从“旧”厂房改造活化到“新”科研高精工艺实验室

科研实验室的设计规划,无论是新建、扩建还是改建工程,是一项复杂的系统工程;不仅是实验室的整体规划、合理布局和平面设计,还涉及供电、供水、供气、通风、空气净化、废物清运、环境保护(如涉及核素、X射线等)等基本条件;同时实验室设备基础需求、科研设备用电需求、洁净室需求、运输通道、装饰装修等也密切相关。通过光明科学城重大科研项目过渡场地建设工程的实践,阐述了光明科研实验室项目建设过程中从“旧”厂房改造活化到“新”科研高精工艺实验室中“人”的需求和“物”的需求的实验环境及技术措施,为国内同类旧改实验室的建设提供经验。

NaOH活化木纤维表面形态及结构的分析

为了研究NaOH活化条件对木纤维表面形态及结构的影响,采用扫描电镜、红外光谱和X-射线衍射观察和分析不同NaOH用量、活化时间和活化温度处理后木纤维表面形态、羟基数量及纤维素结晶度的变化。结果表明:经NaOH活化的纤维表面不再光滑,出现分层、凹陷和空洞;活化条件对纤维表面羟基数量和纤维素结晶度的影响趋势不一致;当NaOH用量6%、180℃活化30min时,经红外测得羟基的吸收峰最强;当NaOH用量6%、180℃活化15min时,纤维素结晶度由40.80%增至43.23%,增幅最大。

Regeneration of activated carbon adsorbed EDTA by electrochemical method

Activated carbon after saturated adsorption of EDTA was used as particle electrode in a three-dimensional electrode reactor to treat EDTA-containing wastewater.Electrochemical method was used to regenerate activated carbon after many times of electrolysis.Based on the analysis of infrared spectra of activated carbon after adsorption and repeated electrolysis,EDTA was degraded into glycine,and then non-catalytic activated associated complex was formed with N—H bond on the activated carbon.The catalytic ability of the activated carbon vanished and the EDTA degradation efficiency was dropped.Activated carbon could be effectively regenerated by electrochemical method in the three-dimensional reactor.Effects of electric current,conductivity and pH on activated carbon regeneration were investigated,and the optimum conditions were concluded as follows：100-300 mA of current intensity,1.39 mS/cm of electric conductivity,60 min of electrolysis time and pH 6.0-8.0.Under the optimized conditions,the activity of the activated carbon can be recovered and the residual total organic carbon（TOC） was below 10 mg/L（the initial TOC was 200 mg/L） in the three-dimensional electrode reactor.

Synthesis of one-dimensional carbon nanostructures and their application as anode materials in lithium ion batteries

An efficient synthesis of carbon nanofibers by pyrolysis of as-prepared polypyrrole nanowires was reported. Under the subsequent KOH activation, a significant morphology variation was detected and the obtained sample took on a ribbon-like structure. The morphology and structure of the carbon nanofibers and carbon nanoribbons were characterized. When the as-prepared one-dimensional carbon nanostructures were used as anode materials in lithium ion batteries, both of them exhibited superior cyclical stability and good rate properties. After 50 cycles, the reversible capacity of carbon nanofibers electrode maintained 530 mA·h/g. Concerning carbon nanoribbons, the reversible capacity is always larger than 850 mA·h/g and the reversible capacity retention after 23 cycles is 86%.

Modification of ACFs by chemical vapor deposition and its application for removal of methyl orange from aqueous solution

Viscose activated carbon fibers （ACFs） were characterized using specific surface area, scanning electron modified with chemical vapor deposition （CVD）. The samples were microscopy （SEM）, pore size distribution and Fourier transform infrared spectroscopy （FTIR）. Batch adsorption experiments were carried out to investigate the adsorption behavior of modified ACFs for methyl orange（MO） from its aqueous solutions. The results show that the adsorption isotherms of MO onto modified ACFs well follows the Langmuir isotherm equation. The adsorption kinetics of MO can be well described by the pseudo second-order kinetic model. The adsorption process involves the intra-particle diffusion, but is not the only rate-controlling step. Thermodynamic parameters including AG, AH and AS were calculated, suggesting that the adsorption of MO onto modified ACFs is a spontaneous, exothermic and physisorption process. FTIR result indicates that the major adsorption mechanism of modified ACFs for MO is hydrogen bond.

