OBJECTIVES: Hypoxia can alter the oral bioavailability of drugs, including various substrates (drugs) of P-glycoprotein (P-gp), suggesting that hypoxia may affect the function of P-gp in intestinal epithelial cells. Currently, Caco-2 monolayer model is the classic model for studying the function of intestinal epithelial P-gp. This study combines the Caco-2 monolayer model with hypoxia to investigate the effects of hypoxia on the expression and function of P-gp in Caco-2 cells, which helps to elucidate the mechanism of changes in drug transport on intestinal epithelial cells in high-altitude hypoxia environment. METHODS: Normally cultured Caco-2 cells were cultured in 1% oxygen concentration for 24, 48, and 72 h, respectively. After the extraction of the membrane proteins, the levels of P-gp were measured by Western blotting. The hypoxia time, with the most significant change of P-gp expression, was selected as the subsequent study condition. After culturing Caco-2 cells in transwell cells for 21 days and establishing a Caco-2 monolayer model, they were divided into a normoxic control group and a hypoxic group. The normoxic control group was continuously cultured in normal condition for 72 h, while the hypoxic group was incubated for 72 h in 1% oxygen concentration. The integrity and polarability of Caco-2 cells monolayer were evaluated by transepithelial electrical resistance (TEER), apparent permeability (Papp) of lucifer yellow, the activity of alkaline phosphatase (AKP), and microvilli morphology and tight junction structure under transmission electron microscope. Then, the Papp of rhodamine 123 (Rh123), a kind of P-gp specific substrate, was detected and the efflux rate was calculated. The Caco-2 cell monolayer, culturing at plastic flasks, was incubated for 72 h in 1% oxygen concentration, the expression level of P-gp was detected. RESULTS: P-gp was decreased in Caco-2 cells with 1% oxygen concentration, especially the duration of 72 h (P<0.01). In hypoxic group, the TEER of monolayer was more than 400 Ω·cm², the Papp of lucifer yellow was less than 5×10^-7 cm/s, and the ratio of AKP activity between apical side and basal side was greater than 3. The establishment of Caco-2 monolayer model was successful, and hypoxia treatment did not affect the integrity and polarization state of the model. Compared with the normoxic control group, the efflux rate of Rh123 was significantly reduced in Caco-2 cell monolayer of the hypoxic group (P<0.01). Hypoxia reduced the expression of P-gp in Caco-2 cell monolayer (P<0.01). CONCLUSIONS Hypoxia inhibits P-gp function in Caco-2 cells, which may be related to the decreased P-gp level.
Humans ; ATP Binding Cassette Transporter, Subfamily B, Member 1 ; Caco-2 Cells ; ATP Binding Cassette Transporter, Subfamily B ; Hypoxia ; Oxygen

OBJECTIVE To optimize the pr eparation technology of the baicalin lipid nano foam aerosol （BC-LN-FA）. METHODS Baicalin lipid nanoparticle （BC-LN）and BC-LN-FA were prepared by the thin film dispersion method and homogeneous emulsification method ，respectively，using baicalin （BC） as the model drug. The preparation technology was optimized by Box-Behnken design-response surface methodology using particle size and encapsulation efficiency （EE）as indexes ，with dosage ， emulsifier dosage ，co-emulsifier dosage and homogenization time as factors. The morphology ，particle size ，polymerdispersity index（PDI），EE，the viscosity ，the foam dissolution rate and in vitro transdermal release of BC-LN-FA were characterized. RESULTS The optimal technology included 25 mg BC ，40 mg emulsifier （mass ratio of stearic acid-soybean lecithin-glycerol was 1∶1∶1），30 mg co-emulsifier （mass ratio of octadecanol-lactic acid was 1∶1），homogenization time of 20 min. Results of 3 times of validation tests showed that particle size of prepared BC-LN-FA was （151.70±2.40）nm，EE was （68.62±1.16）％；the deviation of them from the predicted value （particle size of 150.80 nm，EE of 67.02％）were 0.60％ and 2.39％ respectively. The BC-LN-FA prepared by the optimal process was light yellow opalescence ，uniform in particle size and round-like in shape. The viscosity，the foam dissolution rate ，the content of BC and PDI were （122.92±5.09）mPa·s，（65.32±3.22）％，（7.01±0.12）％ and（0.199±0.006），respectively. At 48 h，the cumulative release rates of BC-LN-FA in phosphate buffer saline （PBS）at pH 7.4， 6.8，5.0 were（54.12±2.69）％，（57.85±4.25）％ and（59.47±1.83）％，respectively；those of free BC in PBS at pH 7.4 was only （15.04±1.43）％. CONCLUSIONS The optimized technology is stable and feasible. Prepared BC-LN-FA has a uniform particle size，high digestion rate and certain viscosity.

