Comprehensive Genomic and Transcriptomic Analysis of Sclerosing Pneumocytoma.

Sclerosing pneumocytoma is a rare and distinct lung neoplasm whose histogenesis and molecular alterations are the subject of ongoing research. Our recent study revealed that AKT1 internal tandem duplications (ITD), point mutations, and short indels were present in almost all tested sclerosing pneumocytomas, suggesting that AKT1 mutations are a major driving oncogenic event in this tumor. Although the pathogenic role of AKT1 point mutations is well established, the significance of AKT1 ITD in oncogenesis remains largely unexplored. We conducted comprehensive genomic and transcriptomic analyses of sclerosing pneumocytoma to address this knowledge gap. RNA-sequencing data from 23 tumors and whole-exome sequencing data from 44 tumors were used to obtain insights into their genetic and transcriptomic profiles. Our analysis revealed a high degree of genetic and transcriptomic similarity between tumors carrying AKT1 ITD and those with AKT1 point mutations. Mutational signature analysis revealed COSMIC signatures 1 and 5 as the prevailing signatures of sclerosing pneumocytoma, associated with the spontaneous deamination of 5-methylcytosine and an unknown etiology, respectively. RNA-sequencing data analysis revealed that the sclerosing pneumocytoma gene expression profile is characterized by activation of the PI3K/AKT/mTOR pathway, which exhibits significant similarity between tumors harboring AKT1 ITD and those with AKT1 point mutations. Notably, an upregulation of SOX9, a transcription factor known for its involvement in fetal lung development, was observed in sclerosing pneumocytoma. Specifically, SOX9 expression was prominent in the round cell component, whereas it was relatively lower in the surface cell component of the tumor. To the best of our knowledge, this is the first comprehensive investigation of the genomic and transcriptomic characteristics of sclerosing pneumocytoma. Results of the present study provide insights into the molecular attributes of sclerosing pneumocytoma and a basis for future studies of this enigmatic tumor.

[Consistency evaluation of multidimensional quality attributes of Atractylodis Macrocephalae Rhizoma by fusion of multi-source information].

The quality control of Chinese medicinal decoction pieces is one of the key tasks in the traditional Chinese medicine industry. In this study, multi-source information fusion was employed to fuse the data from near-infrared spectroscopy, electronic tongues, and other tests and establish an overall quality consistency evaluation method for Atractylodis Macrocephalae Rhizoma, which provided methodological support for the overall quality evaluation of Atractylodis Macrocephalae Rhizoma. The near-infrared spectroscopy information was measured in both static and dynamic states for 23 batches of Atractylodis Macrocephalae Rhizoma samples from different sources, and the electronic tongue sensory information, moisture content, and leachate content were measured. The overall quality of Atractylodis Macrocephalae Rhizoma was evaluated by multi-source information fusion. The results showed that the near-infrared spectroscopy information of 16122103, 801000509, 801000352, 701003656, HX21L01, and 160956 was different from that of other batches of Atractylodis Macrocephalae Rhizoma powder in the static state, and 701003298, 16122103, 701003656, 701003107, 801000229, and 18090404 were the different batches in the dynamic state. The moisture content showed no significant difference between batches. The leachate content in the batch 801000509 was different from that in other batches. The electronic tongue sensory information of 150721004, 151237, 160703004, HX21M01, HX21K04, HX21K01, and 601003516 was different from that of other batches. Furthermore, data layer fusion was employed to analyze the overall quality of Atractylodis Macrocephalae Rhizoma. Four batches, 150721004, HX21M01, HX21K04, and HX21K01, showed the parameters exceeding the 95% control limits and differed from the other samples in terms of the overall quality. This study integrated the information of moisture, near-infrared spectroscopy, and other sources to evaluate the quality consistency among 23 batches of Atractylodis Macrocephalae Rhizoma samples, which provides a reference for the quality consistency evaluation of Chinese medicinal decoction pieces.

Constant p.L424H Mutation in GTF2I in Micronodular Thymomas With Lymphoid Stroma: Evidence Supporting Close Relationship With Type A and AB Thymomas.

