이 연구는 중국 국립 자연 과학 재단(81973569, 82274207 및 82104533), 청두 중의학 대학의 Xinglin Scholar Research Promotion Project(XKTD2022013) 및 Ningxia의 주요 연구 개발 프로그램(2023BEG02012)의 지원을 받았습니다.

홍경 천은 중국 쓰촨성 간쯔 티베트족 자치현 간쯔현 주오다 설산(N 31.44570°, E 99.96086°, 4892m)에서 채취됩니다. 이 식물은 청두 전통 중국 의학 대학 민족 의학 학교의 Yi Zhang 교수에 의해 진품으로 인증되었습니다.
1. 홍경천 채취
<ol><li>Rhodiola crenulata의 서식지 지도를 촬영하십시오.</li>
	<li>Rhodiola crenulata의 전체 식물, 잎, 꽃받침 및 뿌리 줄기를 촬영하십시오.</li>
	<li>삽을 사용하여 Rhodiola crenulata 에서 1m 이내의 잡초와 부러진 돌을 제거하여 후속 원활한 채굴을 보장합니다.</li>
	<li>뿌리권 전체가 보일 때까지 괭이로 흙을 파내고 꼭지 뿌리를 모으십시오. 참고 : 약재에 사용되는 Rhodiola crenulata 의 뿌리와 뿌리 줄기는 꽃 줄기가 시들어가는 가을에 수집해야합니다.</li>
</ol>2. 특성 식별
<ol><li>육안으로 홍경 천의 외관 특성을 관찰하십시오 : 원통형의 짧은 꼭지 뿌리와 뿌리 줄기, 갈색 표면, 분홍색 패턴이있는 막색 표피, 주황색-빨간색 또는 부르고뉴 조각.</li>
	<li>냄새로 식별: 코 근처에 있을 때 향긋한 냄새를 맡습니다.</li>
	<li>맛으로 식별: 작은 뿌리 조각을 입에 넣고 먼저 한 모금 씹은 다음 약간 쓴 맛이 나고 그 다음에는 단맛이 납니다.</li>
</ol>3. 약용 분말에서 전분 과립의 현미경 식별
<ol><li>Rhodiola crenulata 표면의 흙을 브러시로 제거하고 40 °C의 오븐에 넣고 24 시간마다 허브를 뒤집습니다. 알림: 의약 재료의 깨지기 쉬운 것은 수분 건조의 표준으로 간주됩니다.</li>
	<li>분말 기계를 사용하여 건조된 약재를 분말화하고 약용 3호 체를 사용하여 약용 분말을 여과합니다( 재료 표 참조).</li>
	<li>깨끗한 슬라이드( 재료 표 참조)를 가져와서 해부 바늘로 분말을 파고( 재료 표 참조) 2mm 이내의 슬라이드의 1/3에 고르게 놓습니다.</li>
	<li>유리 스포이드( 재료 표 참조)를 사용하여 분말에 탈이온수 한 방울을 추가합니다. 핀셋( 재료 표 참조)을 사용하여 덮개 유리의 한쪽 끝( 재료 표 참조)을 잡고 액체 레벨을 빠르게 만지고 분말을 덮습니다. 알림: 해부학 바늘을 사용하여 물과 약용 분말을 부드럽게 혼합하여 균일한 샘플 혼합을 보장합니다. 슬라이드, 파우더 및 커버 유리 사이에 기포가 없어야 합니다.</li>
	<li>현미경을 열고( 재료 표 참조) 3.4단계의 슬라이드를 플랫폼에 놓아 고정합니다. 광원과 거친 초점 나선을 조정하여 분말을 확인합니다. 조직이 선명하게 보일 때까지 미세한 파라포컬 나선형을 조정합니다. 40x 대물렌즈로 전환하고 전분 과립을 관찰합니다. 알림: 단일 또는 다중 곡물로 나타나는 곡물을 녹말로 처리하면 탯줄이 헤링본 또는 균열 모양으로 나타납니다.</li>
</ol>4. 약용 분말의 카테터, 코르크 세포, 섬유, 목재 실질 세포 및 색소 덩어리의 현미경 식별
<ol><li>깨끗한 슬라이드( 재료 표 참조)를 가져와서 해부 바늘로 분말을 파고( 재료 표 참조) 슬라이드의 1/3에 놓습니다.</li>
	<li>유리 스포이드( 재료 표 참조)를 사용하여 분말에 클로랄 수화물 한 방울( 재료 표 참조)을 추가합니다. 핀셋으로 슬라이드를 가져 와서 ( 재료 표 참조) 알코올 램프에서 가열하십시오.amp 매번 1초 동안 세 번. 알림: 가열하는 동안 기포를 피해야 합니다. 액체는 흐르지 않는 상태로 유지되어 침투가 완료되었음을 나타냅니다.</li>
	<li>유리 스포이드를 사용하여 글리세린 한 방울을 추가합니다( 재료 표 참조). 핀셋을 사용하여 커버 유리의 한쪽 끝을 잡고 액체 레벨을 빠르게 만집니다.</li>
	<li>현미경을 열고 슬라이드를 플랫폼에 올려 고정합니다. 광원과 거친 초점 나선을 조정하여 분말을 확인합니다. 조직이 선명하게 보일 때까지 미세한 파라포컬 나선형을 조정합니다. 40x 대물 렌즈로 전환하여 카테터, 코르크 세포, 섬유, 목재 실질 세포 및 색소 블록을 관찰합니다. 참고: 코르크 세포의 다각형 또는 긴 다각형, 명백한 나선형 구조를 가진 나선형 용기, 옥살산 칼슘 모래 결정을 포함하는 물관부 실질, 적색 또는 갈색을 띤 적색 안료 블록.</li>
</ol>5. Rhodiola crenulata의 박층 크로마토그래피(TLC) 샘플 및 그 참조 준비
