Rice straw decomposition in rice-field soil

Rice straw, buried in a rice-field during the dry season decomposed at a rate of 0.0075 day-1. Seventy five percent of the biomass, 70 percent carbon, 50 percent nitrogen and 30 percent phosphorus remained after 139 days of decomposition. Rice straw decomposition furnished 33% N and 8% P of the total nitrogen and phosphorus provided by man.     

Organic Matter Dynamics in a Tropical Gallery Forest in a Grassland Landscape

The high biodiversity of tropical forest streams depends on the strong input of organic matter, yet the leaf litter decomposition dynamics in these streams are not well understood. We assessed how seasonal litterfall affects leaf litter breakdown, density and biomass of aquatic invertebrates, and the microbial biomass and sporulation of aquatic hyphomycetes in a South American grassland ‘vereda’ landscape. Although litter production in the riparian area was low, leaf litter breakdown was high compared with other South American systems, with maximum values coinciding with the rainy season. Fungal biomass in decomposing leaves was high, but spore densities in water and sporulation rates were very low. Invertebrates were not abundant in litter bags, suggesting they play a minor role in leaf litter decomposition. Chironomids accounted for ~70 percent of all invertebrates; only 10 percent of non‐Chironomidae invertebrates were shredders. Therefore, fungi appear to be the drivers of leaf litter decomposition. Our results show that despite low productivity and relatively fast litter decomposition, organic matter accumulated in the stream and riparian area. This pattern was attributed to the wet/dry cycles in which leaves falling in the flat riparian zone remain undecomposed (during the dry period) and are massively transported to the riverbed (rainy season).     

Control of ovarian cholesterol ester biosynthesis

1. Experimental evidence is presented for a role of progesterone and 20alpha-hydroxypregn-4-en-3-one as inhibitors of cholesterol ester synthetase in the acute depletion of ovarian cholesterol ester after trophic stimulation. 2. Luteinizing hormone in vitro decreased by 84% the rate of esterification of cholesterol with added [(14)C]oleate by slices of rabbit ovarian interstitial tissue; this effect was mimicked by cyclic AMP (adenosine 3':5'-cyclic monophosphate) in vitro, and occurred without large changes in precursor pool sizes or membrane permeability. 3. Cyclic AMP was shown to have no direct effect on cholesterol ester synthetase or cholesterol esterase in cell-free extracts of rabbit ovarian interstitial tissue, but decreased the activity of cholesterol ester synthetase (not that of cholesterol esterase) in extracts prepared from slices previously incubated with it. 4. The inhibitory effect of cyclic AMP on esterification of cholesterol with added [(14)C]-oleate was prevented by both cycloheximide and aminoglutethimide phosphate (which also inhibited steroid synthesis in response to cyclic AMP). 5. Cyclic AMP raised the intracellular concentrations of progesterone and 20alpha-hydroxypregn-4-en-3-one in incubated slices by factors of 2.8 and 3.9 respectively. 6. Cycloheximide and aminoglutethimide phosphate administered in vivo blocked cholesterol ester depletion in response to luteinizing hormone in rats; in these ovaries cycloheximide and aminoglutethimide phosphate decreased the concentrations of progesterone and 20alpha-hydroxypregn-4-en-3-one and luteinizing hormone raised them. 7. Progesterone and 20alpha-hydroxypregn-4-en-3-one added to cell-free extracts of rabbit ovarian interstitial tissue in vitro (at concentrations comparable with those found in incubated slices) inhibited cholesterol ester synthetase by up to 85%. 8. The results are discussed with reference to the acute control of cholesterol ester concentrations in the ovary and adrenal cortex.     

罗氏乳杆菌无细胞上清培养液移除胆固醇能力的研究

本文探讨了罗氏乳杆菌DSM122460无细胞上清培养液(Cell-Free Supernatant,CFS)移除胆固醇的能力。采用邻苯二甲醛法测定DSM122460和对照菌株ST-III发酵过程中及其CFS对胆固醇的移除能力,并研究不同CFS浓度下的移除能力。并采用HPLC法测定CFS对照、热处理组和pH7.0组的胆盐水解酶活力,同时测定其移除胆固醇能力。结果显示,DSM122460不仅在发酵过程中具有较高的移除胆固醇能力,其CFS也表现出较高的移除能力,CFS中含有除胆盐水解酶以外的可移除胆固醇的蛋白类成分。这提示可能存在一种乳酸菌移除胆固醇的新机制。     

