Objective To observe the osteoclast differentiation of Raw264.7 strain stably expressing enhanced green fluorescent protein(EGFP).Methods Raw264.7 cells were transfected with EGFP-Lifeact gene via retrovirus.The G3 cell clone was obtained by limited dilution technique which stably expressed EGFP under the fluorescence microscope.The morphology of G3 cells were observed.The effects of transfection on receptor activator of nuclear factor kappaB ligand(RANKL)-induced osteoclastogenesis and bone resorbing function of G3 cells were examined by tartrate resistant acid phosphatase(TRAP) staining,immunoblotting detection of cathepsin K and bone pit resorption assay.The real-time images of podosome dynamics were taken by laser confocal microscopy during osteoclast differentiation of G3 cells.Results The Raw264.7 cells were successfully transfected with EGFP-Lifeact gene.The G3-EGFP cloned strain which could stably express EGFP even after 20 passages was constructed.There was no significant difference in morphology between G3-EGFP and wild Raw264.7 cells.The fusion rates of the transfection group and of the wild control group were (35±5)％ and (39±5)％,respectively,which were not significantly different(P＞0.05).The semi-quantitative ratio of cathepsin K/β-actin in the wild control group and in the transfection group was 0.83±0.07 and 1.02±0.08(P＞0.05),respectively.Bone pit results showed that the total area of the bone resorption was respectively 272 252±36 193 and 262 408±23 243(P＞0.05) and the number of the bone pits was respectively 320 ± 51 and 339± 55(P＞0.05).The photos of laser confocal microscopy showed the constant cell-cell fusion during osteoclast differentiation of G3-EGFP cells.In addition,the dynamic self-organized podosome initially assembled podosome clusts,then dynamic rings,finally formed the characteristic podosome belt pattern in mature osteoclasts.Conclusions Enhanced green fluorescent protein high effectively expressed in Raw264.7.Biological character does not change after transfection.

Objective:To study the effect of surgical treatment on the psychological status of initial-diagnosed breast cancer patients.Methods:Using a prospective study method, 73 initial-diagnosed breast cancer patients who received surgical treatment in Beijing Bo′ai Hospital from March 2015 to June 2017 were selected. The self-rating anxiety scale (SAS), the self-rating depression scale (SDS) and the symptom checklist-90 (SCL-90) were used to evaluate the psychological health status before and after surgery by face-to-face conversation. The measurement data were described as mean±standard deviation( ± s), and the scores of SAS, SDS and SCL-90 before and after surgery were analyzed by paired t-test. Chi-square test was used to compare count data before and after surgery. Results:After surgical treatment, the anxiety and depression scores were 38.03±8.53 and 42.17±10.93, which were significantly lower than that in the pre-operation (46.47±11.47 and 51.01±14.22), the differences were statistically significant ( P<0.001). The scores of somatization, obsessive-compulsive symptoms, interpersonal sensitivity, anxiety, depression, hostility, phobia, paranoid and psychotic disorder of initial-diagnosed breast cancer patients in SCL-90 were 4.52±4.71, 6.15±5.49, 3.51±3.26, 4.52±4.44, 7.12±6.23, 1.55±2.03, 3.18±3.36, 0.99±1.36 and 2.85±3.59 after surgical treatment, which were significantly lower than that in the pre-operation (6.88±6.36, 10.08±6.77, 5.99±5.57, 10.03±8.00, 12.59±9.78, 4.12±5.25, 4.92±4.46, 2.59±3.60) and (5.84±5.43), the differences were statistically significant ( P=0.002 in somatization, others P<0.001). Conclusion:Surgical treatment can improve the psychological status of initial-diagnosed breast cancer patients, which has important significance for the psychological rehabilitation of patients who were initial-diagnosed breast cancer.

Objective:To investigate the relationship between the changes of inflammatory cytokine levels and prognosis of patients with critical coronavirus disease 2019 (COVID-19) undergoing invasive mechanical ventilation (IMV).Methods:A retrospective study was conducted. The clinical date of critical COVID-19 patients undergoing IMV who were hospitalized in Wuhan Union Hospital, Tongji Medical College of Huazhong University of Science and Technology from February 4th to March 25th in 2020 were collected. At the same time, the inflammatory cytokine levels including interleukins (IL-2, IL-4, IL-6, IL-10) and tumor necrosis factor-α (TNF-α) at 48 hours before IMV and 48 hours after IMV of all the patients, as well as the 48 hours after weaning or right before death were recorded. Multivariate unconditional Logistic regression analysis was used to screen the independent risk factors of death during hospitalization.Results:Among the 43 patients, 13 patients improved and 30 died. Compared with the survival group, the patients in the non-survival group were older (years old: 67.6±7.3 vs. 58.5±11.9, P < 0.05), with higher rates of hypertension, diabetes and coronary heart disease (53.3% vs. 15.4%, 63.3% vs. 23.1%, 26.7% vs. 0%, all P < 0.05), and the time from onset to admission to hospital, admission to ICU and IMV were longer (days: it was 9.17±5.00 vs. 5.07±2.49, 17.10±7.11 vs. 12.23±5.05, and 17.90±7.46 vs. 12.61±5.60, respectively, all P < 0.05). The IL-6 and TNF-α levels on 48 hours after IMV in the non-survival patients increased significantly as compared with those before 48 hours and the surviving patients. Especially, the IL-6 levels increased significantly as compared with those at 48 hours after IMV and 48 hours after weaning in the surviving patients [ng/L: 800.00 (194.25, 2 000.00) vs. 22.03 (6.66, 28.21), 3 204.00 (1 264.88, 5 000.00) vs. 5.00 (3.98, 12.27), both P < 0.01]. The IL-10 level before death in the non-survival patients increased significantly as compared with that at 48 hours after weaning in the surviving patients [ng/L: 55.89 (26.07, 100.14) vs. 3.53 (2.76, 12.36), P < 0.05]. There were no significant differences in the levels of IL-2 and IL-4 between the two groups at every time point. The variables of age, basic diseases, the IL-6 level after IMV were included in the multivariate unconditional Logistic regression analysis, which showed that age [odds ratio ( OR) = 0.821, 95% confidence interval (95% CI) was 0.695-0.968], hypertension ( OR = 0.027, 95% CI was 0.002-0.378), diabetes mellitus ( OR = 0.054, 95% CI was 0.005-0.611), coronary heart disease ( OR = 0.042, 95% CI was 0.002-0.968) and the IL-6 level after IMV ( OR = 0.902, 95% CI was 0.819-0.994) were independent risk factors for death during hospitalization in patients with critical COVID-19 undergoing IMV (all P < 0.05). Conclusions:The levels of inflammatory cytokine including IL-6, IL-10, and TNF-α increased significantly with aggravation in critical COVID-19 patients undergoing IMV, especially IL-6. IL-6 was an independent risk factor for death of critical COVID-19 patients undergoing IMV.