<ol><li>칭량 용지를 저울에 놓고( 재료 표 참조) Rhodiola crenulata 분말 3g의 무게를 잰다.</li>
	<li>분말을 100mL 원뿔형 병에 넣고 ( 재료 표 참조) 큰 배 피펫으로 25mL의 메탄올을 추가합니다 ( 재료 표 참조). 원뿔형 병을 초음파 기기에 넣습니다. 전력을 250W로, 주파수를 40kHz로, 시간을 30분으로 설정하고( 자료표 참조) 기기를 켭니다. 알림: 초음파 기기의 목적은 Rhodiola crenulata 의 분말이 후속 박막 크로마토그래피 실험 결과에 영향을 주지 않고 완전히 용해되도록 하는 것입니다.</li>
	<li>원뿔형 병을 제거하고 외부 병을 흐르는 물로 실온(RT)으로 헹굽니다.</li>
	<li>5.3단계에서 준비한 액체 800μL를 1mL 주사기로 흡입합니다. 0.22μm 미세 다공성 필터 멤브레인( 재료 표 참조)으로 필터링하고 크로마토그래피 샘플 병에 Rhodiola crenulata 의 미드스트림 샘플 용액 400μL를 수집합니다.</li>
	<li>2mg의 살리드로사이드, 티로졸 및 갈산( 재료 표 참조)을 각각 3개의 개별 100mL 원뿔형 병에 넣습니다. 큰 배 피펫이 있는 메탄올 25mL를 모든 원뿔형 병에 넣습니다.</li>
	<li>원뿔형 병을 초음파 기기에 넣고 전력을 250W로, 주파수를 40kHz로, 시간을 30분으로 설정하고 5.3단계를 반복합니다( 재료 표 참조).</li>
	<li>1mL 주사기로 5.6단계에서 준비한 액체 800μL를 흡입합니다. 0.22μm 미세 다공성 필터 멤브레인( 재료 표 참조)으로 필터링하고 해당 크로마토그래피 샘플 병에 400μL의 미드스트림 살리드로사이드, 티로졸 및 갈산 표준 용액을 수집합니다.</li>
</ol>6. TLC 식별
<ol><li>피펫 트리클로로메탄(5mL), 에틸 아세테이트(4mL), 메탄올(2mL) 및 포름산(0.5mL)( 재료 표 참조). 이중 탱크 크로마토그래피 실린더의 한쪽 면에 추가하고( 재료 표 참조) 흔든 후 골고루 섞습니다. 상부 실린더 헤드를 덮습니다.</li>
	<li>갈산, 살리드로사이드, 티로솔 표준 용액 및 홍경천 크레눌라타 용액을 A1-A4 위치의 샘플 랙에 놓습니다.</li>
	<li>5cm x 10cm 실리콘 시트( 재료 표 참조)를 샘플링 테이블에 놓습니다. 자동 s를 시작amp기계( 재료 표 참조)를 클릭하고 공기 제어 밸브를 엽니다.</li>
	<li>visionCATS 소프트웨어를 엽니다( 자료표 참조). 새로 만들기(New) > 새 폴더 ( Rhodiola crenulata sample test)를 클릭한 > 확인(OK)을 클릭합니다. New Method(이름 : Rhodiola crenulata sample test)를 클릭하고 OK > ATS 4> 클릭합니다.</li>
	<li>단계 정의 완료(Finish Step Definition)를 클릭합니다. Track Assignment(할당 추적)를 클릭하여 설명(gallic acid, salidroside, tyrosol 및 Rhodiola crenulata 샘플)을 편집합니다.</li>
	<li>HPTLC 단계를 클릭합니다. 얇은 레이어 매개 변수 (5cm x 10cm)를 설정합니다. 응용 프로그램 유형(밴드)을 선택하고 매개변수(표 1)를 설정한 다음 확인 버튼을 클릭합니다.</li>
	<li>충전/헹굼 품질을 엽니다. Fill only Programmed Volume(프로그래밍된 볼륨만 채우기)을 선택합니다. 바이알 바닥 레벨(mm)을 0.5로 설정하고 OK 버튼을 클릭합니다.</li>
	<li>Execute Method(메서드 실행)를 클릭합니다.</li>
	<li>Track Assignment 버튼을 클릭하고 Center를 선택한 다음 매개변수를 설정합니다(표 2).</li>
	<li>자동 샘플링을 위해 계속 버튼을 클릭합니다.</li>
	<li>자동 샘플링 기계와 공기 밸브를 끕니다. 자동 샘플링 기계에서 실리콘 시트를 제거합니다.</li>
	<li>실리콘 시트를 6.1단계에서 이중 탱크 크로마토그래피 실린더의 반대쪽에 넣고 상부 실린더 헤드를 덮고 20분 동안 사전 포화시킵니다.</li>
	<li>얇은 층판의 상단을 핀셋으로 부드럽게 고정하고 실리콘 시트를 현상제에 빠르게 넣고 상부 실린더 헤드를 덮습니다. 알림: 펼쳐지는 전면 가장자리가 얇은 층판의 상단에서 0.5-1.0cm 떨어져 있을 때 실리콘 시트를 꺼냅니다.</li>
	<li>유기 용매가 증발한 후 실온에서 실리콘 시트 표면에 발색 용액을 분사하여 발색 결과를 얻습니다. 참고 : 발색 용액은 2 % FeCl3 및 1 % K3 [Fe (CN) 6]를 포함하는 수용액입니다.</li>
</ol>

