基于氮平衡的种养结合沼液资源化利用研究

研究沼液经济性消纳对提高沼液资源利用率、维持沼气工程稳定运行、缓解养殖带来的环境压力、提升农业废弃物资源化利用效率和经济效益,推动种养循环一体化发展具有重要意义。为此,结合农业废弃物资源化利用方式,基于盈亏平衡分析和秸秆资源化利用模型,以沼液经济性消纳半径为核心,探究了沼气工程投资金额和处理规模之间的关系,提出了沼液经济性消纳模型。研究结果表明:沼气工程投资金额与处理规模之间关系性较强(R 2=0.9842);沼液经济性消纳半径R bsa选取的主要影响因素是运输费用和沼液中N含量,不同消纳区域农业生产系数的差异性也会对经济性消纳半径的评估产生影响。利用模型对江西新余南英垦殖场规模化沼气发电工程(SBPGP)沼液经济性消纳半径进行估算,在保证氮平衡基础下,其沼液最小消纳范围在31.94 km 2左右。沼液经济性消纳模型的构建,为以沼气工程为纽带的种养一体化循环模式提供了技术参考,从经济性视角为循环农业发展提供了理论依据,从而实现经济效益与环境效益协调发展的目标。     

Classification of differentiating oocytes during ovarian cycle in the giant freshwater prawn, Macrobrachium rosenbergii de man

Based on the light microscopic observations of cells' sizes, chromatin patterns, amount of lipid droplets and yolk granules, the female germ cells could be classified into four different phases, which include 1) oogonia (Oog), 2) primary oocytes (pOc), 3) secondary oocytes (sOc), and 4) mature oocyte (mOc). Oog are small oval-shaped cells with irregular-shaped nuclei sizing 4–6 μm in diameter. They rest on the connective tissue germinal cord at the tip of each ovarian pouch (lobule). Oogonia increase their number through mitotic division, and the daughter cells move into ovarian pouch where they undergo first meiotic division to become primary oocytes, which have various steps of 1st meiotic prophase accumulating at the innermost zone of the ovarian pouch. The primary oocytes are small oval-shaped cells (8.5–10 μm in diameter) with large nuclei containing chromatin in various states of condensation that finally transform into chromatids. Their nuclei are surrounded by thin rim of faint blue-stained cytoplasm. The secondary oocytes derived from 2nd meiosis and comprise five steps: Oc1 and Oc2, classified as previtellogenic oocytes, Oc3 and Oc4, classified as vitellogenic oocytes, and mature oocyte (mOc) The zones of ovarian pouch are defined based on the accumulation of various steps of developing oocytes, namely, oogenic, previtellogenic, vitellogenic and mature zones, respectively. The ovarian cycle is divided into five stages based on the number and types of oocytes present in each stage. Stage 0 and I are spawn and spent stages. Stage II and III are proliferative and premature stages, while stage IV is mature stage. During ovarian stage I, each ovarian pouch contains primarily oogonia, primary oocytes, Oc1 and a few Oc2. In stage II, the pouch contains mainly Oc2 and Oc3, while in stage III the predominant cells are Oc4. Mature oocytes appear synchronously, in stage IV. The ovulating mature oocytes pass through the thin disrupted wall of ovarian pouch into subcapsular space, that leads into the oviduct situated on the ventro-lateral side of the ovarian lobe. At spawning, the ovarian pouches break down and only connective sheaths and hemolymph sinuses remain. The germinal cords and islets of oogonia remain in the central area of stage 0 ovary. The ovarian capsule, including the muscular layer, becomes attenuated as the ovary progresses from stage 0 to IV. The hemolymph vessels become highly convoluted in the central area of the ovary, and they branch radially into smaller hemolymph sinuses around each oogenic pouch.     

