10.3969/j.issn.2095-4344.2013.23.018

In order to set up a base for stem cells to be widely used in clinical medicine, we tried to optimize, in this study, the technique that induces human mesenchymal stem cells (hMSCs) to differentiate into neural stem cells by using cerebrospinal fluid (CSF) from the different groups. After the induction, presence of neural stem cells was confirmed with microscope observation, flow cytometry analysis, immunohistochemistry and fluorescent immunohistochemistry. At the same time, we also compared and analysed the data of the number of stem cells when it totally met the requirements for clinical treatment and the days required. At last, we confirmed that hMSCs could be induced to differentiate into neural stem cells, and that the number of cells totally met the requirements for clinical treatment. But there were some differences both in the number of cells and the days required. Among the groups, the group that marrow mesenchymal stem cells from patients own induced by CSF from healthy volunteers used the shortest time and the quantity of the cells was significantly higher than those of the others.
Cell Differentiation ; Cerebrospinal Fluid ; chemistry ; Culture Media ; chemistry ; Flow Cytometry ; Humans ; Immunohistochemistry ; Mesenchymal Stromal Cells ; cytology ; Neural Stem Cells ; cytology

Child, Preschool ; China ; epidemiology ; Enterovirus A, Human ; genetics ; Female ; Hand, Foot and Mouth Disease ; epidemiology ; Humans ; Infant ; Male ; Molecular Epidemiology

OBJECTIVETo identify the enterovirus from stool samples of patients with hand, foot and mouth disease(HFMD) in Guangzhou from 2010 to 2012 and to perform phylogenetic analysis of the VP1 gene sequences of coxsackievirus A4 and coxsackievirus A10.

METHODSA total of 5 484 samples of suspected cases of HFMD which Guangzhou Center for Disease Control received from 2010 to 2012 were collected.Virus RNA was tested by nested RT-PCR method as human enterovirus 71, coxsackievirus A16, coxsackievirus A4, coxsackievirus A10 and other enteroviruses positive, and 4 111 samples were positive. Phylogenetic tree was constructed by partial VP1 gene sequences of coxsackievirus A4 and coxsackievirus A10 to perform phylogenetic analysis.

RESULTSIn 4 111 enterovirus-positive samples, the positive rate of EV71, CoxA16, CoxA10 and CoxA4 was 35.1% (1 443/4 111) , 30.7% (1 261/4 111) , 2.0% (82/4 111),0.8% (31/4 111) respectively. Different enterovirus-positive rate was statistically significant (χ(2) = 148.34, P < 0.05) .Incidences of coxsackievirus A4 positive was highest in 3-year old children as 1.3% (7/534) , and that of coxsackievirus A10 positive was highest in 0-year old children as 3.7% (34/914) . The highest positive rate of diagnosed coxsackievirus A4 positive cases was admitted in April(2.6%, 12/460) , and the highest positive rate of diagnosed coxsackievirus A10 positive cases was admitted in August 4.3% (12/278). Phylogenetic analysis indicated that all the CoxA4 stains were divided into subtype A and subtype B, and the CoxA10 stains were divided into subtypes A, subtype B and subtype C. The VP1 gene nucleotide sequences of CoxA4 and CoxA10 this study measured both belonged to subtype A.

CONCLUSIONSThe VP1 gene nucleotide sequences of CoxA4 and CoxA10 in Guangzhou from 2010 to 2012 both belonged to subtype A.

Child ; China ; epidemiology ; Enterovirus A, Human ; Hand, Foot and Mouth Disease ; Humans ; Molecular Epidemiology ; Phylogeny ; Polymerase Chain Reaction ; RNA, Viral

OBJECTIVETo identify the pathogen and characteristics on a case of hand-foot-mouth disease (HFMD) caused by coxsackie-virus A6 (CA6) associated with vaccine-derived poliovirus (VDPV) co-infection.

