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The effects and mechanism of GABAergic neurons, NOergic neurons, opioid peptide and cyclic adenosine monophosphate in the nucleus reticularis thalami on sleep-wakefulness cycle of rats and the effects and mechanism of the 5-HTergic nerve fibers project from the nucleus raphes dorsalis to RT on sleep-wakefulness cycle of rats were investigated with the methods of brain stereotaxic, nucleus spile, microinjection and polysomngraphy.1. The effects of GABAergic neurons in RT on sleep-wakefulness cycle of rats1.1 Microinjection of 3-mercaptopropionic acid (3-MP, a kind of glutamate decarboxylase inhibitor) into RT. On the day of microinjection, sleep only decreased a litter. On the second day, sleep marked decreased and wakefulness marked increased. On the third and fourth day, sleep and wakefulness stages resumed to normal.1.2 Microinjection of gamma-amino butyric acid (GABA 1.0μg) into RT enhanced sleep and reduced wakefulness compared with control; while microinjection of L-glutamate (L-Glu, 0.2μg) decreased sleep and increased wakefulness; microinjection of bicuculline (BIC, 1.0μg), a GABAA receptor antagonist, enhanced wakefulness and reduced sleep; microinjection of baclofen (BAC, 1.0μg), GABAB receptor agonist, had the same effects as GABA.2. The effects of NOergic neurons in RT on sleep-wakefulness cycle of rats2.1 Microinjection of L-arginine (L-Arg, 0.5μg) into RT decreased sleep compared with control, but there were on statistaical difference between L-Arg group and control; while microinjection of sodium nitroprusside (SNP, 0.2μg), a NO donor into RT, sleep marked decreased and wakefulness marked increased. Microinjection of nitric oxide synthase inhibitor, N-nitro-L-arginine (L-NNA, 2.0μg) into RT enhanced sleep and reduced wakefulness.2.2 After simultaneous microinjection of L-NNA (2.0μg) and SNP (0.2μg) into RT, SNP abolished the sleep-promoting effect of L-NNA compared with L-NNA group; after simultaneous microinjection of L-NNA (2.0μg) and L-Arg(0.5μg) into RT, we found that L-NNA could not blocked the wakefulness-promoting effect of L-Arg.3. The effects of opioid peptide in RT on sleep-wakefulness cycle of rats3.1 Microinjection of morphine sulfate (MOR, 1.0μg) into RT increased wakefulness and decreased sleep compared with control; while microinjection of naloxone hydrochloride (NAL, 1.0μg), the antagonist of opiate receptors, into RT, enhanced sleep and reduced wakefulness.3.2 After simultaneous microinjection of MOR (1.0μg) and NAL (1.0μg) into RT, the wakefulness-promoting effect of MOR and the sleep-promoting effect of NAL were not observed compared with control.4. The effects of cAMP in RT on sleep-wakefulness cycle of rats Microinjection of cAMP (1.0μg) into RT increased sleep and decreased wakefulness compared with control; microinjection of methylene blue (MB,1.0μg) into RT enhanced sleep and reduced wakefulness compared with control.5. The effects of the 5-HTergic nerve fibers project from DRN to RT on sleep-wakefulness cycle of rats5.1 When L-Glu (0.2μg) was microinjected into DRN and normal sodium (NS,1.0μg) was microinjected into bilateral RT. We found that sleep was decreased and wakefulness was increased compared with control; when L-Glu (0.2μg) was microinjected into DRN and methysergide (MS,1.0μg), a non-selective 5-HT antagonist, was microinjected into bilateral RT, We found that sleep was enhanced and wakefulness was reduced compared with L-Glu group.5.2 When p-chlorophenylalanine (PCPA, 10μg) was microinjected into DRN and NS (1.0μg) was microinjected into bilateral RT, We found that sleep was increased and wakefulness was decreased compared with control; microinjection of 5-hydroxytryptaphan (5-HTP, 1.0μg), which can convert to 5-HT by the enzyme tryptophane hydroxylase and enhance 5-HT into bilateral RT, could block the effect of microinjection of PCPA into DRN on sleep-wakefulness cycle.

