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.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Non-Small-Cell Lung ; genetics ; DNA Mutational Analysis ; methods ; DNA, Neoplasm ; genetics ; Female ; Humans ; Lung Neoplasms ; genetics ; Male ; Middle Aged ; Mutation ; Mutation Rate ; Receptor, Epidermal Growth Factor ; genetics

Objective:Cytosolic 5'-nucleotidase (CN-Ⅱ), a nucleotide kinase, exhibits both 5'-nucleotidase and nucleoside phos-photransferase activities. Abnormal CN-Ⅱexpression may be correlated with the resistance of nucleoside analogs in anticancer drugs. This study was designed to investigate CN-Ⅱexpression in human non-small cell lung cancer (NSCLC) tissues and its correlation with the clinicopathological parameters as well as the prognosis of patients treated with gemcitabine. Methods:Immunohistochemistry was used to detect CN-Ⅱexpression in 116 cases of paraffin-embedded NSCLC samples. The correlations with the clinicopathological pa-rameters and the response to gemcitabine chemotherapy of CN-Ⅱwere analyzed through the Chi-square test. Log-rank test was used to determine whether or not CN-Ⅱexpression is correlated with the overall survival of patients. Results:The positive rate of CN-Ⅱwas 53.4% in 116 NSCLC tissues. No significant correlation existed between CN-Ⅱ expression and the clinicopathological parameters. Among the 67 of the 116 patients who received gemcitabine chemotherapy, those with tumor progression (positive rate of 57.6%) exhib-ited higher CN-Ⅱexpression than those with therapeutic efficacy (positive rate of 30.4%, P=0.008) and disease-control chemotherapy (positive rate of 36.7%, P=0.013). The progression-free survival was 4.5 and 5.5 months in the CN-Ⅱ-positive and CN-II-negative groups, respectively, with significant differences (95%CI:4.452 to 6.148, P=0.041). Correspondingly, the overall survival was 9.5 and 11.0 months in the two groups (95%CI:8.667 to 13.333, P=0.282). Conclusion:CN-Ⅱmay be a prognostic factor for gemcitabine chemotherapy in NSCLC patients.

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.

Objective: To study the prevalence and antimicrobial susceptibility of Ureaplasma urealyticum (Uu) and Mycoplasma hominis (Mh) in Changsha, and provide laboratory evidence for clinical drug use. Methods: A retrospective study was conducted to analyze 53 006 specimens of suspected genital mycoplasma infection in Xiangya Second Hospital of Changsha District and Hunan Wangwang Hospital from 2010 to 2017, and to analyze the infection rate and drug resistance rate of Uu and Mh. Results: From 2010 to 2017, a total of 53 006 specimens were detected, where there were 16 830 cases of Uu infection, the infection rate was 31.75%; 2 471 cases of Mh infection, the infection rate was 4.66%; and 1 071 cases of Uu and Mh mixed infection, the infection rate was 2.02%. Male Uu infection rate was 19.48%(5 989/30 749), which was lower than the female infection rate 48.71%(10 841/22 257) (χ²=5 091, P<0.001); male Mh infection rate was 3.16%(973/30 749), lower than female infection rate 6.73%(1 498/22 257) (χ²=369,P<0.001). The population of genital mycoplasma infection is concentrated between 20 and 40 years old, accounting for 71.76% (12 077/16 830). The drug resistance rates of Uu and Mh to doxycycline and minocycline were less than 2%, while the drug resistance rate to quinolones was higher; The resistance rate of Uu to macrolide antibiotics such as erythromycin, josamycin and clarithromycin were less than 2%, while the resistance rate to azithromycin, erythromycin and roxithromycin were higher, 29.74%(5 006/16 830) and 53.74%(9 045/16 830), respectively, and the resistance rate of Mh to macrolide antibiotics (except josamycin) was higher than 90%.Between 2010 and 2017, a gradually increasing resistance of ureaplasmas to azithromycin, from 3.81% (46/1 206) in 2010 to 53.15% (1 503/2 828) in 2017, and decreasing resistance to gatifloxacin and thiamphenicol were observed, from 76.78% (926/1 206) and 60.28% (727/1 206) in 2010 decreased to 34.23% (968/2 828) and 37.87% (1 071/2 828) in 2017, respectively. The resistance rate of Mh to gatifloxacin and thiamphenicol were decreased, from 68.93% (122/177) and 41.81% (74/177) in 2010 to 53.54% (159/297) and 21.21% (63/297) in 2017, respectively. Conclusions Doxycycline, minocyclinum and josamycin are good treatment options for genital mycoplasma in Changsha. The resistance rate of Uu to azithromycin is increasing, suggesting that the abuse of azithromycin is present in Changsha, and indicating that better management of antibiotics is necessary.

Immune checkpoint inhibitors (ICI) have transformed the treatment landscape of advanced non-small cell lung cancer (NSCLC). Biomarkers are essential for guiding precision immunotherapy. Tissue-based programmed death ligand 1 (PD-L1) expression and tumor mutational burden (TMB) are currently widely used biomarkers for selecting patients for immunotherapy. However, tissue specimens are often difficult to reach and couldn't overcome spatial and temporal heterogeneity. Blood biomarkers offer an alternative non-invasive solution that could provide a complete insight on patient's immune status and tumor as well, and show their potential in predicting the outcome as well as in monitoring response to immunotherapy. In this article, we summarize current knowledge on blood biomarkers in NSCLC patients treated with ICI, and we hope to provide more references for development of novel biomarkers.
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Bone is one of the most metastatic sites of advanced malignant tumors. With the continuous improvement of diagnosis and treatment of malignant tumors, the survival time of patients is prolonged and incidence of bone metastases also increases. Lung cancer is the leading cause of cancer-related mortality worldwide. It is estimated that the incidence of bone metastases in patients advanced lung cancer is about 30%-40%. The traditional diagnosis of bone metastases in lung cancer is based on clinical symptoms, X ray, computed tomography (CT), magnetic resonance imaging (MRI) and pathology. Recently, a large number of exploratory studies have reported blood biomarkers as indicators of bone metastasis screening and efficacy evaluation. In this review, we summarize the progress of biomarkers in diagnosis of bone metastases of lung cancer.
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Biomarkers, Tumor ; metabolism ; Bone Neoplasms ; metabolism ; physiopathology ; secondary ; Humans ; Lung Neoplasms ; pathology ; Osteogenesis