Hydroperoxysterols as a probe for the mechanism of cytochrome P-450scc-mediated hydroxylation. Homolytic versus heterolytic oxygen-oxygen bond scission

The interaction of 20-, 23-, and 25-hydroperoxy derivatives of cholesterol with various heme proteins, including the cholesterol side-chain-cleaving enzyme, cytochrome P-450scc, was studied by means of product and spectral analyses. Quasi-Fenton homolytic decomposition via intermediate alkoxy radicals appears to prevail during nonspecific interaction. Highly stereospecific hydroperoxide-driven hydroxylations suggest the absence of free radical species and are interpreted as resulting from a heterolytic type of peroxide decomposition or, alternatively, homolytic decomposition assuming proximal base effect to stabilize a putative intermediate alkoxy radical. Spectral aberrations during the early stages of the latter interaction indicate formation of a ternary iron-peroxo-substrate complex. Decomposition of this complex results in multiple product formation suggesting that peroxide cleavage reverts to regular homolytic decomposition upon denaturation of the enzyme. The implications of these observations for the mechanism of hydroxylation and oxidative carbon-carbon bond scission during enzymic side-chain cleavage of cholesterol are discussed.     

Towards a budget of leaf litter decomposition in a first-order woodland stream

To construct a budget of carbon transformations occurring during leaf decomposition, alder leaves were placed in a woodland stream, later retrieved at weekly intervals, and rates of fungal and bacterial production, microbial respiration, and release of dissolved organic matter (DOM) and fine particulate organic matter (FPOM) were determined during short laboratory incubations. Carbon dioxide was the major decomposition product, explaining 17% of the microbially mediated leaf mass loss. DOM and FPOM were also important products (5 and 3% of total mass loss, respectively), whereas carbon flow to microbial biomass was low (2%). Fungal biomass in leaves always exceeded bacterial biomass (95–99% of total microbial biomass), but production of bacteria and fungi was similar, indicating that both types of microorganisms need to be considered when examining leaf decomposition in streams. Comparison of leaf mass loss in coarse and fine mesh bags revealed, in addition, that the shredder, Gammarus pulex, had a major impact on leaf decomposition in this study.     

Organic matter availability structures microbial biomass and activity in a Mediterranean stream

1. We compared microbial biomass (bacteria, fungi, algae) and the activity of extracellular enzymes used in the decomposition of organic matter (OM) among different benthic substrata (leaves, coarse and fine substrata) over one hydrological year in a Mediterranean stream.
2. Microbial heterotrophic biomass (bacteria plus fungi) was generally higher than autotrophic biomass (algae), except during short periods of high light availability in the spring and winter. During these periods, sources of OM shifted towards autochthonous sources derived mainly from algae, which was demonstrated by high algal biomass and peptidase activity in benthic communities.
3. Heterotrophic activity peaked in the autumn. Bacterial and fungal biomass increased with the decomposition of cellulose and hemicellulose compounds from leaf material. Later, lignin decomposition was stimulated in fine (sand, gravel) and coarse (rocks, boulders and cobbles) substrata by the accumulation of fine detritus.
4. The Mediterranean summer drought provoked an earlier leaf fall. The resumption of the water flow caused the weathering of riparian soils and subsequently a large increase in dissolved organic carbon and nitrate, which led to growth of bacteria and fungi.     

长白山次生针阔混交林叶凋落物中有机物分解与碳、氮和磷释放的关系

采用分解袋法,研究了长白山次生针阔混交林内9种树种叶凋落物34个月的分解过程.结果表明:在次生针阔混交林中,不同树种叶凋落物的初始N和P浓度存在很大差异,叶凋落物分解速率(k)与初始N浓度和C/N显著相关.有机物剩余百分率与C剩余百分率呈显著的线性正相关关系,与N和P剩余百分率之间呈显著的二项式回归关系. N和P剩余百分率在初期阶段随有机物分解而增加,达到峰值后逐渐降低. 随着有机物剩余百分率的下降,C/N和C/P均呈逐渐降低趋势,各种叶凋落物之间C/N和C/P的差异逐渐减小,分解末期分别趋近于23和350. 随有机物剩余百分率的降低, N/P变化不明显,当有机物剩余百分率低于25%时,不同树种叶凋落物之间N/P差异显著下降. 有机物剩余百分率可用来预测C、N和P的剩余百分率.      