Cytotoxic Effects of Loperamide Hydrochloride on Canine Cancer Cells

Loperamide is a peripheral opiate agonist that can cause apoptosis and G2/M arrest in human cancer cell lines and may sensitize cells to chemotherapy. The objectives of this study were to investigate the effects of loperamide on viability, apoptosis and cell cycle kinetics in canine cancer cells and to establish whether the drug sensitizes cells to doxorubicin. Cell viability was assessed using Alamar Blue. Cell death and cell cycle were studied using flow cytometry with 7-Aminoactinomycin-D (7-AAD) and propidium iodide (PI), respectively. Loperamide decreased cell viability in a dose-dependent fashion and was most effective against canine osteosarcoma cells. In all cell lines, it induced a dose and time dependent apoptosis and resulted in accumulation in G0/G1. When co-incubated with doxorubicin, loperamide induced a synergistic cell kill in canine carcinoma cells. Investigation is warranted into the role of loperamide in the treatment of canine cancer.     

Cell-cycle-dependent Colonization of Mouse Spermatogonial Stem Cells After Transplantation into Seminiferous Tubules

Spermatogonial stem cells (SSCs) migrate to the niche upon introduction into the seminiferous tubules of the testis of infertile animals. However, only 5–10% of the transplanted cells colonize recipient testes. In this study, we analyzed the impact of cell cycle on spermatogonial transplantation. We used fluorescent ubiquitination-based cell cycle indicator transgenic mice to examine the influence of cell cycle on SSC activity of mouse germline stem (GS) cells, a population of cultured spermatogonia enriched for SSCs. GS cells in the G1 phase are more efficient than those in the S/G2-M phase in colonizing the seminiferous tubules of adult mice. Cells in the G1 phase not only showed higher expression levels of GFRA1, a component of the GDNF self-renewal factor receptor, but also adhered more efficiently to laminin-coated plates. Furthermore, this cell cycle-dependency was not observed when cells were transplanted into immature pup recipients, which do not have the blood-testis barrier (BTB) between Sertoli cells, suggesting that cells in the G1 phase may passage through the BTB more readily than cells in the S/G2-M phase. Thus cell cycle status is an important factor in regulating SSC migration to the niche.     

3-甲基喹噁啉-2-羧酸的制备及其细胞毒性

通过贝鲁特反应和还原、脱羧反应在体外合成制备3-甲基喹噁啉-2-羧酸,并进行理化鉴定和抗菌活性测试;通过MTT方法研究3-甲基喹噁啉-2-羧酸对多种细胞的生长抑制作用,利用单细胞电泳研究其对细胞DNA的损伤作用,利用流式细胞术研究其对细胞周期的改变。结果显示,体外成功合成制备出3-甲基喹噁啉-2-羧酸,而3-甲基喹噁啉-2-羧酸几乎没有抗菌效果,对多种细胞的生长抑制作用较弱,在剂量检测范围内细胞抑制率不到30%,但在一定剂量下能导致细胞DNA损伤,主要表现为尾长和尾部DNA含量显著升高;并且3-甲基喹噁啉-2-羧酸也能改变Chang细胞的细胞周期,表现为S期阻滞。结果表明,3-甲基喹噁啉-2-羧酸具有一定的细胞毒性。     

产蛋循环过程中TRPV6、CaBP-D_（28K）和PMCA 1b在蛋壳腺的动态表达

将40羽220日龄ISA蛋鸡分为5组,于产蛋后0、2、4.5、8、16h断头处死,采集蛋壳腺组织,运用Real-time PCR和Western-blot方法检测产蛋循环过程中瞬时性受体单位香草精受体6（Transient receptor potential vanilloid receptor 6,TRPV6）、钙结合蛋白（calbindin D28K,CaBP-D28K）、质膜钙离子ATP酶1b（plasma membrane Ca2＋-ATPase 1b,PMCA 1b）mRNA和蛋白浓度的动态变化。结果显示,卵子未进入蛋壳腺前（产蛋后0~4.5h）,蛋壳腺内TRPV6、CaBP-D28K和PMCA 1bmRNA表达水平较低,随后表达量逐渐升高,在蛋壳钙化过程中达到最大值（产蛋后16h）,与0h相比,TRPV6和CaBP-D28KmRNA表达差异均极显著（P〈0.01）;另外,产蛋循环过程中,TRPV6、CaBP-D28K和PMCA 1b蛋白浓度变化与mRNA变化一致,产蛋后16h到达最大值,其中CaBP-D28K蛋白浓度与0h相比,差异显著（P〈0.05）。结果表明,产蛋循环可调控蛋壳腺内TRPV6、CaBP-D28K和PMCA 1b的表达,并提示钙离子跨细胞转运途径在钙离子进入蛋壳腺形成蛋壳过程中发挥重要作用。     

