Objective To evaluate the feasibility of composite material of bone morphogenetic protein (BMP) and PLGA/TCP in repair of peri-porous-titanium bone defects in adult rabbits.Meth- otis The composite of PLGA/TCP scaffold with bovine BMP was made and implanted in distal bone de- fects peri-porous-titanium in femur of adult rabbits.The effect of BMP with PLGA/TCP on peri-prosthesis was evaluated by means of scanning electron microscope (SEM),energy dispersive X-ray analysis (EDX) and biomechanics.Results BMP with PLGA/TCP showed obvious peak of Ca and P of EDX in the pores of titanium in experiment group six weeks after operation and higher than that in control group (P＜0.05).Push-out test demonstrated that the bonding strength between the composite of HA coating/ porous titanium and bone was increased significantly with time (P＜0.05).In experiment group,at 6 and 12 weeks,peri-porous-titanium had higher shearing force compared with control group (P＜0.05). Conclusion BMP loaded with PLGA/TCP is a promising bone graft for bone defects in revision arthro- plasty,as indicates that bone induction of BMP plays an important role in biological stabilizaiton.

OBJECTIVEWe investigated the effects of pitavastatin on angiogenesis and perfusion in C3H/He mice with unilateral hind limb ischemia.

METHODSC3H/He mice treated with saline (n = 15) or pitavastatin (1 mg.kg(-1).d(-1), n = 15) per gavage for 1 week underwent unilateral hind limb ischemia surgery and were treated for another 5 weeks. Hind-limb blood flow was measured by Laser Doppler perfusion imager (LDPI, ischemic/nonischemic limb, %) at baseline, immediately after ischemia and weekly thereafter for 5 weeks. Endpoints included local vessel counts by immunofluorescence, phospho-Akt positive cell counts by immunoenzyme histochemical technique, vascular endothelial growth factors (VEGFs) expression in ischemic limbs by Western blot and serum nitric oxide metabolite (NOx) by chrome dioxide Griess method.

RESULTSLower extremity perfusion was significantly improved in pitavastatin treated mice vs. controls as measured by LDPI% at 1 week post ischemia and thereafter (P < 0.05). Pitavastatin treatment was associated with significantly increased capillary count [(47 +/- 11) vs. (26 +/- 14)/per high-power field (x 200), P < 0.05] and greater percentage of phospho-Akt positive cells [(6 +/- 1) vs. (2 +/- 0)/per high-power field (x 200), P < 0.05] in ischemic limbs. Serum NOx [(77.3 +/- 21.8) vs. (52.1 +/- 11.2) mol/L, P < 0.05) and VEGF protein expression in ischemic limbs were also significantly increased in pitavastatin group than those in control group.

CONCLUSIONSPitavastatin enhances angiogenesis and perfusion in CsH/He mice with limb ischemia.

Animals ; Disease Models, Animal ; Ischemia ; physiopathology ; Lower Extremity ; blood supply ; Male ; Mice ; Mice, Inbred C3H ; Neovascularization, Physiologic ; drug effects ; Nitric Oxide ; blood ; Quinolines ; pharmacology ; Vascular Endothelial Growth Factors ; metabolism

OBJECTIVETo construct the delta-pIRES2-EGFP plasmid and investigate its expression in HEK293 cells.

METHODSFull length cDNA of rat delta opioid receptor gene amplified from rat brain tissues using reverse transcription and nested PCR was cloned into pMD20 T vector. The delta cDNA was inserted into pIRES2-EGFP plasmid to construct the recombinant eukaryotic plasmid delta-pIRES2-EGFP, which was transfected into HEK293 cells via Lipofectamine2000. The expression of delta was examined under fluorescence microscope.

RESULTSThe recombinant delta-pIRES2-EGFP plasmid was successfully constructed, and high expression of delta was detected in HEK293 cells transfected by the plasmid.

CONCLUSIONdelta-pIRES2-EGFP has been successfully cloned, which shows high expression of delta in HEK293 cells.

