Logistic regression analysis of drug compliance and influencing factors in elderly osteoporosis patients.

A cross-sectional study was conducted to understand the medication compliance of elderly osteoporosis patients and to further analyze the influencing factors of drug compliance. The elderly osteoporosis patients who were admitted to the First People's Hospital of Huzhou, Zhejiang from March 2015 to January 2017 were selected as the research subjects. Subsequently, the three months, six months and 12 months of follow-up were performed from the group of subjects who prescribe anti-osteoporosis drugs to determine the patient's medication status. Hereon, a cross-sectional survey was conducted to investigate the status of drug compliance in elderly patients with osteoporosis. Moreover, multivariate logistic regression analysis was used to analyze the influencing factors of medication compliance. The discontinuation rates and causes were not the same in different time periods and the cumulative withdrawal rate was high (37%) within one year. A total of 492 cases of elderly osteoporosis patients were included in this study, whereas only 45.73% of patients had good drug compliance. Elderly patients with osteoporosis have poor medication compliance and education, marital status, and medication types all affect medicine-taking compliance of patients. Therefore, it is suggested that health education should be carried out, and psychological care of patients should also be strengthened. Meanwhile, the follow-up system for drug compliance should be established to improve medication compliance as well world wide.

[Regulation of aquaporin 3 protein expression in amnion epithelial cells through cAMP-PKA signal pathway].

OBJECTIVE: To investigate the expression of aquaporins-3 (AQP3) in amniotic epithelial cells regulated by cyclic adenosine monophosphate-protein kinase A (cAMP-PKA) signal pathway and to explore the mechanisms of its expression. METHODS: The amniotic epithelial cells were collected from 30 patients who underwent elective caesarean sections at term with normal amniotic fluid volume and primarily cultured. The cultured cells were treated with (1)forskolin groups: different concentration (0, 2.5, 5, 50 or 100 µmol/L) of forskolin treated cells for 2 hours, and the optimal concentration of forskolin treated cells with different time (0, 1, 2, 10 or 20 hours); (2)SP-cAMP groups: different concentration (0, 2.5, 5, 50 or 100 µmol/L) of SP-cAMP treated cells for 2 hours, and the optimal concentration of SP-cAMP treated cells with different time (0, 1, 2, 10 or 20 hours); (3)H-89 groups: different concentration (0, 5, 10, 50 or 100 µmol/L) of H-89 treated cells for 2 hours, and the optimal concentration of H-89 treated cells with different time (0, 1, 2, 10 or 20 hours ). The level of intracellular cAMP and activity of PKA were detected by using ELISA, and immunohistochemistry was used to detect the localization of AQP3, the protein expression of total cAMP-response element binding protein (CREB) and phospho-CREB (p-CREB) and AQP3 were assessed by western blot analysis. Cell proliferation was assessed by cell counting kit-8 (CCK-8) assay. RESULTS: (1) The brown staining of AQP3 was detected in both cell membrane and cytoplasm in each group. (2) There was no significant change of the cell proliferation rate among groups with different concentration of forskolin, SP-cAMP and H-89 treatment (P > 0.05). (3) After different concentration of forskolin treated 2 hours, the expression of total CREB had no significant difference among them(P > 0.05). While the expression of cAMP level, PKA activity, p-CREB and AQP3 protein were significantly changed, which were higher in 2.5 µmol/L, 5 µmol/L, 50 µmol/L forskolin group when compared with 0 µmol/L (P < 0.05). Their expressions in 5 µmol/L forskolin group were higher than that in 2.5 µmol/L and 50 µmol/L (P < 0.05). The optimal forskolin concentration was 5 µmol/L. (4) After different concentration of SP-cAMP treated 2 hours, the expression of total CREB and cAMP level had no significant difference among them (P > 0.05), while the expression of PKA activity, p-CREB and AQP3 protein were significantly changed, which were higher in 5 µmol/L, 50 µmol/L SP-cAMP group when compared with 0 µmol/L (P < 0.05). Their expressions in 50 µmol/L SP-cAMP group were higher than that in 5 µmol/L (P < 0.05). The optimal SP-cAMP concentration was 50 µmol/L. (5) After different concentration of H-89 treated 2 hours, the expression of total CREB and cAMP level had no significant difference among them (P > 0.05), while the expression of PKA activity, p-CREB and AQP3 protein were significantly changed, which were lower in 10 µmol/L, 50 µmol/L and 100 µmol/L H-89 group when compared with 0 µmol/L (P < 0.05). Their expressions in 10 µmol/L H-89 group were lower than that in 50 µmol/L, 100 µmol/L (P < 0.05). The optimal H-89 concentration was 10 µmol/L. (6) p-CREB and AQP3 protein expression were significantly lower in 5 µmol/L forskolin combined 10 µmol/L H-89 incubating 2 hours group when compared with 5 µmol/L forskolin, but higher than that in 10 µmol/L H-89 treated group (P < 0.05). Total CREB was no significant difference among the three groups (P > 0.05). CONCLUSION: cAMP-PKA signal transduction pathway may regulate AQP3 protein expression in human amniotic epithelial cells.