本研究采用脑立体定位、核团插管、微量注射、多导睡眠描记等方法，研究丘脑网状核(nucleus reticularis thalami，RT)中γ-氨基丁酸(gamma-amino butyric acid ，GABA)能神经元、一氧化氮(nitrogen monoxidum，NO)能神经元、阿片肽类神经递质、环一磷酸腺苷(cyclic adenosine monophosphate，cAMP)及中缝背核(nucleus raphes dorsalis，DRN)至RT的5-羟色胺(5-hydroxytryptamine，5-HT)能神经纤维投射对大鼠睡眠-觉醒周期的影响及其作用机制。1 RT内GABA能神经元对大鼠睡眠-觉醒周期的影响1.1大鼠RT内微量注射GABA合成关键酶抑制剂3-巯基丙酸(3-MP，5μg)，注射当天睡眠时间略有减少，第二日睡眠时间显著减少，觉醒时间明显增多，第三、四日睡眠和觉醒时间逐渐恢复至正常。1.2大鼠RT内微量注射GABA受体激动剂GABA( 1.0μg)后，与生理盐水组比较，睡眠时间增加，觉醒时间减少；而RT内微量注射L-谷氨酸(glutamic acid， L-Glu， 0.2μg)后，睡眠时间减少，觉醒时间增加；RT内微量注射GABAA受体阻断剂荷包牡丹碱(bicuculline，BIC，1.0μg)后，睡眠时间减少，觉醒时间增加；RT内微量注射GABAB受体激动剂氯苯氨丁酸(baclofen，BAC，1.0μg)后，产生了与GABA相似的促睡眠效果。2 RT内NO能神经元对大鼠睡眠-觉醒周期的影响2.1大鼠RT内微量注射NO的前体L-精氨酸(L-Arg，0.5μg)后，与生理盐水组对比，睡眠时间略有减少，但无显著性意义；而RT内微量注射NO的供体硝普钠(Sodium Nitroprusside，SNP，0.2μg)后可明显增加觉醒时间，缩短睡眠时间；微量注射一氧化氮合酶抑制剂L-硝基精氨酸(L-arginine，L-NNA，2.0μg)后，引起睡眠时间增多，觉醒时间减少。2.2大鼠RT内同时微量注射L-NNA(2.0μg)和SNP(0.2μg)后与L-NNA组比较发现SNP逆转了L-NNA的促睡眠作用；RT内同时微量注射L-NNA(2.0μg)和L-Arg(0.5μg)后，与L-NNA(2.0μg)组比较发现L-Arg可以增加觉醒而缩短睡眠，其促觉醒作用未能被NOS的抑制剂L-NNA所逆转。3 RT内阿片肽对大鼠睡眠-觉醒周期的影响3.1大鼠RT内微量注射硫酸吗啡(morphine sulfate，MOR，1.0μg)后与生理盐水组对比，睡眠时间减少而觉醒时间增加； RT内微量注射阿片肽受体拮抗剂盐酸纳洛酮(naloxone hydrochloride，NAL，1.0μg)后与生理盐水组比较，睡眠时间增加而觉醒时间减少。3.2大鼠RT内同时微量注射MOR(1.0μg)和NAL(1.0μg)后，与生理盐水组对比，原有的MOR促觉醒效果和NAL的促睡眠效果都没有表现。4 RT内环一磷酸腺苷信使对大鼠睡眠-觉醒周期的影响大鼠RT内微量注射cAMP(1.0μg)后与NS(1.0μg)组比较，睡眠时间增多而觉醒时间减少；RT内微量注射亚甲蓝(methylene blue，MB，1.0μg)后，与NS组比较，睡眠时间增多而觉醒时间减少。5中缝背核投射到丘脑网状核的5-羟色胺能神经纤维对大鼠睡眠-觉醒周期的影响5.1大鼠DRN内微量注射L-Glu(0.2μg)，同时在双侧RT内微量注射NS (1.0μg)后，与对照组(DRN和双侧RT注射NS， 0.2μg)比较，睡眠时间减少，觉醒时间增多；大鼠DRN内微量注射L-Glu(0.2μg)，同时在双侧RT内微量注射二甲基麦角新碱(methysergide， MS， 1.0μg )后，与对照组(DRN注射L-Glu 0.2μg，双侧RT注射NS 1.0μg)比较，睡眠时间增多，觉醒时间减少。5.2大鼠DRN内微量注射对氯苯丙氨酸(p-chlorophenylalanine，PCPA，10μg)，同时在双侧RT内微量注射NS (1.0μg)后，与对照组(DRN和双侧RT注射NS， 1.0μg)比较，睡眠时间增多，觉醒时间减少；大鼠DRN内微量注射PCPA(10μg)，产生睡眠增多效应后，在双侧RT内微量注射5-羟色胺酸(5-hydroxytryptaphan ， 5-HTP， 1.0μg )后，与对照组(DRN注射PCPA 10μg，双侧RT注射NS 1.0μg)比较，睡眠时间减少，觉醒时间增多。

Thedisadvantages of the dragline are that must dig below itself, it connot dig with force into compacted terial, it cannot dig on steep slopes, and its dumping and digging are not accurate.