Micronodular thymoma with lymphoid stroma is a rare thymic neoplasm characterized by discrete nodules of epithelial tumor cells separated by abundant lymphoid stroma. The genetic features of micronodular thymoma with lymphoid stroma remain largely unexplored. Owing to the interference of abundant intratumoral, nonneoplastic lymphoid cells, a highly sensitive approach is necessary to study genetic changes in these tumors. In this study, we used a highly sensitive next-generation sequencing assay using the molecular barcoding Ion AmpliSeq HD technology to study the most commonly mutated genes in thymomas, including GTF2I, HRAS, NRAS, KRAS, and TP53. A total of 12 cases of micronodular thymomas with lymphoid stroma were tested, and 2 cases also had areas of type A thymoma in their tumor bed. Two micronodular thymic carcinomas with lymphoid stroma, a histological mimic of micronodular thymoma, were also included for comparison. Recurrent p.L424H mutations in GTF2I were found in all the cases of micronodular thymoma with lymphoid stroma but not in the cases of micronodular thymic carcinomas. In addition, 3 cases of micronodular thymoma with lymphoid stroma also had concomitant HRAS and/or KRAS mutations. Our study showed that p.L424H mutations in GTF2I is a constant genetic feature of micronodular thymoma with lymphoid stroma. This finding strongly suggests that micronodular thymoma with lymphoid stroma is closely related to type A and AB thymomas because they all share p.L424H mutations in GTF2I.

Genomic Profiling With Large-Scale Next-Generation Sequencing Panels Distinguishes Separate Primary Lung Adenocarcinomas From Intrapulmonary Metastases.

The distinction between different separate primary lung cancers (SPLCs) and intrapulmonary metastases (IPMs) is a challenging but clinically significant issue. Histopathology-based classification is the current practice; however, it is subjective and affected by interobserver variability. Recently, next-generation sequencing (NGS) panels have been used in lung cancer diagnostics. This study aimed to investigate the value of large-scale NGS panels for distinguishing between SPLCs and IPMs. A total of 32 patients with 69 lung adenocarcinomas were included. Comprehensive histopathologic assessments of multiple pulmonary adenocarcinomas were performed independently by 3 pathologists. The consensus of histopathologic classification was determined by a majority vote. Genomic analysis was performed using an amplicon-based large-scale NGS panel, targeting single-nucleotide variants and short insertions and deletions in 409 genes. Tumor pairs were classified as SPLCs or IPMs according to a predefined molecular classification algorithm. Using NGS and our molecular classification algorithm, 97.6% of the tumor pairs can be unambiguously classified as SPLCs or IPMs. The molecular classification was predictive of postoperative clinical outcomes in terms of overall survival (P = .015) and recurrence-free interval (P = .0012). There was a moderate interobserver agreement regarding histopathologic classification (κ = 0.524 at the tumor pair level). The concordance between histopathologic and molecular classification was 100% in cases where pathologists reached a complete agreement but only 53.3% where they did not. This study showed that large-scale NGS panels are a powerful modality that can help distinguish SPLCs from IPMs in patients with multiple lung adenocarcinomas and objectively provide accurate risk stratification.

[Critical quality attribute characterization of big brand TCM: taste formulation optimization strategy and application of Xiaoer Ganmao Oral Liquid based on taste quality attribute].

The taste is the quality attribute for the development and production of traditional Chinese medicine(TCM). To improve the medication compliance of the big brand TCM, Xiaoer Ganmao Oral Liquid, a correlation model between the electronic tongue sensor signal value and human sensory evaluation score was established, and an optimization strategy of taste improvement for Xiaoer Ganmao Oral Liquid was developed with the key techniques of statistical experimental design. Based on the above model, the optimal formulation was determined as follows: aspartame content of 1-2 mg·mL~(-1), acesulfame-K content of 1.5-3 mg·mL~(-1), and steviol glycoside content of 1-2 mg·mL~(-1). Furthermore, the optimal formulation was verified by human sensory evaluation. Therefore, the taste of Xiaoer Ganmao Oral Liquid was improved. Taking Xiaoer Ganmao Oral Liquid as an example, the present study developed the taste formulation optimization method based on the correlation between the electronic tongue and human sensory evaluation, which is expected to provide an important reference to improve the taste of oral liquid of TCM.

[Critical quality attribute assessment of big brand traditional Chinese medicine: study on NIR field detection method of chemical properties of moisture in Tongren Niuhuang Qingxin Pills].