[Effects of compound salvia miltiorrhiza injection on aquaporin 3 expression in human amniotic epithelial cells].

OBJECTIVE: To study the effects of Compound Salvia miltiorrhiza Injection (CSI) on aquaporin 3 (AQP3) expression in human amniotic epithelial cells (hAECs), and to explore its mechanisms for treating oligohydramnios. METHODS: The hAECs selected from 8 human term pregnancies with oligohydramnios and no other complications (as the test group)and 8 human term pregnancies with normal amniotic fluid volume (as the control group) were primarily cultured. The mRNA and protein expressions of AQP3 in hAECs were detected using reverse transcription-polymerase chain reaction and Western blot with various concentrations of CSI (0.000, 0.001, 0.010, 0.020, 0.060, and 0.100 mg/mL, respectively) at different time points (0, 6, 12,24, and 48 h, respectively). RESULTS: (1) Compared with the control group, the AQP3 expression was down-regulated in the test group (P < 0.05). (2) The AQP3 expression in the two groups reached the peak when the concentration of CSI was 0.010 mg/mL, showing statistical difference when compared with other concentrations (P < 0.05). (3) The AQP3 expression reached the peak when 0.010 mg/mL CSI acted for 12 h, showing statistical difference when compared with other concentrations (P < 0.05). (4) The AQP3 expression was up-regulated in the two groups when 0.010 mg/mL CSI acted for 12 h. But the up-regulated AQP3 expression was more obvious in the test group than in the control group (P < 0.05). CONCLUSIONS: CSI could regulate the AQP3 expression in hAECs. CSI showed more obvious effects on the AQP3 expression in hAECs of oligohydramnios human term pregnancies.

Immunohistochemical expression of Annexin A2 and S100A proteins in patients with bulky stage IB-IIA cervical cancer treated with neoadjuvant chemotherapy.

OBJECTIVE: Abnormal expression of Annexin A2 and S100A proteins has been reported to induce sensitivity/resistance to chemotherapy in a variety of cancers. The aim of this study was to evaluate the significance of Annexin A2 and S100A protein expression to predict response to neoadjuvant chemotherapy and prognostic significance of these protein expressions in bulky stage IB-IIA cervical cancer patients. METHODS: Paired tumor samples (pre- and post-chemotherapy) were obtained from 68 patients who were treated with cisplatin-based neoadjuvant chemotherapy and radical hysterectomy at our hospital from 2006 to 2011. The expression of Annexin A2 and S100A proteins was analyzed by immunohistochemistry. RESULTS: Thirty-six patients were identified as chemotherapy-response and 32 were non-response. (a). Protein expression in tumor cells: (1). Exposure of tumor cells to chemotherapy results in a change of Annexin A2 and S100A expression (P<0.05). (2). Annexin A2, S100A8 and S100A9 protein expression correlates with tumor response to chemotherapy (P<0.05). (b). Protein expression in stromal cells: (1). Expression of Annexin A2, S100A8 and S100A9 was increased, but S100A2 and S100A4 was decreased after exposure to chemotherapy (P<0.05). (2). Only S100A4 expression was associated with response to chemotherapy (P<0.05). Multivariate analysis revealed that tumor size (P=0.022), differentiation (P=0.000), Annexin A2 expression in stromal cells (P=0.009), and S100A8 expression in tumor cells (P=0.008) were independent prognostic factors for progression-free survival of cervical cancer patients. CONCLUSIONS: Expression of some of the measured proteins in tumor and stromal cells correlates with chemotherapy exposure, response to therapy, and progression-free survival.

