目的：探讨Na+，K+-ATP1b1（ATP1b1）对肝癌细胞生长及侵袭能力的影响。方法：将ATP1b1表达质粒ATP1b1-CMV-FLAG转染肝癌细胞SMMC-7721，以空质粒转染作对照，通过RT-PCR测定ATP1b1的mRNA表达，在光镜下观察细胞形态学变化，MTT法检测细胞生长增殖，Transwell细胞侵袭实验分析肝癌细胞的侵袭能力。结果：ATP1b1-CMV-FLAG转染后，SMMC-7721的ATP1b1 mRNA表达水平明显增高（P＜0.05）,转染ATP1b1-CMV-FLAG组、pFLAG-CMV-1组和脂质体组的ATP1b1 mRNA表达水平分别为1.159、0.182和0.093;转染ATP1b1-CMV-FLAG的SMMC-7721细胞呈变性改变，其生长受到抑制，转染相同DNA浓度的ATP1b1-CMV-FLAG组与pFLAG-CMV-1组对细胞生长的抑制效应有显著性差异（P＜0.05）；Transwell细胞侵袭实验显示转染ATP1b1-CMV-FLAG组的SMMC-7721细胞的体外侵袭力明显受抑制（P＜0.01）。结论：实验结果说明了促进ATP1b1表达能抑制肝癌细胞的生长及侵袭行为。
自1987年Mouret完成世界首例腹腔镜胆囊切除术以来，随着外科医生手术技术的不断提高和腹腔镜器械的逐渐改进，腹腔镜微创技术的应用范围越来越广泛，以腹腔镜为代表的微创外科已经成为21世纪外科发展的方向之一，其对胃肠道恶性肿瘤根治的可行性和手术安全性已经得到认可。已有多项RCT研究显示，腹腔镜结直肠癌手术与开腹手术具有相当的近、远期疗效，美国结直肠癌外科医师协会已将其列为治疗结直肠癌的标准手术方式之一。近年来，腹腔镜在胃癌根治术中的应用已逐渐由早期胃癌扩展到进展期胃癌，并取得了与开腹手术相当的近、中期疗效。但是人们对CO2 气腹是否有利于胃肠道肿瘤的侵袭、转移一直心存疑虑，随之一些有关腹腔镜技术中不同种类、不同压力的气腹与胃肠道肿瘤侵袭、转移关系的研究报道相继出现,不同学者报道结果有较大差异，有些学者认为CO2气腹有利于胃肠道肿瘤的侵袭、转移; 而有些学者却认为CO2气腹对胃肠道肿瘤的侵袭、转移无显著影响。..................
ObjectiveTo investigate the regulatory mechanism of thioredoxin binding protein (TXNIP)/nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) pathway in the occurrence and development of breast cancer.MethodsThe resected 15 cases of breast cancer tissues and their adjacent tissues in our hospital from September 2019 to June 2020 were selected, and the immunohistochemistry was used to detect the expression levels of TXNIP and NLRP3 in breast cancer and its adjacent tissues. Three kinds of breast cancer cell lines (MDA-MB231, MCF-7 and SKBR3) and normal breast epithelial cell line (HMEC) were collected. Western blot was used to detect the relative expression levels of TXNIP and NLRP3 in three kinds of breast cancer cell lines and HMEC cell line. MDA-MB231 cancer cells were divided into blank control group (normal culture without any treatment), TXNIP overexpression group (Ad-TXNIP group, transfected with adenovirus vector carrying TXNIP overexpression sequence), Ad-TXNIP negative control group (Ad-eGFP1 group, transfected of empty adenovirus vector without TXNIP overexpression sequence), NLRP3 overexpression group (Ad-NLRP3 group, transfected with adenovirus vector containing NLRP3 overexpression sequence), TXNIP and NLRP3 overexpression co-transfection group (Ad-TXNIP+Ad-NLRP3 group, co-transfection of adenovirus vector carrying TXNIP and NLRP3 overexpression sequence), TXNIP overexpression and Ad-NLRP3 negative control (Ad-eGFP2) co-transfection group (Ad-TXNIP+Ad-eGFP2 group,co-transfection of adenovirus vector carrying TXNIP overexpression sequence and empty adenovirus without NLRP3 overexpression sequence). After 24 hours of transfection and culture, CCK-8 method was used to detect the MDA-MB231 cells proliferation. Transwell chamber method was used to detect MDA-MB231 cells migration and invasion. Nude mice tumorigenicity test was used to detect the tumorigenicity of the MDA-MB231 cells in vivo. Western blot was used to detect the expressions of TXNIP, NLRP3, proliferation marker protein (Ki-67), caspase-1, vascular endothelial growth factor (VEGF), interleukin (IL)-1β, IL-18 and