THIRD bedside ultrasound protocol for rapid diagnosis of undifferentiated shock: a prospective observational study.

INTRODUCTION: It is clinically challenging to differentiate the pathophysiological types of shock in emergency situations. Here, we evaluated the ability of a novel bedside ultrasound protocol (Tamponade/tension pneumothorax, Heart, Inferior vena cava, Respiratory system, Deep venous thrombosis/aorta dissection [THIRD]) to predict types of shock in the emergency department. METHODS: An emergency physician performed the THIRD protocol on all patients with shock who were admitted to the emergency department. All patients were closely followed to determine their final clinical diagnoses. The kappa index, sensitivity, specificity, positive predictive value, and negative predictive value were calculated for the initial diagnostic impression provided by the THIRD protocol, compared with each patient's final diagnosis. RESULTS: In total, 112 patients were enrolled in this study. The kappa index between initial impression and final diagnosis was 0.81 (95% confidence interval=0.73-0.89; P<0.001). For hypovolaemic, cardiogenic, distributive, and obstructive types of shock, the sensitivities of the THIRD protocol were 100%, 100%, 93%, and 100%, respectively; the sensitivity for a 'mixed' shock aetiology was 86%. The negative predictive value of the THIRD protocol for all five types of shock was ≥96%. CONCLUSION: Initial diagnostic judgements determined using the THIRD protocol showed favourable agreement with the final diagnosis in patients who presented with undifferentiated shock. The THIRD protocol has great potential for use as a bedside approach that can guide the rapid management of undifferentiated shock in emergency settings, particularly for patients with obstructive, hypovolaemic, or cardiogenic shock.

[Meta analysis of surgical treatment for old Monteggia fracture in children].

Objective: To analysis the treatment effect of the ulnar osteotomy and ring-shaped ligament reconstruction for the treatment of old Monteggia fracture in children by using Meta analyze, and the difference of clinical curative effect was compared in order to provide the basis for the selection of clinical treatment options for old Monteggia fractures in children. Methods: We searched databases such as CNKI, Wanfang database, Medline, PubMed, Embase and Science through computer, at the same time, the references of relevant documents were retrieved manually, and the data processing was carried out by the RevMan5.3 statistical software provided by the Cochrane cooperation network by incorporating the exclusion criteria. The results were obtained and analyzed. Results: A total of 17 standard literature, 438 cases, 224 cases of ulnar osteotomy, 214 cases with ring ligament repair and reconstruction were obtained. The operation scheme, which was mainly based on the lengthening of ulna osteotomy, was superior to the reconstruction of ring ligament reconstruction. The complications and second operation rates of the former was less than that of the latter. Conclusion: The surgical methods for the reconstruction of the ulna osteotomy and the ring-shaped ligament have advantages and disadvantages. The choice of the operative plan for the old Monteggia fracture should be based on the following factors: the time of the old Monteggia fracture formation, the degree of the ulnar and radial deformity and the familiarity of the operative method.

Loss of JAM-C leads to impaired esophageal innervations and megaesophagus in mice.

Megaesophagus is a disease where peristalsis fails to occur properly and esophagus is enlarged. The etiology and mechanism of megaesophagus are not well understood. In this study, we reported that junctional adhesion molecule C (JAM-C) knockout mice on a C57/B6 background developed progressive megaesophagus from embryonic day (E) 15.5 onward with complete penetrance. JAM-C knockout mice exhibited a significant reduction in the number of nerve fibers/ganglia in the wall of the esophagus. However, histological analysis revealed that the esophageal wall thickness and structure of JAM-C knockout mice at embryonic stages and young adult were comparable to that of control littermates. Thus, megaesophagus observed in JAM-C knockout mice could be attributed, at least in part, to impaired esophageal innervations. Our data suggest JAM-C as a potential candidate gene for human megaesophagus, and JAM-C knockout mice might serve as a model for the study of human megaesophagus.

293FT is a highly suitable mammalian cell line for the in vitro enzymatic activity analysis of typical P450 proteins.

Mammalian cells have been widely used for the in vitro evaluation of the functional effect of allelic variants of cytochrome P450 (CYP). The aim of this study was to determine the most suitable mammalian cell line for the in vitro drug metabolism analysis of CYP variants. Three reported cell lines (COS-7, HepG2, 293T) and one fast-growing variant of the 293 cell line 293FT were transfected with vectors expressing green fluorescent protein or typical variants of CYP2C9, CYP2C19 or CYP2D6 to investigate the protein expression levels and the catalytic activity of expressed CYP allelic variants. The transfected 293FT cells had the highest protein expression level and exhibited the highest enzymatic activity, while HepG2 cells showed the lowest activity among the four tested cell lines. Simultaneously, 293FT cells still maintained the similar relative enzymatic ratio among three typical CYP2C9 variants to that of the commonly used COS-7 cells. In addition, 293FT cells could also be used for the in vitro functional evaluation of two other typical P450 proteins, CYP2C19 and CYP2D6. Therefore, the 293FT cell line is more suitable for the in vitro enzymatic activity analysis of typical P450 proteins than any other reported mammalian cell lines.