ADH7, miR-3065 and LINC01133 are associated with cervical cancer progression in different age groups.

The aim of the present study was to identify potential therapeutic targets that serve crucial roles in the progression of cervical cancer. Clinical data, RNA sequencing (RNAseq)-counts and micro (mi)RNA data regarding cervical squamous cell carcinoma were retrieved from The Cancer Genome Atlas, and analyses were performed using the University of California Santa Cruz database. RNAseq and miRNA data were stratified into 3 groups (according to the patients' age), and genes were re-annotated and preprocessed prior to Mfuzz time clustering analysis. Subsequently, enrichment analyses were performed in order to identify differentially expressed mRNAs (DEmRNAs) and a protein-protein interaction analysis network was constructed. miRNA-gene, miRNA-lncRNA, and long non-coding (lnc)RNA-mRNA pairs were collected and the lncRNA-miRNA-mRNA competing endogenous (ce)RNA network was established. Further enrichment analyses were performed in order to identify crucial mRNAs in the ceRNA network. Finally, survival and drug association analyses were implemented. A total of 269 DEmRNAs [including alcohol dehydrogenase 7 (ADH7), vestigial-like family member 3 (VGLL3) and cytochrome P450, family 26, subfamily B, polypeptide 1 (CYP26B1)], 274 DElncRNAs (including LINC01133) and 16 DEmiRNAs (including miR-3065 and miR-330) were identified. There were 102 lncRNAs, 15 miRNAs, 15 mRNAs and 522 interaction pairs in the ceRNA network. In particular, ADH7 was regulated by miR-3065, and miR-3065 interacted with LINC01133 in the ceRNA network. Furthermore, ADH7 and CYP26B1 were enriched in the retinoic acid metabolic process and the retinol metabolism pathway. ADH7 and VGLL3 were significantly associated with the cervical cancer survival rate. ADH7, VGLL3, CYP26B1, miR-3065, miR-330, miR-499a and LINC01133 play pivotal roles in the progression of cervical cancer in different age groups.

Tanshinone IIA increases aquaporins expression in human amniotic epithelial WISH cells by stimulating GSK-3ß phosphorylation.

BACKGROUND: A recent study found that Salvia miltiorrhiza extract substantially increased amniotic fluid index in women with oligohydramnios. Abnormal aquaporin (AQP) expression is associated with oligohydramnios. This study aims to investigate the effect of tanshinone IIA, the major lipophilic component in the root of Salvia miltiorrhiza, on the function and expression of AQPs in human amniotic epithelial WISH cells and the molecular mechanism underlying the effect. METHODS: WISH cells were treated with tanshinone IIA. Cell proliferation and swelling were determined by cell counting kit-8 assay and measurement of cell size, respectively. A human phospho-MAPK antibody array was used to screen tanshinone IIA-associated signaling molecules. The protein expression of AQPs, glycogen synthase kinase 3ß (GSK-3ß), and phosphorylated GSK-3ß (p-GSK-3ß) was analyzed by Western blot. RESULT: Tanshinone IIA significantly inhibited WISH cell proliferation at ≥50µM (P<0.05) and the IC50 was 35µM. Cell size was significantly increased by the treatment with 35µM tanshinone IIA for 24h (P<0.05). WISH cells expressed AQP1, AQP3, AQP8, and AQP9 predominantly in cytoplasm. The dose of tanshinone IIA to maximally induce AQP1 and AQP3 expression was 35µM; for AQP8 and AQP9, the dose of maximal stimulation was 5µM (All P<0.05). Tanshinone IIA (35µM)-mediated up-regulation of AQP8, AQP9, AQP1, and AQP3 peaked at 6, 12, 24, and 24hours of treatment, respectively. Tanshinone IIA increased p-GSK-3ß level significantly (P<0.05). Blockade of GSK-3ß expression by siRNA and inhibition of GSK-3ß activity by lithium chloride significantly reduced tanshinone IIA-induced AQP1 and AQP3 expression (All P<0.05). CONCLUSION: Tanshinone IIA increases AQPs expression in WISH cells by stimulating p-GSK-3ß. AQP8 responds to tanshinone IIA the most rapidly compared with AQP9, AQP1, and AQP3 in WISH cells.

