The present study was designed to investigate the role of protein kinase A (PKA) and phospholipase A(2) (PLA(2)) in the stimulating effect of adenosine on the basolateral 50 pS K(+) channels in the thick ascending limb (TAL) of the rat kidney. Under the anatomic microscope, the TAL was dissected. The current of 50 pS K(+) channels were recorded by patch clamp technology. The protein expression of phosphorylated PKA and phosphorylated PLA(2) were examined by Western blot. The results showed that cyclohexyladenosine (CHA), an analog of adenosine, increased the 50 pS K(+) channel activity (P < 0.05). In the presence of H8, an antagonist of PKA, CHA did not affect the 50 pS K(+) channel activity. In the presence of AACOCF3 (an antagonist of PLA(2)), CHA did not further increase the 50 pS K(+) channel activity. CHA increased phosphorylation level of PKA, whereas inhibited phosphorylation of PLA(2) in the TAL of the rat kidney (P < 0.01). Furthermore, after blocking the PLA(2) with AACOCF3, CHA still increased the expression of phosphorylated PKA. On the contrary, CHA did not obviously change the expression of phosphorylated PLA(2) after H8 pretreatment. The results suggest that the stimulation of basolateral 50 pS K(+) channels by CHA is mediated by the activation of PKA followed by the inhibition of PLA(2) in the TAL of the rat kidney.
Adenosine ; analogs & derivatives ; pharmacology ; Animals ; Arachidonic Acids ; pharmacology ; Cyclic AMP-Dependent Protein Kinases ; metabolism ; Kidney ; drug effects ; metabolism ; Patch-Clamp Techniques ; Phospholipases A2 ; metabolism ; Potassium Channels ; metabolism ; Rats ; Signal Transduction

0.05),but a significant difference at weeks 4,8,and 12 between two groups(P

OBJECTIVETo study the expression of nNOS and ultrastructural changes in the penile tissue of rats with prolactinoma-induced erectile dysfunction (ED).

METHODSWe established the model of prolactinoma in 20 male Westar rats by peritoneal injection of diethylstilbestrol (DES) and treated the control rats with normal saline (n = 10) or sterilized arachis oil (n = 10). After 8 weeks, we performed the apomorphine test and measured the weight of the pituitary gland and the levels of serum prolactin (PRL) and testosterone (T) to confirm the successful construction of the prolactinoma-induced ED model. Then we determined the expression of nNOS in the penile tissue by immunohistochemistry and examined the ultrastructural changes of the penile cavernosum under the transmission electron microscope.

RESULTSThe prolactinoma-induced ED model was successfully established in 15 rats. The weight of the pituitary gland was significantly increased in the rats treated with DES as compared with the normal saline and sterilized arachis oil controls ([46.7 ± 15.5] vs [11.7 ± 2.4] and [12.4 ± 2.3] mg, both P < 0.05). The level of serum PRL was markedly higher while that of T remarkably lower in the former than in the latter two groups ([1,744.9 ± 304.5] vs [11.5 ± 2.4] and [10.6 ± 1.9] ng/ml, both P < 0.0l; [1.54 ± 0.46] vs [3.11 ± 1.08] and [3.04 ± 1.11] ng/ml, both P < 0.05). The rate of penile erection was significantly reduced in the prolactinoma-induced ED model rats in comparison with the normal saline and arachis oil controls (16.7% vs 100% and 87.5%, both P < 0.05), and so was the expression of nNOS in the penile tissue (0.024 ± 0.011 vs 0.066 ± 0.019 and 0.058 ± 0.021, both P < 0.05). Transmission electron microscopy manifested significant ultrastructural changes in the endothelial and smooth muscle cells of the cavernous tissue in the prolactinoma-induced ED models.

CONCLUSIONThe ultrastructural changes of the penile cavernous tissue and the reduced expression of nNOS in penile tissue may be the most important mechanisms of prolactinoma-induced ED in rats.

