Asperosaponin VI Protects Against Bone Loss Due to Hindlimb Unloading in Skeletally Growing Mice Through Regulating Microbial Dysbiosis Altering the 5-HT Pathway.

Osteoporosis is a complex multifactorial disease that can lead to an increased risk of fracture. However, selective and effective osteoporosis drugs are still lacking. We showed that Asperosaponin VI (AVI) has the implications to be further developed as an alternative supplement for the prevention and treatment of bone loss. AVI has been found to have beneficial effects on metabolic diseases such as bone loss, obesity, and atherosclerosis. Our study was designed to determine the effect and mechanism of action of AVI against bone loss through regulating microbial dysbiosis. A hindlimb unloading mouse model was established to determine the effect of AVI on bone microarchitecture, gut microbiota, and serum metabolites. Eighteen female C57BL/6 J mice were divided into three groups: control, hindlimb unloading with vehicle (HLU), and hindlimb unloading treated with AVI (HLU-AVI, 200 mg/kg/day). AVI was administrated orally for 4 weeks. The results demonstrated that AVI improved the bone microstructure by reversing the decrease in bone volume fraction and trabecular number, and the increase in trabecular separation and structure model index of cancellous bone in hindlimb suspension mice. The results of 16sRNA gene sequencing suggested that the therapeutic effect of AVI on bone loss may be achieved through it regulating the gut microbiota, especially certain specific microorganisms. Combined with the analysis of ELISA, immunohistochemistry, and serum metabolome results, it could be speculated that AVI played an important role in adjusting the balance of bone metabolism by influencing specific flora such as Clostridium and its metabolites to regulate the 5-hydroxytryptophan pathway. The study explored the novel mechanism of AVI against osteoporosis, and has implications for the further development of AVI as an alternative supplement for the prevention and treatment of bone loss.

Upregulation of FoxO6 in nucleus pulposus cells promotes DNA damage repair via activation of RAD51.

OBJECTIVE: DNA damage is an essential risk for intervertebral disc degeneration (IDD). Here, we attempted to uncover the effect of FoxO6 and RAD51 on the DNA damage repair of nucleus pulposus (NP) cells in IDD. PATIENTS AND METHODS: We collected the human NP tissues of different degeneration degrees and tested the collagen II, FoxO6, and RAD51 expression. Besides, the IL-1ß induced NP cell model was also used to elucidate the degenerative progress in vitro. We used Chromatin immunoprecipitation (ChIP) and luciferase reporter assay to confirm whether the FoxO6 protein could enhance the RAD51 expression by binding to its promoter. The FoxO6 gene was upregulated in NP cells by vectors transfection. Immunofluorescence staining was used to measure the RAD51 and γH2AX foci formation. Besides, the typical NP cell gene expression was analyzed by RT-PCR. Cell proliferation was determined by CCK-8, and the cell cycle distribution was determined by flow cytometry. RESULTS: Like collagen II, FoxO6 and RAD51 expression were all decreased both in the severe degenerated NP tissue and in the IL-1ß treated NP cells. Upregulation of FoxO6 gene in NP cells enhanced the RAD51 expression via activating the promoter region and inhibited the DNA damage marker γH2AX formation. FoxO6 upregulation alleviated the loss of collagen II, aggrecan, SOD1, and CAT, and suppressed the increase of collagen I/X, TNF-α, and IL-1ß expression, which was affected by IL-1ß. Besides, FoxO6 also helped the proliferation and cell cycle of NP cells with the activation of RAD51. CONCLUSIONS: Upregulation of FoxO6 promotes the DNA repair and maintains the typical phenotype of NP cells, via somehow the mediation of RAD51.

Protective effect of MAPK signaling pathway mediated by ITGB3 gene silencing on myocardial ischemia-reperfusion injury in mice and its mechanism.

