Objective To investigate the interrelation between microtubule associated protein 5 and lysosome in cultured spinal neurons. Methods Neuron culture, immunofluorescence cytochemistry, laser confocal scanning microscope were used. Results In cultured spinal neurons, both of MAP5 and calthepsin D were distributed in cytoplasm and processes. In merged pictures, the distribution of MAP5 and calthepsin D was almost similar. When nocodazole was employed, the distribution of MAP5 and calthepsin D changed simultaneously. Both fluorescence distributed in cytoplasm was uneven and the process was shorter. When PMA was employed, the distribution of red and green fluorescence became reticulum and spread toward membrane. The soma enlarged and the process displayed certain direction accompanying adjacent processes interlinked.Conclusion In cultured spinal neurons, the distributions of MAP5 and lysosomes were almost similar. Both were in cytoplasm and process. Nocodazole and PMA caused reverse change for the distribution of MAP5 and lysosomes. The distribution of MAP5 and calthepsin D treatel with Nocodazole was unregulated, while PMA was regulated and loose. These results suggested that MAP5 and lysosomes were interrelated between structure and function. Both of them were depended on the integrity of microtubule.

ObjectiveTo observe the changes of phosphorylated extracellular signal-regulated kinase 1/2 (pERK1/2) and c-fos in the medial prefrontal cortex (mPFC) of post-traumatic stress disorder (PTSD) rats. Methods Male Wistar rats were randomly divided into control group and PTSD model group. The model group rats were exposed to the single-prolonged stress (SPS) to set up the rat PTSD model. The expression of pERK1/2 was detected using immunohistochemistry and Western blotting, and the expression of c-fos mRNA was detected using reverse transcription-polymerase chain reaction (RT-PCR). Results The result of immunohistochemistry analysis showed that the number of pERK1/2-positive cells of control group and model group were 10.4±2.07 and 48.8±10.08 respectively (P<0.01), and integral optical density were 24.955±3.691 and 110.810±10.643 respectively (P<0.01). And Western blotting showed that relative expression quantities of pERK/β-actin were 0.510±0.052 and 1.109±0.106 respectively (P<0.01). The quantities of c-fos mRNA relative expression of control group and model group were 0.267±0.067 and 1.049±0.131 (P<0.01). Conlusion The levels of pERK1/2 and c-fos increase significantly in mPFC of PTSD model rats. The ERK signal transduction pathway in mPFC might play an important role in the pathogenesis of PTSD.

Objective To evaluate the effect of the needle free injection system (INJEX30) and insulin pen on insulin absorption and glycemic control in diabetic patients.Methods A total of 30 diabetic patients on insulin therapy without obvious complications were enrolled in the study with average BMI of 25.24 kg/m2.A comparison study was carried out in those subjects with the INJEX30 and insulin pen at 1 st day and 5th day.After an overnight fasting of 8-10 h,a standard mixed meal(50 g bread,50 g egg and 250 ml milk) was given to each patient.Blood samples at 0,20,40,60 min of the standard mixed meal were collected to test plasma glucose,serum insulin and C peptide.Results No difference was shown in fasting plasma glucose,serum insulin and C peptide between the patients with the two injection methods.The area under the curve (AUC) of plasma glucose and serum C peptide was significantly lower after the INJEX30 injection than that after insulin pen injection [plasma glucose AUC (542 ± 172) min · mmol · L-1 vs (601 ±199) min· mmol · L-1,P ＜0.01; C peptide AUC (70 ±53) min · μg · L-1 vs (80 ±58) min · μg · L-1,P ＜0.01].The AUC of serum insulin was significantly higher after the INJEX30 injection than that after insulin pen injection [serum insulin AUC(5621 ± 3790) min · mIU · L-1 vs(4285 ± 3376) min · mIU · L-1,P ＜0.01].No difference was found in the AUC of serum insulin between the two injection methods in the patients with BMI below 25.24 kg/m2,while the AUC of serum insulin was significantly higher after the INJEX30 injection than the insulin pen injection in the patients with BMI above 25.24 kg/m2 [serum insulin AUC(6453 ± 4099) min · mIU · L-1 vs (4879 ± 3701) min · mIU · L-1,P ＜0.01].Conclusion The INJEX30 improves the serum insulin level which may lead to a beneficial effect on the glycemic control.Such effect is more obvious in the overweight patients.

Objective: To research effect of different doses of Changle on the structure and function of rat liver.Methods: 180 healthy Wistar rats were divided into 5 experimental groups according following doses: 0.1,0.2,2.0,10.0 and 20.0 mg*kg-1,respectively, and the control group given physiological saline for six months. The changes of liver structure were examined by means of normal histological chemistry and transmission electron microscope(TEM). Results: The body weight of animal was linearly increased with the decrease of administered doses, gradual reduction of glycogen in hepatocytes and infiltration of inflammatory cells in the portal area were found in the group of 20.0 mg*kg-1. Changes of ultrastructure showed there were dense bodies and lysosomes containing dense granules in Kupffer cell and hepatocyte,and they were increased along with doses adding. Nuclei deformed, ALP and GPT in serum were rose in the group of 20.0 mg*kg-1. Different doses of Changle could lead to distinct biological effects. Conclusion: Long-termed administration of 20.0 mg*kg-1 Changle can lead to damage of structure and function of rat liver.

