Establishment of Surfactant-associated Protein A Suicide Gene System and Analysis of Its Activity

Summary： Alveolar epithelial type II （AT II） cells are essential for lung development and remodeling, as they are precursors for type I cells and also produce other non-repair cells （fibroblasts）. Progenitor ceils are believed to possess capability of multi-potent transdifferentiation, which is closely related to the niche, suggesting the importance of establishment of a lung progenitor cell niche model. We hypothesized that pulmonary surfactant-associated protein A （SPA） suicide gene system would cause AT II cell to kill itself through apoptosis and leave its niche. In vitro, the recombinant adeno-associated virus vectors-SPA-thymidine kinase （rAAV-SPA-TK） system was established to get targeted apoptotic AT II cells. The apoptosis of AT II cells was detected by using MTT. The results showed that cloned SPA gene promoter had specific transcriptional activity in SPA high expression cells, and SPA high expression cells （H441） transfected with TK gene had higher sensitivity to ganciclovir （GCV） than SPA low expression cells （A549）. In vivo, increased apoptosis of AT II cells induced by GCV in rAAV-SPA-TK system was observed by TUNEL. Finally, the successful packaging and application of rAAV-SPA-TK system provide experimental basis to get specific lung progenitor cell （AT II） niche in vitro and in vivo.

Surfactant Protein B 1580 Polymorphism Is Associated with Susceptibility to Chronic Obstructive Pulmonary Disease in Chinese Han Population

Summary: Whether surfactant protein B (SP-B)-18A/C and 1580C/T polymorphism were associated with susceptibility to chronic obstructive pulmonary disease (COPD) in Chinese Han population was investigated. After genomic DNA was isolated from blood of COPD smokers and control smokers, the genotypes of SP-B-18A/C and SP-B1580C/T polymorphism loci were determined by polymerase chain reaction-restriction fragment length polymorphism analysis (PCR-RFLP) respectively. The results showed that there was significant difference in genotypes distribution frequency of SP-B1580C/T polymorphism locus between COPD smokers and control smokers. C→T mutation rate (including TT homozygote and CT heterozygote) in COPD smokers was higher than in control smokers (57.9 % vs 41.7 %, χ2=4.93, P<0.05), whereas there was no significant difference in genotypes distribution frequency of SP-B1580-18A/C locus between COPD smokers and control smokers. The allele frequency (29.1 %) of SP-B1580-18A/C locus is lower than T allele (70.9 %) in Chinese Han Population, and the distribution was different from that in Mexican, in which, the A and T allele frequencies were 85 % and 15 % respectively. It was concluded that SP-B1580 T allele was probably associated with increased susceptibility to COPD in Chinese Han population; The polymorphism of SP-B-18A/C locus maybe varied with race.

Surfactant Protein A Polymorphism Is Associated with Susceptibility to Chronic Obstructive Pulmonary Disease (COPD) in Chinese Uighur Population

This study investigatedexamined the correlation between surfactant protein-A (SP-A) polymorphism and the susceptibility of cvhronic obstructive pulmonary disease (COPD) in Xinjiang Uighurs. Genomic DNA was extracted from peripheral blood of 194 COPD smokers and 201 healthy smokers of Uighur who were hospitalized in or paid a visit to one of the four Xingjiang-based hospi-tals involved in the study, betweenfrom March 2009 to December 2010. Single nucleotide polymor-phisms (SNPs) were studied on A/G atwithin amino acid aa62 (CCA/CCG rs1136451) and C/T within aa219 (CGG/TGG, rs4253527) in SP-A. Genotypes were determined by using the TaqMan polymerase chain reaction (PCR). Our results showed that genotype frequencies were different be-tween the COPD and normal smokers infor aa62 (χx2=6.852, P=0.033). There were also significant differences in allele genotype frequencies between the COPD and the control and allele G might de-crease the risk COPD (χx2=6.545, P=0.011; OR=0.663; 95% CI: 0.484–0.909). The result suggested We were led to conclude that polymorphism of aa62 (CCA/CCG, rs1136451) of SP-A may be asso-ciated with the susceptibility to COPD in Xingjiang Uighurs.

