Methods A nested RT-PCR was used for detecting one fragment of SARS-CoV RNA in oropharyngeal swabs from 3 SARS probable patients ,4 SARS suspect patients and other 27 patients with fever in Hangzhou, and the nested RT-PCR product from one SARS probable patient was sequenced.Meanwhile in these 3 SARS probable patients, other three RT-PCR methods, including a hemi -nested RT-PCR targeted for another fragment of SARS-CoV RNA, a real-time RT -PCR and a modified nested real-time RT-PCR, were employed to detect SARS-Co V RNA.Results Two positives were found in the 3 SARS pro bable patients, and none positive in 4 SARS suspect patients and other 27 patien ts with fever, using the nested RT-PCR.

方法對2003年杭州市3例嚴重急性呼吸綜合征臨床確診患者人人爽人人干人人操人人、4例疑似和27名醫(yī)學觀察者的咽拭子標本用巢式實時RT-PCR檢測SARS-CoV核酸欧美日韩视频在线，對陽性擴增產(chǎn)物進行核酸序列測定1024手机看片日韩人妻，同時對3例患者的標本應用半巢式RT-PCR檢測另一位點SARS-CoV核酸片段，并應用常規(guī)實時RT-PCR技術(shù)及改良實時RT-PCR技術(shù)進行檢測岛国性爱视频在线。

There were only 5 cases negative for SYT-SSX genomic by seminested PCR, which were positive for SYT-SSX mRNA by RT-PCR. Three cases of synovial sarcoma negative for SYT-SSX mRNA were also negative for SYT-SSX genomic DNA. One case of synovial sarcoma was positive for SYT-SSX1 genomic DNA by seminested PCR, which can not be distinguished for SYT-SSX fusion type by RT-PCR.

只有5例SYT-SSX mRNA呈陽性而半巢式PCR SYT-SSX DNA檢測呈陰性。3例RT-PCR檢測SYT-SSX mRNA呈陰性者半巢式PCR亦為陰性結(jié)果欧美孕交丰满熟妇XXXX性。1例RT-PCR方法無法確定融合基因類型的病例SYT-SSX DNA檢測為SYT-SSX1型日本免费的黄色网站。

Temporal and spatial expression pattern of gibel carp pou2 during embryogenesis was investigated by RT-PCR and whole-mount in situ hybridization. RT-PCR result indicates that there is maternal transcript of gibel carp pou2 in early embryos and the zygotic pou2 transcript is abundantly expressed from blastula stage. Large amount of pou2 transcript is detected at both 50% and 90% epiboly stages. At 100% epiboly stage, however, the pou2 transcript reduces rapidly and disappears at the later embryogenesis. Whole-mount in situ hybridization shows that the maternal transcript ubiquitously exists in all blastomere cells.

我們用RT-PCR和整體原位雜交的方法研究了銀鯽pou2基因在胚胎發(fā)育過程中的時空表達圖式RT-PCR結(jié)果顯示，銀鯽pou2基因有母源轉(zhuǎn)錄本男人天堂网手机在线视频，其合子基因在高囊胚期強烈表達，在50%下包期和90%下包期也有高量的轉(zhuǎn)錄本黄色夜里自己看的视频，但在100%下包期表達量急劇降低淫荡乱女，至體節(jié)期時已經(jīng)完全檢測不到其轉(zhuǎn)錄本胚胎整體原位雜交結(jié)果顯示其母源轉(zhuǎn)錄本在所有的胚盤細胞中。

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金德管業(yè)集團引進德國尤尼克塑料設(shè)備制造公司鋁塑復合管生產(chǎn)線亚洲最大黄色视频高清免费在线观看、德國克勞斯·馬菲公司PVC-U給排水管生產(chǎn)線黄色片一道本、德國巴頓菲爾德公司全塑管生產(chǎn)線及香港震雄注塑機，采用高科技手段生產(chǎn)"金德"牌 PAP鋁塑復合管欧美精品videosseⅹ少妇、PPR鋁塑管思思热在线观看视频网站、PE-X交聯(lián)聚乙烯管、PE-RT耐高溫聚乙烯管无码黄色毛片、PPR全塑管亚洲日韩欧美在线视频播放、PVC-U新型復合排水管、PVC-U給水管久久人人操人人爽、電纜護套管等和與之配套的銅管件欧美性爱电影免费看、PPR管件、PP-RT管件一级绝黄A片AAA、 PVC管件及相應的安裝工具等免费99精品久久七七桃花。

Maffei Corporation PVC-U drainage pipes to the production line, the German company Barton Rumsfeld plastic production line and the Hong Kong Chen Hsong injection molding machine and other advanced production equipment, use of hi-tech means of production "Jinde" brand aluminum PAP composite pipe, PPR aluminum-plastic tube, PE-X cross-linked polyethylene pipe, PE-RT polyethylene pipe temperature, PPR plastic pipes, PE gas pipe, PE water pipe, PE mining pipe, HDPE large caliber double-wall corrugated pipe, HDPE Large Diameter Winding Pipe, PVC-U new composite pipes, PVC-U to the water pipes, cable sheathing, etc., and be compatible with the brass pieces, PPR pipe fittings, PE-RT pipe, PE water supply pipe pieces, PE gas pipe pieces, PE mining pipe pieces, PVC pipe fittings and the corresponding installation tools.