Animals ; DNA, Complementary ; genetics ; Gene Expression ; Genetic Vectors ; Green Fluorescent Proteins ; biosynthesis ; genetics ; HEK293 Cells ; Humans ; Plasmids ; Polymerase Chain Reaction ; Rats ; Rats, Sprague-Dawley ; Receptors, Opioid, delta ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transfection

The total triterpene saponins of Psammosilene tunicoides have significant pharmacologic activity. Psammosilene tunicoides squalene synthase (PSS) is a gateway enzyme to regulate the biosynthesis of total triterpene saponins extracted from the root of Psammosilene tunicoides which is an endangered species. In this paper, cDNA encoding of PSS was cloned by the degenerate primer PCR and rapid-amplification of cDNA ends (RACE). The full-length of cDNA of PSS is 1663 bp, with an open reading frame (ORF) of 1 245 bp, encoding 414 amino acid polypeptide (calculated molecular mass, 47.69 kDa), 5'UTR (untranslated region) and 3'UTR are 260 bp and 158 bp, respectively. The deduced amino acid sequence of PSS has higher homology with the known squalene synthases of several species such as Panax notoginseng (83%), Panax ginseng (82%) and Glycyrrhiza glabra (82%) than that with Schizosacharomyces pombe (35%), Candida albicans (39%) and Homo sapiens (47%). The characterization of PSS was done by a series of methods, such as prokaryotic expression, the activity of enzyme in vitro, capillary gas chromatography (GC) and capillary gas chromatography mass spectrometry (GC-MS). The results showed that the cell-free extract of E. coli transformed with the recombinant plasmid can effectively convert farnesyl diphosphate into squalene in vitro. GenBank accession number is EF585250. Our research provided important base for the study of Psammosilene tunicoides secondary metabolism and metabolic engineering.
Caryophyllaceae ; enzymology ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; Endangered Species ; Escherichia coli ; genetics ; metabolism ; Farnesyl-Diphosphate Farnesyltransferase ; genetics ; metabolism ; Gas Chromatography-Mass Spectrometry ; Open Reading Frames ; Phylogeny ; Plant Proteins ; genetics ; metabolism ; Plants, Medicinal ; chemistry ; genetics ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; metabolism ; Sequence Homology, Amino Acid ; Transformation, Genetic

Objective To compare the effects of 20 mg qd and 10 mg bidadministration of iprrazole enteric-coated tablets on the control of gastric acid in healthy subjects.Methods A randomized,single-center,parallel controlled trial was designed to include 8 healthy subjects.Randomly divided into 2 groups,20 mg qd administration group:20 mg enteric-coated tablets of iprrazole in the morning;10 mg bid administration group:10 mg enteric-coated tablets of iprrazole in the morning and 10 mg in the evening.The pH values in the stomach of the subjects before and 24 h after administration were monitored by pH meter.The plasma concentration of iprazole after administration was determined by HPLC-MS/MS.The main pharmacokinetic parameters were calculated by Phoenix WinNonlin(V8.0)software.Results The PK parameters of iprrazole enteric-coated tablets and reference preparations in fasting group were as follows:The Cmax of 20 mg qd group and 10 mg bid group were(595.75±131.15)and(283.50±96.98)ng·mL-1;AUC0-t were(5 531.94±784.35)and(4 686.67±898.23)h·ng·mL-1;AUC0-∞ were(6 003.19±538.59)and(7 361.48±1 816.77)h·ng·mL-1,respectively.The mean time percentage of gastric pH＞3 after 20 mg qd and 10 mg bid were 82.64％and 61.92％,and the median gastric pH within 24 h were 6.25±1.49 and 3.53±2.05,respectively.The mean gastric pH values within 24 h were 5.71±1.36 and 4.23±1.45,respectively.The correlation analysis of pharmacokinetic/pharmacodynamics showed that there was no significant correlation between the peak concentration of drug in plasma and the inhibitory effect of acid.Conclusion Compared with the 20 mg qd group and the 10 mg bid group,the acid inhibition effect is better,the administration times are less,and the safety of the two administration regimes is good.