OBJECTIVE: To explore the effect of integrin ß3 (ITGB3) gene silencing mediated mitogen-activated protein kinase (MAPK) signaling pathway on myocardial ischemia-reperfusion injury (MIRI) in mice. MATERIALS AND METHODS: MIRI mice model was established, and myocardial tissues of MIRI mice and sham operation group mice were extracted. Hematoxylin-Eosin (HE) staining was used to observe the pathological changes of myocardial tissue; terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) staining was used to detect the apoptosis of myocardial cells; ELISA method was used to detect the levels of interleukin (IL)-1, IL-6, and tumor necrosis factor (TNF)-α in the two groups. The infarct size was measured by TTC staining. Myocardial cells of MIRI model mice were isolated and cultured, and then grouped and transfected. The cells were transfected with the grouping of MIRI group, negative control (NC) group, MAPK signal pathway agonist Anisomycin group, MAPK signal pathway inhibitor SB203580 group, ITGB3-siRNA group, SB203580 + ITGB3-siRNA group. Real-time quantitative polymerase chain reaction (qRT-PCR) and Western blot were used to detect the mRNA and protein expressions of ITGB3, p38MAPK/p-p38MAPK, GSK-3ß/p-GSK-3ß, Cx43/p-Cx43, pro-apoptotic factor Bax and anti-apoptotic factor Bcl-2. 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide (MTT) assay was used to detect cell proliferation and flow cytometry to detect cell apoptosis. RESULTS: The expression of ITGB3 in myocardial tissue of MIRI mice was significantly higher than that of sham operated mice (p<0.05). Compared with the sham operation group, the apoptosis rate of myocardial cells in MIRI group was significantly increased, the expression levels of IL-1, IL-6 and TNF-α were significantly increased, and the myocardial infarction area was significantly increased (all p<0.05). Compared with MIRI and NC groups, ITGB3 mRNA and protein expression levels in ITGB3-siRNA group and SB203580 + ITGB3-siRNA group were significantly decreased (all p<0.05), but no significant change was found in Anisomycin group and SB203580 group (p>0.05). Furthermore, ITGB3-siRNA group and Anisomycin group had markedly decreased mRNA and protein expressions of ITGB3 and Bax, increased mRNA and protein expressions of p38MAPK/p-p38MAPK, GSK-3ß/p-GSK-3ß, Cx43/p-Cx43 and Bcl-2, as well as increased cell proliferation and decreased cell apoptosis (all p<0.05); SB203580 group indicated an opposite result with Anisomycin group; while SB203580 + ITGB3-siRNA revealed none significant statistical difference. In addition, compared with ITGB3-siRNA group, SB203580 + ITGB3-siRNA group showed significantly upregulated mRNA and protein expressions of Bax, downregulated mRNA and protein expressions of p38MAPK/p-p38MAPK, GSK-3ß/p-GSK-3ß, Cx43/p-Cx43 and Bcl-2, as well as decreased cell proliferation and increased cell apoptosis (all p<0.05). CONCLUSIONS: Silencing ITGB3 gene expression can promote the activation of MAPK signaling pathway, elevate the phosphorylation of GSK-3ß and Cx43 in the downstream, promote the proliferation of mouse myocardial cells, inhibit myocardial cell apoptosis and inflammatory reaction, and thus have protective effect on MIRI in mice.

Dexmedetomidine promotes the recovery of renal function and reduces the inflammatory level in renal ischemia-reperfusion injury rats through PI3K/Akt/HIF-1α signaling pathway.

OBJECTIVE: To evaluate the protective effect of dexmedetomidine (Dex) against renal ischemia-reperfusion injury (RIRI) in rats through the phosphatidylinositol 3-hydroxy kinase (PI3K)/protein kinase B (Akt)/hypoxia-inducible factor-1α (HIF-1α) signaling pathway. MATERIALS AND METHODS: (1) A Sprague- Dawley rat model of RIRI was established. Thirty rats were divided into Sham group, injury (RIRI) group, and Dex treatment (RIRI + Dex) group. Serum was collected to detect renal function-related indexes, and the levels of serum inflammatory factors were examined via enzyme-linked immunosorbent assay (ELISA). (2) The kidney tissues were separated, and the degree of tissue damage was determined using immunohistochemical staining. (3) Ribonucleic acids (RNAs) were extracted from tissues, and the mRNA levels of inflammatory factors were measured using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). (4) The protein expressions of Akt, phosphorylated (p)-Akt, PI3K, p-PI3K, and HIF-1α were detected via Western blotting. RESULTS: Compared with those in RIRI group, the levels of blood urea nitrogen and creatinine declined (p<0.05), the synthesized mRNAs of inflammatory factors in the kidney tissues were reduced (p<0.05), the secreted serum inflammatory factors was also reduced (p<0.05), and the phosphorylation levels of Akt and PI3K and the HIF-1α level rose (p<0.05) in RIRI + Dex group. CONCLUSIONS: Dex promotes the recovery of renal function and reduces the inflammatory level in RIRI rats through the PI3K/Akt/HIF-1α signaling pathway.