CYP2C9 polymorphism analysis in Han Chinese populations: building the largest allele frequency database.

Genetic polymorphisms of CYP2C9 significantly influence the pharmacokinetics and pharmacodynamics of some drugs, which might result in adverse drug effects and therapeutic failure. Several studies have been performed on CYP2C9 genetic polymorphisms in Han Chinese populations. However, these studies only focused on two commonly investigated alleles, *2 and *3, in relatively small sample sizes. To scale up the gene-scanning region and determine relatively precise data on the genetic distribution pattern in Chinese populations, unrelated healthy Han Chinese volunteers from Zhejiang Province (n=1127) and Hebei (n=1000) Province were recruited as subjects for the direct sequencing of all exons of CYP2C9. As a result, 14 previously reported alleles were detected in this work, and 8 of these alleles (*14, *16, *19, *23, *27, *29, *33 and *34) were described for the first time in Chinese populations. In addition, 37 novel mutations were also detected, of which 22 variants were non-synonymous, and 21 new alleles, *36-*56, were designated by the Human CYP Allele Nomenclature Committee. In vitro functional analysis of these 22 novel CYP2C9 variants revealed that 17 mutations had a significant influence on the protein's catalytic activity. Our study provides the most accurate data on CYP2C9 polymorphisms in Han Chinese populations and detects the largest number of novel allelic variants existing to date. These new alleles will greatly enrich the current knowledge of naturally occurring CYP2C9 variants in Chinese populations.

Inhibitory effect of components from Streptomyces species on alpha-glucosidase and alpha-amilase of different origin.

The search for the effective and safe a-glucosidase and alpha-amylase inhibitors from Actinomycetaceae being antidiabetic agents is actual problem. Twenty one Streptomyces spp. of soil samples collected from different places of China were screened for the ability to produce this kind of inhibitory activities. Fermentation broth of isolated strains had absorbance between 350-190 nm. The Streptomyces strains PW003, ZG636, and ZG731 were characterized by special absorption at 280, 275, and 400 nm, respectively. Ten of the collected actinomycete strains had the ability to inhibit alpha-glucosidase or/and alpha-amylase and the fermentation broth of the same strain had inhibitory activity varied greatly depending on the enzyme source. In the process to screen the leading compounds used as antidiabetic agents, human alpha-glucosidase and alpha-amylase were revealed as the best used in trail compared with the same enzymes from other sources. Active alpha-glucosidase inhibitor was isolated from Streptomyces strain PW638 fermentation broth and identified as acarviostatin 103 by MS and N MR spectrometry. Its IC50 value was 1.25 and 12.23 microg/mL against human intestinal N-terminal maltase-glucoamylase and human pancreatic alpha-amylase, respectively.

Draft genome sequence of Streptomyces coelicoflavus ZG0656 reveals the putative biosynthetic gene cluster of acarviostatin family α-amylase inhibitors.

AIMS: The aims of this study are to obtain the draft genome sequence of Streptomyces coelicoflavus ZG0656, which produces novel acarviostatin family α-amylase inhibitors, and then to reveal the putative acarviostatin-related gene cluster and the biosynthetic pathway. METHODS AND RESULTS: The draft genome sequence of S. coelicoflavus ZG0656 was generated using a shotgun approach employing a combination of 454 and Solexa sequencing technologies. Genome analysis revealed a putative gene cluster for acarviostatin biosynthesis, termed sct-cluster. The cluster contains 13 acarviostatin synthetic genes, six transporter genes, four starch degrading or transglycosylation enzyme genes and two regulator genes. On the basis of bioinformatic analysis, we proposed a putative biosynthetic pathway of acarviostatins. The intracellular steps produce a structural core, acarviostatin I00-7-P, and the extracellular assemblies lead to diverse acarviostatin end products. CONCLUSIONS: The draft genome sequence of S. coelicoflavus ZG0656 revealed the putative biosynthetic gene cluster of acarviostatins and a putative pathway of acarviostatin production. SIGNIFICANCE AND IMPACT OF THE STUDY: To our knowledge, S. coelicoflavus ZG0656 is the first strain in this species for which a genome sequence has been reported. The analysis of sct-cluster provided important insights into the biosynthesis of acarviostatins. This work will be a platform for producing novel variants and yield improvement.

Direct mass measurements of short-lived A=2Z-1 nuclides (63)Ge, (65)As, (67)Se, and (71)Kr and their impact on nucleosynthesis in the rp process.

