High-Throughput Screening to Identify Plant Derived Human LDH-A Inhibitors
Identifieur interne : 000742 ( Pmc/Curation ); précédent : 000741; suivant : 000743High-Throughput Screening to Identify Plant Derived Human LDH-A Inhibitors
Auteurs : S. Deiab [États-Unis] ; E. Mazzio [États-Unis] ; S. Messeha [États-Unis] ; N. Mack [États-Unis] ; K. F. A. Soliman [États-Unis]Source :
- European journal of medicinal plants [ 2231-0894 ] ; 2013.
Abstract
Lactate dehydrogenase (LDH)-A is highly expressed in diverse human malignant tumors, parallel to aggressive metastatic disease, resistance to radiation/chemotherapy and clinically poor outcome. Although this enzyme constitutes a plausible target in treatment of advanced cancer, there are few known LDH-A inhibitors.
In this work, we utilized a high-throughput enzyme micro-array format to screen and evaluate > 900 commonly used medicinal plant extracts (0.00001-.5
The protein sequence of purified enzyme was confirmed using 1D gel electrophoresis- MALDI
Of the natural extracts tested, the lowest IC50s [<0.001 mg/ml] were obtained by: Chinese Gallnut (
These findings reflect relative potency by rank of commonly used herbs and plants that contain human LDH-A inhibitory properties. Future research will be required to isolate chemical constituents within these plants responsible for LDH-A inhibition and investigate potential therapeutic application.
Url:
PubMed: 24478981
PubMed Central: 3903096
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">High-Throughput Screening to Identify Plant Derived Human LDH-A Inhibitors</title>
<author><name sortKey="Deiab, S" sort="Deiab, S" uniqKey="Deiab S" first="S." last="Deiab">S. Deiab</name>
<affiliation wicri:level="1"><nlm:aff id="A1">College of Pharmacy and Pharmaceutical Sciences, Florida A & M University, Tallahassee, Florida 32307, USA</nlm:aff>
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<wicri:regionArea>College of Pharmacy and Pharmaceutical Sciences, Florida A & M University, Tallahassee, Florida 32307</wicri:regionArea>
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<author><name sortKey="Mazzio, E" sort="Mazzio, E" uniqKey="Mazzio E" first="E." last="Mazzio">E. Mazzio</name>
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<wicri:regionArea>College of Pharmacy and Pharmaceutical Sciences, Florida A & M University, Tallahassee, Florida 32307</wicri:regionArea>
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<author><name sortKey="Messeha, S" sort="Messeha, S" uniqKey="Messeha S" first="S." last="Messeha">S. Messeha</name>
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<wicri:regionArea>College of Pharmacy and Pharmaceutical Sciences, Florida A & M University, Tallahassee, Florida 32307</wicri:regionArea>
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<author><name sortKey="Soliman, K F A" sort="Soliman, K F A" uniqKey="Soliman K" first="K. F. A." last="Soliman">K. F. A. Soliman</name>
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<wicri:regionArea>College of Pharmacy and Pharmaceutical Sciences, Florida A & M University, Tallahassee, Florida 32307</wicri:regionArea>
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<series><title level="j">European journal of medicinal plants</title>
<idno type="eISSN">2231-0894</idno>
<imprint><date when="2013">2013</date>
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<front><div type="abstract" xml:lang="en"><sec id="S1"><title>Aims</title>
<p id="P1">Lactate dehydrogenase (LDH)-A is highly expressed in diverse human malignant tumors, parallel to aggressive metastatic disease, resistance to radiation/chemotherapy and clinically poor outcome. Although this enzyme constitutes a plausible target in treatment of advanced cancer, there are few known LDH-A inhibitors.</p>
</sec>
<sec id="S2"><title>Study Design</title>
<p id="P2">In this work, we utilized a high-throughput enzyme micro-array format to screen and evaluate > 900 commonly used medicinal plant extracts (0.00001-.5 <italic>mg/ml</italic>
) for capacity to inhibit activity of recombinant full length human LDHA; EC .<italic>1.1.1.27</italic>
.</p>
</sec>
<sec id="S3"><title>Methodology</title>
<p id="P3">The protein sequence of purified enzyme was confirmed using 1D gel electrophoresis- MALDI<italic>-TOF-</italic>
MS/MS, enzyme activity was validated by oxidation of NADH (500μM) and kinetic inhibition established in the presence of a known inhibitor (Oxalic Acid).</p>
</sec>
<sec id="S4"><title>Results</title>
