Role of L-Arginine on the Expression of Coagulation Factor VIII Gene.

Objective: To explore the key sites in which L-arginine affects the expression of human coagulation factor VIII gene, and to create new drug targets for the treatment of hemophilia. Methods: A total of 5 human FVIII genes (A1, A2, A3, C1 and C2) with B domain deletion were transfected into human umbilical vein endothelial cells (HUVECs) as promoters. Run-on assay and ELISA analysis were performed to observe the driving effect of each domain gene on chloramphenicol acetyl transferase (CAT) gene transcription and expression, and the effect of L-arginine on each promoter. Results: In co-culture with L-arginine, transcriptional expression of the CAT gene was not detected in the PCAT3-Basic group (negative control without promoters), PA3-CAT3-Enhancer group or PC1-CAT3-Enhancer group. The transcriptional expression of CAT gene in the PCAT3-Control group (positive control with promoters) and PA1-CAT3-Enhancer group was unchanged compared with the non-L-arginine intervention, while the transcriptional expression of CAT gene in the PA2-CAT3-Enhancer group was significantly enhanced. Conclusions: A1 and A2 domain genes had promoter function and could initiate the transcription and expression of CAT gene, but A3, C1 and C2 domain genes could not. Moreover, L-arginine can significantly enhance transcription and expression of human coagulation factor VIII via A2 domain.

Investigation on abnormal gene loci of a Chinese pedigree with hereditary combined deficiency of blood coagulation factor XI, XII, and protein S.

OBJECTIVE: In order to clarify the interaction mechanism, the phenotype and abnormal gene loci of FXI, FXII, and PS were investigated in this study. METHODS: Chinese pedigree with hereditary combined deficiency of coagulation factor (F) XI, FXII, and PS was enrolled in our study. Activated partial thromboplastin time (APTT), partial thromboplastin time (PT), FXI:C, FXII:C, and protein S (PS):C were determined using the one-stage coagulation method. FXI:antigen (Ag), FXII:Ag, and PS:Ag were detected using enzyme-linked immunosorbent assay (ELISA). Exons and introns of the FXI, FXII, and PS genes were amplified by polymerase chain reaction (PCR), and gene sequencing results were analyzed using Chromas software. RESULTS: A deletion of two bases located in introns A-149 and-150 within the FXI gene of the proband, his father, wife, and both sons. A missense variant in exon 14 (GGT â†’ AGT, Gly542Ser) within FXII of the proband, his parents, and both sons. Four variants in exon 4 within the PS gene of all members of the pedigree: GTT â†’ GTG (Val46Val), CGC â†’ CTC (Arg49Leu), CGT â†’ CAT (Arg60His), and CAG â†’ TAG (Gln61stop). CONCLUSIONS: None of the pedigree members showed a tendency for bleeding or thrombosis. Therefore, we speculated that the lack of coagulation factors counteracted the lack of PS, restoring the balance between the coagulation and anticoagulation systems. Another possible explanation is that these defects individually have only partial penetrance.

Role of Ca2+, Calnexin and Calreticulin in Platelet from Adult Patients with Chronic Immune Thrombocytopenic Purpura.

BACKGROUND: Adult chronic immune thrombocytopenia (chronic ITP) is a common autoimmune hemorrhagic disease characterized by decreased platelet production and increased platelet destruction, leading to thrombocytopenia. In this study, Ca2+, calnexin (CNX) and calreticulin (CRT) within platelets from adult patients with chronic ITP were investigated. METHODS: Platelets were isolated from blood specimen collected from 20 adult patients with chronic ITP and 20 healthy volunteers. Ca2+, CNX and CRT were determined by flow cytometry, and the results were analyzed with EXPO32 ADC software. RESULTS: Flow cytometry showed the expressions of Ca2+ (74.19±19.40% vs 22.79±10.47%) was elevated (P<0.05). However, CNX (15.10±7.32% vs 41.79±14.45%) and CRT (25.11±12.66% vs 38.58±12.02%) were decreased markedly in platelets from adult patients with chronic ITP (P<0.05 compared with healthy volunteers). CONCLUSION: Based on enhanced expression of Ca2+ and attenuated expression of CNX and CRT in patients with chronic ITP, Ca2+ concentration and its associated down-regulated proteins may be important regulatory signals in the pathogenesis of chronic ITP.

