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The magnitude of inbreeding depression within populations is important for the evolution and maintenance of mixed mating systems. However, data are sparse on the magnitude of inbreeding depression in Robinia pseudoacacia. In this study, we compared differences in the mature seed set per fruit, seed mass, germination success, and seedling growth between self- and cross-pollination treatments and estimated the inbreeding depression at 3 stages: seed maturation, seedling emergence, and seedling growth at 10 and 20 weeks. We found that progenies resulting from cross-pollination treatments showed significantly higher fitness than progenies resulting from self-pollination, causing high levels of inbreeding depression. Inbreeding depression was not uniformly manifested, however, over the 3 stages. Inbreeding depression was the greatest between fertilization and seed maturation stage (δ = 0.5419), and the seedling emergence (0.3654) stage was second. No significant differences in seedling growth were observed between selfed and crossed progenies. The cumulative inbreeding depression (δ) across all 3 stages averaged 0.7452. Inbreeding depression may promote outcrossing in R. pseudoacacia by acting as a post-pollination barrier to selfing. The large difference in the seed set between self- and cross-pollination that we detected indicated that inbreeding depression would probably be a reasonable explanation for the high abortion and low seed set in R. pseudoacacia.

The synthesis of benzimidazoles by intermolecular cyclization reaction of 2-iodoanilines with nitriles has been developed. These reactions proceeded without the aid of any transition metals or ligands and just using KOBu(t) as the base. A variety of substituted benzimidazole derivatives can be synthesized by the approach.

The grasshoppers are ideal materials to study various meiotic stages of spermatogenesis due to their easy availability, fairly large chromosomes, and fewer numbers of chromosomes. It is easy to make temporary squash preparation of grasshopper testes; however, it is usually difficult for the beginners to differentiate between stages of meiosis. In view of this, we demonstrated the method of identification of meiotic stages by chromosome number and chromosome conformation, taking spermatogonial meiosis of Locusta migratoria manilensis as an example. We described briefly the mitosis of spermatogonia and the spermatogenesis of this species as well.

Transforming growth factor (TGF)-β and activin, members of TGF-β superfamily, are abundantly expressed in the endometrium and regulate decidualization of endometrial stroma. Smad2 and Smad3 are receptor-regulated Smads (R-Smads) that transduce extracellular TGF-β/activin/Nodal signaling. In situ hybridization results showed that Smad3 was highly expressed in the decidual zone during the peri-implantation period in mice. By using artificial decidualization, we found that Smad3 null mice showed partially compromised decidualization. We therefore hypothesized that Smad2 might compensate for the function of Smad3 during the process of decidualization. Smad2 was also highly expressed in the decidual zone and phosphorylated Smad2 was much more abundantly increased in the deciduoma of Smad3 null mice than for wild-type (WT) mice. We further employed an in vitro uterine stromal cell decidualization model, and found that decidual prolactin-related protein (dPRP) and cyclin D3, which are well-known markers for decidual cells, were significantly down-regulated in Smad3 null decidual cells, and were much more significantly reduced when the expression of Smad2 was simultaneously silenced by its siRNA (P < 0.05). However, the expression levels of dPRP and cyclin D3 remained the same when Smad2 was silenced in WT decidual cells. Collectively, these findings provide evidence for an important role of Smad3 in decidualization and suggest that Smad2 and Smad3 may have redundant roles in decidualization., (Copyright © 2012 Wiley Periodicals, Inc.)