拉铲挖土机的缺点是只能挖比它本身低的土，不能施加压力挖入压实的土壤内，不能再陡坡上挖土，而且挖卸都不准确。

CHARGES. At this time DIG charges no brokerage fees or commissions. DIG may charge for incidental banking-related fees such as wire charges for deposits/withdrawals and returned check fees. DIG reserves the right to change its fee structure at any time without notice. Fees do not currency but may in the

27.2 客户同意由于客户未能完全和实时履行客户协议规定义务或因客户提供的信息不真实、不准确导致DIG受到的损失，造成的支出包括律师费用等向DIG做出赔偿。

I am dug ah dig, was to give many water first, I took water come out, continue to dig, next gouge a skeleton, I threw the death's head, continue to dig, discovered a copper mine again next, I throw taphole shovel, convert electric drill, continue to be gotten downward.

我挖呀挖，先是出了不少水，我把水抽了出来，继续挖，然后又挖出了一个骷髅，我把骷髅扔了上来，继续挖，然后又发现了一个铜矿，我扔出铁锨，改用电钻，继续往下钻。

Haemocyte membrane of Fenneropenaeus chinensis was purified and WSSV was labeled with DIG, the DIG-WSSV was incubated with MAb and the mixture was added to ELISA plate coated with membrane haemocyte, then the binding was detected with anti-DIG antibody.

提取中国对虾血细胞膜并对WSSV进行地高辛标记，将病毒和各单抗混合作用后吸附对虾血细胞膜，使用抗地高辛抗体检测结合到血细胞膜上的病毒。

John Sweeney of the 10th Armored Division tried to dig in,"but the ground was made up of heavy wet clay and our entrenching shovel couldn't dig into it."

第10装甲师约翰·斯文尼中士试图挖掘散兵坑，&地层是又湿又重的黏土，我们的军用铁锹根本就挖不下去。&

It is in order to dig golden bay mine exemple, analysed the difficulty that anile mine exists and contradiction, put forward to must be relied on deepen mechanism of reform, changeover, science and technology to dig go, synergism of billabong of aggrandizement interior management, executive come off sentry duty, depletion of numbers, development diversified economy and tertiary industy will solve these difficulty and problem, put forward to make the business changes the line of production smoothly the countermeasure of development.

以挖金湾矿为例，分析了衰老矿井存在的困难和矛盾，提出了必须依靠深化改革、转换机制、科技挖潜、强化内部治理、实施下岗分流、减员增效，发展多种经营和第三产业来解决这些困难和问题，并提出了使企业顺利转产发展的对策。

Dig and dig, some people are digging the oyster, some people are catching the fiddler crab, but I?

挖~挖挖，有人在挖蛤蛎，有人在抓招潮蟹，而我呢？

Included on the full-console versions are Pac-Man, Ms. Pac-Man, Galaga, Galaxian, Dig Dug, Pole Position, Pole Position II, Rolling Thunder, Rally X, Bosconian, Dragon Spirit, Sky Kid, Xevious, and Mappy, while the GBA version has a slightly more limited selection, but will include the big guns like Pac-Man and Dig Dug.

包括对充分控制台版本的PAC -男子，女士的PAC -男子，" Galaga "， galaxian ，挖挖，一极的地位，极立场，二，滚动打雷，凝聚X的bosconian ，龙的精神，天空的孩子， xevious ， mappy ，而的GBA版本有一个稍微比较有限的选择，但将包括大炮一样的PAC -男子及挖挖。

Although these two mountains are very high,they can't increase high again , dig to order would reduce a litter, why we dig the gravamen?

这两座山虽然很高，却不会再增高了，挖点就会少一点，为什么挖不平呢？

That's what makes this mod unique.

这也正是这个MOD独特。

I now feel furious and want to do something.

现在我觉得非常气愤，想做点什么。