Objective To explore the changes of plasma brain natriuretic peptide(BNP) and N-terminal pro-B-type natriuretic peptide (NT-proBNP)of sepsis combined with myocardial injury in newborns.Methods According to neonatal sepsis treatment program,45 cases of sepsis newborns in NICU of the Second Hospital in Lanzhou University from Jul.2007 to Jun.2008 were collected.According to the myocardial injury diagnostic criteria,45 cases neonatal sepsis were divided into myocardial injury group(n=22) and non-myocardial injury group(n=23).Myocardial injury group was also divided into congenital heart disease group and non-congenital heart disease group accor-ding to echocardiography.At the same time,30 healthy newborns were collected as healthy control group.Every newborns were tested the level of plasma BNP,NT-proBNP,creatine kinase-MB(CK-MB) and cardiac-troponin I (cTnI).Results There were significant difference between myocardial injury group,non-myocardial injury group and healthy control group in the levels of plasma BNP,NT-proBNP,CK-MB and cTnI,those in congenital heart disease group were higher than those in non-myocardial injury group and the healthy control group(Pa0.05).Conclusions BNP and NT-proBNP can be early used to diagnose myocardial injury and heart failure of neonatal sepsis associated with CK-MB and cTnI.In NICU,infants with sepsis should normally test BNP and NT-proBNP in order to early diagnose myocardial injury of neonatal sepsis.

AIM To investigate the anti-neuroinflammation effects of 4-hydroxybenzyl aldehyde (4-HBAL) from Gastrodia elata Blume on acute cerebral ischemic injury in rats and its nechanism of action.METHODS The rat model of acute cerebral ischemic injury was induced by injecting arachidonic acid via intracarotid artery.Brain tissue samples were taken from the animals 3 h after the model of acute cerebral ischemic injury.Tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β) were detected in brain tissue to evaluate the effects of 4-HBAL in vivo.Lipopolysaccharid (LPS)-induced activation of BV-2 microglia cells model was used to explore the anti-neuroinflammation mechanism of 4-HBAL.RESULTS The experimental results showed that 4-HBAL had a significant protective effect on acute cerebral ischemic injury.It could significandy decrease the contents of tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β),and obviously inhibit the production of nitric oxide (NO),prostaglandin E2 (PGE2) and TNF-α in LPS-stimulated BV-2 cell,and increase the production of interleukin-10 (IL-10) and transforming growth factors-β (TGF-β) in BV-2 cell.CONCLUSION The mechanism of 4-HBAL may be related to the suppression of the excessive activation of microglia after cerebral ischemia and the promotion of the transformation of microglia into anti-inflammatory phenotype.

AIMTo study the influence of crocetin on cardiac hypertrophy induced by overloading pressure in rats.

METHODSThe model of cardiac hypertrophy was produced by overloading pressure in rats. The animals were divided into five groups: sham-operation group (0.5% CMC-Na, ig), model group (operation + 0.5% CMC-Na, ig), captopril group (operation + 50 mg x kg(-1), ig), crocetin I (100 mg x kg(-1), ig) and crocetin II (50 mg x kg(-1), ig). All animals were treated for 30 d by ig. Then, cardiac indexes were examined. The activity of ATPase and the hydroxyproline content in heart were assayed by colorimetric analysis. Matrix metalloproteinases (MMPs) activity was assayed by SDS-PAGE zymography.

RESULTSCompared with the model group, crocetin was found to significantly reduce the cardiac indexes and the content of hydroxyproline in heart, increase the activity of Na+ , K+ -ATPase, Ca2+, Mg2+ -ATPase and inhibit MMPs activity.

CONCLUSIONThe activity of MMPs may play a key role in the cardiac hypertrophy induced by overloading pressure, and proprably as a result of decreasing the activity of MMPs. Crocetin was shown to prevent remodeling of cardiac hypertrophy induced by overloading pressure.