For the field detection problems of critical quality attribute(CQA) of moisture content in traditional Chinese medicine(TCM) manufacturing process, big brand TCM Tongren Niuhuang Qingxin Pills were used as the carrier, to establish a moisture content NIR field detection model with or without cellophane in real world production with use of near infrared(NIR) spectroscopy combined with stoichiometry. With the moisture content determined by drying method as reference value, the partial least square method(PLS) was used to analyze the correlation between the spectrum and the moisture reference value. Then the spectral pretreatment methods were screened and optimized to further improve the accuracy and stability of the model. The results showed that the best quantitative model was developed by the spectral data pretreatment of standard normal variate(SNV) with the latent variable factor number of 2 and 7 of Tongren Niuhuang Qingxin Pills with or without cellophane samples. The prediction coefficient of determination(R_(pre)~2) and standard deviation of prediction(RMSEP) of the model with cellophane samples were 0.765 7 and 0.157 2%; R_(pre)~2 and RMSEP of the model without cellophane samples were 0.772 2 and 0.207 8%. The NIR quantitative models of moisture content of Tongren Niuhuang Qingxin Pills with and without cellophane both showed good predictive performance to realize the rapid, accurate and non-destructive quantitative analysis of moisture content in such pills, and provide a method for the field quality control of the critical chemical attributes of moisture in the manufacturing of big brand TCM.

[Critical quality attribute assessment of big brand traditional Chinese medicine: quality control of Tongren Niuhuang Qingxin Pills in texture and sensory attributes].

Texture sensory attributes are the key items in quality control of Chinese medicinal honeyed pills. The purpose of this study is to develop a quality control method for assessing the texture sensory attributes of Chinese medicinal honeyed pills based on real-world Tongren Niuhuang Qingxin pilular masses and finished products. First, parameters of texture profile analysis(TPA) were optimized through single factor and central composite design(CCD) experiments to establish a detection method for texture sensory attri-butes of Tongren Niuhuang Qingxin Pills. The results showed that the established detection method was stable and reliable, with the optimal parameters set up as follows: deformation percentage of 70%, detection speed at 30 mm·min~(-1), and interval time of 15 s. Furthermore, 540 data points yielded form six texture sensory attributes of pills from 30 batches were subjected to multivariate statistical process control(MSPC) with Hotelling T~2 and squared prediction error(SPE) control charts to establish the quality control method of Tongren Niuhuang Qingxin Pills. This study is expected to provide a reference for improving the quality control system of Chinese medicinal honeyed pills.

Epidemiology Analysis of Streptococcus pyogenes in a Hospital in Southern Taiwan by Use of the Updated emm Cluster Typing System.

emm typing is the most widely used molecular typing method for the human pathogen Streptococcus pyogenes (group A streptococcus [GAS]). emm typing is based on a small variable region of the emm gene; however, the emm cluster typing system defines GAS types according to the nearly complete sequence of the emm gene. Therefore, emm cluster typing is considered to provide more information regarding the functional and structural properties of M proteins in different emm types of GAS. In the present study, 677 isolates collected between 1994 and 2008 in a hospital in southern Taiwan were analyzed by the emm cluster typing system. emm clusters A-C4, E1, E6, and A-C3 were the most prevalent emm cluster types and accounted for 67.4% of total isolates. emm clusters A-C4 and E1 were associated with noninvasive diseases, whereas E6 was significantly associated with both invasive and noninvasive manifestations. In addition, emm clusters D4, E2, and E3 were significantly associated with invasive manifestations. Furthermore, we found that the functional properties of M protein, including low fibrinogen-binding and high IgG-binding activities, were correlated significantly with invasive manifestations. In summary, the present study provides updated epidemiological information on GAS emm cluster types in southern Taiwan.

N-Glycosylation of Human R-Spondin 1 Is Required for Efficient Secretion and Stability but Not for Its Heparin Binding Ability.