Mechanism of miR-122-5p regulating the activation of PI3K-Akt-mTOR signaling pathway on the cell proliferation and apoptosis of osteosarcoma cells through targeting TP53 gene.

OBJECTIVE: To explore the regulatory mechanism of microRNA-122-5p (miR-122-5) targeting tumor protein p53 (TP53) gene to mediate PI3K-Akt-mTOR signaling pathway on the proliferation and apoptosis of osteosarcoma (OS) cells. PATIENTS AND METHODS: With the collection of osteosarcoma and normal adjacent tissues, the mRNA of miR-122-5p, TP53, PTEN, PI3K, Akt, mTOR, Bim, Bax, and Bcl-2 was detected by Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR), followed by the detection of the protein expression by Western blot. The target relationship between miR-122-5p and TP53 gene was verified. The third generation osteosarcoma cells were divided into Blank group, miR-122-5p mimic negative control (NC) group, miR-122-5p mimic group, miR-122-5p inhibitor NC group, miR-122-5p inhibitor group, rapamycin group and miR-122-5p inhibitor + rapamycin group. Furthermore, 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry were used to detect the proliferation ability, cell cycle distribution and apoptosis of each group after transfection. RESULTS: The expression level of miR-122-5p in osteosarcoma was lower than that in normal tissues (p < 0.05), TP53, PTEN, Bim and Bax expression levels were decreased (all p < 0.05), while the expression levels of PI3K, p-PI3K, Akt, p-Akt, mTOR, p-mTOR and Bcl-2 were highly upregulated (all p < 0.05). TP53 had the lowest expression in osteosarcoma cell line U-2OS (p < 0.05), which was selected for subsequent cell experiments. TP53 was the target gene of miR-122-5p. Compared with Blank group, miR-122-5p mimic group had increased expression of miR-122-5p (all p < 0.05); besides, there were significantly increased expression of TP53, PTEN, Bim, and Bax in miR-122-5p mimic group and rapamycin group, while remarkably decreased expression of PI3K, p-PI3K, Akt, p-Akt, mTOR, p-mTOR, and Bcl-2 (all p < 0.05), accompanied by increased proportion of cells in G0/G1 phase, decreased cell proportion in S phase, increased cell apoptosis and inhibited cell proliferation (all p < 0.05). The opposite trends were found in miR-122-5p inhibitor group relative to miR-122-5p mimic group and rapamycin group (all p < 0.05). Meanwhile, no significant difference was found in miR-122-5p inhibitor+rapamycin group when compared with that in Blank group (all p > 0.05) except for significantly decreased miR-122-5p expression (p < 0.05). CONCLUSIONS: Upregulation of miR-122-5p may inhibit the proliferation and promote the apoptosis of osteosarcoma cells by inhibiting the activation of PI3K-Akt-mTOR signaling pathway, which may be related to the targeted up-regulation of TP53 expression.

LncRNA NNT-AS1 regulates the progression of lung cancer through the NNT-AS1/miR-3666/E2F2 axis.

