Objective To investigate the effects of Gli2 on the proliferation,growth,migration,and invasion of oral cancer cells(Tca8113)at the cellular level,and to clarify the molecular mechanism of how Gli2 regulation affects the migration and invasion of oral cancer cells.Methods Small interfering(si)RNA was used to inhibit Gli2 expression in Tca8113 cells.The effects of Gli2 on the proliferation,growth,migration,and invasion of Tca8113 cells were examined by CCK-8,platb cloning,and transwell chamber assay.Further qRT-PCR and Western blot assays were used to explore the mechanism of how Gli2 regulation effects the malignant proliferation and metastasis of Tca8113 cells.Results The mRNA and protein expression of Gli2 in oral cancer cells(Tca8113)increased.Interference of Gli2 expression inhibited the proliferation,growth,migration,and invasion of Tca8113 cells.Further experiments showed that interfering with Gli2 expression inhibited the mRNA and protein expression of key factors in the Hedgehog(Hh)pathway.In addition,interference of Gli2 expression significantly affected the mRNA and protein expression of key factors in epithelial mesenchymal transformation(EMT)pathways.Conclusions Gli2 is abnormally activated during oral cancer,and interference of Gli2 expression significantly inhibits the proliferation,growth,migration,and invasion of oral cancer cells.Gli2 influences the migration and invasion of oral cancer cells by regulating the Hh and EMT pathways.This study has provided a new way to elucidate the pathogenesis of oral cancer and new perspectives on the clinical treatment of oral cancer.

Immunodeficient animal models play an important role in preclinical research and are important experimental tools in modern biomedical research that are widely used in immunology,genetics,oncology,microbiology,and other research fields.Gene editing is a technology for targeted modification of biological genomes.From emergence to application,it has greatly promoted the development of biomedical research.Gene editing technology mainly includes homing endonucleases,zinc finger nucleases,transcription activator-like effector nucleases,and the CRISPR/Cas9 system.Researchers have used these technologies to establish various types of immunodeficient animal models,each with advantages and limitations.In recent years,a large number of studies have confirmed that the human immunodeficient animal model accurately simulates the functions of cancer cells,drugs,and the human immune system,better simulates human diseases,and is widely used to study human immunobiology and the potential mechanisms of complex diseases.In this article,we review the progress in the research and application of gene editing technology to the establishment of immunodeficient animal models,discuss in depth the problems and optimization strategies of gene editing technology in the preparation of immunodeficient animal models,and present its future development prospects to provide references for researchers to select and establish immunodeficient animal models.

Objectives:To explore the changes of some peripheral blood cells related to inflammation in patients with non-arteritis central retinal artery occlusion (NA-CRAO).Methods:A retrospective clinical study. From July 2019 to July 2021, a total of 218 patients with NA-CRAO hospitalized (NA-CRAO group) in Department of Ophthalmology, Xi'an People's Hospital (Xi'an Fourth Hospital) and 218 patients with routine physical examination (control group) during the same period were included in the study. There were no significant differences in age ( t=0.60), sex composition ratio ( χ2=0.83) and body mass index ( t=0.77) between the two groups ( P>0.05). 0.2 ml fasting peripheral blood was collected from the subject, and white blood cells (WBC), neutrophils (NEUT), lymphocytes (LYMPH), red blood cells (RBC), RBC distribution width (RDW), platelets (PLT), mean PLT volume (MPV), and large PLT ratio (PLCR) were detected. The NEUT/LYMPH ratio (NLR) and PLT/LYMPH ratio (PLR) were calculated. t test was used to compare measurement data between groups. Multiple logistic regression analysis was performed for blood cells with P <0.05. The receiver operating characteristic curve (ROC curve) was used to calculate the area under the curve (AUC) and 95% confidence interval (95% CI) of each inflammatory indicator, and the optimal cutoff value was determined according to the Jorden index (sensitivity+specificity-1). Results:Compared with control group, WBC, NEUT, NLR, RDW, PLR were increased in NA-CRAO group, while RBC and LYMPH were decreased, with statistical significance ( t=9.68, 12.43, 9.47, 3.64, 5.54, 5.18, 0.46; P<0.001). There was no significant difference in PLT, MPV and PLCR between the two groups ( t=0.32, 1.56, 0.84; P>0.05). Multivariate logistic regression analysis showed that NLR was a possible risk factor for the occurrence of NA-CRAO (odds ratio=2.51, 95% CI 0.780-0.859, P=0.031). ROC curve analysis showed that the AUC predicted by NLR was 0.819, the optimal critical value was 3.05, and the sensitivity and specificity were 59.2% and 92.7%, respectively. Conclusions:In peripheral blood cells of NA-CRAO patients, NEUT is significantly increased and LYMPH is decreased. NLR is a possible risk factor for NA-CRAO.