R-spondin 1 (Rspo1) plays an essential role in stem cell biology by potentiating Wnt signaling activity. Despite the fact that Rspo1 holds therapeutic potential for a number of diseases, its biogenesis is not fully elucidated. All Rspo proteins feature two amino-terminal furin-like repeats, which are responsible for Wnt signal potentiation, and a thrombospondin type 1 (TSR1) domain that can provide affinity towards heparan sulfate proteoglycans. Using chemical inhibitors, deglycosylase and site-directed mutagenesis, we found that human Rspo1 and Rspo3 are both N-glycosylated at N137, a site near the C-terminus of the furin repeat 2 domain, and Rspo2 is N-glycosylated at N160, a position near the N-terminus of TSR1 domain. Elimination of N-glycosylation at these sites affects their accumulation in media but have no effect on the ability towards heparin. Introduction of the N-glycosylation site to Rspo2 mutant at the position homologous to N137 in Rspo1 restored full glycosylation and rescued the accumulation defect of nonglycosylated Rspo2 mutant in media. Similar effect can be observed in the N137 Rspo1 or Rspo3 mutant engineered with Rspo2 N-glycosylation site. The results highlight the importance of N-glycosylation at these two positions in efficient folding and secretion of Rspo family. Finally, we further showed that human Rspo1 is subjected to endoplasmic reticulum (ER) quality control in N-glycan-dependent manner. While N-glycan of Rspo1 plays a role in its intracellular stability, it had little effect on secreted Rspo1. Our findings provide evidence for the critical role of N-glycosylation in the biogenesis of Rspo1.

Tandem Duplication and Random Loss for mitogenome rearrangement in Symphurus (Teleost: Pleuronectiformes).

BACKGROUND: The mitochondrial genomes (mitogenomes) of flatfishes (Pleuronectiformes) exhibit highly diversified types of large-scale gene rearrangements. We have reported that the mitogenomes of Crossorhombus azureus (Bothidae), Samariscus latus (Samaridae) and Cynoglossus fishes (Cynoglossidae) show different types of gene rearrangements. RESULTS: In the present study, the complete mitogenomes of two Symphurus species (Cynoglossidae), Symphurus plagiusa and Symphurus orientalis, were determined. The gene order in the S. plagiusa mitogenome is the same as that of a typical vertebrate (without any gene rearrangements). Surprisingly, large-scale gene rearrangements have occurred in S. orientalis. In the rearranged fragment from the control region (CR) to the WANCY tRNA cluster (tRNA cluster of tRNA-W, tRNA-A, tRNA-N, tRNA-C and tRNA-Y) in the S. orientalis mitogenome, tRNA-V and tRNA-M have been translocated to the 3' end of the 16S rRNA gene, with six large intergenic spacers over 20 bp in length. In addition, an origin for light-strand replication (OL) structure that is typically located in the WANCY region was absent in both the S. plagiusa and S. orientalis mitogenomes. It is generally recognized that a sequence in the WANCY region that encodes tRNAs forms a hairpin structure (OL-like structure) and can act as the OL when the typical locus is lost. Moreover, an additional OL-like structure was identified near the control region in the S. plagiusa mitogenome. CONCLUSIONS: The positions of the intergenic spacers and the rearranged genes of the S. orientalis mitogenome strongly indicate that the mechanism underlying the rearrangement of this mitogenome was Tandem Duplication and Random Loss. Additionally, two OL-like regions substituting for the typical locus were found in the S. plagiusa mitogenome. We speculate that the ancestral mitogenomes of S. plagiusa and S. orientalis also had this characteristic, such that if both OL-like structures functioned during mitochondrial replication, they could initiate duplicate replications of the light strand (L-strand), leading to duplication of the region between the two structures. We consider that this mechanism may account for the gene duplication that occurred during the gene rearrangement process in the evolution of the ancestral mitogenome to the S. orientalis mitogenome.

[Collisional relaxation between highly vibrationally excited Na2 and Ar and H2].

Energy transfer rate constants were measured for excited rovibrational levels of Na2 (X1Σ(g)+). Stimulated emission pumping was used to excited the levels v = 33-51, J = 11 via A-X transition. Laser induced fluorescence was used to follow the collision dynamics. Energy transfer processes induced by collisions with Ar and H2 were investigated. The decay curves for the parent level populations gave good fits to single exponential function. At v = 33-51, the total transfer rate constants increase linearly with vibrational quantum number. Parameterized expressions for the (48, 11)--to--(47, J) rate constants were fitted to the fractional populations of the satellite lines. This produced sets of relative rate constants. Absolute rate constants were then obtained by normalizing the sums of the relative rate constants to the total removal rate constants. For Na2 (v) + Ar, no multiquantum vibrational transfer was detected. For Na2 (v) + H2, a significant fraction of the initial population of highly vibrationally excited Na2 (X v = 48) relaxes to lower vibrational level (Δ v = -5). The time scale is much shorter than the known collisional lifetimes of the intervening vibrational levels and thus a sequential single-quantum relaxation mechanism can be explicitly ruled out. For v = 48, at least 40% of the initially prepared population, undergoes multiquantum vibrational relaxation. We discuss possible explanations of this result.