Notch3 regulates the activation of hepatic stellate cells

AIM： To investigate whether Notch signaling is involved in liver fibrosis by regulating the activation of hepatic stellate cells （HSCs）.METHODS： Immunohistochemistry was used to detect the expression of Notch3 in fibrotic liver tissues of patients with chronic active hepatitis. The expression of Notch3 in HSC-T6 cells treated or not with transforming growth factor （TGF）-β1 was analyzed by iramunofluorescence staining, The expression of Notch3 and myofibroblastic marker （z-smooth muscle actin （（z-SMA） and collagen 1 in HSC-T6 cells transfected with pcDNA3. I-N3ICD or control vector were detected by Western blotting and immunofluorescence staining. Moreover, effects of Notch3 knockdown in HSC-T6 by Notch3 siRNA were investigated by Western blotting and immunofluorescence staining.RESULTS： The expression of Notch3 was significantly up-regulated in fibrotic liver tissues of patients withchronic active hepatitis, but not detected in normal liver tissues. Active Notch signaling was found in HSC-T6 cells. TGF-β1 treatment led to up-regulation of Notch3 expression in HSC-T6 cells, and over-expression of Notch3 increased the expression of α-SMA and collagen I in HSC-T6 without TGF-β1 treatment. Interestingly, transient knockdown of Notch3 decreased the expression of myofibroblastic marker and antagonized TGF-β1 induced expression of α-SMA and collagen I in HSC-T6.CONCLUSION： Notch3 may regulate the activation of HSCs, and the selective interruption of Notch3 may provide an anti -fibrotic strategy in hepatic fibrosis.

Fibrous activated alumina prepared through phase transformation using dawsonite as a template

Fibrous activated alumina is widely applied in catalysts,adsorbents,and composite materials.This work presents a green approach in preparing the fibrous activated Al_(2)O_(3) with high purity and specific surface area through multistep phase transformation of aluminum-bearing substances using intermediate dawsonite as a template.Thermodynamic calculations and experimental results show that increasing the concentration of Na_(2)CO_(3) and(NH_(4))_(2)CO_(3) is remarkably beneficial to the formation of dawsonite and ammonium aluminum carbonate hydroxide,respectively.Based on determination of dissolution and precipitation mechanism,the ultrafine granular gibbsite is converted to the uniform fibrous dawsonite with a ratio of length to diameter over 50,and the fibrous dawsonite changes into the long fibrous ammonium aluminum carbonate hydroxide with a ratio of length to diameter is about 80 in above 70 g/L(NH_(4))_(2)CO_(3) solution.Furthermore,the activated alumina remains fibrous morphology after roasting ammonium aluminum carbonate hydroxide at a slow heating rate,plentiful open mesopore and weak aggregation of particles,which contributes to the high specific surface area of 159.37 m^(2)/g at 1273 K for the activated alumina.The complete transformation of dawsonite to ammonium aluminum carbonate hydroxide and high specific surface area contribute to the purity of the activated fibrous alumina above 99.9%with low Na and Fe content.

Ultrastructure of oval cells in children with chronic hepatitis B,with special emphasis on the stage of liver fibrosis:The first pediatric study

AIM:To investigate the ultrastructure of oval cells in children with chronic hepatitis B,with special emphasis on their location in areas of collagen fibroplasia. METHODS:Morphological investigations were conducted on biopsy material obtained from 40 children,aged 3-16 years with chronic hepatitis B. The stage of fibrosis was assessed histologically using the arbitrary semiquantitative numerical scoring system proposed by Ishak et al. The material for ultrastructural investigation was fixed in glutaraldehyde and paraformaldehyde and processed for transmission-electron microscopic analysis. RESULTS:Ultrastructural examination of biopsy specimens obtained from children with chronic hepatitis B showed the presence of two types of oval cells,the hepatic progenitor cells and intermediate hepatic-like cells. These cells were present in the parenchyma and were seen most commonly in areas of intense periportal fibrosis (at least stage 2 according to Ishak et al) and in the vicinity of the limiting plate of the lobule. The activated nonparenchymal hepatic cells,i.e. transformed hepatic stellate cells and Kupffer cells were seen in close proximity to the intermediate hepatic-like cells. CONCLUSION:We found a distinct relationship between the prevalence of oval cells (hepatic progenitor cells and intermediate hepatocyte-like cells) and fibrosis stage in pediatric patients with chronic hepatitis B.

