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Background: The present study was designed to elucidate the relationship of engraftment efficiency of transplanted cells and Kupffer cell function in mice with acute on chronic liver injury and acute liver injury., Methods: The recipient dipeptidyl peptidase IV knockout (DPPIV(-/-)) mice were divided into two groups: (1) the acute on chronic liver injury group (CCl(4)/APAP group) that received CCl(4) (1 ml/kg) twice a week for 4 weeks following one dose of acetaminophen (APAP), 600 mg/kg; (2) the acute liver injury group (APAP-only group) that received a single dose of APAP at 600 mg/kg. DPPIV(+/+) hepatocytes were transplanted 24 h after APAP intoxication. Engraftment efficiency was evaluated at day 7 and day 14 after transplantation. The tumor necrosis factor-alpha (TNF-alpha) mRNA expression level of Kupffer cells immediately before cell transplantation was compared between the two groups before and after lipopolysaccharide (LPS, 100 ng/ml) stimulation., Results: The number of transplanted cells and clusters in each 100x microscopic field were higher in the CCl(4)/APAP group at both day 7 (21.5 +/- 6.3 versus 8.3 +/- 4.0, p < 0.001; 14.9 +/- 4.6 versus 6.6 +/- 3.4, p < 0.001, respectively) and day 14 (17.3 +/- 4.4 versus 10.2 +/- 3.3, p = 0.001; 12.6 +/- 3.2 versus 7.9 +/- 1.6, p = 0.004, respectively). After LPS stimulation, the expression level of TNF-alpha was lower (175.7 +/- 54.6 versus 465.6 +/- 64.2, p = 0.002), and the increment of TNF-alpha expression was also less significant in the CCl(4)/APAP group (1.5-fold versus 6.5-fold, p = 0.014)., Conclusions: Chronic liver injury desensitized Kupffer cells and reduced TNF-alpha expression, two results that correlated with the increased engraftment of transplanted cells.

Bone morphogenetic proteins (BMPs) are a class of morphogens that are critical regulators of the central nervous system (CNS), peripheral nervous system, and craniofacial development. Modulation of BMP signaling also appears to be an important component of the postnatal stem cell niche. However, describing a comprehensive model of BMP actions is complicated by their paradoxical effects in precursor cells, which include dorsal specification, promoting proliferation or mitotic arrest, cell survival or death, and neuronal or glial fate. In addition, in postmitotic neurons BMPs can promote dendritic growth, act as axonal chemorepellants, and stabilize synapses. Although many of these responses depend on interactions with other incoming signals, some reflect the recruitment of distinct BMP signal transduction pathways. In this review, we classify the diverse effects of BMPs on neural cells, focus on the known mechanisms that specify distinct responses, and discuss the remaining challenges in identifying the cellular basis of BMP pleiotropism. Addressing these issues may have importance for stem cell mobilization, differentiation, and cell integration/survival in reparative therapies.

Loss of genomic imprinting of insulin-like growth factor II (IGF2) is a hallmark of many human neoplasms. We attempted to correct this aberrant epigenotype by transferring nuclei from human tumor cells that showed loss of IGF2 imprinting into enucleated mouse and human fibroblasts that had maintained normal IGF2 imprinting. After nuclear transfer, the abnormal biallelic expression of IGF2 in tumor nuclei transiently converted to normal monoallelic imprinted expression in the reconstructed diploid cells. In tetraploid hybrid cells, however, normal IGF2 imprinting was permanently restored in the tumor genome. Inhibition of the synthesis of putative trans imprinting factors with cycloheximide led to loss of IGF2 imprinting in normal cultured fibroblasts, suggesting that normal cells produce proteins that act in trans to induce or maintain genomic imprinting. These data demonstrate that an abnormal tumor epigenotype can be corrected by in vitro reprogramming, and suggest that loss of imprinting is associated with the loss of activity of non-CTCF trans imprinting factor(s) that are either inactivated or mutated in tumors.