[Prediction of crown leaf mass per area for young Larix olgensis plantation]. / äººå·¥é•¿ç™½è½å¶æ¾å¹¼é¾„æž—æ ‘å† æ¯”å¶é‡.

Leaf mass per area (LMA) is an important parameter in the construction of the ecosystem process models. Accurate prediction of the dynamic validation of canopy LMA is of significance to improve the accuracy of ecosystem process models. We conducted vertical whorl-by-whorl sampling and analyzed LMA in different seasons for Larix olgensis plantation in Maoershan in Shangzhi, Heilongjiang Province, China. We analyzed the vertical and developmental variations of LMA and their main effective factors, established the dynamic prediction model of LMA for young L. olgensis plantation. The results showed that the LMA decreased with the increases of relative depth into crown (RDINC) in the vertical direction of the crown. The range of LMA in the vertical direction after leaf expanded was significantly larger than that during leaf expanding. During the different development periods of leaves, LMAs increased first and then remained stable, and this trend gra-dually weakened with the increases of crown depth. The Ra2 values were lower than 0.6 when RDINC or DOY were used as the single variable to model LMA, but were increased by 0.19 when both of them being used, and the model performed well in validation (ME=0.54 g·m-2, MAE=5.74 g·m-2). LMA varied across different crown whorls and different leaf development periods. The LMA model constructed with RDINC and DOY could well describe the vertical and temporal variations of LMA. The simulation of crown LMA provided a basis for clarifying crown development and a foundation for the establishment of ecological process model.

The diagnostic profile of rare acute myelomegakaryoblastic leukemia.

Acute myelomagakaryocytic leukemia is a diagnostic and therapeutic challenge owing to its heterogeneity and overlapping features with other types of acute leukemia. In order to build a diagnostic profile, we analyzed the biological, clinical and hematologic characteristics of acute myelomagakaryocytic leukemia. We found that, in three patients diagnosed with acute myelomagakaryocytic leukemia, there were two types of leukemia cells. One type was myeloblastic with positive peroxidase (POX) stainig and the expression of antigens CD13 and CD33. The other type was megakaryoblastic with negative POX staining and the expression of antigens CD36, CD41, CD42a and CD61. Three patients displayed the same cytogenetic abnormality, a (9: 22) translocation. Among the three patients with RT-PCR, two patients displayed BCR-ABL fusion gene amplification and one patient showed a previously undescribed OTT-MAL fusion gene amplification.

Apoptosis in platelets from adult patients with chronic idiopathic thrombocytopenic purpura.

Adult chronic idiopathic thrombocytopenic purpura (cITP) is a chronic and usually life-long haemorrhagic disorder in which enhanced platelet destruction and weakened platelet production lead to thrombocytopenia. Platelets were isolated from blood samples collected from 40 adult patients with cITP and 40 healthy volunteers. Mitochondrial membrane potential (ΔΨm) and plasma membrane phosphatidylserine externalization were determined by flow cytometry, and activation of caspase-3 and expressions of Bax, Bak and Bcl-xL were analysed by western blotting. Flow cytometry showed increased mitochondrial depolarization and lower ΔΨm in platelets from adult patients with cITP. In addition, plasma membrane phosphatidylserine externalization was observed on platelets from adult patients with cITP, but rarely from healthy volunteers. Western blot analysis of platelet proteins revealed that, in adult cITP patients, caspase-3 was activated, which cleaved gelsolin and to release a 47-kDa fragment. Moreover, the expressions of Bax and Bak were elevated, and Bcl-xL was decreased markedly in platelets from adult patients with cITP. Our findings reveal, based on loss of mitochondrial membrane potential (Δψm), phosphatidylserine exposure, caspase-3 activation, enhanced expression of Bax and Bak, and attenuated expression of Bcl-xL, that platelet death in the pathogenesis of thrombocytopenia in chronic ITP in adults is apoptotic.

[Changes of light response indicators and its impact factors of Larix principis-rupprechtii on Saihanba Forest Farm, China.] / å¡žç½•ååŽåŒ—è½å¶æ¾é’ˆå¶å ‰å“åº”æŒ‡æ ‡å˜åŒ–è§„å¾‹åŠå ¶å½±å“å› ç´ .

