Objective To prepare a kind of P-selectin antibody loaded-targeted ultrasound contrast agent,and to investigate its ability of targeting in vitro.Methods P-selectin antibody loaded-targeted ultrasound contrast agent was prepared via an avidin-biotin bridge.Its basic property was determined.Human umbilical vein endothelial cells (HUVECs) were stimulated to express P-selectin by applying different doses of recombinant human interleukin-4 (rhIL-4) and histamine.The expression level of P-selectin was detected by immunofluorescence technique,and explored rhIL-4 optimal stimulation dose.The experiment group was divided into 3 groups,including targeted ultrasound contrast agent,isotype control ultrasound contrast agent and blank ultrasound contrast agent.Targeting abilities of 3 groups were observed by adhering to treated HUVECs and untreated HUVECs,respectively.Results P-selectin antibody loaded targeted ultrasound contrast agent was prepared successfully.The average diameter was (2.24 ± 0.71)μm.The average Zeta potential was (-2.75 ± 0.84)mV.The concentration was (3.0 ± 0.3) × 109/ml.The rate of antibody binding was as high as 99.80％.RhIL-4 and histamine could stimulate HUVECs to express P-selectin,and optimal stimulation dose was obtained.Targeting experiment showed that targeted ultrasound contrast agent could preferably adhere to HUVECs stimulated by optimal dose of rhIL-4 and histamine.Compared with other groups,there were significant differences.Conclusions P-selectin antibody loaded targeted ultrasound contrast agent was prepared successfully via an avidin-biotin bridge.The targeted ultrasound contrast agent could effectively adhere to HUVECs stimulated by rhIL-4 and histamine.

Triple-negative breast cancer (TNBC) is a heterogeneous disease.It has distinct risk factors,molecular biology features,clinical presentations and prognosis.TNBC recurrence is common after resection,and the survival rate is low and available treatment options are few after recurrence.To date,chemotherapy is the main treatment strategy for TNBC.There is a great need for new molecular predictive marks and drug targets for improving the efficacy of TNBC treatment.

Objective To explore the effectiveness of Failure Modes and Effects Analysis(FMEA)in emergency man-agement of sudden incidents involving outpatient and emergency patients in general hospitals,to provide references for the optimi-zation of emergency response process for such incidents.Methods Based on FMEA,we identified and evaluated risks in the e-mergency response procedures for sudden incidents involving outpatient and emergency patients in general hospitals.Potential fail-ure modes were analyzed to identify key risks with a Risk Priority Number(RPN)greater than 125.Continuous quality improve-ment measures were implemented to control these risks,and the effectiveness of these controls was evaluated using chi-square tests for statistical analysis.Results A total of 16 risk points in 4 major areas were identified.After implementing continuous quality improvement measures,the RPNs of these high-risk points decreased to below 125,effectively controlling the potential risks.This intervention significantly improved the utilization rate of emergency equipment,the timely reporting rate of sudden in-cidents,the timely feedback rate of emergency response,with statistically significant differences(P＜0.01).Conclusion The application of FMEA to outpatient and emergency management of sudden incidents helps optimize the emergency response process,thus enhancing the emergency response capability of general hospitals and ensuring effective handling of such incidents.

OBJECTIVETo observe the impact of concurrent administration of recombinant human p53 adenovirus (Ad-p53) with EGFR inhibitor gefitinib on breast cancer MDA-MB-468 cells.

METHODSMDA-MB-468 cells were treated with Ad-p53 and/or gefitinib. The effect of Ad-p53 and gefitinib on the growth of MDA-MB-468 cells was evaluated by MTT assay. Cell apoptosis was detected by flow cytometry. Western blot analysis was used to detect the alteration of p53,EGFR, phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway and apoptosis-related proteins. Ad-p53 combined with gefitinib was used in vivo to explore their effect on tumor xenograft in nude mice. Immunohistochemistry was used to detect the p53 expression in vivo.

