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Abstract [Objective] The study aims to explore the changes of morphological, physiological, and biochemical indexes during the development of Notopterygium incisum seeds, and to provide theoretical basis for the introduction and domestication of N . incisum and the cultivation of high-quality germplasm resources. [Methods] The morphological, physiological, and biochemical indexes of N . incisum seeds at different developmental stages were determined by observation samplings and the frozen section method. [Results] During the development of N . incisum seeds, seed size and wing thickness were first increased and then decreased, while wing width was increased. The color of the outer seed coat was changed from bright blue-green to yellow-brown. The contents of starch, soluble sugar, and soluble protein were increased. Amylase activity was fluctuated in a small range; succinate dehydrogenase activity was increased; glucose-6-phosphate dehydrogenase activity was increased and decreased alternately; pyruvate kinase activity showed “M”-shape changes. POD activity was decreased while SOD activity was decreased. GA3, IAA, and ZR contents were decreased, while ABA contents were increased. The morphological indexes of the seeds were negatively correlated with the activity of SOD. The content of the storage materials was negatively correlated with the seed thickness, but positively correlated with the width of the fruit wings. [Conclusion] When N . incisum embryo is developed to pre-cardioid embryo, the outer seed coat is changed from blue-green to yellow-brown, the content of storage material is increased, and the activity of key enzymes is fluctuated and decreased. The content of growth-promoting hormones is fluctuated and decreased, while the content of stress hormones is increased.

Shengli Oilfield has abundant oil and gas resources， with multiple types of reservoirs， and logging evaluation plays an essential role in exploration and development. The development history and the characteristics of logging evaluation technologies for reservoirs were systematically reviewed to celebrate the institute's 60th birthday. Summarizing experience and predicting the future have positive significance for the development and application in the professional logging field. A series of mature evaluation technologies formed in three types of reservoirs， including conventional sandstone reservoirs， complex reservoirs， and unconventional shale reservoirs， were systematically summarized according to the characteristics and evaluation difficulties of reservoirs. For conventional sandstone reservoirs， a highly applicable identification technology for oil layers was systematically developed， and an innovative method for establishing electrical standards constrained by the lower limits of lithology and oil potential was developed under multi-layer joint testing conditions. In addition， a pioneering logging modeling technology was proposed based on different diagenetic zones. Various technologies were developed for complex reservoirs， including logging identification of sandstone lithology in steep slope zones， effectiveness evaluation of low-permeability sandstone reservoirs， and identification and evaluation of oil layers with low resistivity. For unconventional shale reservoirs， a logging evaluation technology of reservoir parameters was preliminarily developed to evaluate the storage properties and oil potential mainly based on two-dimensional nuclear magnetic resonance experiments. These logging evaluation technologies for different types of reservoirs reflected the combination of wellbore data and geological understanding， as well as the integration of rock physics and logging models， and they demonstrated the ability of logging to serve geology and reservoirs and achieved good application results. In the future， the remaining resources will mainly be distributed in hidden oil and gas reservoirs and deep layers. In order to meet the needs of exploration and development， it is necessary to strengthen the analysis of experimental data， gas logging， and testing production data， combine logging evaluation and geology， intensify the application of new logging technologies such as logging while drilling， imaging， and nuclear magnetic resonance， carry out comprehensive research and evaluation， and expand the evaluation and prediction of production capacity of oil layers from the perspective of evaluation objectives. Moreover， it is necessary to fully rely on expert experience to select samples and deepen the application of big data/artificial intelligence on the basis of conventional technologies， which will definitely achieve good results.

[Objective] Indium resources are an important supporting raw material for the development of China’s strategic emerging industries, and the study of the current status and dynamic evolution of the value flow of indium resources across the whole industry chain is of great strategic significance for the security of China’s indium resources and the sustainable development of its economy. [Methods] Based on the material flow framework of China’s indium resources in 2019, this paper utilizes the value chain analysis method and maps the value flow of China’s indium resources corresponding to the material flow process of indium resources according to the factor price theory, in order to evaluate and analyze the changes of value increase and decrease in the whole industry chain of China’s indium resources. [Results] (1) The total value of China’s indium resources industry chain shows an inverted U-shaped trend of increasing and then decreasing; (2) The stages with the highest value appreciation in the indium value chain are smelting and refining, with value increases of 922% and 1002% respectively. This is attributed to the substantial enhancement of indium resource grade due to the inputs of technology, labor, and energy, and that indium is a by-product metal with its associated zinc metal having relatively low value; (3) Although China is the world’s largest country in terms of indium resources, the indium resource industry chain still relies on imports to sustain its development; (4) Recycling of indium resources can have significant economic value. [Conclusion] Therefore, optimizing the indium resource industry chain, upgrading the production process and technology of high-end indium-containing products, and strengthening the recycling of indium-containing waste products are important ways to increase the value of indium resources in China.

The technology of power to hydrogen (P2H) and natural gas mixed hydrogen is highly regarded in theoretical research and engineering application in promoting the consumption of renewable energy and reducing carbon emissions. Targeting the park integrated energy system with high proportion renewable energy, an optimal configuration method of P2H considering natural gas mixed with hydrogen and cross seasonal storage is proposed. Firstly, the operational framework and energy flow relationship of the park integrated energy system are sorted out, and the mathematical models for the internal energy production, conversion, and storage equipment in the park are established. Secondly, a configuration model of P2H is established with the goal of optimizing the annual investment cost of equipment, the annual operating cost, and the carbon trading cost. Finally, the effectiveness of the proposed model is verified by an example analysis. The impact of changes in investment cost of electrolytic cells, the upper limit of mixed hydrogen volume fraction and cost weight coefficients of economy and low-carbon on the planned operation results are analyzed. The simulation results show that the proposed model can effectively improve the absorption capacity of renewable energy and reduce the overall economic cost and carbon emissions.

As an industrial chemical， bisphenol A is widely used in various food packaging materials. However， it is an endocrine disrupting chemical， which has estrogen activity and can cause certain damage to humans. So far， there are few studies on the detection of bisphenol A in self-heating food packaging materials， and there remains a lack of relevant standard. Therefore， it is necessary to establish a simple， sensitive and efficient method for the detection of bisphenol A in self-heating food. This study briefly introduces the pretreatment methods of bisphenol A， such as ultrasonic extraction， solid phase extraction， accelerated solvent extraction， and detection methods， such as gas chromatography-mass spectrometry， high performance liquid chromatography， fluorescent detection， and electrochemical detection.

Diopside[CaMg(SiO3)2] is a common mineral form of calcium magnesium silicate. Apart from dioside quarries, diopside also appears in skarn tailings. Diopside is a novel energy-saving raw material, mainly used in the ceramics industry. Glazed tiles prepared with diopside have the characteristics of low-temperature fast curing, which offers significant advantages to the building materials industry. The results reported in this paper show that the silicate and quartz composition in skarn lead and zinc tailings are likely to participate in the generation of ettringite and C-S-H gel in hydration reactions, respectively. Therefore, lead/zinc tailings can be used as concrete admixtures. As an important component of skarn tailings, the study of the ash reactivity of this type of tailings has great significance for comprehensive utilization in industry, but the relevant literature is incomplete. Paste samples were prepared with diopside, gypsum, and calcium hydroxide in this paper. The ash reactivity of fine-ground diopside was studied and hydration products were investigated using X-ray diffraction, scanning electron microscopy, fourier transform infrared spectroscopy, differential scanning calorimeter and nuclear magnetic resonance. The results show that the compressive strength of the paste prepared from fine-ground diopside can reach 9.83, 12.79, and 18.87 MPa at curing ages of 3, 7, and 28 d, suggesting that fine-ground diopside has good ash reactivity. The hydration products of cement prepared with fine-ground diopside are mainly accounted for by the C-S-H gel. Nuclear magnetic resonance results show that with the deepening of the hydration reaction, the percentage of silicon atoms in the Q2 structure state reduces and the Al/Si ratio in the C-S-H gel is lower than that in the original diopside material. With an increasing curing age, a small amount of gypsum and a large amount of Ca(OH)2 participate in the reaction. The amount of C-S-H gel hydration products increases gradually. The filling effect of the gypsum particles promotes the growth of the tructure's strength with an increasing curing time, although this effect does not alter the chemical reactions. These results will provide sufficient evidence for preliminary judgments of whether skarn tailings possess ash reactivity.

It is necessary to set up a purification stage in the middle of cascade for removing light impurities, which are formed in the process of isotope separation. These light impurities are harmful to the stable running of cascade if the impurity content is high and the cascade is long. The separation equations of a cascade were solved by a nonstationary method to investigate the feasibility of setting up a purge stage in the middle of the cascade. The effects of the purge stage, and the influence of the cut of the cascade and the purification ratio on the abundance of the light impurity and the target component were studied. The results indicated that the enrichment of the target component and the removal of the impurities could be accomplished simultaneously by a suitable choice of the cascade parameters when the purge stage was located in the middle of the cascade.

