MiR-513a-3p inhibits EMT mediated by HOXB7 and promotes sensitivity to cisplatin in ovarian cancer cells.

OBJECTIVE: Our aim was to investigate the biological function and mechanism of action of miR-513a-3p in ovarian cancer cells. MATERIALS AND METHODS: In this study, qRT-PCR, Western blots, and immunohistochemistry experiments were among the methods used to examine the expression of miR-513a-3p, HOXB7, and related transcripts within ovarian cancer cells. An MTT assay was conducted to evaluate the viability of ovarian cancer cells in the presence of cisplatin. Transwell and wound-healing assays were performed to examine cell migration and invasion. Dual-Luciferase reporter assays were used to evaluate interactions among the aforementioned target genes. In vivo tumorigenesis experiments were conducted to verify biological effects of miR-513a-3p and HOXB7. RESULTS: HOXB7 expression was relatively higher and MiR-513a-3p expression was relatively lower in ovarian cancer cells. Down-regulated expression of miR-513a-3p promoted cell movement via its ability to regulate epithelial-mesenchymal transition (EMT). Furthermore, decreased expression of miR-513a-3p resulted in increased sensitivity to cisplatin and resulted in poor prognosis in ovarian cancer patients who had relapsed after treatment with cisplatin. However, HOXB7 reversed the impact of miR-513a-3p in ovarian cancer cells. These results suggested that miR-513a-3p altered EMT mediated by HOXB7 and cisplatin-resistance. CONCLUSIONS: MiR-513a-3p plays a critical role in promoting sensitivity to cisplatin and tumorigenesis via targeting HOXB7 in ovarian cancer.

Significant association of DIRC1 overexpression with tumor progression and poor prognosis in gastric cancer.

OBJECTIVE: DIRC1, Disrupted in Renal Cancer 1, was identified as a breakpoint-spanning gene in a chromosomal translocation, which was associated with the onset and progression of some cancers. However, the expression in human gastric cancer (GC) and the role of DIRC1 in human gastric tumorigenesis are unknown. Thereby, the main purpose of this study was to unearth the association of DIRC1 with GC. MATERIALS AND METHODS: By analyzing The Cancer Genome Atlas (TCGA) datasets, the expression of DIRC1 in GC and normal gastric tissues were compared. Besides, its association with clinicopathological significance, overall survival (OS) and independent prognosis were analyzed by Pearson's x2 test, Kaplan-Meier method and Cox proportional hazards model, respectively. Functionally, the knockdown of DIRC1 was performed by siRNA method; moreover, its effects on the proliferation and metastasis of GC cells were examined by CCK-8 and transwell assays. Furthermore, the key markers of protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling were tested by Western blot assay. RESULTS: The results showed that high expression of DIRC1 was found in GC tissues compared with normal gastric tissues. High expression of DIRC1 was associated with more cases of severer tumor malignancy and shorter OS; besides, high-level of DIRC1 was suggested to be an independent prognostic factor for GC. Furthermore, the knockdown of DIRC1 inhibited SGC-7901 GC cells proliferation, migration and invasion. Mechanically, the activity of AKT/mTOR signaling was suppressed by the knockdown of DIRC1. CONCLUSIONS: These findings offer clinical associations and an in vitro evidence showing that the knockdown of DIRC1 impeded the GC carcinogenicity possibly via suppression of AKT/mTOR signaling. This work might provide a potential therapeutic target for GC treatment.

Liver injury induced by clomiphene citrate: A case report and literature reviews.

WHAT IS KNOWN AND OBJECTIVE: Clomiphene citrate is used to cause ovulation in females and to increase semen production in males. Clomiphene citrate is well tolerated in most patients and rarely induces liver injury. We report a case of liver injury which is associated with administration of clomiphene citrate in a male patient. CASE SUMMARY: A 31-year-old man who was treated by clomiphene citrate for 5 days was transferred to our emergency room with reddish-brown urine and upper abdominal pain. The levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were elevated. Based on the subsequent examination, he was diagnosed with liver injury and cholecystitis. The levels of AST and ALT returned to normal range after discontinuation of clomiphene citrate and symptomatic treatment. WHAT IS NEW AND CONCLUSION: The mechanism of liver injury caused by clomiphene citrate is still unclear. Polymorphism of CYP2D6 may have had an effect. Liver function tests should be performed when there is upper abdominal pain after administration of clomiphene citrate.