Objective:To understand the current status of nurses' awareness and needs for the Employee Assistance Program (EAP), as to provide basis for the hospital to formulate EAP.Methods:Using the purpose sampling method, 352 ICU nurses from the Nanjing Drum Tower Hospital Affiliated of Nanjing University Medical School were selected as the research subjects from December 2019 to January 2020. Finally, 350 valid questionnaires were obtained. Use the EAP demand questionnaire to investigate the ICU nurses' awareness and demand for EAP.Results:Among the 350 nurses in the intensive unit, 214 cases (61.14%) had never understood EAP, only 8 (2.29%) had understood and experienced the EAP, and 332 cases (94.86%) believed that the hospital′s implementation of EAP was a benefit provided to nurses. The top two service requirements were: "Relief of work pressure" and "mental health consultation", and hope to receive mixed-mode EAP services; in ICU and CCU, nurses hope EAP can help resolve work-family conflicts; in EICU, they need to provide Specific job skills training services.Conclusions:ICU nurses have insufficient cognition of EAP and urgently need to obtain EAP implementation knowledge; at the same time, it is necessary to formulate an implementation plan that meets ICU nurses′ needs for EAP, that is, to relieve work pressure and carry out psychological counseling to ensure that EAP is launch successfully in ICU nurses.

Objective:To explore the effect and mechanism of fentanyl on promoting the stemness of breast cancer cell.Methods:From January 2018 to October 2019, human breast cancer cell line BT549 was used as the in vitro research object. Breast cancer cell BT549 was pretreated with 0.01 and 0.10 μmol/L fentanyl. Sphere formation assay and colony formation assay were performed to investigate the role of fentanyl on breast cancer cell stemness. Fluorescent quantitative polymerase chain reaction (FQ-PCR) was used to detect the mRNA level of stemness-related transcription factors gender-determining area Y box protein 2 (Sox2), octamer binding transcription factor 4 (Oct4) and Nanog. Western blotting assay was performed to determine the level of Wnt3a/β-catenin pathway-related markers Wnt3a, phosphorylated glycogen synthase kinase-3β (p-GSK-3β), glycogenase kinase-3β (GSK-3β) and β-catenin. After down-regulating Wnt3a, western blotting assay and sphere formation assay were performed.Results:The sphere diameter, colony formation rate and the expression of Sox2 mRNA, Oct4 mRNA, Nanog mRNA, Wnt3a, p-GSK-3β, GSK-3β and β-catenin in 0.01 and 0.10 μmol/L fentanyl-treated breast cancer cell were significant higher than those in blank control: (131.22 ± 1.06) and (636.37 ± 0.02) μm vs. (72.68 ± 0.13) μm, (41.33 ± 0.03)% and (60.58 ± 1.08)% vs. (20.93 ± 0.15)%, 2.25 ± 0.20 and 3.82 ± 0.84 vs. 1.00, 1.87 ± 1.06 and 3.35 ± 0.04 vs. 1.00, 2.85 ± 0.03 and 4.36 ± 0.50 vs. 1.00, 1.82 ± 0.03 and 2.57 ± 0.42 vs. 1.00, 2.04 ± 0.13 and 2.81 ± 0.05 vs. 1.00, 1.62 ± 0.17 and 2.93 ± 0.06 vs. 1.00, 2.15 ± 0.02 and 3.54 ± 0.21 vs. 1.00, the indexes in 0.10 μmol/L fentanyl-treated breast cancer cell were significantly higher than those in 0.01 μmol/L fentanyl-treated breast cancer cell, and there were statistical differences ( P<0.01). After down-regulating Wnt3a, the expressions of p-GSK-3β, GSK-3β, β-catenin, Sox2, Oct4 and sphere diameter were significantly lower than those in blank control: 0.12 ± 0.05 vs. 1.00, 0.53 ± 0.06 vs. 1.00 and 0.24 ± 0.21 vs. 1.00, 0.28 ± 0.10 vs. 1.00 and 0.06 ± 0.01 vs. 1.00, (18.14 ± 0.30) μm vs. (74.32 ± 0.12) μm, and there were statistical differences ( P<0.01). Conclusions:Fentanyl promotes breast cancer cell stemness by Wnt3a/β-catenin signaling pathway.