OBJECTIVE: Lung cancer is the main burden on human health, with high mortality and poor prognosis. The involvement of long non-coding RNAs (lncRNAs) in the development of cancer has attracted wide attention. This study aimed to investigate the role and novel mechanisms of lncRNA nicotinamide nucleotide transhydrogenase antisense RNA 1 (NNT-AS1) in the progression of lung cancer. MATERIALS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was performed to detect the expression of NNT-AS1, microRNA-3666 (miR-3666), and E2F transcription factor 2 (E2F2). 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay was used to analyze cell proliferation. Flow cytometry was carried out to investigate cell apoptosis. Transwell assay was conducted to observe cell invasion. The interaction between miR-3666 and NNT-AS1 or E2F2 was predicted by bioinformatics tool starBase v2.0 and verified by Dual-Luciferase reporter assay. The protein level of E2F2 was quantified by Western blot. RESULTS: NNT-AS1 and E2F2 were upregulated, but miR-3666 was downregulated in lung cancer tissues and cells. NNT-AS1 knockdown attenuated proliferation and invasion but enhanced apoptosis of lung cancer cells, while miR-3666 inhibition reversed these effects. It was confirmed that miR-3666 was a target of NNT-AS1 and it directly interacted with E2F2. The inhibitory proliferation and invasion, and acceleratory apoptosis of lung cancer cells, caused by miR-3666 enrichment, were overturned by E2F2 overexpression. Furthermore, E2F2 was regulated by NNT-AS1 through miR-3666. CONCLUSIONS: NNT-AS1 participated in the progression of lung cancer through NNT-AS1/miR-3666/E2F2 regulatory axis at least in part. Our study supplied a promising strategy for the treatment of lung cancer.

Reduced expression of miR-3653 in glioma and its correlations with clinical progression and patient survival.

OBJECTIVE: Growing evidence in recent years have demonstrated that the dysregulation of microRNAs (miRNAs) strongly affected the biological development and progression of human tumors, including glioma. There have been few studies on the clinical significance of miRNAs in glioma. The aim of our study is to explore the expression pattern and the value of miR-3653 in the prognosis of glioma patients. PATIENTS AND METHODS: qRT-PCR assays were performed to the miR-3653 expression level in 168 cases of glioma tissues and matched normal tissues. The correlations of miR-3653 expression level with the clinicopathological factors in glioma patients were analyzed. The associations between miR-3653 expression and survival of glioma patients were investigated by the Kaplan-Meier analysis and the log-rank test. The prognostic value of miR-3653 was estimated via univariate and multivariate analysis. RESULTS: We presented that miR-3653 level in glioma tissues is notably reduced compared to matched non-cancerous brain tissues (p<0.01). Clinical research revealed that the lower miR-3653 expression was associated with larger tumor size and lymph node metastasis, lower KPS (p=0.028), and advanced WHO grade (p=0.019). Moreover, the clinical data further suggested that the low expression of miR-3653 predicted a worse 5-year overall survival in glioma patients. Finally, the multivariate analysis confirmed that low miR-3653 expression (HR=2.682, 95% CI: 1.148-4.281, p=0.021) was a significant independent predictor of poor survival in glioma. CONCLUSIONS: Our findings suggested that miR-3653 could serve as a valuable prognostic indicator in glioma. Further researches are required to explore the potential function and mechanism of miR-3653 in glioma.

MicroRNA-889 promotes cell proliferation in colorectal cancer by targeting DAB2IP.

OBJECTIVE: Colorectal cancer (CRC) remains one of the most frequent lethal malignant tumors worldwide. The correlation between miR-889 expression and CRC progression has not been well identified in the recent literature. Here, we aim to detect the role and mechanism of miR-889 in CRC. PATIENTS AND METHODS: First, miRNA RT-PCR (Real Time-Polymerase Chain Reaction) was performed to determine miR-889 expression in CRC tissues and cells. The proliferative capacity of cells transfected with miR-889 mimics, miR-889 inhibitor or NC was measured by CCK-8 (cell counting kit-8), colony formation and EdU (5-Ethynyl-2'-deoxyuridine) assays. The online bioinformatics sites were chosen to predict possible downstream regulatory genes of miR-899. The dual-luciferase report assay was conducted to verify the relation between DAB2IP (DAB2 interacting protein) and miR-899. The expression changes of DAB2IP were assessed by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot. RESULTS: MiR-889 was upregulated in CRC tissues and CRC cells, and upregulated miR-889 was confirmed to promote cell growth in vitro. Dual-luciferase reporter, qRT-PCR, and Western blot assays suggested that DAB2IP might be regulated by miR-889. The effects of miR-889 on proliferation could be abolished by DAB2IP through confirmatory experiments. By directly targeting DAB2IP, miR-889 served as a vital part in accelerating CRC cell proliferation. CONCLUSIONS: Our current study substantiated that miR-889 might participate in controlling CRC proliferation by regulating DAB2IP, which provides potential and prospective therapeutic target for CRC.