Lipid accumulation in hepatocytes induces fibrogenic activation of hepatic stellate cells

Despite the initial belief that non-alcoholic fatty liver disease is a benign disorder, it is now recognized that fibrosis progression occurs in a significant number of patients. Furthermore, hepatic steatosis has been identified as a risk factor for the progression of hepatic fibrosis in a wide range of other liver diseases. Here, we established an in vitro model to study the effect of hepatic lipid accumulation on hepatic stellate cells （HSCs）, the central mediators of liver fibrogenesis. Primary human hepatocytes were incubated with the saturated fatty acid palmitate to induce intracellular lipid accumulation. Subsequently, human HSCs were incubated with conditioned media （CM） from steatotic or control hepatocytes. Lipid accumulation in hepatocytes induced the release of factors that accelerated the activation and proliferation of HSC, and enhanced their resistance to apoptosis, largely mediated via activation of the PI-3-kinase pathway. Furthermore, CM from steatotic hepatocytes induced the expression of the profibrogenic genes TGF-β, tissue inhibitor of metallo-proteinase-1 （TIMP-1）, TIMP-2 and matrix-metallo-proteinase-2, as well as nuclear-factor κB-dependent MCP-1 expression in HSC. In summary, our in vitro data indicate a potential mechanism for the pathophysiological link between hepatic steatosis and fibrogenesis in vivo. Herewith, this study provides an attractive in vitro model to study the molecular mechanisms of steatosis-induced fibrogenesis, and to identify and test novel targets for antifibrotic therapies in fatty liver disease.

A red wine polyphenolic extract reduces the activation phenotype of cultured human liver myofibroblasts

AIM: To test the effect of a standardized red wine polyphenolic extract (RWPE) on the phenotype of human liver myofibroblasts in culture. METHODS: Human myofibroblasts grown from liver explants were used in this study. Cell proliferation was measured with the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay. Signaling events were analyzed by western blot with phosphospecific antibodies. Matrix-metalloproteinase activity was measured with gel zymography. RESULTS: We found that cell proliferation was dose-dependently decreased by up to 90% by RWPE while cell viability was not affected. Exposure to RWPE also greatly decreased the phosphorylation of ERK1/ERK2 and Akt in response to stimulation by the mitogenic factor platelet-derived growth factor BB (PDGF-BB). Finally, RWPE affected extracellular matrix remodeling by decreasing the secretion by myofibroblasts of matrix-metalloproteinase-2 and of tissue inhibitor of matrix- metalloproteinases-1.CONCLUSION: Altogether, RWPE decreases the activation state of liver myofibroblasts. The identification of the active compounds in RWPE could offer new therapeutic strategies against liver fibrosis.

Is liver biopsy mandatory in children with chronic hepatitis C?

Liver biopsy is considered the most accurate means to estimate the necroinflammatory activity and the extent of fibrosis. However, histology evaluation is an invasive procedure associated with risk to the patient, risk of sampling error and diagnostic inconsistencies due to inter- and intra-observer error. On the basis of histological studies performed so far, chronic hepatitis C in children appears morphologically benign in the majority of cases. At the Pediatric Liver Unit of our university, a total of 67 children with chronic hepatitis C underwent liver biopsy. Liver biopsy was repeated 5.5 years after the initial histological evaluation in 21 children. On a total number of 88 liver biopsies, micronodular cirrhosis was detected only in one genotype 1b-infected obese child. Since liver histology investigation of a child with chronic hepatitis C has few chances to highlight severe lesions, we question how liver biopsy helps in the management of children with chronic hepatitis C.