Objective: To explore the variations of early-phase insulin secretion in Type 2 diabetic patients in different stages., Methods: L-arginine stimulative test, fast blood glucose and body mass index (BMI) were evaluated in 40 nomal controls (NC) and 101 Type 2 diabetic patients. The diabetic patients were divided into three groups: newly diagnosed group (n = 35), effectively treated by sulfonylureas group (n = 32) , and secondary failure of sulfonylureas group (n = 34). The indexs of insulin resistance of homeostasis model assessment (HOMA-IR), beta-cell insulin secretion of homeostasis model assessment (HOMA-IS), and the acute insulin response (AIRARG) index were calculated. Some statistical comparisons were done among the 4 groups., Results: The indexs of HOMA-IR in each group of Type 2 diabetic patients were all higher than those in NC group (P < 0.01). The AIRARG indexs were obviously lower in Type 2 diabetic patients in different stages than those in NC group (P < 0.01), and the subsequence from the highest to the lowest among the groups of diabetic patients was: the newly diagnosed group, the effectively treated by sulfonylureas group, and the secondary failure of sulfonylureas group (P < 0.01). But there was no significant difference in indexs of HOMA-IS between the newly diagnosed group and the effectively treated by sulfonylureas group., Conclusion: There is severe insulin resistance in Type 2 diabetic patients in each stage. The variations of early-phase insulin secretion manifest a vary procedure of obvious deterioration by degrees from the newly diagnosed group to the secondary failure of sulfonylureas group in Type 2 diabetic patients.

Background: Neisseria meningitidis is one of the most significant bacterial infections in children and adolescents. As transplacental antibodies in the circulation gradually decline, the prevalence of meningococcal disease among young infants is high, and often presents an invasive clinical manifestation. The purpose of the study was to investigate the clinical spectrum of meningococcal infection in young infants., Methods: We retrospectively reviewed of the medical charts and analyzed the clinical characteristics and outcomes of 10 infants younger than 6 months old with meningococcal disease at the Chang Gung Children's Hospital from 1994 through 2004., Results: A total of 10 male infants with a mean onset age of 2.9 +/- 1.79 months old were enrolled. All patients presented initial symptoms of fever and decreased activity. Seizure attack was noted in six cases, and only three patients had purpuric or petechial rash. Laboratory findings reflected pyogenic infection including elevated C-reactive protein (159.1 +/- 108.8 mg/L), pleocytosis (791.11 +/- 660.83/microL), high protein levels (190.43 +/- 157.91 g/dl) and hypoglycorrhachia (28 +/- 20.89 mg/dl) in the cerebrospinal fluid. Seven cases presented meningitis; among those, N. meningitidis was isolated from blood in two cases. Three of the remaining patients had meningococcemia. Penicillin was the most common drug of choice; cephalosporin was the alternative. Prolonged antimicrobial therapy (range, 14 to 42 days) was prescribed in six patients complicated with subdural empyema. No deaths were documented. During long-term follow up, two patients developed mental retardation, and one of those two also had epilepsy. Both of them had lower birth body weight, altered initial conscious level, leukocytopenia and subdural empyema with encephalomalacia on brain images. One had insufficient therapy and another one was infected by a penicillin resistant strain., Conclusions: Clinicians should be aware of meningococcal infection in young infants because the initial presentations may be difficult to distinguish from viral syndrome, and may rapidly progress to clinical deterioration. Patients with subdural empyema required prolonged courses of antimicrobial therapy. Brain images confirmed the presence of encephalomalacia which increased the risk of permanent neurologic deficit.

Background/aims: BSEP, MRP2, and MDR3 are major hepatic canalicular transporters mediating bile secretion. Their expression in human liver during development has not been reported., Methods: Human liver samples from fetus at gestational age 14-20 weeks, adult livers and liver samples of infants with biliary atresia were tested for mRNA expression of BSEP, MDR3, MRP2, NTCP, FIC1, and FXR genes by using real-time RT-PCR. Immunohistochemical staining of BSEP, MDR3, and MRP2 were performed on fetal and adult livers., Results: All the genes tested were expressed at mid-gestational age. MDR3 and NTCP showed significant lower levels in fetal livers compared to adults. In patients with biliary atresia, all the genes tested showed higher mean expression levels than adults except for NTCP, but not statistically significant. The immunohistochemical staining of MRP2 in fetal liver was canalicular, BSEP showed both intracellular and canalicular staining, and MDR3 staining was faint, only occasional canalicular pattern could be seen., Conclusions: The major canalicular transporter genes are expressed at mid-gestational stage during human fetal development, but are different in expression level and targeting pattern, indicating differential regulation and maturation.

