ObjectiveTo explore the expression and clinical significance of Ras-related C3 botulinum toxin substrate 1 (Rac-1) protein and hypoxia-inducible factor 1α (HIF-1α) in breast cancer and their relationship with tumor stage, node metastasis status and hormone receptor status. MethodsImmunohistochemical method was used to detect the expression of Rac-1 and HIF-1α protein in 139 specimens of breast cancer,combined with their clinical pathological characteristics and hormone receptor status for statistical analysis.Results The expression of Rac-1 and HIF-1α in breast cancer tissue were 64.75 ％ and 65.47 ％,respectively.The levels were related with the stage of TNM and histological grade (P ＜ 0.05,P ＜ 0.01).There was no significant correlation between these two proteins'expression and the age of patient,primary pathological location, histologic type and menstruation status of breast cancer patients(P ＞ 0.05). The expression of Rac-1 and HIF-1α had no statistical significance between triple negative breast cancer(TNBC)and non-triple negative breast cancer (NTNBC) patients (x2 =1.1229,x2 =0.1786,P ＞ 0.05); The expression level of Rac-1 was positively correlated with HIF-1α in these breast cancer specimens (rs =0.414,P ＜ 0.01).ConclusionThe expression of Rac-1 is positively correlated with HIF-1α level in breast cancer.The relationship between Rac-1 and HIF-1α might be as a new important marker to estimate the invasion,metastasis extent and prognosis in breast cancer.Blocking the regulation between Rac-1 and HIF-1α might be a new treatment target in breast cancer.

Objective To construct and express human papillomavirus type 11(HPV11) E7 gene with recombinant adenovirus vectors. Methods HPV11 E7 gene was amplified by PCR and directionally cloned into vector pENTR-TOPO to form TOPO-E7 plasmid. E7 gene was transferred into the pAD/CMV/V5-DESTTM gateway vector by LR recombination reaction with pAD/CMV/V5-DESTTM gateway vectors and TOPO-E7 plasmid. The recombination vector was digested by Pac I enzyme and transfected into 293A cell by Lipofectamine method to obtain recombinant adenovirus vectors pAD-E7. Expression of E7 on HaCaT cells infected with pAD-E7 vectors was analyzed by confocal microscopy. Results The recombinant plasmid TOPO-E7 was identified and confirmed with enzyme digestion and sequencing. Recombinant adenovirus vectors pAD-E7 were generated efficiently with a titer of 1.4 ? 107 pfu/mL in transfected 293A cells. E7 protein could be identified in HaCaT cells with confocal microscope 48 h after infected with recombinant adenovirus vector. Conclusions The results indicate efficient expression of HPV11 E7 gene in eukaryotic cells by recombinant adenovirus mediated transfer, which facilitates further research of its function.

Objective To explore the manifestations and diagnostic value of CT and MRI in gynecological acute abdomen.Methods CT and MRI features of 45 patients with gynecological acute abdomen proved by surgery were reviewed,including 17 ectopic pregnancy,10 ovarian cyst rupture,6 pedicle torsion of ovarian tumors,9 tubo-ovarian abscess and pelvic inflammation and 3 uterine perforation.31 cases underwent CT examination,14 cases underwent MRI examination and 39 cases underwent non-enhancement and enhancement scanning.Results In the 17 ectopic pregnancy cases,cystic structure surrounded by a thick wall or heterogeneous mass located in adnexal area was found,with 10 cases showing increased number of peripheral vessels penetrating into tumors.In the 10 cases with rupture of ovarian cyst, thick capsule wall was incomplete and cystic cavities collapsed in 7 lesions.In the 6 cases with pedicle torsion of ovarian tumors,irregular thickening of tumor pedicle were found in 4 cases,ovarian enlargement with surrounding follicles arrayed as fruit platter were found in 2 cases.In the 9 cases with tubo-ovarian abscess and pelvic inflammation,honeycomb-shaped lesions and multilocular changes were found in 6 cases and fallopian tube expanded like sausages in 5 cases.3 cases with uterine perforation exhibited intrauterine gas and fluid.Uterus with overflowed gas was found in 2 cases.One case had the thin and incomplete uterine wall.Conclusion CT and MRI can be used to confirm the causes of gynecological acute abdomen,and to comprehensively display the anatomical structures and pathological changes of pelvic tissues and organs.Therefore,CT and MRI are effective supplement means for the clinical diagnosis of gynecological acute abdomen.

