不要以这种方式遍历文件。而是使用for循环。
for line in f:
vowel += sum(ch.isvowel() for ch in line)
实际上，您的整个程序就是：
VOWELS = {'A','E','I','O','U','a','e','i','o','u'}
# I'm assuming this is what isvowel checks, unless you're doing something
# fancy to check if 'y' is a vowel
with open('filename.txt') as f:
vowel = sum(ch in VOWELS for line in f for ch in line.strip())
就是说，如果while由于某些误导的原因您真的想继续使用循环：
while True:
line = f.readline().strip()
if line == '':
# either end of file or just a blank line.....
# we'll assume EOF, because we don't have a choice with the while loop!
break
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