The paper analyzed main causes of longitudinal tearing accident of belt conveyor of coal mine, namely belt could be teared by anchor pole, puncheon, waste rock or roller, and proposed some preventing methods for longitudinal tearing of belt conveyor including strengthening routing inspection, using belt with pre-tearing net as far as possible, and setting protecting sensors of pre longitudinal tearing. The paper can provide certain guidance to prevent longitudinal tearing accident of belt conveyor of coal mine.

Objective: To review research progress on femoral attachment positioning during medial patellofemoral ligament (MPFL) reconstruction, so as to provide a reference for accurate positioning in clinic., Methods: The literature at home and abroad on femoral attachment positioning during MPFL reconstruction was extensively reviewed and summarized., Results: MPFL is the main ligament that restricts patellar outward migration, so MPFL reconstruction is the main treatment for patellar dislocation, but the accuracy of intraoperative femoral attachment positioning will significantly affect the effectiveness. At present, there are three main methods for femoral attachment positioning in MPFL reconstruction, including imaging positioning, bony landmark positioning, and new technology. Among them, the main imaging positioning method is the "Schöttle point" method, but it has high requirements for fluoroscopic positioning, and can only be accurately positioned under standard lateral fluoroscopy of the femur. The bony landmark positioning method mainly locates the femoral attachment by touching or dissecting the bony landmarks such as adductor tubercles and medial epicondyle of femur, but its disadvantages are that the positioning is not accurate enough, the intraoperative visual field exposure requirements are high, and a large incision is required. In order to avoid the problem that the simple bony landmark positioning method, in recent years, the combination of bony landmarks combined with arthroscopy, three-dimensional (3D) printing technology, and robot-assisted positioning methods have begun to be used in clinical practice. New technology localization methods have shown good results by preparing guides before operation, planning positioning paths in advance, or directly using robots to assist positioning during operation., Conclusion: The accurate positioning of the femoral attachment in MPFL reconstruction is crucial, and the method of accurate and rapid intraoperative determination needs to be further improved and optimized. In the future, it is expected that the combination of computer image recognition correction technology and intraoperative position assistance will solve this problem.

Objective: To investigate the mechanism of RNF7 for regulating myeloid-derived suppressor cells (MDSCs) in nonsmall cell lung cancer (NSCLC)., Methods: TIMER2.0 database and immunohistochemistry were used to analyze RNF7 expression level and its correlation with immune cell infiltration in non-small cell lung cancer. The impact of RNF7 expression levels on prognosis of lung cancer patients was analyzed using Kaplan-Meier survival analysis. CMT-167 cells with RNF7 overexpression or knockdown were inoculated subcutaneously in C57BL/6 mice, and the mice in RNF7 knockdown group were treated with anti-PD-1 or IgG isotype control 7 days after the inoculation. The tumor tissues were harvested after 30 days for tumor volume measurement, detection of S100A8+A9 and Gr-1 expressions with immunohistochemistry, and analysis of MDSC infiltration. Gene set enrichment analysis (GSEA) was performed to identify the potential pathways regulated by RNF7 in NSCLC. Western blotting and luciferase assays were used to assess the impact of RNF7 on the NF-κB signaling pathway. ELISA and RT-qPCR were used to measure chemokine (C-X-C motif) ligand 1 (CXCL1) expression., Results: RNF7 expression was significantly upregulated in NSCLC, and high RNF7 expression levels were associated with poor prognosis of the patients ( P < 0.001). TIMER2.0 analysis revealed a positive correlation between RNF7 expression and MDSC infiltration ( P < 0.001). GSEA suggested that RNF7 was enriched in the NF-κB signaling pathway. In NSCLC cells, RNF7 knockdown significantly inhibited NF-κB activation and reduced CXCL1 expression. In the tumor-bearing mice, RNF7 overexpression significantly increased MDSC infiltration in the tumor tissue, and RNF7 knockdown obviously reduced MDSC infiltration and enhanced the efficacy of anti-PD-1 therapy., Conclusion: High expression of RNF7 in NSCLC cells promotes CXCL1 expression by activating the NF-κB signaling pathway, thus leading to the chemotactic recruitment of MDSCs, which contributes to tumor resistance to antiPD-1 therapy.

As the special subject of the applicant for registration of medical device, the research and development institutions have insufficient conditions and abilities to become medical device registrants, and there are certain difficulties in the actual registration application process, such as not clearing the certification path for the research and development institutions to hold the certificate. In view of the existing problems, by comparing the path of medicine research and development institutions to become medical device registrants and combining with the actual medical device industry to give relevant suggestions, including improving quality management over the whole life cycle of medical devices, quality and safety responsibility ability of research and development institutions, establishing the registration and certification path of research and development institutions, supporting laws and regulations, etc., so as to ensure that the research and development institutions become medical device registrants successfully.

Objective: To investigate the effects of expression levels of S100 calcium-binding protein A10 (S100A10) in lung adenocarcinoma (LUAD) on patient prognosis and the regulatory role of S100A10 in lung cancer cell proliferation and metastasis., Methods: Immunohistochemistry was used to detect the expression levels of S100A10 in LUAD and adjacent tissues, and the relationship between S100A10 expression and clinicopathological parameters and prognosis of the patients was statistically analyzed. The lung adenocarcinoma expression dataset in TCGA database was analyzed using gene enrichment analysis (GSEA) to predict the possible regulatory pathways of S100A10 in the development of lung adenocarcinoma. Lactate production and glucose consumption of lung cancer cells with S100A10 knockdown or overexpression were analyzed to assess the level of glycolysis. Western blotting, CCK-8 assay, EdU-594 assay, and Transwell assays were performed to determine the expression level of S100A10 protein, proliferation and invasion ability of lung cancer cells. A549 cells with S100A10 knockdown and H1299 cells with S100A10 overexpression were injected subcutaneously in nude mice, and tumor growth was observed., Results: The expression level of S100A10 was significantly upregulated in LUAD tissues as compared with the adjacent tissues, and an elevated S100A10 expression level was associated with lymph node metastasis, advanced tumor stage and distant organ metastasis ( P < 0.05), but not with tumor differentiation or the patients' age or gender ( P > 0.05). Survival analysis showed that elevated S100A10 expressions in the tumor tissue was associated with a poor outcome of the patients ( P < 0.001). In the lung cancer cells, S100A10 overexpression significantly promoted cell proliferation and invasion in vitro ( P < 0.001). GSEA showed that the gene sets of glucose metabolism, glycolysis and mTOR signaling pathway were significantly enriched in high expressions of S100A10. In the tumor-bearing nude mice, S100A10 overexpression significantly promoted tumor growth, while S100A10 knockdown obviously suppressed tumor cell proliferation ( P < 0.001)., Conclusion: S100A10 overexpression promotes glycolysis by activating the Akt-mTOR signaling pathway to promote proliferation and invasion of lung adenocarcinoma cells.

Cerium-manganese modified biochar (MBC) combined with earthworms ( Eisenia foetida) can immobilize arsenic (As) in red soils. In this study, high-throughput sequencing technology was used to explore the combined effects of MBC and E. foetida on bacterial diversity and community structure in As-contaminated red soils. The results showed that the single earthworm treatment had the highest diversity index, whereas the diversity index decreased in the single biochar or MBC treatment, indicating that earthworms can boost the growth of bacteria in the soil, and the addition of biochar/MBC all decreased the bacterial diversity of soils. When biochar/MBC was combined with earthworms, the diversity index increased to some degree. In terms of bacterial community structure, the relative abundance of Proteobacteria and Bacteroidetes increased significantly in each treatment, especially for MBC-earthworm treated soil, in which the relative abundance was increased by 17.08% and 329.47% for Proteobacteria and Bacteroidetes, respectively, compared to that in the control (CK). Otherwise, those abundances were decreased by 19.18% and 48.76%, respectively, for Acidobacteria and Chloroflexi. Correlation analysis results showed that the soil water-soluble As (WSAs) was negatively correlated with the relative abundances of Proteobacteria and Bacteroides ( P <0.05) but was positively correlated with Acidobacteria and Chloroflexi ( P <0.05), which indicated that with the decrease in WSAs in soils, the bacteria of Proteobacteria and Bacteroides reproduced rapidly, whereas the Acidobacteria and Chloroflexi were inhibited. Moreover, different treatments induced selective changes in the bacterial community, in which earthworms significantly promoted the proliferation of γ -Proteobacteria, Flavobacteriales, Aeromonadales, and Variovorax and earthworms improved the immobilization effect of As by promoting the growth of these bacteria.

The development and implement of microbial chassis cells can provide excellent cell factories for diverse industrial applications, which help achieve the goal of environmental protection and sustainable bioeconomy. The synthetic biology strategy of Design-Build-Test-Learn (DBTL) plays a crucial role on rational and/or semi-rational construction or modification of chassis cells to achieve the goals of "Building to Understand" and "Building for Applications". In this review, we briefly comment on the technical development of the DBTL cycle and the research progress of a few model microorganisms. We mainly focuse on non-model bacterial cell factories with potential industrial applications, which possess unique physiological and biochemical characteristics, capabilities of utilizing one-carbon compounds or of producing platform compounds efficiently. We also propose strategies for the efficient and effective construction and application of synthetic microbial cell factories securely in the synthetic biology era, which are to discover and integrate the advantages of model and non-model industrial microorganisms, to develop and deploy intelligent automated equipment for cost-effective high-throughput screening and characterization of chassis cells as well as big-data platforms for storing, retrieving, analyzing, simulating, integrating, and visualizing omics datasets at both molecular and phenotypic levels, so that we can build both high-quality digital cell models and optimized chassis cells to guide the rational design and construction of microbial cell factories for diverse industrial applications.