caspase-1 precursor protein (pro-caspase-1) in the MDA-MB231 cells.ResultsCompared with the adjacent tissues, the relative expression level of TXNIP decreased (P<0.05) and the relative expression level of NLRP3 increased (P<0.05) in breast cancer tissues. Compared with normal breast epithelial cell line (HMEC cell line), the relative expression levels of TXNIP in MDA-MB231, MCF-7 and SKBR3 breast cancer cell lines were decreased (P<0.05), and the relative expression levels of NLRP3 were increased (P<0.05). Compared with the blank control group, the relative expression levels of TXNIP, NLRP3, IL-1β, IL-18, pro-caspase-1 and caspase-1 were increased (P<0.05), the relative expression levels of Ki-67 and VEGF, the proliferation activity, invasion and migration ability of MDA-MB231 cells and tumor weight were decreased (P<0.05) in the Ad-TXNIP group and the Ad-NLRP3 group. Compared with the Ad-TXNIP group and the Ad-NLRP3 group, the relative expression levels of TXNIP, NLRP3, IL-1β, IL-18, pro-caspase-1 and caspase-1 were further increased (P<0.05), the relative expression levels of Ki-67 and VEGF, the proliferation activity, invasion and migration ability of MDA-MB231 cells and tumor weight were further decreased (P<0.05) in the Ad-TXNIP+Ad-NLRP3 group.ConclusionsIn breast cancer tissues and breast cancer cell lines, TXNIP is low expression and NLRP3 is high expression. They can interact with each other to promote pyroptosis and inhibit the proliferation, invasion and migration of breast cancer cells.
目的 观察鼻内镜下三种不同微创手术治疗非侵袭型真菌性上颌窦炎的疗效及氟康唑冲洗术腔的临床意义。 方法 回顾性分析我科2006年1月－2010年12月收治的284例非侵袭型真菌性上颌窦炎住院患者资料。患者分别采用单纯鼻内镜下上颌窦窦口开放术（术式1）、鼻内镜下上颌窦窦口开放联合经唇龈沟上颌窦前壁开窗（术式2）、以及鼻内镜下上颌窦窦口开放联合下鼻道开窗（术式3）进行治疗；术式3治疗的患者术后定期换药时，分别使用生理盐水或氟康唑反复冲洗鼻腔和上颌窦。所有患者门诊随访至少半年。 结果 在本组接受术式1、术式2和术式3治疗的患者分别有51例、45例和188例。上述三种术式治疗的患者中，分别有15例，9例和6例患者出现复发，复发率分别为29.6%、20.0%和3.2%；其中术式3治疗的患者复发率显著性低于术式1或术式2治疗的患者（P＜0.05）。在术式3治疗的患者中，生理盐水和氟康唑冲洗的伤口愈合时间分别为3.8周和 3.7周，两种冲洗方式对伤口的愈合影响差异无统计学意义（P＞0.05）。 结论 鼻内镜下上颌窦窦口开放联合下鼻道开窗是治疗非侵袭型霉菌性上颌窦炎的最佳方式，且伤口愈合时间与冲洗液种类无关。
Objective To examine the effects of newly designed LY52 on the expression of matrix metalloproteinases and invasive ability of hepatocellular carcinoma HepG2 cells. Methods The effects of LY52 on the proliferations of HepG2 cells were detected by MTT assay. Gelatin zymography and Western blot were used to detect the effects of LY52 on matrix metalloproteinase-2 expression in the cell line. Transwell chamber assay was used to detect the effects of LY52 on the invasion of the cells. Results No obvious inhibitory or cytotoxicity effects of LY52 was found in lower concentrations (lt;200 μg/ml) of LY52. Gelatin zymography and Western blot showed that matrix metalloproteinase-2 expression were inhibited by LY52 in a dose-dependent manner in HepG2 cells. Furthermore, transwell chamber assay showed that LY52 could significantly inhibit the invasion of the cell line in a dose-dependent manner.Conclusion The results suggest that LY52 may inhibit the invasion of hepatocellular carcinoma cells by suppressing the matrix metalloproteinase-2 activity.