Rhodiola crenulate의 현미경 특성 분석 및 TLC 기반 식별

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K., G&#243;mez-Serranillos, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=DNA-based+authentication+and+metabolomics+analysis+of+medicinal+plants+samples+by+DNA+barcoding+and+ultra-high-performance+liquid+chromatography/triple+quadrupole+mass+spectrometry+(UHPLC-MS).">DNA-based authentication and metabolomics analysis of medicinal plants samples by DNA barcoding and ultra-high-performance liquid chromatography/triple quadrupole mass spectrometry (UHPLC-MS).</a> Plants (Basel, Switzerland). 9 (11), 1601 (2020).</li><li>. Rhodiola renulata (Hook.fil &amp; Thomson) H.Ohba Available from: <a target="_blank" href="https://www.gbif.org/species/4200529">https://www.gbif.org/species/4200529</a> (2023)</li><li>. Plant + data center Available from: <a target="_blank" href="https://www.iplant.cn/">https://www.iplant.cn/</a> (2023)</li><li>. Chinese virtual herbarium Available from: <a target="_blank" href="https://www.cvh.ac.cn/">https://www.cvh.ac.cn/</a> (2023)</li><li>Cunningham, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=There+&amp;#34;ain't+no+mountain+high+enough&amp;#34;?:+The+drivers,+diversity+and+sustainability+of+China's+Rhodiola+trade.">There &amp;#34;ain't no mountain high enough&amp;#34;?: The drivers, diversity and sustainability of China's Rhodiola trade.</a> Journal of Ethnopharmacology. 252, 112379 (2020).</li><li>Wang, Q., Ruan, X., Jin, Z. H., Yan, Q. C., Tu, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+Rhodiola+species+by+using+RP-HPLC.">Identification of Rhodiola species by using RP-HPLC.</a> Journal of Zhejiang University. Science. B. 6 (6), 477-482 (2005).</li><li>Dong, X., Guo, Y., Xiong, C., Sun, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+two+major+Rhodiola+species+and+the+systemic+changing+characteristics+of+metabolites+of+Rhodiola+crenulata+in+different+altitudes+by+chemical+methods+combined+with+UPLC-QqQ-MS-based+metabolomics.">Evaluation of two major Rhodiola species and the systemic changing characteristics of metabolites of Rhodiola crenulata in different altitudes by chemical methods combined with UPLC-QqQ-MS-based metabolomics.</a> Molecules. 25 (18), 4062 (2020).</li><li>Zhao, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HPLC+fingerprint+differentiation+between+natural+and+ex+situ+populations+of+Rhodiola+sachalinensis+from+Changbai+Mountain,+China.">HPLC fingerprint differentiation between natural and ex situ populations of Rhodiola sachalinensis from Changbai Mountain, China.</a> PloS One. 9 (11), 112869 (2014).</li><li>Yu, Y. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quality+evaluation+of+Tibetan+medicine+Rhodiolae+crenulatae+radix+et+rhizome+based+on+sensory+recognition+and+near+infrared+spectrum+analysis.">Quality evaluation of Tibetan medicine Rhodiolae crenulatae radix et rhizome based on sensory recognition and near infrared spectrum analysis.</a> Chengdu University of Traditional Chinese Medicine. , (2020).</li><li>Lv, X. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simultaneous+determination+of+5+chemical+components+in+3+kinds+of+Rhodiola+rosea+by+HPLC.">Simultaneous determination of 5 chemical components in 3 kinds of Rhodiola rosea by HPLC.</a> China Pharmacy. 29 (18), 2515-2519 (2018).</li><li>Dong, T., Sha, Y., Liu, H., Sun, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Altitudinal+variation+of+metabolites,+mineral+elements+and+antioxidant+activities+of+Rhodiola+crenulata+(Hook.f.+&amp;+Thomson)+H.Ohba.">Altitudinal variation of metabolites, mineral elements and antioxidant activities of Rhodiola crenulata (Hook.f. &amp; Thomson) H.Ohba.</a> Molecules. 26 (23), 7383 (2021).</li><li>Ma, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+UHPLC+fingerprints+combined+with+chemical+pattern+recognition+analysis+in+the+differentiation+of+six+Rhodiola+species.">Application of UHPLC fingerprints combined with chemical pattern recognition analysis in the differentiation of six Rhodiola species.</a> Molecules. 26 (22), 6855 (2021).</li><li>Li, X. 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저자는 공개할 것이 없습니다.

세계에는 90 종 이상의 홍경천 식물이 있으며 모든 종의 60 % 이상이 중국에서 발견되며 홍경천, 홍경천, 홍경천 achalinensis 및 홍경천 17을 포함한 일반적인 식물이 있습니다. 중국 약전(2020)의 첫 번째 부분에 기록된 홍경천은 높은 고도에서 재배되는 티베트 전통 약재입니다. 홍경천에 대한 시장 수요는 매년 증가하고 있으므로 공급원의 올바른 ?...