Objective: The estrogen level and blood calcium concentration changes were studied on menopausal women with benign paroxysmal positional vertigo (BPPV). Methods: Between January 2015 and January 2016, 70 menopause women with BPPV in outpatient clinics of Department of Otorhinolaryngology, Inner Mongolia Medical University Affiliated Hospital were included in this study as research group, while 30 menopause healthy women who came to hospital for check-up were included as control group. Serum levels of estradiol (E2), follicle stimulating hormone (FSH), luteinizing hormone (LH), progesterone (PRO), testosterone (T), serum prolactin (PRL) and the calcium concentration were analysed and comparied between research group and control group. SPSS17.0 statistical software was used to analyze the data. χ(2) test was used to compare the percentage of decreased serum level of sex hormone, and t test was used to compare the serum level of sex hormone and calcium concentration of two groups. Results: In research group, sex hormone decreased proportion of E2 (91%) and PRO (67%) were obviously higher than those in control group (χ(2) value was 8.13, 10.28, respectively, all P <0.05). The E2 and PRO in research group were significantly lower than those in control group ((33.18±31.45) pmol/L vs (64.92 ±31.52) pmol/L, (0.64±0.48) nmol/L vs (1.02±0.60) nmol/L, t value was 6.238, 8.566, respectively, all P <0.05). There was no statistically significant difference of the level of LH, PRL, T, FSH and blood calcium concentration in research group compared with control group ((29.81±13.13) U/L vs (27.21±10.19) U/L, (0.49±0.20) nmol/L vs (0.49±0.15) nmol/L, (0.56±0.42) nmol/L vs (0.73±0.62) nmol/L, (64.25±31.44) U/L vs (60.38±29.97) U/L, (2.28±0.17) mmol/L vs (2.32±0.21) mmol/L, t value was 13.427, 14.876, 7.505, 12.090, 7.532, respectively, all P >0.05). Conclusion: The level of E2 and PRO decrease obviously in postmenopausal women with BPPV, which can cause the inner ear microcirculation disorder , may be one of the risk factors of BPPV.

Hydrogen sulfide (H2S)is a novel gastrotransmitter, and plays a protective role in many diseases. Adipocyte endogenously produces H2S by cystathionine β synthase (CBS), cystathionine γ lyase (CSE) and 3-mercaptopyruvate sulfurtransferase (MPST).The endogenous H2S in adipocyte plays an essential role in glucose uptake activity and utilization, lipolysis and adipocyte differentiation, and then takes part in the pathogenesis of obesity, diabetes and cardiovascular diseases.H2S regulates adipocyte energy metabolism by activation activating insulin receptor, peroxisome proliferator activated receptor γ (PPARγ) or potassium channel. H2S acts as an "energy switch" in regulation of metabolic diseases.

Cardiovascular disease is a common disease that threatens the health of human beings,espe-cially elders.The number of people dying from cardiovascular diseases annually ranks top one in the world-wide.Long noncoding RNAs (LncRNAs)is a big group of ncRNAs with nucleotides length >200 nt,and its numbers is most in the body.More and more study found that LcRNAs play critical roles in the pathogenesis and development of cardiovascular disease.This review will describe the latest research progress of long noncoding RNA in cardiovascular diseases.

Objective: To study the relationship between knee osteoarthritis (OA) and intercondylar notch narrowing based on the notch width index., Methods: Magnetic resonance (MR) images were collected from middle-aged and elderly patients with a definite diagnosis of knee OA, including 42 with mild OA and 37 with moderate to severe OA, with 70 healthy individuals serving as the control group. The notch width indexes NWI, NWI-A, and NWI-P on the coronal images at different levels were calculated, and the intercondylar notch was classified, according to the features on axial MR images, into types A, U, and W. The association of OA with NWI, NWI-A, NWI-P, and notch type was determined, and the cutoff values were obtained based on the ROC curves at different levels as indicators for diagnosis of intercondylar notch stenosis., Results: In the control, mild OA, moderate to severe OA groups, the NWI value on coronal MR images were 0.252±0.019, 0.251±0.017, and 0.240±0.020, NWI-A were 0.261±0.024, 0.259±0.023, and 0.245±0.023, and NWI-P were 0.271±0.026, 0.270±0.024, and 0.254±0.022, respectively. Patients with moderate to severe OA had significantly smaller NWI, NWI-A, and NWI-P than the other two groups (P<0.05), and a significant association was found between NWI values at each level and the occurrence of moderate to severe OA (P<0.01). A NWI value<0.248, NWI-A<0.256, and NWI-P<0.266 supported a diagnosis of intercondylar notch narrowing. Type A intercondylar notch was found in the majority of patients with intercondylar notch narrowing (P<0.05)., Conclusion: Patients with moderate to severe OA have significant intercondylar notch narrowing, and patients with a type A intercondylar notch are more likely to have intercondylar notch narrowing than those with type U notch.

Objective: To observe the vasodilating effect of adrenomedullin 2 (ADM2) by antagonizing angiotensin 1 (Ang II), and to explore its mechanism., Methods: Eighteen male, 180-200 g SD rats were randomly divided into 3 groups (n = 6): control group, Ang II (150 ng/(kg x min)) group and Ang II (150 ng/(kg x min)) + ADM2(500 ng/(kg x h)) group. Mini-osmotic pumps filled with peptide were implanted in the back of rats subcutaneously. After two weeks, the blood pressure was measured by the way of carotid intubation. The plasma was collected for the detection of nitric oxide (NO) content and the activity of endothelial nitric oxide synthase (eNOS). The in situ oxidation of fluorescent dye dihydroethidium (DHE) was used for detecting superoxide in rat arteries. The rat isolated arterial rings were made for studying the vasodilating effect of ADM2. Human umbilical vein endothelial cell line EA. hy 926 cells were cultured and their intracellular reactive oxygen species (ROS) were evaluated by probe DCFH-DA., Results: ADM2 dramatically decreased the blood pressure in angiotensin II-induced hypertension rat model, enhanced plasma NO content and the activity of eNOS and reduced superoxide formation in vessel walls. ADM2 also induced relaxation of the vascular rings preconstricted by Ang II in a concentration-dependent and endothelium-dependent manner. In cultured vascular endothelium, ADM2 ameliorated the ROS generation induced by Ang II., Conclusion: Adrenomedullin 2 relaxed blood vessels by antagonizing angiotensin II-induced oxidative stress and improving the vascular endothelial function.

Objective: To evaluate the clinical value and specificity of atrial natriuretic peptide (ANP) in early diagnosis of sepsis., Methods: A prospective study was performed. Data of patients with sepsis were consecutively collected from September 2007 to December 2012 according to the international criteria for the diagnosis of sepsis, and that of 114 patients admitted to intensive care unit (ICU) and the cadre health care ward were divided into three groups: systemic inflammatory response syndrome (SIRS) group (n=37), sepsis group (n=41) and severe sepsis group (including severe sepsis and septic shock, n=36). Venous blood of each patient was drawn instantly when admitted to ICU. The concentration of plasma ANP in each group was determined on the 1st day using a new type of sandwich immunofluorescence assay, and other biomarkers, such as procalcitonin (PCT, detected with double antibody immunochemi luminometry), blood lactic acid (detected with electrode meter), and C-reactive protein (CRP, detected with immunonephelometric analysis), and the acute physiology and chronic health evaluation II (APACHEII) score was recorded. APACHEII score and biomarkers were compared among three groups. The risk factor for severity was confirmed with stepwise regression, and the value of each index in early diagnosis of sepsis was analyzed with receiver operating characteristic (ROC) curve., Results: The plasma concentration of ANP exhibited a gradual increase with the aggravation of the disease, and the median ANP value was found to be highest in the severe sepsis group compared with SIRS group and sepsis group [μg/L: 0.26 (0.22) vs. 0.19 (0.05), 0.21 (0.08), P<0.01 and P<0.05]. The regression equation was established at y=0.69 + 0.66APACHEIIscore + 1.285 ANP value, with group as independent variable, and APACHEII score, ANP, PCT, CRP and blood lactic acid value as dependent variables. It could be concluded that APACHEII score and ANP value were correlated with severity of sepsis, and accordingly they were regarded as the independent predictors of severity of sepsis. It was found through the ROC curve analysis of ANP in the sepsis patients, that the area under the ROC curve for ANP [0.805, P=0.000, 95% confidence interval (95%CI) 0.726-0.883, sensitivity 75.8%, specificity 78.4%] was similar with the area under the APACHEII score (0.820, P=0.000, 95%CI 0.742-0.897, sensitivity 68.4%, specificity 78.4%), and it was apparently higher than AUCs of PCT (0.716, P=0.000, 95%CI 0.622-0.810, sensitivity 67.1%, specificity 62.2%), CRP (0.569, P=0.236, 95%CI 0.463-0.675, sensitivity 76.3%, specificity 41.5%), or blood lactic acid (0.566, P=0.254, 95%CI 0.453-0.679, sensitivity 75.0%, specificity 48.6%)., Conclusions: Plasma ANP concentration is clinically valuable in early diagnosis and severity assessment of sepsis, and it is more specific and sensitive than biomarkers PCT and CRP.

