A New Micropropagation Technology of Tilia amurensis:In VitroMicropropagation of Mature Zygotic Embryos and the Establishment of a PlantRegeneration System

Tilia amurensis is an economically valuable broadleaf tree species in Northeast China.The production of highqualityT.amurensis varieties at commercial scales has been greatly limited by the low germination rates.Thereis thus a pressing need to develop an organogenesis protocol for in vitro propagation of T.amurensis to alleviate ashortage of high-quality T.amurensis seedlings.Here,we established a rapid in vitro propagation system forT.amurensis from mature zygotic embryos and analyzed the effects of plant growth regulators and culture mediain different stages.We found that Woody plant medium(WPM)was the optimal primary culture medium formature zygotic embryos.The highest callus induction percentage(68.76%)and number of axillary buds induced(3.2)were obtained in WPM+0.89μmol/L 6-benzyladenine(6-BA)+0.46μmol/L kinetin(KT)+0.25μmol/Lindole-3-butryic acid(IBA)+1.44μmol/L gibberellin A_(3)(GA_(3)).The multiple shoot bud development achievedthe highest percentage(83.32%)in the Murashige and Skoog(MS)+2.22μmol/L 6-BA+0.25μmol/L IBA+1.44μmol/L GA_(3).The rooting percentage(96.70%)was highest in 1/2 MS medium+1.48μmol/L IBA.Thesurvival percentage of transplanting plantlets was 82.22%in soil:vermiculite:perlite(5:3:1).Our study is the firstto establish an effective organogenesis protocol for T.amurensis using mature zygotic embryos.

Production of marker-free transgenic plants from mature tissues of navel orange using a Cre/loxP site-recombination system

Genetic transformation with mature material as the explants could shorten the transgenic period and avoid seed dependence compared with genetic transformation using the epicotyl seedling stem segments as the receptor. Here, we constructed an Agrobacterium tumefaciensmediated transformation for generation of marker-free transgenic plants from navel orange(Citrus sinensis Osbeck) mature stems using a CreloxP recombination system. To efficiently recover the regenerated buds from mature tissues, five recovery methods were compared: in vitro micrografting of 0.1-0.5(1-2 weeks), > 0.5 cm(3-4 weeks) and > 1 cm long lignified bud and in vitro micrografting of explants with a bud and rooting regenerated bud. The data showed that in vitro micrografting of > 1 cm long regenerated bud with expanded leaves after one month of continuous culture for lignification was the optimal solution for plant recovery from mature tissues. Transgenic plants without selectable marker genes were created from navel orange(Citrus sinensis Osbeck) tissue using a transformation vector PLI-35SPR1aCB containing a Cre/loxP system recombination together with genes encoding the selectable marker isopentenyl transferase(IPT) and an anti-bacterial peptide(PR1aCB).Using IPT positive selection, the transformation efficiency determined by PCR was 0.9%, and in total, 20 transgenic plants were obtained.Southern blotting confirmed further their transgenicity. PCR and sequencing analysis demonstrated that both the Cre and IPT genes had been successfully removed from the transgenic plants(deletion efficiency 100%). Over all, using Cre/loxP system recombination together with the IPT positive selection, marker-free transgenic plants can be recovered efficiently from mature tissues of navel orange(Citrus sinensis Osbeck), which provides a potential method for production of transgenic plants from citrus mature tissue.

Lamellar Bodies Count (LBC) as a Predictor of Fetal Lung Maturity in Preterm Premature Rupture of Membranes Compared to Neonatal Assessment