RESULTSThe MTT assay showed a stronger inhibitory effect of gefitinib on MDA-MB-468 cells infected with Ad-p53 than on the control cells. Cell apoptosis assay revealed that the apoptosis rates of MDA-MB-468 cells in vehicle-treated group, Ad-p53 group, gefitinib group, and combination group were 8.5%, 17.4%, 20.5% and 32.6%, respectively. The apoptosis rate of MDA-MB-468 cells in the combination group was higher than that in other groups (P < 0.05, for all) . Western blot analysis revealed that the expression of p53 was significantly up-regulated in the presence of Ad-p53. The combination of Ad-p53 and gefitinib significantly down-regulated p-Akt (S473)(P < 0.01) and up-regulated caspase-9 and cleaved caspase-3 (P < 0.01 for both). Tumor inhibition rates (TIR) in the Ad-p53, gefitinib, and combination groups were 35.7%, 28.7% and 74.4%, respectively. Ad-p53 and gefitinib combination therapy significantly reduced the tumor burden in nude mice (P < 0.05 for all).Immunohistochemistry showed that after intratumoral administration of Ad-p53, wild-type p53 was increased (P < 0.01). p53 expressions in the vehicle-treated, Ad-p53, gefitinib and combination groups were 45.2%, 80.1%, 50.7% and 90.6%, respectively.

CONCLUSIONSWild-type p53 may reverse the sensitivity of MDA-MB-468 cells to gefitinib through down-regulation of the PI3K/Akt pathway. The apoptotic activity induced by this combination treatment might be regulated through caspase cascade.

Adenoviridae ; Animals ; Antineoplastic Agents ; pharmacology ; Apoptosis ; Breast Neoplasms ; Caspase 3 ; Caspase 9 ; Cell Line, Tumor ; Down-Regulation ; Genes, p53 ; Humans ; Mice ; Mice, Nude ; Phosphatidylinositol 3-Kinases ; Quinazolines ; pharmacology

Objective To investigate the effect of combined administration of autophagy inhibitor 3?methyladenine/bafilomycin A1 and EGFR inhibitor gefitinib on triple?negative breast cancer MDA?MB?468, MDA?MB?231 cells and estrogen receptor?positive MCF?7 cells. Methods All the cells were treated with 3?methyladenine/bafilomycin A1 and/or gefitinib. The effect of autophagy inhibitor and gefitinib on the cell growth was evaluated by MTT assay. Cell apoptosis was detected by flow cytometry. Western blot analysis was used to determine the alteration of autophagy?related protein ( such as LC3) and apoptosis?related proteins ( such as caspase?3 and caspase?9) . Results MTT assay showed that the IC50 in the GE+3?MA and GE+BAF groups were (4.1±0.2) μmol/L and (3.8±0.3) μmol/L, significantly lower than that of the gefitinib alone group [(7.0±0.2) μmol/L] in MDA?MB?468 cells (P<0.05). Similarly, the IC50 in the GE+3?MA and GE+BAF groups were (9.7±0.1) μmol/L and (7.7±0.2) μmol/L, significantly lower than that of the gefitinib alone group [(14.7±0.1) μmol/L]in MDA?MB231 cells (P<0.05). The flow cytometry assay revealed that the apoptosis rates of MDA?MB?468 cells in GE, GE+3?MA and GE+BAF groups were (12.43± 3.18)%, (23.37±2.71)% and (18.71±2.81)%, respectively. The apoptosis rates of MDA?MB?231 cells of the GE, GE+3?MA and GE+BAF groups were (12.15±1.82)%, (16.94±2.19)% and (33.83±5.92) %, significantly higher than that of the gefitinib alone group (All P<0.05). The apoptosis rates of the MCF?7 cells were not changed significantly among the three groups (P>0.05). Western blot data showed that the expression levels of LC3 and p?Akt were decreased in the combined groups than that of the gefitinib alone group, while the p?PTEN, caspase?3 and caspase?9 were increased. Conclusions Autophagy inhibitor may enhance the sensitivity to gefitinib in MDA?MB?468 and MDA?MB?231 cells by activation of the PTEN/P13K/Akt pathway. Apoptosis in MDA?MB?468 and MDA?MB?231 cells might be enhanced by the combination treatment through caspase cascade.