Why,who and how should perform liver biopsy in chronic liver diseases

Chronic viral hepatitis is a common disease in the general population.During chronic hepatitis,the prognosis and clinical management are highly dependent on the extent of liver fibrosis.The fibrosis evaluation can be performed by FibroTest(using serological markers),by Elastography or FibroScan(a noninvasive percutaneous technique using the elastic properties of the hepatic tissue) and by liver biopsy(LB),considered to be the "gold standard".Currently,there are three techniques for performing LB:percutaneous,transjugular and laparoscopic.The percutaneous LB can be performed blind,ultrasound(US) guided or US assisted.There are two main categories of specialists who perform LB:gastroenterologists(hepatologists) and radiologists,and the specialty of the individual who performs the LB determines if the LB is performed under ultrasound guidance or not.There are two types of biopsy needles used for LB:cutting needles(Tru-Cut,Vim-Silverman) and suction needles(Menghini,Klatzkin,Jamshidi).The rate of major complications after percutaneous LB ranges from 0.09% to 2.3%,but the echo-guided percutaneous liver biopsy is a safe method for the diagnosis of chronic diffuse hepatitis(cost-effective as compared to blind biopsy) and the rate of complications seems to be related to the experience of the physician and the type of the needle used(Menghini type needle seems to be safer).Maybe,in a few years we will use non-invasive markers of fibrosis,but at this time,most authorities in the field consider that the LB is useful and necessary for the evaluation of chronic hepatopathies,despite the fact that it is not a perfect test.

Differential gene expression during capillary morphogenesis in a microcarrier-based three-dimensional in vitro model of angiogenesis with focus on chemokines and chemokine receptors

AIM:To globally compare the gene expression profiles during the capillary morphogenesis of human microvascular endothelial cells (HMVECs) in an in vitro angiogeness system with affymetrix oligonucleotide array. METHODS: A microcarrier-based in vitro angiogenesis system was developed, in which ECs migrated into the matrix, proliferated, and formed capillary sprouts. The sprouts elongated, branched and formed networks. The total RNA samples from the HMVECs at the selected time points (0.5, 24, and 72 h) during the capillary morphogenesis were used for microarray analyses, and the data were processed with the softwares provided by the manufacturers. The expression patterns of some genes were validated and confirmed by semi-quantitative RT-PCR. The regulated genes were grouped based on their molecular functions and expression patterns, and among them the expression of chemokines and chemokine receptors was specially examined and their functional implications were analyzed. RESULTS: A total of 1 961 genes were up- or downreg-ulated two-folds or above, and among them, 468 genes were up- or down-regulated three-folds or above. The regulated genes could be grouped into categories based on their molecular functions, and were also clustered into six groups based on their patterns of expression. As for chemokines and chemokine receptors, CXCL1/GRO-α, CXCL2/GRO-β, CXCL5/ENA-78, CXCL6/GCP2, IL-8/CXCL8, CXCL12/SDF-1, CXCL9/Mig, CXC11/ITAC, OOCL1/fractalkine, CCL2/MCP-1, CCL3, CCL5/RANTES, CCL7, CCL15, CCL21, CCL23, CCL28, and CCR1, CCR9, CXCR4 were identified. Moreover, these genes demonstrated different changing patterns during the capillary morphogenesis, which implied that they might have different roles in the sequential process. Among the chemokines identified, CCL2/MCP-1, CCL5/RANTES and CX3CL1 were specially up-regulated at the 24-h time point when the sprouting characterized the morphological change. It was thus suggested that they might exert crucial roles at the early stage of angiogenesis. CONCLUSION: The present study demonstrates a global profile of gene expression during endothelial capillary morphogenesis, and the results provide us much information about the molecular mechanisms of angiogenesis, with which further evaluation of individual genes can be conducted.

Non invasive fibrosis biomarkers reduce but not substitute the need for liver biopsy

Chronic liver diseases are very common worldwide, particularly those linked to viral hepatitis and to alcoholic and non-alcoholic fatty liver. Their natural history is variable and long-term evolution differs in individual patients. Optimised clinical management of compensated chronic liver diseases requires precise definition of the stage of liver fibrosis, the main determinant of prognosis and of most therapeutic decisions. Liver biopsy is the gold standard for assessment of hepatic fibrosis. However, it is invasive with possible complications, costly and prone to sampling errors. Many non-invasive markers of liver fibrosis have been recently proposed and assessed in the clinical setting as surrogates of liver biopsy. Direct markers are based on biochemical parameters directly linked to fibrogenesis while indirect markers use simple or more sophisticated parameters that correlate with liver fibrosis stages. Non-invasive markers of liver fibrosis have been tested in different forms of chronic liver disease and showed variable diagnostic performance, but accuracy rarely was above 75%-80%. Better results were obtained when markers were combined. On this line, we have recently proposed a set of algorithms that combine sequentially indirect non-invasive markers of liver fibrosis, reaching 90%-95% diagnostic accuracy with significant reduction in the need for liver biopsy. Based on available evidence, it can be anticipated that non-invasive markers of liver fibrosis and their combined use will soon become a most useful tool in the clinical management of many forms of chronic liver disease. However, their implementation is expected to reduce, but not to completely eliminate, the need for liver biopsy.