Objective: To investigate the role of H3K27M mutant in spinal cord glioma, specifically the correlation between H3K27M mutation and histological grade or prognosis. Methods: Twenty-four cases of paraffin-embedded spinal cord glioma tissues and clinical data were collected from November 2014 to August 2016 at the Beijing Tsinghua Changgung Hospital. There were 13 males and 11 females, and the age ranged from 3 to 66 years. All the cases were reviewed histologically, and immunohistochemical H3K27M staining and H3 gene detection were performed. The correlation between H3 gene mutation and histological grading and prognosis of spinal cord gliomas were investigated and relevant literature reviewed. Results: Eleven of 24 cases showed H3K27M gene mutation, and was concordant with the result of immunohistochemistry. Gliomas in the mutant group were all high-grade gliomas with mean patients′ age of (30.0±11.5) years, and a male to female ratio of 7:4. Thirteen cases were wild-type, and these included four high-grade gliomas, with mean patients′ age (31.3±22.4) years, and a male to female ratio of 6∶7. The tumors in the mutant group were mainly located in cervical 4-7 and thoracic 11-12 segments, respectively, and the incidence of tumors in the lower thoracic segments (thoracic 11-12) was higher than that in the wild type group. Outcome data were available for all patients. The median survival of mutant group was 19.5 months, but most patients in the wild-type group were alive at the end of the follow-up period. Conclusion Gliomas of spinal cord with H3K27M mutation are high-grade and the prognosis of patients is poor.

Objective To investigate the role of H3K27M mutant in spinal cord glioma, specifically the correlation between H3K27M mutation and histological grade or prognosis. Methods Twenty?four cases of paraffin?embedded spinal cord glioma tissues and clinical data were collected from November 2014 to August 2016 at the Beijing Tsinghua Changgung Hospital. There were 13 males and 11 females, and the age ranged from 3 to 66 years. All the cases were reviewed histologically, and immunohistochemical H3K27M staining and H3 gene detection were performed. The correlation between H3 gene mutation and histological grading and prognosis of spinal cord gliomas were investigated and relevant literature reviewed. Results Eleven of 24 cases showed H3K27M gene mutation, and was concordant with the result of immunohistochemistry. Gliomas in the mutant group were all high?grade gliomas with mean patients′age of (30.0±11.5) years, and a male to female ratio of 7:4. Thirteen cases were wild?type, and these included four high?grade gliomas, with mean patients′age (31.3±22.4) years, and a male to female ratio of 6∶7. The tumors in the mutant group were mainly located in cervical 4-7 and thoracic 11-12 segments, respectively, and the incidence of tumors in the lower thoracic segments (thoracic 11?12) was higher than that in the wild type group. Outcome data were available for all patients. The median survival of mutant group was 19.5 months, but most patients in the wild?type group were alive at the end of the follow?up period. Conclusion Gliomas of spinal cord with H3K27M mutation are high?grade and the prognosis of patients is poor.

Objective To observe the expression of PAX-2 and PTEN in different types of endometrial lesions, and to study their relationship with endometrial intraepithelial neoplasia (EIN). Methods 60 cases of endometrial hyperplasic lesions and 70 cases of endometrial carcinoma were enrolled. All cases were reclassified by using the diagnostic criteria of EIN, and PAX-2 and PTEN were stained to compare the difference among them. Results The deletion rates of PAX-2 in benign hyperplasia, EIN and endometrial carcinoma were 39.5 % (15/38), 72.7 % (16/22) and 78.6 % (55/70), respectively, and there was a statistical difference (χ2= 21.664, P= 0.000). The deletion rates of PTEN in benign hyperplasia, EIN and endometrial carcinoma were 47.4%(18/38), 54.5%(12/22) and 75.7%(53/70), respectively, and there was no statistical difference (χ2=2.878, P=0.411). Conclusion The staining of PAX-2 could be considered as a reliable adjuvant diagnostic method in the diagnostic criteria of EIN, however, the loss of PTEN just should be regarded as a suggestion of EIN, not a confirmed diagnostic basis.