Expression of AQP3 protein in hAECs is regulated by Camp-PKA-CREB signalling pathway.

Previous studies by others and our group have demonstrated the expression of AQP3 protein in human chorioamniotic membranes. Here, we investigated whether cyclic adenosine monophosphate (cAMP) up-regulation of aquaporin 3 (AQP3) protein expression in human amniotic epithelial cells (hAECs) is mediated by protein kinase A (PKA) dependent or independent pathway. Cells were treated with various concentrations of Forskolin, SP-cAMP, H-89 at various time intervals or optimal concentration of Forskolin in combination with H-89 in the blocking experiments. Forskolin significantly increased cAMP levels and the expression of PKA, p-CREB and AQP3. SP-cAMP had similar effects. H-89 inhibited PKA, p-CREB and AQP3 protein expression, and attenuated the stimulatory effect of Forskolin. These results show that the AQP3 protein expression in hAECs was regulated by cAMP-PKA-CREB signal pathway. A relatively short biological half life of AQP3 renders its rapid responsiveness to regulation.

The expression and regulation of aquaporins in placenta and fetal membranes.

Previous studies by our group as well as other researchers have found expression of Aquaporins (AQPs) 1, 3, 8, and 9 in human chorioamniotic membranes and placenta. Our previous study found that the alteration of the expression of AQPs 1, 3, 8, and 9 in placenta and fetal membranes was an adaptive response to maintain amniotic fluid homeostasis in case of abnormal amniotic fluid volume, which is likely to affect the intramembranous absorption and transport of water and solute from mother to fetus. However, the actual regulation mechanisms of intramembranous absorption and placental water flow are not yet clear, making it difficult to treat abnormal amniotic fluid volume effectively. Several studies found that many factors, including hormone levels, osmotic pressure, temperature, and oxygen concentration, regulate expression of AQPs in placenta, fetal membranes, and other mammalian organs through several signal transduction pathways, such as the cAMP, the MAPK, the PI3K/AKT, and the PKC pathways. These factors could provide potential therapeutic targets for the treatment of abnormal amniotic fluid volume.

Expression and localization of aquaporins 8 and 9 in term placenta with oligohydramnios.

To test the expression and localization of aquaporins 8 (AQP8) and 9 (AQP9) in human term fetal membranes and placenta in both oligohydramnios and normal amniotic fluid volume (AFV) groups and to explore the association between aquaporin expression and oligohydramnios. Real-time polymerase chain reaction and immunohistochemistry were used to determine AQP8 and AQP9 expression levels and localization in amnion, chorion, and placenta, respectively. In addition, compared with the normal AFV group, the expression levels of both AQP8 and AQP9 in amnion in oligohydramnios group were significantly decreased, while their expressions in placenta were significantly increased. The expression level of AQP9 was also significantly decreased in chorion, while that of AQP8 was unchanged. Both AQP8 and AQP9 may play an important role in water flow both in intramembranous absorption and in placental water transfer. Our study offers the potential therapeutic approach for oligohydramnios.