Objective To construct human secreted apoptosis-related protein 1 (SARP1) gene yeast two-hybrid bait vector so as to study the biological functions of the SARP1 gene in the scar tissue. Methods The target gene from SARP1 gene full-length DNA segment was amplified by PCR, the upstream and downstream primers of the SARP1 gene with restriction enzymes Nde I and Sal I were designed. pGBKT7-SARP1 recombination plasmid was constructed by ligating the vector and the PCR production and identified by PCR and sequencing. Further more, pGBKT7-SARP1 was transformed into competent AH109 which contained kanamycin for selecting positive clones and screened the positive clony on the plate of SD/-Trp. The toxicity and transcriptional activation were tested by the phenotype assay. Results SARP1 was amplified and cloned into pGBKT7 successfully, SARP1 gene sequence in recombinant plasmid pGBKT7-SARP1 was verified by gel electrophoresis and DNA sequencing analysis. The sequence of inserted SARP1 gene was the same as the corresponding sequence found in GenBank database. The recombinant pGBKT7-SARP1 plasmids and empty pGBKT7 vector could form white colonies on SD/-Trp plates and none could survive on SD/-Leu plates. Conclusion The recombinant pGBKT7-SARP1 gene yeast two-hybrid bait vector is successfully constructed.
Objective To explore the relationship between obstructive sleep apnea hypopnea syndrome ( OSAHS) and airway hyperresponsiveness ( AHR) . Methods 197 subjects suspected for OSAHS were enrolled in the study. They were all performed overnight polysomnogram ( PSG) monitoring and lung function test. Acoording to the results of FEV1% pred, they were performed bronchial provocation test( BPT)or brochial dilation test( BDT) . The relation between apnea hypopnea index ( AHI) and the degree of airway hyperresponsiveness ( AHR, expressed as PD20 -FEV1 ) was evaluated by linear correlation analysis. Results 117 patients were diagnosed as OSAHS, in which 28 cases were complicated with AHR( 3 cases with positive BDT result, 25 cases with AHR) . In 80 non-OSAHS patients, 7 cases were complicated with AHR. Theincidence of AHR was higher in the OSAHS patients compared with the non-OSAHS patients( 23. 9% vs 8. 8% , P lt; 0. 01 ) . AHI of OSAHS patients with AHR was higher than OSAHS patients without AHR[ ( 30. 3 ±5. 1) /h vs ( 23. 7 ±2. 4) /h, P lt;0. 01] . There was a positive correlation between AHI and degree of AHR in OSAHS patients with AHR( r=0. 62, P lt;0. 05, n=25) . Conclusion OSAHS is associated with an increased risk of AHR.
【摘要】 目的 原肌球蛋白是肌球蛋白主要相关蛋白之一，在细胞骨架与运动中起着重要的作用。探讨原肌球蛋白在增生性瘢痕中的作用，有助于揭示瘢痕挛缩的产生机制。 方法 收集2006年3月-2008年7月48例患者不同时期增生与非增生瘢痕组织标本，利用基因芯片筛选出的瘢痕相关基因——原肌球蛋白基因特异片段，制备成寡核苷酸探针与瘢痕组织切片进行原位杂交。同时，将各标本进行原代细胞培养，制备成纤维细胞爬片，进行原位杂交。 结果 原肌球蛋白基因在3、6个月的增生性瘢痕中表达均明显强于9、12个月增生性瘢痕及非增生性瘢痕，阳性细胞比例也高于9、12个月增生性瘢痕及非增生性瘢痕。 结论 瘢痕增生挛缩与细胞骨架运动的相关基因存在密切关系，而原肌球蛋白可能在其中起着重要作用。【Abstract】 Objective Tropomyosin is one of the proteins of cytoskeleton and cell movement. The aim of this study is to investigate the effect of gene expression of fibroblast tropomyosin on the formation and contraction of hypertrophic scar. Methods According to the results of differently expressed genes, in hypertrophic scar by gene microarray, topomyosin, one of the most important genes, was selected and made into oligonucleotide probe. Twenty-four hypertrophic scars and 24 non-hypertrophic scars and 12 normal skins were used and these scar were taken on 3, 6, 9, and 12 months after burned between March 2006 and July 2008. Frozen section and cultured fibroblasts were made to detect the expression of the gene by in situ hybridization. Results Expression of tropomyosin was defected in scar tissue, but those in the hypertrophic scar on 3 and 6 months after burn were significantly ber than those in the hypertrophic scar on 9 and 12 months after burn and non-hypertrophic scar. Conclusion Overexpression of cytoskeletal relative genes causes the contraction of scar and tropomyosin acts leading and key functions.