OBJECTIVETo apply echocardiography in diagnosis of isolated noncompaction of ventricular myocardium (INVM) in adults.

METHODSSix patients with INVM underwent echocardiographic examination using HP5500 ultrasound system with the frequency of 2 approximately equals 4 MHz, and the observation was focused on ventricularmyocardium and endocardium at one-third of the apex.

RESULTSAll 6 patients showed typical echocardiographic images characterized by an altered structure of the ventricular myocardium with extremely thickened, hypokinetic segments consisting of two layers: a thin, compacted epicardium and an extremely thickened endocardium, resulting in an extremely thickened ventricular wall.

CONCLUSIONEchocardiography should be the first choice to be applied in diagnosis of INVM, which is a distinct entity of specific cardiomyopathy in adults.

Adult ; Cardiomyopathies ; congenital ; diagnostic imaging ; pathology ; Echocardiography, Doppler, Color ; Female ; Heart Ventricles ; diagnostic imaging ; pathology ; Humans ; Male ; Middle Aged ; Myocardium ; pathology

OBJECTIVETo assess the metastatic frequency in different groups of lymph nodes and its influencing factors of the thoracic esophageal squamous cell carcinoma (ESCC) in order to determine the extent of lymphadenectomy during esophagectomy.

METHODSThe clinical data of 730 patients with ESCC who underwent esophagectomy and lymphadenectomy were analyzed retrospectively.

RESULTSOf 730 patients, 166 had metastasis to the para-esophageal lymph nodes (22.7%), 90 to the left gastric artery lymph nodes (12.3%), 67 to the lymph nodes around gastric cardia, and 15 to the subcrinal lymph nodes (2.1%). Univariate analysis showed that metastasis to the subcrinal lymph node was positively correlated with the length and differentiation of tumor (P < 0.05), but it was not correlated with any the above parameters when analyzed by multivariate analysis. The metastasis to the para-esophageal lymph node was positively correlated with the length, invasion depth and differentiation of tumor by univariate and multivariate analysis (P < 0.05). The metastasis to the lymph nodes around gastric cardia and metastasis to left gastric artery lymph nodes were positively correlated with the position and invasion depth of tumor by univariate and multivariate analysis (P < 0.05).

CONCLUSIONLymph nodes of the para-esophagus, gastric cardia and left gastric artery usually have high frequency to harber mestastasis, therefore, it was suggested that the lymph nodes in these groups should be dissected during esophagectormy with two-field lymphadenectomy for thoracic esophageal squamous cell carcinoma. Whereas for those patients with the lesion < 3 cm in length or with tumor invasion confined within the esophageal wall or with a lesion located at the upper or lower third of the thoracic esophagus, the subcrinal lymph nodes may not be necessarily dissected.

Adult ; Aged ; Carcinoma, Squamous Cell ; pathology ; surgery ; Cardia ; Esophageal Neoplasms ; pathology ; surgery ; Esophagectomy ; methods ; Esophagus ; Female ; Humans ; Lymph Node Excision ; methods ; Lymph Nodes ; pathology ; surgery ; Lymphatic Metastasis ; Male ; Middle Aged ; Multivariate Analysis ; Neoplasm Invasiveness ; Neoplasm Staging ; Retrospective Studies

OBJECTIVETo observe and analyse the dissection of the tunnels in traditional blind operation of augmentation rhinoplasty.

METHODS11 Cases of augmentation rhinoplasty were collected and be observed by an endoscope as soon as the tunnels were formed during the operations.

RESULTS(1) Some of the tunnels did not go through one layer. (2) The bilateral cartilage separated in the mid-line. (3) There were two blood vessels in the surface of alar cartilage. There were perforating blood vessels in the edge of pyriform aperture. (4) In some cases whose incision were in unilateral alar margin, the tunnel were asymmetric.

