. Six of the sequences were unknown genes before. The full length sequences were obtained with bioinformatics method,which had been accepted by GenBank. It suggested that six novel cDNA sequences might be target genes transactivated by HCV core protein.

Objectives To explore the effect and advantages of seperatable nylon ring ligation under endoscope for esophageal varices hemorrhage.Methods 24 cases of esophageal varices hemorrhage accepted ligation treatment under endoscope guide by using Olympus separatable nylon ring ligation apparatus. The effect was recorded. Results The average ligation points on one case were 10 2. The emergency hemostatic rate hemorrhage from esophageal varices repture was 100%. The disappearance rate of esophageal varices was 73 7%. No severe complication was occurred during operation.Conclusions Seperatable nylon ring ligation treatment under endoscope is a new safe and effective method for esophageal varices hemorrhage. This method is simple and cheap.so it has valuable for spreading use in clinic.

AIMTo develop a rat model of insulin resistanc e,and to study the effect of puerarin injection on expression of protein kinase B in insulin resistanc e rats. METHODS30 SD rats were randomly divded into two groups. The model group were given the high fat diets(0 30 pig fat, 0 10 sucrose, 0 0 3 cholesterol) for 8 weeks. Then 9 rats of the group had been choosen as the pu erarin treatment group and were given abdominal injection (100 mg?kg -1 ?d -1 ) for 4 weeks. By the end of the experiment, Western blot and computer i mage pattern analyze system were used to analysis expression of PKB in skeletal muscles. RESULTS(1) The model group showed a hyperglycemia, hype rinsulinism and obviously visceral obesity by high fat-feeding, the insulin res istance index (ISI) decreased while the HOMA insulin resistance index (HOMA-IR) increased compared to the normal group, insulin resistance was induced in this way; (2) Compared with the pathology control group:fasting blood glucose (FBG), fasting insulin (FINS), visceral fat mass and the HOMA insulin resistance index (HOMA-IR) of the treating group were significantly decreased whereas the insuli n index (ISI) enhanced. Expression of PKB was markedly increased by puerarin tre atment. CONCLUSIONPuerarin injection can significantly elevate e xepression of PKB and ameliorate the state of insulin resistance. Improved biolo gic effect of insulin and prevention of the deleterious effect of fat may be in volved in the mechanism concerned.

Wnt/β-catenin signaling pathway plays an important role in the proliferation, growth, invasion, and metastasis of human cancers. Moreover, β-catenin/T-cell factor 4 (TCF4) interaction regulates the transcription of the key oncogenes in Wnt/β-catenin signaling pathway. Therefore, β-catenin/TCF4 interaction would be a promising therapeutic target for the development of highly selective anticancer agents. At present, most ongoing small-molecule inhibitors targeting β-catenin/TCF4 interaction, including PKF222-815, iCRT3/5/14, LF3, and sanguinarine, have been developed in preclinical studies for human cancer therapeutics. In this review, we summarized the research advances of up-to date inhibitors targeting β-catenin/TCF4 interaction, including the molecular structure and cellular functions of β-catenin in canonical Wnt signaling pathway. This review holds a hopeful avenue for the development of novel and highly selective Wnt inhibitors targeting β-catenin/TCF4 interaction for future anticancer strategy.

ObjectiveTo investigate serum lipopolysaccharide (LPS) level in people with different glucose tolerances and to explore the relationship between LPS and insulin resistance/β-cell secretory function.Methods Sixty-seven subjects were recruited,including 23 with newly diagnosed type 2 diabetes ( T2DM),21 impaired glucose tolerance ( IGT),and 23 normal glucose tolerance (NGT).Serum LPS was assayed by limulus amebocyte lysate test ;expression of LPS toll-like receptor 4 (TLR4) on surface of plasma monocytes was measured by flow cytometric assays,and the changes of LPS levels by 0.5 hours and 2 hours after a high-fat diet were detected.Insulin resistance was evaluated by homeostasis model assessment for insulin resistance (HOMA-IR); β-cell secretory function was evaluated by homeostasis model assessment for β3 cell function ( HOMA-β )/HOMA-IR,increment in insulin in the first 30 minutes/increment in glucose in the first 30 minutes ( AIns30/ΔG30)/HOMA-IR,AUCIns120min/HOMA-IR.Results2 h LPS levels after a high-fat diet were significantly higher than fasting LPS levels [ NGT:0.96(0.33,0.99)vs 0.62 (0.22,0.64),IGT:1.08(0.53,1.22)vs 0.71 (0.39,0.82),T2DM:1.23 (0.62,1.43)vs 0.86( 0.45,0.94 ),EU/ml,all P＜0.01 ].Fasting,0.5 h,and 2 h LPS levels and fasting TLR4 levels of T2 DM group and IGT group were respectively higher than those of NGT group [ fasting LPS:0.86( 0.45,0.94 ),0.71 ( 0.39,0.82 ) vs 0.62(0.22,0.64),EU/ml;0.5 h LPS:1.10(0.55,1.18),0.84(0.50,1.07) vs 0.73(0.31,0.76),EU/ml;2 h LPS:1.23(0.62,1.43),1.08(0.53,1.22)vs 0.96(0.33,0.99),EU/ml; fasting TLR4:36.96( 17.22,55.19),30.34 ( 15.00,45.18 )vs 15.66 (6.09,9.76),MIF/105 cells,all P＜0.01 ].Fasting LPS,AUCLPS 120 min,and fasting TLR4 were positively correlated with insulin resistance index and negatively correlated with β-cell secretory function index ( P＜0.05 ).Multiple linear regression analysis showed that fasting LPS was an independent correlative factor of HOMA-IR and 0.5 h LPS was an independent correlative factor of (AIns30/AG30)/HOMA-IR and AUCIns Ins120min/HOMA-IR.ConclusionPeople with different glucose tolerances show differed LPS levels and its receptor TLR4 levcls,both of which are correlated with insulin resistance and β-cell secretory function,suggesting that LPS is associated with the pathogenesis of abnormal glucose regulation.