Objective:To observe the clinical and imaging features of non-arteriotic central retinal artery occlusion (NA-CRAO) with internal boundary membrane detachment (ILMD), and to analyze its relationship with visual prognosis.Methods:A retrospective clinical study. A total of 88 patients with NA-CRAO hospitalized in Department of Ophtalmology, Xi'an People's Hospital (Xi'an Fourth Hospital) from January 2014 to June 2023 were included in the study. Best corrected visual acuity (BCVA), optical coherence tomography (OCT) and fluorescein fundus angiography (FFA) were performed. The BCVA test used the international standard visual acuity chart, which was statistically converted to the logarithm of the minimum angle of resolution (logMAR) visual acuity. OCT observed the presence of ILMD and the thickening of the inner retina and the disappearance of anatomical stratification. FFA recorded arm-retinal circulation time (A-Rct) and retinal arterion-distal filling time (FT), and observed ciliary retinal artery, fluorescein retrograde filling, cotton spots, luciferin nodal filling, macular non-perfusion, capillary fluorescein leakage, optic disc strong fluorescence, choroidal background weak fluorescence and other characteristics. According to whether there was ILMD, the patients were divided into ILMD group and non-ILMD group, with 44 cases and 44 eyes respectively. The two groups received the same treatment. The follow-up time was 30 days after treatment. The clinical, FFA characteristics and BCVA before and after treatment were compared between the two groups. t-test was used for comparison between groups. Results:In ILMD group and non-ILMD group, there were 43 cases of male and 1 case of female, respectively, and the proportion of male was significantly higher than that of female. Before and after treatment, the logMAR BCVA of ILMD group and non-ILMD group were 2.35±0.42, 2.01±0.46, 1.47±0.60, 0.77±0.49, respectively. There were significant differences in logMAR BCVA between the two groups before and after treatment ( t=8.025, 12.358; P<0.001). Before treatment, A-Rct and FT in ILMD group were longer than those in non-ILMD group, and the difference was statistically significant ( t=3.052, 3.385; P<0.05). After treatment, there was no significant difference ( t=1.040, 1.447; P >0.05). The proportion of ciliary retinal artery and cotton plaque in ILMD group was lower than that in non-ILMD group. There was no significant difference in ciliary retinal artery between the two groups ( χ2=-0.961, P>0.05), but there was a significant difference in cotton wool plaque between the two groups ( χ2=-3.364, P <0.05). Compared to the non-ILMD group, The proportion of retrograde fluorescein filling in retinal artery ( χ2=-2.846), segment filling ( χ2=-3.907), macular non-perfusion ( χ2=-6.656), capillary fluorescein leakage ( χ2=-4.367), optic disc strong fluorescence ( χ2=-3.525) and choroidal background weak fluorescence ( χ2=-2.276) increased, the difference was statistically significant ( P<0.05). Conclusions:In patients with NA-CRAO, compared with those without ILMD, those with ILMD have more severe retinal ischemia and worse BCVA before and after treatment. ILMD is one of the poor prognostic markers of NA-CRAO vision.

Rab27a gene can participate in the secretion, transportation and exocytosis of T lymphocytes, mast cells, neutrophils, pancreatic β cells and hematopoietic cells through its encoded proteins, and participate in the development of diseases such as albinism, cancer, diabetes, inflammation and thrombosis. In addition, the secretion regulation of Rab27a to exosomes has also received much attention. The current research status of the secretion of exosome and the genetically modified exosomes of Rab27a gene in the treatment of diseases such as inflammatory injury, thrombosis and tumor were reviewed.