Recent studies suggest that IVF and assisted reproduction technologies (ART) may result in abnormal genomic imprinting, leading to an increased frequency of Angelman syndrome (AS) and Beckwith-Weidemann syndrome (BWS) in IVF children. To learn how ART might alter the epigenome, we examined morulas and blastocysts derived from C57BL/6J X M. spretus F1 mice conceived in vivo and in vitro and determined the allelic expression of four imprinted genes: Igf2, H19, Cdkn1c and Slc221L. IVF-derived mouse embryos that were cultured in human tubal fluid (HTF) (Quinn's advantage) media displayed a high frequency of aberrant H19 imprinting, whereas in vivo and IVF embryos showed normal maternal expression of Cdkn1c and normal biallelic expression of Igf2 and Slc221L. Embryonic stem (ES) cells derived from IVF blastocysts also showed abnormal Igf2/H19 imprinting. Allele-specific bisulphite PCR reveals abnormal DNA methylation at a CCCTC-binding factor (CTCF) site in the imprinting control region (ICR), as the normally unmethylated maternal allele acquired a paternal methylation pattern. Chromatin immunoprecipitation (ChIP) assays indicate an increase of lysine 4 methylation (dimethyl Lys4-H3) on the paternal chromatin and a gain in lysine 9 methylation (trimethyl Lys9-H3) on the maternal chromatin at the same CTCF-binding site. Our results indicate that de novo DNA methylation on the maternal allele and allele-specific acquisition of histone methylation lead to aberrant Igf2/H19 imprinting in IVF-derived ES cells. We suggest that ART, which includes IVF and various culture media, might cause imprinting errors that involve both aberrant DNA methylation and histone methylation at an epigenetic switch of the Igf2-H19 gene region.

Purpose: Pancreatitis-associated protein (PAP) and regenerating protein 1 alpha (Reg1A) are up-regulated during the pancreas regeneration. This study is to investigate the clinicopathologic denotation of their expression in hepatocellular carcinoma (HCC)., Experimental Design: PAP and REG1A mRNA levels were measured in 265 surgically removed unifocal primary HCCs using reverse transcription-PCR., Results: PAP and REG1A mRNAs were detected in 97 (36.6%) and 55 (20.8%) HCCs, respectively, including 46 with coexpression but in none of the 219 nontumorous livers. HCCs with PAP expression correlated with low-stage tumors without evidence of vascular invasion (P = 0.013) but the REG1A expression did not. By a combination analysis, HCCs with PAP expression alone showed the lowest frequency of p53 mutation (P < 0.036), the highest rates of grade 1 and low-stage tumors (P < 0.007 and P < 0.001, respectively), less frequent early tumor recurrence (P = 0.051), and hence a better 5-year survival (P = 0.044) than groups expressing PAP and REG1A, REG1A alone, and neither PAP or REG1A. Besides, PAP expressing HCCs had significantly frequent beta-catenin mutation, regardless of REG1A expression, P < 0.00001. In the subset of HCCs that has no mutations of p53 and beta-catenin but showed PAP expression, coexpression of REG1A and PAP was associated with more frequent vascular invasion than PAP expression alone (P < 0.005)., Conclusions: These data suggest that PAP expression designate a subset of low-grade, low-stage HCC with frequent beta-catenin mutation and hence more favorable prognosis, whereas further genetic or epigenetic alterations, such as p53 mutation and REG1A expression, lead to more advanced HCCs.