Preparation of activated carbon with low ash content and high specific surface area from coal in the presence of KOH

An activated carbon with ash content less than 10% and specific surface area more than 1 600 m 2 /g was prepared from coal and the effect of K containing compounds in preparation of coal based activated carbon was investigated in detail in this paper. KOH was used in co carbonization with coal, changes in graphitic crystallites in chars derived from carbonization of coal with and without KOH were analyzed by X ray diffraction (XRD) technique, activation rates of chars with different contents of K containing compounds were deduced, and resulting activated carbons were characterized by nitrogen adsorption isotherms at 77 K and iodine numbers. The results showed that the addition of KOH to the coal before carbonization can realize the intensive removal of inorganic matters from chars under mild conditions, especially the efficient removal of dispersive quartz, an extremely difficult separated mineral component in other processes else. Apart from this, KOH demonstrates a favorable effect in control over coal carbonization with the goal to form nongraphitizable isotropic carbon precursor, which is a necessary prerequisite for the formation and development of micro pores. However, the K containing compounds such as K 2 CO 3 and K 2 O remaining in chars after carbonization catalyze the reaction between carbon and steam in activation, which leads to the formation of macro pores. In the end an innovative method, in which KOH is added to coal before carbonization and K containing compounds are removed by acid washing after carbonization, was proposed for the synthesis of quality coal based activated carbon.

Significant roles of anti-aging protein klotho and fibroblast growth factor23 in cardiovascular disease

The klotho gene has been identified as an aging suppressor that encodes a protein involved in cardiovascular disease （CVD）. The inac- tivation of the klotho gene causes serious systemic disorders resembling human aging, such as atherosderosis, diffuse vascular calcification and shortened life span. Klotho has been demonstrated to ameliorate vascular endothelial dysfunction and delay vascular calcification. Fur- thermore, klotho gene polymorphisms in the human are associated with various cardiovascular events. Recent experiments show that klotho may reduce transient receptor potential canonical6 （TRPC6） channels, resulting in protecting the heart from hypertrophy and systolic dys- function. Fibroblast growth factor23 （FGF23） is a bone-derived hormone that plays an important role in the regulation of phosphate and vi- tamin D metabolism. FGF23 accelerates urinary phosphate excretion and suppresses 1,25-dihydroxy vitaminD3 （1,25（OH）2D3）synthesis in the presence ofFGF receptorl （FGFR1） and its co-receptor ldotho, principally in the kidney. The hormonal affects of circulating klotho pro- tein and FGF23 on vascular and heart have contributed to an understanding of their roles in the pathophysiology of arterial stiffness and left ventricular hypertrophy. Klotho and FGF23 appear to play a critical role in the pathogenesis of vascular disease, and may represent a novel potential therapeutic strategy for clinical intervention.

Ceruloplasmin, a reliable marker of fibrosis in chronic hepatitis B virus patients with normal or minimally raised alanine aminotransferase

