Search

A bacterial strain YHl that used p-tert-Butylcatechol (TBC) as the sole carbon and energy source was isolated from the activated sludge of the wastewater treatment plant of a chemical factory. The strain was identified as Pseudomonas corrugate by morphological characteristics, physiological and biochemical properties, BIOLOG and the 16S rDNA sequence analysis. The degration rate of TBC was above 82% with an initial concentration of 500 mg.L-1 at 24-36°C and an initial pH of 7. 0-10. 0. Through single-factor experiments of its degradation characteristics of strain YH1, the results showed that its optimal additional carbon and nitrogen sources were sucrose and tryptone, the optimal temperature was 30°C, the optimal initial pH was 7. 0, and the optimal inoculation volume was 2%. In order to improve the TBC degradation rate, the concentrations of sucrose and tryptone and initial pH were identified as the main factors by Placket-Burman Assay. Then these factors reached their optimal region by Steepest Ascent. Finally, the optimal levels of those main factors were further optimized using Box-Behnken design and response surface analysis. The optimal conditions were as follows: sucrose concentration 3% (ρ), tryptone concentration 1. 44% (ρ), TBC concentration 400 mg.L-1, initial pH value 8. 12, inoculation amount 2. 97% (φ), temperature 30°C, training time 96 h. Under the optimal conditions mentioned above, the TBC degradation rate reached 98. 21%. Enzymology analysis and localization experiments showed that the TBC-degrading enzymes were intracellular proteins and the synthesis of catechol 1,2-dioxygenase (C12O) could be induced by TBC. Through design of specific PCR primers for the degrading enzymes, the gene encoding the catechol 1,2-dioxygenase was amplified from YHL. It was found that genes encoding TBC-degrading enzyme were located on plasmids by the plasmid detection and elimination experiments. In addition, YH1 was tolerant to high concentration of NaCl and many kinds of heavy metal ions, and resistant to multiple antibiotics. This paper provided some information for the effective treatment of complex industrial wastewater.

Objective: To study the effect of Jinlida on changes in expression of skeletal muscle lipid transport enzymes in fat-induced insulin resistance ApoE -/- mice., Method: Eight male C57BL/6J mice were selected in the normal group (NF), 40 male ApoE -/- mice were fed for 16 weeks, divided into the model group (HF), the rosiglitazone group ( LGLT), the Jinlida low-dose group (JLDL), the Jinlida medium-dose group (JLDM), the Jinlida high-dose group (JLDH) and then orally given drugs for 8 weeks. The organization free fatty acids, BCA protein concentration determination methods were used to determine the skeletal muscle FFA content. The Real-time fluorescent quantitative reverse transcription PCR ( RT-PCR) and Western blot method were adopted to determine mRNA and protein expressions of mice fatty acids transposition enzyme (FAT/CD36), carnitine palm acyltransferase 1 (CPT1), peroxide proliferators-activated receptor α( PPAR α)., Result: Jinlida could decrease fasting blood glucose (FBG), cholesterol (TC), triglyceride (TG), free fatty acid (FFA) and fasting insulin (FIns) and raise insulin sensitive index (ISI) in mice to varying degrees. It could also up-regulate mRNA and protein expressions of CPT1 and PPARα, and down-regulate mRNA and protein levels of FAT/CD36., Conclusion: Jinlida can improve fat-induced insulin resistance ApoE -/- in mice by adjusting the changes in expression of skeletal muscle lipid transport enzymes.

Triptolide is a key component of the traditional Chinese medicinal plant Thunder God Vine and has potent anticancer and immunosuppressive activities. It is an irreversible inhibitor of eukaryotic transcription through covalent modification of XPB, a subunit of the general transcription factor TFIIH. Cys342 of XPB was identified as the residue that undergoes covalent modification by the 12,13-epoxide group of triptolide. Mutation of Cys342 of XPB to threonine conferred resistance to triptolide on the mutant protein. Replacement of the endogenous wild-type XPB with the Cys342Thr mutant in a HEK293T cell line rendered it completely resistant to triptolide, thus validating XPB as the physiologically relevant target of triptolide. Together, these results deepen our understanding of the interaction between triptolide and XPB and have implications for the future development of new analogues of triptolide as leads for anticancer and immunosuppressive drugs., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Recently, a logarithmic decrease of conductivity has been observed in topological insulators at low temperatures, implying a tendency of localization of surface electrons. Here, we report quantum transport experiments on the topological insulator Bi2Te3 thin films with arrayed antidot nanostructures. With increasing density of the antidots, a systematic decrease is observed in the slope of the logarithmic temperature-dependent conductivity curves, indicating the electron-electron interaction can be tuned by the antidots. Meanwhile, the weak antilocalization effect revealed in magnetoconductivity exhibits an enhanced dominance of electron-electron interaction among decoherence mechanisms. The observation can be understood from an antidot-induced reduction of the effective dielectric constant, which controls the interactions between the surface electrons. Our results clarify the indispensable role of the electron-electron interaction in the localization of surface electrons and indicate the localization of surface electrons in an interacting topological insulator.