Objective: To establish a simple method of measuring hydrogen sulfide (H2S) in cultured living cells., Methods: Filtration membrane was stuck on the lid of cell culture plate. H2S released from cultured cells was trapped by zinc acetate to generate ZnS deposition. Then the ZnS trapped in the filtration membrane was measured by methylene blue assay and the H2S production from the living cells was counted according to the standard curve. This simple method was used to access the H2S release in HepG2 (high expression CBS and CSE) and HUVEC (low expression CSE) cell lines., Results: H2S generation in cultured HepG2 cells assayed using the present method was (859.39±19.12) nmol/(min×10(6) cells). PAG (CSE inhibitor), HA (CBS inhibitor) or the two-inhibitor (PAG+HA) treatment significantly lowered H2S release, respectively: (341.34±105.90) nmol/(min×10(6) cells), (375.05±174.50) nmol/(min×10(6) cells), and (204.47±97.14) nmol/(min×10(6) cells). The H2S production of HUVEC was (26.23±3.24) nmol/(min×10(6) cells) (about 1/30 production of HepG2 cell). Trypan blue assay showed that the cell viability was greater than 95%, suggesting that there was no cytotoxicity by using the present instrument., Conclusion: The modified instrument in cell culture plate lid was feasible for detection of hydrogen sulfide release in living cells.

Objective: To investigate the role of hydrogen sulfide (H₂S) on oxidized-low density lipoprotein (ox-LDL)-stimulated NF-κB pathway in human monocytes/macrophages and its mechanisms., Methods: THP-1 cells were induced to differentiate into macrophages by incubation with phorbol myristate acetate (PMA) . The human monocytes/macrophages were divided into 4 groups: control group, ox-LDL group, ox-LDL + H₂S 100 μmol/L group and ox-LDL + H₂S 500 μmol/L group. The expression of IκBα and phosphorylation of NF-κB p65 in the cells were detected by Western blotting. The expression of IκBα and nuclear translocation of NF-κB p65 in the cells were observed by laser confocal method. The interaction between NF-κB p65 and IκBα in the nuclear extracts was detected by coimmunoprecipitation method., Results: Compared with the control group, the phosphorylation of NF-κB p65 in the human monocytes/macrophages of ox-LDL group was increased significantly (0.855 ± 0.116 vs. 0.502 ± 0.218, P=0.046), while the expression of IκBα in the cells of the ox-LDL group was decreased (0.612 ± 0.216 vs. 0.997 ± 0.167, P=0.029). Compared with the ox-LDL group, the phosphorylation of NF-κB p65 in the cells of the ox-LDL + H₂S 100 μmol/L group and the ox-LDL + H₂S 500 μmol/L group was decreased significantly (0.424 ± 0.225 vs. 0.855 ± 0.116, P=0.020; 0.378 ± 0.071 vs. 0.855 ± 0.116, P=0.011, respectively), while the expressions of IκBα in the cells of the ox-LDL + H₂S 100 μmol/L group and the ox-LDL + H₂S 500 μmol/L group were increased (1.037 ± 0.111 vs. 0.612 ± 0.216, P=0.015; 1.046 ± 0.084 vs. 0.612 ± 0.216, P=0.013, respectively). The results from laser confocal method demonstrated that the IκBα expression in the cytoplasma of cells in the ox-LDL group was lower than that in the control group, and the nuclear translocation of NF-κB p65 in the cells of the ox-LDL group was higher than that in the control group. The IκBα expression in the cytoplasma of cells in the ox-LDL+ H₂S 100 μmol/L group and ox-LDL + H₂S 500 μmol/L group was higher than that in the ox-LDL group, and the nuclear translocation of NF-κB p65 in the cells of ox-LDL group was lower than that in the ox-LDL group. The coimmunoprecipitation experiment showed that no IκBα integrated with NF-κB p65 was detected in the nuclear extracts of cells in the control group, a small amount of IκBα integrated with NF-κB p65 was detected in the nuclear extracts of cells in the ox-LDL group, but a large amount of IκBα integrated with NF-κB p65 was detected in the nuclear extracts of cells in the ox-LDL+H₂S 100 μmol/L group and the ox-LDL + H₂S 500 μmol/L group., Conclusion: H₂S inhibited the activation of NF-κB p65 pathway in the ox-LDL-induced human monocytes/macrophages. The mechanisms might involve the prevention of the degradation of IκBα, then the inhibition of the phosphorylation and nuclear translocation of NF-κB p65, thus promoting the IκBα integrated with NF-κB p65 in the nuclei, and then inhibiting the activity of NF-κB.

In past decade, hydrogen sulfide (H₂S) as a novel gasotransmitter, covered many fields in biological and medical research. However, there is no effective, convenient and high-throughput method for determination of circulatory H₂S until now. Here, we aim to develop an easy method for measurement of circulatory H₂S by modified methylene blue method. In the present study, we added Zn²⁺ to plasma sample to deposit H₂S, HS⁻ and S²⁻, as well as plasma protein, then used NaOH to re-dissolve plasma protein. ZnS deposition was re-dissolved by the addition of N, N-dimethyl-p-phenylenediamine, and the remnant protein was deposited by trichloroacetic acid. After centrifugation, ferriammonium sulfate was added to the supernatant fluid to generate methylene blue, which was analyzed by spectrophotometer at 665 nm. Using the present method, we found that most ions including sulfate and thiosulfate did not affect detection of H₂S concentration, but albumin (physiological concentration) reduced the detection value, which suggested the binding of serum albumin and a certain amount of H₂S. The relative recovery ratio of present method is 81.9%, which implies that the method is relative accurate for the determination of H₂S concentration in plasma or serum. H₂S levels of frozen plasma samples from 65 healthy volunteers detected by the present method were (13.93 ± 4.98) µmol/L. There was no obvious difference between the detection values of fresh and frozen samples from the same SD rats. These results suggest the modified methylene blue assay is stable, sensitive, convenient and high-throughput. The method can be used to analyze the circulatory H₂S in clinical and basic research.

In order to ensure high stability and strong anti-interference ability in static interference system for qualitative and quantitative analysis of gas, a static scans interference detection system was designed based on photoelastic modulation infrared spectrum absorption system. The system consists of infrared laser, polarizer, photoelastic modulator, polarization analyzer and CCD components. By photoelastic modulator the principal refractive index of optical crystal will change cyclically by the modulation signal, producing cyclical changes in the optical path difference. With the calculation of modulation phase variation, the authors can get the function of the crystal length, the modulation cycle, and the range of optical path difference. Based on phase delay value and the energy distribution of interference pattern, the authors got the formula for the corresponding interference light intensity. The experiment used ZnSe crystal as the photoelastic modulation crystal, the polarizer uses the DOP3212 polarizer, and the detector uses the TCD5390AP array CCD. The five groups have different concentrations with three common VOC gases (formaldehyde, benzene and xylene) for detecting the concentrations of gases. The experimental results with the traditional infrared absorption were compared with the test results of photoelastic modulation infrared spectrum absorption method. The method of photoelastic modulation infrared spectrum absorption had high stability and real-time features, while the detection accuracy is better than the traditional infrared absorption method.

Objective: To improve a method of a new hydrogen sulfide slow-releasing donor, to observe its cellular toxicity in HepG2 cells and tissue distribution and metabolic pathway after administration of the donor by intraperitoneal injection in ICR mice and to afford experimental evidences for rationally using this donor in hydrogen sulfide research., Methods: We synthesized the new chemical compound which slowly released hydrogen sulfide. After administration of the donor, the cell toxicity was evaluated for cell viability using trypan blue staining in HepG2 cells and lactate dehydrogenase (LDH) activity in culture medium. After administration of this donor by intrasperitoneal injection, we measured the tissue hydrogen sulfide content in the liver, heart, kidney and brain using sensitive-sulfur electrode assay., Results: We successfully prepared the donor which could release hydrogen sulfide. The releasing ability of the donor solution stored at 4 °C or 20 °C did not change as compared with the freshly-prepared one. Treated at various concentrations of the donor (0.062 5, 0.125, 0.25, 0.5, 1 and 2 mmol/L) for 24 hours in HepG2 cells, the cell viability and LDH leak from the cells were not different as compared with the controls. The donor (2 mmol/L) was administrated everyday and the culture medium was changed every 3 days. After 9 days, the cell viability and LDH leak did not change. Administration of the donor (200 μmol/kg) quickly increased the tissue hydrogen sulfide concentrations in the liver and heart and maintained about 20 min; the hydrogen sulfide level in the kidney elevated and maintained a longer time, then recovered after 2 hours, which implicated that the donor might exclude the kidney; the hydrogen sulfide concentration in the brain did not change in the present study, which suggested that the donor could not pass the blood-brain barrier. Long time (4 weeks) treatment with this compound might induce hepatic or cutaneous injury., Conclusion: The new chemical compound is a relative stable, slow-releasing donor of hydrogen sulfide with low cellular toxicity, which may be used to study the regulatory role of hydrogen sulfide in the cellular physiological and pathophysiological mechanism of the animal model with acute diseases.