Correlation between IL-7 genomic protein methylation level and acute myeloid leukemia.

OBJECTIVE: To detect Interleukin-7 (IL-7) gene methylation status and transcription level in leukemia cells of peripheral blood of patients with Acute Myelocytic Leukemia (AML) and in the cell lines (HL-60, HL-60/ADM, SKM-1) of AML and myelodysplastic syndrome (MDS), and explore its relationship with the pathogenesis of AML. PATIENTS AND METHODS: A total of 55 AML patients (AML group) and 30 healthy adults (Healthy group) from June 2015 to June 2018 were enrolled in this study. The genomic DNA of leukemia cells in peripheral blood was extracted. The methylation-specific PCR (MSP) method was used to detect the methylation rate of the IL-7 gene in peripheral blood of AML group and Health group. Meanwhile, the methylation level of the IL-7 gene leukemia cell lines HL-60/ADM, HL-60, and MV4-11 and SKM-1 were detected in vitro. At the same time, the expression level of IL-7 in peripheral blood was detected by Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) kit. RESULTS: The methylation rate of IL-7 gene in peripheral blood of the AML group and Healthy group was 72.7% (40/55) vs. 3.3% (1/30) (p<0.01); IL-7 gene methylation occurred in HL-60/ADM, HL-60, MV4-11 and SKM-1 cell lines. IL-7 gene methylation appears in peripheral blood leukemia cells and AML and MDS cell lines of AML patients. CONCLUSIONS: The expression of IL-7 in peripheral blood of patients with AML is significantly decreased, suggesting that this phenomenon is related to the pathogenesis of AML.

Development and validation of an individualized nomogram to identify occult peritoneal metastasis in patients with advanced gastric cancer.

BACKGROUND: Occult peritoneal metastasis (PM) in advanced gastric cancer (AGC) patients is highly possible to be missed on computed tomography (CT) images. Patients with occult PMs are subject to late detection or even improper surgical treatment. We therefore aimed to develop a radiomic nomogram to preoperatively identify occult PMs in AGC patients. PATIENTS AND METHODS: A total of 554 AGC patients from 4 centers were divided into 1 training, 1 internal validation, and 2 external validation cohorts. All patients' PM status was firstly diagnosed as negative by CT, but later confirmed by laparoscopy (PM-positive n = 122, PM-negative n = 432). Radiomic signatures reflecting phenotypes of the primary tumor (RS1) and peritoneum region (RS2) were built as predictors of PM from 266 quantitative image features. Individualized nomograms of PM status incorporating RS1, RS2, or clinical factors were developed and evaluated regarding prediction ability. RESULTS: RS1, RS2, and Lauren type were significant predictors of occult PM (all P < 0.05). A nomogram of these three factors demonstrated better diagnostic accuracy than the model with RS1, RS2, or clinical factors alone (all net reclassification improvement P < 0.05). The area under curve yielded was 0.958 [95% confidence interval (CI) 0.923-0.993], 0.941 (95% CI 0.904-0.977), 0.928 (95% CI 0.886-0.971), and 0.920 (95% CI 0.862-0.978) for the training, internal, and two external validation cohorts, respectively. Stratification analysis showed that this nomogram had potential generalization ability. CONCLUSION: CT phenotypes of both primary tumor and nearby peritoneum are significantly associated with occult PM status. A nomogram of these CT phenotypes and Lauren type has an excellent prediction ability of occult PM, and may have significant clinical implications on early detection of occult PM for AGC.

MiR-103/107 induces tumorigenicity in bladder cancer cell by suppressing PTEN.