The realization of superconductivity at the interface between a topological insulator and an iron-chalcogenide compound is highly attractive for exploring several recent theoretical predictions involving these two new classes of materials. Here we report transport measurements on a Bi2Te3/FeTe heterostructure fabricated via van der Waals epitaxy, which demonstrate superconductivity at the interface, which is induced by the Bi2Te3 epilayer with thickness even down to one quintuple layer, though there is no clear-cut evidence that the observed superconductivity is induced by the topological surface states. The two-dimensional nature of the observed superconductivity with the highest transition temperature around 12 K was verified by the existence of a Berezinsky-Kosterlitz-Thouless transition and the diverging ratio of in-plane to out-plane upper critical field on approaching the superconducting transition temperature. With the combination of interface superconductivity and Dirac surface states of Bi2Te3, the heterostructure studied in this work provides a novel platform for realizing Majorana fermions.

Y2O3:Tb3+ and Y2O3:Tb3+, Yb3+ samples were prepared by co-precipitation method. The morphology, microstructure and fluorescence spectra at room temperature of samples were characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD) and fluorescence spectrometer, The optimal process conditions of Y2O3:Tb3+ under different doping concentrations, annealing temperature, and pH value of the solution were obtained: Tb3+ concentration is 1.5%, annealing temperature is 1400 degrees C, an alkaline solution environment, and samples under 300 nm light excitation have the largest green light emission at 543 nm. The corresponding relation of Tb3+ ion level structure and transition properties and experimental spectra were analyzed in detail, and we explained the influence mechanism of process conditions and the fluorescence quenching process mainly effects luminous intensity of samples. The energy transfer from sensitizing ions Tb3+ to active ion Yb3+ was confirmed, it made the sample have considerable emitting light in the near-infrared region; the authors described the process of cooperation conversion luminescence between the two ions from the level transition angle, and also analyzed the system of fluorescence quenching process. Test results showed that the near infrared quantum cutting can effectively improve the luminous efficiency of doped ions, and will have broad application prospects in the silicon solar cells and other fields.

Background: It is now clear that there are two histological types (type 1 and type 2) of autoimmune pancreatitis (AI P). The histological substance of type 1 AI P is known as lymphoplasmacytic sclerosing pancreatitis (LPSP) or traditional AIP, and type 2 AIP is characterized by distinct histology called idiopathic duct centric pancreatitis (IDCP). Serum IgG4 increase is considered as a marker for type 1 AI P. Far less is known about type 2 and it lacks predicting markers, so it easily leads to missed diagnosis and misdiagnosis., The Aim of This Study: The aim of this study was to describe multi-gene mutations in patients with type 2 AI P and its clinical features., Material and Methods: Three unrelated patients with type 2 AI P, 10 cases with type 1 AIP, 15 cases with other chronic pancreatitis and 120 healthy individuals were studied. The mutations and polymorphisms of 6 genes involved in chronic pancreatitis or pancreatic cancer - PRSS1, SPINK1, CFTR, MEN1, PKHD1, and mitochondrial DNA - were sequenced. Information of clinical data was collected by personal interview using a structured questionnaire., Results: Novel mutations were found in the genes encoding for MEN1 (p.546 Ala > The) and PKHD1 (c. 233586 A > G and c. 316713 C > T) from patients with type 2 AIP. What is more, the serum TCR (T cell receptor) level is relatively higher in patients with type 2 AIP than in patients with type 1 AIP and other chronic pancreatitis or normal controls. Weight loss was the major manifestation and no patients had extrapancreatic involvement in type 2 AIP., Conclusions: Type 2 AIP may occur with multi-gene mutations. For screening purposes, it is more reasonable to evaluate TCR levels in serum.

Introduction: The systematic meta-analysis of randomized controlled trials (RCTs) evaluated the effects of intraoperative ulinastatin on early-postoperative recovery in patients undergoing cardiac surgery., Methods: RCTs comparing intraoperative ulinastatin with placebo in cardiac surgery were searched through PubMed, Cochrane databases, Medline, SinoMed, and the China National Knowledge Infrastructure (1966 to May 20th, 2013). The primary endpoints included hospital mortality, postoperative complication rate, length of stay in intensive care unit, and extubation time. The physiological and biochemical parameters illustrating postoperative cardiac and pulmonary function as well as inflammation response were considered as secondary endpoints., Results: Fifteen RCTs (509 patients) met the inclusion criteria. Ulinastatin did not affect hospital mortality, postoperative complication rate, or ICU length of stay but reduced extubation time. Ulinastatin also increased the oxygenation index on postoperative day 1 and reduced the plasma level of cardiac troponin-I. Additionally, ulinastatin inhibited the increased level of tumor necrosis factor-alpha, polymorphonuclear neutrophil elastase, interleukin-6, and interleukin-8 associated with cardiac surgery., Conclusion: Ulinastatin may be of value for the inhibition of postoperative increased inflammatory agents and most likely provided pulmonary protective effects in cardiac surgery. However, larger adequately powered RCTs are required to define the clinical effect of ulinastatin on postoperative outcomes in cardiac surgery.