We examined the differences of maximum light use efficiency (LUEmax), most effective light intensity (PARe), light compensation point (LCP) and light saturation point (LSP) in diffe-rent vertical positions and different phenological periods of tree crown in Larix principis-rupprechtii plantation in Saihanba Mechanical Forest Farm of Hebei Province. We analyzed the main influencing factors for all the variables. The results showed that LUEmax increased with the increasing crown depth, and that PARe, LCP and LSP decreased with the increasing crown depth. Such a result indicated that upper crown had higher utilization efficiency to strong light and that the lower crown was more efficient to adapt to weak light environment. During the leaf development in the growth period, the LUEmax approximately increased except in July, while the changes of other photosynthe-tic-light factors showed a tendency of unimodal curve. Environmental factors were significantly correlated with the indices of light response, mainly due to the stress reaction of leaf stomata to surrounding environment. Understanding the spatial and seasonal changes of the photosynthetic-light indicators was important for formulating scientific management measures, optimizing stand structure, improving local microenvironment and maximizing forest productivity.

Investigation on glucocorticoid receptors within platelets from adult patients with immune thrombocytopenia.

Objective: The expression of glucocorticoid receptors within platelets from newly diagnosed Immune Thrombocytopenia (ITP) patients in the adult was investigated.Methods: GR expression in platelets from newly diagnosed ITP patients and healthy controls was measured using flow cytometry. Subsequently, platelets RNA and proteins were isolated and used for confirming the flow cytometry results by using RT-qPCR and ELISA.Results: Flow cytometry showed that the percentages of platelets expressing GRα and GRß from ITP patients were significantly higher than those from healthy controls (P < 0.05). qPCR and ELISA confirmed that GRα and GRß were increased at both RNA transcription and protein expression levels within platelets from ITP patients compared with healthy controls.Conclusion: We speculated that the up-regulation of glucocorticoid receptor within platelets may be an important biological feature of platelets in patients with ITP, and may also play an important role in the treatment of ITP, which is worthy of further study.

From Supramolecular Species to Self-Templated Porous Carbon and Metal-Doped Carbon for Oxygen Reduction Reaction Catalysts.

The preparation of carbon materials usually involves the decomposition of precursors and the reorganization of the as-generated fragments. However, the cleavage of bonds and the simultaneous formation of new bonds at nearly the same positions prevents effective yet precise fabrication. Herein, a supramolecular precursor, cucurbit[6]uril, that contains multiple bonds with distinct bond strengths is proposed to decouple the twin problem of simultaneous bond cleavage and formation, allowing multistage transformations to hierarchical porous carbon and metal-doped carbon in a single yet effective pyrolysis step without the need of a template or additional purification. As a proof-of-concept, the Fe-doped carbon electrocatalysts realized a Pt/C-like half-wave potential of 0.869 V vs. RHE and small Tafel slope of 51.3 mV dec-1 in oxygen reduction reaction.

Activating Inert Metallic Compounds for High-Rate Lithium-Sulfur Batteries Through In Situ Etching of Extrinsic Metal.

Surface reactions constitute the foundation of various energy conversion/storage technologies, such as the lithium-sulfur (Li-S) batteries. To expedite surface reactions for high-rate battery applications demands in-depth understanding of reaction kinetics and rational catalyst design. Now an in situ extrinsic-metal etching strategy is used to activate an inert monometal nitride of hexagonal Ni3 N through iron-incorporated cubic Ni3 FeN. In situ etched Ni3 FeN regulates polysulfide-involving surface reactions at high rates. Electron microscopy was used to unveil the mechanism of in situ catalyst transformation. The Li-S batteries modified with Ni3 FeN exhibited superb rate capability, remarkable cycling stability at a high sulfur loading of 4.8 mg cm-2 , and lean-electrolyte operability. This work opens up the exploration of multimetallic alloys and compounds as kinetic regulators for high-rate Li-S batteries and also elucidates catalytic surface reactions and the role of defect chemistry.

Study on the Role of Calreticulin Within Platelet from Adult Patients with Chronic Immune Thrombocytopenic Purpura.