Our purpose was to evaluate the effect of the mechanical force of a sneeze on sinonasal cilia function and determine the molecular mechanism responsible for eliciting the ciliary response to a sneeze. A novel model was developed to deliver a stimulation simulating a sneeze (55 mmHg for 50 ms) at 26°C to the apical surface of mouse and human nasal epithelial cells. Ciliary beating was visualized, and changes in ciliary beat frequency (CBF) were determined. To interrogate the molecular cascades driving sneeze-induced changes of CBF, pharmacologic manipulation of intra- and extracellular calcium, purinergic, PKA, and nitric oxide (NO) signaling were performed. CBF rapidly increases by ≥150% in response to a sneeze, which is dependent on the release of adenosine triphosphate (ATP), calcium influx, and PKA activation. Furthermore, apical release of ATP is independent of calcium influx, but calcium influx and subsequent increase in CBF are dependent on the ATP release. Lastly, we observed a blunted ciliary response in surgical specimens derived from patients with chronic rhinosinusitis compared to control patients. Apical ATP release with subsequent calcium mobilization and PKA activation are involved in sinonasal ciliary response to sneezing, which is blunted in patients with upper-airway disease.

Semi-one-pot synthesis of phenanthrenes from styrenes and arenes was developed through cross-dehydrogenative coupling. A sequence of Heck-type coupling and photo-cyclization were involved and a variety of functionalities were tolerated. This method provides an effective and practical protocol towards the synthesis of substituted phenanthrenes.

Two different tetrasubstituted benzenes were selectively constructed from the same starting materials by tuning the active sites of the Grubbs second generation catalyst (GC-II) with CuI or AgOTf as the additive., (This journal is © The Royal Society of Chemistry 2012)

Mucociliary clearance is the primary innate physical defense mechanism against inhaled pathogens and toxins. Vectorial ion transport, primarily sodium absorption and anion secretion, by airway epithelial cells supports mucociliary clearance. This is evidenced by diseases of abnormal ion transport such as cystic fibrosis and pseudohypoaldosteronism that are characterized by changes in mucociliary clearance. Sodium absorption and chloride secretion in human bronchial epithelial cells depend on potassium channel activity, which creates a favorable electrochemical gradient for both by hyperpolarizing the apical plasma membrane. Although the role of basolateral membrane potassium channels is firmly established and extensively studied, a role for apical membrane potassium channels has also been described. Here, we demonstrate that bupivacaine and quinidine, blockers of four-transmembrane domain, two-pore potassium (K2P) channels, inhibit both amiloride-sensitive sodium absorption and forskolin-stimulated anion secretion in polarized, normal human bronchial epithelial cells at lower concentrations when applied to the mucosal surface than when applied to the serosal surface. Transcripts from four genes, KCNK1 (TWIK-1), KCNK2 (TREK-1), KCNK5 (TASK-2), and KCNK6 (TWIK-2), encoding K2P channels were identified by RT-PCR. Protein expression at the apical membrane was confirmed by immunofluorescence. Our data provide further evidence that potassium channels, in particular K2P channels, are expressed and functional in the apical membrane of airway epithelial cells where they may be targets for therapeutic manipulation.

Background: Cilia are complex and powerful cellular structures of the respiratory mucosa that play a critical role in airway defense. Respiratory epithelium is lined with cilia that perform an integrated and coordinated mechanism called mucociliary clearance. Mucociliary clearance is the process by which cilia transport the mucus blanket overlying respiratory mucosa to the gastrointestinal tract for ingestion. It is the primary means by which the airway clears pathogens, allergens, debris, and toxins. The complex structure and regulatory mechanisms that dictate the form and function of normal cilia are not entirely understood, but it is clear that ciliary dysfunction results in impaired respiratory defense., Methods: A literature review of the current knowledge of cilia dysfunction in chronic rhinosinsusitis was conducted., Results: Ciliary dysfunction may be primary, the result of genetic mutations resulting in abnormal cilia structure, or, more commonly, secondary, the result of environmental, infectious, or inflammatory stimuli that disrupt normal motility or coordination. Patients with chronic rhinosinusitis (CRS) have been found to have impaired mucociliary clearance. Many biochemical, environmental, and mechanical stimuli have been shown to influence ciliary beat frequency, and common microbial pathogens of respiratory mucosa such as Pseudomonas aeruginosa and Haemophilus influenzae have developed toxins that appear to interrupt normal mucociliary function. Furthermore, inflammatory mediators known to be present in patients with CRS appear to impair secondarily mucociliary clearance., Conclusion: The goal of this article is to summarize the recent developments in the understanding of cilia dysfunction and mucociliary clearance in CRS.