The aim of the present study was to optimize the pregelatinized starch technique for cell block preparation and apply this approach in cultured cells of all types of growing forms, suspension and adherent. In order to evenly mix the starch powder and the cell suspension, we crafted a special plastic dropper. To prove the effectiveness of this optimized technique we used different cell lines, NCI-H69, NCI-H345, HCT-116, SKBR3 and MDA-MB-231. The morphology features, immunocytochemistry (ICC) and fluorescent/chromogenic in-situ hybridization (FISH/CISH) on the cell block sections were evaluated. The morphology features, the ICC and ISH results of cell block sections prepared by the new method were satisfactory comparing with the results obtained in biopsies, the gold standard test for this kind of analysis. The most attractive advantage of our optimized pregelatinized starch technique is that this new method is based on cell suspensions instead of cell sediment, so with our technique every section will contain cells due to the even distribution of the starch powder and the cells forming a homogeneous cell block. To the authors' knowledge, this is the first description on cell block preparation based on cell suspension., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Objective: To investigate the treatment efficacy of tympanostomy microtube placement surgery for middle ear atelectasis., Methods: A retrospective analysis was conducted on 26 patients (28 ears) with middle ear atelectasis, who complained fullness or pressure in the ears.Otoscope showed tympanic membrane invagination, scattered or disappeared cone of light, tympanic membrane was pale and dull. The pure tone audiometry air-bone gap >10 dB. Acoustic immittance showed tympanic negative pressure. All the ears had atelectasis of I-III grade. Patients were performed tympanic membrane microtube placement under local anesthesia, and were followed up for 6-12 months., Results: Twenty-five ears recovered from the fullness after operation, in which, 23 ears reverted from type "C" to type "A" in acoustic immittance tests and the pure-tone average (PTA) of hearing thresholds were decreasing from 5 to 20 dB, while 2 ears relapse after removal of the microtube. Three ears with middle ear atelectasis of III grade were ineffectiveness. All the 26 cases had no complications including middle ear infection, tympanosclerosis, and permanent perforation after removal of the microtubes., Conclusions: The placement of tympanostomy microtube can be used to treat middle ear atelectasis, especially to the patients with middle ear atelectasis of I-II grade as it is effective on elimination of middle ear negative pressure and remission of fullness.

Vascular endothelial growth factor (VEGF) is a hypoxia-induced angiogenic protein that exhibits a broad range of biological and pathological effects in wet age-related macular degeneration and proliferative diabetic retinopathy. However, its specific mechanism is still not fully understood. Here, we examined the effects of VEGF on choroid-retinal endothelial cells (RF/6A) proliferation and tube formation, and the underlying signal pathways responsible in this process. RF/6A cells were pretreated with MEK inhibitor or PI3K inhibitor, and then incubated in a hypoxia chamber. Real-time PCR and Western blot analysis were carried out to explore VEGF expression on mRNA and protein levels. Hypoxia inducible factor-1α (HIF-1α) and VEGFR2 expression levels were also investigated in the presence and absence of hypoxic conditions. CCK-8 analysis and tube formation assay were tested under hypoxia, exogenous recombinant VEGF, and different signal pathway inhibitors, respectively. Mean while, the PI3K/Akt and MEK/ERK pathways in this process were also investigated. Our results showed that VEGF, HIF-1α, VEGFR2, p-ERK, and p-Akt were up-regulated in RF/6A cells under hypoxic conditions. MEK inhibitor (PD98059) and PI3K inhibitor (LY294002) decreased ERK and Akt activity, respectively, and reduced VEGF expression. VEGF-induced RF/6A proliferation and tube formation requires MEK/ERK and PI3K/Akt signaling, and both of the two pathways were needed in regulating VEGF expression. These suggest that VEGF plays an important role in RF/6A proliferation and tube formation, and MEK/ERK and PI3K/Akt pathway may be responsible for this process.

Glomus tumor is usually a small, benign tumor and typically occurs in the dermis or subcutis or soft tissue of the extremities and rarely in the visceral locations. Its occurrence in the main bronchus is extremely rare. The current case reported a 30-year-old woman with dyspnea on exertion and hemoptysis, she had a glomus tumor which has large size, deep location and exhibits an infiltrative margin as well as increased atypical mitotic figures. These characteristics suggest malignant behavior. However, there is little data regarding glomus tumors arising in the bronchus, the need for caution in diagnosing this case as a malignant glomus tumor must be highlighted. Therefore, the diagnosis of bronchial glomus tumor of uncertain malignant potential was favored. To the best of our knowledge, both the type and the location of this glomus tumor are extremely rare. Accumulation of more cases are needed to clarify their diagnosis and significance since there is little data regarding glomus tumors arising in the bronchus.