<p id="P4">Of the natural extracts tested, the lowest IC<sub>50</sub>
s [<0.001 mg/ml] were obtained by: Chinese Gallnut (<italic>Melaphis chinensis gallnut)</italic>
, Bladderwrack (<italic>Fucus vesiculosus)</italic>
, Kelp (<italic>Laminaria Japonica</italic>
) and Babul (<italic>Acacia Arabica)</italic>
. Forty-six additional herbs contained significant LDH-A inhibitory properties with IC<sub>50</sub>
s [<0.07 mg/ml], some of which have common names of Arjun, Pipsissewa, Cinnamon, Pink Rose Buds/Petals, Wintergreen, Cat’s Claw, Witch Hazel Root and Rhodiola Root.</p>
</sec>
<sec id="S5"><title>Conclusion</title>
<p id="P5">These findings reflect relative potency by rank of commonly used herbs and plants that contain human LDH-A inhibitory properties. Future research will be required to isolate chemical constituents within these plants responsible for LDH-A inhibition and investigate potential therapeutic application.</p>
</sec>
</div>
</front>
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</TEI>
<pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<pmc-dir>properties manuscript</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-journal-id">101583475</journal-id>
<journal-id journal-id-type="pubmed-jr-id">40208</journal-id>
<journal-id journal-id-type="nlm-ta">European J Med Plants</journal-id>
<journal-id journal-id-type="iso-abbrev">European J Med Plants</journal-id>
<journal-title-group><journal-title>European journal of medicinal plants</journal-title>
</journal-title-group>
<issn pub-type="epub">2231-0894</issn>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">24478981</article-id>
<article-id pub-id-type="pmc">3903096</article-id>
<article-id pub-id-type="manuscript">NIHMS543685</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Article</subject>
</subj-group>
</article-categories>
<title-group><article-title>High-Throughput Screening to Identify Plant Derived Human LDH-A Inhibitors</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Deiab</surname>
<given-names>S.</given-names>
</name>
<xref ref-type="aff" rid="A1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Mazzio</surname>
<given-names>E.</given-names>
</name>
<xref ref-type="aff" rid="A1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Messeha</surname>
<given-names>S.</given-names>
</name>
<xref ref-type="aff" rid="A1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Mack</surname>
<given-names>N.</given-names>
</name>
<xref ref-type="aff" rid="A1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Soliman</surname>
<given-names>K. F. A.</given-names>
</name>
<xref ref-type="aff" rid="A1">1</xref>
<xref rid="FN1" ref-type="author-notes">*</xref>
</contrib>
</contrib-group>
<aff id="A1"><label>1</label>
College of Pharmacy and Pharmaceutical Sciences, Florida A & M University, Tallahassee, Florida 32307, USA</aff>
<author-notes><corresp id="FN1"><label>*</label>
Corresponding author: <email>karam.soliman@famu.edu</email>
;</corresp>
</author-notes>
<pub-date pub-type="nihms-submitted"><day>6</day>
<month>1</month>
<year>2014</year>
</pub-date>
<pub-date pub-type="epub"><year>2013</year>
</pub-date>
<pub-date pub-type="pmc-release"><day>27</day>
<month>1</month>
<year>2014</year>
</pub-date>
<volume>3</volume>
<issue>4</issue>
<fpage>603</fpage>
<lpage>615</lpage>
<permissions><copyright-statement>© 2013 Deiab et al.;</copyright-statement>
<copyright-year>2013</copyright-year>
<license license-type="open-access" xlink:href="http://creativecommons.org/licenses/by/3.0"><license-p>This is an Open Access article distributed under the terms of the Creative Commons Attribution License (<ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/3.0">http://creativecommons.org/licenses/by/3.0</ext-link>
), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</license-p>
</license>
</permissions>
<abstract><sec id="S1"><title>Aims</title>
<p id="P1">Lactate dehydrogenase (LDH)-A is highly expressed in diverse human malignant tumors, parallel to aggressive metastatic disease, resistance to radiation/chemotherapy and clinically poor outcome. Although this enzyme constitutes a plausible target in treatment of advanced cancer, there are few known LDH-A inhibitors.</p>
</sec>
<sec id="S2"><title>Study Design</title>
<p id="P2">In this work, we utilized a high-throughput enzyme micro-array format to screen and evaluate > 900 commonly used medicinal plant extracts (0.00001-.5 <italic>mg/ml</italic>
) for capacity to inhibit activity of recombinant full length human LDHA; EC .<italic>1.1.1.27</italic>
.</p>
</sec>