Mass excesses of short-lived A=2Z-1 nuclei (63)Ge, (65)As, (67)Se, and (71)Kr have been directly measured to be -46,921(37), -46,937(85), -46,580(67), and -46,320(141) keV, respectively. The deduced proton separation energy of -90(85) keV for (65)As shows that this nucleus is only slightly proton unbound. X-ray burst model calculations with the new mass excess of (65)As suggest that the majority of the reaction flow passes through (64)Ge via proton capture, indicating that (64)Ge is not a significant rp-process waiting point.

Taxonomy of the Streptomyces strain ZG0656 that produces acarviostatin alpha-amylase inhibitors and analysis of their effects on blood glucose levels in mammalian systems.

AIMS: To clarify the taxonomic status of strain ZG0656 and analyse the effects of its acarviostatin products on blood glucose levels in mammalian systems. METHODS AND RESULTS: Our program to screen for new alpha-amylase inhibitors led to the isolation of strain ZG0656. The polyphasic taxonomic study revealed that strain ZG0656 represents a novel variation of Streptomyces coelicoflavus, for which we propose the name S. coelicoflavus var. nankaiensis. Four chemically distinct alpha-amylase inhibitors, acarviostatins I03, II03, III03 and IV03, were isolated from strain ZG0656. Acarviostatins III03 and IV03 are both novel oligomers. All four acarviostatins are mixed noncompetitive porcine pancreas alpha-amylase inhibitors. Acarviostatin III03 is the most potent alpha-amylase inhibitor known to date. Moreover, in the in vitro and in vivo experiments, acarviostatins III03 showed significant inhibition of starch hydrolysis and glucose transfer to blood. CONCLUSIONS: Strain ZG0656 is a novel variation of S. coelicoflavus, whose products are novel effective alpha-amylase inhibitors. Among the products, acarviostatins III03 could significantly depress blood glucose levels in mammalian systems and be developed towards a possible therapeutic agent for diabetes. SIGNIFICANCE AND IMPACT OF THE STUDY: Acarviostatin III03 is the most potent alpha-amylase inhibitor known to date. The oligomer will benefit the research on the relationship between alpha-amylase and various inhibitors and will offer more choices in diabetes treatments.

Dexamethasone ameliorates H2S-induced acute lung injury by increasing claudin-5 expression via the PI3K pathway.

Acute lung injury (ALI) is a major outcome of exposure to high levels of hydrogen sulfide (H2S). Dexamethasone (DXM) has been used to treat ALI. However, the mechanisms involved in H2S-induced ALI and the protective mechanisms of DXM in treating ALI are still nebulous. To explore the mechanisms involved, we evaluated the role of claudin-5 in the protective effect of DXM against H2S-induced ALI. Sprague-Dawley rats were exposed to H2S to establish the ALI model. In parallel with the animal model, a cell model was also established by incubating human umbilical vein endothelial cells (HUVECs) with NaHS. Lung hematoxylin-eosin staining, electron microscope assay, and wet/dry ratio were used to identify whether the ALI was successfully induced by H2S, and changes in claudin-5 expression were detected in both rats and HUVECs. Our results revealed that claudin-5 was markedly decreased after H2S exposure and that DXM significantly attenuated the H2S-induced downregulation of claudin-5 in both rats and HUVECs. In the animal experiment, p-Akt and p-FoxO1 presented a similar tendency as claudin-5, but their levels decreased 6 h prior to the levels of claudin-5. In a further investigation, the DXM-induced protective effect on ALI and rescue effect on downregulation of claudin-5 were both blocked by LY294002. The current study demonstrated that claudin-5 was involved in the development of H2S-induced ALI and that DXM exerted protective effects through increasing claudin-5 expression by activating the phosphatidylinositol 3-kinase pathway. Therefore, claudin-5 might represent a novel pharmacological target for treating ALI induced by H2S and other hazardous gases.

[Choice of genetic model on Meta-analysis of genetic association studies: introduction of genetic model-free approach for Bayesian analysis].

Meta-analysis used for genetic association studies became popular among researchers, with the amount of published papers increased rapidly. In this paper, we will focus on the introduction on the selection of genetic models. Traditionally, methods used for Meta-analysis on genetic association studies was to calculate the statistics based on available genetic models which not only increasing the probability of false-positives but also making the interpretation of results more difficult. Hence, a critical step in the Meta-analysis of genetic association studies was to choose the appropriate inheritance model. The aim of this paper was to introduce the theory of Bayesian analysis regarding the genetic model-free approach, in performing the Meta-analysis for studies related to genetic associations.

JMJD3 promotes SAHF formation in senescent WI38 cells by triggering an interplay between demethylation and phosphorylation of RB protein.