Background: Sinonasal mucosal biofilms are recognized as contributors to the pathogenesis of chronic rhinosinusitis (CRS). Attachment of bacteria to the sinonasal surface is an initial step in biofilm formation. A critical defense against this occurrence is mucociliary clearance (MCC). To ascertain whether the ciliary component of MCC is uniform throughout the airway we compared ciliary beat frequency (CBF) in the murine nasal septum and trachea at baseline and after challenge with Pseudomonas aeruginosa, a common pathogen of CRS., Methods: Murine septal and tracheal air-liquid interface cultures were evaluated for basal and stimulated CBF after exposure to control or conditioned media from Pseudomonas. Additionally, the attachment of Pseudomonas to nasal and tracheal cultures was assessed after pretreatment with control or conditioned media., Results: Basal CBF is significantly slower in primary nasal airway cultures compared with tracheal airway cultures. Tracheal airway cultures show resistance to Pseudomonas secreted ciliotoxins not evident in nasal septal cultures. Furthermore, after challenge with viable Pseudomonas, significantly more bacteria attach to the nasal cultures compared with the tracheal cultures., Conclusion: Using primary murine nasal and tracheal airway cultures we show inherent differences in cilia function and increased susceptibility of the upper airway to attachment by Pseudomonas. Understanding the differences between upper and subglottic airway mucociliary clearance should lead to novel approaches in the management of upper airway infection.

A new truxene discotic liquid crystal possessing only three octyloxy chains (3C8OTRX) was studied on the mesomorphic and semiconducting properties to reveal that it exhibits a high drift mobility of positive carriers in the order of 10(-2) cm(2) V(-1) s(-1) in the hexagonal ordered columnar (Col(ho)) mesophase., (This journal is © The Royal Society of Chemistry 2011)

Cigarette smokers and those exposed to second hand smoke are more susceptible to life threatening infection than non-smokers. While much is known about the devastating effect tobacco exposure has on the human body, less is known about the effect of tobacco smoke on the commensal and commonly found pathogenic bacteria of the human respiratory tract, or human respiratory tract microbiome. Chronic rhinosinusitis (CRS) is a common medical complaint, affecting 16% of the US population with an estimated aggregated cost of $6 billion annually. Epidemiologic studies demonstrate a correlation between tobacco smoke exposure and rhinosinusitis. Although a common cause of CRS has not been defined, bacterial presence within the nasal and paranasal sinuses is assumed to be contributory. Here we demonstrate that repetitive tobacco smoke exposure induces biofilm formation in a diverse set of bacteria isolated from the sinonasal cavities of patients with CRS. Additionally, bacteria isolated from patients with tobacco smoke exposure demonstrate robust in vitro biofilm formation when challenged with tobacco smoke compared to those isolated from smoke naïve patients. Lastly, bacteria from smoke exposed patients can revert to a non-biofilm phenotype when grown in the absence of tobacco smoke. These observations support the hypothesis that tobacco exposure induces sinonasal biofilm formation, thereby contributing to the conversion of a transient and medically treatable infection to a persistent and therapeutically recalcitrant condition.

Cartilage has a poor intrinsic repair capacity, requiring surgical intervention to effect biological repair. Tissue engineering technologies or regenerative medicine strategies are currently being employed to address cartilage repair. Mesenchymal stem cells (MSCs) are considered to be an excellent cell source for this application. However, the different gene expression profiles between the MSCs and differentiated cartilage remain unclear. In this report, we first examined the gene expression profiles between the MSCs, hyaline and elastic chondrocytes, and then identify candidate genes, which may be important in the process of MSC differentiation into hyaline and elastic cartilage. Several hundred differentially expressed genes were screened initially by microarray, including 417 simultaneously up-regulated genes in both hyaline and elastic chondrocytes, with 313 down-regulated genes. Several genes were identified that were up-regulated in hyaline chondrocytes while down-regulated in elastic chondrocytes. Both RT-PCR and western blot analysis were consistent with those results obtained by microarray analysis. Chondromodulinl (Chm1) was found to be highly expressed in MSCs differentiating to hyaline and elastic cartilage. Both collagen type II, alpha 1 (Col2a1) and cartilage homeo protein 1 (Cart1) were also highly upregulated and may be important early differentiation of MSCs to hyaline cartilage.