[Time resolved distribution of excitation energy in collisions of vibrationally excited KH with CO2].

The vibrational levels of KH(X1 sigma+ v" = 0-3) were generated in the reaction of K(5P) with H2. The vibrationally excited KH(v" = 17) was populated by an overtone pump-probe configuration Different characteristics of collisional energy transfer in highly and lowly excited vibrational levels of KH and CO2 were investigated through measuring the time-resolved distribution of vibrational energy in KH(v" = 17.3) + CO2 collisions. For KH(v" = 17), there existed three principal regions of vibration temperature (T(v)) in this equilibration process. The initial phase consists of very rapid fall in T(v) within - 5 micros, and the vibrational energy of KH(v" = 17) is mainly transferred to the vibrational levels of CO2 (00 degrees 1) or high rotational levels of CO2 (00 degrees 0). The second phase (5-20 micros) has a slight decline in T(v), and the process of energy transfer to vibrational levels or high rotational levels of CO2 has already finished. The vibration temperature of the third phase has a slightly more rapid decline compared with the last phase. This phase shows that the process of transfer to lowly rotational levels and translation energy of CO2 is accelerated. The equilibration of vibrationally excited KH (v" = 3) in CO2 was also investigated. There are similarities to the behavior of KH (v" = 17) in CO2 plot, but also are significant differences. Once the initial resonant V-R exchange has equalized vibrational temperatures, there is a very slow linear decline in T(v) with equilibrium attained within -80 micros. This same point is reached within 15 micros for KH (v" = 17). The data demonstrate that single rate coefficient measurements are unlikely to capture the complex nature of processes that generally are multistaged with different relaxation rates characterizing each different stage. Examination of the quantum state distributions reveals that these distinct stages reflect the dominance of specific energy transfer mechanisms, some of which are inherently fast and others are much slower. The energy gain into CO2 resulting from collisions with excited KH was probed using transient absorption techniques. Distributions of nascent CO2 rotational populations in both ground (00 degrees 0) state and the vibrationally excited (00 degrees 1) state were determined. A kinetic model was developed to describe rate coefficients for appearance of CO2 states resulting from collisions with excited KH. These experiments show that collisions resulting in CO2 (00 degrees 0) are accompanied by substantial excitation in rotation while the vibrationally excited CO2 (00 degrees 1) state has rotational energy distributions near the initial distributions.

[Transfer energy disposal in collisions of NaK (6(1)sigma+) with H2].

The radiative lifetimes and rate coefficients for deactivation of high lying 6(1)sigma+ state of NaK by collisions with H2 were studied. An OPO laser was set to a particular 2(1)sigma+ <-- 1(1)sigma+ transition. Another single mode Ti sapphire laser was then used to excite molecule from 2(1)sigma+ level to the 6(1)sigma+ state. The predissociation was monitored by the atomic potassium emission at the 3D --> 4P (1.7 microm) or the S --> 4P (1.24 microm), while bound state radiative processes were monitored by total fluorescence from the upper state to the various levels, all studied as a function of H2 density. The values for predissociation, collisional dissociation and collisional depopulation rate coefficients were obtained. The decay signal of the time resolved fluorescence from the 6(1)sigma+ --> 2(1)sigma+, 6(1)sigma+ -->1(1)sigma+ or 2(1)sigma+ --> 1(1)sigma+ transition was monitored. Based on the Stern-Volmer equation, the radiative lifetimes were monitored for 6(1)sigma+ --> 2(1)sigma+ and 2(1)sigma+ --> 1(1)sigma+ transition. The rate coefficients for deactivation of collisions with H2 were monitored for 6(1)sigma+ --> 2(1)sigma+, 6(1)sigma+ --> 1(1)sigma+ and 2(1)sigma+ -->1(1)sigma+. When the density of H2 was 10(19) cm(-3), the total collisional transfer energy (15 426 cm(-1)) and radiative energy (10 215 cm(-1)) were obtained. The relative fraction ((f(v)), (f(R)), (f(T)) of average energy disposal was derived as (0.58, 0.03, 0.39); (f(v)), (f(R)), (f(T)) represent separately the relative fraction of average energy disposal among vibration, rotation and translation. The major vibrational and translational energy release supports the assumption that the 6(1)sigma(+) -H2 collision occurs primarily in a collisional energy transfer mechanism. In this experiment, alkali molecules relative energy population ratio was determined through using the time integrated intensity, so we can get the total transfer energy. That the NaK (6(1)sigma+) energy transfers to the H2 vibrational, rotational and translational energy was quantitatively given for the first time, which illustrates the collisional mechanism.