Background: Respiratory distress syndrome (RDS) is a major cause of neonatal morbidity and mortality, affecting approximately 1% of all live births and 10% of all preterm infants. Lamellar bodies represent a storage form of pulmonary surfactant within Type II pneumocytes, secretion of which increases with advancing gestational age, thus enabling prediction of the degree of FLM. Preterm premature rupture of membranes (PPROM) complicates approximately 1/3 of all preterm births. Birth within 1 week is the most likely outcome for any patient with PPROM in the absence of adjunctive treatments. Respiratory distress has been reported to be the most common complication of preterm birth. Sepsis, intraventricular haemorrhage, and necrotizing enterocolitis also are associated with prematurity, but these are less common near to term. Objective: To assess the efficacy of the amniotic fluid lamellar body counting from a vaginal pool in predicting fetal lung maturity in women with preterm premature rupture of membranes. Methods: This study was conducted at Ain Shams University Maternity Hospital in the emergency ward from January 2019 to September 2019. It included 106 women with singleton pregnancies, gestational age from 28 - 36 weeks with preterm premature rupture of membranes. This study is designed to assess the efficacy of the amniotic fluid lamellar body counting (LBC) from a vaginal pool in predicting fetal lung maturity in women with preterm premature rupture of membranes. Results: The current study revealed a highly significant increase in the lamellar body count in cases giving birth to neonates without RDS compared to that cases giving birth to neonates with RDS. Also, no statistically significant difference between LBC and age, parity and number of previous miscarriages in the mother was found. Gestational age at delivery was significantly lower among cases with respiratory distress. Steroid administration was significantly less frequent among cases with respiratory distress. However, lamellar bodies had high diagnostic performance in the prediction of respiratory distress. Conclusion: Lamellar body count (LBC) is an effective, safe, easy, and cost-effective method to assess fetal lung maturity (FLM). It does not need a highly equipped laboratory or specially trained personnel, it just needs the conventional blood count analyzer. Measurement of LBC is now replacing the conventional Lecithin/Sphyngomyelin L/S ratio. LBC cut-off value of ≤42.5 × 103/μL can be used safely to decide fetal lung maturity with sensitivity of 95.7% and specificity of 97.6%.

Debugging Strategies for Parents' Educational Anxiety in the Transition from Kindergarten to Primary School--Based on the Theory of Mature Forces

Due to the rapid growth of the economy and society,the burden of social security has been increasingly escalating,resulting in a continuous rise in parenting stress among certain groups of parents.On one hand,these parents aspire for their young children to grow through play;on the other hand,they are acutely aware that early childhood education constitutes the foundational stage of basic education and serves as the commencement phase of lifelong learning.Thus,they recognize the importance of emphasizing early childhood education.Therefore,preschool education has been given important significance.Parents place particular emphasis on education during this stage.They actively leverage their educational,financial,and social advantages to create favorable educational resources and opportunities for their young children.However,to a certain extent,this exacerbates educational anxiety among a segment of parents.Despite the implementation of various governmental measures,it appears that these initiatives have not fundamentally addressed parental educational anxiety at its root.This study employs interview techniques and literature review methodologies to gain a profound understanding of parental educational anxiety during the transition from early childhood to primary education.By integrating Gesell's Maturation Theory,this research aims to scientifically explicate the developmental patterns of children.Consequently,it endeavors to enable non-professional parents to adopt a"professional"perspective in their approach towards early childhood education.

Transcript Profiling Reveals Abscisic Acid,Salicylic Acid and Jasmonic-Isoleucine Pathways Involved in High Regenerative Capacities of Immature Embryos Compared with Mature Seeds in japonica Rice

Induced pluripotent cell mass plays a role in genetic transformation mediated by Agrobacterium. Mature seeds are more recalcitrant to the induction of suitable calli than immature embryos in rice, but the exact molecular mechanisms involved remain elusive. In this study, the morphological structure of calli induced from mature seeds and immature embryos were observed under a scanning electron microscope using a paraffin embedded technique. Meanwhile, a total of 2 173 up- and down-regulated genes were identified in calli induced from mature seeds and immature embryos by RNA-seq technique and furtherly confirmed by quantitative real-time PCR. The results revealed the remarkable morphological differences in calli induced from mature seeds and immature embryos, and plant hormone signal transduction and hormone biosynthesis pathways, such as abscisic acid, salicylic acid and jasmonic-isoleucine, were found to play roles in somatic embryogenesis. This study provided comprehensive gene expression sets for mature seeds and immature embryos that were served as an important platform resource for further functional studies in plant embryogenesis.