This study delineates the antiproliferative activities and in vivo efficacy of YC-1 [3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole] in human hepatocellular carcinoma cells. YC-1 inhibited the growth of HA22T and Hep3B cells in a concentration-dependent manner without significant cytotoxicity. YC-1 induced G(1) phase arrest in the cell cycle, as detected by an increase in the proportion of cells in the G(1) phase using FAC-Scan flow cytometric analysis. It was further shown that cGMP, p42/p44 mitogen-activated protein kinase, or AKT kinase-mediated signaling pathways did not contribute to the YC-1-induced effect. Of note, YC-1 induced a dramatic increase in the expression of cyclin-dependent kinase (CDK)-inhibitory protein, p21(CIP1/WAP1), and a modest increase in p27(KIP1). The association of p21(CIP1/WAP1) with CDK2 was markedly increased in cells responsive to YC-1. YC-1 did not modify the expression of cyclin D1, cyclin E, CDK2, or CDK4. In a corollary in vivo study, YC-1 induced dose-dependent inhibition of tumor growth in mice inoculated with HA22T cells. Immunohistochemical analysis revealed an inverse relationship between the staining of p21(CIP1/WAF) and the staining of Ki-67, a cell proliferation marker. Based on the results reported herein, we suggest that YC-1 induces cell cycle arrest and inhibits tumor growth both in vitro and in vivo via the up-regulation of p21(CIP1/WAP1) expression in HA22T cells. Because of this, YC-1 is a potential antitumor agent worthy of further investigation.

The large amount of absorbed dietary lipid after feeding a high-fat diet is mainly transported as triacylglycerol (TG)-rich lipoproteins (TRL) in the post-prandial blood and is subsequently distributed to peripheral tissues including adipose and muscle tissues. An in vivo and an in vitro study were conducted to investigate the possible role of post-prandial TRL after high fat feeding in the regulation of obese (ob) gene expression. Adult male Wistar rats were fasted for 48 h and re-fed either a fat-free/high-carbohydrate diet or a high-fat diet for 2, 4, or 8 h and plasma glucose, insulin, TG, and leptin as well as ob mRNA expression in epididymal fat pads were examined. Rats re-fed the high-fat diet had significantly higher plasma TG (p < 0.05) and lower plasma leptin and adipose ob mRNA (p < 0.05) than those fed the fat-free/high-carbohydrate diet; however, plasma glucose and insulin concentrations were not significantly different between the two groups. Plasma lipid analysis found large amount of TRL in rats fed with high-fat diet; however, only very small amount of the TRL was found in rats fed with fat-free/high-carbohydrate diet. We speculated that TRL might involve in regulation of ob gene expression. To further examine the regulation of TRL on ob mRNA expression, differentiated 3T3-L1 adipocytes were treated with TRL collected from rats fed 5 ml soybean oil by gastric intubations. TRL down-regulated ob mRNA not only in a dose and time dependent manner but also in the presence of insulin in 3T3-L1 adipocytes. These results suggest a possible role of TRL in the down-regulation of adipose ob mRNA expression and may account, at least in part, for the previous observations that short-term high fat feeding resulted in lower plasma leptin.

Objective: To prospectively investigate the relationship between high-sensitive C-reactive protein (hs-CRP) and Type 2 diabetic nephropathy., Methods: We assayed serum hs-CRP concentration in 55 normal controls, 66 Type 2 diabetic patients without diabetic nephropathy (DN) and 68 Type 2 diabetic patients with DN by ELISA. A 5-year prospective study was designed to investigate the changes of urinary albumin excretion (UAE), serum hs-CRP concentration, and kidney funciton in targeted intervention including blood glucose and blood pressure in 58 diabetic patients without DN at the baseline., Results: Baseline hs-CRP concentration was the highest in Type 2 diabetic patients with DN, followed by Type 2 diabetic patients without DN, and the lowest in normal controls. After 5-year intervention, hs-CRP concentration at the endline was significantly lower than that at the baseline in the patients in whom DN didn't occur at the endline (P < 0.05). But in the patients in whom DN occured at the endline, hs-CRP concentration was slightly lower than that at the baseline (P > 0.05). Both baseline and endline, the hs-CRP concentrations in the patients in whom DN occured at the endline were significantly higher than those without DN at the endline., Conclusion: Higher serum hs-CRP concentration in patients with Type 2 diabetes may be a risk factor that gives rise to DN. To some extent hs-CRP can predict the occurrence of DN in Type 2 diabetic patients.

