Adolescent ; Adult ; Angiography, Digital Subtraction ; Child ; Decompression, Surgical ; methods ; Female ; Humans ; Magnetic Resonance Angiography ; Male ; Middle Aged ; Optic Nerve Injuries ; diagnostic imaging ; pathology ; Young Adult

Objective To study the transfection efficiency, protein expression, and effect of adenovirus-mediated transfection on microvascular endothelial cells transfected by angiopoietin-related protein 2(Ad. ARP2)gene. Methods Mice coronary microvascular endothelial cells(CMECs) were isolated, cultured and transferred by Ad-ARP2. The transfection efficiency and cellular toxicity of adenovirus vector to CMECs were detected by immunofluorescence staining. Expression of Ad. ARP2 in CMECs and the secreted materials in culture medium were measured by Western blot and ELISA and compared among groups of Ad. ARP2, Ad. null, and PBS control. Vascular endothelial cells were incubated with conditional medium containing secreted ARP2, and effects on cells sprouting were observed in matrigel. Results Adenovirus-transfected CMECs showed a very high efficiency. When multiplicity of infection (MOD was 200, the transfection efficiency was 93. 5% ,and no harmful effect on CMECs growth was found. When CMECs were transfected with Ad. ARP2, there was a high ARP2 expression, which was significantly different from that with Ad. Null or PBS. The conditional medium containing ARP2 had an excellent ability to stimulate sprout of CMECs which phenomenon could not be seen in the control groups. Conclusions Adenovirus vector can be transferred into CMECs efficiently and safely. Ad. ARP2 gene transfection allows a high transient expression, and the expression products can stimulate the sprout of microvascular endothelial cells in vitro very well.

OBJECTIVETo reveal the quality variation of polysaccharides, triterpenoids and proteins in spores and fruiting bodies of Ganoderma lucidum from producing areas, different varieties, harvesting parts and periods, and wall-breaking treatments.

METHODSpores and fruiting bodies from varieties of Longzhi No. 1 and Hunong No. 1 were collected as test samples, together with wall-broken spores sold in domestic main producing areas. The anthrone-sulfuric acid colorimetric method was used to determine the content of total polysaccharides. The vanillin-glacial acetic acid-perchloric acid colorimetric method was used to determine the content of total triterpenoids. The Lowry method was used to determine the content of total proteins.

RESULTThe content ranges of total polysaccharides, total triterpenoids, and total proteins from 6 domestic main producing areas were 0.40% - 2.25%, 1.36%-3.15% and 0.74% -1.91% respectively. The content ranges of total polysaccharides, triterpenoids, and proteins in the fruiting bodies from 2 varieties cultured in Zhejiang were 0.25% -1.42%, 0.44% -1.42% and 1.82% -3.67% respectively. In addition, the ranges of samples from wall-unbroken spores were 0.41% - 0.91%, 0.09% - 0.12%, 0.78% - 0.90% respectively and wall-broken spores are 1.03% - 2.25%, 1.89% - 3.15%, 0.96% - 1.04% respectively.

CONCLUSIONThere are significant differences in the contents of main chemical ingredients of wall-broken G. lucidum spores saled in the markets. The samples from Zhejiang contain high content of total polysaccharides and triterpenoids, and samples from Fujian contains more proteins. Between the 2 major varieties cultured in Zhejiang, Longzhi No. 1 contains higher content of triterpenoids, but Hunong No. 1 has more polysaccharides. Contents of triterpenoids and polysaccharides from wall-broken spores are much higher than those of fruiting bodies. The stipes from fruiting bodies contains more polysaccharides than those of the pileus, while the triterpenoids contents are higher in the pileus than stipes. The pileus and stipes collected in the second year contain higher content of polysaccharides than the first year's samples, but the contents of triterpenoids are lower. Wall-breaking treatment would significantly improve the extraction and dissolution rate of total triterpenoids and polysaccharides.

Fungal Proteins ; analysis ; Polysaccharides ; analysis ; Reishi ; chemistry ; Spores, Fungal ; chemistry ; Triterpenes ; analysis

Objective:To evaluate the clinical value of percutaneous transhepatic cholangial drainage (PTCD) and percutaneous transhepatic cholangial stent(PTCS)implantation therapy for malignant biliary obstruction in the elderly patients.Methods:Fifty three patients over 60 years-old were treated with PTCS implantation (n=35),PTCD for internal-external drainage(n=11),or single external drainage(n=7). The imaging examination was performed for all the patients one week after surgery. The serum levels of total bilirubin (TBIL) and alanine aminotransferase (ALT) were detected before and after surgery.Rusults:Metallic stents were successfully implanted in 32 patients during the surgery, and 3 patients were implanted 5 to 7 days later after PTCS. Both total serum levels of TBIL and ALT were significantly decreased at one week after surgery (P<0.05). The mean survival periods were 11.5 months for stent implantation patients and 5.5 months for PTCD patients. Conclusion:PTCD and PTCS are easy to perform and safe and effective in the treatment of malignant biliary obstruction of elderly patients.