Aim: To describe protease serine 1 (PRSS1) gene mutations in patients with autoimmune pancreatitis (AIP) and the clinical features of AIP., Methods: Fourteen patients with AIP, 56 with other chronic pancreatitis, 254 with pancreatic cancer and 120 normal controls were studied. The mutations and polymorphisms of four genes involved with pancreatitis or pancreatic cancer, PRSS1, SPINK1, CFTR and MEN1, were sequenced. The pathogenic mechanism of AIP was investigated by comparing the wild-type expression system with the p.81Leu→Met mutant expression system., Results: Two novel mutations (p.81Leu→Met and p.91Ala→Ala) were found in PRSS1 gene from four patients with AIP. PRSS1_p.81Leu→Met mutation led to a trypsin display reduction (76.2%) combined with phenyl agarose (Ca(2+) induced failure). Moreover, the ratio of trypsin/amylase in patients with AIP was higher than in the patients with pancreatic cancer and other pancreatitis. A large number of lymphocytes and plasma cells were found in the bile ducts accompanied by hyperplasia of myofibroblasts., Conclusion: Autoimmune pancreatitis may be related to PRSS1 gene mutations.

We herein reported a primary pulmonary papillary carcinoma with colloid-like luminal content in the glandular cavity and classic nuclear features such as pseudo-inclusions, intranuclear grooves in the tumor cell nuclei and ground glass nuclei which closely mimics papillary thyroid carcinoma. Meanwhile, lymph node in the left pulmonary hilum was involved and showed similar features to the primary pulmonary papillary carcinoma. This specific histopathological presentation caused a diagnostic dilemma.The patient didn't show previous concomitant or subsequent evidence of a thyroid tumor. Immunohistochemistry further confirmed pulmonary origin and excluded a metastasis from the thyroid, as it was thyroglobulin negative, thyroid transcription factor 1 and surfactant apoprotein A positive, which was consistent with the imageology and history.Based on the above features, the diagnosis of primary pulmonary papillary carcinoma was confirmed. Understanding the existence of papillary thyroid carcinoma-like pulmonary papillary carcinoma will avoid misdiagnosis or unnecessary clinical and radiologic investigations in future.

We present a study of the surface reactivity of a Pd/Bi2Te3 thin film heterostructure. The topological surface states from Bi2Te3, being delocalized and robust owing to their topological natures, were found to act as an effective electron bath that significantly enhances the surface reactivity of palladium in the presence of two oxidizing agents, oxygen and tellurium respectively, which is consistent with a theoretical calculation. The surface reactivity of the adsorbed tellurium on this heterostructure is also intensified possibly benefitted from the effective transfer of the bath electrons. A partially inserted iron ferromagnetic layer at the interface of this heterostructure was found to play two competing roles arising from the higher-lying d-band center of the Pd/Fe bilayer and the interaction between the ferromagnetism and the surface spin texture of Bi2Te3 on the surface reactivity and their characteristics also demonstrate that the electron bath effect is long-lasting against accumulated thickness of adsorbates.

Cell block sections serve as an important diagnostic annex for cytological smears, liquid-based SurePath cytology and the Liquid-based Thin-prep Cytology Test (TCT). A variety of methods for the preparation of cell blocks are described in the literature and the techniques in cell blocks are in continuous improvement. A new technique for making cell blocks was introduced in the present study. We first used pregelatinized starch as the frame for the cell block, which is a really simple and economic method, because it can be carried out at room temperature without additional special instruments. We have performed hematoxylin and eosin (HE) staining, immunohistochemistry analysis and fluorescence in situ hybridization (FISH) in the cell block sections in 122 cytological specimens. The results demonstrated in this article show that pregelatinized starch is a useful frame for cell blocks. The pregelatinized starch can effectively collect even a few cells with powerful adhesiveness. Therefore, this new technique for making cell blocks is especially useful for cytologic samples with low cellularity, such as cerebrospinal fluid specimens.

