Molecular mechanism of Chang Shen Hua volatile oil modulating brain cAMP-PKA-CREB pathway to improve depression-like behavior in rats.

BACKGROUND: Depression is a common and complex mental illness that manifests as persistent episodes of sadness, loss of interest, and decreased energy, which might lead to self-harm and suicide in severe cases. Reportedly, depression affects 3.8 % of the world's population and has been listed as one of the major global public health concerns. In recent years, aromatherapy has been widely used as an alternative and complementary therapy in the prevention and treatment of depression; people can relieve anxiety and depression by sniffing plant aromatic essential oils. Acorus tatarinowii and Panax ginseng essential oils in Chang Shen Hua volatile oil (CSHVO) are derived from Acorus tatarinowii and Panax ginseng, respectively, the main medicines in the famous Chinese medicine prescription Kai Xin San (KXS), Then, these oils are combined with the essential oil of Albizia julibrissin flower to form a new Chinese medicine inhalation preparation, CSHVO. KXS has been widely used in the treatment of depression; however, whether CSHVO can ameliorate depression-like behavior, its pharmacological effects, and the underlying mechanisms of action are yet to be elucidated. STUDY DESIGN AND METHODS: A rat model of chronic and unpredictable mild stimulation (CUMS) combined with orphan rearing was treated with CSHVO for 4 weeks. Using behavioral tests (sucrose preference, force swimming, tail suspension, and open field), the depression-like degree was evaluated. Concurrently, brain homogenate and serum biochemistry were analyzed to assess the changes in the neurotransmitters and inflammatory and neurotrophic factors. Furthermore, tissue samples were collected for histological and protein analyses. In addition, network pharmacology and molecular docking analyses of the major active compounds, potential therapeutic targets, and intervention pathways predicted a role of CSHVO in depression relief. Subsequently, these predictions were confirmed by in vitro experiments using a corticosterone (CORT)-induced PC12 cell damage model. RESULTS: CSHVO inhalation can effectively improve the weight and depression-like behavior of depressed rats and regulate the expression of inflammatory factors and neurotransmitters. Hematoxylin-eosin, Nissl, and immunofluorescence staining indicated that compared to the model group, the pathological damage to the brain tissues of rats in the CSHVO groups was improved. The network pharmacological analysis revealed that 144 CSHVO active compounds mediate 71 targets relevant to depression treatment, most of which are rich in the cAMP signaling and inflammatory cytokine pathways. Protein-protein interaction analysis showed that TNF, IL6, and AKT are the core anti-depressive targets of CSHVO. Molecular docking analysis showed an adequate binding between the active ingredients and the key targets. In vitro experiments showed that compared to the model group, the survival rate of PC12 cells induced by CSHVO intervention was increased, the apoptosis rate was decreased, and the expression of inflammatory cytokines in the cell supernatant was improved. Western blot analysis and immunofluorescence staining confirmed that CSHVO regulates PC12 cells in the CORT model through the cAMP-PKA-CREB signaling pathway, and pretreatment with PKA blocker H89 eliminates the protective effect of CSHVO on CORT-induced PC12 cells. CONCLUSIONS: CSHVO improves CORT-induced injury in the PC12 cell model and CUMS combined with orphan rearing-induced depression model in rats. The antidepressant mechanism of CSHVO is associated with the modulation of the cAMP-PKA-CREB signaling pathway.

Characteristics and survival of patients with gynecological cancers who refuse radiotherapy: a retrospective cohort study.