Objective The aim of this study is to identify association between increased serum activity of alanine aminotransferase (ALT) and metabolic parameters in non-alcoholic population of north China. Methods A total of 5351 subjects who came to visit Yuquan Hospital of Tsinghua University for health examinations during May to December 2007 were divided into two groups based on their serum ALT activities, one with equal to or more than 40 U/L and the other with less than 40. Anthropometric data, blood pressure, serum ALT, serum levels of triglyceride (TG), total cholesterol (TC), high-dense lipoprotein-cholesterol (HDL-C), low-dense lipoprotein-cbolesterol (LDL-C) and fasting blood glucose (FBG) were measured for all of them. Results Body mass index (BMI), systolic blood pressure (SBP), diastolic blood pressure (DBP), TG, TC, HDL-C, LDL-C and FBG were significantly different between the two groups. BMI, TG, TC, HDL-C, LDL-C and FBG all associated with increased ALT both in males and females. DBP associated with increased ALT in males and SBP with it in females, logistic regression analysis showed that male, younger age, raised BMI, TG, TC, FBG and reduced HDL-C were strongly associated with increased serum ALT. Conclusions Results showed that metabolic disturbance such as obesity, hypertension, dyalipidemia, hyperglycemia, and so on associated with increased ALT in non-alcoholic population of north China.

Objective To detect protein PTEN in live and muscle tissue of KKAy diabetic mice and to investigate whether PTEN is associated with the insulin resistance in diabetic KKAy mice.Methods Animals were divided into normal diet C57BL group(n=7,sixteen-week-old C57BL mice),high fat diet C57BL group(n=7),diabetic KKAy group(n=7),the latter two groups were fed with normal diet until week 12,followed by high fat diet for 4 weeks.Blood glucose was measured every week,it would be diagnosed of diabetes if blood glucose more than 300 mg/dl(16.7 mmol/L) in two consecutive weeks.Dispatched the mice,took quadriceps of femoris muscle and liver tissue,added tissue lytic solution,measured the protein concentration by Bradford method and detect the PTEN protein expression in muscle and liver tissue by Western bolts methods.Results The PTEN protein level was significantly increased in quadriceps muscle and live tissue in KKAy mice as compared to the both age-matched C57BL control groups(P

Objective To construct and to identify eukaryotic expression vector expressing Islet-brain 1(IB1) gene.Methods Total RNA was extracted from human insulinoma.IB1 gene was amplified by PCR from human IB1cDNA library.The eukaryotic expression vector encoding IB1 was constructed by inserting the IB1 cDNA into EcoR I/Kpn I sites of the pEGFP-N1 vector with the green fluorescent.The construct was transfected into RINm5F cell line,screened by G418.The phase contrast fluorescence microscope,flow cytometer,and Western blot were used to identify the recombinant plasmid and transfeced cell line.Results The RT-PCR products for IB1(AA1-280)generated from human insulinoma was 840 bp.Sequence analysis proved the same sequence as published in Gen-Bank.Two bands showed that pEGFP-N1 vector encoding IB1 digested by EcoR I or Kpn I.Western blot showed IB1 gene was expressed in RINm5F cells.Conclusion The recombinant prokaryotic expression plasmid pEGFP-N1-IB1 has been successfully constructed.

Objective To detect the expression of FoxO3a in adipose tissue from KKay diabetic mice and the effects of treatment with rosiglitazone and metformin on FoxO3a expression in adipose tissue in order to understand the mechanism of insulin resistance.Methods Mice were randomly divided into 3 groups: the group without any treatment,group with rosiglitazone and group treated with metformin 3 g/kg/day.Control group consists of 7 C57BL mice 16.Dispatched the mice and sampled adipose tissue to measure the protein concentration by Bradford method and to detect Foxo3a protein expression on adipose tissue by Western Blot with multiple colony antibody 1∶1250.Results Foxo3a was highly expressed in adipose from KKAy diabetic mice as compared with that in control group(FoxO3a/?-actin ratio 1.76?0.19 vs 1.15?0.10,P

Objective: Endoscopic measurement mainly depends on visual estimation or open biopsy forceps,which have been proved lacking in accuracy due to the image barrel distortion caused by wide-angle lens in endoscope.This study aimed to validate a new concept of "virtual internal standard" proposed by ourselves and develop a new method for endoscopic measurement based on the concept.Methods: We photographed planar grid graphs printed with standard calibrations with an electronic endoscope at different object distances,obtained the information on the virtual internal standard from each photo at the corresponding object distance,and analyzed the digital characteristics of the barrel distortion photos,including the size of the visual field,the image compression ratio,and the credible area for measurement with the virtual internal standard.Based on the analyses,the method of endoscopic measurement by the virtual internal standard was established and experiments both in vivo and in vitro were conducted.Results: The ratio of the object distance to the visual field was 1 ∶ 2 when the former was between 10 mm and 40 mm.Calculation of the compression ratio showed that the credible area of measurement in the central area was 10 mm at the object distance of 10-15 mm,20 mm at 20-30 mm,and 30 mm at 30-50 mm.The credible area of measurement on various planes accounted for 2/5 to 3/5 of the total image width.There was a tendency of double-curve change in the arrangement of pixel values of the central grids on different planes.The differences between the pixel values per centimeter became slight at the object distance of 35-50 mm.Conclusion: Endoscopic measurement by the virtual internal standard is a objective,accurate,simple and practical method.

Objective:To analyze the delays in publication of articles (DPA) in the Journal of Medical Postgraduates(JMP).Methods:According to the corresponding rules for monthly academic journals(national:DPA