Aging ; physiology ; Animals ; Female ; Male ; Mitochondria, Heart ; enzymology ; Mitochondrial Membrane Transport Proteins ; physiology ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism ; Ventricular Function, Left ; physiology

AIMTo study the effects and mechanism of naloxone on the febrile response in IL-1beta-induced fever rats.

METHODSThe fever model was established by intracerebroventricular injection of IL-1beta in rats. The effect of naloxone on the body temperature of feverrats was observed. The contents of cAMP in hypothalamus and AVP in VSA were detected.

RESULTSNaloxone alleviated IL-1beta-induced fever and the contents of cAMP in hypothalamus and AVP in VSA were correspondingly decreased (P < 0.01).

CONCLUSIONNaloxone could inhibit IL-1beta-induced fever in rats, and the mechanism might be due to inhibiting synthesis of cAMP in hypothalamus and promoting release of AVP in VSA.

Animals ; Arginine Vasopressin ; metabolism ; Cyclic AMP ; metabolism ; Fever ; metabolism ; Hypothalamus ; drug effects ; metabolism ; Male ; Naloxone ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Septum of Brain ; drug effects ; metabolism

OBJECTIVETo screen the point mutation of the low-density lipoprotein receptor (LDL-R) gene in Chinese familial hypercholesterolemia (FH) patients, characterize the relationship between the genotype and the phenotype and discuss the molecular pathological mechanism of FH.

METHODSA patient with clinical phenotype of homozygous FH and her parents were investigated for mutations in the promoter and all eighteen exons of the LDL-R gene. Screening was carried out using Touch-down PCR and direct DNA sequencing; multiple alignment analysis by DNASIS 2.5 was used to find base alteration, and the LDL-R gene mutation database was searched to identify the alteration. In addition, the apolipoprotein B gene (apo B) was screened for known mutations (R3500Q) that cause familial defective apo B100 (FDB) by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).

RESULTSTwo new heterozygous mutations in exons 4 and 9 of the LDL-R gene were identified in the proband (C122Y and T383I) as well as her parents. Both of the mutations have not been published in the LDL-R gene mutation database. No mutation of apo B100 (R3500Q) was observed.

CONCLUSIONTwo new mutations (C112Y and T383I) were found in the LDL-R gene, which may result in FH and may be particularly pathogenetic genotypes in Chinese people.

Adult ; Apolipoproteins B ; genetics ; Asian Continental Ancestry Group ; Child ; China ; Female ; Heterozygote ; Homozygote ; Humans ; Hyperlipoproteinemia Type II ; genetics ; Male ; Mutation ; Receptors, LDL ; genetics

OBJECTIVETo evaluate the ThinPrep bronchial brushing cytology in the diagnosis of lung cancers.

METHODSThe smear slides of ThinPrep bronchial brushing cytology in 1000 patients collected from April 2001 to April 2002 were reviewed.

RESULTSOf 389 patients diagnosed as having lung cancer clinically or histopathologically, 273 (70.2%) were revealed by ThinPrep bronchial brushing cytology. Among the 74 patients with benign lung diseases comfirmed by pathology, 3 had been suspected as having lung cancer by ThinPrep bronchial brushing cytology. Actually, they were two patients suffering from tuberculosis and one hamartoma proven by histopathology later. The sensitivity and specificity of ThinPrep bronchial brushing cytology were 70.2% and 95.9%, respectively. Of the 179 patients who had both cytological and histopathological results, the cytology and pathology concordance rate was 95.4% in squamous carcinoma, 87.0% in adenocarcinoma and 95.7% in small cell carcinoma.

CONCLUSIONAlthough ThinPrep bronchial brushing cytology has a high specificity, it is not good in diagnosing lung cancer. Poor smearing technique may be responsible for most of the false negative. Type diagnosis of poorly differentiated carcinomas can be difficult when based on the ThinPrep bronchial brushing cytology alone.

Adenocarcinoma ; diagnosis ; Adult ; Aged ; Biopsy ; Bronchi ; pathology ; Bronchoscopy ; Carcinoma, Small Cell ; diagnosis ; Carcinoma, Squamous Cell ; diagnosis ; Cytodiagnosis ; methods ; Diagnostic Errors ; Female ; Humans ; Lung Neoplasms ; diagnosis ; Male ; Middle Aged ; Sensitivity and Specificity

OBJECTIVETo study the inhibitory effect of paeoniflorin (PAE) on TNF-α-induced TNF receptor type I (TNFR1)-mediated signaling pathway in mouse renal arterial endothelial cells (AECs) and to explore its underlying molecular mechanisms.