Comparative proteomic analysis of mature and immature oocytes in domestic cats

Objective:To evaluate changes of feline(Felis catus)oocytes proteins during in vitro maturation by using the proteomic approach.Methods:Immature oocytes(germinal vesicle)isolated from female cats were cultured and collected at 0 h and 24 h.After collection,oocytes were investigated into immature(germinal vesicle)and mature(metaphaseⅡ)stages.The qualitative profiles of the proteins at the immature and mature stages were determined by one-dimensional electrophoresis and liquid chromatography-mass spectrometry.Results:Our data revealed that following 24 h in vitro maturation the maturation rate(metaphaseⅡstage)was 58.7%.Eighty-one of the 260 proteins analyzed were differentially expressed between the germinal vesicle stage and the metaphaseⅡ-arrest stage.Proteomic analysis of germinal vesicle and metaphaseⅡoocytes showed abundant expression of proteins involved in transportation(10%),indicating that this was a major characteristic of germinal vesicle oocytes.Similarly,analysis of the proteome of metaphaseⅡoocytes indicated that cell cycle proteins were overexpressed.Interestingly,proteins involved in DNA repair and apoptosis were only expressed in germinal vesicle oocytes and proteins involved in fertilization were only expressed in metaphaseⅡoocytes.Conclusions:The overexpression of certain proteins in germinal vesicle and metaphaseⅡis necessary for oocyte development and maturation.Our findings provide a valuable resource for further investigations into protein expression in oocytes at different developmental stages.

Establishment of a Highly Efficient Regeneration System for the Mature Embryo Culture of Wheat

Establishment of a highly efficient regeneration system for the mature embryo of wheat will provide a convenient tool for wheat tissue culture and transformation, thereby facilitating the transformation of foreign genes into wheat. By using the mature embryos derived from 20 different wheat lines including Shi 4185, Yumai 66, Lunxuan 987, CB037, Yangmai 6, Xinchun 9, Bobwhite, Han 6172, Zheng 9023, Jimai 20, Ningchun 4, and Jing 411, the effects of some factors including inoculation methods, initiating culture media, organic additives, antioxidants, and auxins on the regeneration from the explants were evaluated. The results indicated that the scraping embryo culture was better than the whole embryo culture, the Aa medium was better than the SD2 medium and dicamba was better than 2,4-D in increasing the regeneration frequency. An Adi medium was established in this study by adding silver nitrate, cysteine, ascorbic acid, dicamba, glutamine into the Aa medium at the concentration of 4,40, 100, 2, and 5 mg L^-1, respectively. By using the Adi medium and the scraping technique, the regeneration frequencies of the mature embryos of CB037, Lunxuan 987, Hart 6172, Yangmai 6, Bobwhite, Zheng 9023, Shi 4 185, and Jimai 20 became 85.6, 60,1, 46.0, 42.1,42.0, 34.0, 33.0, and 32.0%, respectively, which were about 5-8 times higher than that obtained from the conventional culture mediums and techniques. This novel regeneration system could be helpful in wheat transformation.

Organic matter and concentrated nitrogen removal by shortcut nitrification and denitrification from mature municipal landfill leachate