Discordantly Elevated Erythrocyte Sedimentation Rate (ESR) and Depressed C-Reactive Protein (CRP) Values in Early Diagnosis of Pulmonary Tuberculosis Patients in Maiduguri, Nigeria

Background: ESR and CRP measurements reflect different aspects of systemic inflammation. Generally, they are either elevated or depressed at the same time. This study was aimed at evaluating the clinical ability of these markers in the early diagnosis of pulmonary tuberculosis. Materials and Methods: A total of 50 (male 39, female 21) patients who tested positive to Acid Alcohol Fast Bacilli (AAFB) were studied. ESR and CRP values were estimated using standard procedures. Results: The mean ESR value among the AAFB positive patients was 53.16 ± 4.92 mm/hr while the CRP value was 0.273 ± 0.035 mg/L. Gender and age were not found to have any influence on the ESR and CRP values. No relationship was found to exist between ESR and CRP (r = 0.17;p = 0.235) and age (r = 0.125;p = 0.388). Conclusion: A moderately elevated ESR and low CRP values exist at the early diagnosis of tuberculosis. There is no correlation between ESR and CRP at the onset of tuberculosis;hence CRP cannot be used as a screening tool for early diagnosis of tuberculosis.

Expression of Heat Stress Protein 70 mRNA in Patients with Chronic Obstructive Pulmonary Disease and Its Significance

The effects of cigarette smoke extract (CSE) on the expression of heat stress protein 70 (Hsp70) in human bronchi smooth muscle cells were investigated in vitro, and the changes in Hsp70 mRNA in the patients with chronic obstructive pulmonary disease and their significance were explored. Human bronchi smooth muscle cells were cultured with CSE at the different concentrations. The expression of Hsp70 mRNA and Hsp70 was detected by reverse translation-polymerase chain reaction (RT-PCR) and Western blotting respectively. Levels of Hsp70 mRNA and Hsp70 in lymphocytes from 20 patients with COPD and 20 healthy smoking control subjects were measured by RT-PCR and Western blotting. The results showed the expression of both Hsp70 mRNA and Hsp70 was decreased conformably in human bronchi smooth muscle cells treated with CSE at certain concentration in vitro. The A values of the Hsp70 mRNA expression were 0.24±0.11 and 0.42±0.13 respectively in COPD patients and healthy smoking controls with the difference being significant (P<0.01). There was also significant difference in the A values of the Hsp70 expression between COPD patients and healthy smoking controls (20.9±9.9 vs 44.8±15.3, P<0.01). The levels of Hsp70 mRNA had strongly positive correlation with Hsp70 protein (r = 0.85, P<0 01). It was suggested that the expression of Hsp70 mRNA was in concordance with the expression of Hsp70, which could provide a basis on the study of Hsp70 gene regulation and Hsp70 gene in the development of COPD.

Expression of Peroxiredoxins and Pulmonary Surfactant Protein A Induced by Silica in Rat Lung Tissue

Silicosis is one of the most serious occupational diseases in China and dates back to centuries ago. In this study, we successfully established a rat model of silicosis by intratracheal silica injection for 28 days and determined hydroxyproline levels to evaluate collagen metabolism in lung homogenates. Oxidative stress status was evaluated by detecting catalase and glutathione peroxidase activities.

Forkhead box protein A2 and T helper type 2-mediated pulmonary inflammation

The transcription factor forkhead box protein A2(FOXA2, also known as hepatocyte nuclear factor 3β or transcription factor 3β), has been found to play pivotal roles in multiple phases of mammalian life, from the early development to the organofaction, and subsequently in homeostasis and metabolism in the adult. In the embryonic development period, FOXA2 is require d for the formation of the primitive node and notochord, and its absence results in embryonic lethality. Moreover, FOXA2 plays an important role not only in lung development, but also in T helper type 2(Th2)-mediated pulmonary inflammation and goblet cell hyperplasia. In this article, the role of FOXA2 in lung development and Th2-mediated pulmonary inflammation, as well as in goblet cell hyperplasia, is reviewed. FOXA2 deletion in airway epithelium results into Th2-mediated pulmonary inflammation and goblet cell hyperplasia in developing lung. Leukotriene pathway and signal transducers and activators of transcription 6 pathway may mediate this inflammation through recruitment and activation of denditric cell during lung developments. FOXA2 is a potential treatment target for lung diseases with Th2 inflammation and goblet cell hyperplasia, such as asthma and chronic obstructive pulmonary disease.