Animals ; Apomorphine ; Carcinogens ; Diethylstilbestrol ; Erectile Dysfunction ; etiology ; Humans ; Male ; Myocytes, Smooth Muscle ; ultrastructure ; Nitric Oxide Synthase Type I ; metabolism ; Organ Size ; Penile Erection ; Penis ; enzymology ; ultrastructure ; Pituitary Neoplasms ; chemically induced ; complications ; Prolactin ; blood ; Prolactinoma ; chemically induced ; complications ; Rats ; Rats, Wistar ; Testosterone ; blood

BACKGROUND: The tooth ash can be used as a scaffold for bone tissue growth and provide calcium and phosphorus components during bone regeneration. Platelet-rich plasma can promote the soft and hard tissue regeneration. However, either of them has its shortcomings.OBJECTIVE: To investigate the effect of platelet-rich plasma combined with tooth ash in repairing skull bone defects. METHODS: Nine healthy rabbits were selected to make bone defects on both sides of the skull. Rabbit platelet-rich plasma combined with tooth ash was implanted into the skull defect on the left side as experimental group, while rabbit platelet-rich plasma was implanted into the skull defect on the right side as control group. Skull samples were taken out at 4, 6, 8 weeks after implantation for soft X-ray detection, hematoxylin-eosin staining and modified Gomori staining. RESULTS AND CONCLUSION: (1) Soft X-ray: The trabecular bone area of the experimental group was larger than that of the control group at 6 and 8 weeks after implantation (P < 0.05 or P < 0.01). (2) Hematoxylin-eosin staining: With the duration of implantation, newly formed fibers with bone structure gradually reduced at the defect sites in both groups, and there was a orderly layered arrangement in the bone structure and increased calcification. Compared with the control group, relatively higher new bone maturity, better bone trabecular arrangement and more osteoblasts were observed in the experimental group. (3) Modified Gomori staining: With the prolongation of implantation time, the new bone became mature gradually in the two groups, and the bone maturity in the experimental group was higher than that in the control group. To conclude, platelet-rich fibrin combined with tooth ash is better to promote bone defect repair.

BACKGROUNDAngioplasty in the unprotected left main coronary artery (LMCA) has been controversial. This study aims to evaluate the safety and clinical effectiveness of stenting, including bare metal stent and drug eluting stent (DES), for treatment of unprotected LMCA disease.

METHODSBetween September 1997 and December 2005, a total of 297 consecutive patients underwent percutanous coronary intervention (PCI) on LMCA lesions in our hospital. Their in-hospital data and clinical follow-up outcomes were analyzed and those in pre-DES "era" (group I, from September 1997 to December 2002) were compared with those in DES "era" (group II, from January 2003 to December 2004. Patients in 2005 for the time of follow-up less than one year were not included in this group).

RESULTSAltogether 368 coronary stents were successfully deployed in 295 patients. Stents failed to be implanted after balloon predilation in two patients, who received coronary artery bypass graft (CABG) successfully. Bifurcation techniques for distal LMCA executed in 206 patients (69.4%, 206/297), included crossover stenting in 156 (75.7%), T stenting in 4 (1.9%), provisional T stenting in 28 (13.6%), kissing stenting in 5 (2.4%) and stent crushing in 13 (6.3%) patients. During their hospital stay, 5 (1.7%) patients died after PCI procedure, of which 4 died from cardiac origin and one of renal failure. The total in-hospital major adverse cardiac events (MACE) were 2.0% (6/297). In the follow-up period, 19 patients (6.5%) died [15 (5.1%) of cardiac death and 4 of non-fatal myocardial infarction (MI)]. Besides, 2 (0.7%) developed subacute thrombosis (SAT) and 16 (5.4%) performed target lesion revascularization (TLR). The total follow-up MACE was 14.5% (43/297). Further analysis also showed that, compared with patients in group I, those in group II apparently had more multi-vessel involvement (14.7% vs 81.9%, P < 0.001), and more bifurcation lesions (32.4% vs 72.2%, P < 0.001). After PCI, in-hospital MACE of group II was significantly lower than that in group I (1.1% vs 9.4%, P < 0.05). And the incidences of MACE, TLR and angiographic restenosis in group II were all significantly lower than those in group I (all P < 0.05) after one year follow-up.

CONCLUSIONSAs new PCI strategies and intervention devices such as DES are developed, coronary stenting, which might have brought better in-hospital and long-term outcomes than CABG, are proved to be technically successful and can be safely applied for the treatment of LMCA lesions in the experienced center for coronary intervention.

Adult ; Aged ; Aged, 80 and over ; Angioplasty, Balloon, Coronary ; Coronary Angiography ; Coronary Disease ; therapy ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Stents

OBJECTIVETo find out association mapping of loci related to schizophrenia on chromosome 1 with microsatellite markers in DNA pooling samples from schizophrenic cases and normal controls in Shandong peninsula.

METHODSA total of 31 microsatellite markers on chromosome 1 spaced at approximately 10 cM were scanned to two separated DNA pooling samples consisting of 119 schizophrenic cases and 119 normal controls respectively. Statistic analysis was performed by Chi-square test method to compare the difference between the ratio of each allele between the two pooling samples.

