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Your obvious choice is Brno: http://www.jeremytaylor.eu/photos/brno-in-the-czech-republic/ Worth a day of your time.3
another vote for Brno.I was there in late May this year and found it a charming place to be, very friendly ,plenty to do and not over run with tourists.i was in the castle area and had the whole area to myself for the best part of a morning. The Dam area about 30 minutes by tram from the city center is a lovely peaceful place to spend a bit of time if the weather is nice.Plenty good places for meals and drinks,Pegas bar for beer made on the premisis and they serve the BEST wheat Beer I have ever tasted.4
I'd suggest Brno as well, it's too cold in January for sightseeing in smaller towns in the countryside, you need a place with plenty of indoor attractions and a good selection of restaurants and pubs. Brno can offer all these things as well as being about half way along the main rail line from Prague to Vienna. I second the recommendation for the Pegas pub and its fine beer, I always stop in there when I'm in Brno.Wildroo
The Goulash Train Blog
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Rob-Brno as mentioned above is a nice stop.
Not 100% sure it is on the way, but the town of Kutna Hora with the bone church is, in my opinion, a must see and I believe 80km or so outside Prague.
The Bone Church of Kuta Hora
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Just for the record Funspirit, no, the direct Prague - Vienna trains don't go through Kutná Hora, they follow the Prague - Brno main line through Pardubice. Some Prague - Brno trains do take the southern route through Kutná Hora, but none of the international trains do.Wildroo
The Goulash Train Blog