BACKGROUND: Radiotherapy improves survival for many cancer patients. However, some patients still refuse radiotherapy despite the recommendations of their physicians. We aimed to investigate the impact of refusing recommended radiotherapy on overall survival in patients with gynecological cancers (GC) and attempted to describe what characteristics are associated with the refusal of radiotherapy. METHODS: Data were extracted from the Surveillance, Epidemiology and End Result (SEER) database for patients who were diagnosed with GC and recommended for radiotherapy between 1988 and 2016. Kaplan-Meier and multivariate Cox regression analyses were utilized to analyze the impact of refusal of radiotherapy on overall survival. Univariate and multivariate logistic regression analyses were used to identify characteristics associated with refusal of radiotherapy. RESULTS: In total, 1,226 of 208,093 patients (0.6%) refused radiotherapy. Multivariate Cox regression analysis showed that refusal of radiotherapy was associated with poorer overall survival in GC patients with stage I/II [hazard ratio (HR) = 1.64; 95% confidence interval (CI), 1.50-1.79], but may not affect overall survival in patients with stage III/IV (HR = 1.03; 95%CI, 0.84-1.25). Multivariate logistic regression analysis demonstrated that factors such as older age (40-65 years, > 65 years), unmarried status (divorced, single, widowed), higher foreign-born rate (1.87-2.82%, 1.51-2.19), refusal of surgery (recommended but not performed), and higher grade (poorly differentiated, undifferentiated/anaplastic) may increase the likelihood of refusing radiotherapy (all P < 0.05). Factors that may reduce the likelihood of refusing radiotherapy include higher income (> 42,810$), lower grade (well-differentiated), primary site of ovarian cancer, and no/unknown chemotherapy (all P < 0.05). CONCLUSION: Refusal of radiotherapy is related to worse overall survival in GC patients with stage I/II, and many characteristics may affect a patient's choice of refusal of radiotherapy.

The effect of 2,2',4,4'-Tetrabromodiphenyl ether (BDE-47) on locomotor behaviour and muscle physiology of the sea cucumber Apostichopus japonicus.

The 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) is the predominant congener of polybrominated diphenyl ethers, and it is also a persistent organic pollutant that with a higher detection rate in samples from environment and animals. To date, there have been few studies of the effects of BDE-47 on locomotion in sea cucumbers. In this study, we investigated the influence of different concentrations of BDE-47 (low: 0.1 µg/L; moderate: 1.0 µg/L; high: 10.0 µg/L) on locomotion of Apostichopus japonicus and evaluated changes in their muscle physiology using ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry. The behavioural studies showed that the average and maximum velocity of movement decreased significantly in both the moderate and high BDE-47 groups after 1 day of exposure. In addition, levels of 55 metabolites were identified and characterized in the longitudinal muscle of A. japonicus exposed to BDE-47. The alteration of taurine and norepinephrine levels indicated that BDE-47 had drastic physiological effects on the longitudinal muscle of A. japonicus.

MicroRNA­193a­5p exerts a tumor suppressive role in epithelial ovarian cancer by modulating RBBP6.

Epithelial ovarian cancer (EOC), a gynecological tumor, is associated with high mortality. MicroRNAs (miRs) serve a crucial role in EOC; however, the mechanisms underlying the effect of miRNA­193a­5p in EOC are not completely understood. Therefore, the present study aimed to investigate the expression levels of miR­193a­5p in serum samples of patients with EOC and to determine the role of miR­193a­5p in EOC. Reverse transcription­quantitative PCR was used to analyze the expression levels of miR­193a­5p in serum samples of patients with EOC and EOC cell lines. The effects of miR­193a­5p and RB binding protein 6, ubiquitin ligase (RBBP6) on the biological functions of EOC were determined by conducting a series of in vitro cell function experiments. The results indicated that the expression levels of miR­193a­5p were significantly decreased in serum samples obtained from patients with EOC and EOC cell lines compared with healthy individuals and normal cells, respectively. Further investigations indicated that RBBP6 was a target gene of miR­193a­5p. The expression levels of RBBP6 were significantly increased in patients with EOC compared with healthy individuals. In addition, in vitro analysis suggested that miR­193a­5p mimic significantly decreased SKOV3 cell proliferation, migration and invasion, and promoted SKOV3 cell apoptosis compared with the control and mimic­negative control groups. In addition, RBBP6 overexpression reversed miR­193a­5p mimic­mediated effects. In conclusion, the results of the present study suggested that downregulated expression levels of miR­193a­5p may serve an inhibitory role in EOC by inhibiting cell proliferation and metastasis, and promoting apoptosis.