약초는 중국에서 오랜 역사와 풍부한 응용 경험을 가지고 있으며 Shennong의 약초 고전1에서 최초의 체계적인 기록이었습니다. 말라리아에 적용된 아르테미시닌의 발견은 약초의 새로운 단계1로의 발전을 촉진했습니다. 현대 과학 기술을 사용하여 약초의 정확한 메커니즘을 밝혀냄으로써 약초의 이용률과 수요가 증가하여 약초에 대한 새로운 국제 시장이 열렸습니다 2,3,4. 그러나 이것은 일련의 부정적인 영향으로 이어졌습니다. 비전문가는 한약의 특성에 대해 막연하게 이해하고 있어 한약의 사용이 큰 안전 위험에 직면하게 됩니다5.
홍경천의 식물 중 하나인 홍경천은 주로 티베트, 윈난 북서부, 중국 쓰촨성 서부에 분포한다(그림 1)6,7. 홍경천은 살리드로사이드, 티로졸, 갈산 및 기타 화합물로 구성되어 있으며, "기를 활성화하고 혈액 순환을 촉진하며 맥박을 맑게 하고 천식을 진정시키는 기능"8,9,10,11 기능을 통해 저산소증 관련 질병을 치료합니다. 현장 조사에 따르면 홍경천은 고도 4,000-5,600m의 고산 거골 지대, 협곡 경사면 및 암석 틈새에서 발견 될 수 있습니다. 성장 환경은 춥고 햇빛과 강렬한 방사선으로 가득 차 있으며 고산 초원 생태계에 속합니다. Rhodiola crenulata는 성장 지형에 따라 라멜라 및 점형 개체군에 분포할 수 있으며, 유전자 흐름은 교차 수분을 통해 수행될 수 있습니다.
홍경천 속의 꽃가루 낙태, 불법 발굴 및 퇴화 된 생태 환경은 홍경천 crenulata를 멸종 위기에 처한 종으로 만듭니다 6,12. Rhodiola crenulata의 높은 약용 가치를 고려할 때 위조 제품이 시장에 유입될 것으로 예상됩니다. 이 기사에서는 Rhodiola crenulata의 서식지와 몇 가지 편리한 실험실 식별 방법을 제시합니다. 첫째, 홍경천의 생육 환경과 약효를 관찰하였다. 둘째, 약용 분말의 미세 구조를 현미경으로 관찰하였다. 마지막 단계가 핵심입니다. Rhodiola crenulata의 대표 성분은 특정 물질에서 이들 성분의 상이한 흡착 또는 용해 특성에 따라 분리되고 확인되었습니다. 약용 식물의 DNA 기반 인증 또는 대사체학 분석 방법은 복잡하고 비용이 많이 듭니다13. 이러한 기본적이고 편리하며 경제적인 방법으로 홍경천을 신속하게 식별할 수 있습니다.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.22 &#956;m millipore filter</td>
			<td>Millipore</td>
			<td>SLGP033RB</td>
			<td></td>
		</tr>
		<tr>
			<td>Automatic sampling machine</td>
			<td>CAMAG</td>
			<td>ATS 4</td>
			<td></td>
		</tr>
		<tr>
			<td>Chloral hydrate</td>
			<td>Fuzhou Brunei Technology Co., Ltd</td>
			<td>ST1002</td>
			<td></td>
		</tr>
		<tr>
			<td>Chromatographic&#160;sample&#160;bottles</td>
			<td>Zhejiang ALWSCI Technology Co., Ltd</td>
			<td>C0000008</td>
			<td></td>
		</tr>
		<tr>
			<td>Conical flask</td>
			<td>Sichuan Shubo Co., Ltd</td>
			<td>1121</td>
			<td></td>
		</tr>
		<tr>
			<td>Cover glass</td>
			<td>Citotest Labware Manufacturing Co., Ltd</td>
			<td>10211818c</td>
			<td></td>
		</tr>
		<tr>
			<td>Dissecting needle</td>
			<td>Shanghai Bingyu Fluid Technology Co., Ltd</td>
			<td>BY-5026</td>
			<td></td>
		</tr>
		<tr>
			<td>Electronic balance</td>
			<td>SHIMADZU</td>
			<td>ATX124</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethyl acetate</td>
			<td>Chengdu Kelong Chemical Co., Ltd</td>
			<td>2022120901</td>
			<td></td>
		</tr>
		<tr>
			<td>Formic acid</td>
			<td>Chengdu Kelong Chemical Co., Ltd</td>
			<td>2021110801</td>
			<td></td>
		</tr>
		<tr>
			<td>Gallic acid</td>
			<td>Chengdu Herbpurify Co., Ltd</td>
			<td>M-017</td>
			<td></td>
		</tr>
		<tr>
			<td>Glycerol</td>
			<td>Sinopharm Chemical Reagent Co., Ltd</td>
			<td>10010618</td>
			<td></td>
		</tr>
		<tr>
			<td>High speed&#160; crusher</td>
			<td>Beijing Zhongxingweiye Instrument Co., Ltd</td>
			<td>FW-100</td>
			<td></td>
		</tr>
		<tr>
			<td>Methanol</td>
			<td>Chengdu Kelong Chemical Co., Ltd</td>
			<td>20230108</td>
			<td></td>
		</tr>
		<tr>
			<td>Microscope</td>
			<td>Chongqing Oprec Nistrument Co.,&#160; Ltd</td>
			<td>B203</td>
			<td></td>
		</tr>
		<tr>
			<td>Microscope slide</td>
			<td>Citotest Labware Manufacturing Co., Ltd</td>
			<td>7105P-G</td>
			<td></td>
		</tr>
		<tr>
			<td>Oven</td>
			<td>Shanghai Yuejin Medical Equipment Co., Ltd</td>
			<td>DHG-8145</td>
			<td></td>
		</tr>
		<tr>
			<td>Pharmacopoeia sieve</td>
			<td>Hangzhou Xingrun sieve factory</td>
			<td>572423281330</td>
			<td></td>
		</tr>
		<tr>
			<td>Pipette</td>
			<td>Changde BKMAM Biotechnology Co., Ltd</td>
			<td>120302008</td>
			<td></td>
		</tr>
		<tr>
			<td>Salidroside</td>
			<td>Chengdu Herbpurify Co., Ltd</td>
			<td>H-040</td>
			<td></td>
		</tr>
		<tr>
			<td>Saturate tank&#160;</td>
			<td>Yancheng Liegu Technology Co., Ltd</td>
			<td>10*20 P-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Silica gel plate</td>
			<td>Yantai Jiangyou Silica Gel Development&#160; Co., Ltd</td>
			<td>HSG20211227</td>
			<td></td>
		</tr>
		<tr>
			<td>Trichloromethane</td>
			<td>Chengdu Kelong Chemical Co., Ltd</td>
			<td>20221013-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Tweezer&#160;</td>
			<td>Changde BKMAM Biotechnology Co., Ltd</td>
			<td>130302027</td>
			<td></td>
		</tr>
		<tr>
			<td>Tyrosol</td>
			<td>Chengdu Herbpurify Co., Ltd</td>
			<td>L-042</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrasound equipment</td>
			<td>Ningbo Xinyi Ultrasonic Equipment Co., Ltd</td>
			<td>SB-8200DTS</td>
			<td></td>
		</tr>
		<tr>
			<td>Volumetric pipet</td>
			<td>Changde BKMAM Biotechnology Co., Ltd</td>
			<td>120301006</td>
			<td></td>
		</tr>
	</tbody>
</table>