OBJECTIVE: MiR-103/107 has been shown to be implicated in the pathogenesis of various malignant diseases. The present study was designed to analyze the expression, function and mechanism of miR-103/107 in the bladder cancer tumorigenesis. PATIENTS AND METHODS: Bladder cancer tissues and the paired normal tissues were collected during the surgical treatment of radical cystectomy, and the expression of miR-103/107 was measured by quantitative Reverse Transcriptional Polymerase Chain Reaction (RT-PCR). After modulation of miR-103/107 level in bladder cancer cells using antagomiR or mimics, several experimental approaches such as MTT assay, flow cytometry analysis and Western blot have been applied to determine cell viability, cell cycle and protein expression, respectively. Luciferase reporter assay was performed to determine the target of miR-103/107. RESULTS: miR-103/107 expression is upregulated in the tumor site of bladder cancer specimens, and it is positively associated with tumor stages. Inhibition of miR-103/107 by its antagomiR decreased the cell growth potential and induced cell cycle arrest. Moreover, inhibition of miR-103/107 also suppressed the PI3K/AKT signaling. Further analysis revealed that miR-103/107 directly targets the 3' untranslated region (UTR) of PTEN mRNA to promote PI3K/AKT signaling, which was corroborated by the negative correlation between miR-103/107 and PTEN in tumor specimens. CONCLUSIONS: The oncogenic role of miR-103/107 in bladder cancer is revealed for the first time. MiR-103/107 regulates cell proliferation and PI3K/AKT signaling partially through PTEN dependent mechanism. Thus, inhibiting miR-103/107 may be a therapeutic approach for bladder cancer treatment.

Overexpression of miR-638 attenuated the effects of hypoxia/reoxygenation treatment on cell viability, cell apoptosis and autophagy by targeting ATG5 in the human cardiomyocytes.

OBJECTIVE: Myocardial ischemia/reperfusion (I/R) injury largely contributed to the damage of myocardial tissues in patients with coronary disease, which may subsequently lead to heart failure. MicroRNAs (miRNAs) are considered to be involved in the process of myocardial I/R injury. The present study aimed to investigate the in vitro functional role of miR-638 in the myocardial I/R injury in the human cardiomyocytes (HCMs). PATIENTS AND METHODS: MTT assay and flow cytometry assay were performed to determine cell viability and apoptosis of HCMs. Real Time-quantitative Polymerase Chain Reaction was used to determine miRNA and mRNA expression levels. The protein levels were determined by Western blot assay. RESULTS: Hypoxia/reoxygenation (H/R) treatment suppressed cell viability, increased cell apoptotic rate and suppressed miR-638 expression in the HCMs. The downregulation of miR-638 suppressed cell viability and induced cell apoptosis in the HCMs. The overexpression of miR-638 attenuated the effects of H/R treatment on the cell viability and cell apoptosis in the HCMs. In addition, miR-638 suppressed the expression of autophagy-related 5 (ATG5) by targeting the 3'untranslated region of ATG5. Enforced expression of ATG5 reversed the effects of miR-638 overexpression on cell viability and cell apoptosis in H/R-treated HCMs. More importantly, H/R treatment promoted autophagy in the HCMs, and this effect was significantly reversed by miR-638 mimic transfection. CONCLUSIONS: Our results suggested that the overexpression of miR-638 attenuated the effects of H/R treatment on cell viability, cell apoptosis and autophagy, at least partly by regulating the ATG5 expression in the HCMs.

Effects of forkhead Box protein A1 on cell proliferation regulating and EMT of cervical carcinoma.

OBJECTIVE: Cervical cancer is a common tumor in gynecological malignancies. However, the patients are often in an advanced stage when diagnosed. It was found that forkhead box protein A1 (FOXA1) is abnormally expressed in various tumors, such as breast cancer, ovarian cancer, and is closely related to tumorigenesis. This study aimed to investigate the expression and the related roles of FOXA1 in cervical cancer. PATIENTS AND METHODS: Real Time-PCR (RT-PCR) and Western blot were used to analyze expression of FOXA1 in cervical cancer and adjacent tissue. The small-interfere RNA (siRNA) was adopted to down-regulate FOXA1 expression in HeLa cells. The effect of FOXA1 on apoptosis of HeLa cells was detected by using thiazolyl blue tetrazolium bromide (MTT) method. The apoptosis rate of HeLa cells was detected by using flow cytometry. The Western blot was selected to evaluate the epithelial mesenchymal transition (EMT) related protein, vimentin, E-cadherin, and vascular endothelial growth factor (VEGF) changes. RESULTS: Compared with adjacent tissues, FOXA1 mRNA and protein expressions significantly increased in cervical cancer (p<0.05). SiRNA significantly reduced FOXA1 expression in Hela cells compared with the control group and siRNA-NC group, thus inhibiting tumor cell proliferation and enhancing cell apoptosis rate (p<0.05). E-cadherin elevated, Vimentin decreased, and VEGF reduced after FOXA1 siRNA treatment. CONCLUSIONS: FOXA1 expression increased in cervical cancer. Inhibition of FOXA1 expression blocked the proliferation of cervical cancer, promoted tumor cell apoptosis, suppressed the occurrence of EMT and VEGF production, and can regulate cervical cancer metastasis. FOXA1 can be used as a new molecular biological target for cervical cancer diagnosis and treatment.