[Role of PI3K/Akt pathway in effect of paeoniflorin against Aß25-35-induced PC12 cell injury].

OBJECTIVE: To study the role of PI3K/Akt pathway in the neuroprotective effect of paeoniflorin on PC12 cells. METHOD: The paeoniflorin group (5, 10, 20 µmol · L(-1)) was pretreated for 30 min, and then added with Aß25-35 (20 µmol · L(-1)) for interaction for 24 h. Inhibitor LY294002 (10 µmol · L(-1)) was pretreated for 30 min before the action of paeoniflorin (10 µmol · L(-1)). The MTT colorimetric method was used to detect the cell viability. The apoptosis rate was tested by the FITC-Annexin V/PI staining. The protein expression of p-AKT, Bax, Bcl-2 and cleaved caspase-3 protein were detected by Western blot analysis. RESULT: Paeoniflorin could significantly inhibit the Aß25-35-induced PC12 cell toxicity and apoptosis. Its protection effect may be achieved by up- regulating AKT phosphorylation level, increasing Bcl-2 protein expression, reducing Bax protein expression, inhibiting the activation of caspase-3. Inhibitor LY294002 could weaken the above protective effects of paeoniflorin. CONCLUSION: Paeoniflorin could activate PI3K/Akt signaling pathway to protect the PC12 cell injury induced by Aß25-35.

Complete mitogenome sequences of four flatfishes (Pleuronectiformes) reveal a novel gene arrangement of L-strand coding genes.

BACKGROUND: Few mitochondrial gene rearrangements are found in vertebrates and large-scale changes in these genomes occur even less frequently. It is difficult, therefore, to propose a mechanism to account for observed changes in mitogenome structure. Mitochondrial gene rearrangements are usually explained by the recombination model or tandem duplication and random loss model. RESULTS: In this study, the complete mitochondrial genomes of four flatfishes, Crossorhombus azureus (blue flounder), Grammatobothus krempfi, Pleuronichthys cornutus, and Platichthys stellatus were determined. A striking finding is that eight genes in the C. azureus mitogenome are located in a novel position, differing from that of available vertebrate mitogenomes. Specifically, the ND6 and seven tRNA genes (the Q, A, C, Y, S1, E, P genes) encoded by the L-strand have been translocated to a position between tRNA-T and tRNA-F though the original order of the genes is maintained. CONCLUSIONS: These special features are used to suggest a mechanism for C. azureus mitogenome rearrangement. First, a dimeric molecule was formed by two monomers linked head-to-tail, then one of the two sets of promoters lost function and the genes controlled by the disabled promoters became pseudogenes, non-coding sequences, and even were lost from the genome. This study provides a new gene-rearrangement model that accounts for the events of gene-rearrangement in a vertebrate mitogenome.

Re-evaluation of the pathogenic roles of nonstructural protein 1 and its antibodies during dengue virus infection.

Dengue virus (DENV) infection can cause life-threatening dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). Vascular leakage and abnormal hemorrhage are the two major pathogenic changes found in these patients. From previous studies, it is known that both antibodies and cytokines induced in response to DENV infection are involved in the immunopathogenesis of DHF/DSS. However, the role of viral factors during DENV infection remains unclear. Nonstructural protein 1 (NS1), which is secreted in the sera of patients, is a useful diagnostic marker for acute DENV infection. Nevertheless, the roles of NS1 and its antibodies in the pathogenesis of DHF/DSS are unclear. The focus of this review is to evaluate the possible contributions of NS1 and the antibodies it induces to vascular leakage and abnormal hemorrhage during DENV infection, which may provide clues to better understanding the pathogenesis of DHF/DSS.