Obgective To observe the effect of hydrogen sulfide (H2S) on vasorelaxation and expression of guanosine 3',5'-cyclic phosphate (cGMP) and activity of cyclic nucleotide phosphodiesterase (PDE) in vascular tissue.Methods H2S donor was provided by sodium bisulfide sodium hydrosulfide.The isolated perfused rat thoracic aorta rings were used to test the relaxation responses to H2S,which recorded by Power Lab system,and the enzyme linked immuno assay was used to detect intracellular cGMP.The activity of PDE was evaluated by using cyclic nucleotide PDE assay kit.Results (1) H2S relaxed the thoracic aorta rings,and the half maximal effective concentration (EC50) for the H2 S relaxation curve,represented by the corresponding concentration of H2 S that achieved 50％ of the maximum relaxation effect,was (1.79 ± 0.31) × 10-5 mol/L.(2)The cGMP content in vascular tissue increased from (22.29 ± 1.59) pmol/L to(41.45 ± 7.49) pmol/L and (31.35 ± 2.56) pmol/L after incubation with 50 μmol/L and 300 μmol/L H2 S,respectively (t =-3.09,t =-2.88;all P ＜ 0.05,n =7-8).(3) cGMP could be lysed into 5'-guanylicacid(5'-GMP) by PDE,which was an important pathway for cGMP degradation.This study showed that PDE activity was decreased in vascular tissue,the 5'-GMP decreased from (0.52 ±0.06) mol/L to (0.25 ±0.06) mol/Land (0.27 ±0.07) mol/L after incubation with 50 μmol/L and 300 μ mol/L H2S,respectively (t =3.21,t =2.58;all P ＜ 0.05,n =7-8).Conclusion The vasorelaxant effects of H2 S might be related to the inhibited activity of PDE and elevated content of cGMP.

ABSTRACT:Objective To explore the application value of psychological nursing in treatment of patients with malignant tumor chemotherapy.Methods A total of 105 patients with malignant tumor chemotherapy were randomly divided into intervention group (52 cases)and control group (53 cases).Control group was treated with routine nursing,on this basis intervention group was added with psychological nursing,including evaluation of psychological condition,establishment of favorable nurse-patient relationship,regular communication,optimization of patients’social sup-port network and relaxation training,etc.Results After nursing,self-rating anxiety scale (SAS) and self-rating depression scale (SDS)scores of both groups decreased significantly,which were significantly lower in intervention group than control group (P <0.01).After nursing,there were no significant differences in role function,emotional function and social function in intervention group when compared with nursing before (P >0.05),but the scores of rest programs of European Organization Research and Treatment of Cancer (EORTC)Quality of Life Questionnaire (QLQ)-30 decreased significantly than nursing before (P <0.01),and intervention group was significantly lower in each score of above programs than control group (P <0.05 or P <0.01).Conclusion Psychological nursing can effectively improve the psychological condition,reduce negative emotions and improve the quality of life in patients with malignant tumor chemotherapy.

ABSTRACT:Objective To explore the application value of psychological nursing in treatment of patients with malignant tumor chemotherapy.Methods A total of 105 patients with malignant tumor chemotherapy were randomly divided into intervention group (52 cases)and control group (53 cases).Control group was treated with routine nursing,on this basis intervention group was added with psychological nursing,including evaluation of psychological condition,establishment of favorable nurse-patient relationship,regular communication,optimization of patients’social sup-port network and relaxation training,etc.Results After nursing,self-rating anxiety scale (SAS) and self-rating depression scale (SDS)scores of both groups decreased significantly,which were significantly lower in intervention group than control group (P <0.01).After nursing,there were no significant differences in role function,emotional function and social function in intervention group when compared with nursing before (P >0.05),but the scores of rest programs of European Organization Research and Treatment of Cancer (EORTC)Quality of Life Questionnaire (QLQ)-30 decreased significantly than nursing before (P <0.01),and intervention group was significantly lower in each score of above programs than control group (P <0.05 or P <0.01).Conclusion Psychological nursing can effectively improve the psychological condition,reduce negative emotions and improve the quality of life in patients with malignant tumor chemotherapy.