C-Reactive Protein as a Triage Test in Guiding Who Should Get a Confirmatory Test for Pulmonary Tuberculosis Diagnosis among Adults: A Case-Control Proof-of-Concept Study from Urban Tanzania

Background: The current screening tools for tuberculosis (TB) are inadequate resulting in insufficient TB case detection and continued community transmission of TB. As the world is geared into finding missing TB cases, new strategies are called for to aid in rapid identification of TB cases. This study aimed to evaluate the role C-reactive protein (CRP) in triaging patients to get a definitive test for active pulmonary TB diagnosis in urban Tanzania. Methods: A case-control study was conducted among pulmonary TB (PTB) patients and contacts without active PTB. The diagnosis of PTB was performed using GeneXpert MTB/RIF assay and culture. Blood was collected from cases and controls for measuring CRP levels during recruitment. We compared socio-demographic characteristics, clinical and laboratory parameters obtained during recruitment and performed diagnostic accuracy analyses for CRP. Results: Out of all 193 study participants who were involved in final analysis, 147 (76.2%) were males. Pulmonary TB cases had significantly lower median BMI than controls (median 17.4 kg/m2 [IQR: 15.8 - 19.2 kg/m2] vs., 24.9 kg/m2 [IQR: 22.1 - 28.5 kg/m2), p < 0.001). There was no statistical difference in prevalence of HIV between PTB cases and controls i.e., 13.33% vs., 11.7%, p = 0.48. CRP was significantly higher in PTB cases vs., controls (median 67.8 mg/L, [IQR: 36.5 - 116.9 mg/L] vs., 1.55 mg/L, [IQR: 0.59 - 6.0 mg/L], p = 0.003). Furthermore, CRP at cut-off ≥10 mg/L was associated with best combination of sensitivity, specificity and area under the curve of 89.9%, 95% CI: 82.2 - 95.0, 80.9%, CI: 71.4 - 88.2 and 0.85, 95% CI: 0.80 - 0.90 respectively. A multivariate logistic regression model adjusted for fever, night sweats and body mass index showed that CRP above 10 mg/L was significantly associated with PTB, aOR 5.2, 95% CI 1.2 - 22.8. Conclusions: CRP at cut-off ≥10 mg/L can be used to screen pulmonary TB. These findings can be used to improve TB screening algorithm by incorporating CRP in combination with TB symptoms to identify patients who need further confirmatory TB tests. However, additional prospective studies are required to support our findings and contribute into policy recommendations on use of CRP in a screening algorithm for pulmonary TB.

Effects of Huang Qi Wu Wu Decoction on Plasma Proteins in 70 Cases of Chronic Pulmonary Heart Disease

Simple immune agar diffusion test was used to assay the contents of 12 plasma proteins in 70 cases of chronic pulmonary heart disease treated by Huang Qi Wu Wu Decoction (黄芪五物汤), with the other 70 cases who were not given Huang Qi Wu Wu Decoction as the control group. The total clinical effective rate in the treatment group was 90.0%, while that in the control group was 75.7%, with a statistically significant difference between the two groups (P<0.05). In the treatment group, the levels of prealbumin, transferrin and fibronectin elevated obviously after treatment, and the contents of C-reactive protein, ceruloplasmin, haptoglobin, a 1-antitrypsin and a 1-acid glycoprotein decreased markedly (P<0.01). In the control group, only the levels of ceruloplasmin and C-reactive protein decreased significantly (P<0.05). It is shown that Huang Qi Wu Wu Decoction may enhance the therapeutic effects for pulmonary heart disease, regulate the metabolism of plasma proteins, and improve the life quality of the patients.