Animals ; Ca(2+) Mg(2+)-ATPase ; metabolism ; Cardiomegaly ; enzymology ; metabolism ; Carotenoids ; pharmacology ; Hydroxyproline ; metabolism ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Myocardium ; enzymology ; Organ Size ; drug effects ; Rats ; Rats, Sprague-Dawley ; Sodium-Potassium-Exchanging ATPase ; metabolism

Background Spinal cord injury (SCI) is a serious neurological injury that often leads to permanent disabilities for the victims.The aim of this study was to determine the effects of glial-derived neurotrophic factor (GDNF) mediated by recombinant adeno-associated virus type 2 (rAAV2) alone or in combination with early rehabilitation training on SCI.Methods SCI was induced on the T8-9 segments of the spinal cord by laminectomy in adult male Sprague-Dawley rats.Then besides the sham operation group,the SCI rats were randomly divided into four groups:natural healing group,gene therapy group,rehabilitation training group,and combination therapy group (gene therapy in combination with rehabilitation training).Motor dysfunction,protein expression of GDNF,edema formation,and cell injury were examined 7,14,and 21 days after trauma.Results The topical application of rAAV-GDNF-GFP resulted in strong expression of GDNF,especially after the 14th day,and could protect the motor neuron ceils.Early rehabilitative treatment resulted in significantly improved motor function,reduced edema formation,and protected the cells from injury,especially after the 7th and 14th days,and increased the GDNF expression in the damaged area,which was most evident after Day 14.The combined application of GDNF and early rehabilitative treatment after SCI resulted in a significant reduction in spinal cord pathology and motor dysfunction after the 7th and 14th days.Conclusion These observations suggest that rAAV2 gene therapy in combination with rehabilitation therapy has potential clinical value for the treatment of SCI.

OBJECTIVENeonatal sepsis can cause multiple organ dysfunction syndrome, especially including myocardial injury and heart failure. In this study, the authors observed the changes and the levels of plasma brain natriuretic peptide and N-terminal pro-B-type natriuretic peptide in myocardial injury of neonatal sepsis at the different stages to search for the early diagnostic index of myocardial injury and heart failure in patients with neonatal septicemia.

METHODThe levels of plasma brain natriuretic peptide and N-terminal pro-B-type natriuretic peptide were determined in 96 newborns with neonatal septicemia according to the diagnosis and treatment program of neonatal septicemia in 2003. The 96 cases were divided into myocardial injury group and non-myocardial injury group. Every newborn was tested for the levels of plasma brain natriuretic peptide and N-terminal pro-B-type natriuretic peptide with enzyme-linked immunosorbent assay (ELISA) on the second day, fifth day and tenth day of septicemia and in the different gestational age infants. Meanwhile, the results were compared to creatine kinase isoenzyme and troponin I.

RESULTThe levels of plasma brain natriuretic peptide and N-terminal pro-B-type natriuretic peptide were significantly different between myocardial injury group and non-myocardial injury group at the fifth day (P<0.05), especially the levels of plasma N-terminal pro-B-type natriuretic peptide were significantly elevated at the early stage (on the second day) between the two groups (P<0.05). On the fifth day, the values of plasma N-terminal pro-B-type natriuretic peptide were (315.5 +/- 69.7) pmol/L in myocardial injury group, but the value of non-myocardial injury group was (179.3 +/- 27.5) pmol/L. On the fifth day, the results of plasma brain natriuretic peptide, N-terminal pro-B-type natriuretic peptide and troponin I were significantly different and had statistical significance between the myocardial injury group and non-myocardial injury group (P<0.05), while the results of creatine kinase isoenzyme had no statistically significant difference (P>0.05). The values of plasma brain natriuretic peptide were respectively (215.5 +/- 69.6) pmol/L and (119.3 +/- 37.4) pmol/L, While N-terminal pro-B-type natriuretic peptide were (315.5 +/- 69.7) pmol/L and (179.3 +/- 27.5) pmol/L in the two groups. The value of troponin I was (1.57 +/- 0.39) microg/L in the myocardial injury group and that in the non-myocardial injury group was (0.55 +/- 0.2) microg/L. The values of creatine kinase isoenzyme were (33.3 +/- 10.1) u/L in the myocardial injury group, but that of non-myocardial injury group was (17.4 +/- 8.5) u/L. In the different gestational age infants, the values of plasma brain natriuretic peptide and N-terminal pro-B-type natriuretic peptide of premature infants were the highest in the three groups. The values of plasma brain natriuretic peptide and N-terminal pro-B-type natriuretic peptide were (159.5 +/- 39.6) pmol/L and (238.5 +/- 49.7) pmol/L in premature infants.

