Objective To explore the inhibition mechanism and safety of pyrazolo[1,5-a]pyrazin-4(5H)-one derivatives against proliferation of human lung cancer A549 cells, H322 cells, and human umbilical vein endothelial cell (HUVEC). Methods Cells were treated with 40μmol/L of the ppo3a, ppo3b, ppo3i, and 0.1% DMSO (control) for 48 hours, respectively. Apoptosis was determined by Hoechst 33258 staining assay in H322 and A549 cells. Cell cycle distribution was determined by flow cytometry analysis in A549 cell. LC3-II, p53, and heat shock protein (HSP) 70 protein levels were detected by Western blotting in A549 cells treated with ppo3b for 48 hours. The morphology and viability of HUVEC were observed by inverted microscope and sulforhodamine B (SRB) assay. Results Ppo3a, ppo3b, and ppo3i significantly induced apoptosis in H322 and A549 cells. A strong G1-phase arrest was concomitant with the growth inhibitory effect on A549 cells. Ppo3b effectively elevated the p53 protein level, but significantly reduced the HSP70 protein level. There were no significantly inhibitory effect on the morphology and viability of HUVEC when treated with ppo3a, ppo3b, and ppo3i. Conclusions ppo3a, ppo3b, and ppo3i could inhibit H322 proliferation through apoptosis and inhibit A549 through apoptosis and G1-phase arrest. The protein p53 and HSP70 might involve in the inhibition effects. These derivatives might be a clue to find effective and safe drug for lung cancers.

Objective:To search for a method for radical resection of pancreatic and duodenal malignancy involving the mesentery root and for the long post-operation survival of patients.Methods:From Jan.2004 to Aug.2006,a total of 26(16 male and 10 female. aged 27-70)patients with pancreatic and duodenal malignancy involving the mesentery root were treated in our department.The patients included 3 with duodenal malignancy and 23 with pancreatic malignancy.Curative resection was performed by the extended pancreaticoduodenetomy(Whipple procedure)combined with mesentery root resection(MRR)for all patients.The outcomes,safety and the post-operation survival rate were analyzed retrospectively.Results:Thirteen patients were treated with Whipple procedures combined with MRR,9 were treated with partial portal vein/superior mesenteric vein(PV/SMV)and reconstruction of the vessel,and 4 patients received pre-shunt between PV and SMV with artificial vessel graft before the extended Whipple and MRR procedures.The operation time was 2.5 to 7(4.4?1.1)hour,and blood loss was 300 to 5 000(1892?1414)ml with the blood transfusion of 0 to 5 600(2 100?1 586)ml.There was no death in our group and 7(27%)had post-operation complication.The post-operation hospital stay was 10 to 30 days.The pathologic examination showed negative surgical margins for all specimens.The tumor size was 4 to 10 (6.17?2.03)cm.After a follow-up of 9 to 38 months,the pain was relieved in all patients.One of the 3 patients with duodenal adenocarcinoma had liver metastasis at 10 months after operation,and the other 2 survived 10 months and 27 months without evidence of tumor reccurence.The patient with pancreatic micro-adenocarcinoma died of local reccurence 9 months after operation.The patient with neuroendocrine carcinoma died of organ failure 24 months after operation.The patient with lymphoma have survived for 24 months after operation.The 1-year and 2-year accumulated survival rates in the 20 cases with pancreatic ductal cancer were 86.6% and 45.6%. respectively.Conclusion:The extended Whipple procedure with MRR is safe and effective.It can obtain R0 resection in patients with malignant tumors(over 5 cm in diameter)in the head,neck and uncinate process of the pancreas and duodenal.

OBJECTIVETo investigate the growth and development of nerve growth factor (NGF)-positive neurons in the cerebellum of midanaphase human fetus.

METHODSThe expression of the NGF-positive neurons in the cerebrum of human fetus was observed by immunohistochemical methods, and the integral absorbance (IA) was detected.

RESULTSBy the 3rd to 4th month of gestation, neurons was seen in the ependymal, central, and marginal plate of cerebellum; the nucleus was oval and the neurons had short and small processes. By the 5th to 7th month of gestation, the number of NGF-positive neurons increased, the expressions enhanced, the nucleus was round-, oval-, or fusiform-shaped, the neurons grew larger in size, and the Purkinje cells showed NGF-positive expression. By the 8th to 10th month of gestation, the NGF-positive expression was enhanced with deeper dying, the body of Purkinje cells grew larger gradually, and the number of NGF-positive neurons in the granular cell layer and molecular layer increased. IA of the cerebellar cortical neurons of the 3rd, 4th, 5th, 6th, 7th, and 8th month of gestation showed an increasing trend, and significant difference was observed (P < 0. 05).