METHODSField epidemiological study at the epidemic area was conducted and 16 stool samples including from the patient and close contacts were collected for isolation and identification of the enterovirus (EV). 21 stool samples from patients diagnosed as HFMD were collected in the same hospital at the same month to detect CA16,EV71, CA6 and PV by real-time RT-PCR or RT-PCR. The VP1 gene of the CA6 was amplified by RT-PCR and PCR products were sequenced and analyzed.

RESULTSThe patient showed only HFMD symptoms, but no symptoms related to acute flaccid paralysis (AFP). No EVs were isolated from 16 samples collected from the patient and close contacts. And no AFP cases were found by an active search. A total of 21 samples from patients diagnosed as HFMD were collected in the same hospital at the same month and 4 were found to be EV71, 2 were CA16 and 15 (include the patient)were CA6. Only this patient was found to have had VDPV II infection. The CA6 VP1 gene was amplified from the HFMD patient and 9 other cases from the same hospital at the same month. Nucleotide sequences of the VP1 gene among the 9 strains shared 98.9%-100.0% in homology and 96.0%-100.0% in the deduced amino acid sequences. Phylogenetic analysis of the VP1 sequences categorized the 9 strains into the same branch. There were 6 nucleotides changes including U2909A between the VP1 region of the VDPV strain of the case and Sabin II. Results from phylogenetic analysis on the VP1 sequences indicated that the VDPV strain of the case was different from other VDPVs strains isolated in the world.

CONCLUSIONThis case was a HFMD which caused by CA6 co-infection with VDPV II and the VDPV was newly discovered. HFMD symptoms of the case were caused by CA6. The reason why this case did not have AFP symptoms was probably due the protective effect of IPV vaccine. No AFP cases were found by the active search for AFP cases conducted in the area, which indicated that VDPV did not cause virus circulation in this area.

Child, Preschool ; Coinfection ; Enterovirus A, Human ; isolation & purification ; Female ; Hand, Foot and Mouth Disease ; complications ; virology ; Humans ; Poliovirus Vaccines ; adverse effects

Objective: To find out the source and the epidemic pattern of norovirus outbreak in July, 2016 to June, 2017 in Guangzhou. Methods: The stool samples and clinical information of diarrhea cases were collected by the sentinel hospitals and CDCs; a real-time RT-PCR method was used to detect the norovirus nucleic acids from the samples, the positive ones were amplified and sequenced; the partial sequences of norovirus were aligned by an online BLAST alignment, and a phylogenetic tree was constructed by a neighbor-joining method . Results: A total of 854 cases with infectious diarrhea were reported by Guangzhou diarrhea surveillance network from July, 2016 to June, 2017; the gender ratio (male versus female) was 1∶0.67; 78.33% of the cases were preschool children under the age of 7 years. Totally 220 samples were detected norovirus G II+ (25.76%, including 5 double-positive samples with G I+ ). GII.Pe-GII.4.Sydney_2012 was the prevalent genotype in the second half of 2016 (94.64%), which was replaced by GII.P16-GII.2 in the first half of 2017 (67.65%). Since September 2016, the reported number of norovirus-caused diarrhea epidemic was increased gradually; the peak of epidemic curve emerged in February to March of 2017, and the number started to decrease since April. In May to June there were only 2-3 epidemics reported monthly. All the endemics from September to November 2016 were caused by genotype GII.Pe-GII.4.Sydney_2012; the endemics from December 2016 to April 2017 were mainly caused by genotype GII.P16-GII.2. Some samples from kitchen workers and babysitters were detected GII+ , which was consistent with the result of the cases′ samples. Conclusions It was the first time that the novel GII.P16-GII.2 recombinant strain outbroke occurred in Guangzhou City and homology analysis also suggested that GII.P16-GII.2 was the main source of those epidemics in 2016 -2017 winter and spring season. Furthermore, The kitchen workers and babysitters may have played an important role in the spread of norovirus.