文蛤的性腺发育和生殖周期

应用组织学方法对浙南池塘养殖的文蛤的性腺发育、生殖周期和肥满度进行了周年变化的研究。结果表明,文蛤的性腺发育具有典型双壳贝类特征,性腺发育过程可分为增殖期、生长期、成熟期、排放期、休止期5个时期,并对上述5期的宏观肉眼观察特征及微观切片观察特征进行了描述。浙南池塘养殖的文蛤性腺发育过程以1年为1个周期,成熟排放期在5月下旬至8月中旬,水温21.8～31.0℃,繁殖盛期为6月下旬至7月下旬,水温为25.0～30.0℃。月平均肥满度最高为6月份的8.84%,最低为1月份的3.21%。论文还讨论了1龄文蛤性腺发育、文蛤性腺成熟和排放与积温的关系。     

养殖水环境中氮的循环与平衡

养殖水环境中，由于人为的介入和影响。使得水体中氮的各种形态转化和循环过程中发生了改变。笔者详细地介绍了养殖水环境中氮的各种存在形态，及其相互之间的转化和循环过程，同时总结了前人研究和提出的各种养殖水环境中氮的物质平衡模型，为优化水产养殖和保护水环境提供科学依据。     

The life cycle of Henneguya nuesslini Schuberg & Schroder, 1905 (Myxozoa) involves a triactinomyxon-type actinospore

The life cycle of the histozoic myxozoan parasite Henneguya nuesslini was investigated in two salmonid host species. Naive brown trout, Salmo trutta, and brook trout, Salvelinus fontinalis, were experimentally infected in two trials by triactinomyxon type actinospores from naturally infected Tubifex tubifex. In exposed common carp, Cyprinus carpio, no myxospore production was detected. The parasite formed cysts with mature myxospores in the connective tissue of the fish 102 days post-exposure. The morphology of both actinosporean and myxosporean stages was described by light microscopy and a 1417-bp fragment of the 18S rDNA gene was sequenced. Sequence analysis confirmed the absolute congruence of the two developmental stages and assisted in determining species identity. Host range, tissue specificity and myxospore measurements provided sufficiently distinctive features to confirm species validity and were thus crucial for identification. The triactinomyxon spores had 16 secondary germ cells, unique dimensions, a very opaque sporoplasm matrix and three conspicuously protruding, pyriform polar capsules. This is the first record of a Henneguya sp. life cycle with a triactinomyxon-type actinospore, which suggests a close relationship with the Myxobolus group and a polyphyletic origin of the genus Henneguya.     

RASSF1A expression mediated by lentivirus inhibits growth of small-cell lung cancer cell line H446

AIM：To explore the inhibitory effect of Ras-association domain family 1A (RASSF1A) on the small-cell lung cancer cell growth. METHODS：The lentiviral expression vector containing RASSF1A gene was constructed and used to infect the small-cell lung cell line H446. The growth curve and cell cycle were detected by MTT assay and flow cytometry. The mRNA and protein levels of cell cycle-associated proteins were determined by real-time PCR and Western blotting. RESULTS：We obtained the H446 cells in which RASSF1A was stably expressed (named RASSF1A-H446). Compared with normal cell group and negative cell group, RASSF1A inhibited the proliferation of H446 cells, and arrested H446 cells in G1 phase. The expression of p21 and p27 was significantly increased, and E2F1 was significantly decreased in RASSF1A-H446 cells. CONCLUSION：RASSF1A inhibits the H446 cell growth by increasing the expressions of p21 and p27, and decreasing the expression of E2F1.