Objective To investigate efficiency and distribution of gene delivery to the injured tendons by microinjeetion and tissue reactions caused by different vectors. Methods By using a mi-croinjection technique, 10μl of pCMV-EGFP, pCAGGS-EGFP, AAV2-EGFP and Ad5-EGFP harboring enhanced green fluorescence protein (EGFP) gene were respectively injected to two sites of the proximal stump of 48 transected digital flexor tendons in 18 chickens. At days 3, 7, 14 and 21, the tendons wereharvested for observing distribution and expression of EGFP under a fluorescence microscope by using fro-zen tissue sections. The tendon sections were also stained with hematoxylin and eosin to examine inflam-mation caused by these vectors. The other 24 normal flexor tendons were served as controls. ResultsCompared with normal tendon tissues, the EGFP expression was observed in tendons at days 3, 7, 14 and21 after injection. The EGFP expression was observed at day 3 and reached peak at day 7 for all vectors.The EGFP expression was decreased at day 14 but seldom seen at day 21. EGFP was distributed evenly inthe injected tendon segment adjacent to the cut level. The EGFP expression in the tendons injected withAAV2-EGFP and Ad5-EGFP was higher than that with pCMV-EGFP and pCAGGS-EGFP injection, withinsignificant statistical difference upon the EGFP expression between AAV2-EGFP and Ads-EGFP vec-tors. Tissue reactions of the tendons caused by the liposome-plasmid vector ( including pCMV-EGFP and pCAGGS-EGFP) were the most prominent among all vectors. Infiltration of inflammatory cells, chiefly lymphocytes and neutrophilic granulocytes, were found. The tendons injected with AAV2 vectors presen-ted gentle inflammatory reactions. Conclusions Mieroinjection to two sites of each tendon stump deliv-ers the transgene to the entire tendon segment adjacent to the cut. Gene delivery by the AAV2 and Ad5 vectors has the highest cfficiency among four vectors tested, when expression level peaks at day 7 after in-jection and AAV2 vector causes the slightest tissue reactions in the tendons, indicating that the AAV2 vector is a promising gene delivery vector and microinjection is practical for tendon gene therapy.

Objective To investigate the relationship between the composition of vaginal microbiota and the course of cervical precancerous lesion.Methods A total of 64 vaginal swabs were collected from 22 healthy women, 18 CINⅠ patients and 24 CINⅡ/Ⅲ patients who visited Obstetrics and Gynecology of the Second Xiangya Hospital of Central South University during July 2014 and July 2015.The Bacterial genomic DNA was extracted and the V3 and V4 hypervariable regions of 16S rRNA were amplified and high-throughput sequenced.The abundance and composition of vaginal microbiota were analyzed by Uparse, Mothur and LefSe statistical software.Results There was no significant difference in Alpha diversity index between CINⅡ/Ⅲ group(Chao:63±32;ACE:72±38;Simpson:0.70±0.27;Shannon:0.70±0.63) and control group ( Chao:48±24;ACE:54±25;Simpson:0.71±0.27;Shannon:0.65±0.58)(W=192,P=0.11;W=189,P=0.10;W=281,P=0.72;W=241,P=0.62).The ACE(85±37) and Chao(66±25) values of CINⅠgroup were significantly different from those of the control group (ACE:54±25;Chao:48±24)(W=99,P=0.006;W=113,P=0.02).At the phylum level, 78.69%(309 020/392 722) of the vaginal microbiota in the control group was Firmicutes, 16%(62 846/392 722) was Actinobacteria.Firmicutes was reduced to 64.86%(208 422/321 318) and Actinobacteria increased to 27.71%(89 040/321 318) in CINⅠgroup.The composition of vaginal microbiotain in CINⅡ/Ⅲ group was similar to those of control group.At the genus level, the composition of vaginal microbiota were similar between CINⅡ/Ⅲ group and control group, with Lactobacillus as predominant genus[71.81%(307 658/418 424)], Gardnerella[12.91%(55 299/428 424)], others such as Prevotella, atopobium were less.In the CINⅠ group, the abundance of Lactobacillus was decreased to 56.26%(180 787/321 318), Gardnerella was increased to 19.62%(63 057/321 318), and Listeria was increased to 7.7%(24 746/321 318).The composition of vaginal microbiota in the most samples was classified as CSTⅢ and CSTⅠ, with Lactobacillus inersand and Lactobacillus crispatus were dominant respectively.There was no significant difference in the composition of vaginal microbiota between the three groups(χ2=2.72, P=0.949).LEfSe analysis showed that the abundance of bacteria in CIN group and control group were varied.At the genus level, there were significant differences in the abundance of Geobacter, Atopobium and Ureaplasma (P<0.05, P<0.05, P<0.01, respectively).At the species level, there was significant difference in the abundance of Ureaplasma urealyticum serotype 9 (P<0.01).Conclusion The diversity and the composition of vaginal microbiota were similar between CIN patients and healthy women, but the abundances of some bacteria were varied, with Ureaplasma increased in patients with CIN.