Objective: Adenosine triphosphate (ATP) can promote the repair of spinal cord injury (SCI). To investigate the effect of ATP combined with bone marrow mesenchymal stem cells (BMSCs) transplantation on SCI, and to evaluate the synergistic action of ATP and BMSCs in the repair of SCI and the feasibility of the combined transplantation in the treatment of SCI., Methods: BMSCs were isolated from the marrow of the tibia and the femur of a male SD rat (weighing 120 g), the 3rd generation BMSCs were labeled with BrdU, then BMSCs suspension of 5.0 x 10(7) cell/mL were prepared. Forty-eight adult female SD rats (weighing 240-260 g) were made SCI models at T2 levels according to the improved Allen's method, and were randomly divided into 4 groups (groups A, B, C, and D, n = 12). In group A, ATP (40 mg/kg) and BMSCs (6 microL) were injected to the central point and the other 2 points which were 1 mm from the each side of head and tail of the injured spinal cord; after blending the BMSCs suspension, the cells amount was about 3.0 x 10(5). In groups B, C, and D, the BMSCs suspension (6 microL), ATP (40 mg/kg), and PBS (40 mg/kg) were injected to the points by the same method as group A, respectively. The general conditions of the rats were observed after operation. The nerve function of low extremities was evaluated using the improved Tarlov scale and the Rivlin inclined plane test at 1, 3, 7, 14, 21, and 28 days after operation. At 28 days after operation, the reparative effect of SCI was observed using histological and immunohistochemical staining., Results: One rat of group A, 2 of group B, 2 of group C, and 3 of group D died of infection and anorexic, the others survived to the end of the experiment. Paralysis symptom in low extremities occurred in all rats after operation and was improved at 2-3 weeks postoperatively, the improvement of group A was the best, groups B and C were better, group D was the worst. There was no significant difference in the Tarlov scale and the Rivlin inclined plane test among 4 groups at 1 and 3 days after operation and between groups B and C at 7, 14, 21, and 28 days after operation (P > 0.05), but there were significant differences among other groups at 7, 14, 21, and 28 days after operation (P < 0.05). At 28 days after operation, HE staining demonstrated that the injured region in group A was finely restored, without obvious scar tissue and cavity, and there existed clear stem cell differentiation characters; there was small amount of scar tissue and cavity in the injury site of groups B and C; and there was great deal of scar tissue in the injury site of group D, in which there were numerous inflammatory cells and fibroblasts infiltration and bigger cavity. Immunohistochemical staining showed that BrdU-positive BMSCs were seen in groups A and B, and positive cells of group A was significantly more than that of group B (P < 0.05). The expressions of neurofilament protein 200 and glial fibrillary acidic protein in group A were significantly higher than those in groups B, C, and D, and groups B and C were significantly higher than group D (P < 0.05)., Conclusion: ATP has protective effects on injured spinal cord, a combination of ATP and BMSCs can synergistically promote the reparation of SCI.

Objective: To explore atrioventricular connection and atrioventricular segmental situs in patients with crisscross heart (CCH) and to evaluate the diagnostic value of echocardiography for this anomaly., Methods: Ten consecutive patients with crisscross heart were enrolled into this retrospective study. Their echocardiographic data were analyzed and compared with the results of X-ray angiocardiography and 64-slice multi-detector row computed tomography (MDCT) or MRI., Results: The crossing of atrioventricular valves could be seen in each case by scanning in a subxiphoid or apical 4-chamber view. Both the positive rate and the specificity were 100%. Horizontal ventricular septum was in 9 cases and vertical (sagittal) ventricular septum in 1 case. The segmental set of 8 patients with concordant atrioventricular connection was {S. D. L} in 5 cases, {S. D. D} 1 case, {S. D. S} 1 case and {S. L. D} 1 case. The segmental set of 1 case with discordant atrioventricular connection was {I. D. D} and another 1 case with ambiguous atrioventricular connection was {A. L. L}. In 1 case, the atrioventricular connection was inconsistent with the atrioventricular segmental situs. Ventriculoarterial connections were concordant in 1, DORV in 6, TGA in 2 and C-TGA in 1., Conclusion: Echocardiography is proven quite helpful in diagnosis of CCH, and continuous sweeps in subxiphoid long-axis plane or apical 4-chamber view play a key role. Both the atrioventricular connection and the atrioventricular segmental situs are complicated so that they are not always concordance with each other. It is necessary to account for separately.

Objective: To investigate the role of endogenous hydrogen sulfide (H2S) in patients with asthma., Methods: Levels of serum H2S, lung function and cell differential count in induced sputum were studied in 44 patients with acute exacerbation of asthma, 33 patients with stable asthma and 12 health subjects. Of the 33 patients with stable asthma, 3 failed to achieve induced sputum., Results: The serum H2S level was (75.2+/-13.0) micromol/L in controls (12 cases), (55.8+/-13.6) micromol/L in patients with stable asthma (33 cases), (57.8+/-6.3) micromol/L in patients with mild of acute exacerbation asthma (9 cases), (40.8+/-5.1) micromol/L in that with moderate of acute exacerbation asthma (13 cases), and (31.3+/-2.9) micromol/L in that with severe of acute exacerbation asthma (22 cases, F = 44.592, P < 0.01). The level of serum H2S was significantly lower in smokers (45.4+/-19.5) micromol/L as compared with nonsmokers (52.7+/- 16.0) micromol/L (F = 4.804, P < 0.05). Status of the subjects (in good health, stable asthma, and exacerbation of asthma) was positively correlated with H2S in serum (r = 0.712, P < 0.01). H2S in serum was positively correlated with FEV1.0 % (r = 0.877, P < 0.01) and the percentage of sputum macrophages (r = 0.791, P < 0.01); and it was negatively correlated with count of sputum cells (r = -0.348, P < 0.01) and the percentage of sputumneutrophils (r = -0.906, P < 0. 01)., Conclusion: Endogenous H2S may be involved in the pathogenesis of airway obstruction in asthma and may be a noninvasive marker of disease activity and severity.

Objective: To observe the effects of sulfur dioxide (SO2) on cardiac function of isolated perfused heart of rat and to explore the physiological regulation of endogenous SO2 on myocardial action., Methods: The hearts of 64 Wistar rats were isolated, perfused with Krebs-Henseleit (KH) solution through Langendorff apparatus, and randomly divided into 8 equal groups: Four groups underwent perfusion of SO2 of the concentrations 1, 10, 100, 1000 micromol/L respectively for 5 min and then perfused with KH solution for 15 min. Eight hearts underwent perfusion of SO2 of the physiological concentration (10 micromol/L) for 20 min. The control group underwent perfusion of KH solution for 20 min. Eight hearts of the nicardipine group underwent perfusion of nicardipine, a L-type calcium channel blocker, 2.5 micromol/L for 5 min, SO2 10 micromol/L for 5 min, and then KH solution for 10 min. The heart in the hydroxamate (HDX) group underwent perfusion of HDX, an inhibitor of SO2 endogenous generating enzymes, for 5 min, and then perfused by KH solution for 15 min. The heart rate (HR), difference of left ventricular pressure (LVP), left ventricular peak rate of contraction (+ dp/dtmax), peak rate of relaxation (- dp/dtmax), and coronary flow (CF) were measured. Then transmission electron microscopy was conducted., Results: SO2 concentration-dependently inhibited the left ventricular +/- dp/dtmax, LVP, HR, and CF (all P < 0.01). The left ventricular +/- dp/dtmax, LVP, and HR were inhibited (P < 0.05) by the physiological concentration (10 micromol/L) SO2 donor continuous perfusion for 20 min. During perfusion 20 min, the LVP, + dp/dtmax, - dp/dtmax, and HR after perfusion for 20 min of the physiological concentration (10 micromol/L) SO2 donor continuous perfusion group were (15 +/- 3) mm Hg, (485 +/- 74) mm Hg/s, (339 +/- 64) mm Hg/s, and (114 +/- 26)/min respectively, all significantly lower than those 5 min after perfusion [(23 +/- 7) mm Hg, (595 +/- 93)mm Hg/s, (436 +/- 83) mm Hg/s, and (159 +/- 31)/min, all P < 0.05]. The LVP, + dp/ dtmax, -dp/dtmax, and HR of the nicardipine group were(37 +/- l0)mm Hg, (1025 +/- 287)mm Hg/s, (570 +/- 181)mm Hg/s, and (139 +/- 48)/min respectively, all not significantly different from those of the control group. The LVP, + dp/dtmax, - dp/dtmax, and CF after perfusion of the HDX group were (50 +/- 11)mm Hg, (1167 +/- 270) mm Hg/s, (889 +/- 72) mm Hg/s, and (6.3 +/- 1.9) ml/min respectively, all significantly lower than those before perfusion [(69 +/- 16) mm Hg, (1579 +/- 315) mm Hg/s, (1186 +/- 263) mm Hg/s, and (9.5 +/- 1.3) ml/min, all P < 0.05]., Conclusion: Exogenous SO2 has negative inotropic effect on myocardium. by the mechanism related to voltage-gated calcium channel. Nicardipine blocks the inhibitory effect of SO2 at physiological concentration.

