Objective To investigate the effects of interferon-gamma (IFN-γ) on migration,invasion and vasculogenic mimicry (VM) formation of human melanoma cell line MUM-2B. Methods MUM-2B cells were divided into three groups, control group (10%FBS in DMEM), treatment group1 (10μg/L IFN-γ) and treatment group2 (100μg/L IFN-γ). Different concentrations of IFN-γ were added in the culture medium of MUM-2B cells. Wound-healing assay and matrigel invasion assay were performed to examine the migration and invasion ability of MUM-2B cells. Three-D culture was used to observe the VM formation. The expression of vascular endothelial growth factor (VEGF) of MUM-2B cells was detected by Western blot assay. Results The result of wound-healing assay showed that the migration distance of cells was decreased in treatment groups compared with that of control group. The migration distance of cells was decreased in treatment group 2 compared with that of treatment group 1(P＜0.05). The result of matrigel invasion assay showed that the number of invaded cells was decreased in treatment groups compared with that of control group, and which was significantly decreased in treatment group2 than that of treatment group1 (P＜0.05). The result of 3-D culture showed that cells in control group can form typical VM tube-like structures, whereas cells in treatment groups cannot. Western blot assay showed that the expression of VEGF protein was significantly decreased in treatment groups compared with that of control group, and the expression of VEGF protein was significantly decreased in treatment group2 than that of treatment group 1(P＜0.05). Conclusion These data suggest that IFN-γinhibits migration and invasion of MUM-2B cells, and inhibits VM formation by down regulating VEGF expression in vitro.

Objective To compare the timeliness, success and mortality rates between the modified carotid artery puncture method ( MCAPM) and standard suture method ( SSM) in the establishment of rat model of a middle cerebral ar?tery occlusion ( MCAO) . Methods Thirty?two male rats were randomly and equally assigned into MCAPM group and SSM group. The MCAO models were established by inserting a thread into the common carotid artery ( CCA) . 24 h after modeling, the rats of the two groups were evaluated with Bederson neurological scores, and the modeling success rate and mortality rate were analyzed. Results The suture insertion times, success rates and mortality rates of the MCAPM vs. SSM groups were (82?3 ±17?4) s versus (164?6 ± 22?0) s (P<0?01), 87?5% versus 68?75% (P>0?05), and 6?25% versus 18?75% (P>0?05). Conclusions MCAPM can be used to establish the rat model of MCAO due to its simplicity, mild wound and feasibility.

Objective To assess the role of the ferroptosis-associated gene GLS2 in the prognosis of pan-cancer and immunity using bioinformatics methods. Methods GLS2 expression levels in pan-cancer were profiled using publicly available databases: The Cancer Genome Atlas, GTEx, Cancer Cell Line Encyclopedia and the International Cancer Genome Consortium. The aim was to explore the role of GLS2 gene expression, gene variation survival analysis, immune infiltration, immune checkpoint related genes, TMB, and MSI in different tumors. Results Difference in GLS2 expression levels between cancer and paraneoplastic tissues was statistically significant in most cancer types, and the expression level was correlated with survival in these cancer types. A positive correlation was found between GLS2 expression and immune cell infiltration in multiple cancer types, and GLS2 expression level was positively correlated with TMB, MSI, and methylation, and its expression is an indicator for potential therapeutic response. Conclusion Pan-cancer analysis shows that the ferroptosis-related gene GLS2 can be used as a diagnostic and prognostic marker of clear cell carcinoma of kidney, adrenocortical carcinoma, lung adenocarcinoma, and pancreatic cancer.

Objective:To observe the effect of electroacupuncture(EA)combined with repetitive transcranial magnetic stimulation(rTMS)on limb dysfunction in ischemic stroke patients. Methods:A total of 63 stroke patients were divided into an observation group and a control group using the random number table method.Thirty-one patients in the control group were treated with routine Western medicine combined with rTMS;32 patients in the observation group were treated with EA in addition to the intervention in the control group.The duration of treatment was 3 months.The National Institutes of Health stroke scale(NIHSS),the Fugl-Meyer assessment(FMA),the modified Barthel index(MBI),and the motor evoked potential(MEP)latency of transcranial magnetic stimulation were observed before and after treatment in both groups. Results:Two cases withdrew from the observation group and 1 case withdrew from the control group.After treatment,the NIHSS score in both groups was lower than that before treatment,the FMA and MBI scores were higher than those before treatment,and the latency period of MEP was shorter than that before treatment,and the differences were statistically significant(P<0.05).After treatment,the NIHSS,FMA,and MBI scores and MEP latency period of the observation group improved more than those of the control group,and the differences between the groups were statistically significant(P<0.05). Conclusion:EA combined with rTMS can improve the motor function of limbs in ischemic stroke patients and improve their self-care ability.The mechanism may be related to increasing the excitability of the motor cortex and improving the electrophysiological function of the central nervous system.