An UASB＋Anoxic/Oxic （A/O） system was introduced to treat a mature landfill leachate with low carbon-to-nitrogen ratio and high ammonia concentration. To make the best use of the biodegradable COD in the leaehate, the denitrifieation of NOx^--N in the reeireulation effluent from the elarifier was carried out in the UASB. The results showed that most biodegradable organic matters were removed by the denitrifieation in the UASB. The NH4^＋-N loading rate （ALR） of A/O reactor and operational temperature was 0.28- 0.60 kg NH4^＋-N/（m^3-d） and 17-29℃ during experimental period, respectively. The short-cut nitrification with nitrite accumulation efficiency of 90%-99% was stabilized during the whole experiment. The NH4^＋-N removal efficiency varied between 90% and 100%. When ALR was less than 0.45 kg NH4^＋-N/（m^3.d）, the NH4^＋-N removal efficiency was more than 98%. With the influent NH4^＋-N of 1200-1800 mg/L, the effluent NH4^＋-N was less than 15 mg/L. The shortcut nitrification and denitrifieation can save 40% carbon source, with a highly efficient denitrifieation taking place in the UASB. When the ratio of the feed COD to feed NH4^＋-N was only 2-3, the total inorganic nitrogen （TIN） removal efficiency attained 67%-80%. Besides, the sludge samples from A/O reactor were analyzed using FISH. The FISH analysis revealed that ammonia oxidation bacteria （AOB） accounted for 4% of the total eubaeterial population, whereas nitrite oxidation bacteria （NOB） accounted only for 0.2% of the total eubaeterial population.

Study on Plant Regeneration of Wheat Mature Embryos Under Endosperm-Supported Culture

To reveal the suitability of using mature embryos as an explant source in wheat tissue culture, mature embryos from eight common wheat cultivars （Triticum aestivum L. cv.） were cultured with or without endosperm to test their efficiency of callus induction and plant regeneration. When embryos were cultured together with endosperm （endosperm-supported culture, ES）, the percentage of callus induction was significantly lower than that when embryos were cultured in the absence of endosperm （non-endosperm-supported culture, NES）. This pattern was evident in most genotypes, regardless of whether 2 or 8 mg L^-1 2,4-D was added in the NES culture. However, in ES culture, more induced calli were differentiated into distinct green spots and they further developed into plantlets. Thus, more plants were regenerated in ES culture than in the NES treatment. Most of the eight tested genotypes showed a significant difference in callus induction rate and plantlet regeneration in both ES and NES cultures. In addition, the enzymatic activity of oxalate oxidase in the callus of ES culture condition was obviously higher than that in the callus of NES culture condition, suggesting that the activity of oxalate oxidase may be a parameter for selection of calli with potential for plantlet regeneration. These results indicate that wheat mature embryos are valuable explants for highly efficient callus induction and plant regeneration, if proper treatment and medium are used.

Species diversity,population structure and regeneration of woody species in fallows and mature stands of tropical woodlands of southeast Angola

This study in the dry tropical woodlands of SE Angola in Cuando-Cubango Province assessed the diversity and composition of woody species in fallows compared to those in mature woodlands. We assessed the population structure of the most harvested woody species by calculating size class distribution and evaluated their regeneration potential based on the density of saplings. The vegetation was surveyed in 20 plots of 20 × 50 m(1000 m2). In each plot, we measured the diameter at breast height(DBH) of all woody species with DBH ≥ 5 cm. The saplings were counted, identified and recorded; 718 individuals, corresponding to 34 species in 32 genera and 15 families were recorded. The size class distribution of target woody species showed three different patterns in fallows and mature woodlands. In general,most of the smaller diameter classes had more individuals than the larger ones did, showing that the regeneration may take place. However, in some diameter classes, the absence of larger stems indicated selective logging of larger trees. Few saplings were recorded in the fallows or mature woodlands; fire frequency and intensity is probably the main obstacle for seed germination and seedling survival rates in the studied area.