Objective: To investigate the effects of rosiglitazone on serum leptin and insulin resistance (IR) in patients with Type 2 diabetes and to determine the correlation between IR and serum leptin., Methods: Thirty-nine Type 2 diabetic patients were given rosiglitazone (4 mg/d) for 12 weeks. The changes of height,weight, fasting plasm glucose (FPG), fasting serum insulin (FINS), serum leptin, hepatic and renal function and serum lipid were examined at the baseline and 12 weeks after the therapy. We used the HOMA model to calculate HOMA-IR as an index of insulin resistance. We also calculated the insulin sensitivity index (ISI). The correlation among the above detected markers was analyzed using multiple regression analysis and partial correlation analysis, respectively., Results: The serum leptin level, FINS, and HOMA-IR were higher (P < 0.01), but ISI was lower in the diabetic patients than those of the normal controls before the treatment. BMI, hepatic function, renal function, and serum lipid level did not significantly change, while FPG, FINS, and HOMA-IR were reduced significantly after the 12-week treatment (P < 0.01). ISI was obviously higher than that before the treatment (P < 0.01). Bivariate analysis showed that the serum leptin was positively correlated with FINS and HOMA-IR (r1 = 0.525, P < 0.01, r2 = 0.391, P < 0.05), but negatively correlated with ISI (r = -0.334, P < 0.05). Multiregressive analysis revealed that the most important factors affecting the leptin in Type 2 diabetes were sex, BMI, TG, FINS, and ISI (R2 = 0.358, P < 0.01)., Conclusion: There is close correlation between IR and higher serum leptin level in patients with Type 2 diabetes. Rosiglitazone can reduce FPG level and serum leptin and improve the insulin resistance in patients with Type 2 diabetes.

Hepatocyte transplantation has been proposed as an alternative for rescuing patients with acute hepatic failure. However, portal hemodynamic changes and issues of safety after hepatocyte transplantation in acute hepatic failure have not been systemically evaluated because of the lack of a suitable experimentation system. In this study, we created a novel spring-guidewire introducer needle to simplify the technique for long-term portal cannulation in F-344 rats. The portal cannula was capable of being used for blood sampling, infusion of hepatocytes, and measurement of portal hemodynamic changes. One week after portal cannulation, rats were injected with D-galactosamine (1.35 g/kg, i.p.) to induce hepatic failure. Hepatocytes (2 x 10(7)) were infused intraportally 24-26 h after induction of liver injury. Portal pressures were recorded for up to 60 min after hepatocyte transplantation. Intraportal infusion of 2 x 10(7) hepatocytes caused an instantaneous onset of portal hypertension. The magnitude of the rise in portal pressure was similar in both normal rats and rats with acute hepatic failure (33.0 +/- 7.1 vs. 37.7 +/- 0.5 mm Hg; p = 0.23). However, the resolution rate of portal hypertension was remarkably delayed in rats with acute hepatic failure, and the portal pressure was significantly higher than that in normal rats 60 min after hepatocyte transplantation (25.0 +/- 2.8 vs. 14.5 +/- 2.4 mm Hg; p = 0.007). In conclusion, we have established a simple new technique for long-term portal cannulation of rats. Our studies provide critical insights into the delayed resolution of portal hemodynamics after hepatocyte transplantation in subjects with acute hepatic failure., (2004 National Science Council, ROC and S. Karger AG, Basel)

Although viral replication and overwhelming immune responses are believed to contribute to the progression of severe acute respiratory syndrome (SARS), little is known about the temporal relationship between viral load, ribavirin, proinflammatory cytokines, and clinical progression. We report that ribavirin was not effective in reducing the SARS coronavirus load in 3 of 8 probable cases studied and that elevated levels of interleukin (IL)-6 and IL-8 subsequent to the peak viral load were found in 8 and 6 cases, respectively. The nadir lymphocyte count during lymphopenia, the peak level of lactate dehydrogenase, and the peak density of pulmonary infiltrates lag further behind the peak viral load by a median of 4, 5, and 3.5 days, respectively. These findings provide important information for therapeutic strategies to treat SARS.