Objective:To evaluate the cognitive function of elderly inpatients in Qinghai and analyze the influencing factors in this prospective study.Methods:A total of 1077 elderly inpatients were enrolled for general information investigation, including the Mini Nutritional Assessment-Short form(MNA-SF), Geriatric Depression Scale 15(GDS-15), FRAIL Scale, activities of daily living(ADL), instrumental ability of daily living(IADL)and the Mini-Mental State Examination(MMSE).Results:The 1077 elderly inpatients being subjected in this study, included 470 in the cognitive impairment(CI)group and 607 in the normal group.The prevalence of CI in hospitalized elderly patients was 43.64%(470/1077). The results of univariate analysis showed that as compared with the normal group, the proportion of cognitive impairment was higher in female, elder patients, and boiling tea-drinker, otherwise the proportion of CI was lower in patients living with the spouse, normal sleep, often eating beef and mutton( P<0.01 or P<0.05); FRAIL Scale, ADL total score, and IADL total score were higher in the normal group than in the CI group( P<0.01); the elderly depression score and frailty score were higher in the CI group than in the normal group( P<0.01). The results of logistic regression analysis showed that the risk of CI was higher in female and people frequently drinking boiling tea than in male and people not frequently drinking boiling tea( OR=0.740, 1.211, both P<0.05), and that frequent consumption of beef and mutton, and reasonable levels of nutrition, ADL, and IADL were protective factors for CI in elderly hospitalized patients( OR=0.780, 0.938, 0.956, 0.895, all P<0.05). Conclusions:The incidence of cognitive impairment in hospitalized elderly patients in Qinghai is relatively high.Female and often drinking boiling tea may increase the risk of cognitive impairment.Regularly eating beef and mutton, reasonable levels of nutrition, maintaining good daily life ability can delay cognitive impairment.

Objective To investigate the training demands for nursing knowledge related to plateau disaster and skill training among nursing staffs,in order to provide references for further training.Methods By cluster sampling method and self-designed questionnaire,the needs of nurses for nursing knowledge and skills including demands in the stage of preparation and first aid,demands in the stage of post-disaster and plateau disaster were investigated among 635 nurses from 10 hospitals in Tibet plateau area.Results The total scores of needs for nursing knowledge and skills related to plateau disaster was around (3.85±1.09). The scores of demands for nursing knowledge and skills in nurses with different ranks of hospital,nursing ages,professional titles,educational backgrounds and duties,were significant different (χ2=290.32,421.71, 462.92,236.71,489.43;P<0.01).Conclusions Nursing staffs have great demands for participating in training class related to nursing knowledge and skills of plateau disaster,in order to improve their nursing ability in plateau disaster.

Objective:To explore the local prevalence of early cognitive dysfunction after a stroke in some hospital of Qinghai province, analyze its risk factors and construct a risk factor model.Methods:A prospective cross-sectional survey was conducted among 854 stroke patients at 3 hospitals in Qinghai Province. The survey solicited demographic data, clinical data, and information about the respondents′ diet and leisure activities. Within 14 days after stroke onset, all of the participants′ cognitive performance was quantified using the Mini-Mental State Examination (MMSE). Those with scores ≤27 formed the cognitive impairment group ( n=569), while the rest were the normal cognition group ( n=285). Through univariate analysis and multivariate logistic regression analysis, the relationship between each factor and the outcome was clarified, and a prediction model was established in the form of a regression equation. The model′s receiver operating characteristics (ROC) curve was computed. Results:The incidence of acute cognitive impairment after stroke was 66.6%. Univariate analysis showed that there were significant differences between the two group in terms of age, sex, ethnicity, education, occupation, altitude, living with a spouse and social support. Stroke type, hemiplegia, a history of hypertension, and triglyceride and D-2 polymer levels were significant predictors, as were ADL score, leisure activity level, and weekly consumption of fruits, nuts and salt. The multivariate logistic regression analysis showed that age, being female, hemorrhagic stroke, hypertension, and living at high altitude were independent risk factors for cognitive impairment at the acute stage of stroke. However, education, occupation, leisure activity and consumption of triglycerides and nuts were independent protective factors. The prediction model showed an area under the ROC curve of 0.832, with sensitivity of 0.698 and specificity of 0.814.Conclusions:The prevalence of cognitive dysfunction in the acute stage of stroke is high in some hospital of Qinghai province. Age, being female, hemorrhagic stroke, hypertension and high altitude living are independent risk factors for stroke, while education, an occupation, leisure activity and triglyceride and nut consumption are protective factors. The risk factor model established in this study has good predictive ability with this population, with whom it can be used to formulate interventions to improve the life quality of stroke survivors.