Diagnostic accuracy of C-reactive protein for bacterial infections in acute exacerbation of chronic obstructive pulmonary disease: A meta-analysis

Objective:The clinical diagnostic accuracy of C-reactive protein(CRP)for bacterial infections in acute exacerbation of chronic obstructive pulmonary disease(AECOPD)is still controversial.This study evaluated its accuracy through Meta-analysis.Methods:Studies on the diagnostic value of CRP for bacterial infections in AECOPD were searched form Web of Science,PubMed,Cochrane library,EMBASE,WANFANG DATA,CNKI and China Biology Medicine disc.The included studies were evaluated according to QUADAS-2 assessment tool.Stata 12.0 software was used for meta analyses to summarize the sensitivity and the specificity of the included studies,and the heterogeneity test was conducted.The symmetric receiver operating characteristic curves(SROC)was drawn and the area under the curve(AUC)was calculated,and the funnel plot was drawn to determine publication bias.Results:664 studies were initially identified,20 of which met the inclusion criteria(9 in English and 11 in Chinese).Meta-analysis results showed that the summary sensitivity and specificity of CRP in diagnosing bacterial infections in AECOPD were 0.84,95%CI(0.77-0.90)and 0.76,95%CI(0.67-0.82),respectively.The AUC of SROC was 0.87,95%CI(0.83-0.89).Conclusion:CRP has high accuracy,sensitivity and specificity in distinguishing AECOPD bacterial infections from non-bacterial infection.

Correlation of neonatal pulmonary surfactant protein A gene polymorphism with pneumonia susceptibility and inflammatory response

Objective:To study the correlation of neonatal pulmonary surfactant protein A gene polymorphism with pneumonia susceptibility and inflammatory response.Methods:Neonates who were born and diagnosed with pneumonia in Zigong Maternity and Child Healthcare Hospital between September 2015 and February 2017 were selected as pneumonia group, and neonates without infection were selected as control group. SP-A gene rs1059054 and rs1136454 loci polymorphism, the contents of inflammatory cytokines in serum as well as the expression of inflammatory transcription factors in peripheral blood were determined.Results:The constituent ratio of rs1059054 loci CC genotype of pneumonia group was significantly higher than that of control group while the constituent ratio of CT and TT genotypes were significantly lower than those of control group;the constituent ratio of rs1136454 loci AA genotype was significantly lower than that of control group while the constituent ratio of AG and GG genotypes were significantly higher than those of control group. PCT, sTREM1, TNF-α and IL-6 levels in serum as well as RORγt mRNA expression in peripheral blood of pneumonia children with SP-A gene rs1059054 loci CC genotype were significantly higher than those of pneumonia children with CT genotype and TT genotype while SOCS1 and Foxp3 mRNA expression in peripheral blood were significantly lower than those of pneumonia children with CT genotype and TT genotype;PCT, sTREM1, TNF-α and IL-6 levels in serum as well as RORγt mRNA expression in peripheral blood of pneumonia children with SP-A gene rs1136454 loci AA genotype were significantly lower than those of pneumonia children with AG genotype and GG genotype while SOCS1 and Foxp3 mRNA expression in peripheral blood were significantly higher than those of pneumonia children with AG genotype and GG genotype.Conclusion: Neonatal SP-A gene rs1059054 loci CC genotype can increase the pneumonia susceptibility and aggravate inflammatory response, and rs1136454 loci AA genotype can decrease pneumonia susceptibility and relieve inflammatory response.

Changes of Tumor Necrosis Factor, Surfactant Protein A, and Phospholipids in Bronchoalveolar Lavage Fluid in the Development and Progression of Coal Workers’ Pneumoconiosis

Objective To evaluate the alterations of biomarkers in the development and progression of coal workers＇ pneumoconiosis （CWP）. Methods The type and number of cells, and the levels of tumor necrosis factor-alpha （TNF-α）, pulmonary surfactant protein, phospholipids and fibronectin in bronchoalveolar lavage fluid were assayed in 14 health active coal miners, 21 coal miners without CWP and 13 miners with CWP of 0/1 to 1/1. Results Compared to active coal miners without CWP （8,23μg/mL）, TNF-α concentration was gradually decreased when dust exposure was stopped （5.90 μg/mL）. Elevated surfactant protein A （SP-A） level and phosphatidylglycerol （PG） to phosphatidylinositol （PI） ratio were found in miners actively exposed to coal dust （6528 μg/mL for SP-A and 10. for PG/P1）, and both parameters decreased when CWP progressed from CWP （0/1） （3419μg/mL for SP-A and 5.9 for PG/PI） to CWP （1/1） （1654 μg/mL for SP-A and 5.5 for PG/PI）. Conclusion Biomarkers in bronchoalveolar lavage fluid can be used to screen coal miners at high risk of developing coal workers＇ pneumoconiosis.

肌微管素相关微蛋白7在小鼠肺动脉高压中的作用及机制

目的探讨肌微管素相关微蛋白7(myotubularin related protein 7,MTMR7)在肺动脉高压中的作用及机制。方法健康♂小鼠、Mtmr7转基因(Mtmr7-Tg)小鼠各20只,分为对照组、Mtmr7-Tg组、野百合碱(monocrotaline,MCT)组、MCT+Mtmr7-Tg组,超声测量肺动脉血流加速时间(pulmonary artery acceleration time,PAT)和射血时间(pulmonary artery ejection time,PET),取材时分离右心室游离壁,计算右心肥厚指数,免疫染色观察肺小动脉重构。将小鼠肺动脉平滑肌细胞(pulmonary arterial smooth muscle cells,PASMCs)以低氧处理,观察其增殖和迁移情况。结果MTMR7在肺血管中表达,并且MCT处理之后,Mtmr7-Tg小鼠的PAT/PET比值降低(P<0.05),右心肥厚指数升高(P<0.01)。以Mtmr7基因过表达腺病毒转染PASMCs,可以抑制其增殖和迁移。用chemerin-9恢复p-ERK1/2的活性,过表达MTMR7对PASMCs增殖、迁移抑制作用消失。结论过表达MTMR7可减轻低氧诱导的小鼠PASMCs增殖和迁移,从而逆转肺动脉高压,机制与MTMR7降低ERK1/2磷酸化密切相关。

基于HMGB1介导的肺动脉平滑肌细胞焦亡探讨复方葶苈子汤治疗COPD-PH的作用机制

目的研究高迁移率族蛋白B1(high mobility group box 1,HMGB1)介导的肺动脉平滑肌细胞焦亡对慢性阻塞性肺疾病相关性肺动脉高压(chronic obstructive pulmonary disease-associated pulmonary hypertension,COPD-PH)形成的影响,及复方葶苈子汤的干预机制。方法以CCK-8法检测大鼠肺动脉平滑肌细胞(pulmonary arterial smooth muscle cells,PASMCs)的活性,筛选出合适的香烟烟雾提取物(cigarette smoke extract,CSE)和脂多糖(lipopolysaccharide,LPS)干预浓度及时间用于建立细胞损伤模型。将对数生长期细胞分为对照组、CSE组、LPS组和CSE+LPS组,对照组加入细胞培养液培养细胞。CSE组加入含10%CSE的细胞培养液,LPS组加入含1μg/mL LPS的细胞培养液,CSE+LPS组加入含10%CSE和1μg/mL LPS的细胞培养液,培养细胞24 h后,采用CCK-8法检测细胞活性,实时荧光定量聚合酶链反应(real-time fluorescent quantitative PCR,qRT-PCR)检测细胞中mRNA表达,蛋白质印迹法(Western blot,WB)检测细胞中蛋白表达,酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)检测细胞上清液中炎症因子含量,流式细胞术检测细胞焦亡,透射电子显微镜观察细胞的超微结构。以CCK-8法检测PASMCs的活性,筛选出复方葶苈子汤含药血清和甘草酸的最佳干预浓度及时间。将对数生长期细胞分为正常组、模型组、含药血清组和抑制剂组,对照组加入细胞培养液培养细胞,模型组、含药血清组和抑制剂组加入含10%CSE和1μg/mL LPS的细胞培养液,培养细胞24 h后弃去原培养液。正常组和模型组孔中加入含10%空白血清的细胞培养液,含药血清组孔中加入含10%复方葶苈子汤含药血清的细胞培养液,抑制剂组孔中加入含150μg/mL甘草酸和10%空白血清的细胞培养液,再培养细胞24 h后,采用CCK-8法检测细胞活性,RT-PCR检测细胞中mRNA表达,WB检测细胞中蛋白表达,ELISA检测细胞上清液中炎症因子含量,流式细胞术检测细胞焦亡,透射电子显微镜观察细胞的超微结构。结果筛选出10%CSE和1μg/mL LPS用于建立细胞损伤模型,干预时间为24 h。CSE、LPS和CSE联合LPS可以使PASMCs活性下降;可以促进PASMCs中HMGB1、RAGE、Caspase-8和GSDMD mRNA表达,HMGB1、RAGE、pro Caspase-8、cleaved Caspase-8和GSDMD蛋白表达,促进PASMCs上清液中白细胞介素-1β(interleukin-1β,IL-1β)和白细胞介素-18(interleukin-18,IL-18)分泌,促进PASMCs焦亡发生。筛选出10%复方葶苈子汤含药血清和150μg/mL甘草酸用于细胞干预,干预时间为24 h。复方葶苈子汤含药血清可以使损伤PASMCs活性升高;可以抑制损伤PASMCs中HMGB1、RAGE、Caspase-8和GSDMD mRNA表达,HMGB1、RAGE、pro Caspase-8、cleaved Caspase-8和GSDMD蛋白表达,抑制损伤PASMCs上清液中IL-1β和IL-18分泌,抑制PASMCs焦亡发生。结论复方葶苈子汤可以通过抑制HMGB1表达,抑制Caspase-8介导的PASMCs焦亡,并抑制炎症级联反应,从而改善COPD-PH。

氧浓度变化对人肺动脉内皮细胞中ANGPTL8表达的影响

目的探讨氧浓度变化对肺动脉内皮细胞(HPAECs)中血管生成素样蛋白8(ANGPTL8)表达的影响,以及ANGPTL8在肺动脉高压(PH)中的作用和机制。方法低氧(1%)和高氧(85%)处理HPAECs,Western blot和PCR等检测ANGPTL8的表达,并分析内皮-间质转化(EndMT)及ERK信号通路的变化。同时,高氧处理新生大鼠,构建支气管肺发育不良(BPD)模型,观察肺组织中ANGPTL8蛋白表达及ERK信号通路的变化。结果低氧组HPAECs中ANGPTL8的蛋白表达显著升高,伴随ERK信号通路的抑制(P<0.05)。ANGPTL8促进低氧诱导的EndMT过程(P<0.05),沉默ANGPTL8可以部分逆转EndMT过程。高氧组HPAECs和大鼠肺组织中ANGPTL8蛋白表达降低,伴随ERK信号通路的激活(P<0.05)。结论HPAECs中ANGPTL8对氧浓度变化高度敏感,其表达的变化与ERK信号通路的活性状态密切相关,对PH的发展具有潜在的调控作用。