Dicamba and Sugar Effects on Callus Induction and Plant Regeneration from Mature Embryo Culture of Wheat

To establish a highly efficient plant regeneration system for wheat genetic transformation, the effects of three different concentrations of dicamba and two different sugar types on callus induction and plant regeneration from mature embryo cultures were evaluated. Callus induction and plant regeneration were obtained from mature embryos of two commercial cultivars Zhoumai 18 and Yumai 34 （Triticum aestivum L.） cultured on L3 basal medium. The results showed that the efficiency of mature embryo culture was significantly influenced by the genotypes, sugar types and dicamba concentrations. 4 mg L^-1 dicamba proved the best effective for inducing embryogenic callus and also gave the highest proportion of plants regenerated across the two cultivars. Substitution of maltose by sucrose significantly improved the plant regeneration efficiency in both cultivars. There was a significant interaction between genotype-by-sugar types, and sugar types-bydicamba concentrations. Overall, Zhoumai 18 gave the highest frequency of plant regeneration （82.65%） when dicamba concentration was 4.0 mg L^-1 and with sucrose in initial callus induction. These results will facilitate genetic transformation work with elite wheat.

The Comparison in Tissue Culture Ability of Mature Embryo in Different Cultivars of Rice

In order to study the regeneration technology of mature embryos in different rice varieties,nine japonica,nine indica and eleven hybrid rice varieties of two line or three line or superiority combinations were selected as explants to study the callus induction,differentiation and regeneration rates on different media.The higher callus induction （61.7-89.2%） was observed in japonica rice,when cytokinin was added at lower concentration （0.3 mg L-1 6-BA） in M8 basal medium,supplemented with 30 g L-1 sucrose,8 g L-1 agar and 2 mg L-1 2,4-D.Further,the addition of two cytokinins （2 mg L-1 6-BA,0.5 mg L-1 KT） and 1 mg L-1 NAA in the M8 basal supplemented medium resulted in 9.1-100% of the callus induction in indica rice.The percent callus induction in hybrid rice varieties was 40-86.3% when addition of 1 mg L-1 6-BA and 1 mg L-1 KT was added,and the cytokinins was required by the japonica and indica rice varieties in the M8 basal supplemented medium.It was observed that when the 0.5 mg L-1 2,4-D and 1 mg L-1 6-BA were added in japonica rice,and 0.2 mg L-1 2,4-D and 0.5 mg L-1 6-BA were added in indica and hybrid rice in the MS different media,the regeneration rates were 9.2-59.5%,3.6-87.5% and 17.2-43.2% for japonica,indica and hybrid rice,respectively.Thus,the regeneration technology with higher output is established in the mature embryos of similar rice varieties.

Varietal Difference in Leaf Nitrogen Content and Leaf Area and Their Effects to Ripening Rate During Mature Period of japonica Rice

Employing the pot experiment of the complete random block design with 6 replications,four varieties of japonica rice (Fujisaka 5,Honenwase,Akitakomachi and Taichung 65) were used to study the varietal differences in leaf nitrogen content(LNC) and leaf area during mature period,their relation and effects to the ripening rate.The results showed that(1) thee were varietal differences in LNC at the heading stage and the LNC decrease rate during the matue period,the high LNC at the heading stage was related to the rapid LNC decrease.(2) There were two phases of the leaf area changing process during the mature period,first was the stable,and second was the decreased phase.There was varietal difference in the critical time of phase 1 and phase 2.The hign leaf area in the phase 1 was in relation to the rapid leaf area decrease in the phase 2.It was not found that there was relation between the leaf quality and quantity.(3)It wa unfavorable to the ripening rate for the high leaf area at the heading stage and the rapid decrease of the leaf area during the mature period.(4)It was put forward that the super high yield rice variety should possess the not very high leaf area and high LNC at the heading stage,slow senescence in the leaf area during the mature period.

Evolution and organic geochemical significance of bicyclic sesquiterpanes in pyrolysis simulation experiments on immature organic-rich mudstone