<sec id="S3"><title>Methodology</title>
<p id="P3">The protein sequence of purified enzyme was confirmed using 1D gel electrophoresis- MALDI<italic>-TOF-</italic>
MS/MS, enzyme activity was validated by oxidation of NADH (500μM) and kinetic inhibition established in the presence of a known inhibitor (Oxalic Acid).</p>
</sec>
<sec id="S4"><title>Results</title>
<p id="P4">Of the natural extracts tested, the lowest IC<sub>50</sub>
s [<0.001 mg/ml] were obtained by: Chinese Gallnut (<italic>Melaphis chinensis gallnut)</italic>
, Bladderwrack (<italic>Fucus vesiculosus)</italic>
, Kelp (<italic>Laminaria Japonica</italic>
) and Babul (<italic>Acacia Arabica)</italic>
. Forty-six additional herbs contained significant LDH-A inhibitory properties with IC<sub>50</sub>
s [<0.07 mg/ml], some of which have common names of Arjun, Pipsissewa, Cinnamon, Pink Rose Buds/Petals, Wintergreen, Cat’s Claw, Witch Hazel Root and Rhodiola Root.</p>
</sec>
<sec id="S5"><title>Conclusion</title>
<p id="P5">These findings reflect relative potency by rank of commonly used herbs and plants that contain human LDH-A inhibitory properties. Future research will be required to isolate chemical constituents within these plants responsible for LDH-A inhibition and investigate potential therapeutic application.</p>
</sec>
</abstract>
<kwd-group><kwd>Lactic acid</kwd>
<kwd>Warburg</kwd>
<kwd>LDH-A</kwd>
<kwd>inhibitor</kwd>
<kwd>herbs</kwd>
<kwd>medicinal plants</kwd>
</kwd-group>
</article-meta>
</front>
<floats-group><fig id="F1" orientation="portrait" position="float"><label>Fig. 1</label>
<caption><p>Fig. 1a. Human LDH-A Activity-time and enzyme concentration dependent NADH oxidation in the presence of 3mM pyruvate. The data represent μM NADH reduced from 0–75 minutes (incubation at RT) and are presented as the Mean ± S.E.M, n=4. Significance of difference for product formation between Time <sub>0</sub>
vs Time <sub>15 – 75 minutes</sub>
were determined using a two-way ANOVA.* p< 0.05</p>
<p>Fig. 1b. <italic>Human</italic>
LDH Activity at .02 Units/ml with time dependent NADH oxidation in the presence of 3mM pyurvate. The data represent μM NADH and are presented as the Mean ± S.E.M, n=4. Significance of difference for product formation between Time <sub>0</sub>
vs Time <sub>75 minutes</sub>
was determined using a one-way ANOVA followed by a Tukey post hoc test. * p<0 .05</p>
<p>Fig. 1c. Human LDH Activity – Inhibitor Control. The data represent % Enzyme Activity @ 75 Minutes and are presented as the Mean ± S.E.M, n=4. Significance of difference for enzyme activity between the control and oxalic acid (0.9–11.3 mM) was determined using a one-way ANOVA followed by a Tukey post hoc test. * p<0 .05</p>
</caption>
<graphic xlink:href="nihms543685f1a"></graphic>
<graphic xlink:href="nihms543685f1b"></graphic>
</fig>
<fig id="F2" orientation="portrait" position="float"><label>Fig. 2</label>
<caption><p>Mascot results for protein identification by peptide mass fingerprinting of Human LDH-A tryptic digest analyzed by MALDI-TOF/TOF-MS</p>
</caption>
<graphic xlink:href="nihms543685f2"></graphic>
</fig>
<fig id="F3" orientation="portrait" position="float"><label>Fig. 3</label>
<caption><p>A high-throughput enzyme experimental micro-array design. 905 extracts were evaluated for capacity to inhibit <italic>Human</italic>
LDH-A. A first tier screening was conducted at a final working concentration of 0.5 mg/ml for each herbal extract. Enzyme activity was continuously monitored over a 75 min period. Extracts demonstrating an IC<sub>50</sub>
<0.5 mg/ml (red dotted line -----) were screened through subsequent tier evaluations</p>
</caption>
<graphic xlink:href="nihms543685f3"></graphic>
</fig>
<fig id="F4" orientation="portrait" position="float"><label>Fig. 4</label>
<caption><p>Most Potent Herbal Extract Inhibitors of <italic>Human</italic>
LDH-A activity. The data represent LDH-A activity as % control in the presence or absence of extracts and are presented as Mean ± S.E.M., n=4. IC<sub>50</sub>
concentrations were established from a sigmoidal fit dose-response equation and significance of difference between the controls vs. treatment was determined using a one-way ANOVA followed by a Tukey post hoc test. * p<0.05</p>
</caption>
<graphic xlink:href="nihms543685f4"></graphic>
</fig>
<table-wrap id="T1" position="float" orientation="landscape"><label>Table 1</label>
<caption><p>Natural source aqueous extracts with human LDH-A inhibitory potency by rank</p>
</caption>