field collection and laboratory routine identification of rhodiola crenulata

의약품 성분의 식별은 의약품 안전성의 전제이자 보증입니다. 과학 연구자들의 대다수는 약초의 간단하고, 빠르고, 효과적이고, 저렴한 식별 과정을 선호할 수밖에 없습니다. 홍경천은 높은 고도에서 자라는 티베트 전통 약재로 주로 중국의 티베트, 윈난, 쓰촨 지역에 분포합니다. 홍경천은 항염증, 항저산소증, 항산화 등 여러 생체 활성을 가지고 있으며 발전 가능성이 큽니다. 시장 수요가 증가하고 자원 함량이 급격히 감소함에 따라 홍경천 crenulata 의 많은 혼란스러운 제품이 사람들을 괴롭히고 있습니다. 따라서 이 프로토콜은 일상적인 실험실 테스트와 결합된 현장에서 Rhodiola crenulata 를 식별하기 위한 표준 프로세스를 도입합니다. 서식지, 미세한 특징 및 박층 크로마토그래피의 조합은 의심할 여지 없이 홍경 천을 빠르고 효율적이며 경제적으로 식별하여 티베트 의학의 지속적인 발전과 의약 재료의 품질 관리에 기여할 것입니다.

이 실험 프로토콜은 현장에서 Rhodiola crenulate 를 식별하고 수집하는 방법을 설명합니다. Rhodiola crenulate 는 고산 거골 지역, 협곡 경사면 및 높은 고도의 암석 틈새에 서식하는 경향이 있습니다. Rhodiola crenulate 의 서식지, 전체 식물, 꽃 및 잎은 그림 2에 나와 있습니다. Rhodiola crenulate 는 적갈색의 뿌리줄기를 가지고 있습니다(그림 3A</str...

Watch this Scientific Journal Video about Field Collection and Laboratory Routine Identification of Rhodiola crenulata at JoVE.com

저자 스포트라이트: Rhodiola crenulata 에 대한 식별 방법 개발 및 자원 분포 및 약리학적 효과 조사

여기에서는 서식지, 식물 형태, 의약 특성, 미세한 특징 및 박층 크로마토그래피에서 Rhodiola Crenulata 의 식별에 대해 설명합니다.

Rhodiola crenulata의 현장 수집 및 실험실 일상적인 식별

The identification of medicinal materials is the premise and guarantee of drug safety. The majority of scientific researchers are bound to favor the ...

본 연구의 범위는 민족 의학에 대한 자원 조사를 수행하는 데 중점을 두고 있습니다. 특히, 본 연구는 홍경천의 식별을 위한 빠르고 편리한 방법을 개발하는 것을 목표로 한다. 우리가 제공하는 프로토콜은 다양한 약초 재료를 식별하기 위한 비용 효율적이고 간단하며 신속한 방법을 제시합니다.우리 실험실은 두 가지 주요 영역, 즉 홍경천과 관련된 연구의 분포 및 이 종의 약리학적 영향을 조사하는 데 향후 연구 노력을 집중할 것입니다.

field-collection-laboratory-routine-identification-rhodiola

Research

JoVE Journal

Medicine

Field Collection and Laboratory Routine Identification of Rhodiola crenulata

521 조회수.  Chengdu University of Traditional Chinese Medicine. 본 연구의 범위는 민족 의학에 대한 자원 조사를 수행하는 데 중점을 두고 있습니다. 특히, 본 연구는 홍경천의 식별을 위한 빠르고 편리한 방법을 개발하는 것을 목표로 한다. 우리가 제공하는 프로토콜은 다양한 약초 재료를 식별하기 위한 비용 효율적이고 간단하며 신속한 방법을 제시합니다.우리 실험실은 두 가지 주요 영역, 즉 홍경천과 관련된 연구의 분포 및 이 ?...

비디오: 저자 스포트라이트: Rhodiola crenulata 에 대한 식별 방법 개발 및 자원 분포 및 약리학적 효과 조사

Collection Protocol for Human Pancreas

This dissection and sampling procedure was developed for the Network for Pancreatic Organ Donors with Diabetes (nPOD) program to standardize preparation of pancreas recovered from cadaveric organ donors. The pancreas is divided into 3 main regions (head, body, tail) followed by serial transverse sections throughout the medial to lateral axis. Alternating sections are used for fixed paraffin and fresh frozen blocks and remnant samples are minced for snap frozen sample preparations, either with or without RNAse inhibitors, for DNA, RNA, or protein isolation. The overall goal of the pancreas dissection procedure is to sample the entire pancreas while maintaining anatomical orientation.
Endocrine cell heterogeneity in terms of islet composition, size, and numbers is reported for human islets compared to rodent islets 1. The majority of human islets from the pancreas head, body and tail regions are composed of insulin-containing &beta;-cells followed by lower proportions of glucagon-containing &alpha;-cells and somatostatin-containing &delta;-cells. Pancreatic polypeptide-containing PP cells and ghrelin-containing epsilon cells are also present but in small numbers. In contrast, the uncinate region contains islets that are primarily composed of pancreatic polypeptide-containing PP cells 2. These regional islet variations arise from developmental differences. The pancreas develops from the ventral and dorsal pancreatic buds in the foregut and after rotation of the stomach and duodenum, the ventral lobe moves and fuses with the dorsal 3. The ventral lobe forms the posterior portion of the head including the uncinate process while the dorsal lobe gives rise to the rest of the organ. Regional pancreatic variation is also reported with the tail region having higher islet density compared to other regions and the dorsal lobe-derived components undergoing selective atrophy in type 1 diabetes4,5.
Additional organs and tissues are often recovered from the organ donors and include pancreatic lymph nodes, spleen and non-pancreatic lymph nodes. These samples are recovered with similar formats as for the pancreas with the addition of isolation of cryopreserved cells. When the proximal duodenum is included with the pancreas, duodenal mucosa may be collected for paraffin and frozen blocks and minced snap frozen preparations.