Retracted: The Effects of Ludartin on Cell Proliferation, Cell Migration, Cell Cycle Arrest and Apoptosis Are Associated with Upregulation of p21WAF1 in Saos-2 Osteosarcoma Cells In Vitro.

The authors repeated experiments and found that the results shown in figure 2 were not reproducible. Reference: Shuang-li Zhang, Bao-lin Li, Wei Li, Ming Lu, Lin-ying Ni, Hui-li Ma, Qing-gang Meng. The Effects of Ludartin on Cell Proliferation, Cell Migration, Cell Cycle Arrest and Apoptosis Are Associated with Upregulation of p21WAF1 in Saos-2 Osteosarcoma Cells In Vitro. Med Sci Monit 2018; 24: LBR4926-4933. 10.12659/MSM.909193.

The effects of kanamycin concentration on gene transcription levels in Escherichia coli.

In the area of microbial production of valuable chemicals, plasmids have been widely applied for overexpressing rate-limiting enzymes with high yields. However, the effect of antibiotic concentrations on the transcription of target genes in E. coli is less involved in previous reports. In this study, we constructed E. coli strains expressing the reporter gene and the kanamycin resistant gene in an operon, and analyzed the transcription levels of the reporter gene and the fluorescent intensity of the recombinant E. coli under different kanamycin concentrations. We found that the growth and gene transcription of the recombinant strain were affected obviously by the concentration of kanamycin, indicating the importance of fine-tuning of antibiotic concentrations in microbial fermentation.

AAV-mediated siRNA against TRPV1 reduces nociception in a rat model of bone cancer pain.

Objective: Bone cancer pain is characterized by moderate to severe ongoing pain that commonly requires the use of opiates. Transient receptor potential vanilloid subfamily member 1 (TRPV1), a new target of the analgesics, activated by heat, protons and capsaicin and the hot component of pepper. However, little is known of the anti-nociceptive effects of TRPV1 in cancer-induced bone pain. RNA interference (RNAi) has proven to be a powerful technique to study the function of genes by producing knock-down phenotypes. The aim of this study is to investigate the potential role of TRPV1 in rat model of bone cancer pain. Methods: Bone cancer pain animal model was created by tumor cell implantation (TCI). An AAV-mediated siRNA against TRPV1 was intrathecally delivered into the rats. Animal behaviors were measured using a set of mechanical or electronic von Frey apparatus and hot plate. mRNA and protein expression were examined by using qPCR and western blot methods. Results: Mechanical threshold and paw withdrawal latency in response to thermal stimulation were significantly elevated in rats with intrathecal administration of AAV-mediated siRNA against TRPV1. Moreover, class I histone deacetylases (HDACs), which plays a critical role in the neuro-inflammation response, and TNFα in the spinal cord were also significantly suppressed upon knockdown of TRPV1 by AAV-mediated siRNA against TRPV1 in rat spinal cord. Conclusions: Knockdown of TRPV1 effectively ameliorated mechanical allodynia and thermal hyperalgesia induced by TCI. Our data demonstrated that modulate the expression of TRPV1 in the spinal cord could be a potential therapeutic approach for bone cancer pain.

The Effects of Ludartin on Cell Proliferation, Cell Migration, Cell Cycle Arrest and Apoptosis Are Associated with Upregulation of p21WAF1 in Saos-2 Osteosarcoma Cells In Vitro.