Polymorphisms of CASR gene increase the risk of primary hyperparathyroidism.

OBJECTIVE: To evaluate correlations between polymorphisms of calcium-sensing receptor (CASR) gene [A986S (rs1081725), R990G (rs1042636) and Q1011E (rs1801726)] and the risk of primary hyperparathyroidism (PHPT) among human population. METHODS: Relevant studies were retrieved from online databases using computer-based search strategies, which were then supplemented by manual search strategies. Case-control studies related to our topic were identified based on strict inclusion and exclusion criteria. Statistical analyses were conducted using the Comprehensive Meta-analysis 2.0 (Biostat Inc., Englewood, NJ, USA). RESULTS: We retrieved 202 studies from online databases and other sources initially and eventually enrolled six studies into our meta-analysis. These six studies contained a sum of 693 PHPT patients and 1252 healthy controls. Our meta-analysis results showed that single nucleotide polymorphisms (SNPs) of CASR gene A986S (rs1081725) and R990G (rs1042636), but not Q1011E (rs1801726), may increase the risk of PHPT [A986S (rs1081725): allele model: P = 0.013; dominant model: P = 0.044; R990G (rs1042636): allele model: P = 0.023; dominant model: P = 0.026)]. Subgroup analyses based on ethnicity showed that among Asians, A986S (rs1081725) increased the PHPT risk (P = 0.04) under the allele model, but not under the dominant model. Among Caucasians, there was no association between gene frequencies and PHPT under both the allele and dominant model. In Asians, no significant association was observed between R990G (rs1042636) and PHPT risk, but in Caucasians, R990G (rs1042636) significantly increased the incidence of PHPT [R990G (rs1042636): allele model: P = 0.015; dominant model: P = 0.009)]. CONCLUSION: Our results indicate that SNPs of CASR gene A986S (rs1081725) and R990G (rs1042636) may increase the risk of PHPT, and the polymorphisms can potentially be used as important biological markers for early diagnosis of PHPT.

Sam68 expression and cytoplasmic localization is correlated with lymph node metastasis as well as prognosis in patients with early-stage cervical cancer.

BACKGROUND: This study was aimed at investigating the role and molecular mechanism of Sam68 in cervical cancer lymph node metastasis. MATERIALS AND METHODS: Sam68 expression profile was detected by quantitative polymerase chain reaction, western blotting and immunohistochemical staining. Short hairpin RNA interfering approach was employed to suppress endogenous Sam68 expression in cervical cancer cells to determine its role in metastasis and the possible mechanism. RESULTS: Sam68 expression in cervical cancer was significantly up-regulated at both messenger RNA and protein levels compared with that in normal cervical tissues. The high expression level of Sam68 and its cytoplasmic localization were significantly associated with risk factors including pelvic lymph node metastasis (P < 0.001), and served as independent prognostic factors for predicting shortening of the overall survival time and disease-free survival time in patients with early-stage cervical cancer. Moreover, down-regulation of Sam68 in cervical cancer cells remarkably inhibited cellular motility and invasion. In addition, down-regulation of Sam68 reversed epithelial-mesenchymal transition through inhibiting the Akt/ GSK-3ß/Snail pathway. CONCLUSION: This study demonstrated that Sam68 could induce cervical cancer lymph node metastasis through regulating epithelial-mesenchymal transition, and Sam68 expression profile possessed the potential to serve as predictors of pelvic lymph node metastasis in cervical cancer patients.

Bacterial community structure and bamA gene diversity in anaerobic degradation of toluene and benzoate under denitrifying conditions.