Background: Reactivation of HBV replication is a clinically significant complication in HBV(+) patients receiving chemotherapy. We recently found that nearly half of the HBV reactivation in lymphoma patients occurred within 2 weeks of the first dose of chemotherapy. We hypothesized that mechanisms other than immunosuppression, such as direct stimulation of HBV replication by anticancer drugs, might be involved in this type of HBV reactivation., Materials and Methods: 2.2.15 cells, which secrete HBV particles constitutively, were used in the experiments. Real-time quantitative polymerase chain reaction was used to quantitate HBV DNA, and microparticle enzyme immunoassay to measure HBV surface antigen (HBsAg)., Results: HBV DNA secretion in culture medium was dose- dependently increased by doxorubicin, one of the most commonly used anticancer drugs for lmphoma. One-hour exposure of cells to 1 microM doxorubicin induced a 15.4+/-5.9-fold and a 3.05+/-0.09-fold increase of HBV DNA and HBsAg on the 4th culture day, respectively. Lamivudine suppressed the doxorubicin-induced increase of HBV DNA., Conclusion: Our data suggest that cytotoxic agents may stimulate the replication of HBVand thereby contribute to the reactivation of HBV during systemic chemotherapy. Importantly, this adverse effect of cytotoxic agents may be preventable by co-administration of lamivudine.

Objective: To prospectively clarify the relationship between angiotensin converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism and diabetic retinopathy (DR) in Type 2 diabetic patients., Methods: A 6-year prospectively study was designed to investigate the change of retina of 72 Type 2 diabetic patients without retinopathy . All patients suffered from diabetes mellitus for more than 5 years and matched well in age, body mass index (BMI), mean arterial pressure (MAP), and fasting blood glucose (FBG). Patients were classified into 3 groups according to their genotypes of ACE. ACE gene I/D polymorphism was identified by polymerase chain reaction. The patients were followed up for 6 years and their BMI, serum creatinine (Scr), MAP, HbA1c, and retina were checked once every 1 - 2 years., Results: Seven subjects (9.7%) were discontinued prematurely. At the end of the study, there were no significant differences in the clinical parameters such as BMI, MAP, FBG, HbA1c, and Scr among the 3 groups (P > 0.05), and also in DR incidence in Type 2 diabetic patients among the 3 groups (II 67.9%, ID 69.6%, DD 64.3%, respectively, P > 0. 05)., Conclusion: There is no association between ACE gene I/D polymorphism and the genesis and development of DR in Type 2 diabetic patients.

The severe acute respiratory syndrome-associated coronavirus (SARS-CoV) is thought to be transmitted primarily through dispersal of droplets, but little is known about the load of SARS-CoV in oral droplets. We examined oral specimens, including throat wash and saliva, and found large amounts of SARS-CoV RNA in both throat wash (9.58 x 10(2) to 5.93 x 10(6) copies/mL) and saliva (7.08 x 10(3) to 6.38 x 10(8) copies/mL) from all specimens of 17 consecutive probable SARS case-patients, supporting the possibility of transmission through oral droplets. Immunofluorescence study showed replication of SARS-CoV in the cells derived from throat wash, demonstrating the possibility of developing a convenient antigen detection assay. This finding, with the high detection rate a median of 4 days after disease onset and before the development of lung lesions in four patients, suggests that throat wash and saliva should be included in sample collection guidelines for SARS diagnosis.

Objective: To investigate the change of serum TNF-alpha level in type 2 diabetes mellitus after treating with rosiglitazone, and the relationship between TNF-alpha and insulin resistance., Methods: The level of serum TNF-alpha in 27 patients was tested by radioimmunoassay before and after treating with rosiglitazone. Free insulin, free blood sugar, TG, HDL-C and weight were measured, and HOMA-IR was calculated., Results: The level of serum TNF-alpha in the patients was higher than that in the controls (P < 0.005) and rosiglitazone could decrease the level of serum TNF-alpha (P < 0.001). The level of serum TNF-alpha was positively related to serum TNF-alpha, TG, FBS and ln (HOMA-IR), and it was negatively related to HDL-C (r = -0.508, P < 0.05)., Conclusion: Rosiglitazone can decrease the level of serum TNF-alpha significantly and improve insulin resistance.