Objective To investigate the role of chemokine receptor CXCR7 in the development and progression of pancreatic carcinoma. Methods The short hairpin RNA (shRNA) targeting CXCR7 was designed and delivered into AsPC-1 pancreatic carcinoma cells to knock down CXCR7 expression. The cell proliferation, cell cycle, and apoptosis after CXCR7 knockdown was determined by MTT and flow cytometry, respectively. The invasive ability of pancreatic carcinoma cells was evaluated by using the Transwell system. Results Compared with the blank control group (BC group), transfection of AsPC-1 cells with CXCR7-shRNA resulted in a significantly decreased expression of CXCR7 at both mRNA and protein levels (P<0.05), and the ability of proliferation and invasion significantly decreased (P<0.05). Knockdown of CXCR7 also significantly increase apoptosis (P<0.05), induce cell cycle arrest at G0/G1 phase (P<0.05). Conclusions Taken together, the present study showes that the knockdown of CXCR7 expression may play an important role in pancreatic carcinoma development, invasion, and metastasis, CXCR7 may be a potential therapeutic target for the treatment of pancreatic carcinoma.
ObjectiveTo investigate the effects of pipecolic acid oxidase (PIPOX) on the proliferation, apoptosis, migration and invasion of primary liver cancer cells. MethodsImmunohistochemical staining and analysis of The Cancer Genome Atlas (TCGA) database were used to examine the PIPOX expression levels in liver cancer tissues and paired adjacent normal tissues, and studied their relationship with patient prognosis. Liver cancer cell lines stably overexpressing or knocking out PIPOX were constructed to explore PIPOX’s impact on liver cancer cell proliferation, apoptosis, migration and invasion by conducting in vitro functional experiments such as CCK-8, EdU, apoptosis detection, and Transwell assays. In vivo, nude mice subcutaneous tumor models and lung metastasis models were used to verify PIPOX’s effect on liver cancer growth and metastasis. Real-time quantitative polymerase chain reaction (RT-qPCR) and western blot were both employed to detect the expression of epithelial-mesenchymal transition (EMT) markers in liver cancer cells. ResultsImmunohistochemical staining and TCGA database analysis revealed that PIPOX expression was significantly lower in liver cancer tissues compared to paired adjacent normal tissues (P<0.05). Prognostic analysis indicated shorter overall survival and disease-free survival in PIPOX low expression group (P<0.05). In vitro gain- and loss-of-function experiments showed that PIPOX significantly inhibited liver cancer cell migration and invasion (P<0.05), while having no significant effects on their proliferation and apoptosis (P>0.05). Animal experiments also confirmed that PIPOX significantly inhibited liver cancer lung metastasis (P<0.05), but had no significant effects on tumor growth (P>0.05). Finally, RT-qPCR and western blot results revealed that PIPOX promoted the expression of the epithelial marker E-cadherin (P<0.05) and inhibited the expression of mesenchymal markers (N-cadherin, vimentin, Snail) (P<0.05). ConclusionsPIPOX significantly inhibits liver cancer cell migration and invasion, potentially via suppressing the EMT process. However, PIPOX does not significantly affect liver cancer cell proliferation and apoptosis.