Objective To investigate the effect of combined administration of autophagy inhibitor 3?methyladenine/bafilomycin A1 and EGFR inhibitor gefitinib on triple?negative breast cancer MDA?MB?468, MDA?MB?231 cells and estrogen receptor?positive MCF?7 cells. Methods All the cells were treated with 3?methyladenine/bafilomycin A1 and/or gefitinib. The effect of autophagy inhibitor and gefitinib on the cell growth was evaluated by MTT assay. Cell apoptosis was detected by flow cytometry. Western blot analysis was used to determine the alteration of autophagy?related protein ( such as LC3) and apoptosis?related proteins ( such as caspase?3 and caspase?9) . Results MTT assay showed that the IC50 in the GE+3?MA and GE+BAF groups were (4.1±0.2) μmol/L and (3.8±0.3) μmol/L, significantly lower than that of the gefitinib alone group [(7.0±0.2) μmol/L] in MDA?MB?468 cells (P<0.05). Similarly, the IC50 in the GE+3?MA and GE+BAF groups were (9.7±0.1) μmol/L and (7.7±0.2) μmol/L, significantly lower than that of the gefitinib alone group [(14.7±0.1) μmol/L]in MDA?MB231 cells (P<0.05). The flow cytometry assay revealed that the apoptosis rates of MDA?MB?468 cells in GE, GE+3?MA and GE+BAF groups were (12.43± 3.18)%, (23.37±2.71)% and (18.71±2.81)%, respectively. The apoptosis rates of MDA?MB?231 cells of the GE, GE+3?MA and GE+BAF groups were (12.15±1.82)%, (16.94±2.19)% and (33.83±5.92) %, significantly higher than that of the gefitinib alone group (All P<0.05). The apoptosis rates of the MCF?7 cells were not changed significantly among the three groups (P>0.05). Western blot data showed that the expression levels of LC3 and p?Akt were decreased in the combined groups than that of the gefitinib alone group, while the p?PTEN, caspase?3 and caspase?9 were increased. Conclusions Autophagy inhibitor may enhance the sensitivity to gefitinib in MDA?MB?468 and MDA?MB?231 cells by activation of the PTEN/P13K/Akt pathway. Apoptosis in MDA?MB?468 and MDA?MB?231 cells might be enhanced by the combination treatment through caspase cascade.

Objective:To compare the application effect of blue dye single tracer and blue dye combined with nuclide double tracer in sentinel lymph node biopsy (SLNB) of breast cancer surgery.Methods:A total of 92 breast cancer patients in Shandong Cancer Hospital and Institute from November 2017 to October 2019 underwent methyleneblue dye combined with 99Tc m sulfur colloid nuclide double tracer in SLNB, while other 92 cases in Jining First People Hospital underwent blue dye single tracer. The number of SLN detection, detection rate, accuracy rate, sensitivity, and false negative rate of the two groups were compared. The impacts of age, menstruation, tumor location, tumor size, clinical stage, pathological type, and estrogen receptor (ER), progesterone receptor (PR), human epidermal receptor 2 (HER-2), molecular typing, dynamic enhanced magnetic resonance imaging (DCE-MRI)on the detection rate of SLN were analyzed. Results:The number of detection, detection rate, accuracy, sensitivity, and false negative rate of the blue dye single tracer group were 3.20±1.10, 90.22%, 93.48%, 95.24% and 4.76%, respectively; the double tracer group were 3.37±1.02, 92.39%, 95.65%, 95.65% and 4.35%, respectively, without significant difference (all P>0.05). In different age, menstrual condition, tumor location, clinical stage, pathological type, ER, PR, HER-2 expression and molecular typing, the detection rate of single tracer group and double tracer group had no significant difference (all P>0.05). However, in the tumor size of 2-5 cm and without DCE-MRI examination, the detection rate of single tracer group was significantly lower than that of double tracer group. Conclusion:The effect of blue dye single tracer in detecting SLN of breast cancer is equivalent to that of double tracer method, which is worthy of promotion and application in primary hospitals.