<table frame="hsides" rules="groups"><thead><tr><th valign="bottom" align="right" rowspan="1" colspan="1">Rank</th>
<th valign="bottom" align="center" rowspan="1" colspan="1">ID No.</th>
<th valign="bottom" align="left" rowspan="1" colspan="1">Common ± Chinese Name</th>
<th valign="bottom" align="left" rowspan="1" colspan="1"><italic>Scientific Name</italic>
</th>
<th valign="bottom" align="center" rowspan="1" colspan="1">IC 50 mg/ml</th>
<th valign="bottom" align="center" rowspan="1" colspan="1">IC 50 μg/ml</th>
</tr>
</thead>
<tbody><tr><td align="right" valign="top" rowspan="1" colspan="1">1</td>
<td align="center" valign="top" rowspan="1" colspan="1">M22</td>
<td align="left" valign="top" rowspan="1" colspan="1">Wu Bei Zi</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Melaphis chinensis gallnut</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.00009</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.09</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">4</td>
<td align="center" valign="top" rowspan="1" colspan="1">B53</td>
<td align="left" valign="top" rowspan="1" colspan="1">Babul Chall Bark</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Acacia arabica</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.00075</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.75</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">2</td>
<td align="center" valign="top" rowspan="1" colspan="1">F17</td>
<td align="left" valign="top" rowspan="1" colspan="1">Bladderwrack</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Fucus vesiculosus</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.00100</td>
<td align="center" valign="top" rowspan="1" colspan="1">1.00</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">3</td>
<td align="center" valign="top" rowspan="1" colspan="1">K06</td>
<td align="left" valign="top" rowspan="1" colspan="1">Kelp Powder</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Laminaria Japonica</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.00134</td>
<td align="center" valign="top" rowspan="1" colspan="1">1.34</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">5</td>
<td align="center" valign="top" rowspan="1" colspan="1">B13</td>
<td align="left" valign="top" rowspan="1" colspan="1">Bayberry Root bark</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Morella cerifera</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.00158</td>
<td align="center" valign="top" rowspan="1" colspan="1">1.58</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">6</td>
<td align="center" valign="top" rowspan="1" colspan="1">C8</td>
<td align="left" valign="top" rowspan="1" colspan="1">CraneSbill Root</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Geranium maculatum</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.00173</td>
<td align="center" valign="top" rowspan="1" colspan="1">1.73</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">7</td>
<td align="center" valign="top" rowspan="1" colspan="1">A78</td>
<td align="left" valign="top" rowspan="1" colspan="1">Arjun</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Terminalia arjuna</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.00183</td>
<td align="center" valign="top" rowspan="1" colspan="1">1.83</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">8</td>
<td align="center" valign="top" rowspan="1" colspan="1">P54</td>
<td align="left" valign="top" rowspan="1" colspan="1">Ye Jiao Ten</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Polygonum multiflorum vine</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.00288</td>
<td align="center" valign="top" rowspan="1" colspan="1">2.88</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">9</td>
<td align="center" valign="top" rowspan="1" colspan="1">P56</td>
<td align="left" valign="top" rowspan="1" colspan="1">Mu Dan Pi</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Paeonia suffructicose root - bark</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.00298</td>
<td align="center" valign="top" rowspan="1" colspan="1">2.98</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">10</td>
<td align="center" valign="top" rowspan="1" colspan="1">W2</td>
<td align="left" valign="top" rowspan="1" colspan="1">Witch Hazel Root</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Hamamelis virginiana</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.00321</td>
<td align="center" valign="top" rowspan="1" colspan="1">3.21</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">11</td>
<td align="center" valign="top" rowspan="1" colspan="1">P82</td>
