Pro-Syrian government fighters opened fire on a convoy as it prepared to leave rebel-held eastern Aleppo on Thursday, killing at least one person, a spokesman for the civil defence rescue service in the area told Reuters.
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Rebels are leaving the last enclave they hold in the city after major advances by President Bashar Assad's forces.
"[Pro-government fighters] fired at us and at ambulance vehicles and those people opening up the road," the spokesman said.
Earlier, the head of the ambulance service in the district, Ahmed Sweid, told pro-opposition Orient TV that three people had been injured in the incident.
"The convoy was shot at by regime forces and we have three injured, one of them from civil defence ... They were brought back to besieged areas," he told Orient TV.
An official with an Aleppo rebel group said the first convoy had reached the Ramousah junction on the way out when they came under fire. Rebel officials said they did not complete the crossing.
In a video interview posted to journalists via an online messaging service, a man who said he was a civil defence worker said snipers had fired on people as they tried to open the road for the ambulances to pass a government checkpoint out of the rebel-held sector.
"The ambulances were on the way to the crossing which was specified to us to for evacuating people and the regime forces started to shoot at us ... Even the men who tried to open the road with their trucks, they fired at them," the man in the video said.
Meanwhile, the powerful Syrian rebel group Ahrar al Sham said the agreement to evacuate civilians and rebels from Aleppo came after negotiators overcame what they said was obstruction by Iran and its militias to prevent the deal.
"There were Iranian efforts to exploit the situation in Aleppo and prevent any evacuation of our people from besieged Aleppo but in the end a deal was reached despite the Iranian intransigence," Ahmad Qura Ali, the spokesman for the group, told Reuters.
The convoy began a journey which will take patients through government territory into the rebel-held western Aleppo countryside as agreed in an evacuation deal this week, the war monitor said.
The Observatory added that no group of civilians or fighters had yet left.
The evacuation of rebel-held areas of Aleppo is back on track despite clashes overnight. Thursday morning, Syrian opposition groups and a military media unit run by the government's ally Hezbollah said evacuation is expected to begin "within hours."
A Syrian official source told Reuters early on Thursday that the operation to organize the departure of fighters from east Aleppo had begun and the International Committee of the Red Cross said it had been asked to assist with the evacuation. In a tweet, the ICRC said the operation would evacuate around 200 wounded people from rebel-held areas.
Overnight contacts between the parties succeeded in reviving a ceasefire that had originally come into effect late on Tuesday before breaking down, Hezbollah's media unit said.
An official from the Jabha Shamiya rebel group said the new truce came into effect at 2:30 A.M. local time on Thursday and a Reuters reporter in Aleppo said no fighting had been heard in the city since the early hours of the morning.
Such an exodus would end years of fighting for the city and mark a major victory for Syrian President Bashar Assad. An initial deal stalled on Wednesday, the planned evacuation failed to materialise and renewed fighting raged in the city.
Iran, one of Assad's main backers, had imposed new conditions, saying it wanted the simultaneous evacuation of wounded from two villages besieged by rebels, according to rebel and UN sources.
The evacuation plan was the culmination of two weeks of rapid advances by the Syrian army and its allies that drove insurgents back into an ever-smaller pocket of the city under intense air strikes and artillery fire.
By taking control of Aleppo, Assad has proved the power of his military coalition, aided by Russia's air force and an array of Shi'ite militias from across the region.
Rebels have been supported by the United States, Turkey and Gulf monarchies, but that support has fallen far short of the direct military backing given to Assad by Russia and Iran.
Russia's decision to deploy its air force to Syria 18 months ago turned the war in Assad's favour after rebel advances across western Syria. In addition to Aleppo, he has won back insurgent strongholds near Damascus this year.
The government and its allies have focused the bulk of their firepower on fighting rebels in western Syria rather than Islamic State, which this week managed to take back the ancient city of Palmyra, once again illustrating the challenge Assad faces reestablishing control over all Syria.