This video demonstrates a dissection procedure for processing human pancreas into multiple storage formats. Anatomical orientation is maintained throughout the pancreatic regions to allow definition of regional islet composition and density.

인간의 췌장을위한 컬렉션 프로토콜

This dissection and sampling procedure was developed for the Network for Pancreatic Organ Donors with Diabetes (nPOD) program to standardize preparation ...

연구

의학

The Use of Primary Human Fibroblasts for Monitoring Mitochondrial Phenotypes in the Field of Parkinson's Disease

Parkinson's disease (PD) is the second most common movement disorder and affects 1% of people over the age of 60 1. Because ageing is the most important risk factor, cases of PD will increase during the next decades 2. Next to pathological protein folding and impaired protein degradation pathways, alterations of mitochondrial function and morphology were pointed out as further hallmark of neurodegeneration in PD 3-11.
After years of research in murine and human cancer cells as in vitro models to dissect molecular pathways of Parkinsonism, the use of human fibroblasts from patients and appropriate controls as ex vivo models has become a valuable research tool, if potential caveats are considered. Other than immortalized, rather artificial cell models, primary fibroblasts from patients carrying disease-associated mutations apparently reflect important pathological features of the human disease.
Here we delineate the procedure of taking skin biopsies, culturing human fibroblasts and using detailed protocols for essential microscopic techniques to define mitochondrial phenotypes. These were used to investigate different features associated with PD that are relevant to mitochondrial function and dynamics. Ex vivo, mitochondria can be analyzed in terms of their function, morphology, colocalization with lysosomes (the organelles degrading dysfunctional mitochondria) and degradation via the lysosomal pathway. These phenotypes are highly relevant for the identification of early signs of PD and may precede clinical motor symptoms in human disease-gene carriers. Hence, the assays presented here can be utilized as valuable tools to identify pathological features of neurodegeneration and help to define new therapeutic strategies in PD.

The Use of Primary Human Fibroblasts for Monitoring Mitochondrial Phenotypes in the Field of Parkinson&#039;s Disease

Fibroblasts from patients carrying mutations in Parkinson's disease-causing genes represent an easily accessible ex vivo model to study disease-associated phenotypes. Live cell imaging gives the opportunity to study morphological and functional parameters in living cells. Here we describe the preparation of human fibroblasts and subsequent monitoring of mitochondrial phenotypes.

파킨슨 병 분야에서 Mitochondrial Phenotypes을 모니터링하기위한 기본 인간 섬유 아세포의 사용

Parkinson's disease (PD) is the second most  common movement disorder and affects 1% of people over the age of 60 1. Because ageing is  the most important ...

Localization, Identification, and Excision of Murine Adipose Depots

Obesity has increased dramatically in the last few decades and affects over one third of the adult US population. The economic effect of obesity in 2005 reached a staggering sum of $190.2 billion in direct medical costs alone. Obesity is a major risk factor for a wide host of diseases. Historically, little was known regarding adipose and its major and essential functions in the body. Brown and white adipose are the two main types of adipose but current literature has identified a new type of fat called brite or beige adipose. Research has shown that adipose depots have specific metabolic profiles and certain depots allow for a propensity for obesity and other related disorders. The goal of this protocol is to provide researchers the capacity to identify and excise adipose depots that will allow for the analysis of different factorial effects on adipose; as well as the beneficial or detrimental role adipose plays in disease and overall health. Isolation and excision of adipose depots allows investigators to look at gross morphological changes as well as histological changes. The adipose isolated can also be used for molecular studies to evaluate transcriptional and translational change or for in vitro experimentation to discover targets of interest and mechanisms of action. This technique is superior to other published techniques due to the design allowing for isolation of multiple depots with simplicity and minimal contamination.

Due to the drastic and negative connection between obesity and other comorbidities, research on the role adipose plays in disease and overall health is warranted. We present a protocol for the isolation and excision of adipose depots allowing for the study of adipose using in situ and in vitro methods.

현지화, 식별 및 절제 쥣과 지방이 종점의

Obesity has increased dramatically in the last few decades and affects over one third of the adult US population. The economic effect of obesity in 2005 ...

Methodology for Sputum Induction and Laboratory Processing

The technique of sputum induction and processing is a recognized non-invasive method allowing the collection and analysis of cells from the airways, which is interesting in various respiratory diseases like asthma, chronic obstructive pulmonary disease (COPD), chronic cough, or idiopathic pulmonary fibrosis. This technique is well tolerated, safe and non-invasive, but is currently limited to research services and specialized centers in clinical practice because it is technically demanding, time-consuming, and requires trained staff. The success rate of sputum induction and analysis is about 80%.
Here, we describe the induction and laboratory processing of sputum samples. Sputum is induced by inhalation of hypertonic or isotonic saline with salbutamol. For the processing, we use the whole sputum technique. Dithiothreitol (DTT) is used to allow mucolysis of sputum samples. The primary aim of sputum processing is to obtain a differential cell count to study the cell types present in the airway lumen. Additional analyses may also be performed on sputum supernatant and sputum cells, which may allow further investigation into inflammatory processes and immune mechanisms. Examples include studying mediators in sputum supernatant and performing a large spectrum of analysis on sputum cells such as flow cytometry, genomics, or proteomics.
Finally, representative results of sputum analysis in healthy controls, asthmatics, and COPD patients are presented.