<td align="left" valign="top" rowspan="1" colspan="1">Pipsissewa</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Chimaphila umbellata</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.00410</td>
<td align="center" valign="top" rowspan="1" colspan="1">4.10</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">12</td>
<td align="center" valign="top" rowspan="1" colspan="1">C82</td>
<td align="left" valign="top" rowspan="1" colspan="1">Cinnamon powder</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Cinnamon powder</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.00434</td>
<td align="center" valign="top" rowspan="1" colspan="1">4.34</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">13</td>
<td align="center" valign="top" rowspan="1" colspan="1">R19</td>
<td align="left" valign="top" rowspan="1" colspan="1">Rose Buds and Petals Pink</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Rosa Rugosa Flower</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.00478</td>
<td align="center" valign="top" rowspan="1" colspan="1">4.78</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">14</td>
<td align="center" valign="top" rowspan="1" colspan="1">C10</td>
<td align="left" valign="top" rowspan="1" colspan="1">Cat Claw Bark</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Uncaria tomentosa</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.00483</td>
<td align="center" valign="top" rowspan="1" colspan="1">4.83</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">15</td>
<td align="center" valign="top" rowspan="1" colspan="1">D10</td>
<td align="left" valign="top" rowspan="1" colspan="1">Dryopteris Male Fern Rhizome</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Dryopteris crassirhizoma</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.00636</td>
<td align="center" valign="top" rowspan="1" colspan="1">6.36</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">16</td>
<td align="center" valign="top" rowspan="1" colspan="1">R25</td>
<td align="left" valign="top" rowspan="1" colspan="1">Rhodiola Root</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Rhodiola kirilowii</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.00809</td>
<td align="center" valign="top" rowspan="1" colspan="1">8.09</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">17</td>
<td align="center" valign="top" rowspan="1" colspan="1">T30</td>
<td align="left" valign="top" rowspan="1" colspan="1">Turkey Rhubarb</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Rheum palmatum</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.00958</td>
<td align="center" valign="top" rowspan="1" colspan="1">9.58</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">18</td>
<td align="center" valign="top" rowspan="1" colspan="1">G26</td>
<td align="left" valign="top" rowspan="1" colspan="1">Wintergreen</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Gaultheria procumbens</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.01086</td>
<td align="center" valign="top" rowspan="1" colspan="1">10.86</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">19</td>
<td align="center" valign="top" rowspan="1" colspan="1">XT76</td>
<td align="left" valign="top" rowspan="1" colspan="1">Longon Peel</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Dimocarpus longan</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.01195</td>
<td align="center" valign="top" rowspan="1" colspan="1">11.95</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">20</td>
<td align="center" valign="top" rowspan="1" colspan="1">H12</td>
<td align="left" valign="top" rowspan="1" colspan="1">Gloryvine Stem</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Sargentodoxa cuneata vine</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.01260</td>
<td align="center" valign="top" rowspan="1" colspan="1">12.60</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">21</td>
<td align="center" valign="top" rowspan="1" colspan="1">M6</td>
<td align="left" valign="top" rowspan="1" colspan="1">Meadowsweet Powder</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Filipendula ulmaria</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.01566</td>
<td align="center" valign="top" rowspan="1" colspan="1">15.66</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">22</td>
<td align="center" valign="top" rowspan="1" colspan="1">N2</td>
<td align="left" valign="top" rowspan="1" colspan="1">Neem Leaf</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Azadirachta indica</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.01836</td>