[Protective effects of paeoniflorin against aBETA25-35-induced oxidative stress in PC12 cells].

OBJECTIVE: To investigate the neuroprotective effects and mechanism of paeoniflorin on Abeta25-35-induced oxidative stress in PC12 cells. METHOD: The proliferation of induced PC12 cells were investigated by the MTT method. The leakage rate of lactate dehydrogenase (LDH), the intracellular content of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were also measured. The changes of the intracellular reactive oxygen species (ROS) level and the mitochondrial membrane potential (MMP) were investigated by flow cytometry using DCFH-DA and Rh123 staining. The protein expression of HO-1, Cyt C and cleaved Caspase-3 was detected by Western blot. RESULT: The results showed that paeoniflorin at different doses (5, 10, 20 micromol . L-1) could increase the cell viability and activities of antioxidant enzyme (GSH-Px, SOD and HO-1), prevent LDH release and lipid peroxidation (MDA) production, decrease the level of intracellular ROS, increase MMP, inhibit the release of cytochrome c from the mitochondria and attenuate activation of Caspase-3. CONCLUSION: Paeoniflorin can protect PC12 cells from Abeta25-35 oxidative stress injuries. The mechanism may be related to its antioxidant action and inhibition of mitochondria-mediated caspase signaling pathways.

[Rovibrational state distributions of H2 in collisional energy transfer between NaK (6(1)sigma(+)) and H2].

The electronic to rovibrational energy transfer between the high-lying 6(1)sigma(+) state of NaK and H2 was investigated. The 6(1)sigma(+) state was excited using the optical-optical double resonance technique. Radiative processes were monitored by fluorescence from the 6(1)sigma(+) state to the ground 1(1)sigma(+) state. The CARS (coherent anti-Stokes Raman scattering) spectral technique was used to probe the internal state distribution of collisionally-populated H2 molecules. The scanned CARS spectra reveal that during E-V, R energy transfer processes H2 molecules are produced at v = 1, 2 and 3 levels. The semilog plot of the time-resolved H2 (3, 1) CARS intensity was obtained at a pressure of 4 x 10(4) Pa of H2. The slop yields an effective quenching rate of 8.09 x 10(5) s(-1). Analogically, for (3, 2), (3, 3) and (3, 5) levels, the quenching rates are 6.11 x 10(5), 4.32 x 10(5) and 2.45 x 10(5) s(-1), respectively. For (3, 1), (3, 2), (3, 3) and (3, 5) levels, the population ratios were obtained from scanned CARS spectral peaks. For (1, 1), (2, 1), (2, 2) and (2,3) levels, from scanned CARS spectral peaks two possible population ratios were yielded. Through shape simulations of time resolved CARS profiles under a kinetic model the actual population ratios were determined. The n2/n1, n3/n1, n4/n1, n5/n1, n6/n1, n7/n1 and n8/n1 are 0.51, 0.97, 0.45, 0.18, 0.10, 0.26 and 0.31, where n1, n2, n3, n4, n5, n6, n7 and n8 represent the number densities of H2 at (3, 1), (2, 1), (1, 1), (3, 3), (2, 3), (2, 2), (3, 2) and (3, 5) levels, respectively. The H2 molecules produced by energy transfer process were populated by 26% at the v = 1 level, 21% at v = 2 and 53% at v = 3. The major vibrational energy [(9.0 +/- 2.7) x 10(3) cm(-1)] release and the minor rotational energy [(3.9 +/- 1.1) x 10(2) cm(-1)] release are shown. Based on the Stern-Volmer equation, the radiative lifetime is (5.0 +/- 1.3) ns for the 6(1)sigma(+) state, the total rate coefficient for deactivation of 6(1)sigma(+) state by means of collisions with H2 is (2.1 +/- 0.4) x 10(-10) cm3 x s(-1). From actual population ratios the rate coefficients (in units of 10(-11) cm3 x s(-1)) for 6(1) sigma(+) -(1, 1), (2, 1), (2, 2), (2, 3), (3, 1), (3, 2), (3, 3) and (3, 5) are 5.4 +/- 1.6, 2.8 +/- 0.8, 0.6 +/- 0.2, 1.0 +/- 0.3, 5.6 +/- 1.7, 1.4 +/- 0.4, 2.5 +/- 0.8 and 1.7 +/- 0.5, respectively.