瑪菲公司PVC-U給排水管生產(chǎn)線、德國巴頓菲爾德公司全塑生產(chǎn)線及香港震雄注塑機等先進生產(chǎn)設(shè)備啪啪免费特级黄片，采用高科技手段生產(chǎn)"金德"牌PAP鋁塑復合管熟女啪啪视频，PPR鋁塑管、PE-X交聯(lián)聚乙烯管你懂网站在线观看、PE—RT耐高溫聚乙烯管精美黄色网站、PPR全塑管、PE燃氣管能观看的三级国产tv视频网址、PE給水管免费色色网页、PE礦用管、HDPE大口徑雙壁波紋管直播欧美牲爱、HDPE大口徑纏繞管97免费在线视频观看、PVC—U新型復合排水管、PVC—U給水管白白在线播放免费视频、電纜護套管等和與之配套的銅管件在线不卡高清无码、PPR管件黄色免站视频、PE—RT管件、PE給水管管件欧美成人情欲小说视频在线、PE燃氣管管件欧美又粗又长啪啪动态、PE礦用管管件、PVC管件及相應的安裝工具等国产一级黄片乱伦。

Methods The MAL gene expression in 45 cancer patients was detected with RT-PCR and semi-quantitative RT-PCR technique.

采用RT-PCR和半定量RT-PCR方法檢測45例食管癌組織和癌旁食管黏膜組織中MAL基因的表達黄色A片wwwcom。

RESULT: 1.Ouabain act on lens sodium-pump,under the LM,lens anterior capsular membrane discontinuous,epithelium cells clustered,occluding zonule seperated,lens fiber layers fractured.Under the EM,cells totally hollowed,mitochondria swelling,myelin figure appeared.RT-PCR examine the expression condition of αsubunit of sodium pump on mRNA level,α1、α2 and α3-isoform are all decreased.2.Digoxin act on lens sodium-pump,under the LM,lens cell oedema,linkage distructed,extensive exfoliation.Under the EM,plasma appeared little half-transparant hollow region,mitochondria swelling and ridge disappeared. RT-PCR examine,α1深夜色网址大全、α2 and α3-isoform are all decreased.3.Amphotericin B act on lens sodium-pump,under the LM,lens epithelium cells linked tightly,arranged in-line,lens fiber layers arranged tightly and regularily.Under the EM,abbundant cellular organes,exuberant cells function indicated. RT-PCR examine the expression condition of αsubunit of sodium pump on mRNA level,α1 and α3-isoform are increased significantly,demonstrated isoform-specific action.4D-thyroxine act on lens sodium-pump,under the LM,lens plasmalemma integrated,cells arranged tightly and regularily.Under the EM,nucleus fission appeared,desmosome half-desmosome and tensile microfilaments linked the cells. RT-PCR examine,α2 and α3-isoform are increased, also demonstrated isoform-specific action.5.Vitamin E act on lens sodium-pump,under the LM,lens anterior capsular membrane continuous and smooth,epithelium cells tightly linked,lens fiber layers appearede hollow region occasionally.Under the EM,lateral membrane high density belt appeared,abundant nucleolus. RT-PCR examine,onlyα1-isoform are increased, demonstrated significantly isoform-specific action.6.DMSO act on lens sodium-pump,under the LM,lens anterior capsular membrane slightly thicker,cells linkage partly distructed.Under the EM,plasmalemma denaturation,mitochondria swelling.RT-PCR examine,α1日操夜操人人操、α2 and α3-isoform are all altered slightly and haven't significant meanning.

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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.

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Methods: Oocytes in the GV stage were separated from ovary by squeezing method. In mouse germinal vesicle GV stage, the expression of ATP8 gene in the mitochondria in the single oocyte was detected by RT-PCR, in which, cDNA was synthesized with two methods: one was the single GV-stage oocyte directly to be placed RT, the other was to perform RT after eliminating mtDNA and nucleus DNA with the EeoR Ⅰ enzyme and Dnase. And the product of RT-PCR was cloned and sequenced.

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