Bone morphogenetic proteins (BMPs), BMP receptors (BMPRs), and endogenous BMP antagonists have been found to be critically important for the development of the central nervous system (CNS) and peripheral organs in mammals. There is also increasing evidence that this system has significant activity in the adult CNS. Accordingly, we studied the regional distribution of endogenous BMP ligand proteins, receptors, and antagonists during aging and after lesion of the midbrain dopamine pathways produced by 6-hydroxydopamine (6-OHDA). We found that there were only small changes in the levels of these molecules as a function of age. Interestingly, levels of BMP 7 and noggin, a BMP antagonist, were uniquely elevated in substantia nigra. Moreover, after lesions of the midbrain dopamine system by 6-hydroxydopamine, there was a marked reduction in levels of all BMP ligands, receptors and antagonists bilaterally in both substantia nigra and hippocampus. There were also differential changes in BMP ligands, receptors, and antagonists in the cortex and striatum after such lesions. Taken together, our results indicate significant expression of BMP-related molecules in the adult and aging brain, and suggest a dynamic and differential regulation of these molecules after perturbations.

Objective: To assess the relationship between serum leptin and insulin resistance (IR) in obesity, and to investigate the effects of sibutramine on obesity, serum leptin and IR., Methods: Seventy obese subjects [body mass index (BMI) > or =25 kg/m2] were randomly divided into 2 groups: group B (sibutramine 10 mg/day) and group C (a placebo tablet/day). Both had been treated for 12 weeks. Another 30 healthy adults served as the normal control (group A: BMI < 23 kg/m2). Their height, body weight, waist and hip circumference, fasting plasms glucose (FPG), fasting plasma insulin (FINS), and serum leptin were examined at the baseline and 12 weeks after the therapy. Insulin senstivity index (ISI) was calculated [ISI = 1/(FPG x FINS)]. Multiple linear regression analysis and partial correlation were performed on serum leptin., Results: The body weight, BMI, waist and hip circumference decreased significantly after the 12 week-treatment with sibutramine in group B (P < 0.01), but those indexes did not change after the treatment with placebo in group C (P > 0.05). The levels of leptin and FINS were higher (P < 0.01), but ISI was lower (P < 0.01) both in group B and C compared with those in group A at the baseline. The levels of serum leptin and FINS decreased (P < 0.01), and ISI increased significantly (P < 0.05) after the treatment with sibutramine in group B, while those indexes did not change after the treatment with placebo in group C. The most important factors to influence serum leptin level were listed as follows: sex > BMI > FINS > ISI (R2 = 0.661, F = 12.662, P < 0.01). The lep- tin was positively correlated with FINS (r = 0.597, P < 0.01) , but negatively correlated with ISI (r = -0.468, P < 0.01 ) after eliminating the effects of sex and BMI. Conclusion Leptin resistance and insulin resistance exist in obesity, and serum leptin is associated with IR. Treatment with sibutramine significantly reduces the body weight and leptin, increases insulin senstivity, and improves IR.

Objective: To prospectively clarify the relationship between angiotensin converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism and diabetic nephropathy (DN) in Type 2 diabetic patients., Methods: We examined 66 Type 2 diabetic patients with normal buminure. All patients suffered from diabetes mellitus for more than 5 years and matched well in age, body mass index (BMI), mean arterial pressure (MAP), fasting blood sugar (FBS), and urinary excretion of albumin (UAE). Patients were classified into 3 groups according to genotypes of ACE. ACE gene I/D polymorphism was identified by polymerase chain reaction (PCR). The patients were followed up for 5 years and their UAE, serum creatinine (Scr), and HbAlc were checked once a year., Results: Eight subjects (12%) were discontinued prematurely. At the end of the study, there were no significant differences in the clinical parameters such as BMI, MAP, FBS, HbA1c, UAE, and Scr among the 3 groups (P > 0.05), and also in DN incidence in Type 2 diabetic patients among the 3 groups (II 45.8%, ID 52.3%, and DD 46.1%, respectively, P > 0.05 ). The increased degree of UAE among the 3 groups was similar at the end-point of the study (P > 0.05)., Conclusion: The DD genotype of ACE gene may not be a clinically useful genetic marker for predicting the genesis and development of DN in Type 2 diabetic patients in Chinese. There is no association between ACE gene I/D polymorphism and the genesis and development of DN in Type 2 diabetic patients.