Wheat germ cell-free methods provide an important approach for the production of eukaryotic proteins. We have developed a protein expression vector for the TNT((R)) SP6 High-Yield Wheat Germ Cell-Free (TNT WGCF) expression system (Promega) that is also compatible with our T7-based Escherichia coli intracellular expression vector pET15&#95;NESG. This allows cloning of the same PCR product into either one of several pET&#95;NESG vectors and this modified WGCF vector (pWGHisAmp) by In-Fusion LIC cloning (Zhu et al. in Biotechniques 43:354-359, 2007). Integration of these two vector systems allowed us to explore the efficacy of the TNT WGCF system by comparing the expression and solubility characteristics of 59 human protein constructs in both WGCF and pET15&#95;NESG E. coli intracellular expression. While only 30% of these human proteins could be produced in soluble form using the pET15&#95;NESG based system, some 70% could be produced in soluble form using the TNT WGCF system. This high success rate underscores the importance of eukaryotic expression host systems like the TNT WGCF system for eukaryotic protein production in a structural genomics sample production pipeline. To further demonstrate the value of this WGCF system in producing protein suitable for structural studies, we scaled up, purified, and analyzed by 2D NMR two (15)N-, (13)C-enriched human proteins. The results of this study indicate that the TNT WGCF system is a successful salvage pathway for producing samples of difficult-to-express small human proteins for NMR studies, providing an important complementary pathway for eukaryotic sample production in the NESG NMR structure production pipeline.

Functional protein analysis often calls for lengthy, laborious in vivo protein expression and purification, and can be complicated by the lack of stability of the purified protein. In this study, we demonstrate the feasibility of a simplified procedure for functional protein analysis on magnetic particles using cell-free protein synthesis of the catalytic subunit of human cAMP-dependent protein kinase as a HaloTag((R)) fusion protein. The cell-free protein synthesis systems provide quick access to the protein of interest, while the HaloTag technology provides efficient, covalent protein immobilization of the fusion protein, eliminating the need for further protein purification and minimizing storage-related stability issues. The immobilized cPKA fusion protein is assayed directly on magnetic beads and can be used in inhibitor analyses. The combination of rapid protein synthesis and capture technologies can greatly facilitate the process of protein expression and activity screening, and therefore, can become a valuable tool for functional proteomics studies.

Objective: To investigate the etiology of Budd-Chiari syndrome (BCS) preliminarily., Methods: The clinical findings of radical surgery of 109 cases with BCS from March 2001 to May 2009 were analyzed. The pathological components of membranous tissue (MT) from inferior vena cava (IVC) or hepatic vein (HV) of BCS patients were compared with that of thrombus from deep venous thrombosis (DVT), as well as the expression of transforming growth factor beta receptor (TGF-beta R), platelet derived growth factor receptor (PDGFR), endothelin (ET-1), factor VIII related antigen (FVIII-rAg), ferritin and alpha1-antitrypsin in MTs and thrombus through immunohistochemical method., Results: One hundred and four cases of BCS were due to IVC and/or HV membrane or thrombosis except that 4 cases due to IVC tumor or 1 case due to compression of fiber. The new-formed IVC membrane was found in 2 recurred cases whose IVC thrombus was excised before 1 year and 7 years. The development from organized thrombus to MT was found in 3 cases of segmental obstruction of IVC. The IVC membrane located below HV outlet was in 8 cases. Both MTs and thrombus had the pathological components such as fibroblast, neutrophil, granulation tissue, newly-formed blood vessels and so on under the light microscope. The expressions of TGF-beta R, PDGFR, ET-1, FVIII-rAg, and ferritin in MTs and thrombus were as follows: MT 72.3%, thrombus 50.0% (P > 0.05); MT 45.5%, thrombus 100% (P < 0.05); MT 100%, thrombus 0 (P < 0.05); MT 90.9%, thrombus 12.5% (P < 0.05); MT 72.3%, thrombus 100% (P > 0.05)., Conclusions: The membranous tissues and thrombus have the similar homogeneity and cytokines expression. The membrane and thrombus may be different pathological phases.