BACKGROUND The aim of this study was investigate the effects of the sesquiterpene lactone, ludartin, on cell proliferation, cell migration, apoptosis, and the cell cycle in osteosarcoma cell lines, compared with a normal osteoblast cell line. MATERIAL AND METHODS Osteosarcoma cell lines, MG-63 Saos-2 U-2OS, T1-73 143B, and HOS, and normal hFOB 1.19 osteoblasts, were cultured and treated with increasing doses of ludartin, The MTT colorimetric assay was used to measure cell metabolic activity and viability. Apoptosis was studied by fluorescence-activated cell sorting (FACS) using 4',6-diamidino-2-phenylindole (DAPI) nuclear staining and Annexin-V/propidium iodide (PI) staining. Cell cycle was studied using flow cytometry. Cell migration and invasion were studied using wound healing and Boyden chamber assays. Protein expression was measured by Western blotting. RESULTS Ludartin inhibited cell viability, cell migration, cell proliferation, and increased cell apoptosis, in all osteosarcoma cell lines, with an IC50 dose ranging from 15-30 µM. The greatest effects were on the Saso-2 osteosarcoma cells, with an IC50 of 15 µM. However, ludartin showed minor cytotoxic effects of the normal hFOB 1.19 osteoblasts (IC50 >100 µM). Ludartin exerted its anti-proliferative effects on Saos-2 cells via induction of apoptosis and cell cycle arrest at the G2/M checkpoint, associated with reduced expression of Cdc25c (Ser216), Cdc25c, pCdc2 (Tyr15), and Cdc2 and increased expression of p21WAF1. Ludartin inhibited cell migration and invasion of the Saos-2 cells. CONCLUSIONS The dose-dependent effects of ludartin on cell proliferation, migration, apoptosis, cell cycle arrest at the G2/M checkpoint involved p21WAFI in Saos-2 osteosarcoma cells.

Effects of connective tissue growth factor on prostate cancer bone metastasis and osteoblast differentiation.

Previous studies have demonstrated that connective tissue growth factor (CTGF) is expressed at increased levels in prostate cancer bone metastasis mouse models and patients with prostate cancer which metastasizes to the bone; however, the underlying molecular mechanism(s) remain unknown. The present study investigated the function of CTGF in osteoblast differentiation and its effect on prostate cancer bone metastasis by analyzing CTGF gene expression and transcription at different levels of invasion, metastasis of prostate cancer cells, and the influence of CTGF on proliferation and xenotransplantation. A mouse model demonstrating bone metastasis was used to investigate the function(s) of CTGF in bone metastasis and osteoblast differentiation. Results demonstrated that CTGF expression was increased in association with high bone metastasis in prostate cancer cells, and its expression was significantly decreased in whole cell lysates. CTGF expression in prostate cancer cells with high levels of bone metastasis was increased 1.9-fold compared with prostate cancer cells with low levels of bone metastasis. The expression of CTGF in mesenchymal cells was markedly increased compared with epithelial cells. Results indicated that the increased expression of CTGF does not affect the proliferation of tumor cells and possesses no influence on tumor volume. Control and CTGF plasmids were transfected into RM1 cells and led to 4 and 17% bone lesions, respectively. Increased expression of CTGF significantly enlarged the tumor area in the bone metastatic position compared with the control. Positive areas of alkaline phosphatase were significantly decreased as the concentration of CTGF increased. The results of the present study demonstrated that CTGF promotes prostate carcinoma to metastasize in the bone by dysregulating osteoblast differentiation.

Relationship between RAS Association Domain Family Protein 1A Promoter Methylation and the Clinicopathological Characteristics in Patients with Ovarian Cancer: A Systematic Meta-Analysis.

BACKGROUND: To investigate the relationship between RAS association domain family protein 1A (RASSF1A) promoter methylation and the clinical features, and the survival of ovarian cancer patients. METHODS: A comprehensive literature search was conducted in the PubMed, Embase, EBSCO, and Cochrane Library databases. The overall ORs with their 95% CIs were calculated in this meta-analysis. RESULTS: Finally 17 relevant publications with 1,108 ovarian cancer samples were available for the current meta-analysis. RASSF1A promoter methylation had a significantly higher level in ovarian cancer than in low malignant potential (LMP) tumors. No significant relationship was observed between RASSF1A promoter methylation and the clinicopathological characteristics in ovarian cancer. Two studies reported that RASSF1A promoter methylation was not correlated with the survival of patients with ovarian cancer. CONCLUSIONS: Our findings suggest that the use of RASSF1A promoter methylation could distinguish ovarian cancer and LMP tumors. -RASSF1A promoter methylation may not be correlated with the clinical features and the survival of ovarian cancer patients. More studies with large sample sizes are essential in the future.