Objective: To construct a detection method of endogenous hydrogen sulfide (H2S) from erythrocytes., Methods: Prepared rat erythrocyte (10(6) cell) was added in 4 mL 2-amino-2-methyl-1,3-propanediol(225 mmol/L, pH=9.55) and ultrasonically lysed for 3 times. The erythrocyte lysis was transferred into a 25 mL Erlenmeyer flask and beta-mercaptopyruvate was added for final concentration of 2 mmol/L. Cryovial test tubes (2 mL) were used as the centre wells each contained 0.5 mL of 1% zinc acetate as trapping solution and a filter paper of 2.0x2.5 cm2 to increase the air/liquid contacting surface. The flasks containing reaction mixture and centre wells were flushed with N2 before being sealed with a double layer of Parafilm. Reaction was initiated by transferring the flasks from ice to a 37 degrees C shaking water bath. After incubation at 37 degrees C for 60 min, 0.5 mL of 50% trichloroacetic acid was added into the reaction mixture to stop the reaction. The flasks were sealed again and incubated at 37 degrees C for another 60 min to ensure a complete trapping of the H2S released from the mixture. Released H2S is absorbed by zinc acetate and generation zinc sulfide. The zinc sulfide formed is dissolved in a hydrochloric acid solution of amino-dimethylaniline (N,N-dimethyl-p-phenylenediamine), and methylene blue is formed within 10 min at room temperature in the presence of ferric chloride. The blue color of methylene blue is measured at 670 or 650 nm on spectrophotometer., Results: We first demonstrated the key enzymes of endogenous H2S generation-3-mercaptopyruvate sulfurtransferase (MPST) gene expression by RT-PCR. Endogenous H2S production from rat erythrocytes was 22.76+/-1.53 micromol/min/10(8) cells, about 4-folds as compared with to the liver and kidney tissues. However, the endogenous H2S production from erythrocyte by L-cysteine pathway was little detected., Conclusion: Endogenous H2S released from erythrocytes depended mainly on MPST pathway. Our method is effective to detect the erythrocytic endogenous H2S.

Objective: To study the changes of endogenous hydrogen sulfide (H(2)S) and the effect of exogenously applied H(2)S on ovalbumin-induced acute asthma., Methods: Twenty-four Male SD rats were randomly divided into a control group (n = 8), an asthma group (n = 8) and a NaHS group (n = 8). Pulmonary function was measured, and the pulmonary pathology changes, the content of H(2)S in lung tissue and plasma and the activity of H(2)S generating enzymes in lung tissue were detected at the 28 th day after ovalbumin administration. Western blotting was used to detect the endothelial cystathionine-gamma-lyase CSE protein in the lung tissues., Results: In the asthma group, the peak expiratory flow (PEF) was (2.90 +/- 0.70) L/s, the contents of H(2)S in the plasma was (10 +/- 3) micromol/L, in the lung tissue was (4.9 +/- 1.3) micromol/L. The H(2)S generating enzyme activity in the lung tissue of the asthma group was (1.00 +/- 0.10) nmolxmin(-1)xmg(-1) pro, and the lung CSE content of the asthma group was significantly lower than that of the control group (6.50 +/- 0.10) L/s, (54 +/- 10), (24.1 +/- 8.0) micromol/L, (1.80 +/- 0.10) nmolxmin(-1)xmg(-1), F = 112.13, 110.10, 27.34, 79.39, 12.28, all P < 0.05). The pulmonary pathology score of the asthma group was 3 (2 - 4), significantly higher than that of the control group [1 (0 - 1), H = 16.93, P < 0.01]. In the NaHS group, the PEF was (5.70 +/- 0.50) L/s, the content of H(2)S in the plasma was (17 +/- 4) micromol/L, in the lung tissue was (15.3 +/- 4.0) micromol/L, the H(2)S generating enzyme activity in the lung tissue was (1.60 +/- 0.20) nmolxmin(-1)xmg(-1) pro, the lung CSE content of the NaHS group was significantly higher than that of the asthma group (F = 112.13, 110.10, 27.34, 79.39, 12.28, all P < 0.05). The pulmonary pathology score of the NaHS group was 1 (1 - 2), significantly lower than that of the asthma group [3 (2 - 4) score, H = 16.93, P < 0.01]. There was a significantly positive correlation between the content of the content of H(2)S in lung tissue and PEF (r = 0.74, P < 0.01). There was a significantly negative correlation between the content of H(2)S in lung tissue and the pulmonary pathology score (r = -0.64, P < 0.01)., Conclusion: Endogenous H(2)S is involved in the pathogenesis of asthma in this animal model. Exogenously applied H(2)S can attenuate inflammation of asthma and exert protective effect from asthma.

Thirty microsatellite markers were used to analyze the genetic diversity of five Silver carp populations in the middle and lower reaches of the Yangtze River. A total of 144 different alleles were found and the number of alleles in each locus ranged from 1 to 10. Twenty-five loci (83.33%) were polymorphic. In the five populations, the average number of alleles was 4.0 to 4.1, the number of mean valid alleles was 2.4445 to 2.6332, the value of average observed and expected heterozygosity ranged from 0.3233 to 0.3511 and 0.4421 to 0.4704, and the mean PIC was 0.4068 to 0.4286. Fst value indicated that the populations were moderately differentiated. Chi-square test was used to analyze the genotypes based on Hardy-Weinberg equilibrium, the P value denoted that the five populations deviated equilibrium partially. The genetic similarity coefficient of the five populations was 0.8466 to 0.9146, and the genetic distance of the populations was 0.0893 to 0.1665. A UPGMA dendrogram was constructed based on Nei's standard genetic distance for the five Silver carp populations.

Objective: To investigate alteration and cross link of the aortic intima and platelet endogenous L-Arg/NOS/NO pathway induced by water immersion restraint (WIR) stress., Methods: After 7 h of WIR stress, the aortic intima was isolated and prepared the platelet, then NO2- production released from aortic intima and platelet was measured with Greiss regent, NOS activity and L-arginine transport activity were detected by isotope tracer method., Results: After 7 h of WIR stress, the levels of NO2- from platelet and aortic intima obviously decreased by 57% and 46% respectively as compared with the control rats (P<0.01). The inhibitory effects of NO2- showed a positive correlation between platelet and aortic intima by person regression analysis (r2=0.9835, P<0.01). The kinetics analysis of NOS activity of platelet showed that the enzymatic catalytic maximum velocity (Vmax) decreased by 44%, Km values increase by 18% and the enzymatic catalytic efficiency was obviously reduced in the WIR stress rats (P<0.01). The aortic intima NOS activity was inhibited by 50% after WIR stress (P<0.01). The person analysis showed the obvious correlation between them (r2=0.9726, P<0.01). In the WIR stress platelet, the L-Arg transport Vmax obviously reduced by 32%, Km parameter increased 37%, and the transport efficiency was inhibited by 50% (P<0.01), which exhibited obvious positive correlation with the inhibition of L-Arg transport activity in aortic intima (27%) of WIR stress rats (r2=0.9887, P<0.01)., Conclusion: WIR stress down-regulated endogenous L-Arg/NOS/NO pathway of aortic intima and platelet, whose alteration showed obvious positive correlation. Detection of the alteration of endogenous L-Arg/NOS/NO pathway in platelet might act as an indirect method to assess the endothelial dysfunction involving the pathogensis of stress.

Purpose: To evaluate the effect of two adjuvant therapy in patients with resected maxillary mucosal melanoma., Methods: Survival analysis, retrospective study. There were 165 patients with resected maxillary mucosal malignant melanoma in the study, which were divided into three group: control group (50), interferon-alpha2b group (78) and interferon-alpha2b+dacarbazine group (37). All patients had stage II or III lesions according to AJCC(2000) staying system. Cox proportional hazards model was used to analyze the statistically significant prognostic factors. Kaplan-Meire method was used to calculate the cumulated survival rate with 3 different treatments and Log-rank method for comparison of the distribution of the different survival rates in 3 groups., Results: The overall 3-year survival rate was 4%, with 3.0% in the control group, 4.5% in interferon-<2b group and 5.7% in interferon-alpha2b+dacarbazine group, respectively. The significant prognostic factors included thickness(P<0.001, RR=2.696), ulceration(P<0.001, RR=2.068), lymph node metastasis(P<0.001, RR=1.710) and the treatment method(P<0.001, RR=0.395). A beneficial effect in overall survival was observed in two experimental groups. But the effect appeared to become weakening after 2 years. The difference between the two experimental groups was not significant statistically., Conclusions: The two conjunctive methods used in this study can improve the overall survival in patients with resected maxillary mucosal melanoma. But the beneficial effect would descend and disappear after 2 years. The effect of using interferon-alpha2b+dacarbazine was not better than that of using interferon-alpha2b alone.