血清FSTL1、GRP78对老年COPD患者疾病严重程度及再入院的评估价值

目的探讨血清卵泡抑素样蛋白1(FSTL1)、葡萄糖调节蛋白78(GRP78)对老年慢性阻塞性肺疾病(COPD)患者疾病严重程度及再入院的评估价值。方法选取该院于2020年3月至2023年5月收治的100例老年COPD患者作为COPD组,根据病情严重程度分为Ⅰ级(35例)、Ⅱ级(46例)、Ⅲ级(19例)。另选取同期体检的100例老年志愿者作为对照组。采用酶联免疫吸附试验(ELISA)检测两组血清FSTL1、GRP78水平。采用Spearman和Pearson相关性分析血清FSTL1、GRP78水平与一般资料的相关性,采用受试者工作特征(ROC)曲线分析COPD患者血清FSTL1、GRP78水平对疾病严重程度及再入院的评估价值。结果与对照组相比,COPD组血清FSTL1、GRP78水平均显著升高(P<0.05)。COPD患者血清FSTL1、GRP78水平随着病情严重程度增加而依次升高(P<0.05)。与非再入院组相比,再入院组患者血清FSTL1、GRP78水平显著升高(P<0.05)。Spearman相关性分析显示,血清FSTL1、GRP78水平与吸烟史、高血压史、病情严重程度呈正相关(P<0.05);Pearson相关性分析显示,血清FSTL1、GRP78水平与动脉血氧分压(PaO 2)、第1秒用力呼吸容积与用力肺活量的比值(FEV 1/FVC)、第1秒用力呼气容积占预计值百分比(FEV 1%pred)呈负相关(P<0.05),与二氧化碳分压(PaCO 2)呈正相关(P<0.05)。ROC曲线分析显示,COPD患者血清FSTL1、GRP78及二者联合评估病情严重程度及再入院的曲线下面积(AUC)均较高,并且二者联合明显均优于单独评估(P<0.05)。结论COPD患者血清FSTL1、GRP78水平均明显升高,二者与病情严重程度相关,对病情严重程度及再入院的评估价值较高。

钙敏感受体在肺动脉高压小鼠肺静脉重塑中的作用及机制研究

目的研究钙敏感受体(CaSR)在肺动脉高压(PH)小鼠肺静脉重塑中的作用及机制。方法选用C57/BL6小鼠28只,随机均分为常氧(Nor)组、常氧+NPS2143(Nor+NPS)组、低氧联合SU5416(HySu)组、低氧联合SU5416+NPS2143(HySu+NPS)组,每组7只。通过低氧(10%O 2)条件下饲养加皮下注射SU5416的方法构建PH小鼠模型。利用血流动力学测定和右心室肥厚指数(RVHI)分析评估PH,应用Weigert弹力纤维染色、HE染色、α-SMA免疫组化染色和CaSR免疫组化染色评估血管重塑。通过Western blot检测肺静脉CaSR、经典瞬时受体电位蛋白(TRPC)6及TRPC4的表达水平。结果通过低氧联合SU5416成功构建PH小鼠模型。HySu组小鼠右心室收缩压(RVSP)和RVHI显著高于Nor组。PH小鼠的远端肺静脉发生血管重塑,但近端肺静脉无血管重塑。PH小鼠的CaSR、TRPC6表达水平较Nor组显著上调(P<0.05),而TRPC4表达与Nor组比较无显著差异(P>0.05)。加入NPS2143干预后,HySu+NPS组小鼠的RVSP和RVHI显著低于HySu组(P<0.05),并且NPS2143可显著缓解低氧联合SU5416诱导的小鼠远端肺静脉重塑,并下调PH小鼠远端肺静脉CaSR和TRPC6表达(P<0.05)。结论CaSR在PH小鼠远端肺静脉重塑中发挥核心作用,低氧联合SU5416可能通过上调远端肺静脉平滑肌CaSR表达而引起TRPC6表达上调导致肺静脉重塑。

重组结核分枝杆菌融合蛋白皮肤试验在肺结核诊断中诊断价值的研究

目的:针对肺结核的诊断,引入重组结核分枝杆菌融合蛋白皮肤试验,评估其临床价值。方法:回顾性分析2022年3月—2024年3月麻城市人民医院收治的236例疑似肺结核患者的临床资料作为研究样本,所有患者均进行重组结核分枝杆菌融合蛋白皮肤试验,参照结核分枝杆菌痰涂片或者痰培养结果(金标准)对重组结核分枝杆菌融合蛋白皮肤试验的诊断结果进行分析,并计算其诊断效能,同时进行成本效益分析。结果:参考结核分枝杆菌痰培养结果,236例疑似结核病患者共确诊184例,(77.96%);非肺结核患者52例(22.03%);重组结核分枝杆菌融合蛋白皮肤试验结果中,共检出结核病患者159例,占比67.37%,非结核病患者77例,(32.62%);重组结核分枝杆菌融合蛋白皮肤试验诊断灵敏度85.86%(158/184)、特异度98.07%(51/52)、准确率88.55%(209/236),参照金标准,差异有统计学意义(P<0.05);重组结核分枝杆菌融合蛋白皮肤试验成本费用相对低廉,差异有统计学意义(P<0.05)。结论:在肺结核的临床诊断中,重组结核分枝杆菌融合蛋白皮肤试验具有诊断准确率高,操作简单,观察方便,成本价格低廉的优势,具有参考与应用价值。