CONCLUSIONIn some cases of traditional blind operation of augmentation rhinoplasty, tunnels were not suitable, they were asymmetric; and there were desmo and septa in the tunnels. Those might be the causes of complications post-op of augmentation rhinoplasty.

Endoscopes ; Humans ; Nose ; anatomy & histology ; Rhinoplasty ; methods

OBJECTIVETo investigate the correlation of the mRNA expression level of excision repair cross-complementing group 1 (ERCC1) gene with clinicopathological parameters and clinical outcome in patients with non-small cell lung cancer (NSCLC) receiving platinum-based chemotherapy.

METHODSThe mRNA expression of ERCC1 in formalin-fixed paraffin-embedded primary tumor specimens was measured by real-time quantitative reverse transcriptase polymerase chain reaction. The association between ERCC1 expression levels and clinicopathological parameters in NSCLC patients was analyzed.

RESULTSThe median value of ERCC1 mRNA expression level compared with beta-actin in tumor specimens of 61 NSCLC patients was 0.48. There was no correlation between ERCC1 expression and clinicopathological parameters. Patients with low expression of ERCC1 mRNA (less than 0.35, 0.28, respectively) had a significantly longer median time to progression (TTP) (14.3 vs. 8.0 months, P = 0.028) and overall survival (OS) (28.4 vs. 12.9 months, P = 0.0064) than those with high expression. Multivariate analysis showed that a low ERCC1 mRNA expression was an independent factor for OS.

CONCLUSIONOur findings suggest that intratumoral ERCC1 mRNA expression level, although is uncorrelated with clinicopathological parameters, is an independent predictive marker for survival of the patients with NSCLC receiving platinum-based chemotherapy, and may provide critical information for personalized chemotherapy.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Bone Neoplasms ; secondary ; Brain Neoplasms ; secondary ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; metabolism ; pathology ; secondary ; Cisplatin ; administration & dosage ; DNA-Binding Proteins ; genetics ; metabolism ; Disease-Free Survival ; Endonucleases ; genetics ; metabolism ; Female ; Follow-Up Studies ; Humans ; Lung Neoplasms ; drug therapy ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Paraffin Embedding ; Platinum ; administration & dosage ; Proportional Hazards Models ; RNA, Messenger ; metabolism ; Survival Rate

OBJECTIVETo explore the eukaryotic expression of arresten in CHO cells and to investigate its basic biological activities.

METHODSCHO cells were divided into three groups: transfected pSecTag-arresten group, transfected pSecTag group and control group without transfection. PSecTag-arresten was transfected into CHO cells by Lipofectamine 2000 method. The arresten mRNA in CHO cells was assayed by RT-PCR. The protein expression of arresten gene was examined by Western-Blot. The cells expressing arresten were screened out by Zeocin. The effect of arresten on huvec cell migration and anchoring to three-dimensional vascular structures was measured.

RESULTSThe result of RT-PCR and Western-blot showed that arresten gene has been successfully transfected into CHO cells and expressed in those cells. Arrssten inhibited huvec cell migration and anchoring to three-dimensional vascular structures.

CONCLUSIONCHO cells expressing arresten have been obtained successfully. Arresten can inhibit huvec cell migration and anchoring to three-dimensional vascular structures, indicating that it might be one of its anti-angiogenetic approaches.

Angiogenesis Inhibitors ; biosynthesis ; genetics ; pharmacology ; Animals ; Blotting, Western ; CHO Cells ; Cell Line ; Cell Movement ; drug effects ; Cells, Cultured ; Collagen Type IV ; biosynthesis ; genetics ; pharmacology ; Cricetinae ; Cricetulus ; Endothelial Cells ; cytology ; drug effects ; physiology ; Humans ; Neovascularization, Physiologic ; drug effects ; RNA, Messenger ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection