要生成带有MD5检查的预签名URL，您可以使用以下Python代码示例：

import boto3
import hashlib
from botocore.client import Config

def generate_presigned_url_with_md5(bucket, key, expires_in):
    s3_client = boto3.client('s3', config=Config(signature_version='s3v4'))
    
    # 生成预签名URL
    presigned_url = s3_client.generate_presigned_url(
        ClientMethod='get_object',
        Params={
            'Bucket': bucket,
            'Key': key,
            'ResponseContentDisposition': 'attachment; filename={}'.format(key),
            'ResponseContentType': 'application/octet-stream',
            'ResponseContentLanguage': 'en',
            'ResponseExpires': expires_in
        }
    )
    
    # 计算MD5哈希值
    response = s3_client.get_object(
        Bucket=bucket,
        Key=key
    )
    object_data = response['Body'].read()
    md5_hash = hashlib.md5(object_data).hexdigest()
    
    # 将MD5哈希值添加到URL的查询参数中
    presigned_url_with_md5 = presigned_url + '&md5=' + md5_hash
    
    return presigned_url_with_md5

# 示例用法
bucket = 'your-bucket-name'
key = 'your-object-key'
expires_in = 3600 # 预签名URL的有效期（秒）

presigned_url_with_md5 = generate_presigned_url_with_md5(bucket, key, expires_in)
print(presigned_url_with_md5)

请注意，您需要将上述代码中的your-bucket-name和your-object-key替换为实际的S3存储桶名称和对象键。此外，您还可以根据需要调整其他参数，如ResponseContentDisposition、ResponseContentType等。

生成的预签名URL将包含一个名为md5的查询参数，其值为文件的MD5哈希值。您可以在验证URL时使用该MD5哈希值进行检查。