Objective To investigate the correlation between red blood cell distribution width(RDW),systemic immune-inflammation index(SII)and major depressive disorder(MDD).Methods The clinical data of 176 MDD patients hospitalized in the clinical psychology department of our hospital from 2020 to 2022 and 209 non-MDD comparators who were routinely examined in Hebei General Hospital were retrospectively analyzed.Blood analysis was conducted to obtain values of RDW,SII,and red blood cell distribution width/platelet ratio(RPR).The data was used to plot the receiver operator characteristic(ROC)curve to determine the optimal threshold and the area under the curve(AUC)for RDW to discriminate between patients and controls.Result Patients in the MDD group had significantly higher RDW[median and quartiles:13.20(12.70,13.98)vs.12.80(12.40,13.35)],and SII levels[median and quartiles 510.87(350.95,878.12)vs.405.33(313.74,539.92)]compared with those in non-MDD group(P<0.05).There was no significant difference in RPR between the two groups(P>0.05).Multifactorial logistic regression analysis showed that RDW was positively correlated with MDD after adjusting for confounders(OR=3.086,95% CI:1.926-4.944).ROC curve showed that the optimal threshold for RDW to differentiate the risk of developing MDD was 12.85,with an AUC of 0.647(95% CI:0.592-0.702;P<0.001).Conclusion Present study shows that high RDW is a risk factor for the occurrence of MDD and an important parameter for the risk of developing depression.

Objective: To explore the clinical study of different frequency repetitive transcranial magnetic stimulation in the treatment of senile schizophrenia. Methods: The 70 elderly schizophrenics admitted to Kangning Hospital Affiliated to Wenzhou Medical University from February 2018 to August 2019 were randomly divided into observation group (35 cases) and control group (35 cases). High frequency repetitive transcranial magnetic stimulation was used in the observation group and low frequency repetitive transcranial magnetic stimulation was used in the control group. Both groups were treated for 4 weeks. The changes of clinical global impression (CGI), positive and negative syndrome scale (PANSS), cognitive function rating scale for schizophrenia (SCoRS), scale of social-skills for psychiatric inpatients(SSPI) and serum sex hormone levels before and after treatment were compared between the two groups. Results: After treatment, the CGI and PANSS scores of the two groups were lower than those before treatment [observation group:(2.43 ± 0.37) scores vs. (5.61 ± 0.58) scores, (35.73 ± 6.57) scores vs. (79.95 ± 8.98) scores, t=27.346 and 23.512; control group: (3.37 ± 0.48) scores vs. (5.49 ± 0.62) scores, (40.91 ± 8.17) scores vs. (78.64 ± 9.63) scores, t=15.996 and 17.675, P<0.05]; after treatment, the CGI and PANSS scores of the observation group were lower than those of the control group (t=9.176, 2.923, P<0.05). After treatment, the SCoRS scores of the two groups was lower than those before treatment (observation group: t=12.822, control group: t=7.402, P<0.05); after treatment, the SCoRS score of the observation group was lower than that of the control group [(27.92 ± 5.41) scores vs. (34.25 ± 6.17) scores] (t=4.564, P<0.05). The SSPI scores of the two groups increased after treatment (observation group: t=9.252; control group: t=3.671, P<0.05); after treatment, the SSPI scores of the observation group were higher than those of the control group [(35.74 ± 3.27) scores vs. (30.85 ± 2.28) scores](t=7.257, P<0.05). After treatment, the serum testosterone and prolactin levels in the two groups were higher than those before treatment (observation group: t=13.853, 15.198; control group: t=10.075 and 9.225, P<0.05); after treatment, the serum testosterone and prolactin levels in the observation group were higher than those in the control group (t=5.693 and 7.202, P<0.05). Conclusions High frequency repetitive transcranial magnetic stimulation is superior to low frequency repetitive transcranial magnetic stimulation in the treatment of senile schizophrenia, and the improvement of sex hormone level is superior to low frequency repetitive transcranial magnetic stimulation, and the improvement of cognitive and social functions is superior to low frequency repetitive transcranial magnetic stimulation.