The cardiovascular macrophage: a missing link between gut microbiota and cardiovascular diseases?

The prevalence of cardiovascular diseases is on the rise. Interventions that would aid prevention or treatment of these diseases are essential. The microbes residing in the gut, collectively called "gut microbiota", produce a plethora of compounds that enter the bloodstream and affect the cardiovascular system. Signals ascending from gut microbiome are believed to modulate differentiation and functional activity of macrophages residing in perivascular tissue, atherosclerotic plaques, and perivascular areas of the brain. Cardiovascular macrophages may be the key players that transform the signals ascending from gut microbiome into increased predisposition to cardiovascular diseases. The present review summarizes the knowledge to date on potential relationships between gut microbiota, cardiovascular macrophages, and cardiovascular diseases.

Long non-coding RNA BANCR indicates poor prognosis for breast cancer and promotes cell proliferation and invasion.

OBJECTIVE: We investigated the expression of human long non-coding ribonucleic acid (lncRNA), BRAF-activated non-coding RNA (BANCR) in breast cancer tissues and its effects on the in vitro proliferation, apoptosis, invasion and metastasis of breast cancer cells; also, we investigated its possible mechanism. PATIENTS AND METHODS: The expressions of BANCR in 65 pairs of breast cancer tissues and para-carcinoma normal breast tissues were detected by Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). The expressions of BANCR in normal breast epithelial cells (MCF10A) and breast cancer cells (MCF-7, MDA-MB-231, SKBR3 and BT-20) were further detected. The knockdown efficiency of BANCR small interfering RNA (siRNA) in MCF-7 cells was detected by qRT-PCR. The effects of BANCR knockdown on proliferation, apoptosis, invasion and metastasis capacities of MCF-7 cells were explored via methyl thiazolyl tetrazolium (MTT) proliferation assay, cell colony formation assay, fluorescence-activated cell sorting (FACS) and transwell migration assay. Western blotting was used to detect the changes in expressions of apoptosis-related proteins, epithelial-mesenchymal transition (EMT)-related proteins and matrix metalloproteinases (MMPs) after knockdown of BANCR. RESULTS: The expression level of lncRNA BANCR in breast cancer tissues was significantly higher than that in para-carcinoma normal tissues. The prognosis of patients in low-expression BANCR group was significantly superior to that of patients in high-expression BANCR group. After BANCR knockdown in breast cancer MCF-7 cells, the cell proliferation and colony formation capacities were significantly inhibited. Further mechanism research showed that inhibiting BANCR could promote the apoptosis of MCF-7 cells. Results of Western blotting revealed that the expressions of B-cell lymphoma 2 associated X protein (BAX), cleaved-Caspase-3 and cleaved-poly adenosine diphosphate-ribose polymerase (PARP) in knockdown group were significantly up-regulated compared with those in control group. Both wound-healing assay and transwell migration assay showed that the down-regulation of lncRNA BANCR could inhibit the invasion and metastasis capacities of MCF-7 cells, whose mechanism was related to the inhibition of EMT process and down-regulation of MMP expressions in cells. CONCLUSIONS: LncRNA BANCR is highly expressed in breast cancer, which is significantly correlated with the prognosis of patients; moreover, it can promote the growth, invasion and metastasis of ovarian cancer cells. The down-regulation of BANCR can inhibit the proliferation, invasion and metastasis capacities of MCF-7 cells.

Adiponectin attenuates endoplasmic reticulum stress and alveolar epithelial apoptosis in COPD rats.