Effects of hUCB-MSCs on recovery of neurological function and TERT expression in brain tissue of rats with cerebral ischemia-reperfusion injury.

The aim of this study was to investigate and analyze the effects of human umbilical cord blood mesenchymal stem cells (hUCB-MSCs) on the recovery of neurological function and telomerase reverse transcriptase (TERT) expression in brain tissue of rats with cerebral ischemia-reperfusion injury. A total of 100 healthy adult Wistar rats were randomly divided into two groups: The control group and the observation group according to the random number table method. After the model of cerebral ischemia-reperfusion injury was established, the rats in the observation group were treated with hUCB-MSCs (10 ml/kg), while the rats in the control group were treated with saline every day. The neurological deficit score and foot fault test were evaluated at 1, 7 and 14 days after treatment, and the rats were sacrificed at 14 days to detect the expression of TERT in brain tissue. There was no significant difference in the scores of mNSS between the two groups before the model establishment (P>0.05), but there was significant differences in two groups after the operation (P<0.05). At 1 day after the operation, the mNSS score of the two groups peaked, which was decreased in the groups with the progress of treatment. The degree of decline in the observation group was significantly greater than that in the control group (P<0.05). Similarly, there was no significant difference in the number of errors between the two groups before the model establishment (P>0.05), but there was significant difference in two groups after the operation (P<0.05). At 1 day after the operation, the number of errors also peaked, which was reduced in the groups with the progress of treatment. The degree of reduction in the observation group was significantly greater than that in the control group (P<0.05). The results of H&E staining showed it had positive reaction as nucleus or cytoplasm stained brown or yellowish brown in the observation group, while it showed neuronal shrinkage, cytoplasm and nucleus yellow dye deepening in the control group as the significant positive reaction. The gray level of the TERT protein in the brain tissue of the control group was 0.458±0.052 LOD, which was significantly lower than that in the observation group with 0.983±0.056 LOD (P<0.05). In conclusion, hUCB-MSCs can effectively improve the neurological function and the expression of TERT in brain tissue of rats with cerebral ischemia-reperfusion injury, which may be helpful to reduce the ischemia-reperfusion injury of brain tissue.

Effects of pulsed electromagnetic fields on histomorphometry and osteocalcin in disuse osteoporosis rats.

OBJECTIVE: Bone histomorphometry and the concentration of the serum osteocalcin expression were observed in disuse osteoporosis (DOP) rats in order to explore the mechanism of pulsed electromagnetic fields in treating DOP. METHODS: Female SD rats, weighing 250 ∼ 280 g, were randomly divided into 4 groups: a control group and three experimental groups. The right hindlimbs of all the rats were immobilized by tibia-tail fixation, except for those in the INT group. The ALN group rats were given an alendronate sodium (1 mg kg-1 d-1) treatment, and the rats in the PEMF group received PEMF irradiation. Bone histomorphometry and the concentration of serum osteocalcin expression were detected in 2, 4, 8, and 12 weeks. RESULTS: Four weeks after modeling, as compared with the DOP group, the %Tb-Ar and Tb-N in the ALN group were increased, and the difference was significant (P< 0.05 or P< 0.01). At 8 weeks, as compared with the DOP group, the %Tb-Ar and Tb-N in the ALN group and the PEMF group both increased, and there was a significant difference (P< 0.05 or P< 0.01). At 12 weeks, as compared with the DOP group, the BGP serum concentration of the ALN group was reduced, and there was a statistical difference (P< 0.05). CONCLUSION: Pulse electromagnetic field with drug can prevent disuse osteoporosis by changing the bone microstructure. In the long run, PEMF improves the mechanical performance of biological structures better than alendronate sodium does. PEMF may influence the process of bone remodeling by promoting the level of osteocalcin.