Suzuki-Miyaura coupling of unactivated alkenyl carbamates is described to construct polysubstituted olefins. The developed process is also suitable for heteroaromatic and even electron-rich aromatic carbamates.

Objective: To analyze the relative factors of early-term restenosis after artificially grafting bypasses on chronic ischemia of lower extremities., Methods: From January 2006 to September 2007, 40 cases suffered from chronic ischemia of lower extremities were treated by single side femoropopliteal bypass were followed up during 6 months after operation. There were 36 male and 4 case female with a mean age of (66 +/- 9) years old. Lipid, fibrinogen (FIB) and hypersensitive C reactive protein (hsCRP) were chemical examined during peri-operation. Basing on the degree of restenosis in vascular anastomosis by Color Doppler graft scan, all the patients were divided into light, moderate and severe groups, respectively. Biochemical indicators and cytokines were investigated such as lipid, FIB, hsCRP, IL-6, transforming growing factor beta1 (TGF-beta1). Possible risk factors resulting in restenosis were compared statistically among three groups with SPSS 15.0., Results: Restenosis were more severe among the patients with concomitance disease such as diabetes mellitus and smoking after operation. Relative risk were 6.47 and 7.92, respectively. There are significant difference in total cholesterin, low density lipoprotein, FIB, hsCRP, IL-6 and TGF-beta1 among three groups during six months after operation (P < 0.05). Multiple linear regression showed that FIB and TGF-beta1 may be the risk factors to intimal hyperplasia., Conclusion: Diabetes mellitus, smoking and higher levels of FIB may be the major high risk factors resulting in neointima hyperplasia and anastomosis restenosis.

Objective: To find the related syndrome factors of gouty arthritis with blood stasis syndrome by mathematical methods of set pair analysis and connection number so as to guide the diagnosis., Methods: Seventy gouty arthritis patients diagnosed with blood stasis syndrome were analyzed according to priority by the method of connection number. Syndrome factors of gouty arthritis with blood stasis syndrome were selected according to the numerical values of dialectics connection value (DCV)., Results: In the nine observed items, the DCVs of five syndrome factors were above 0.5. These five factors were pain (0.858 0), sublingual varices and positive signs (varices and dark purple color) for palatal mucosa (0.773 1), tophus (0.723 0), varicose veins and telangiectasis (0.700 9), and squamous and dry skin (0.612 3). The DCVs of the other factors were below 0.5., Conclusion: The main factor 1 is pain; factor 2 is sublingual varices and positive signs (varices and dark purple color) for palatal mucosa; factor 3 is tophus; factor 4 is varicose veins and telangiectasis; factor 5 is dry skin. The others are secondary factors.