[Vibrational to rotational energy transfer between CsH (Chi1 Sigma+, nu > or = 15) and CO2].

The quenching of highly vibrational excited CsH through collisions with a 500 K bath of CO2 was investigated using the laser spectroscopy technique. CsH was formed by the Cs(7P)+H2 reaction. The pulse laser prepared CsH in the highly vibrational levels. Laser induced fluorescence was used to detect collisionally relaxed CsH. The relaxation rate coefficient of CsH (nu" = 21) with CO2 is 10 times larger than that of CsH(nu" = 15). Relaxation of CsH(nu") with H2 was also investigated. The mass effect on the collisional relaxation rate coefficients is strong. The observed collisional relaxation rate coefficients of H2 are bigger than those of CO2. Energy gain into CO2 resulting from collisions with excited CsH was probed using laser overtone spectroscopy technique. Distributions of nascent CO2 rotational population in the ground (00(0)0) state were determined. For CsH excited at nu" = 15, the scattered CO2 molecules have a rotational temperature of T(rot) = (605 +/- 50) K. For excitation at nu" = 21, the CO2 rotational temperature is T(rot) = (780 +/- 70) K. Based on the rotational temperatures, the average change in the CO2 rotational energy has a stronger dependence on the CsH initial energy. Using the ambient cell temperature, nu = 21 -2. 7 nu" = 15 was found. The nascent distributions of recoil velocities for collisions were determined from stimulated absorption line profiles of individual CO2 rotational states. For nu" = 15, scattered CO2 molecules with J = 36-48 have center of mass translational energy of = 600-972 cm(-1). For nu" = 21, the values increase to = 972-1 351 cm(-1). Based on propensity rules for collisions that favor small changes in energy and angular momentum, it is reasonable that low-J CO2 states will have lower translational energy than the high-J states. Extrapolating nu" = 15 and 21 data to the initial relative translational energy of E0 = 520 cm(-1) gives an estimate of the threshold states J(th) = 34 and 24, respectively. The onset of large is likely to occur at J states that are larger than J(th).

Establishment of normal reference range for thromboelastography based on 17,708 cases in Beijing, China.

Background: Thromboelastography (TEG) can dynamically evaluate the interaction between platelets and coagulation cascade and the effects of other cellular components on the activities of plasma factors, to comprehensively analyze the whole process of blood coagulation and dissolution. Due to differences in both the incidence rates and the coagulation state of related diseases, many studies have highlighted the necessity of establishing the normal reference ranges for TEG for local regions. The aim of the present study was to determine the local normal reference ranges according to the TEG results of 17,708 volunteers in Beijing, to explore the coagulation characteristics related to the age and sex of the study population. Methods: Reference ranges of reaction time (R), coagulation time (K), coagulation angle (α-angle), and maximum thrombus consistency (MA) for TEG in healthy adults in Beijing were determined in the physical examination of 17,708 Beijing volunteers (5,319 women and 12,389 men). The volunteers were divided into the elderly group (≥60 years old) and young and middle-aged group (20-59 years old), and the reference ranges of each group were calculated according to sex. Results: Based on the TEG results of the 17,708 volunteers who underwent physical examination, the 95% reference ranges of R, K, α-angle, and MA for TEG in Beijing were 5.1-10 min, 1.3-3.8 min, 44.9-70.2°, and 50.4-71 min, respectively. The results of R, K, α-angle, and MA for TEG between the young and middle-aged group and the elderly group, as well as between women and men were significantly different (P<0.001). Finally, reference ranges for TEG in the young and middle-aged group and the elderly group were obtained. Conclusions: Compared with the reference standards provided by previous reagent manufacturers, the coagulation factor and fibrinogen function of TEG tend to be hypocoagulable in Beijing population. We found that the young and middle-aged group had lower coagulation activity than the elderly group, and women had higher coagulation activity than men in the same group.