To observe the differences in proteins between adult patients with chronic immune thrombocytopenic purpura (ITP) and healthy adults. 30 patients with chronic ITP and 30 healthy controls were enrolled into the study. The platelet total protein was extracted from peripheral venous blood of 10 chronic ITP patients and 10 healthy controls respectively, and subjected to two-dimensional electrophoresis (2-DE) to find the differential protein spot between chronic ITP patients and healthy controls, then the differential protein spots were identified by mass spectrometry. Subsequently, platelets RNA and proteins were isolated from the other 20 chronic ITP patients and 20 healthy controls respectively, and used for confirming the 2-DE and mass spectrometry results by using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and enzyme linked immunosorbent assay (ELISA). 2-DE combined with mass spectrometry revealed that calreticulin (CRT) expressed normally within platelets from healthy controls, while it reduced within platelets from patients with chronic ITP. qPCR and ELISA confirmed that CRT was decreased at both RNA transcription and protein expression levels within platelets from chronic ITP patients compared with healthy controls. Decreased transcription and expression of CRT within platelets may play an important role in the pathogenesis of chronic ITP, which is worthy of further study.

[Effect of Aerobic/Phosphorus Granules on Start-up of Partial Nitrification Granular Sludge].

The different effects of additional aerobic granules (AGs) and phosphorus removal granules (PRGs) on the start-up and stable operation of partial nitrification granular sludge (PNGS) were compared at room temperature(22-28℃). The results showed that in the first stage (days 0-22), partial nitrification was accomplished on day 19 for the three reactors (R1, R2, and R3). In the second stage (days 23-56), 20% AGs and 20% PRGs were added to R2 and R3 to induce PNGS. The start-up of the granules of the three reactors was successfully achieved. The mean particle sizes of R1, R2, and R3 reached 412 µm at day 76, 468 µm at day 42, and 400 µm at day 56. In the third stage (days 57-108), because the influent ammonia load increased from 0.4 kg·(m3·d)-1 to 0.5 kg·(m3·d)-1 and the COD load increased from 0.2 kg·(m3·d)-1 to 0.5 kg·(m3·d)-1, the mean particle sizes of R1 and R2 increased significantly. The average particle sizes of R1 and R2 reached 689 µm and 893 µm by the end of the operation (day 108), but sludge expansion occurred in R3. The inoculation of either AGs or PRGs could quickly achieve granulation, but the PNGS inoculated with the AGs could adapt to higher C/N and be more tolerable to shock loads and long-term stable operation.

Preliminary investigation about the expression of tubulin in platelets from patients with iron deficiency anemia and thrombocytosis.

OBJECTIVE: In order to inquire into the pathogenesis of increased platelet counts in peripheral blood of patients with iron deficiency anemia (IDA), the phenomenon of thrombocytosis was confirmed, and then the expression of tubulin within platelets from IDA patients was investigated. METHODS: Peripheral blood samples were collected from 79 patients with IDA and were divided into 2 groups, group of IDA with normal platelet counts (34 cases), and group of IDA with increased platelet counts (thrombocytosis) (45 cases). Additionally, 45 peripheral blood samples from healthy volunteers were enrolled as a group of healthy controls. Count of platelets in peripheral blood was detected by means of LH-780 hematology analyzer and hemocytometer under a microscope respectively, and analyzed statistically. RESULTS: There was no statistical difference between platelet counts detected by LH-780 hematology analyzer and hemocytometer under a microscope (P > .05). The mean fluorescence intensity (MFI) of both α-tubulin and ß-tubulin within platelets from IDA patients with thrombocytosis was significantly less than that from healthy volunteers and IDA patients with normal platelet counts (P < .01), and there was no statistical difference between the latter two groups (P > .05). CONCLUSION: Some patients with IDA are accompanied by thrombocytosis, from which the expression of α-tubulin and ß-tubulin within platelets reduced obviously compared with those with normal platelet counts and healthy controls respectively. It is implied that downregulation of tubulin probably is a part of the pathogenesis leading to increased platelet counts in IDA.

[Effect of Seeding Single/Mixed Sludge on Rapid Start-up of an ANAMMOX Reactor].