Seventeen microsatellite markers of Aristichthys nobilis previously discovered by our lab were selected to analyze the genetic diversity and characteristics of two populations of Aristichthys nobilis from Jiangxi and Sichuan provinces. The following parameters were calculated: heterozygosity, polymorphism information content (PIC), valid allele number, allele frequency, genetic distance, genetic similarity coefficient, Hardy-Weinberg balance deflection index and so on. Results show that there are 4 monomorphic and 13 polymorphic markers among the 17 selected microsatellite markers. The average of allele number in each microsatellite locus of the Jiangxi population and Sichuan populations is 3.325 and 3.882, respectively; the average valid allele number is 3.531 and 2.676, respectively; and the number of total alleles of these 17 microsatellite loci is 71. The PIC of polymorphic loci varies between 0.077-0.960, and the average PIC is 0.417. The average observed heterozygosity (Ho) of two populations is 0.385 and 0.360, respectively and the average expected heterzygosity (He) is 0.452 and 0.422, respectively. The genetic similarity coefficient of two populations of Aristichthys nobilis is 0.897 and the genetic distance of these populations is 0.109.

To investigate the anti-cardiac hypertrophic mechanism of statins, thirty-eight male Wistar rats were randomly allocated to four groups. Rats in model group received nitric oxide synthase inhibitor, N-nitro-L-arginine (L-NNA) 15 mg/(kg.d) by peritoneal injection. Rats in simvastatin treatment groups were given simultaneously L-NNA as those in model group and simvastatin 5 or 30 mg/(kg.d) intragastrically respectively. Rats in control group received the same volume of normal sodium. Left ventricular function, left ventricular mass index (LVMI), the content of brain natriuretic peptide (BNP) in plasma and myocardium, myocardial hydroxyproline and heme oxygenase activity were determined after 6 weeks. The results showed that rats in model group developed significant cardiac hypertrophy associated with reduced left ventricular function compared with the control group. However, compared with the model group, L-NNA-induced cardiac hypertrophy of rats was significantly relieved in simvastatin treatment groups, associated with improved left ventricular function, decreased LVMI, lower BNP levels in plasma and myocardium, lower content of myocardial hydroxyproline, and increased myocardial heme oxygenase (HO) activity. In cultured rat neonatal cardiomyocytes, simvastatin (30 or 100 mumol/L) significantly increased heme oxygenase-1 (HO-1) mRNA expression, HO activity as well as the production of CO in cardiomyocytes. Cultured with zinc protoporphyrin, a HO inhibitor, or simvastatin alone did not change [(3)H]leucine uptake of cardiomyocytes. However, cocultured with simvastatin significantly inhibited the cardiomyocyte [(3)H]leucine uptake induced by angiotensin II in a concentration-dependent manner. Cotreatment with zinc protoporphyrin significantly abolished the suppressive effect of simvastatin on cardiomyocyte [(3)H]leucine uptake. These data suggest that the activation of HO-1/CO pathway may be one of the important mechanisms by which statins inhibit cardiac hypertrophy caused by hypertension.

Objective: To explore the change of endogenous hydrogen sulfide and the effect of exogenously applied H(2)S on Bleomycin-induced pulmonary fibrosis in rats., Methods: (1) Forty-eight male Wistar rats were randomly divided into control group, fibrosis group, fibrosis+NaHS-low group (1.4 micromol/kg, bid intraperitoneally) and fibrosis+NaHS-high group (7.0 micromol/kg, bid intraperitoneally). Pulmonary fibrosis was induced by intratracheal instillation of bleomycin. The control group and pulmonary fibrosis group were injected with the same volume of normal saline. Pulmonary pathology changes were observed, the plasma H(2)S concentration,the activity of CSE in lung tissue and the contents of MDA, hydroxyproline were detected on the 7th and 28th days respectively after bleomycin administration. CSE mRNA was also measured using quantitative RT-PCR. (2) Isolated lung tissues were exposed to Fe(2+)/H(2)O(2) without or with different concentrations of NaHS (10(-5), 5 x 10(-5), 10(-4), 10(-3) mmol/L) at 37 degree Celsius for 10 minutes, and the MDA content was assayed by biochemical method., Results: (1) Compared with rats in control group, the concentration of plasma H(2)S and the activity of CSE in lung tissues in fibrosis group decreased on day 7 (all P<0.01); On the 28th day, the level of plasma H(2)S increased clearly (P<0.01) and the activity of CSE was ascendant without significant difference compared with that of control group. Relative CSE mRNA amounts in lung tissues increased 34%, 144%, respectively (all P<0.01) in rats of fibrosis group compared with those of control group. The contents of MDA and hydroxyproline increased significantly in fibrosis group compared with those of control group on days 7, 28 (all P<0.01). In NaHS-low group and NaHS-high group, the contents of MDA and hydroxyproline decreased markedly (all P<0.01). (2) The contents of MDA in incubation of lung tissues at different concentrations of NaHS were all decreased as compared with those in incubation with Fe(2+)/H(2)O(2) alone (all P<0.01). When NaHS was at 10(-4) mol/L, the MDA content reversed to the lowest value, which had the highest value at 10(-3) mol/L of NaHS., Conclusion: Endogenous CSE/H(2)S pathway was involved in the pathogenesis of rat pulmonary fibrosis. Exogenously applied H(2)S could attenuate the process of pulmonary fibrosis possibly because of reducing oxidative stress.

Objective: To investigate the mechanism of platelet function caused by Lysophosphatidic acid (LPA), by observing the change of the L-arginine/nitric oxide synthase/nitric oxide (L-Arg/NOS/NO) pathway of platelet in rats., Methods: LPA (10(-6), 10(-5) and 5x10(-5) mol/L) was administrated in rats and incubated for 30 and 60 minutes. The nitrite production was measured by Greiss assay; NOS activities and L-arginine transportation were detected by isotope tracer method and intracellular [Ca(2+)]i changes by fluorescent probe., Results: LPA increased NO release by 35% and 56%, after incubating for 30 and 60 minutes, respectively. LPA (10(-6), 10(-5)aand 5x10(-5) mol/L) enhanced the NO productions of platelets in a concentration-dependent manner (P<0.01). EC(50) was 17.8 micromol/L, and 95% CI was 13.3-24.2 micromol/L, involved in the physiological concentration of LPA in plasma (P<0.01). Simultaneously, different doses of LPA increased NOS activities and L-arginine uptake in a dose-dependent manner (P<0.01). In this study, LPA (50 micromol/L) increased the intracellular free calcium ion concentration ([Ca(2+)]i, P<0.01), after incubating for 30 and 60 minutes. Pre-treated with NOS inhibitor-L-NAME for 20 minutes, LPA obviously enhanced the effects by 20% and 32% respectively (P<0.01). On the contrary, pre-treated with L-arginine (200 micromol/L) for the same times obviously reduced the effects by 14% and 18% respectively (P<0.01)., Conclusion: LPA increased NO release by enhancing L-arginine uptake and NOS activities, up-regulating L-arginine/NOS/NO pathway in platelets of rats.

Objective: To construct a method of measurement microamount hydrogen sulfide (H(2)S) using sensitive sulphur electrode., Methods: According to the physical and chemical characters of H(2)S, H(2)S, which in the fluids by mean of physical dissolve and chemical shape, is turned to sulphur ion (S(2-)) by chemical responses. After the microamount of S(2-) was measured by sensitive sulphur electrode, and the concentration of H(2)S was converted, a method was constructed to measure the H(2)S. It was used to analyze the concentrations of H(2)S of plasma in rats and humans, the endogenous concentration of H(2)S of cardiovascular tissue in rats, and CSE activity of cardiovascular tissues and cells., Results: The exponential regression of S(2-) in the extent including 1 to 80 micromol/L was found using sensitive sulphur electrode. The H(2)S levels of plasma in male and female rats were 40+/-4 and 41+/-5 micromol/L, respectively, and significant difference was not found; those in venous blood plasma of men and women were 33+/-4 micromol/L and 35+/-5 micromol/L respectively, without significant difference. There were not significant differences in the aortic endogenous levels of H(2)S (24+/-6 and 25+/-5 nmol/mg pro) and myocardial levels (19+/-4 and 19+/-6 nmol/mg protein) between female and male rats. There were no different results of CSE activity in aortal tissue using sensitive sulphur electrode or traditional methods, however, the CSE activity of vascular smooth muscle cells could be accurately measured using sensitive sulphur electrode, which was difficult in using traditional method., Conclusion: The sensitive sulphur electrode assay was fit for the analysis of CSE/H(2)S pathway in cardiovascular research.

Purpose: To analyze the prognostic factors which influence the survival rates in oral mucosal malignant melanoma patients., Methods: 230 consecutive patients with oral mucosal malignant melanoma were included in the study. There were 141 males and 89 females.34 were in (TNM) I stage, 87 in II stage and 109 in III stage. Cox proportional hazards model was used to analyze the statistically significant prognostic factors. Kaplan-Meier method was used to analyze the survival rate under the different levels of the different factors and Log rank method for comparison of the distribution of the different survival rates., Results: The prognostic factors include the thickness of the tumor, cervical lymph node metastasis, the presence or absence of ulceration and the anatomic sites. The P value and the relative risk were P<0.001, RR=1.868; P<0.001, RR=1.685; P<0.001, RR=1.411; P=0.008, RR=0.747, respectively. The different factors were independent with each other. The factors of sex (P=0.136), age (P=0.721) and the surgical method used (P=0.944) had no influence on survival rates in this study., Conclusions: The significant prognostic factors which influence the survival rates in oral mucosal malignant melanoma patients were similar to that in the skin malignant melanoma patients. The factors did not have factor-by-factor interactions. Surgery alone is not the best treatment of choice for oral mucosal melanoma.