For protein microarrays, maintaining protein stability during the slide processing steps of washing, drying, and storage is of major concern. Although several studies have focused on the stability of immobilized antibodies in antibody microarrays, studies on protein-protein interaction arrays and enzyme arrays are lacking. In this paper we used five bait-prey protein interaction pairs and three enzymes to optimize the washing, drying, and storage conditions for protein arrays. The protein arrays for the study were fabricated by combining HaloTag technology and cell-free protein expression. The HaloTag technology, in combination with cell-free expression, allowed rapid expression and immobilization of fusion proteins on hydrogel-coated glass slides directly from cell extracts without any prior purification. Experimental results indicate enzyme captured on glass slides undergoes significant loss of activity when washed and spin-dried using only phosphate buffer, as is typically done with antibody arrays. The impact of washing and spin-drying in phosphate buffer on protein-protein interaction arrays was minimal. However, addition of 5% glycerol to the wash buffer helps retain enzyme activity during washing and drying. We observed significant loss of enzyme activity when slides were stored dry at 4 degrees C, however immobilized enzymes remained active for 30 days when stored at -20 degrees C in 50% glycerol. We also found that cell-free extract containing HaloTag-fused enzymes could undergo multiple freeze/thaw cycles without any adverse impact on enzyme activity. The findings indicate that for large ongoing studies, proteins of interest expressed in cell-free extract can be stored at -70 degrees C and repeatedly used to print small batches of protein array slides to be used over a few weeks.

Objective: To report the mid- to long-term outcome of artery bypass in chronic ischemia of lower extremities., Methods: The clinical data of 212 patients treated with bypass operation from January 2002 to April 2007 were retrospectively reviewed. Femoropopliteal artery bypass to above-knee popliteal (FP-ak) was carried out in 111 cases, femoropopliteal artery bypass to below-knee popliteal (FP-bk) in 59 cases, aortoiliac bypass in 25 cases and femorofemoral bypass in 17 cases., Results: One hundred and eighty-six patients (87.7%) were followed up for 6 to 68 months (median, 18 months). One-year primary patency rate of FP-ak and FP-bk was 69.7% and 53.5%, respectively. After graft revision, 1-year secondary patency rate of FP-ak and FP-bk were 81.6% and 60.5%, respectively. The 3-year patency with FP-ak (56.3%) was significantly higher than that in FP-bk (23.8%) (P < 0.05). Fifty-two cases were reoperated on during the follow-up period. Crural or femoral amputation could not avoid in 23 cases (limb salvage rate 89.2%). Ten cases died in 1 to 30 days after the operation, 20 cases died later during followup, and most of them died of cardio-cerebrovascular diseases. Artificial vessel infection occurred in 6 cases., Conclusions: The selection of surgical treatment for chronic ischemia of lower extremities should based on the ischemic state of the limb. The mid- to long-term patency rate of FP-ak is higher than that of FP-bk.

Theoretical studies on 1,4-bis [(p-methoxyphenoxy) carbonyl] and ramification were carried out using the density functional theory at B3LYP/6-31+G* level to obtain optimized equilibrium structure, vibrational spectra and electronic spectra. The calculation results indicate that the carbon and oxygen (in the ester group) come into being different large pi bonds with the benzene circles because of place blocking and conjugate effect. On this basis, the first excited state electronic transition energy was calculated by time-dependent density function theory. The calculated results show that the largest absorption spectra of the title compounds were obtained from the pi --> pi* electron transition from the highest occupied molecular orbital to the lowest unoccupied molecular orbital. And lambda(max) was obtained in 370-384 nm, belonging to the UV absorption area. The computed results show that the lateral substitutent of 1,4-bis [(p-methoxyphenoxy) carbonyl] has little effects on molecular structure. Because of place blocking, the dihedral angle between benzene (1) and benzene (2) augments 3 degrees-4 degrees. The introduction of the lateral substitutents has effects on vibrational spectra. The results showed that with the introduction of hydroxy the difference between E(HOMO) and E(LUMO) of the compound becomes small. While lambda(max) becomes large. Meanwhile, the difference between E(HOMO) and E(LUMO) reduces by 0.120 9 eV with the introduction of fluorin. This causes the biggest absorption wavelength to red shift 14 nm.

Aryl C-O and C-F can be transformed into C-Me via Ni-catalyzed coupling with MeMgBr under mild conditions.

