You don't actually need jQuery for this - straight vanila JS will do it.
In either case you use a selector to get the parent table (#Main) and then within that another selector to get the nested table and its tr.length;
The following snippet shows your table and a non jquery approach as well as a jquery aporach.
Also note that you should rejig your code, IMO. Its always better to separate out the CSS from the HTML - so all that inline CSS should be moved to the CSS block.
And you should, for semantically correct table structure - have your th's in a thead element and the rest in a tbody element.
const nonjQueryNestedTable = document.querySelector('#Main table');
const nonjQueryNestedTableTrLength = nonjQueryNestedTable.querySelectorAll('tr').length;
console.log('non-jquery: ' + nonjQueryNestedTableTrLength); // gives 3
$(document).ready(function(){
const nestedTable = $('#Main').find('table');
const nestedTableTrLength = nestedTable.find('tr').length;
console.log('jquery: ' + nestedTableTrLength); // gives 3
})
<script src="https://cdnjs.cloudflare.com/ajax/libs/jquery/3.3.1/jquery.min.js"></script>
<table id="Main">
<thead>
<tr>
<th style="width: 45px;"><input style="margin:0" type="checkbox" id ="CheckAllTable" class="checkAll" name="checkAll"></th>
<th style="width: 100px;">Column1</th>
<th style="width: 330px;">Column2</th>
<th style="width: 112px;">Column3</th>
</tr>
</thead>
<tbody id ="Tbody">
<tr>
<td><input style="margin:0" type="checkbox" name="check"></td>
<td><a target="_blank" href="https://www.example.com/">xxx testing</a></td>
<td>
<span>
<table style="border-collapse:collapse;table-layout:auto;width:100%;border:1px;">
<tr>
<th style="text-align:center;border:1px solid orange;padding:10px;width:20%;">subcolumn1</th>
<th style="text-align:center;border:1px solid orange;">subcolumn2</th>
<th style="text-align:center;border:1px solid orange;">subcolumn3</th>
<th style="text-align:center;border:1px solid orange;">subcolumn4</th>
</tr>
<tr>
<td style="text-align:left;border:1px solid orange;padding:10px;width:20%;">AAA</td>
<td style="text-align:left;border:1px solid orange;padding:10px;width:50%;">BBB</td>
<td style="text-align:left;border:1px solid orange;padding:10px;width:20%;">CCC,DDD</td>
<td style="text-align:left;border:1px solid orange;padding:10px;width:10%;">EEE</td>
</tr>
<tr>
<td style="text-align:left;border:1px solid orange;padding:10px;width:20%;">AAA1</td>
<td style="text-align:left;border:1px solid orange;padding:10px;width:50%;">BBB1</td>
<td style="text-align:left;border:1px solid orange;padding:10px;width:20%;">CCC1</td>
<td style="text-align:left;border:1px solid orange;padding:10px;width:10%;">EEE1 </td>
</tr>
</table>
</div>
</span>
</td>
<td><span>08/18/2020</span></td>
</tr>
</tbody>
</table>
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