<td align="center" valign="top" rowspan="1" colspan="1">18.36</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">23</td>
<td align="center" valign="top" rowspan="1" colspan="1">T25</td>
<td align="left" valign="top" rowspan="1" colspan="1">Sang Ji Sheng</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Taxillus chinensis stem & leaf</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.02060</td>
<td align="center" valign="top" rowspan="1" colspan="1">20.60</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">24</td>
<td align="center" valign="top" rowspan="1" colspan="1">C32</td>
<td align="left" valign="top" rowspan="1" colspan="1">Cynomorium songaricum</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Cynomorium songaricum</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.02150</td>
<td align="center" valign="top" rowspan="1" colspan="1">21.50</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">25</td>
<td align="center" valign="top" rowspan="1" colspan="1">P55</td>
<td align="left" valign="top" rowspan="1" colspan="1">Chi Shao</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Paeonia Lactiflora</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.02283</td>
<td align="center" valign="top" rowspan="1" colspan="1">22.83</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">26</td>
<td align="center" valign="top" rowspan="1" colspan="1">P83</td>
<td align="left" valign="top" rowspan="1" colspan="1">Pygeum Bark</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Pygeum africanum</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.02623</td>
<td align="center" valign="top" rowspan="1" colspan="1">26.23</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">27</td>
<td align="center" valign="top" rowspan="1" colspan="1">P8</td>
<td align="left" valign="top" rowspan="1" colspan="1">Hu Zhang</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Polygonum cuspidatum rhizome</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.02842</td>
<td align="center" valign="top" rowspan="1" colspan="1">28.42</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">28</td>
<td align="center" valign="top" rowspan="1" colspan="1">XT 8</td>
<td align="left" valign="top" rowspan="1" colspan="1">Green Tea</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Camellia sinensis</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.02852</td>
<td align="center" valign="top" rowspan="1" colspan="1">28.52</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">29</td>
<td align="center" valign="top" rowspan="1" colspan="1">XT91</td>
<td align="left" valign="top" rowspan="1" colspan="1">Longon stem</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Dimocarpus longan</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.02973</td>
<td align="center" valign="top" rowspan="1" colspan="1">29.73</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">30</td>
<td align="center" valign="top" rowspan="1" colspan="1">A1</td>
<td align="left" valign="top" rowspan="1" colspan="1">Agrimony</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Agrimonia eupatoria</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.03072</td>
<td align="center" valign="top" rowspan="1" colspan="1">30.72</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">31</td>
<td align="center" valign="top" rowspan="1" colspan="1">L46</td>
<td align="left" valign="top" rowspan="1" colspan="1">Linden Leaf</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Tilia europaea</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.03152</td>
<td align="center" valign="top" rowspan="1" colspan="1">31.52</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">32</td>
<td align="center" valign="top" rowspan="1" colspan="1">A63</td>
<td align="left" valign="top" rowspan="1" colspan="1">Sha Ren Guang</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Amomum villosum fruit Shelled</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.04040</td>
<td align="center" valign="top" rowspan="1" colspan="1">40.40</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">33</td>
<td align="center" valign="top" rowspan="1" colspan="1">W16</td>
<td align="left" valign="top" rowspan="1" colspan="1">White Willow Bark</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Salix alba</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.04108</td>
<td align="center" valign="top" rowspan="1" colspan="1">41.08</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">34</td>