CONCLUSIONThe levels of plasma brain natriuretic peptide and N-terminal pro-B-type natriuretic peptide evidently increased in myocardial injury of neonatal sepsis, especially in premature infants. The increase of plasma brain natriuretic peptide and N-terminal pro-B-type natriuretic peptide may be helpful in early diagnosis of the myocardial injury of neonatal sepsis associated with cTnI. N-terminal pro-B-type natriuretic peptide may become a useful index to diagnose the myocardial injury and should be widely used in the neonatal intensive care unit.

Female ; Heart Injuries ; blood ; diagnosis ; Humans ; Infant, Newborn ; Male ; Natriuretic Peptide, Brain ; blood ; Sepsis ; blood ; diagnosis

OBJECTIVETo investigate the inhibitory effects of OMT on TGF-β1-induced CFBs proliferation, and then explore the mechanism.

METHODThe experiment was randomly divided into 6 groups as following: control group (serum free DMEM), model group (20 μg x L(-1) TGF-β1), OMT low dose group (1.89 x 10(-4) mol x L(-1) + 20 μg x L(-1) TGF-β1), OMT medium dose group (3.78 x 10(-4) mol x L(-1) + 20 μg x L(-1) TGF-β1), OMT high dose group (7.56 x 10(-4) mol x L(-1) + 20 μg x L(-1) TGF-β1), SB203580 group (p38MAPK blocking agent, 1 x 10(-5) mol x L(-1) + 20 μg x L(-1) TGF-β1). Vimentin of CFBs was identified by immunocytochemical methods, α-SMA of myFBs as well. Inhibitory effects of OMT on CFBs proliferation was detected by the MTT assay. Picric acid Sirius red staining was analyzed collagen type I and collagen type III deposition. Western blot was determined the expression of p38MAPK, p-p38MAPK, collagen type I and collagen type III.

RESULTMTT results showed that OMT significantly inhibited CFBs proliferation induced by TGF-β1 (P < 0.01) α-SMA immunocytochemical experiments suggested that OMT could protect against the CFBs proliferation. OMT could significantly decrease the deposition of collagen type I and collagen type III by Western bloting and picric acid Sirius red staining. Western blot results showed that TGF-β1 enhanced p38MAPK phosphorylation, however OMT attenuated the phosphorylation of p38MAPK induced by TGF-β1 (P < 0.01).

CONCLUSIONOMT can inhibit the CFBs proliferation induced by TGF-β1, and its mechanism may be involved in inhibiting p38MAPK phosphorylation.

Alkaloids ; pharmacology ; Animals ; Cell Proliferation ; drug effects ; Collagen ; metabolism ; Down-Regulation ; Female ; Fibroblasts ; drug effects ; Heart ; drug effects ; In Vitro Techniques ; Male ; Phosphorylation ; Quinolizines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; antagonists & inhibitors ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; metabolism

OBJECTIVETo isolate and identify human periodontal ligament stem cells (PDLSC) by improved methods and assess the characteristics of PDLSC ex vivo.

METHODSThe periodontal ligament cells were obtained from the healthy impacted third molars and teeth extracted for orthodontic purposes and used to isolate PDLSC by limiting dilution assay. PDLSC were cultured and expanded in alpha-MEM supplemented with 10% FBS. Colony-forming assay, immunohistochemistry, flow cytometry, osteogenic and adipogenic induction were used to identify PDLSC.

RESULTSThe obtained cells had high colony-forming efficiency and were positive staining for vimentin and negative for pancytokeratin. Flow cytometry revealed that the isolated cells were positive for STRO-1 and CD146 antibodies and most were in the G0/G1 phase of cell cycle. Under specific conditions, they could differentiate to the osteoblast and adipocyte lineages in vitro.

CONCLUSIONLimiting dilution assay is an effective method to isolate PDLSC and the single-cell-derived colonies demonstrate the properties of stem cells in vitro.

Cell Differentiation ; Cell Separation ; Humans ; In Vitro Techniques ; Osteoblasts ; Periodontal Ligament ; Stem Cells

OBJECTIVETo investigate the expression of angiopoietin (Ang)-1 and Ang-2 in colorectal tumors and its relations to microvessel density (MVD) in tumor tissue.