Objective:To investigate the clinical characteristics and offer diagnostic and therapeutic approaches for adult-onset idiopathic hypogonadotropic hypogonadism(AIHH).Methods:Clinical, laboratory, and imaging data, as well as follow-up information, of three male patients diagnosed with AIHH at the Department of Endocrinology and Metabolism of Nanfang Hospital, Southern Medical University, were systematically reviewed and analyzed.Results:All three patients were male, with a median age of 39 years(range, 22 to 40). Two patients reported symptoms of enlarged breasts and reduced sexual function, while one case solely reported a decline in sexual function. Physical examination showed that the median length of the penis was 6 cm(range, 5 to 6 cm), and the bilateral testicular volume was 7.96 mL(4.70-8.82 mL). Basal hormone levels at the time of initial visit to our hospital as follows: the median testosterone level was 0.32 ng/mL(0.24-2.96 ng/mL), median follicle stimulating hormone(FSH) level was 0.56 mIU/mL(0.1-0.75 mIU/mL), and the median luteinizing hormone(LH) level was 0.69 mIU/mL(0.1-1.03 mIU/mL). The levels of other hormones secreted by the anterior pituitary gland were normal. Hypothalamic-pituitary magnetic resonance imaging(MRI) showed that 1 patient had a pituitary microadenoma. Three patients were treated with pulsatile GnRH or gonadotropins, one of which had hypothalamic-pituitary-gonadal(HPG) axis function reversal after GnRH pulse pump therapy and lasted for 1 year, but then still had irreversible reduction.Conclusion:AIHH is marked by adult-onset disease and idiopathic hypogonadism. Enhancing fertility remains a critical requirement for these patients. Pulsatile GnRH treatment or gonadotropin therapy, as viable treatments, exhibit therapeutic effects, albeit with occasional fluctuations. Therefore, the emphasis lies in the timely consideration of fertility preservation.

Objective:To compare the application effect of blue dye single tracer and blue dye combined with nuclide double tracer in sentinel lymph node biopsy (SLNB) of breast cancer surgery.Methods:A total of 92 breast cancer patients in Shandong Cancer Hospital and Institute from November 2017 to October 2019 underwent methyleneblue dye combined with 99Tc m sulfur colloid nuclide double tracer in SLNB, while other 92 cases in Jining First People Hospital underwent blue dye single tracer. The number of SLN detection, detection rate, accuracy rate, sensitivity, and false negative rate of the two groups were compared. The impacts of age, menstruation, tumor location, tumor size, clinical stage, pathological type, and estrogen receptor (ER), progesterone receptor (PR), human epidermal receptor 2 (HER-2), molecular typing, dynamic enhanced magnetic resonance imaging (DCE-MRI)on the detection rate of SLN were analyzed. Results:The number of detection, detection rate, accuracy, sensitivity, and false negative rate of the blue dye single tracer group were 3.20±1.10, 90.22%, 93.48%, 95.24% and 4.76%, respectively; the double tracer group were 3.37±1.02, 92.39%, 95.65%, 95.65% and 4.35%, respectively, without significant difference (all P>0.05). In different age, menstrual condition, tumor location, clinical stage, pathological type, ER, PR, HER-2 expression and molecular typing, the detection rate of single tracer group and double tracer group had no significant difference (all P>0.05). However, in the tumor size of 2-5 cm and without DCE-MRI examination, the detection rate of single tracer group was significantly lower than that of double tracer group. Conclusion:The effect of blue dye single tracer in detecting SLN of breast cancer is equivalent to that of double tracer method, which is worthy of promotion and application in primary hospitals.

Complete androgen insensitivity syndrome(CAIS) is characterized by lack of androgen response in target organs due to androgen receptor dysfunction, resulting in feminized external genitalia. Individuals with CAIS are typically advised to live as females. This article reports a patient diagnosed with CAIS and gender dysphoria in adulthood. Following the removal of a left pelvic mass, pathology indicated cryptorchidism with a concurrent Leydig cell tumor. Genetic testing revealed a deletion mutation in exon 3 of androgen receptor gene. During follow-up, the patient underwent gender reassignment, transitioning socially from female to male. This case provides new insights into gender allocation for CAIS patients.