Resistance is (not) futile: The yatakemycin biosynthetic gene cluster involves the ytkR2 gene, which encodes a protein with homology to a recently discovered bacterial DNA glycosylase. Genetic validation in vivo, biochemical assays, and in vitro mutagenesis studies revealed that YtkR2 confers resistance for the bacteria by specifically recognizing and cleaving the YTM-modified base (see scheme)., (Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background: Circular stapled hemorrhoidopexy (CSH) is an effective technique for treating prolapsing hemorrhoids; but urgency and anal stenosis are common postoperative complications. The aim of this study was to assess the efficacy and postoperative outcomes of partial stapled hemorrhoidopexy (PSH), compared with CSH., Methods: Seventy-two consecutive patients with grade III and IV hemorrhoids who met the inclusion/exclusion criteria were divided in a non-randomized manner to undergo either PSH (n = 34) or CSH (n = 38). Intraoperative and postoperative parameters in both groups were collected and compared., Results: The postoperative visual analog score for pain at first defecation was significantly lower in the PSH group than that in the CSH group (P = 0.001). Fewer patients in the PSH group experienced postoperative urgency, compared with those in the CSH group at 12 h, 1 day, and 7 days after surgery (P = 0.025, P = 0.019, and P = 0.043, respectively). Gas incontinence occurred in 3 patients (7.9%) in the CSH group, but in none of patients in the PSH group (P = 0.242). Postoperative anal stenosis developed in one patient (2.6%) in the CSH group, but in none of the patients in the PSH group (P = 1.0). The 2-year recurrence rate was 2.9 and 5.3%, respectively, in the PSH and CSH groups (P = 1.0)., Conclusions: The 2-year recurrence rate is similar in patients with grade III-IV hemorrhoids treated with PSH or CSH. However, PSH is associated with less postoperative pain, fewer episodes of urgency, and no anal incontinence or anal stenosis.

Purpose: This study was designed to assess the safety, efficacy, and postoperative outcomes of partial stapled hemorrhoidopexy (PSH)., Methods: A prospective study was conducted between February and March 2010. PSH was performed with single-window anoscopes for single isolated hemorrhoids, bi-window anoscopes for two isolated hemorrhoids, and tri-window anoscopes for three isolated hemorrhoids or circumferential hemorrhoids. The data pertaining to demographics, preoperative characteristics and postoperative outcomes were collected and analyzed., Results: Forty-four eligible patients underwent PSH. Single-window anoscopes were used in 2 patients, and bi- and tri-window anoscopes in 6 and 36 patients. The blood loss in patients with single-window, bi-window, and tri-window anoscopes was 6.0 ml (range 5.0-7.0 ml), 5.0 ml (range 5.0-6.5 ml), and 5.0 ml (4.5-14.5 ml) (P = 0.332). The mean postoperative visual analog scale score for pain was 3 (range, 1-4), 2 (range 1-4), 3 (range 2-6), 1 (range 0-3), 1 (range 0-2) and 2 (range 2-4) at 12 h, days 1, 2, 3, and 7, and at first defecation. The rate of urgency was 9.1%. No patients developed anal incontinence or stenosis. The 1-year recurrence rate of prolapsing hemorrhoids was 2.3%., Conclusions: Partial stapled hemorrhoidopexy appears to be a safe and effective technique for grade III-IV hemorrhoids. Encouragingly, PSH is associated with mild postoperative pain, few urgency episodes, and no stenosis or anal incontinence.

A series of N-((1,3-diphenyl-1H-pyrazol-4-yl)methyl)aniline derivatives (5a-8d) have been designed and synthesized, and their biological activities were also evaluated as potential antitumor and cyclin dependent kinase 2 (CDK2) inhibitors. Among all the compounds, compound 5a displayed the most potent CDK2/cyclin E inhibitory activity in vitro, with an IC(50) of 0.98 ± 0.06 μM. Antitumor assays indicated that compound 5a owned high antiproliferative activity against MCF-7 and B16-F10 cancer cell lines with IC(50) values of 1.88 ± 0.11 and 2.12 ± 0.15 μM, respectively. Docking simulation was performed to insert compound 5a into the crystal structure of CDK2 at active site to determine the probable binding model. Based on the preliminary results, compound 5a with potent inhibitory activity in tumor growth may be a potential anticancer agent., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Background: Osthole (Ost), a natural coumarin derivative, has been shown to inhibit many pro-inflammatory mediators and block voltage-gated Na+ channels. During inflammation, acidosis is an important pain inducer which activates nociceptors by gating depolarizing cationic channels, such as acid-sensing ion channel 3 (ASIC3). The aim of this study was to examine the effects of Ost on nucleus pulposus-evoked nociceptive responses and ASIC3 over-expression in the rat dorsal root ganglion, and to investigate the possible mechanism., Material/methods: Radicular pain was generated with application of nucleus pulposus (NP) to nerve root. Mechanical allodynia was evaluated using von Frey filaments with logarithmically incremental rigidity to calculate the 50% probability thresholds for mechanical paw withdrawal. ASIC3 protein expression in dorsal root ganglions (DRGs) was assessed with Western blot and immunohistochemistry. Membrane potential (MP) shift of DRG neurons induced by ASIC3-sensitive acid (pH6.5) was determined by DiBAC(4) (3) fluorescence intensity (F.I.)., Results: The NP-evoked mechanical hyperalgesia model showed allodynia for 3 weeks, and ASIC3 expression was up-regulated in DRG neurons, reaching peak on Day 7. Epidural administration of Ost induced a remarkable and prolonged antinociceptive effect, accompanied by an inhibition of over-expressed ASIC3 protein and of abnormal shift of MP. Amiloride (Ami), an antagonist of ASIC3, strengthened the antinociceptive effect of Ost., Conclusions: Up-regulation of ASIC3 expression may be associated with NP-evoked mechanical hyperalgesia. A single epidural injection of Ost decreased ASIC3 expression in DGR neurons and the pain in the NP-evoked mechanical hyperalgesia model. Osthole may be of great benefit for preventing chronic pain status often seen in lumbar disc herniation (LDH).