CONCLUSIONNGF-positive neurons in the cerebellum play an important role for differentiation, proliferation, migration, and growth of neurons in the cerebellum.

Cerebellum ; cytology ; metabolism ; Fetus ; cytology ; metabolism ; Humans ; Nerve Growth Factor ; metabolism ; Neurons ; cytology ; metabolism ; Purkinje Cells ; metabolism

OBJECTIVETo study the value of MRI in the diagnosis of cerebral sparganosis in children.

METHODSThe MRI appearances of 17 children with cerebral sparganosis proven by pathology or serological test were retrospectively studied. The diagnostic accuracy rate of cerebral sparganosis was compared before and after knowing the imaging features of this disease.

RESULTSEnhanced MRI was performed in 12 cases, and all of them demonstrated abnormal enhancement. Peripheral ring-type, tortuous beaded shape or serpiginous tubular shape enhancement were found in 8 cases. After analysis of follow-up MRI for 7 cases, a change in location and shape of lesions was found in 2 cases. Typical MRI patterns of cerebral sparganosis included abnormal enhancement such as peripheral ring-type, tortuous beaded shape or serpiginous tubular shape enhancement, and changes in location and shape of lesions in the follow-up MRI. The diagnostic accuracy rate for the 17 cases of cerebral sparganosis at the first visit, after first imaging examination and after several follow-up MR examinations were 0%, 11.8% and 28.6%, respectively. The diagnostic accuracy rate of this disease increased to 64.7%, 70.6% and 85.7% respectively after the radiologists were trained in the imaging characteristics of the disease.

CONCLUSIONSMR findings in children with cerebral sparganosis are specific. Mastering correctly the typical imaging features of the disease can be very helpful in reducing the rate of clinical misdiagnosis.

Adolescent ; Central Nervous System Helminthiasis ; diagnosis ; pathology ; Child ; Child, Preschool ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Retrospective Studies ; Sparganosis ; diagnosis ; pathology

OBJECTIVETo compare profiles and related factors of irregular bleeding induced by different types of low-dose hormone therapy (HT) and a Chinese formulated herbs products.

METHODSApplied with open-labeled, randomized, and clinical trial design, 136 postmenopausal women were assigned into four groups: group A: estradiol valerate (E2 V) 1 mg/d + medroxyprogesterone acetate (MPA) 2 mg/d; group B: conjugated equine estrogen 0.45 mg/d + MPA 2 mg/d; group C: tibolone 1.25 mg/d; group D: a Chinese formulated herbs product (Kuntai) 4# tid. Each subject took element calcium 400 mg/d and vitamin D 200 IU/d concomitantly. Modified Kupperman scores were assessed on baseline and every 3 months thereafter and irregular bleeding was recorded on menopausal diary every day. The duration of this study was 1 year. Results The efficacies were similar in three HT-managed groups, but was better than in group D, although the latter was also effective in alleviating menopausal symptoms. Hazard ratio (HR) of irregular bleeding was 1.00 in group C, 2.43 in group A (95% CI: 1.08-5.46), 3.12 in group B (95% CI: 1.42-6.88), and 0.73 in group D (95% CI: 0.26-2.04). Most cases initially experienced bleeding in the first 3 months but such initiation was a bit later in group C. Endometrium, as detected by B-mode ultrasound, increased approximately 1 mm in HT groups, while it was a bit thicker in group C. Long periods in reproductive age and short time since menopause were high risk factors for irregular bleeding.

CONCLUSIONProfiles of irregular bleeding in 3 commonly used types of low-dose HT are different and some factors such as long period in reproductive age and short time since menopause may contribute to bleeding initiation.

Adult ; Aged ; Double-Blind Method ; Estradiol ; administration & dosage ; analogs & derivatives ; Estrogen Replacement Therapy ; adverse effects ; Estrogens, Conjugated (USP) ; administration & dosage ; Female ; Humans ; Medroxyprogesterone Acetate ; administration & dosage ; Metrorrhagia ; etiology ; Middle Aged ; Norpregnenes ; administration & dosage ; Phytotherapy ; adverse effects ; Postmenopause ; Risk Assessment

OBJECTIVETo investigate the development of nitric oxide synthase (NOS)-positive neurons in the frontal lobe of the cerebrum of human fetus in midanaphase.

METHODSThe positive expression of the NOS-positive neurons in the frontal lobe of cerebrum of human fetus was observed by immunohistochemistry.

RESULTSBy the 7th to 8th month of gestation, NOS-positive neurons in the cortical plate of frontal lobe demonstrated themselves inequality of sizes and morphological difference in the deeper layers with interspersed distribution and increased NOS response, and the distribution of beaded nerve fiber was observed between neurons of cerebral tissues. By the 9th to 10th month of gestation, NOS-positive neurons in the deeper layers of cortical plate of frontal lobe developed slightly in size of the cell body with richer cytoplast, full shape and deeper dyeing and extrusive beaded nerve fibers, and the NOS-positive neurons scattered in the shallow layer of cortical plate presented with round or oval shape. The nucleus developed bigger but with sparse cytoplasm and clear nerve process.