Here we describe the technique of sputum induction. This protocol also explains the sputum processing to perform a differential cell count and to collect sputum supernatant and cells for further analysis.

선 유도 및 실험실 방법론

The technique of sputum induction and processing is a recognized non-invasive method allowing the collection and analysis of cells from the airways, which ...

Routine Screening Method for Microparticles in Platelet Transfusions

Platelet inventory management based on screening microparticle content in platelet concentrates is a new quality improvement initiative for hospital blood banks. Cells fragment off microparticles (MP) when they are stressed. Blood and blood components may contain cellular fragments from a variety of cells, most notably from activated platelets. When performing their roles as innate immune cells and major players in coagulation and hemostasis, platelets change shape and generate microparticles. With dynamic light scattering (DLS)-based microparticle detection, it is possible to differentiate activated (high microparticle) from non-activated (low microparticle) platelets in transfusions, and optimize the use of this scarce blood product. Previous research suggests that providing non-activated platelets for prophylactic use in hematology-oncology patients could reduce their risk of becoming refractory and improve patient care. The goal of this screening method is to routinely differentiate activated from non-activated platelets. The method described here outlines the steps to be performed for routine platelet inventory management in a hospital blood bank: obtaining a sample from a platelet transfusion, loading the sample into the capillary for DLS measurement, performing the DLS test to identify microparticles, and using the reported microparticle content to identify activated platelets.

Platelet inventory management based on screening microparticle content in platelet concentrates is a new quality improvement initiative in hospital blood banks. The goal is to differentiate activated from non-activated platelets to optimize the use of platelets. Providing non-activated platelets to hematology-oncology patients might reduce their high risk to become refractory.

혈소판 수혈에 미에 대 한 메서드를 검사 하는 루틴

Platelet inventory management based on screening microparticle content in platelet concentrates is a new quality improvement initiative for hospital blood ...

Remote Laboratory Management: Respiratory Virus Diagnostics

An uptick in recent pandemics (Ebola, Zika, MERS, influenza, etc.) underlines the need for a more 'nimble,' coordinated response that addresses a multitude of issues ranging from transportation, access, facilities, equipment, and communication to provider training. To address this need, we have developed an innovative, scalable, logistics-enhanced, mobile, laboratory facility for emergencies and epidemics in resource-constrained global settings. Utilizing a background in clinical operations as an academic medical center, we designed a rapidly-deployable, modular BSL-2 and BSL-3 facility with user-friendly software for tracking and management of drugs and supplies in remote regions during epidemics and outbreaks. Here, we present our intermodal, mobile, expandable shipping-container laboratory units. The design of the laboratory facilitates off-grid usage by minimizing power consumption and allowing alternate water sources. The unit's information communication technology (ICT) platform provides (i) user-friendly tablet-based documentation, (ii) enhanced tracking of patients and supplies, and (iii) integrated communication onsite with built-in telehealth capabilities. To ensure quality in remote environments, we have developed a checklist for a basic laboratory workflow and a protocol for respiratory viral diagnosis using reverse-transcription polymerase chain reaction (RT-PCR). As described, this innovative and comprehensive approach allows for the provision of laboratory capability in resource-limited global environments.

A rapidly-deployable, off-grid laboratory has been designed and built for remote, resource-constrained global settings. The features and critical aspects of the logistically-enhanced, expandable, multifunctional laboratory modules are explored. A checklist for a basic laboratory workflow and a protocol for a respiratory viral diagnostic test are developed and presented.

원격 실험실 관리: 호흡기 바이러스 진단

An uptick in recent pandemics (Ebola, Zika, MERS, influenza, etc.) underlines the need for a more 'nimble,' coordinated response that addresses a ...

Learning Modern Laryngeal Surgery in a Dissection Laboratory

Surgery for laryngeal malignancies requires millimetric accuracy from the different endoscopic and open techniques available. Practice of this surgery is almost completely reserved to a few referral centers that deal with a large proportion of this pathology. Practice on human specimens is not always possible for ethical, economic, or availability reasons. The aim of this study is to provide a reproducible method for the organization of a laryngeal laboratory on ex vivo animal models where it is possible to approach, learn, and refine laryngeal techniques. Porcine and ovine larynges are ideal, affordable, models to simulate laryngeal surgery given their similarity to the human larynx in their anatomical layout and tissue composition. Herein, the surgical steps of transoral laser surgery, open partial horizontal laryngectomy, and total laryngectomy are reported. The merging of endoscopic and exoscopic views guarantees an inside-out perspective, which is vital for the comprehension of the complex laryngeal anatomy. The method was successfully adopted during three sessions of a dissection course &#34;Lary-Gym&#34;. Further perspectives on robotic surgical training are described.

The purpose of this paper is to illustrate how to organize a reproducible laboratory for laryngeal surgery on affordable and closely similar animal laryngeal models in order to improve anatomical and surgical knowledge and skills.

해부 실험실에서 현대 후두 수술 배우기

Surgery for laryngeal malignancies requires millimetric accuracy from the different endoscopic and open techniques available. Practice of this surgery is ...