Changes in leaf litter decomposition of primary Korean pine forests after degradation succession into secondary broad‐leaved forests

Forest degradation succession often leads to changes in forest ecosystem functioning. Exactly how the decomposition of leaf litter is affected in a disturbed forest remains unknown. Therefore, in our study, we selected a primary Korean pine forest (PK) and a secondary broad‐leaved forest (SF) affected by clear‐cutting degradation, both in Northeast China. The aim was to explore the response to changes in the leaf litter decomposition converting PK to SF. The mixed litters of PK and SF were decomposed in situ (1 year). The proportion of remaining litter mass, main chemistry, and soil biotic and abiotic factors were assessed during decomposition, and then, we made an in‐depth analysis of the changes in the leaf litter decomposition. According to our results, leaf litter decomposition rate was significantly higher in the PK than that in the SF. Overall, the remaining percent mass of leaf litter''s main chemical quality in SF was higher than in PK, indicating that leaf litter chemical turnover in PK was relatively faster. PK had a significantly higher amount of total phospholipid fatty acids (PLFAs) than SF during decomposition. Based on multivariate regression trees, the forest type influenced the soil habitat factors related to leaf litter decomposition more than decomposition time. Structural equation modeling revealed that litter N was strongly and positively affecting litter decomposition, and the changes in actinomycetes PLFA biomass played a more important role among all the functional groups. Selected soil abiotic factors were indirectly driving litter decomposition through coupling with actinomycetes. This study provides evidence for the complex interactions between leaf litter substrate and soil physical–chemical properties in affecting litter decomposition via soil microorganisms.     

Structural and functional measures of leaf-associated invertebrates and fungi as predictors of stream eutrophication

Eutrophication is a major threat to freshwater ecosystems worldwide that affects aquatic biota and compromises ecosystem functioning. In this study, we assessed the potential use of leaf decomposition and associated decomposer communities to predict stream eutrophication. Because leaf quality is expected to affect leaf decomposition, we used five leaf species, differing in their initial nitrogen concentration. Leaves of alder, chestnut, plane, oak and eucalyptus were placed in coarse-mesh bags and immersed in six streams along an eutrophication gradient to assess leaf decomposition and the structure of associated decomposer communities. A hump-shaped relationship was established between leaf decomposition and the eutrophication gradient for all leaf species, except for eucalyptus. Invertebrate biomass and density as well as fungal biomass and sporulation were lowest at the extremes of the gradient. Leaf-associated invertebrate and fungal assemblages were mainly structured by stream eutrophication. The percentage of shredders on leaves decreased, whereas the percentage of oligochaeta increased along the eutrophication gradient. The Iberian Biological Monitoring Working Party Index (IBMWP) applied to benthic invertebrates increased from oligotrophic to moderately eutrophic streams and then dropped sharply at highly and hypertrophic streams. Overall, leaf decomposition was a valuable tool to assess changes in stream water quality, and it allowed the discrimination of sites classified by the IBMWP within class I and class IV. Moreover, decomposition of most leaf species responded in a similar way to eutrophication when decomposition was normalized by the quality of leaves.     

Anaerobic transformation of 1,1,1-trichloroethane by municipal digester sludge

Anaerobic transformations of 1,1,1-trichloroethane (TCA), 1,1-dichloroethane (DCA), and chloroethane (CA) were studied with sludge from a lab-scale, municipal wastewater sludge digester. TCA was biologically transformed to DCA and CA and further to ethane by reductive dechlorination. TCA was also converted to acetic acid and 1,1-dichloroethene (11DCE) by cell-free extract. 11DCE was further biologically converted to ethene. This pathway was confirmed by transformation tests of TCA, DCA and CA, by tests with cell-free extract, and by chloride release during TCA degradation. With cell-free extract, acetic acid accounted for approximately 90% of the TCA transformed; tests with live cells indicate that the fraction of TCA transformed by this pathway decreased with lower biomass. The dechlorination of DCA to CA and CA to ethane was not stoichiometric. A high rate of TCA removal was observed under the experimental conditions. The results indicate that removal of TCA in anaerobic digestion should be complete, but DCA and CA could persist in a normally operating digester.     