AIM To develop a non-invasive model to evaluate significant fibrosis and cirrhosis by investigating the association between serum ceruloplasmin(CP) levels and liver fibrosis in chronic hepatitis B(CHB) patients with normal or minimally raised alanine aminotransferase(ALT).METHODS Serum samples and liver biopsy were obtained from 193 CHB patients with minimally raised or normal ALT who were randomly divided into a training group(n = 97) and a validation group(n = 96). Liver histology was evaluated by the METAVIR scoring system. Receiver operator characteristic curves were applied to the diagnostic value of CP for measuring liver fibrosis in CHB patients. Spearman rank correlation analyzed the relationship between CP and liver fibrosis. A noninvasive model was set up through multivariate logistic regression analysis.RESULTS Serum CP levels individualized various fibrosis stages via area under the curve(AUC) values. Multivariate analysis revealed that CP levels were significantlyrelated to liver cirrhosis. Combining CP with serum GGT levels, a CG model was set up to predict significant fibrosis and liver cirrhosis in CHB patients with normal or minimally raised ALT. The AUC, sensitivity, specificity, positive predictive value, and negative predictive value were 0.84, 83.1%, 78.6%, 39.6%, and 96.5% to predict liver cirrhosis, and 0.789, 80.26%, 68.38%, 62.25%, and 84.21% to predict significant fibrosis. This model expressed a higher AUC than FIB-4(age, ALT, aspartate aminotransferase, platelets) and GP(globulin, platelets) models to predict significant fibrosis(P = 0.019 and 0.022 respectively) and revealed a dramatically greater AUC than FIB-4(P = 0.033) to predict liver cirrhosis.CONCLUSION The present study showed that CP was independently and negatively associated with liver fibrosis. Furthermore, we developed a novel promising model(CG), based on routine serum markers, for predicting liver fibrosis in CHB patients with normal or minimally raised ALT.

Optimization of Culture Conditions for Production of Cellulase by a Thermophilic Bacillus Strain

The production of cellulase in Bacillus amyloliquefaciens UNPDV-22 was optimized using response surface methodology （RSM）. Central composite design （CCD） was used to study the interactive effect of culture conditions （temperature, pH, and inoculum） on cellulase activity. Results suggested that temperature and pH all have significant impact on cellulase production. The use of RSM resulted in a 96% increase in the cellulase activity over the control of non-optimized basal medium. Optimum cellulase production of 13 U/mL was obtained at a temperature of 42.24 ℃, pH of 5.25, and inoculum size of 4.95% （v/v） in a fermentation medium containing wheat bran, soybean meal and malt dextrin as major nutritional factors.

Effect of dietary supplementation of vitamin C on growth, reactive oxygen species, and antioxidant enzyme activity of Apostichopus japonicus(Selenka) juveniles exposed to nitrite

Different amounts of vitamin C were added to diets fed to juveniles （2.5＋0.15 g） of sea cucumber Apostichopus japonicus （Selenka） in an attempt to reduce the stress response of specimens exposed to nitrite stress. A commercial feed was used as the control diet and three experimental diets were made by supplementing 1 000, 1 500, or 2 000 mg vitamin C/kg diet to control diet separately in a 45-day experiment. Sea cucumbers were exposed to three different levels （0.5, 1.0, and 1.5 mg/L） of nitrite stress for 4, 8, and 12 h at four time intervals （0, 15, 30, and 45 d）. Growth of the animals was recorded during the experiment. Reactive oxygen species （ROS） （i.e. hydroxyl free radical （-OH）, malondialdehyde （MDA） and total antioxidant capacity （T-AOC）） and antioxidant enzyme activities （i.e., superoxide dismutase （SOD） and eatalase （CAT）） were measured. Response surface methodology （RSM） was used to analyze the effect of multiple factors on ROS indices and enzyme activities. Weight gain （WG） and special growth rate （SGR） of vitamin C supplementation groups were significantly higher than those of control group （P〈0.05）. The levels of-OH and MDA increased under exposure time extending and nitrite concentration increasing, whereas T-AOC level decreased. SOD and CAT activities increased at 4 h and 8 h and decreased at 12 h. During the days in which the animal consumed experimental diets, the levels of-OH and MDA decreased and that of T-AOC increased. This result suggests that diets containing vitamin C could reduce the nitrite stress response in the animals and increase their antioxidant capacity. The multifactor regression equation of growth performance, ROS indices, and duration of feeding results suggest that vitamin C supplementation of 1 400-2 000 mg/kg diet for 29-35 days could reduce effectively the effects of nitrite exposure.

Characterization of Crude Cellulase from Trichoderma reesei and Purification of Cellulase

The gel filtration was carried out for purification of cellulase. The influences of chromatographic parameters on the resolution were studied to determine the optimal conditions for purification. The purified endoglucanase was obtained by gel filtration by Superdex 75 prep grade with an activity recovery of 92.8% and the purification factor 4.2. The sample volume should be below 6% of the column bed volume and the column bed height L≥12.0 cm. The optimum catalysis temperature and pH for the enzyme were 55 ℃ and 4.5—5.0, respectively. The cellulase was stable at pH ranging from 4.0 to 6.0 and temperature below 60 ℃.