Sesquiterpanes are ubiquitous components of crude oils and ancient sediments.Liquid saturated hydrocarbons from simulated pyrolysis experiments on immature organic-rich mudstone collected from the Lower Cretaceous Hesigewula Sag were analyzed by gas chromatography-mass spectrometry(GC-MS).C14 bicyclic sesquiterpanes,namely,8β(H)-drimane,8β(H)-homodrimane,and 8 a(H)-homodrimane were detected and identified on basis of their diagnostic fragment ions(m/z123,179,193,and 207),and previously published mass spectra data,and these bicyclic sesquiterpanes presented relatively regular characteristics in their thermal evolution.The ratios 8β(H)-drimane/8β(H)-homodrimane,8β(H)-homodrimane/8 a(H)-homodrimane,and 8β(H)-drimane/8 a(H)-homodrimane all show a clear upward trend with increasing temperature below the temperature turning point.Thus,all these ratios can be used as evolution indexes of source rocks in the immature-lowmaturity stage.However,the last two ratios may be more suitable than the first ratio as valid parameters for measuring the extent of thermal evolution of organic matter in the immature-low-maturity stage because their change amplitude with increasing temperature is more obvious.

Expression of nerve growth factor precursor, mature nerve growth factor and their receptors during cerebral ischemia-reperfusion injury

We investigated nerve growth factor precursor （proNGF） and mature NGF expression in ischemic and non-ischemic cortices after cerebral ischemia-reperfusion injury. In both ischemic and non-ischemic cortices, proNGF was found to be present in the extracellular space and cytoplasm. In addition, mature NGF was expressed in extracellular space, but with a very low signal. In ischemic cortex only, proNGF was significantly decreased, reaching a minimal level at 1 day. Mature NGF was increased at 4 hours, then reached a minimal level at 3 days. The p75 neurotrophin receptor （p75NTR） was significantly decreased after ischemia, and increased at 3 days after ischemia. These results confirmed that proNGF was the predominant form of NGF during the pathological process of cerebral ischemia-repeffusion injury. In addition, our findings suggest that ischemic injury may influence the conversion of proNGF to mature NGF, and that proNGF/p75NTR may be involved in reperfusion injury.

Establishment and Optimization of the Regeneration System of Mature Embryos of Maize (Zea mays L.)

A reliable system was developed for regeneration from mature embryos derived from callus of four maize inbred lines （Liao 7980, Dan 9818, Dan 340, and Dan 5026）. The protocol was mainly based on a series of experiments involving the composition of culture medium. We found that 9 pM 2,4-dichlorophenoxyacetic acid in MS medium was optimum for the induction of callus. The induction frequency of primary calli was over 85% for four inbred lines tested. The addition of L- proline （12 mM） in subculture medium significantly promoted the formation of embryogenic callus but it did not significantly enhance growth rate of callus. Efficient shoot regeneration was obtained on regeneration medium containing 2.22 μM 6- benzylaminopurine in combinations with 4.64 μM Kinetin. Regenerated shoots were rooted on half-strength MS medium containing 2.85 μM indole-3-butyric acid. This plant regeneration system provides a foundation for genetic transformation of maize.

Genetic Analysis and QTL Mapping of Mature Seed Culturability in Indica Rice

Genetic segregation analysis for mature seed culturability was conducted using recombinant inbred lines derived from a cross of indica rice, Yangdao 6 and Pei＇ai 64s. Three indices of seed culturability, the frequency of callus induction, the frequency of brown callus and the increase of callus weight were investigated. A combined genetic map constructed with simple sequence repeat （SSR）, sequence tag site （STS）, cleaved amplified polymorphic sequences （CAPS） and single nucleotide polymorphism （SNP） markers covered a total distance of 1 732.5 cM, averaging approximately 12 cM between two neighboring loci. Three QTLs on chromosomes 7, 7 and 10 were detected for the frequency of callus induction; three QTLs on chromosomes 6, 7 and 9 were detected for the frequency of brown callus; and two QTLs on chromosomes 5 and 7 were detected for the increase of callus weight. Common QTLs mapped at the interval flanking RM5481 and RM6835 on chromosome 7 were identified to be involved in the frequency of callus induction and the frequency of brown callus, explaining 7.29% and 12.52% of phenotypic variation, respectively. A total of 14 epistatic effects were detected for the three indices of mature seed culturability. ~

Nutrient Accumulation and Distribution of Mature Pinus massoniana Plantation in Northwestern Guangxi