Objective To study the effect of exosomes ( EXs) released from high expression of miR-132-3p mesenchymal stem cells (MSCs) on hypoxia/reoxygenation (H/R) injured endothelial cell function. Methods MSCs extracted from bone marrow of C57BL/6 mice were cultured primarily. MSCmiR-132-3p was obtained from MSCs infected with lentivirus loaded with miR-132-3p vector. At the same time,MSCNC was obtained by infecting MSCs with control lentivirus loaded with scramble sequence. EXs released from MSCNCand MSCmiR-132-3pwas isolated,and MSC-EXs and MSC-EXsmiR-132-3pwere obtained respectively. The obtained EXs and H/R damaged mouse brain microvascular endothelial cells (bend3) were co-cultured. According to culture conditions,the cells were divided into normal culture group (normal cell culture),H/R group (making a H/R model),MSC-EXs group (MSC-EXs co-culture),MSC-EXsmiR-132-3p group (MSC-EXsmiR-132-3pco-culture), and MSC-EXsmiR-132-3p+ LY294002 group ( before the cells and MSC-EXsmiR-132-3pwere co-cultured,treated by adding phosphatidyl alcohol 3 kinase [ PI3K] signaling pathway blocker LY294002 [20 μmol/L]). Quantitative real-time quantitative polymerase chain reaction was used to detect the expression of miR-132-3p in MSCs,MSC-EXs,and bend3 cells. Angiogenesis kit was used to detect angiogenic ability of bend3 cells,and 3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide (MTT) assay was used to detect the proliferative capacity of bend3 cells. Scratch test was used to detect the migration ability of bend3 cells. hochest33258 staining showed cell apoptosis. Western blot was used to detect the phosphorylation level of protein kinase B ( Akt) . Results Compared with the H/R group, the MSC-EXs treatment group significantly improved the angiogenesis,proliferation,migration abilities, and Akt phosphorylation level of bend 3 cell damage induced by H/R (The H/R group were 3 ± 1,0. 275 ± 0. 020,147 ± 8 μm,and 0. 89 ± 0. 12,respectively;the MSC-EXs treatment group were 8 ± 3,0. 358 ± 0. 030,218 ± 10 μm, and 1. 37 ± 0. 25 μm,respectively;all P<0. 01). Apoptosis was significantly reduced (47 ± 2% vs. 63 ± 2%,all P<0. 01). Compared with the MSC-EXs treatment group,the angiogenesis,proliferation,migration abilities,and Akt phosphorylation level of bend 3 cells in the MSC-EXsmiR-132-3ptreatment group were increased (14 ±3,0. 444 ± 0.050,357±10μm,and1.67±0.23,respectively,all P<0.01).Apoptosis was significantly reduced (34±1%,all P<0. 01) . Compared with the MSC-EXsmiR-132-3ptreatment group, cell proliferation, migration, angiogenesis abilities,and Akt phosphorylation level in the MSC-EXsmiR-132-3p+LY294002 group were significantly reduced (5 ± 2,0. 304 ± 0. 050,175 ± 8 μm and 0. 95 ± 0. 11,respectively,all P<0. 01). Conclusion MSC-EXs with high expression of miR-132-3p may improve many physiological functions of H/R-induced damaged cerebrovascular endothelial cells by activating PI3K/Akt signaling pathway.

Objective To investigate the expression levels of the CD14 mRNA and TLR4 mRNA in peripheral blood mononuclear cells from patients with Parkinson disease and to explore the clinical signicance. Methods For?ty-four patients with Parkinson disease and 37 healthy controls were recruited. We recorded age of onset, duration of ill?ness and sex of all recruited patients. PD patients were evaulated using the Hoehn-Yahr stages, Unified Parkinson Dis?ease Rating Scale (UPDRS) Ⅱand UPDRS Ⅲ,non-motor symptoms scale (NMSS) on“off”time. Reverse transcrip?tion-polymerase chain reaction was performed to determine the expression levels of CD14 mRNA and toll-like receptor 4 (TLR4) mRNA. Results The expression levels of CD14 mRNA(1.459±0.658)2-△△CT and TLR4 mRNA (1.408±0.698)2-△△CT was significantly up-regulated (P<0.05) in Parkinson disease compared with controls((1.162 ± 0.631)2-△△CT、(1.122 ± 0.557)2-△△CT). In addition, there was positive correlation between the expression levels of CD14 mRNA in Parkinson dis?ease patients with the Hoehn-Yahr stages. Meanwhile, there was no significant correlation between the expression levels of CD14 mRNA and TLR4 mRNA with other clinical scores. Conclusions There is positive correlation between the ex?pression levels of CD14 mRNA in Parkinson disease patients with the Hoehn-Yahr stages,indicating that CD14/TLR4 positive monocyte may be involved in the pathogenesis of the Parkinson disease.