METHODSMouse AECs were cultured in vitro and then they were treated by different concentrations PAE or TNF-α for various time periods. Expression levels of intercellular cell adhesion molecule-1 (ICAM-1) were detected in the normal group (cultured by serum-free culture media), the TNF-α group (cultured by 2-h serum-free culture media plus 6-h TNF-α 30 ng/mL), the low dose PAE group (cultured by 2-h PAE 0.8 μmo/L plus 6-h TNF-α 30 ng/mL), the middle dose PAE group (cultured by 2-h PAE 8 μmol/L plus 6-h TNF-α 30 ng/mL), the high dose PAE group (cultured by 2-h PAE 80 μmol/L plus 6-h TNF-α 30 ng/mL) with Western blot analysis. Nuclear translocation of transcription factor NF-κB (NE-κB) was detected in the normal group (cultured by serum-free culture media), the TNF-α group (cultured by 2-h serum-free culture media plus 45-mm TNF-α 30 ng/mL), and the high dose PAE group (cultured by 2-h PAE 80 μmol/L plus 45-min TNF-α 30 ng/mL) by immunofluorescent staining. Expression levels of the phosphorylation of extracellular signal-regulated (protein) kinase (ph-ERK) and p38 (ph- p38) were detected in the normal group (cultured by serum-free culture media) and the high dose PAE group (2-h PAE 80 μmol/L culture) by Western blot. NF-κB inhibitor-α (IκBα) protein expressions were detected in the normal group (cultured by serum-free culture media), the TNF-α group (cultured by 2-h serum-free culture media plus 30-min TNF-α 30 ng/mL), the high dose PAE group (cultured by 2-h PAE 80 μmol/L plus 30-min TNF-α 30 ng/mL), the p38 inhibitor group (SB group, pretreatment with SB238025 25 μmol/L for 30 min, then treated by PAE 80 μmol/L for 2 h, and finally treated by TNF-α 30 ng/mL for 30 min), the ERK inhibitor group (PD group, treated by PD98059 50 μmol/L for 30 min, then treated by PAE 80 μmol/L for 2 h, and finally treated by TNF-α 30 ng/mL for 30 min) by Western blot.

RESULTSCompared with the normal group, ICAM-1 protein expression levels obviously increased (P < 0.01). Compared with the TNFα group, ICAM-1 protein expression levels were obviously inhibited in the high dose PAE group (P < 0.05). Protein expression levels of ph-p38 and ph-ERK were obviously higher in the hIgh dose PAE group (P < 0.05). Compared with the normal group, IκBα protein expression levels obviously decreased in the TNF-α group (P < 0.01). Compared with the TNFα group, TNF-α-induced IκBα degradation could be significantly inhibited in the high dose PAE group (P < 0.01); the inhibition of PAE on IκBα degradation could be significantly inhibited in the SB group (P < 0.05). NF-κB/p65 signal was mainly located in cytoplasm in the normal group. NF-κB/p65 was translocated from cytoplasm to nucleus after stimulated by 45 min TNF-α in the TNF-α group, while it could be significantly inhibited in the high dose PAE group.

CONCLUSIONSPAE inhibited TNF-α-induced expression of lCAM-1. Its action might be associated with inhibiting TNFR1/NF-κB signaling pathway. p38 participated and mediated these actions.

Animals ; Cells, Cultured ; Endothelial Cells ; cytology ; drug effects ; Glucosides ; pharmacology ; Intercellular Adhesion Molecule-1 ; metabolism ; Mice ; Monoterpenes ; pharmacology ; NF-kappa B ; metabolism ; Receptors, Tumor Necrosis Factor ; metabolism ; Signal Transduction ; drug effects ; Tumor Necrosis Factor-alpha ; pharmacology

Porous carbon nanoparticles ( NPC) were prepared by ZnCl2 activation and carbonization using citrus waste as carbon source. A sample pretreatment method with NPC as dispersive solid phase extraction (d-SPE ) absorbent was established for the determination of organophosphorus pesticides in fruits and vegetables by gas chromatography. The NPC was characterized by scanning electron microscopy (SEM), X-ray diffraction ( XRD), FT-IR spectra, Raman spectroscopy, Brunauer, Emmett and Teller surface area(BET). Those results showed that the NPC was an amorphous porous carbon material with pore size in the range of 0-15 nm. Its specific surface area and pore volume were 1243 m2 / g and 1. 28 cm3 / g, respectively. The analysis conditions, including the amount and clean up time of adsorbent, were optimized by analysis of 14 kinds of oranophosphorus pesticides in fruits and vegetables with gas chromatography-flame photometric determination(GC-FPD). Moreover, the comparison for NPC with commercial materials of PSA, C18 and GCB was investigated in this study. The results indicated that the purification time was only 2 min using 0. 01 g NPC. The cost of NPC was about 25% of C18 , 21% of PSA and 16% of GCB. Because of the porous structure of NPC, the purification efficiency was significantly higher than the three commercial materials mentioned above. Under the optimum conditions, the calibration curves of the 14 organophosphorus pesticides were linear in the range of 0. 02-1. 00 mg / L with good correlation coefficients (R2>0. 99) and detection limits (S / N=3) of 0. 63-5. 30 μg / kg. The recoveries of the pesticides at three spiked levels ranged from 71. 3% to 114. 7%with the relative standard deviations (RSDs) of 0. 9% -12. 9% . The method is simple, rapid, sensitive, and low cost, and can satisfy the requirements of detection of organophosphorus pesticide residues in fruits and vegetables, displaying a good application prospect.