Yatakemycin (YTM), an antitumor natural product, represents the most potent member of a class of potent anticancer natural products including CC-1065 and duocarmycins. Herein we describe the biosynthetic gene cluster of YTM, which was identified by genome scanning of Streptomyces sp. TP-A0356. This cluster consists of 31 open reading frames (ORFs) and was localized to a 36 kb DNA segment. Moreover, its involvement in YTM biosynthesis was confirmed by cluster deletion, gene replacement, and complementation. Inactivation of ytkT, which encodes a radical S-adenosylmethionine (SAM) protein, created a mutant strain that failed to produce YTM but accumulated a new metabolite, which was structurally elucidated as a precursor that was related to the formation of the cyclopropane ring. More importantly, biochemical characterization of the radical SAM-dependent enzyme YtkT revealed that it is a novel C-methyltransferase and contributes to an advanced intermediate during formation of the cyclopropane ring through a radical mechanism in the YTM biosynthetic pathway. On the basis of in silico analysis, genetic experiments, structure elucidation of the novel intermediate, and biochemical characterization, a biosynthetic pathway for yatakemycin was proposed, which sets the stage to further investigate the novel enzymatic mechanisms and engineer the biosynthetic machinery for the production of novel analogues.

One of the most treatable causes of lower back pain and associated sciatica is lumbar disc herniation (LDH), which is characterized by rupture of the hard outer wall (annulus fibrosis) in a lumbar intervertebral disc. In the current study, we aimed to: (1) develop and characterize a rat model of sciatica induced by LDH, while introducing a novel method of epidural catheterization; (2) use this model to evaluate the effect of osthole on pain due to LDH, and (3) gain insight into the mechanisms through which osthole affects sciatica induced by LDH. The results indicate that our newly developed rat model maintained mechanical allodynia for 28 days without reduction. Moreover, cyclooxygenase-2 (COX-2) and nitric oxide synthase (NOS) were overexpressed in the associated inflammatory response, which is consistent with clinical manifestations of the disease. We then used this model to study the effect and mechanisms through which osthole affected pain due to LDH. Our study suggests that osthole is capable of reversing hyperalgesia due to LDH, potentially through modulation of activity of COX-2 and NOS, two important proteins for the exacerbation of pain due to LDH. Finally, a molecular modeling simulation showed that osthole has unique binding capabilities to both NOS and COX-2. As the model-induced mechanical hyperalgesia response was consistent, and the position of the catheter tip and the extension/spreading of the drug in the epidural space were reliable, this study developed an improved model to study remedies for sciatic pain. Moreover, our studies demonstrate that osthole may be a feasible treatment for the reduction of pain due to hyperalgesia., (Copyright © 2012 S. Karger AG, Basel.)

Objective: To study the effects of osthole on sciatica induced by lumber disc herniation and its mechanisms., Methods: 54 male SD rats were randomly divided into 6 groups. Model (M) group (n = 12): Autologous nucleus pulposus was harvested from the tail and applied to the L5 dorsal root ganglion (DRG) and epidural space. Epidural catheterization was performed. Control(C) group (n = 12): On the basis of nucleus pulposus group, 50 microL tween-80 was administered epidurally on the day 6th after surgery. T2 (n = 6), T6 (n = 12), T13 (n = 6) and T20 (n = 6) group: 50 microL 2% osthole was administered epidurally on the 2th, 6th, 13th day and 20th after surgery respectively. General behaviors were observed and 50% paw withdrawal threshold (50% PWT) was measured 1 day before surgery, on the 1st, 3th, 7th,14th, 21th, 28th day after surgery, immediately before and 1 hour after osthole or tween-80 administration in each group. On the 7th day after surgery, the left L5 DRGs were obtained for detecting the expression of NOS and COX-2 in M, C and T6 group with 6 rats., Results: No lameness or autophagy was oberserved. 50% PWT decreased after surgery (P < 0.05). In T2 and T6 group, 50% PWT after osthole administration were significantly higher than those of M group and C group (P < 0.05), which recovered to the same level as 1 day before surgery (P > 0.05). In T13 and T20 group, 50% PWT 1 hour after osthole administration were significantly higher than those of M group and C group (P < 0.05), which recovered to the same level as 1 day before surgery (P > 0.05), but on days after 1 hour after administration, there was no significant difference when 50% PWT compared with M group or nucleus C group (P > 0.05). NOS positive cells and COX-2 positive cells were no significant difference when M group compared with C group (P > 0.05). But these positive cells in T6 group were significantly lower than those of M group and C group (P < 0.05)., Conclusion: 50 microL 2% osthole could completely inhibit the mechanical allodynia in the rat model of sciatica induced by lumbar disc herniation when it was administered epidurally on 2 or 6 day after surgery. But when administered on 13 or 20 day after surgery, its analgesic effect was transient. The effect of 50 microL 2% osthole epidural administration on day 6 after surgery on the rat model of sciatica induced by lumbar disc herniation may relate to inhibition of the expression of COX-2 and NOS in DRG.