The Leader protein is a defining feature of picornaviruses from the Cardiovirus genus. This protein was recently shown to inhibit cellular nucleocytoplasmic transport through an activity mapped to its zinc-binding region. Here we report the three-dimensional solution structure determined by nuclear magnetic resonance (NMR) spectroscopy of this domain (residues 5-28) from mengovirus. The domain forms a CHCC zinc-finger with a fold comprising a beta-hairpin followed by a short alpha-helix that can adopt two different conformations. This structure is divergent from those of other eukaryotic zinc-fingers and instead resembles motifs found in a group of DNA-binding proteins from Archaea.

Two complementary classes of molecules based on a triphenylene core are synthesized. The two-dimensional (2D) assemblies of these molecules deposited on a highly oriented pyrolytic graphite (HOPG) surface are identified with scanning tunneling microscopy (STM). Structures with large cavities are formed by symmetric molecules, while uniform and closely packed stripe-assembled structures are obtained for asymmetric molecules. X-ray diffraction (XRD) results support the observation of an ordered hexagonal columnar mesophase for symmetric molecules and a rectangular columnar mesophase for asymmetric molecules. The study demonstrates that the substitution symmetry has significant effects on the assembly characteristics of molecular architectures and also on the three-dimensional (3D) macroscopic properties of the molecular materials.

Wheat germ based eukaryotic cell-free systems have been shown to be applicable for both functional and structural analyses of proteins. However, the existing methods might require specialized instrumentation and/or a separate mRNA synthesis step. We have developed a DNA based, highly productive, coupled transcription/translation wheat germ cell-free system that incorporates the normally separate mRNA synthesis step and does not require specialized instrumentation. Using a small-volume batch reaction with fluorescence labeling, DNA templates predicted to encode proteins could be quickly screened for their ability to direct the expression of proteins of the appropriate size. Protein yield can be increased as much as 2 to 4-fold in this system using a dialysis reaction, reaching approximately 200-440 microg/ml in 10-20 h. Furthermore, enzyme activities can be assayed directly in the extract without further purification. Simple purification with affinity tags can be achieved in one-step and with minor modifications, efficient SeMet and [U-15N] labeling of >95% can be accomplished in this system. Thus, this efficient cell-free expression system can facilitate both functional and structural proteomics.

Objective: To develop a new way to prevent restenosis in the anastomotic site due to intimal hyperplasia after vascular graft bypass (VGB) in peripheral arteries., Methods: Five mongrel dogs received bilateral iliac-femoral VGB and their arteries between the graft were ligated and cut off under general anaesthesia. The mixture of the paclitaxel and fibrin gel (FG) were randomly sprayed onto one side of grafts including distal and proximal anastomotic site, and the fibrin gel served as control were sprayed onto the other one. The bilateral grafts including distal and proximal anastomotic site were harvested four weeks postoperationally and the anastomotic sites were observed grossly, pathologically and by electron microscopy. The intimal thickness and area of each anastomotic site were measured, then the data were analysed statistically., Results: The bilateral grafts of all dogs were patent and the neointima of all anastomotic sites have been seen grossly. The neointimal thickness and area of the experimental side were significantly reduced compared with the control side (P < 0.05). Scanning electron microscopy showed that the anastomotic intima of the experimental side was covered with one layer of intact and regular endothelium cells with deposition of little blood components, but the anastomotic intima of the control side was covered with irregular endothelium cells and deposited with a lot of blood cells and fibrins. Transmission electron microscopy showed the anastomotic intima of the control side that rich in vascular smooth muscle cells and the matrix of the intima was composed of regular collagenous fibers, and that of the experimental side consisted of several types of cells with a lot of foreign particles in the matrix., Conclusion: It is safe and effective to locally use low dose of paclitaxel carried by FG in the prevention of vascular anastomotic site intimal hyperplasia. Paclitaxel molecules can penetrate the graft wall and stay in the anastomotic intima more than four weeks postoperationally.