RESULTSSignificant statistic difference was found at D1S2878 between cases and controls, and P< 0.01 at this loci.

CONCLUSIOND1S2878 locus on chromosome 1 associates with schizophrenia in Shandong peninsula. Fine mapping and searching for candidate genes are warranted in this region.

Adult ; Case-Control Studies ; China ; Chromosome Mapping ; Chromosomes, Human, Pair 1 ; genetics ; DNA ; genetics ; Female ; Genomics ; Humans ; Male ; Microsatellite Repeats ; genetics ; Schizophrenia ; genetics ; pathology

OBJECTIVETo investigate the chronic efficacy of low-dose hydrochlorothiazide (HCTZ) in patients with mild-to-moderate hypertension.

METHODSAfter a 2-weeks placebo run-in period, 232 patients with mild or moderate hypertension were recruited and received HCTZ (12.5 mg once daily) therapy for one year. Patient compliance and blood pressure were monitored and serum BUN, Cr, glucose, electrolytes, and lipids were measured before, 6 weeks and 1 year after treatment.

RESULTS(1) Reduction of SBP, DBP and MAP were more significantly at 1 year [(10.45 +/- 17.28) mm Hg, (8.45 +/- 11.06) mm Hg, (9.12 +/- 10.88) mm Hg] than that at 6 weeks post therapy [(6.01 +/- 16.05) mm Hg, (2.90 +/- 10.33) mm Hg, (3.94 +/- 10.68) mm Hg, all P < 0.05]. Blood pressure were reduced to normal in 35.1% patients at 1 year and in 20.3% patients at 6 weeks (P < 0.05). (2) No patient developed diabetes mellitus or hypokalemia during therapy while the serum uric acid at 1 year post therapy was significantly higher than that at before therapy (P < 0.05).

CONCLUSIONThe study indicates that low dose HCTZ is an effective and safe antihypertensive agent for patients with mild-to-moderate hypertension and uric acid changes during therapy need to be monitored.

Adult ; Aged ; Antihypertensive Agents ; administration & dosage ; Blood Pressure ; Female ; Humans ; Hydrochlorothiazide ; administration & dosage ; Hypertension ; drug therapy ; physiopathology ; Male ; Middle Aged ; Treatment Outcome

OBJECTIVESevere newborn hypoxic-ischemic encephalopathy (HIE) has a very high rate of disability and no effective treatment is available. The present study aimed to preliminarily evaluate the effects of human neural stem cell transplantation in treatment of severe neonatal HIE.

METHODSThe patient was a 75-day old male infant with sequelae of severe HIE who had highly delayed development of intelligence and movement and myotonia. MRI showed multiple cerebromalacia and encephalatrophy. Cells obtained from the forebrain of an 11-week old fetus were cultured and amplified for 15 days. And then the human fetal neural stem cells were injected into cerebral ventricle of this infant.

RESULTSTwenty eight days after transplantation, remarkable improvement occurred not only in his myotonia but also in his intelligence and movement, which became similar to those of the normal infants of the same age. Positron emission tomography (PET) showed significantly increased radioactivity at temporal and occipital lobes which suggested that the cellular metabolism had increased greatly.

CONCLUSIONThe short-term effect of NSCs transplantation on the infant with severe HIE sequelae was significant. PET suggested that the implanted NSCs survived. Many more studies are needed to evaluate long-term effects of NSC transplantation in treatment of HIE.

Asphyxia Neonatorum ; complications ; Brain ; pathology ; physiopathology ; Female ; Humans ; Hypoxia-Ischemia, Brain ; etiology ; pathology ; physiopathology ; therapy ; Infant ; Infant, Newborn ; Injections, Intraventricular ; Multipotent Stem Cells ; transplantation ; Neurons ; Positron-Emission Tomography ; Prognosis ; Stem Cell Transplantation ; methods ; Time Factors ; Treatment Outcome

OBJECTIVENeonatal hypoxic-ischemic encephalopathy (HIE) harms the lives and health of newborn infants and children severely. Given the absence of effective therapies for HIE, it is important to derive new strategies. Neural stem cells (NSCs) have great potential as a therapeutic tool for the repair of a number of central nervous system disorders that involve cell loss. This study was designed to transplant the neural stem cells derived from human fetal brain (hNSCs) into cerebral ventricle of neonatal rat following hypoxic-ischemic injury and to investigate their survival, migration and differentiation in rat brain.