Objective To summarize the clinical features, predisposing factors, diagnosis, therapeutic outcome, and prognosis of invasive pulmonary fungal infection( IPFI) . Methods 90 cases with pathologically proved IPFI, admitted in non-intensive care unit in Xiangya Hospital from January 2005 to February 2012, were retrospectively analyzed. Results The pathogenic examination revealed Aspergillosis in 56 cases( 62. 2% ) , Cryptococcus in 18 cases( 20. 0% ) , Mucormycosis in 6 cases( 6. 7% ) , and Histoplasma in 6 cases( 6. 7% ) , etc. The underlying diseases were reported in 87 cases, and mainly included COPD, pulmonary tuberculosis, and diabetes mellitus. Cough and expectoration were the common clinical symptoms. 49 patients ( 54. 4% ) received long-term and broad-spectrum antibiotic therapy. The CT results revealed masses type in 25 cases( 27. 8%) , nodule lesions type in 15 cases( 16. 7% ) , lung consolidation type in 22 cases( 24. 4% ) , cavity type in 22 cases( 24. 4% ) , aspergilloma type in 6 cases( 6. 7% ) . 47 patients were clinical diagnosed with IPFI before biopsy with preliminary diagnosis accordance rate of 52. 2% . 31 cases ( 34. 4% ) underwent surgical resection of pulmonary lesions, and no recurrence was detected over two-year follow up. 56 cases ( 62. 2% ) received systemic anti-fugal therapy, and 43 cases( 76. 8% ) were cured or significantly improved. 3 cases ( 3. 3% ) refused any therapy. Conclusions The most frequently isolated pathogen of IPFI is Aspergillosis. The mainly underlying diseases are COPD, pulmonary tuberculosis, and diabetes mellitus. Long-termand broad-spectrum antibiotic therapy may be the major risk factor. Pathological examination is needed for final diagnosis. Surgical procedure can achieve optimal prognosis.
Objective To explore the correlation between the quantitative and qualitative features of CT images and the invasiveness of pulmonary ground-glass nodules, providing reference value for preoperative planning of patients with ground-glass nodules. MethodsThe patients with ground-glass nodules who underwent surgical treatment and were diagnosed with pulmonary adenocarcinoma from September 2020 to July 2022 at the Third Affiliated Hospital of Kunming Medical University were collected. Based on the pathological diagnosis results, they were divided into two groups: a non-invasive adenocarcinoma group with in situ and minimally invasive adenocarcinoma, and an invasive adenocarcinoma group. Imaging features were collected, and a univariate logistic regression analysis was conducted on the clinical and imaging data of the patients. Variables with statistical difference were selected for multivariate logistic regression analysis to establish a predictive model of invasive adenocarcinoma based on independent risk factors. Finally, the sensitivity and specificity were calculated based on the Youden index. Results A total of 555 patients were collected. The were 310 patients in the non-invasive adenocarcinoma group, including 235 females and 75 males, with a meadian age of 49 (43, 58) years, and 245 patients in the invasive adenocarcinoma group, including 163 females and 82 males, with a meadian age of 53 (46, 61) years. The binary logistic regression analysis showed that the maximum diameter (OR=4.707, 95%CI 2.060 to 10.758), consolidation/tumor ratio (CTR, OR=1.027, 95%CI 1.011 to 1.043), maximum CT value (OR=1.025, 95%CI 1.004 to 1.047), mean CT value (OR=1.035, 95%CI 1.008 to 1.063), spiculation sign (OR=2.055, 95%CI 1.148 to 3.679), and vascular convergence sign (OR=2.508, 95%CI 1.345 to 4.676) were independent risk factors for the occurrence of invasive adenocarcinoma (P<0.05). Based on the independent predictive factors, a predictive model of invasive adenocarcinoma was constructed. The formula for the model prediction was: Logit(P)=–1.293+1.549×maximum diameter of lesion+0.026×CTR+0.025×maximum CT value+0.034×mean CT value+0.72×spiculation sign+0.919×vascular convergence sign. The area under the receiver operating characteristic curve of the model was 0.910 (95%CI 0.885 to 0.934), indicating that the model had good discrimination ability. The calibration curve showed that the predictive model had good calibration, and the decision analysis curve showed that the model had good clinical utility. Conclusion The predictive model combining quantitative and qualitative features of CT has a good predictive ability for the invasiveness of ground-glass nodules. Its predictive performance is higher than any single indicator.