In recent years, pathogenic metagenomic next-generation sequencing (mNGS) technology has become more and more widely used in the field of clinical infection. Driven by clinical needs, mNGS technology is constantly being optimized and developed. From manual operation to automated process, from simple qualitative detection to quantitative monitoring, from infection identification to multi-dimensional diagnosis of “infection+tumor”, certain research results have been achieved. Of course, there are still some limitations in clinical application of this technology. For example, there are many mNGS detection related reagents in China, but a systematic and complete quality management control and evaluation system has not been established. Further research is needed. This article summarizes the development and application of pathogen metagenomic sequencing technology in recent years, as well as the standardization and normalized process of mNGS detection, in order to provide a reference for pathogen mNGS sequencing to better assist clinical infection diagnosis.
Objective To explore the clinical characteristics of patients with severe pandemic H1N1 Influenza in Sichuan and risk factors related to patients’ prognosis. Methods We observed 135 severe patients who came to hospitals for pandemic H1N1 Influenza from 12 cities in Sichuan, China,between September 12, 2009 to December 14, 2009, and described their baseline characteristics, treatment,and outcomes. A stepwise multiple Logistic-regression analysis was used to evaluate the independentpredictors of death. Results Of the 135 patients we studied, 86 patients were male. The average age was ( 28. 2 ±19. 3) years old, while patients between 19 to 45 years of age accounted for 47. 4% . 96 patients ( 71. 1% ) presented with fever. 51 patients( 37. 8% ) had comorbid conditions. The most frequent organdysfunction was seen in lung ( 71. 1% ) , liver( 27. 4% ) and cardia( 24. 4%) ; 130 patients( 96. 3% ) had received oseltamivir, 26 patients ( 19. 3% ) required mechanical ventilation. 12 of the 135 patients died.Compared with the survivors, patients who died were more likely to have a higher age, lower average bloodpressure when admitted, more organ dysfunction, and more likely to have cardia or nervous system dysfunction. The nonsurvivors also seemed to have less opportunity to be exposed to neuraminidase inhibitors, and have more demand for mechanical ventilation. The P value were all under 0. 05. The multipleLogistic-regression analysis showed the independent predictors of death were the average blood pressure when admitted and the demand for mechanical ventilation . The P value were both under 0. 05. The OR value was 0. 86(95% CI 0. 002-0. 936) and 13. 86( 95% CI 1. 146-16. 583) , respectively. Conclusions For these severe patients with pandemic H1N1 Influenza we study, the male patients are more than female. Most patients are between 19 to 45 years of age. The most frequent organ dysfunction is seen in lung, liver and cardia. The mortality of these patients is 8. 9% . Compared with the survivors, patients who died were morelikely to have a higher age, lower average blood pressure when admitted, more organ dysfunction, and more likely to have cardia ornervous systemdysfunction. The nonsurvivors also seemed to have less opportunity to be exposed to neuraminidase inhibitors, and more demand for mechanical ventilation. The multiple Logisticregression analysis showed the independent predictors of death are the average blood pressure and the demand for mechanical ventilation. The OR value is 0. 86 ( 95% CI 0. 002-0. 936) and 13. 86 ( 95% CI1. 146-16. 583) respectively.