Objective: To investigate the role of endogenous hydrogen sulfide (H(2)S) in patients with chronic obstructive pulmonary disease (COPD)., Methods: Levels of serum H(2)S and nitric oxide (NO), lung function and cell differential count in induced sputum were studied in 27 patients with acute exacerbation of COPD (AECOPD), 37 patients with stable COPD and 13 health subjects. Echo-Doppler assessment and arterial blood gas were measured in patients with AECOPD., Results: (1) The serum H(2)S level was significantly higher in patients with stable COPD [(50.8 +/- 2.5) micromol/L] as compared to those in the controls [(39.8 +/- 1.6) micromol/L] and in patients with AECOPD [(33.5 +/- 2.2) micromol/L, P < 0.01]. (2) The level of serum H(2)S was significantly lower in smokers with AECOPD [(28.1 +/- 1.3) micromol/L] as compared to nonsmokers with AECOPD [(39.4 +/- 3.9) micromol/L, P < 0.05] and healthy nonsmokers [(39.8 +/- 1.6) micromol/L, P < 0.01]. (3) There was significant difference in the serum H(2)S level among stable COPD patients with different severity of airway obstruction (P < 0.05); being lower in patients with stage III [(45.1 +/- 4.1) micromol/L] as compared to stage I obstruction [(70.2 +/- 6.2) micromol/L, P < 0.05]. (4) AECOPD with pulmonary hypertension pulmonary artery systolic pressure (PASP) > or = 35 mm Hg (1 mm Hg = 0.133 kPa) showed a lower serum H(2)S level [(26.3 +/- 2.2), (36.2 +/- 2.5) micromol/L, P < 0.05] than that with a normal resting PASP. (5) H(2)S in serum was positively correlated with NO levels (r = 0.278, P = 0.029), FEV(1)% predicted values (r = 0.533, P = 0.000), percentage of sputum lymphocytes (r = 0.286, P = 0.028) and macrophages (r = 0.334, P = 0.01); and negatively correlated with PASP (r = -0.561, P = 0.011) and the percentage of sputum neutrophils (r = -0.422, P = 0.001) in patients with COPD., Conclusion: Endogenous H(2)S may be involved in the pathogenesis of airway obstruction in COPD and may be a noninvasive marker of disease activity and severity.

Objective: To investigate the changes of plasma hydrogen sulfide (H(2)S) in patients with coronary heart disease (CHD)., Methods: Plasma H(2)S levels were measured in 40 patients with CHD and 17 angiographically normal patients by sulfide-sensitive electrodes, and the variation of plasma H(2)S levels was analyzed in different clinical types of CHD and in different types of coronary artery lesions. The association of plasma H(2)S levels with the risk factors of CHD was also analyzed., Results: Plasma H(2)S levels were significantly lowered in CHD patients in comparison with that in angiographically normal control subjects (26.10+/-14.27 micromol/L vs 51.74+/-11.94 micromol/L, P<0.001). In CHD patients, plasma H(2)S levels in unstable angina patients (UAP, 23.60+/-14.41 micromol/L) and acute myocardial infarction patients (AMI, 19.98+/-7.516 micromol/L) were significantly lower than that in stable angina patients (SAP, 38.41+/-14.53 micromol/L, P<0.05). No significant difference in plasma H(2)S levels was found between CHD patients with double-vessel and multi-vessel lesions (16.91+/-7.98 vs 18.39+/-7.78 micromol/L, P>0.05), but the two groups of patients had significantly lower plasma H(2)S levels than patients with single-vessel involvement (33.04+/-15.01 micromol/L, P<0.05 and P<0.01, respectively). Plasma H(2)S level was significantly lower in CHD patients with coronary artery occlusion than in patients with simple stenosis (19.04+/-9.55 vs 28.24+/-14.85 micromol/L, P<0.05). Among the CHD patients, H(2)S levels were significantly lower in smokers than in non-smokers (27.54+/-10.37 vs 32.24+/-15.77 micromol/L, P<0.05), also lower in hypertensive patients than in normotensive patients (20.36+/-8.69 vs 33.77+/-15.86 micromol/L, P<0.01). Plasma H(2)S levels showed a significant inverse correlation with blood glucose (r=-0.493 6, P=0.001 6), but there were no significant correlations with sex, age, cholesterol, triglyeride, TC, low-density lipoprotein, high-density lipoprotein, or body mass index., Conclusion: Decreased plasma H(2)S levels may correlate with the severity of CHD and changes of the coronary artery, and may implicate the risk factors of CHD such as smoking, hypertension, and high blood glucose.

Hyperhomocysteinemia is an independent risk factor for cardiovascular diseases. At present, the main therapeutic provision is to supply vitamin B(6), B(12) and folic acids, and the toxic and ill effect have been reported. Homocysteine, taurine, hydrogen sulfide and metallothionein are metabolic products from methionine. Homocysteine induces necrosis of endothelium, proliferation of vascular smooth muscle cells, proliferation and activation of vascular fibroblast cells and mitochondrial structural destruction and dysfunction of myocardium cells. Taurine, an end metabolic product of homocysteine, obviously reduces cardiovascular injury induced by homocysteine. The possible mechanism of antagonism by reducing oxidative stress and endoplasmic reticulum stress has been proved. Hydrogen sulfide, another end metabolic product of homocysteine, obviously reducing cardiovascular injury of homocysteine by scavenging oxidative radicals has been found. Metallothionein a derivant production of homocysteine metabolism, antagonism to homocysteine injury to cardiovascular system been discussed. Homocysteine, taurine, hydrogen sulfide and metallothionein, as a metabolic product of methionine, interactive antagonism and interactive biological influence have been reviewed. Induced endogenous or exogenous supply of taurine, hydrogen sulfide and metallothionein might resist cardiovascular injury induced by hyperhomocysteinemia. According to the methionine metabolic cycle, using endogenous antagonistic substances might be a new clinical preventive and treatment target of homocystinemia.

Hydrogen sulfide (H(2)S) may be endogenously produced by cystathionine beta-lyase (CBS) and cystathionine gamma-lyase (CSE) as a cardiovascular physiological functional factor. On the hypoxic pulmonary hypertension (HPH) animal model, the plasma H(2)S concentration, the gene expression and the activity (CSE) were decreased in lung tissues In L-NAME induced hypertension and spontaneous hypertension rats (SHR) models, the plasma H(2)S concentration, vascular CSE activity and mRNA expression were obviously decreased. When H(2)S was exogenously supplied, systolic pressure obviously decrease. These studies suggested that CSE/H(2)S pathway participated in the pathophysiological development of hypertension. The endogenous level of H(2)S produced by some arterial tissues increased in both septic and endotoxic shock rats. The level of H(2)S highly correlated with the endogenous level of NO. These results suggest that H(2)S may be a novel cardiovascular functional regulator.

Objective: To study the effects of adrenomedullin (ADM) and proadrenomedullin N-terminal 20 peptide (PAMP) alone or in combinations on the isolated rat hearts as well as the possible signaling pathways involved in their actions., Methods: In isolated rat hearts the left ventricular pressure (LVP), LVP+/-dp/dtmax, coronary fluid (CF) and heart rate(HR) of the hearts infused at different concentrations of ADM and/or PAMP were determined by a 4-cannal physiological recorder, then the cAMP contents were assayed in myocardium., Results: After being infused with ADM from 10(-11) to 10(-8) mol x L(-1) or PAMP from 10(-11) to 10(-8)mol x L(-1), the LVP and LVP+/-dp/dtmax of the isolated hearts decreased gradually in a concentration-dependent manner, and at the same concentration, the effects of PAMP were more potent than those of the ADM. When ADM and PAMP were co-administrated with both concentrations as low as from 10(-11) to 10(-10) mol x L(-1), the cardiac parameters were decreased more than either ADM or PAMP administrated alone. However, the inhibitory effects of ADM and PAMP were attenuated when they were in combination at higher concentrations as from 10(-9) to 10(-8) mol x L(-1). When the rat hearts were infused with ADM PAMP and ADM plus PAMP, the CF were always higher than those of the controls and decreased when co-administrated with L-NAME, an inhibitor of NOS, but the decreased degree of LVP and LVP+/-dp/dtmax were attenuated by L-NAME. The cAMP contents in the left cardiac ventricle were increased significantly by ADM infusions but not changed obviously by PAMP, and were of no statistical difference in rat hearts with ADM administrated alone from those combinated with ADM and PAMP., Conclusion: These results showed that ADM and PAMP infused alone or in combinations inhibited the function of rat hearts in vitro, which might be partly involved with the NOS/NO pathway.