AIM: To characterize the microbial community structure and bamA gene diversity involved in anaerobic degradation of toluene and benzoate under denitrifying conditions. METHODS AND RESULTS: Nitrate-reducing enrichment cultures were established on either toluene, benzoate or without additional substrate. Bacterial community structures were characterized by 16S rRNA gene-based PCR-DGGE analysis. bamA gene diversity was analysed using DGGE and cloning methods. The results showed that bamA gene related to bamA of Thauera chlorobenzoica was dominant in toluene and benzoate cultures. However, a greater diversity of sequences was obtained in benzoate cultures. Low diversity of bamA gene was observed, and some similarities of bamA were also found between active cultures and backgrounds, suggesting that potential natural attenuation of aromatic compounds might occur. CONCLUSIONS: The combined analysis of 16S rRNA and bamA genes suggests that the species related to genera Thauera dominated toluene- and benzoate-degrading cultures. The combination of multiple methods (DGGE and cloning) provides a more complete picture of bamA gene diversity. SIGNIFICANCE AND IMPACT OF THE STUDY: To our knowledge, this is the first report of bamA gene in denitrifying enrichments using DGGE and cloning analysis.

RU486 (mifepristone) ameliorates cognitive dysfunction and reverses the down-regulation of astrocytic N-myc downstream-regulated gene 2 in streptozotocin-induced type-1 diabetic rats.

Diabetic cognitive dysfunction (DCD), usually accompanied with chronically elevated glucocorticoids and hippocampal astrocytic alterations, is one of the most serious complications in patients with type-1 diabetes. However, the role for chronically elevated glucocorticoids and hippocampal astrocytic activations in DCD remains to be elucidated, and it is not clear whether astrocytic N-myc downstream-regulated gene 2 (NDRG2, involved in cell differentiation and development) participated in DCD. In the present study, three months after streptozotocin (STZ)-induced type-1 diabetes onset, rats showed cognitive impairments in Morris water maze test as well as elevated corticosterone level. Diabetic rats also presented down-regulation of glial fibrillary acidic protein (GFAP, a key indicator of astrocytic reactivity) and NDRG2 in hippocampus revealed by immunohistochemistry staining, real-time PCR and Western blot. Moreover, the diabetic cognitive impairments were ameliorated by 9-day glucocorticoids receptor (GR) blockade with RU486, and the down-regulation of hippocampal NDRG2 and GFAP in diabetic animals was also attenuated by 9-day GR blockade. These results suggest that glucocorticoids-GR system is crucial for DCD, and that astrocytic reactivity and NDRG2 are involved in these processes. Thus, inhibiting GR activation in the hippocampus may be a novel therapeutic strategy for treating DCD.

The anti-proliferative effects of recombinant human lysozyme on human gastric cancer cells.

We investigated the anti-proliferative effects of recombinant human lysozyme (rHlys) on gastric cancer cell lines and normal human lung fibroblasts. Using conventional molecular cloning techniques we purified rHlys, which we incubated with cultured cells and measured the effects on cell proliferation and viability. At concentrations of 100 and 1000 microg/l, rHlys significantly inhibited the growth of human gastric cancer cell lines. In contrast, 10 and 50 microg/l of rHlys stimulated gastric cancer cell growth. None of the concentrations of rHlys affected cell viability. Only the highest concentration of rHlys (1000 microg/l) inhibited human lung fibroblast growth. Our results suggest that 100 microg/l is the optimum growth inhibiting concentration, which inhibited cancer cell growth but not normal cell growth. Our in vitro findings suggest that genetically engineered rHlys might inhibit human gastric cancer cell proliferation in vivo, so it might warrant further investigation as a potential novel anti-cancer agent.

Cimifoetisides VI and VII Two new cyclolanostanol triterpene glycosides from the aerial parts of Cimicifuga foetida.

Two new cyclolanostanol triterpene glycosides, cimifoetiside VI (1) and cimifoetiside VII (2), and one known compound were isolated from the aerial parts of Cimicifuga foetida L. On the basis of spectral and chemical evidences, the structures of 1 and 2 were elucidated to be (23R,24S)-24-O-acetylisodahurinol-3-O-beta-d-galactopyranoside and (23R,24R)-24-O-acetylshengmanol-3-O-beta-d-glucopyranosyl-(1'' --> 2')-beta-d-xylopyranoside. The known compound was identified as (23R,24R)-24-O-acetylshengmanol-3-O-beta-d-galactopyranoside (3).