METHODSAng-1, Ang-2 and factor VIII-related antigen were stained immunohistochemically in 91 cases of primary colorectal adenocarcinoma, 20 cases of colorectal adenoma and 24 cases of normal colorectal mucosal tissue, and MVD was also assayed in above tissue specimens.

RESULT(1) A significantly higher Ang-1 (7.07+/-2.00) was observed in normal tissue compared with 1.75 +/-1.98 in the adenoma and 1.40 +/- 1.22 in the adenocarcinoma (P<0.01). (2) Ang-2 protein positive rate in adenocarcinoma was significantly higher than that in normal tissue and adenoma (P<0.01). The expression of Ang-2 in adenocarcinoma was closely associated with poor differentiation and vessel invasion. (3) There were significant correlations between Ang-1 and Ang-2 (r=-0.338, P<0.01), Ang-1 and MVD (r=-0.388, P<0.01), Ang-2 and MVD (r=0.594, P<0.01) in the 135 cases.

CONCLUSIONThe overexpression of Ang-2 may play an important role in angiogenesis of colorectal adenocarcinoma. It can be regarded as an index for malignancy and prognosis in colorectal adenocarcinoma.

Adenocarcinoma ; blood supply ; metabolism ; Adult ; Aged ; Aged, 80 and over ; Angiopoietin-1 ; biosynthesis ; genetics ; Angiopoietin-2 ; biosynthesis ; genetics ; Biomarkers, Tumor ; Capillaries ; pathology ; Colorectal Neoplasms ; blood supply ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Prognosis ; von Willebrand Factor ; biosynthesis ; genetics

Objective To explore the problems of microsurgical treatment of multiple intracranial aneurysms with regard to the indications,surgical timing,operative approaches and the prevention of complications, and improve their diagnoses and treatments. Methods We retrospectively reviewed the data of 33 patients with multiple intracranial aneurysms admitted to our hospital from Jan.2000 to Dec.2008 and their clinical manifestations,imaging features and microsurgical treatments were summarized. Results In the 33 cases of multiple intracranial aneurysms,the male-to-female sex ratio was 1:1.75 and the average age at onset of symptoms was 56.52.Twenty-four patients showed good postoperative prognosis(Glasgow Outcome Scale IV-V) with 2 deaths.Conclusion Individual treatment should be adopted in the treatment of differently localized multiple intracranial aneurysms with different sizes and clinical classifications,and the utilization of temporary blockade and ultrasound during the operation can improve the therapeutic effects and reduce the postoperative complication rates.

Objective To develop a method for the determination of total cholesterol in serum by isotope dilution liquid chromatography tandem mass spectrometry (ID-LC/MS/MS).Methods Serum samples were supplemented by addition of [3,4-~(13)C_2]-cholesterol,hydrolyzed with alcoholic sodium hydroxide and oxidized into cholest-4-ene-3,6-dione by chromic acid.The oxidation products were analyzed by LC/MS/MS using atmospheric pressure chemical ionization (APCI) source and detection modes of multiple reaction monitoring (MRM) and single ion recording (SIR).Signals (peak areas) of the internal standard were corrected for the contributions of cholesterol and the signal ratios of cholesterol to internal standard for the calibrations were linearly regressed against cholesterol concentrations.The resulted regression equation was used for the calculation of serum cholesterol concentrations.Results The correlation coefficients between the peak area ratios and cholesterol concentrations were 0.999 9 and higher.Under MRM mode,the average within-run CV of the results obtained on 3 serum samples was 0.95% (ranged from 0.92% to 0.99%) and the total CVwas 0.86% (0.82% to 0.89%),and under SIR mode,the within-run CV was 0.64% (from 0.54% to 0.77%) and the total CVwas O.69% (0.62% to 0.81%),respectively. Results on certified reference materials (SRM 1951 a Level Ⅰ and Level Ⅱ;GBW 09145 and GBW 09147) showed an average bias of 0.23% (0.14% to 1.00%) under MRM mode,and 0.24% (0.07% to 1.27%) under SIR mode.Conclusions An ID-LC/MS/MS method for serum cholesterol has been developed.It is specific and precise and may be used as a candidate reference method.