Cholesterol metabolism in liver at different levels of motor activity

Biosynthesis and decomposition of cholesterol were studied in the rabbit liver under a dosed physical loading. Judging from the intensity of [2-14C]acetate incorporation, the cholesterol biosynthesis intensity rises during loading and decreases under conditions of limited mobility. The cholesterol content in the liver tissue of trained animals is unchanged and in animals, who were under conditions of limited mobility, it rises as compared with the control. Using [7-3N1] cholesterol as a substrate, it established that the activity of cholesterol-7 alpha-hydroxylase, as compared to the initial level, is increased under training and decreased under conditions of low mobility. The enzymic activity was determined by the amount of released 3H2O.     

Organic chemical and nutrient dynamics in decomposing beech leaf litter in relation to fungal ingrowth and succession during 3-year decomposition processes in a cool temperate deciduous forest in Japan

Decomposition processes of beech leaf litter were studied over a 3-year period in a cool temperate deciduous forest in Japan. Organic chemical and nutrient dynamics, fungal biomass and succession were followed on upper (Moder) and lower (Mull) of a forest slope. Litter decomposition rates were similar between the sites. Nutrient dynamics of the decomposing litter was categorized into two types: weight changes in nitrogen and phosphorus showed two phases, the immobilization (0–21 months) and the mobilization phase (21–35 months), while those in potassium, calcium and magnesium showed only the mobilization phase. The rate of loss of organic chemical constituents was lignin < holocellulose < soluble carbohydrate < polyphenol in order. The changes in lignocellulose index (LCI), the ratio of holocellulose in lignin and holocellulose, were significantly correlated to the changes in nitrogen and phosphorus concentrations during the decomposition. During the immobilization phase, increase in total fungal biomass contributed to the immobilization of nitrogen and phosphorus. The percentage of clamp-bearing fungal biomass (biomass of the Basidiomycota) to total fungal biomass increased as the decomposition proceeded and was significantly correlated to LCI. Two species in the xylariaceous Ascomycota were dominantly isolated by the surface sterilization method from decomposing litter collected in the 11th month. The organic chemical, nitrogen and phosphorus dynamics during the decomposition were suggested to be related to the ingrowth, substrate utilization and succession of the Xylariaceae and the Basidiomycota. Twenty-one species in the other Ascomycota and the Zygomycota isolated by the washing method were classified into three groups based on their occurrence patterns: primary saprophytes, litter inhabitants and secondary sugar fungi. These species showed different responses to LCI and soluble carbohydrate concentration of the litter between the groups.     

Inhibition of sterol synthesis by citrinin in a cell-free system from rat liver and yeast.

Citrinin, a fungal metabolite known as an antibiotic, strongly inhibited the labeled acetate incorporation into nonsaponifiable lipids by a cell-free system from rat liver but not the labeled mevalonate incorporation. Of the enzymes involved in cholesterol synthesis, two enzymes, acetoacetyl-CoA thiolase (EC 2.3.1.9) and 3-hydroxy-3-methylglutaryl-CoA reductase (EC 1.1.1.34), were specifically inhibited by the antibiotic. The concentration required for 50% inhibition was 0.2 mM for the former enzyme and 0.5 mM for the latter. Essentially the same results were obtained with a cell-free system from yeast although higher concentrations of the antibiotic were required for inhibition.     

Microbial Biomass,Respiration, and Decomposition of Hura crepitans L. (Euphorbiaceae) Leaves in a Tropical Stream1

The processing of leaves in temperate streams has been the subject of numerous studies but equivalent tropical ecosystems have received little attention. We investigated leaf breakdown of a tropical tree species (Hura crepitans, Euphorbiaceae), in a tropical stream using leaf bags (0.5 mm mesh) over a period of 24 days. We followed the loss of mass and the changes in adenosine triphosphate (ATP) concentrations and respiration rates associated with the decomposing leaves. The breakdown rate was fast (k=?0.0672/d, k_d=?0.0031/degree‐day), with 81 percent loss of the initial mass within 24 days. This high rate was probably related to the stable and high water temperature (22°C) favoring strong biological activity. Respiration rates increased until day 16 (1.1 mg O₂/h/g AFDM), but maximum ATP concentrations were attained at day 9 (725 nmol ATP/g AFDM) when leaf mass remaining was 52 percent. To determine the relative importance of fungi and bacteria during leaf decomposition, ATP concentrations, and respiration rates were determined in samples treated with antibiotics, after incubation in the stream. The results of the samples treated with the antifungal or the bacterial antibiotic suggest a higher contribution of the fungal community for total microbial biomass and a higher contribution of the bacterial community for microbial respiration rates, especially during the later stages of leaf decomposition. However, these results should be analyzed with caution since both antibacterial and antifungal agents did not totally eliminate microbial activity and biomass.     