An In Vitro Dissolution Determination of Multi-Index Components in Tibetan Medicine Rhodiola Granules

The composition of the Tibetan medicine Rhodiola granules (RG) is complex, and the overall quality of RG is difficult to determine. Therefore, establishing a method to determine the multi-component in vitro dissolution of RG is of great significance for quality control. This study uses the second paddle method of the fourth general rule 0931 from the Chinese Pharmacopoeia (2020 edition), compliant with apparatus 2 of the United States Pharmacopeia (USP). The dissolution apparatus was set to a rotation speed of 100 rpm with ultrapure water as the dissolution medium. A sample volume of 1 mL was collected at each timepoint. Furthermore, the cumulative dissolution of gallic acid, salidroside, and ethyl gallic acid in RG at different time points was determined by high-performance liquid chromatography (HPLC). Finally, the dissolution curves were drawn, and the curves were fitted to the GompertzMod, the Gompertz, the Logistic, and the Weibull equations. The results showed that the cumulative dissolution of gallic acid in RG was over 80% at 1 min, the cumulative dissolution of salidroside and ethyl gallic acid was over 65% at 5 min, and the cumulative dissolution of each index component decreased after 30 min. The curve fitting demonstrated that the GompertzMod equation was the best-fitting model for each index component of RG. In conclusion, the dissolution test method described in this protocol is simple, accurate, and reliable. It can characterize the dissolution behavior of the index components in RG in vitro, which provides a methodological reference for quality control of RG and quality evaluation of other ethnic compounds.

An In Vitro Dissolution Determination of Multi-Index Components in Tibetan Medicine Rhodiola Granules

Here, we test the dissolution of Rhodiola granules (RG) in vitro, draw dissolution curves of salidroside, gallic acid, and ethyl gallate in ultrapure water, and fit the curves to different mathematical models. This protocol provides information and guidance for in vivo bioequivalence and in vivo-in vitro correlation studies of RG.

The composition of the Tibetan medicine Rhodiola granules (RG) is complex, and the overall quality of RG is difficult to determine. Therefore, ...

Ferric Chloride-Induced Arterial Thrombosis and Sample Collection for 3D Electron Microscopy Analysis

Cardiovascular diseases are a leading cause of mortality and morbidity worldwide. Aberrant thrombosis is a common feature of systemic conditions like diabetes and obesity, and chronic inflammatory diseases like atherosclerosis, cancer, and autoimmune diseases. Upon vascular injury, usually the coagulation system, platelets, and endothelium act in an orchestrated manner to prevent bleeding by forming a clot at the site of the injury. Abnormalities in this process lead to either excessive bleeding or uncontrolled thrombosis/insufficient antithrombotic activity, which translates into vessel occlusion and its sequelae. The FeCl3-induced carotid injury model is a valuable tool in probing how thrombosis initiates and progresses in vivo. This model involves endothelial damage/denudation and subsequent clot formation at the injured site. It provides a highly sensitive, quantitative assay to monitor vascular damage and clot formation in response to different degrees of vascular damage. Once optimized, this standard technique can be used to study the molecular mechanisms underlying thrombosis, as well as the ultrastructural changes in platelets in a growing thrombus. This assay is also useful to study the efficacy of antithrombotic and antiplatelet agents. This article explains how to initiate and monitor FeCl3-induced arterial thrombosis and how to collect samples for analysis by electron microscopy.

The present protocol describes how to use a FeCl3-mediated injury to induce arterial thrombosis, and how to collect and prepare arterial injury samples at various stages of thrombosis for electron microscopy analysis.

염화제이철 유발 동맥 혈전증 및 3D 전자 현미경 분석을 위한 시료 수집

Cardiovascular diseases are a leading cause of mortality and morbidity worldwide. Aberrant thrombosis is a common feature of systemic conditions like ...

인간 limbal niche 세포를 분리, 배양 및 식별하기 위한 프로토콜

Isolation and Identification of Limbal Niche Cells

Here we report a standard procedure for the isolation and identification of limbal niche cells (LNCs). Limbus tissue obtained from an eye bank was used for LNCs isolation. The tissue was divided into 12 pieces under aseptic conditions and digested for 18 h at 37 &#176;C in the cell culture incubator using collagenase A to obtain cell clusters with LNCs and limbal&#160;epithelial progenitor cells. The cell clusters were further digested for 15 min at 37 &#176;C using 0.25% trypsin-EDTA to obtain single cells and then cultured in modified embryonic stem cell medium (MESCM) on a plastic surface coated with 5% Matrigel. Cells were passaged upon 70% confluence, and LNCs were identified using immunofluorescence, real-time quantitative PCR (qPCR), and flow cytometry. Primary LNCs were isolated and passaged more than 12 times. The proliferation activity of LNCs from P4 to P6 was the highest. LNCs expressed higher stem cell markers than BMMSCs (SCF, Nestin, Rex1, SSEA4, CD73, CD90, MSX1, P75NTR, and PDGFR&#946;). Furthermore, results showed that P4 LNCs uniformly expressed VIM, CD90, CD105, and PDGFR&#946;, but not Pan-CK, which could be used as a marker for the identification of LNCs. Flow cytometric analysis showed that approximately 95%, 97%, 92%, and 11% of LNCs expressed CD73, CD90, CD105, and SCF respectively, while they were&#160;68%, 99%, 20%, and 3% in&#160;BMMSCs. The standard process for LNC&#160;isolation and identification could provide&#160;a reliable laboratory basis for the widespread use of LNCs.

저자 스포트라이트: 광범위한 LNC 연구를 지원하기 위해 LNC(Limbal Niche Cell) 분리 및 특성 분석 표준화 

Here, we present a protocol to isolate and identify the human limbal niche cells.

Limbal Niche 세포의 분리 및 식별

Here we report a standard procedure for the isolation and identification of limbal niche cells (LNCs). Limbus tissue obtained from an eye bank was used ...