CONCLUSIONNOS-positive neurons in the deeper layer of cortical plate of lobus frontal consist of largely network of neural system and produce micro-environment with higher concentration of NO, which favors the differentiation, proliferation, migration, and development of various neurons.

Cerebrum ; cytology ; embryology ; Female ; Fetus ; cytology ; enzymology ; Frontal Lobe ; cytology ; embryology ; Humans ; Immunohistochemistry ; Neurons ; cytology ; enzymology ; Nitric Oxide Synthase Type I ; metabolism ; Pregnancy

Objective: To investigate whether insulin stimulates the translocation of glucose transporter-4 (GLUT4) and glucose uptak e in ischemic myocardium. Methods: Plasma concentration of gluc ose, lactate, free fatty acid and insulin were determined by autoanalyser, and G LUT4 was studied by Western blotting analysis. Results: Insulin increased GLUT4 significantly in sarcolemma of ischemic myocardium ［(25±4)% vs (40±6)%］, and GLUT4 content in intracellular membrane decreased proporti onally. The glucose uptake increased significantly in insulin-ischemic myocardi um. The uptake of insulin-ischemic myocardium was almost 2 times that of ischem ic myocardium. Conclusion: Insulin stimulation results in GLUT4 translocation and increases glucose uptake in ischemic myocardium. When myocardi al ischemia occurs, insulin is helpful in increasing myocardial glucose uptake a nd utilization.

Objective: To investigate whether there is additi ve effects of hyperinsulinemia and ischemia on expression of canine myocardial G LUT4 gene in vivo. Methods: The expression of myocardial GLU T4 was determined by semiquantitative immunoblotting.The expression of GLUT4 mRN A was determined by semiquantitative Northern blotting. Results: Dramatic changes were seen in GLUT4 mRNA and GLUT4 expression in the ischemic hearts.After infusing insulin for 8 h,regional GLUT4 mRNA and GLUT4 levels in is chemic hearts were 2.5, 2.3-fold that of expression in normal hearts(P<0.01 ). Myocardial glucose uptake in ischemic hearts was increased by 4-fold when co mpared with normal hearts(P<0.01). Conclusion: There are not only additive effects of hyperinsulinemia and low-flow ischemia on canine myoc ardial GLUT4 mRNA and GLUT4 expression in vivo, but also increase of myocar dial glucose uptake. Enhanced GLUT4 expression may be an important protective m echanism by which myocardial cells enhance glucose uptake and metabolism during low-flow ischemia.

The complete nucleotide sequence of a cDNA clone encoding weever ribosomal protein L8, WeevL8, is described here. The WeevL8 cDNA has 848 nucleotides in full length and encodes a 257 amino-acid protein with a calculated molecular mass of 28.02 kDa. Two conserved domains have been identified in WEEVL8. It was concluded fromsequence alignment that WeevL8 gene was quite conservative, consistent with its role as a house-keeping gene. A phylogenetic analysis made by L8 protein showed the similar phylogenetic relationship to that with 18s rDNA. The high similarity supports the notion that ribosomal protein L8 can also be used as phylogenetic criterion.

OBJECTIVETo investigate the growth and development of nitric oxide synthase (NOS)-positive neurons in the cerebellum of human fetus in the midanaphase.

METHODThe positive expression of the NOS-positive neurons in the cerebellum of midanaphase human fetus was observed by immunohistochemistry.

RESULTSBy the sixth to seventh month of gestation, NOS-positive neurons were seen in the ependymal layer of the cerebellum. The nucleus was oval-shaped and the neurons had short and small processes. By the eighth to ninth month, NOS-positive neurons were found in the central layer of the cerebellum and the nucleus was round-, oval-, or fusiform-shaped; meanwhile, the neurons grew larger in size with richer cytoplast and heavier staining. The beaded nerve fibers reached the marginal layer and the layer became thickened on the tenth month, which generally was composed of 5 to 6 layers of NOS-positive neurons that were tightly aligned. Some NOS-positive neurons were in smaller size with the cell body and the nerve fibers grew well.

CONCLUSIONNitric oxide generated by NOS of the NOS-positive neurons in the cerebellum plays an important role in the differentiation, proliferation, and migration of neurons and gliacytes.

Cerebellar Cortex ; Fetus ; physiology ; Humans ; Immunohistochemistry ; Nerve Fibers ; Neurons ; cytology ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Nitric Oxide Synthase Type I