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.

Background and objective Small-cell lung cancer (SCLC) is an aggressive disease for which the mainstay of treatment is cytotoxic chemotherapy. Despite good initial responses most patients will relapse or progress atfer the ifrst-line therapy. hTe evidence of a beneift from second-line chemotherapy is limited in patients with relapsed/advanced SCLC. Some drugs are recommended by guidelines, but more regimens are formulated based on experience in clinical. So we conducted this retrospective study in order to compare the effcacy and safety of different second-line treatment regimens. Methods We totally analyzed 309 patients received second-line treatment in our retrospective study. 157 patients received best supportive care (BSC), and the rest 152 patients received second-line chemotherapy. hTe Kaplan-Meier method survival curves and Log-rank test were used to analysis the differences among different groups. hTe endpoints were objective response rate (ORR), disease control rate (DCR), progression-free survival (PFS), and overall survival (OS). Results Patients administered second-line chemotherapy lived signiifcantly longer, with a total OS from ifrst-line therapy of 11.5 mo compared to 6.0 mo in patients with best supportive care alone (P<0.001), and the ORR, DCR, PFS and OS of the former (including the sensitive dis-ease and resistance/refractory disease patients) were obviously better than that of the latter. hTe ORR and DCR of the patients who received second-line chemotherapy is 39.5%and 59.2%, respectively. hTe median PFS and OS from second-line chemo-therapy were 3.3 mo and 5.3 mo. hTe patients who received second-line chemotherapy were divided by types of second-line regimens. hTe sensitive disease patients were from group A (VP-16-based rechallenge) and group B1 (CPT-11-based regimen). hTe ORR of the two groups were 48.6%and 35.3%, and the DCR were 68.6%and 58.8%, respectively. hTere was no statistically signiifcant difference (P=0.264;P=0.400). hTe median PFS from second-line chemotherapy of the two groups were 4.0 mo and 3.0 mo, and the second-line median OS were 6.5 mo and 4.5 mo. hTere was no statistic difference (P=0.432;P=0.508). hTe resistance/refractory disease patients were divided into group B2 (CPT-11-based regimen), group C (PTX/DXL-based regi-men) and group D (TPT-based regimen). hTere was no statistic difference in second-line ORR, DCR and median PFS among the three groups (P value is 0.521, 0.528 and 0.775, respectively);hTe median OS from second-line chemotherapy of the group D is longer than that of group B2 and group C, with statistical difference (P=0.043;P=0.030). hTe differences of grade III-IV hematologic toxicities among the four subgroups were not statistically different. hTe incidence of diarrhea in non-hematologic toxicities in patients who received irinotecan as second-line chemotherapy was higher than other three subgroups (P=0.029). Conclusion Patients who progressed atfer the completion of ifrst-line chemotherapy can gain survival beneift. hTe response and the PFS of the different second-line chemotherapies were similar. hTe patients who received the TPT-based regimen may gain longer overall survival than other resistance/refractory disease patients.