[Objectives]This study was conducted to reveal the characteristics of nutrient absorption and accumulation in Pinus massoniana plantations in Northwestern Guangxi.[Methods]Based on field investigation and indoor analysis,the contents,accumulation and annual net accumulation of five nutrient elements(N,P,K,Ca and Mg)in a mature P.massoniana plantation(26-year-old)in Nandan County,Guangxi Province were studied.[Results]The contents of nutrient elements in different organs of the mature P.massoniana plantation were the highest in the leaves,followed by the bark,branch and root,and the lowest in the stem.In general,among the contents of the five elements in different organs,N content was the highest,followed by K or Ca,and P and Mg were the lowest.The total accumulation of nutrient elements in the 26-year-old mature P.massoniana plantation in northwestern Guangxi was 1 384.05 kg/hm^2.Among the different structural levels of the stand,the tree layer had the highest accumulation of nutrient elements,which was 1 198.41 kg/hm^2,accounting for 86.59%of the total accumulation of nutrients in the plantation,and the accumulation of nutrients in other layers from the largest to the smallest was the litter layer(91.97 kg/hm^2),herb layer(49.86 kg/hm^2)and shrub layer(43.92 kg/hm^2),accounting for 3.17%,3.60%and 6.64%of the total nutrient accumulation of the plantation,respectively.The annual net accumulation of nutrient elements in the tree layer of the mature P.massoniana plantation was 46.09 kg/(hm^2·a),and the order of the annual net accumulation of different nutrient elements followed N>K>Ca>Mg>P;and the accumulation of 1 t of dry matter needed 6.37 kg of the five nutrients.[Conclusions]This study provides a scientific basis for the rational management of P.massoniana plantations,especially forest soil management.

THE INSEMINATION OF GOLDFISH(CARASSIUM AURATUS) OOCYTE MATURED IN VITRO

Full maturation of goldfish oocyte was induced in vitro by 17 a-hydroxy-20B-dihydroprogesterone. The oocyte maturation involves GV migration to the periphery of the oocyte and germinal vesicle breakdown (GVBD). In the experiment, incubation duration for GVBD varied in different broods of oocytes. Generally, if the duration for GVBD was shorter than 6 h, oocytes would have a better chance to survive after maturation and insemination. The maturation of nucleus (GV) and cytoplasm are not synchronous. Cytoplasm maturation occurs several hs after GVBD. Oocytes inseminated 8-9 h after GVBD have the highest fertilizing and hatching rate. Fertilized ova matured in vitro can develop to sexually mature adults capable of reproduction.

Influence of strong preformed particle gels on low permeable formations in mature reservoirs

In mature reservoirs,the success of preformed particle gel（PPG） treatment rests primarily on the ability of the PPG to reduce and/or plug the high permeable formations,but not damage the low permeable formations.Static test models（filtration test model and pressure test model）were used to determine the effect of PPG on low permeable formations.This work used a strong preformed particle gel,Daqing（DQ） gel made by a Chinese company.The particle gel sizes were ranged from 30 to 120 mesh for this work.PPGs are sized in a millimeter or micrometer,which can absorb over a hundred times their weight in liquids.The gel strength was approximately 6500 Pa for a completely swollen PPG with 1 %（weight percentage） NaCl solution（brine）.0.05 %,1 %,and 10 % NaCl solutions were used in experiments.Sandstone core permeability was measured before and after PPG treatments.The relationship between cumulative filtration volumes versus filtration times was determined.The results indicate that DQ gels of a particle size of 30–80 mesh did not damage the cores of a low permeability of 3–25 m D.The DQ gels of a smaller particle size ranging from 100 to 120 mesh damaged the core and a cake was formed on the core surface.The results also indicate that more damage occurred when a high load pressure（400 psi） was applied on the high permeability cores（290–310 m D）.The penetration of the particle gelsinto the low permeable formations can be decreased by the best selection of gel types,particle sizes,and brine concentrations.