PCT、WBC联合CRP在ICU呼吸感染中的表达、诊断价值及与CPIS评分的相关性分析

目的分析降钙素原(PCT)、白细胞计数(WBC)联合C反应蛋白(CRP)在重症监护室(ICU)呼吸感染中的表达、诊断价值及与临床肺部感染评分(CPIS)的相关性。方法选取2019年3月至2024年6月涿州市医院ICU呼吸感染患者105例为观察组,另外选取于ICU未发生呼吸感染117例患者为对照组。通过Zybio-Q7型免疫定量分析仪对两组PCT水平进行检测,通过XT-4000i型白细胞分析仪对两组WBC水平进行检测,通过免疫比浊法对两组CRP水平进行检测。分析PCT、WBC、CRP的表达变化与ICU呼吸感染临床特征、CPIS评分的关系及对ICU呼吸感染的诊断价值。结果观察组入住ICU时间(>15 d)和机械通气时间(>7 d)的占比显著高于对照组(P<0.05)。与对照组比较,观察组PCT、WBC、CRP水平和CPIS评分升高,差异具有统计学意义(P<0.05)。多因素Logistic回归分析结果显示,入住ICU时间>15 d(OR=4.087)、PCT(OR=6.543)、WBC(OR=2.652)和CRP(OR=8.964)为影响ICU呼吸感染发生的危险因素。PCT、WBC、CRP与CPIS评分之间Pearson相关性分析显示,PCT、CPIS评分之间正相关(r=0.925,P=0.001);WBC、CPIS评分之间正相关(r=0.739,P=0.001);CRP、CPIS评分之间正相关(r=0.948,P=0.001)。ROC曲线显示,三项联合对ICU呼吸感染的诊断价值高于PCT、WBC、CRP单项诊断(P=0.002)。结论PCT、WBC联合CRP对ICU呼吸感染的诊断价值较高,且与CPIS评分呈正相关,因此上述指标可为ICU呼吸感染的早期诊断提供可靠依据。

hs-CRP与白蛋白比值对呼吸衰竭COPD患者撤机失败的预测价值

目的 通过探讨超敏C反应蛋白(hs-CRP)与白蛋白(Alb)与呼吸衰竭慢性阻塞性肺疾病(COPD)患者撤机失败的关系,分析其对该类患者撤机失败的预测价值。方法 选取2022年10月—2023年10月浙江省人民医院淳安分院收治的242例呼吸衰竭COPD患者,根据撤机结果将患者分为2组:撤机成功组(180例)与撤机失败组(62例)。采用单因素分析比较2组患者的一般资料,采用多因素logistic回归分析法分析呼吸衰竭COPD患者撤机失败的影响因素,绘制ROC曲线分析hs-CRP与Alb比值对呼吸衰竭COPD患者撤机失败的预测价值。结果 撤机成功组患者年龄、急性生理与慢性健康评分(APACHEⅡ)、hs-CRP、hs-CRP与Alb比值、胃肠功能受损发生率较撤机失败组低,机械通气时间较撤机失败组短,Alb水平较撤机失败组高(P<0.05);logistic回归分析结果显示,年龄较大、APACHEⅡ评分高、hs-CRP与Alb比值较高均为呼吸衰竭COPD患者撤机失败的危险因素;ROC曲线结果显示,年龄、APACHEⅡ、hs-CRP与Alb比值预测呼吸衰竭COPD患者撤机失败的AUC为0.735、0.716、0.927,其中hs-CRP与Alb比值的预测价值最高。结论 呼吸衰竭COPD患者撤机失败的发生与hs-CRP与Alb比值的上升有关,其可以作为撤机失败发生的预测因子。