[Clinical application and progress of airway stent in subglottic stenosis].

Laryngotracheal stenosis is caused by various reasons of laryngotracheal cartilage stent malformation, collapse or defect, laryngotracheal mucosa scar formation or submucosal tissue hyperplasia, eventually resulting in dyspnea. Subglottic stenosis refers to the airway stenosis from subglottic to the lower margin of the cricoid cartilage, which is a special type of laryngotracheal stenosis. The most common cause is iatrogenic injury, such as prolonged tracheal intubation and tracheotomy. Currently, the main treatments include surgical treatment, tracheostomy, endoscope-guided stent implantation and drug therapy. As for the patients who have dyspnea not suitable for surgery or in urgent need of preoperative transitional treatment, stent implantation guided by respiratory endoscopy has become an important treatment. In this paper, we reviewed 51 literatures on stent implantation of subglottic stenosis since 1994 retrieved from PubMed, CBM, CNIT, Wan-fang and VIP databases, focusing on the comparison of the efficacy, complications and prognosis of metal stent, hourglass-shaped DUMON stent, straight-type DUMON stent and Montgomery T tube and investigated the clinical application of endoscope-guided stent implantation in subglottic airway stenosis. Literature studies have shown that compared with DUMON silicone stents and metal stents, T tube has more significant advantages, higher treatment success rate and lower complication rate. Therefore, Montgomery T tube is more suitable for long-term treatment of patients with subglottic stenosis. With the improvement and update of new technology and materials, the vigorous development of new airway stents also provides a new stent treatment mode with better histocompatibility, fewer complications and customized options for the patients with subglottic stenosis.

The Study of Hotspots on Nursing Dissertation Based on Co-Word Analysis in China.

All the doctoral dissertations and master's theses of "nursing" in CNKI database were retrieved to explore the hot spots on nursing research field in China, we processed, visualized the data above, and analyzed those data by using co-word analysis informetrics method. Then, the figure for co-occurrence analysis of high-frequency keywords in dissertation on nursing research were plotted, which represented nine topics and could help health staff to understand hot topics in nursing research field.

The clinicopathological significance of REIC expression in colorectal carcinomas.

REIC is down-regulated in immortalized cell lines compared with the parental normal counterparts, and could inhibit colony formation, tumor growth and induce apoptosis. Here, its expression was examined by immunohistochemistry on tissue microarray containing colorectal non-neoplastic mucosa (NNM), adenoma and adenocarcinoma. Colorectal carcinoma tissue and cell lines were studied for REIC expression or its secretory level by Western blot, RT-PCR or enzyme-linked immunosorbent assay (ELISA). The results demonstrated that REIC was differentially expressed in Colo201, Colo205, DLD-1, HCT-15, HCT-116, HT-29, KM-12, SW480, SW620, and WiDr with its secretion concentration less than 300 pg/mL. Carcinomas showed statistically lower REIC expression than matched NNM with no difference for protein content. Immunohistochemically, REIC expression was significantly decreased from NNM, adenoma to adenocarcinoma (p<0.05). REIC expression was negatively correlated with depth of invasion, TNM staging, dedifferentiation, Capase-3 and nuclear inhibitor of growth 5 (ING5) expression (p<0.05), while not with age, sex, tumor size, lymphatic or venous invasion, or lymph node metastasis (p>0.05). Kaplan-Meier analysis indicated that REIC expression was not associated with the prognosis of colorectal carcinomas (p>0.05). Cox's analysis demonstrated that lymphatic and venous invasion, lymph node metastasis, and UICC staging were independent prognostic factors for carcinoma (p<0.05). Our study indicated that down-regulated REIC expression might play an important role in colorectal adenoma-adenocarcinoma sequence and subsequent progression. Aberrant REIC expression might be employed as a good marker of pathogenesis and development of colorectal carcinomas.

[Splicing and stability of intron in the expression retroviral vector with human clotting factor IX].

To study the role of intron in the expression of hFIX, retroviral vectors with intron containing hFIX were constructed. It is fundamental for the intron study whether the intron constructed in retroviral vector can be steadily transferred into target cell. First, we constructed two forward-orientation retroviral vectors: G1NaC-i-IX contains the exogenous intron from IL-2, and G1NaC-i'-IX contains the truncated intron I from hFIX gene, covering the splicing donor and acceptor sequences. RT-PCR result indicated that intron in the forward-orientation retroviral vector was spliced after packaging in PA317. Then, reverse-orientation retroviral vectors G1NaC-i'-IXR and G1NaPAIXi' BAM were constructed, in which the reverse and complimentary sequences of hFIX gene with intron appeared in retroviral RNA. RT-PCR assay combined with ELISA test indicated that intron was retained after packaging and hFIX gene with intron constructed in the reverse-orientation retroviral vector can be transduced intact and expressed hFIX at a high level in vitro.

Increment of hFIX expression with endogenous intron 1 in vitro.

This paper probes into the feasibility of increasing expression level of hFIX gene with endogenous intron 1 sequence. hFIX minigene was obtained with middle sequence truncated intron 1 inserted into the relative site of hFIX cDNA, and plasmid vector pKG5i'IX, retroviral vector GINaCi'IX were constructed. These vectors were transduced into target cells of PA317, C2C12, primary rabbit skin fibroblasts (RSF) and primary human skin fibroblasts (HSF). The expression level of mixed colonies are PA317/pKGoi'IX, 151 ng/10(6) cells/24h; PA317/G1NaCi'IX, 308 ng/10(6) cells/24 h; C2C12/G1 NaCi'IX, 188 ng/10(5) cells/24 h; RSF/G1NaCi'IX, 1929 ng/10(5) cells/24 h; HSF/G1NaCi'IX, 1646 ng/10(6) cells/24 h. These results indicated that hFIX minigene with intron 1 is able to increase the expression level to about 3 times of that of hFIX cDNA. Meanwhile, in order to study the application of hFIX minigene in the retroviral-mediated gene transfer system and refrain from intron splicing during viral production, a retroviral vector G1NaCi'IXR with reversely inserted hFIX minigene expression cassette was constructed. The expression level of reverse constructor in PA317 cells was 390 ng/10(6) cells/24 h with 79% of bioactivity. PCR detection of HT/G1NaCi'IXR cells infected with PA317/G1NaCi'IXR supernatant confirmed the existence of intron 1 sequence. These results suggested that expression vector with forward-inserted intron1-carrying hFIX expression cassette can be used in directed gene transfer, but when using the retroviral-mediated gene transfer system, reversely-inserted intronl-carrying hFIX expression cassette should be considered.