METHODSCells obtained from the forebrain of a 12-week old fetus were cultured in the presence of epidermal growth factor, basic fibroblast growth factor and leukemia inhibitory factor for 11 days. Animal models were built in 7-day-postnatal Wistar rats, 3-days after hypoxia-ischemia (HI), 5 microl suspension containing 5.0 x 10(5) hNSCs was injected into the left cerebral ventricle of each HIE rat by using stereotactic instrument. No immunosuppression therapy was given to the animals. At 1, 2, 4 weeks and 3 months after transplantation, the rats were sacrificed and brain tissues were harvested and were then examined by H-E staining and immunohistochemical analysis.

RESULTSImplanted cells expressing human nuclear protein (hNP) migrated form the subventricular zone (SVZ) along corpus callosum to the damaged areas, especially to the injured side of cortex and hippocampus. In different areas, the implanted hNSCs differentiated into different cell types which were similar to the host cells. The 85% implanted cells in cortex consisted of hNuc-NF or hNuc-Tublin double positive cells, while in the migratory way, 60% implanted cells differentiated into hNuc-GFAP double positive cells. Compared with the 1-week time point, an increased number of hNP-positive cells were observed at 2-weeks, but the number of these cells greatly decreased at 4-weeks and 3 months.

CONCLUSIONThe implanted hNSCs could extensively survive, migrate in the brain of neonatal rat with HIE and could differentiate into neurons and astrocytes in a regionally specific manner.

Animals ; Animals, Newborn ; Brain ; pathology ; Carotid Artery, Common ; surgery ; Cell Differentiation ; Cell Movement ; Disease Models, Animal ; Fetal Stem Cells ; transplantation ; Humans ; Hypoxia ; complications ; physiopathology ; Hypoxia-Ischemia, Brain ; pathology ; physiopathology ; therapy ; Immunohistochemistry ; Injections, Intraventricular ; methods ; Ligation ; methods ; Neurons ; Nuclear Proteins ; metabolism ; Rats ; Rats, Sprague-Dawley ; Stem Cell Transplantation ; methods ; Survival Analysis ; Time Factors

OBJECTIVETo detect lymphangiogenesis by labeling the lymphatic endothelial marker, lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1), and study the prognostic relevance of lymphangiogenesis in laryngeal squamous carcinoma.

METHODSClinical files and specimens of 78 patients with histologically diagnosed laryngeal carcinoma were stained with LYVE-1 as a specific lymphatic endothelial marker. The lymphatic vessel density (LVD) was measured, and the correlation between LVD and clinicopathological features of the tumor cases was analyzed.

RESULTSThe mean LVD in laryngeal carcinoma (13.24 ± 5.09) was significantly higher than that in adult laryngeal papilloma (5.54 ± 3.15) and squamous dysplasia (6.76 ± 4.45, P < 0.05). The LVD of poorly differentiated tumors (15.74 ± 5.24) was significantly higher than that in the moderately differentiated tumors (13.84 ± 6.20), and the LVD in the moderately differentiated tumors was significantly higher than that in the well-differentiated tumors (11.68 ± 6.34). The LVD in stage 0 to stage II group (10.66 ± 5.70) was significantly lower than that in the stage III to IV group (17.01 ± 6.35). The lymph node metastasis group (17.25 ± 7.37) was significantly higher than non-lymph node metastasis group (8.60 ± 5.23, P < 0.05). There was no significant association between LVD and age, sex, primary site and distant metastasis. The overall survival in the patients with a LVD higher than the mean value was 33.5 month, and that of cases with a LVD lower than the mean value was 81.6 month (P < 0.05). The multivariate survival analysis showed that the clinical stage and LVD were independent prognostic factors of laryngeal cancer.

CONCLUSIONSThe LYVE-1 staining histochemistry demonstrates that the lymphangiogenesis occurrs mainly at the edge of the tumors, and lymphangiogenesis plays an important role in the carcinogenesis, cancer progression and lymph node metastasis in laryngeal cancer. LVD may be an independent indicator of poor prognosis of laryngeal cancer.

Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Female ; Follow-Up Studies ; Humans ; Immunohistochemistry ; Laryngeal Neoplasms ; metabolism ; pathology ; Lymphangiogenesis ; Lymphatic Metastasis ; Lymphatic Vessels ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Papilloma ; metabolism ; pathology ; Precancerous Conditions ; metabolism ; pathology ; Survival Rate ; Vesicular Transport Proteins ; metabolism