Primary human fibroblasts undergoing oncogene-induced or replicative senescence are known to form senescence-associated heterochromatin foci (SAHF), which can stabilize the state of senescence. The retinoblastoma (RB) protein has an important role in SAHF; cells that lack active RB pathway fail to form SAHF. It has been known that the posttranslational modifications of RB, for example, phosphorylation, regulate its function. To date, whether methylation of RB impacts on the SAHF formation is unknown. Here we report that JMJD3, a histone demethylase catalyzing the tri-methylation of H3K27 (H3K27me3), can demethylate the non-histone protein RB at the lysine810 residue (K810), which is a target of the methyltransferase Set7/9. We detected a significant upregulation of JMJD3 during cellular senescence and SAHF formation in WI38 cells induced by H-RasV(12), and we found that ectopic expression of JMJD3 promoted cellular senescence and SAHF formation in WI38 cells. Furthermore, during the process of SAHF assembly, JMJD3 was transported to the cytoplasm and interacted with RB through its demethylase domain JmjC. Significantly, our data demonstrated that the JMJD3-mediated demethylation of RB at K810 impeded the interaction of RB with the protein kinase CDK4 and resulted in reduced level of phosphorylation of RB at Serine807/811 (S807/811), implicating an important role of the interplay between the demethylation and phosphorylation of RB in SAHF assembly. This study highlights the role of JMJD3 as a novel inducer of SAHF formation through demethylating RB and provides new insights into the mechanisms of cellular senescence and SAHF assembly.

Fatigue life improvement of nitrogen-ion-implanted pedicle screws.

An experimental study of the fatigue life of pedicle screws and their nitrogen-ion-implanted counterparts was conducted. Nitrogen-ion implantation parameters were varied in the first part of the study to obtain the best implantation characteristics. Using this optimal parameter for N(+)-implantation, the performance of two matched groups were compared at two bending moments near the "knee" of the S-N curves. At 3.96 N-m and 5.09 N-m of applied bending moments, the increase in the mean fatigue life for the former was 98% and for the latter 20%. The 98% prolongation was statistically significant at P less than 0.005; however, the 20% increase was statistically insignificant (P less than 0.1). The implantation depth is about 0.1 mu in the near surface of the screw. This appears to be thick enough to inhibit crack initiation at 3.96 N-m, but only marginally at 5.09 N-m. Increasing the implantation depth has the potential of increasing the fatigue life at the higher bending moments.

[Measurement of molecular weight of dextran by low angle laser light scattering photometer].

The molecular weight of two dextran standards imported from Sweden of known molecular weight and one batch of Chinese made dextran 70 were determined by low angle laser scattering photometer. The resulting values of the standards were in good agreement with the labelled values. The determined results of samples were high in precision (r = 0.95). Calculated results of the least square method were approximately the same as those of the plotting method. LALLS conditions: Solvent: pure water (n25D = 1.3314); filter: 0.22 microns VG(Millipore Corp.): Annulus = 6 degrees-7 degrees; field stop: 0.2; T = 25 degrees C.

[Immune function of rheumatoid arthritis treated by medicated-bath therapy in Tibetan medicine].

The changes of the immune function of rheumatoid arthritis before and after the Tibetan medicated-bath was observed. It showed a higher level of the rheumatoid factor (RF) titre, immunoglobulin (Ig) G, M, A and CD4 cells, but the CD8 cells was obviously lower before the treatment. Clinical data indicated that the medicated-bath had significant effective rate. In order to elucidate the mechanism of the medicated-bath upon rheumatoid arthritis the RF titre, Ig level, complement C3, 3H-TdR incorporated with lymphocytes transformation and CD3, CD4, CD8 cell level were assayed. Results showed that RF titre decreased after the bath and the negative transforming rate reached 70.6%, Ig level obviously dropped as well as the number of CD4 cells while CD8 cell level increased. The transforming stimulation index of lymphocyte cells obviously decreased. All of the above mentioned showed that there was a higher concentration of the enhancing factor of interleukin-2 (IL2-EF) involved in lymphocyte culture of rheumatoid arthritis patients. They suggested that the Tibetan medicated-bath had an immunomodulating effect on rheumatoid arthritis patients through increasing the level of CD8 cells and reducing CD4 cells.

Designing of the massive gas injection valve for the joint Texas experimental tokamak.

In order to mitigate the negative effects of the plasma disruption a massive gas injection (MGI) valve is designed for the joint Texas experimental tokamak. The MGI valve is based on the eddy-current repulsion mechanism. It has a fueling volume of 30 ml. The piston of the MGI valve is made by non-ferromagnetic material, so it can be installed close to the vacuum vessel which has a strong toroidal magnetic field. A diode is use to prevent current oscillation in the discharge circuit. The drive coil of the valve is installed outside the gas chamber. The opening characteristics and the gas flow of the MGI valve have been tested by a 60 l vacuum chamber. Owing to the large electromagnetic force the reaction time of the valve is shorter than 0.3 ms. Duration for the opening of the MGI valve is in the order of 10 ms.