Objective To investigate the sleep quality of patients with sudden hearing loss so as to provide basis for taking targeted nursing intervention. Methods A total of 98 patients with sudden hearing loss were investigated by Pittsburgh sleep quality index ( PSQI) and general data questionnaire from September 2014 to April 2015 in Internal Medicine Department of PLA General Hospital. Results The total score of PSQI in patients with sudden hearing loss was (6. 85 ± 3. 53) points including 62 cases (63. 3%) for ≤7 points and 36 cases (36. 7%) for >7 points. Study showed a correlation between sleep quality and age indicating that the younger of age, the worse of sleep quality. In addition, when sleep quality was poor, anxiety was high. Anxiety emotion existed in 14 patients with sudden hearing loss whose scores of PSQI were higher than 12 points ( u=6.623, P < 0. 01). Conclusions The incidence of sleep disorder is high in patients aged 20 to 40 with sudden hearing loss. The most serious problem is difficulty falling asleep. Medical staff members and patients should pay attention to population characteristics and influence of anxiety on sleep quality so as to help patients improve the cognition of disease, reduce anxiety mood and improve sleep quality by comprehensive health education.

OBJECTIVETo investigate the inhibitory effect of salinomycin on human breast cancer cells in vitro, and to explore the related molecular mechanism.

METHODSHuman breast cancer MDA-MB-231 cells were treated with salinomycin at different concentrations and at various time points. The effect of salinomycin on MDA-MB-231 cells proliferation was studied by CCK-8 method. The cell cycle status was examined by flow cytometry. RT-PCR and Western blot were used to detect the expression of Shh, Smo and Gli1 in the Hedgehog pathway at mRNA and protein levels.

RESULTSProliferation of MDA-MB-231 cells treated with salinomycin was markedly inhibited in a concentration and time dependent manner. Salinomycin at concentrations of 0, 0.4, 0.8 and 1.6 µmol/L inhibited the growth at the rates of 11.18%, 25.88%, 50.03%, 92.65%, respectively. Salinomycin prevented MDA-MB-231 cells from G1 into S phase. Salinomycin at concentrations of 0, 0.8 and 1.6 µmol/L resulted in S-phase percentage of 25.03%, 11.85% and 35.21%, respectively (P < 0.05). RT-PCR and Western blot showed that the expression of key elements Shh, Smo and Gli1 in the Hedgehog pathway was inhibited by salinomycin in a concentration dependent manner (P < 0.05).

CONCLUSIONSalinomycin prevents breast cancer cell transition from G1 to S phase through downregulation of the target genes of Hedgehog signaling pathway, leading to an effective inhibition of MDA-MB-231 cells.

Objective To investigate the inhibitory effect of salinomycin on human breast cancer cells in vitro, and to explore the related molecular mechanism.Methods Human breast cancer MDA-MB-231 cells were treated with salinomycin at different concentrations and at various time points.The effect of salinomycin on MDA-MB-231 cells proliferation was studied by CCK-8 method.The cell cycle status was examined by flow cytometry.RT-PCR and Western blot were used to detect the expression of Shh, Smo and Gli1 in the Hedgehog pathway at mRNA and protein levels.Results Proliferation of MDA-MB-231 cells treated with salinomycin was markedly inhibited in a concentration and time dependent manner.Salinomycin at concentrations of 0,0.4,0.8 and 1.6 μmol/L inhibited the growth at the rates of 11.18%,25.88%, 50.03%, 92.65%, respectively.Salinomycin prevented MDA-MB-231 cells from G1 into S phase.Salinomycin at concentrations of 0,0.8 and 1.6μmol/L resulted in S-phase percentage of 25.03%,11.85%and 35.21%, respectively ( P <0.05 ).RT-PCR and Western blot showed that the expression of key elements Shh, Smo and Gli1 in the Hedgehog pathway was inhibited by salinomycin in a concentration dependent manner( P<0.05) .Conclusion Salinomycin prevents breast cancer cell transition from G1 to S phase through downregulation of the target genes of Hedgehog signaling pathway, leading to an effective inhibition of MDA-MB-231 cells.