Triptolide (1) is a structurally unique diterpene triepoxide isolated from a traditional Chinese medicinal plant with anti-inflammatory, immunosuppressive, contraceptive and antitumor activities. Its molecular mechanism of action, however, has remained largely elusive to date. We report that triptolide covalently binds to human XPB (also known as ERCC3), a subunit of the transcription factor TFIIH, and inhibits its DNA-dependent ATPase activity, which leads to the inhibition of RNA polymerase II-mediated transcription and likely nucleotide excision repair. The identification of XPB as the target of triptolide accounts for the majority of the known biological activities of triptolide. These findings also suggest that triptolide can serve as a new molecular probe for studying transcription and, potentially, as a new type of anticancer agent through inhibition of the ATPase activity of XPB.

Transcription factors (TFs) are proteins that bind to specific promoter regions of their target genes and regulate gene transcription. Many of these factors have been found to influence flowering. Lycoris longituba exhibits a great deal of diversity in flower color and flower form, making it a suitable model for the study of floral development. We have identified 338 putative TFs from more than thirty thousand ESTs sequenced from the floral tissue of L. longituba, and validated them using real-time RT-PCR. Fifty-one of the TFs were recognized as being potentially flower-specific, and the expression patterns of some of them during six flowering phases have been elucidated. Homolog annotation and phylogenetic analysis revealed that some TFs that belong to several TF families, such as MADS, MYB-related, NAC, and ABI3-VP1, were suggested to play important roles in the flowering process. Our dataset may be used to identify priority target TF genes for further study., (2010 Elsevier Inc. All rights reserved.)

Objective: To evaluate the effect of Sapylin combined with intraperitoneal and systemic chemotherapy for advanced colon cancer following radical operation on local recurrence, hepatic metastasis, and overall survival rate., Methods: From Jan. 2004 to Dec. 2005,132 patients with stage II or III colon carcinoma after radical operation were randomly divided into two groups: Sapylin combined with chemotherapy(Sapylin) group and the control group. Toxic reaction, local recurrence, hepatic metastasis, and overall survival rate between two groups were compared., Results: Both groups successfully completed the trial. There was no significant difference in toxic reaction between two groups, the recurrence and hepatic metastasis rate in Sapylin group were lower than those in the control group(9/60, 15.0% vs. 22/72, 30.6%; 11/60,18.3% vs. 22/72, 34.7%, respectively), which were statistically significant (P<0.05 respectively). The 3- year survival rate in Sapylin group was higher than that in control group(73.3% vs. 54.2%), which was statistically significant (P<0.05)., Conclusion: Sapylin combined with intraperitoneal and systemic chemotherapy can effectively decrease local recurrence and hepatic metastasis,and improve the survival in patients with advanced colon cancer following radical operation.

3-(1-Arylsulfonylalkyl)indoles as electrophiles in the N-heterocyclic carbene-catalyzed umpolung reaction of aldehydes were realized for the first time. This intermolecular Stetter-type reaction features the commercially available catalyst and mild reaction conditions, providing alpha-(3-indolyl) ketone derivatives in high yields for a wide range of substrates.

Objective: To establish a magnetic particle antibody immunoassay (MPAIA) for the detection of specific antibody in sera of schistosomiasis patients., Methods: Fluorescein isothiocyanate (FITC) was used to label Schistosoma japonicum soluble egg antigen (Sj-SEA). Anti-human IgG coated with alkaline phosphatase (ALP) as enzyme-labeled second antibody, and magnetic beads were coupled with sheep anti-FITC antibody as solid phase. Phenolphthale in monophosphate was used as substrate to set up MPAIA for the detection. Serum samples from cases with schistosomiasis or other helminth infections were tested., Results: The positive rate of MPAIA was 96.7% (116/120) with the sera of S. japonicum-infected cases. No cross reaction was observed with sera of trichinellosis, paragonimiasis or cysticercosis cases. The positive titer with reference sample was 1: 1,600. The precision was lower than 10%. The MPAIA tips can be stored at 4 degrees C for 12 months., Conclusion: MPAIA shows a high sensitivity, proper specificity and long-term validity for schistosomiasis detection.