Objectives: To investigate the level of AIDS knowledge among people concerned in Nanjing city in order to provide scientific evidence and constructive suggestions for the government to formulate relevant policies for AIDS control., Methods: Three sets of questionnaires on AIDS knowledge were designed, the scores calculated, and the results evaluated., Results: Of the 2,500 questionnaires issued to 4 different groups of people, 2,436 were collected back with effective answers, 991 from medical and health-related workers with the mean score of 58, 473 from college students with the mean score of 39.9, 524 from common city residents with the mean score of 42.3, and 448 from those working in high risk environment with the mean score of 47., Conclusions: The level of AIDS knowledge among people concerned in Nanjing city was far below the requirement of the nation, especially among medical and health-related workers. Efforts must be made to raise the level of AIDS knowledge of people concerned so as to enhance the prevention and treatment of the disease.

Expression of DNase II in macrophages is potentially crucially important in the removal of unwanted DNA. We have previously shown that DNase II expression is up-regulated at the transcriptional level during the phorbol 12-myristate-13-acetate (PMA)-induced differentiation of HL-60 and THP-1 cells. In this study, we investigated the cis-regulatory elements and transcription factors involved in this process in HL-60 cells. cis-Regulatory elements in the DNase II promoter were located by 5' deletion and site-directed mutagenesis of promoter-luciferase constructs and transient transfection of HL-60 cells. Furthermore, the binding proteins were identified by electrophoretic mobility shift assay (EMSA) in the presence of specific antibodies. In the DNase II promoter, 249 base pairs upstream of the transcription start site were essential for maximal promoter activity in both untreated and PMA-treated HL-60 cells and, within this region, three Sp1 and Sp3 binding sites were identified as essential for transcriptional regulation and PMA induction. Western blot analysis showed that PMA treatment resulted in increased levels of Sp1 and Sp3 proteins. Furthermore, cotransfection analysis in Drosophila SL2 cells showed that Sp1 was more potent than Sp3 in activating the DNase II promoter. We therefore conclude that Sp1 and/or Sp3 are involved in the up-regulation of DNase II expression during the differentiation of HL-60 cells.

Background and Purpose: We and others have previously reported that bone morphogenetic protein-7 (BMP-7), given before middle cerebral artery occlusion (MCAO), reduces ischemic injury in brain. Recent studies have indicated that receptors for BMP are upregulated after brain ischemia. It is possible that this upregulation may facilitate endogenous neurorepair in the ischemic brain. The purpose of this study was to determine the neuroregenerative effects of BMP-7 given parenterally after ischemia/reperfusion injury., Methods: Adult Sprague-Dawley rats were anesthetized with chloral hydrate. The middle cerebral artery was transiently occluded by a filament inserted through the right internal carotid artery. The filament was removed after 60-minute ischemia to allow reperfusion. Some animals were killed 24 hours after MCAO to examine BMP-7 mRNA expression. Other animals received a single dose of intravenous BMP-7 or vehicle at 24 hours after MCAO and were used for subsequent behavioral studies and BMP-7 immunostaining., Results: BMP-7 mRNA was upregulated 24 hours after MCAO in untreated animals. BMP-7 immunoreactivity was dose-dependently increased on the ischemic side of the hippocampus/dentate on day 6 after MCAO in animals receiving intravenous injection of BMP-7. Animals receiving BMP-7 also showed a decrease in body asymmetry from day 7 to day 14 and an increase in locomotor activity on day 14 after MCAO., Conclusions: Our data indicate that BMP-7, given parenterally after stroke, can pass through the blood-brain barrier on the ischemic side and induce behavioral recovery in stroke animals at longer testing times.

Several recent studies have suggested that intracellular deoxyribonuclease II (DNase II) is responsible for the degradation of DNA from apoptotic cells that are engulfed by macrophages. In this study, we studied DNase II expression during the phorbol 12-myristate-13-acetate (PMA)-induced differentiation of HL-60 and THP-1 cells. Basal levels of DNase II mRNA and protein were low, with expression being up-regulated approximately 15- and 7-fold, respectively, in HL-60 and THP-1 cells 72 h after PMA treatment. Nuclear run-on and luciferase reporter assays showed that transcription of DNase II gene was increased in PMA-treated cells. Together, these results demonstrate that DNase II gene transcription is increased during myelomonocytic differentiation, resulting in increased levels of mRNA and protein. This increase in DNase II levels in differentiated HL-60 and THP-1 cells suggests that it may play an important role in macrophages.