The experiment explored the effect of different seeding sludge on the rapid start-up of an anaerobic ammonium oxidation (ANAMMOX) reactor by seeding a single type of denitrified granular sludge and a mixed sludge composed of denitrified granular sludge and aerobic nitrification sludge (the volume ratio of the mixed sludge was 2:1) in two sequencing batch reactors (SBR), R0 and R1, respectively. The results indicated that R0 was started up successfully on day 64 with a nitrogen removal rate (NRR) of 0.26 kg·(m3·d)-1, while R1 was started up by day 47 with a NRR of 0.30 kg·(m3·d)-1, which was shorter than R0 by 17 d. In the enrichment stage, reddish sludge appeared in R1, and the characteristics of anaerobic ammonium oxidation of the system were more obvious than in R0. After the reactor was started up successfully, the stoichiometric ratio of R0 was 1.20 and 0.34, respectively, and the stoichiometric ratio of R1 was 1.26 and 0.21, which was closer to the theoretical values of 1.32 and 0.26. The mixed liquor suspended solids (MLSS) and mixed liquor volatile suspended solids (MLVSS) of R0 were restored to 51% (4.2 g·L-1) and 38% (2.3 g·L-1)of the initial seeding sludge, respectively, while the MLSS and MLVSS of R1 were restored to 54% (4.4 g·L-1) and 42% (2.6 g·L-1),which was higher than R0. It can be speculated that the proliferation rate of AnAOB in R1 was faster than in R0. Seeding mixed sludge can accelerate the start-up process of anaerobic ammonium oxidation with more stable N-removal performance.

[Manganese Oxidation Characteristics and Oxidation Mechanism of a Manganese-Oxidizing Bacterium Arthrobacter sp. HW-16].

Manganese is a common inorganic pollutant, which is difficult to remove from the environment. In this research, a high efficient manganese-oxidizing bacterium Arthrobacter sp. HW-16 was isolated from the manganese-rich soil using selective media. Besides, high-throughput sequencing revealed that there were significant differences of the microbial community compositions when bacteria were acclimated in different conditions, and Arthrobacte was the dominant genus in Mn(Ⅱ) containing media. In this paper, the microbiological properties of strain HW-16 and Mn(Ⅱ) oxidation mechanism were investigated. The results indicated that the maximal Mn(Ⅱ) tolerance mass concentration of strain HW-16 was 5000 mg·L-1,and it exhibited a decent Mn(Ⅱ) oxidation efficiency with the highest value of 66.28% at a Mn(Ⅱ) concentration of 3000 mg·L-1. Single factor experiments demonstrated that environmental factors could affect the growth and Mn(Ⅱ) oxidation efficiency of strain HW-16. At 30℃ or pH 7.0, at 1% or 3% salinity, and at 200 r·min-1, strain HW-16 got the highest biomass. While the highest Mn(Ⅱ) oxidation efficiency occurred at high temperature (≥40℃), high pH (≥7), high shaking speed and low salinity. Strain HW-16 could oxidize Mn(Ⅱ) by producing Mn(Ⅱ) oxidizing active factor and turn Mn(Ⅱ) into precipitation by synthesizing alkaline metabolites.

A Cooperative Interface for Highly Efficient Lithium-Sulfur Batteries.

A cooperative interface constructed by "lithiophilic" nitrogen-doped graphene frameworks and "sulfiphilic" nickel-iron layered double hydroxides (LDH@NG) is proposed to synergistically afford bifunctional Li and S binding to polysulfides, suppression of polysulfide shuttles, and electrocatalytic activity toward formation of lithium sulfides for high-performance lithium-sulfur batteries. LDH@NG enables high rate capability, long lifespan, and efficient stabilization of both sulfur and lithium electrodes.

Enhanced Electrochemical Kinetics on Conductive Polar Mediators for Lithium-Sulfur Batteries.

Lithium-sulfur (Li-S) batteries have been recognized as promising substitutes for current energy-storage technologies owing to their exceptional advantage in energy density. The main challenge in developing highly efficient and long-life Li-S batteries is simultaneously suppressing the shuttle effect and improving the redox kinetics. Polar host materials have desirable chemisorptive properties to localize the mobile polysulfide intermediates; however, the role of their electrical conductivity in the redox kinetics of subsequent electrochemical reactions is not fully understood. Conductive polar titanium carbides (TiC) are shown to increase the intrinsic activity towards liquid-liquid polysulfide interconversion and liquid-solid precipitation of lithium sulfides more than non-polar carbon and semiconducting titanium dioxides. The enhanced electrochemical kinetics on a polar conductor guided the design of novel hybrid host materials of TiC nanoparticles grown within a porous graphene framework (TiC@G). With a high sulfur loading of 3.5 mg cm-2 , the TiC@G/sulfur composite cathode exhibited a substantially enhanced electrochemical performance.