Ferroptosis is a novel regulatory cell death characterized by iron dependence and mainly caused by the accumulation of lipid peroxides and reactive oxygen species in the cell. This process plays an important role in the development of many malignancies, and has been extensively studied in lung cancer, especially in antitumor therapy. In recent years, the role of ferroptosis in tumor immunotherapy has been gradually explored. Studies showed that targeting ferroptosis can improve the therapeutic efficacy of antitumor immunotherapy. In addition, immunotherapy and ferroptosis can work synergistically to enhance the effectiveness of antitumor therapy, suggesting a potential relationship between ferroptosis and immunotherapy and the possible reversal of immune drug resistance. This study aims to elucidate the characteristics of ferroptosis, and the role and potential clinical applications of ferroptosis in the antitumor immunotherapy of advanced non-small cell lung cancer. We also explore the role of some nanomaterials that target the onset of tumor ferroptosis in facilitating immunotherapy.

Objective This study aimed to explore the relationship between the spontaneous changes of mesiodistal angulation of lower second molars(L2M)and individual clinical characteristics in patients with early permanent dentition correction.Methods A total of 44 patients with a total of 88 mandibular second molars were included in this study after screening.Based on the frequency distribution histogram of the initial angle of inclina-tion(L2M initial angle)of the L2M crown before treatment,it was divided into three groups:Group A:small angle group,L2M initial angle is 0°～17.5°;Group B:medium angle group,L2M initial angle is 17.5°～42.5°;Group C:large angle group,L2M initial angle is 42.5°～60°.According to whether L2M has erupted and exposed most of the crown,it was divided into erupted group and unerupted group.According to the mandibular plane angle,it can be divided into high angle,low angle,and average angle.CBCT reconstructed panoramic images were used to measure the mesiodistal inclination of L2M and analyze the relationship between each group and the change in mesiodistal inclination angles of L2M.Results After treatment,L2M mesiodistal angulation in the small angle,large angle,non-eruption,and high angle groups became more significant.The angle in the middle angle group was significantly smaller.The L2M mesiodistal angulation in the eruption and average angle group did not change significantly.The angles of ANB,Y-axis,and mandibular plane have no correlation with the initial angle of L2M.Conclusion The L2M in patients with large and high angles is more likely to topple during a correc-tion.During the correction,the L2M tends to approach the normal axial inclination.

BACKGROUND: The aim of this study is to investigate the changes of peripheral blood lymphocyte subsets before and after treatment with pembrolizumab for non-small cell lung cancer and its clinical significance. METHODS: A total of 32 patients with non-small cell lung cancer who received pembrolizumab treatment in The Affiliated Hospital of Qingdao University and Weifang People's Hospital of Shandong Province from January 2015 to December 2020 were selected as the observation group, and 30 healthy people during the same period were selected as the control group. Before treatment and in cycles 1, 2 and 4 after treatment, fluid cytometry was used to detect changes in the levels of lymphocyte subsets in the peripheral blood of patients. RESULTS: The CD3⁺, CD4⁺, CD4⁺/CD8⁺ indexes of patients with non-small cell lung cancer before the treatment were significantly lower than those in the control group (P<0.05), and the CD8⁺ level was significantly increased (P<0.05); After 1 cycle of pembrolizumab treatment, there was no significant difference in the changes of lymphocyte subsets compared with before immunotherapy; After 2 cycles of the treatment, the CD3⁺, CD4⁺, CD4⁺/CD8⁺ values were higher than before the treatment (P>0.05), and the CD8⁺ index was slightly lower than before the treatment (P<0.05); After the fourth cycle of treatment, the CD3⁺, CD4⁺, CD4⁺/CD8⁺ values were significantly improved compared to before the treatment (P<0.05), and the CD8⁺ index was significantly lower than before the treatment (P<0.05); In the treatment process of patients with stable disease (SD)/partial response (PR), the CD3⁺, CD4⁺, CD4⁺/CD8⁺ values of the fourth cycles were higher than before the treatment (P<0.05), and the CD8⁺ index was lower than before the treatment (P<0.05); During the treatment of progressive disease (PD) patients, the changes of lymphocyte subsets in the fourth cycles were not significantly different from those before the treatment (P>0.05). At the same time, this article shows through analysis that the expression of programmed cell death ligand 1 (PD-L1) and pathological types have no obvious influence on the effect of immunotherapy. Multi-factor analysis shows that it is more meaningful to observe the changes of CD3⁺, CD4⁺ and CD8⁺ at the same time to predict the effect of immunotherapy. CONCLUSIONS Pembrolizumab can regulate the changes of T lymphocyte subsets in patients with non-small cell lung cancer, improve the immune status of the patients, and there is no obvious adverse reaction. At the same time, monitoring the changes of lymphocyte subsets during treatment can predict the effect of immunotherapy.