The sorting nexins (SNXs) constitute a large group of PX domain-containing proteins that play critical roles in protein trafficking. We report here the solution structure of human sorting nexin 22 (SNX22). Although SNX22 has <30% sequence identity with any PX domain protein of known structure, it was found to contain the alpha/beta fold and compact structural core characteristic of PX domains. Analysis of the backbone dynamics of SNX22 by NMR relaxation measurements revealed that the two walls of the ligand binding cleft undergo internal motions: on the picosecond timescale for the beta1/beta2 loop and on the micro- to millisecond timescale for the loop between the polyproline motif and helix alpha2. Regions of the SNX22 structure that differ from those of other PX domains include the loop connecting strands beta1 and beta2 and the loop connecting helices alpha1 and alpha2, which appear to be more mobile than corresponding loops in other known structures. The interaction of dibutanoyl-phosphatidylinositol-3-phosphate (dibutanoyl-PtdIns(3)P) with SNX22 was investigated by an NMR titration experiment, which identified the binding site in a basic cleft and indicated that ligand binding leads only to a local structural rearrangement as has been found with other PX domains. Because motions in the loops are damped out when dibutanoyl-PtdIns(3)P binds, entropic effects could contribute to the lower affinity of SNX22 for this ligand compared to other PX domains.

Resonance light scattering(RLS) and absorption spectra of Quercetin were studied in the present paper. In the solution with pH 3.30 to pH 6.50, the RLS singal was very strong and stable, but decreased sharply with a increase in pH when pH > 6.50. In the B-R buffer solution at pH 4.00, the RLS intensity at 497 nm was maximum and proportional to the concentration of quercetin. The linear range is 0.0-3.0 x 10(-4) mol x L(-1) with the detection limit of 3.1 x 10(-7) mol x L(-1). Quantum chemistry calculation showed that this enhancement of RLS occurred because the neutral molecules of quercetin assembled into super-molecular aggregates by 4-4' hydrogen-bond. This result completely accords with the spectral data from experiments.

Objective: To detect the relations between incidence rate of the epidemical encephalitis B and related factors, to provide a simple, valid and practical new method for forecasting encephalitis B eipdemics., Methods: Connection number between the incidence rate of encephalitis B and the historical forecast factors was computed, before ranking the first, second and the third principal factor, to remove the factor with the smallest value in the light of the connection number before comparing the newest value of forecast factors with the same kind of history while the most nearly value becoming the forecasting factor value and to establish a forecasting equation according to the factor value and the consistent degree of the incidence rate of encephalitis B at that time. Finally, to put into the new factor value to get this forecast value under this equation. Assuming that there are n' (n' >or= 2) forecast factors, this time forecast value can then be directly obtained from the average of these estimate values., Results: Using above forecast method to forecast the incidence rate of encephalitis B at certain place and year, the predicting value is very much close to the actual incidence rate. Difference between the predicting value forecasted by the above-mentioned method and the actual incidence rate is only 0.0264/100 000 with an accurate rate of 97.94%., Conclusion: This principal factor analysis forecast method based on connection number in forecasting the incidence rate of encephalitis B prevention is acceptable.

Aim: To study the pharmacokinetic characteristics of lactosaminated recombinant human growth hormone (hGH-L) in mice., Methods: The biodistribution was studied with in vivo radioactive tracer technique. The pharmacokinetics was investigated by radioimmunoassay (RIA) method of hGH-L. The results were compared with that of recombinant human growth hormone (hGH)., Results: 125I-hGH-L has remarkable livertaxis. The area under drug concentration-time curve (32686.9 microg . min . L-1) in blood and serum mean residence time (21.4 min) of hGH-L are less than that of hGH (36913.1 microg . min . L-1 and 24.9 min) (P < 0.05). In target organ liver, hGH-L distribution half life (1.8 min) and elimination half life (11.1 min) are shorter than that of hGH (2.1 min and 27.7 min) (P < 0.05). The area under drug concentration-time curve (17621.9 microg . min . L-1) of hGH-L is bigger than that of hGH(12148.2 microg . min . L-1) (P < 0.05) in liver., Conclusion: The pharmacokinetic parameters of hGH-L has obvious advantage over that of hGH.