OBJECTIVE: The present study was designed to evaluate the effect of Adiponectin (APN) against alveolar epithelial apoptosis in chronic obstructive pulmonary disease (COPD) rat models. MATERIALS AND METHODS: Thirty-six male Sprague-Dawley (SD) rats were randomly assigned to three groups: Sham group, COPD group, and COPD + APN group (2.5 ug/kg/day). To assess the effect of APN, histopathological evaluations, lung function, and the apoptotic index (AI) of alveolar septal cells, were performed. In addition, the levels of oxidative stress and endoplasmic reticulum stress were measured. RESULTS: HE staining demonstrated that APN inhibited pathological injury in COPD rats. In addition, APN could restore the levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in serum. APN also inhibited the levels of endoplasmic reticulum stress pathway including CHOP, phospho-JNK and Caspase-12 in alveolar epithelial cell. Furthermore, APN significantly inhibited the protein levels of Caspase-3 and apoptosis in alveolar epithelial cell of COPD rats. CONCLUSIONS: Our findings suggested that APN might effectively ameliorate the progression of COPD via inhibiting the endoplasmic reticulum stress-induced alveolar epithelial apoptosis in rats.

Down-regulation of survivin enhances paclitaxel-induced Hela cell apoptosis.

OBJECTIVE: Paclitaxel is one of the common anticancer drugs in the treatment of cervical cancer, while the mechanism of restraining and killing cancer cells is still unclear. This study aimed to investigate the molecular mechanism of paclitaxel in regulating proliferation and apoptosis of cervical cancer Hela cells. MATERIALS AND METHODS: Paclitaxel at 2 µmol/L was used to treat Hela cells for 48 h. MTT assay and flow cytometry were applied to test Hela cells proliferation and apoptosis respectively. Western blot was adopted to determine the expression of survivin. SiRNA was performed to suppress survivin protein expression in Hela cells. RESULTS: Paclitaxel restrained Hela cells growth and induced apoptosis. Also, paclitaxel treatment significantly reduced survivin protein expression in Hela cells. Moreover, survivin siRNA transfection further promoted Hela cells apoptosis after intervention by 2 µmol/L paclitaxel. CONCLUSIONS: Down-regulation of survivin promoted paclitaxel-induced apoptosis of cervical cancer Hela cells.

Cultivation of Chlorella sp. with livestock waste compost for lipid production.

Cultivation of microalgae Chlorella sp. with livestock waste compost as an alternative nutrient source was investigated in this present study. Five culture media with different nutrient concentrations were prepared. The characteristics of algal growth and lipid production were examined. The results showed that the specific growth rate together with biomass and lipid productivities was different among all the cultures. As the initial nutrient concentration decreased, the lipid content of Chlorella sp. increased. The variations in lipid productivity of Chlorella sp. among all the cultures were mainly due to the deviations in biomass productivity. The livestock waste compost medium with 2000mgL-1COD provided an optimal nutrient concentration for Chlorella sp. cultivation, where the highest productivities of biomass (288.84mgL-1day-1) and lipid (104.89mgL-1day-1) were presented.

Biomechanical effects of interbody cage height on cervical spine / 医用生物力学

Objective To investigate the biomechanical effects of interbody cage height on cervical spine during anterior cervical discectomy and fusion (ACDF) surgery, so as to provide references for selection of interbody cage. Methods The finite element model of normal cervical spine (C2-7) was built and validated, and the cages with different height (5, 6, 7, 8 mm) were implanted into C5-6 disc (cage5, 6, 7, 8 model). All the models were loaded with pure moment of 1.5 N•m to produce flexion, extension, blending and axial torsion motions on the cervical spine, and the effects of cage height on range of motion (ROM), facet joint stress, intervertebral pressure in cervical spine were investigated. Results The intervertebral angle at the fusion segment increased by 0.68°with per 1 mm-increase of height. The ROM at C5-6 after cage implantation was less than 0.44°. The influence of cage height on ROM in C4-5 was greater than that in C6-7, and the changes of ROM in non-fusion segments were less than 7.3%. The cage height variation had a smaller impact on the facet joint stress and intervertebral pressure. The stresses in the capsular ligament, cage and screw-plate system increased gradually with the increase of cage height, and these stresses in the cage6, 7, 8 models were much higher than those in the cage5 model. Conclusions For patients who need implanting fusion cage, the cage height should be 0-1 mm greater than the original intervertebral space height.