高山森林凋落物分解过程中的微生物生物量动态

凋落物分解过程中的微生物生物量动态对于深入了解森林凋落物分解机理具有重要意义。为了解高山森林典型树种凋落物分解过程中的微生物生物量特征,采用凋落物分解袋法,研究了土壤冻结期(3月)、融冻期(4月-5月)、生长季节(5-10月)和冻结初期(11月)红桦(Betula albosinensi)、岷江冷杉(Abies faxoniana)和粗枝云杉(Picea asperata)凋落物分解过程的微生物生物量C(MBC)、微生物生物量N(MBN)和微生物生物量P(MBP)动态。四个关键时期,凋落物的MBC、MBN以生长季节最高,但非生长季节的三个关键时期也检测出较高的MBC、MBN。在融冻期结束后,三类凋落物分解过程中MBC和MBN均出现爆发性增长。然而,MBP在生长季节中期(8月)、完全冻结期(3月)和冻结初期(11月)均相对较低,但在融冻期和生长季节后期(9月)相对较高。另外,红桦凋落物的MBC、MBN和MBP含量均高于岷江冷杉和粗枝云杉凋落物(除4月粗枝云杉凋落物MBP异常升高外)。这些结果为更加清晰地认识高寒森林凋落物分解过程及机理,以及进一步理解陆地生态系统结构和功能提供了一定基础数据。     

高山森林三种细根分解初期微生物生物量动态

为了解川西高山森林凋落物分解过程的微生物生物量特征,采用凋落物分解袋法,测定了粗枝云杉(Picea asperata)、岷江冷杉(Abies faxoniana)和红桦(Betula albosinensi)细根分解几个关键时期微生物生物量碳(MBC)、氮(MBN)和磷(MBP)的动态特征。3个树种细根分解过程中的MBC均表现为在土壤深冻期下降至全年最低点后缓慢上升,至土壤融冻中期再次下降,到生长季节增长的趋势。然而,粗枝云杉与岷江冷杉细根分解过程中的MBC最大值出现在生长季节末期,红桦细根分解过程中的MBC最大值出现在土壤冻结初期。3个树种细根分解过程中的MBN表现出相似的动态规律:土壤深冻期急剧下降至全年最低,随后在冻融季节无显著变化,生长季节明显增加,到生长季节末期达到全年最大值。另外,粗枝云杉和岷江冷杉细根分解过程中MBP均随着分解的进行呈现增加趋势,而红桦细根分解过程中的MBP在土壤融冻末期出现最大值,在生长季节中期出现另一峰值,生长季节末期明显下降。这些结果表明冬季细根分解过程中仍存在一定的土壤微生物,但受到细根质量、温度及其驱动的环境因子的深刻影响。     

Cell-free transfer of sterols from dictyosome-like structures to plasma membrane vesicles of guinea pig testes

Summary Transfer of radiolabeled lipids from dictyosome-like structures (DLS) from testis tubules of the guinea pig as donor to unlabeled plasma membrane from testis tubules immobilized on nitrocellulose as acceptor was studied in a completely cell-free system. As a general label for lipids of the donor DLS, isolated testis tubules were incubated with [¹⁴C]acetate. Time- and temperature-dependent transfer of [¹⁴C]acetate labeled constituents was observed in the cellfree system. However, despite the fact that phospholipids and other constituents were highly labeled in the donor fraction, primarily radioactive sterols were transferred to the plasma membrane acceptor vesicles. Transfer at 37°C represented 0.4 to 0.7% of the total radiolabeled cholesterol at 37°C but little or no transfer occurred at 4°C. The sterols transferred exhibited Chromatographic mobilities corresponding to those of cholesterol and lanosterol. Similar results were obtained with [¹⁴C]mevalonic acid. In subsequent experiments, cholesterol transfer from DLS to plasma membrane was demonstrated by incubation of DLS with [³H]squalene which was converted into sterol or with [¹⁴C]cholesterol. Transfer of sterols required ATP, but not cytosol, and was both time- and temperature-dependent. DLS were more effective than either endoplasmic reticulum or plasma membrane as the donor fraction. The results from the cell-free analysis suggest a possible functional role of the DLS in sterol biogenesis and transfer to the plasma membrane during spermatid development.Abbreviations DLS dictyosome-like structure(s) - PBS phosphatebuffered saline - HEPES 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid - BSA bovine serum albumin     