Antisense oligodeoxynucleotide against tissue factor inhibits human umbilical vein endothelial cells injury induced by anoxia-reoxygenation.

AIMS: The role of antisense oligodeoxynucleotide against tissue factor (aODN/TF) in cultured human umbilical vein endothelial cells (HUVECs) subjected to anoxia-reoxygenation (A/R) was investigated. METHODS: HUVECs were divided randomly into control group, A/R group, aODN/TF+A/R group, sense oligodeoxynucleotide (sODN/TF) + A/R group and mismatched oligodeoxynucleotide (mODN/TF) + A/R group, in the latter 3 groups, HUVECs were transfected with aODN/TF, sODN/TF and mODN/TF respectively. HUVECs in all A/R groups underwent 3 hrs of anoxia and followed by 2 hrs of reoxygenation. In order to investigate the potential mechanisms of how increased TF may contribute to A/R injury in HUVECs, another set of HUVECs were incubated with human recombinant active site blocked factor VII (FVIIai) during A/R. RESULTS: After A/R, TF expression at both mRNA and protein level was increased, furthermore, cell viability and the concentrations of SOD, GSH-PX and NO were declined, while LDH, MDA and ET-1 were overproduced (P<0.05 to 0.001 versus control group). In HUVECs of aODN/TF+A/R group, however, TF expression was inhibited, while the declined cell viability and the concentrations of SOD, GSH-PX, NO as well as the enhanced LDH, MDA and ET-1 levels occurred during A/R were ameliorated and reversed effectively (P<0.05 to 0.01 versus those in other A/R groups). The results also showed that ROS was increased and PAR-1, PAR-2, p38 MAP kinase and p42/44 MAP kinase were all activated after A/R (P<0.001 versus HUVECs under normoxia), while FVIIai inhibited the increment of ROS, PAR-1, PAR-2, p38 MAP kinase and p42/44 MAP kinase, and improved the changes of TF:C, MDA, SOD, GSH-PX, cell viability and LDH occurred during A/R (P<0.05 to 0.001 versus HUVECs without FVIIai treatment). CONCLUSION: Tissue factor plays an important role in the development of HUVECs injury induced by anoxia-reoxygenation, inhibition of TF with antisense oligodeoxynucleotide is an effective approach to ameliorate the damage.

Stachydrine, a major constituent of the Chinese herb leonurus heterophyllus sweet, ameliorates human umbilical vein endothelial cells injury induced by anoxia-reoxygenation.

Stachydrine is a major constituent of Chinese herb leonurus heterophyllus sweet, which is used in clinics to promote blood circulation and dispel blood stasis. Our study aimed to investigate the role of stachydrine in human umbilical vein endothelial cells (HUVECs) injury induced by anoxia-reoxygenation. Cultured HUVECs were divided randomly into control group, anoxia-reoxygenation (A/R) group and 4 A/R+stachydrine groups. HUVECs in the control group were exposed to normoxia for 5 hours, while in all A/R groups, HUVECs underwent 3 hours anoxia followed by 2 hours reoxygenation, and HUVECs in the 4 A/R+stachydrine groups were treated with 10(-8) M, 10(-7) M, 10(-6) M and 10(-5) M (final concentration) of stachydrine respectively. After anoxia-reoxygenation, tissue factor (TF) was over-expressed, cell viability and the concentrations of SOD, GSH-PX and NO were declined, while LDH, MDA and ET-1 were over-produced (p < 0.05 to 0.001 vs. the control group). However, in stachydrine treated groups, TF expression was inhibited at both mRNA and protein levels, while the declined cell viability and SOD, GSH-PX, NO as well as the enhanced LDH, MDA and ET-1 levels occurred during anoxia-reoxygenation were ameliorated and reversed effectively (p < 0.05 to 0.01 versus A/R group). Consequently, our findings indicate that TF plays an important role in the development of anoxia-reoxygenation injury of HUVECs, stachydrine ameliorates HUVECs injury induced by anoxia-reoxygenation and its putative mechanisms are related to inhibition of TF expression.