Aim: To evaluate hepatic recurrence and prognostic factors for survival in patients with surgically resected hilar cholangiocarcinoma in a single institution over the last 13 years., Methods: From 1994 to 2007, all patients with hilar cholangiocarcinoma referred to a surgical clinic were evaluated. Demographic data, tumor characteristics, and outcome were analyzed retrospectively. Outcome was compared in patients who underwent additional liver resection with resection of the tumor., Results: Of the 69 patients submitted to laparotomy for tumor resection, curative resection (R(0) resection) was performed in 40 patients, and palliative resection in 29. Thirty-one patients had only duct resection, and 38 patients had combined duct resection with liver resection including 34 total or part caudate lobes. Curative rates with the combined hepatectomy were significantly improved compared with those without additional hepatectomy (27/38 vs 13/31; chi2 = 5.94, P < 0.05). Concomitant liver resection was associated with a decreased incidence of initial recurrence in liver one year after surgery (11/38 vs 23/31; chi2 = 13.98, P < 0.01). The 3-year survival rate after R(0) resection was 30.7% and was 10.5% for palliative resection. R(0) resection improved the 3-year survival rate (30.7% vs 10.5%; chi2 = 12.47, P < 0.01)., Conclusion: Hepatectomy, especially including the caudate lobe combined with bile duct resection should be considered standard treatment to cure hilar cholangiocarcinoma.

Simultaneously isolating more than one receptor of natural product remains a challenge to chemical genetics. Using cyclosporine A as an affinity probe and an optimized phage display cloning procedure, not only cyclophilin A, but also cyclophilin B was isolated as the full-length gene clone from a human brain cDNA library. This optimized protocol can be used to select protein targets of chemicals dependent on the binding affinity rather than on the relative abundance in cells.

Background: Cholangiocarcinoma is a highly aggressive, fatal malignancy, which is resistant to all current therapeutic approaches. The recent elevation in the incidence of cholangiocarcinoma has highlighted the need for novel approaches targeting the molecular basis of its invasiveness. Previously we reconstructed a RhoC antisense eukaryotic expression vector and transfected it into a cholangiocarcinoma cell line (QBC939) by the lipofectamine method. This study was undertaken to determine the effect of the antisense RhoC gene on the proliferation and invasion capacity of QBC939., Methods: Antisense RhoC cDNA was transfected into QBC939 with lipofectin 2000. The cell growth curve was constructed to determine the proliferation rate of cells; flow cytometry was used to analyze cell cycle changes of the tumor cells; and a Boyden chamber was used to assess the invasive ability of the cells before and after gene transfection., Results: After the antisense RhoC cDNA was transfected, the number of colonies formed was significantly lower than that in the other two groups (54+/-8 vs. 91+/-11 vs. 90+/-9, P<0. 05) so was the number of the cells which crossed to the lower surface of the matrigel-coater filters (36+/-6 vs. 96+/-12 vs. 95+/-7, P<0.05). There was also a higher percentage of transfected cells in G1 phase than in the other two groups (52.5% vs. 43.4% vs. 43.7%)., Conclusion: The antisense RhoC gene can suppress the capacities of proliferation and invasion in a cholangiocarcinoma cell line in vitro.

Objective: To study the effects of catecholamines on human preadipocyte proliferation and differentiation., Methods: Catecholamines (epinephrine, isoprenaline and noradrenaline) were added to the culture media of human preadipocytes. The proliferation of cells, the expression of GPDH and lipid droplet accumulation in cytoplasm were observed and recorded. The functions of alpha and beta receptors were examined with adding alpha and beta receptor blockers., Results: Epinephrine and isoprenaline stimulated human preadipocyte proliferation and inhibited GPDH up-regulation during differentiation. The three types of catecholamines inhibited lipid accumulation in cell differentiation. The beta-adrenoceptors played a key role during the process., Conclusion: Human preadipocytes responded to catecholamine characteristically. The result would be applicable in the study of drugs for obesity.

The effects of fibroblast growth factor basic (bFGF), transforming growth factor alpha (TGF alpha), recombinant human epidermal growth factor (EGF), recombinant human tumor necrosis factor alpha (TNF alpha), and recombinant interleukin 1 alpha (IL-1 alpha) on lymphatic angiogenesis were assessed in cultured newborn bovine lymphatic endothelial cells (NBLEC). bFGF, TGF alpha, and EGF stimulated the proliferation of NBLEC in a dose-dependent manner, but the combination of either two growth factors did not show synergistic effects on NBLEC DNA synthesis. TNF alpha and IL-1 alpha suppressed the multiplication of NBLEC. Treatment with bFGF markedly increased the migration of NBLEC. The tissue plasminogen activator (t-PA) activity was enhanced by bFGF. TNF alpha also promoted NBLEC t-PA activity. These results suggest that bFGF is a major multifunctional lymphatic endothelial cell targeted cytokine, and both growth and pro-inflammatory cytokines exert differential regulatory effects on lymphatic endothelial cell proliferation, migration and t-PA activity.