A fluorescent sterol probe study of cholesterol/phospholipid membranes

The behavior of dehydroergosterol in -α-dimyristoylphosphatidylcholine (DMPC) unsonicated multilamellar liposomes was characterized by absorption spectroscopy and fluorescence measurements. Dehydroergosterol exhibited a lowered absorption coefficient in multilamellar liposomes whiel the steady-state fluorescence anisotropy of dehydroergosterol in these membranes decreased significantly with increasing dehydroergosterol concentration, suggesting membrane sterol-sterol interactions. The comparative steady-state anisotropy of 0.9 mole percent dehydroergosterol in multilamellar liposomes was lower than in small unilamellar vesicles suggesting different sterol environments for dehydroergosterol. Dehydroergosterol fluorescence lifetime was relatively independent of membrane sterol content and yielded similar values in sonicated and unsonicated model membranes. In multilamellar liposomes containing 5 mole percent cholesterol, the gel-to-liqui crystalline phase transition of DMPC detected by 0.9 mole percent dehydroergosterol was significantly broadened when compared to the phase transition detected by dehydroergosterol in the absence of membrane cholesterol (Smutzer, G. et al. (1986) Biochim. Biophys. Acta 862, 361–371). In multilamellar liposomes containing 10 mole percent cholesterol, the major fluorescence lifetime of dehydroergosterol did not detect the gel-to-liquid crystalline phase transition of DMPC. Time-correlated fluorescence anisotropy decays of dehydroergosterol in DMPC multilamellar liposomes in the absence and presence of 5 mole percent cholesterol exhibited a single rotational correlation time near one nanosecond that was relatively independent of temperature and low concentrations of membrane cholesterol. The limiting anisotropy of 0.9 mole percent dehydroergosterol decreased above the gel-to-liquid crystalline phase transition in membranes without cholesterol and was not significantly affected by the phase transition in membranes containing 5 mole percent cholesterol. These results suggested hindered rotational diffusion of dehydroergosterol in multilamellar liposomes. Lifetime and time-correlated fluorescence measurements of 0.9 mole percent dehydroergosterol in multilamellar liposomes further suggested this fluorophore was detecting physical properties of the bulk membrane phospholipids in membranes devoid of cholesterol and was detecting sterol-rich regions in membranes of low sterol concentration.     

模拟氮沉降对华西雨屏区天然常绿阔叶林凋落叶分解过程微生物生物量的影响

为进一步深化氮沉降对凋落物分解影响的研究,2016年3月—2017年3月,在华西雨屏区天然常绿阔叶林内,用凋落叶分解袋法研究了模拟氮沉降对凋落叶分解过程中微生物生物量碳(MBC)、微生物生物量氮(MBN)和微生物生物量磷(MBP)的影响。实验设置了对照(0 g N m~(-2)a~(-1))、低氮(5 g N m~(-2)a~(-1))、中氮(15 g Nm~(-2)a~(-1))和高氮沉降(30 g N m~(-2)a~(-1)) 4个处理。结果表明:低氮和中氮处理显著增加了凋落叶分解过程中的MBC和MBN,以低氮处理增加幅度最高;低氮和中氮处理对凋落叶分解过程中的MBP影响不显著;高氮处理显著降低了分解过程中的MBC、MBN和MBP。随模拟氮沉降量的递增,凋落叶分解过程中微生物生物量碳氮比逐渐减少,微生物生物量碳磷比呈现先增加后下降的趋势。研究结果说明,氮沉降影响了华西雨屏区天然常绿阔叶林凋落物分解过程中微生物生物量,进而改变了凋落物的分解过程。