Objective To investigate the expression of miR-124 in glioblastoma (GBM) cell lines LN229 and LN229R,as well as the regulatory mechanism of miR-124 on radiosensitivity of LN229R cells.Methods miR-124 mimic (miR-124) and negative control (miR-NC),STAT3 overexpression plasmid (STAT3) and pcDNA3.1 vector (pcDNA) were transfected or co-transfected into radioresistant glioma cells LN229R.qRT-PCR was employed to analyze the expression of miR-124 in LN229 and LN229R cells.The survival rate and sensitivity-related parameters of LN229R cells at different doses were analyzed by cloning formation assay.Cell apoptosis of LN229R was evaluated by flow cytometry.Targeting gene of miR-124 was predicted using Targetscan software and verified by the double-luciferase reporter assay.Western blot assay was performed to detect STAT3 protein expression.Results The expression of miR-124 in LN229R cells (0.32 ± 0.03) was significantly lower than that in LN229 cells (1.02 ± 0.09) (t =12.780,P＜0.05).Transfection of miR-124 mimics promoted the expression of miR-124 in LN229R cells (4.02±0.39) compared with miR-NC group (0.95±0.06) (t=13.476,P＜0.05).After 8 Gy irradiation,the survival rate of LN229R cells transfected with miR-124 mimics (0.003 ± 0.000 4) was significantly lower than that in miR-NC group (0.033±0.005 0) (t=5.655,P＜0.05),and the apoptosis rate (22.34±2.42) ％ was significantly higher than that in miR-NC group (4.69 ± 0.51) ％ (t =12.361,P＜0.05).STAT3 was identified to be a target gene of miR-124.Exogenous restoration of STAT3 reversed the inhibitory effect of miR-124 on LN229R cell survival.Conclusion miR-124 increases the radiosensitivity of LN229R cells by targeting STAT3.

Acute lung injury/acute respiratory distress syndrome (ALI/ARDS) is characterized by diffuse alveolar injury primarily caused by an excessive inflammatory response. Regrettably, the lack of effective pharmacotherapy currently available contributes to the high mortality rate in patients with this condition. Xuebijing (XBJ), a traditional Chinese medicine recognized for its potent anti-inflammatory properties, exhibits promise as a potential therapeutic agent for ALI/ARDS. This study aimed to explore the preventive effects of XBJ on ALI and its underlying mechanism. To this end, we established an LPS-induced ALI model and treated ALI mice with XBJ. Our results demonstrated that pre-treatment with XBJ significantly alleviated lung inflammation and increased the survival rate of ALI mice by 37.5%. Moreover, XBJ substantially suppressed the production of TNF-α, IL-6, and IL-1β in the lung tissue. Subsequently, we performed a network pharmacology analysis and identified identified 109 potential target genes of XBJ that were mainly involved in multiple signaling pathways related to programmed cell death and anti-inflammatory responses. Furthermore, we found that XBJ exerted its inhibitory effect on gasdermin-E-mediated pyroptosis of lung cells by suppressing TNF-α production. Therefore, this study not only establishes the preventive efficacy of XBJ in ALI but also reveals its role in protecting alveolar epithelial cells against gasdermin-E-mediated pyroptosis by reducing TNF-α release.
Animals ; Mice ; Alveolar Epithelial Cells ; Pyroptosis ; Gasdermins ; Lipopolysaccharides/adverse effects* ; Tumor Necrosis Factor-alpha ; Acute Lung Injury/drug therapy* ; Respiratory Distress Syndrome