<td align="center" valign="top" rowspan="1" colspan="1">W15</td>
<td align="left" valign="top" rowspan="1" colspan="1">White Oak Bark</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Quercus alba</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.04154</td>
<td align="center" valign="top" rowspan="1" colspan="1">41.54</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">35</td>
<td align="center" valign="top" rowspan="1" colspan="1">G40</td>
<td align="left" valign="top" rowspan="1" colspan="1">Guarana Seed</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Paullinia cupana</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.04253</td>
<td align="center" valign="top" rowspan="1" colspan="1">42.53</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">36</td>
<td align="center" valign="top" rowspan="1" colspan="1">S11</td>
<td align="left" valign="top" rowspan="1" colspan="1">St Johns Wort</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>St Johns Wort</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.04408</td>
<td align="center" valign="top" rowspan="1" colspan="1">44.08</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">37</td>
<td align="center" valign="top" rowspan="1" colspan="1">L25</td>
<td align="left" valign="top" rowspan="1" colspan="1">Lychee Pit</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Litchi chinesis seed</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.04592</td>
<td align="center" valign="top" rowspan="1" colspan="1">45.92</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">38</td>
<td align="center" valign="top" rowspan="1" colspan="1">H4</td>
<td align="left" valign="top" rowspan="1" colspan="1">Hawthorne Leaf and Flower</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Crataegus laevigata</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.04790</td>
<td align="center" valign="top" rowspan="1" colspan="1">47.90</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">39</td>
<td align="center" valign="top" rowspan="1" colspan="1">P13</td>
<td align="left" valign="top" rowspan="1" colspan="1">Bian u</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Polygonum aviculare herb</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.05264</td>
<td align="center" valign="top" rowspan="1" colspan="1">52.64</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">40</td>
<td align="center" valign="top" rowspan="1" colspan="1">N1</td>
<td align="left" valign="top" rowspan="1" colspan="1">Nutmeg</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Myristica fragans</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.05811</td>
<td align="center" valign="top" rowspan="1" colspan="1">58.11</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">41</td>
<td align="center" valign="top" rowspan="1" colspan="1">S8</td>
<td align="left" valign="top" rowspan="1" colspan="1">Saw Palmetto Berry</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Serenoa repens</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.06608</td>
<td align="center" valign="top" rowspan="1" colspan="1">66.08</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">42</td>
<td align="center" valign="top" rowspan="1" colspan="1">M38</td>
<td align="left" valign="top" rowspan="1" colspan="1">Maiden Hair Fern</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Adiantum capillus</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.06847</td>
<td align="center" valign="top" rowspan="1" colspan="1">68.47</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">43</td>
<td align="center" valign="top" rowspan="1" colspan="1">T4</td>
<td align="left" valign="top" rowspan="1" colspan="1">Truja twigs, c/s</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Thuja occidentalis Cupressaceae</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.07676</td>
<td align="center" valign="top" rowspan="1" colspan="1">76.76</td>
</tr>
<tr><td align="right" valign="top" rowspan="1" colspan="1">44</td>
<td align="center" valign="top" rowspan="1" colspan="1">P29</td>
<td align="left" valign="top" rowspan="1" colspan="1">Wei Ling Cai</td>
<td align="left" valign="top" rowspan="1" colspan="1"><italic>Potentilla chinensis herb</italic>
</td>
<td align="center" valign="top" rowspan="1" colspan="1">0.07772</td>
<td align="center" valign="top" rowspan="1" colspan="1">77.72</td>
</tr>
</tbody>
</table>
</table-wrap>
</floats-group>
</pmc>
</record>
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