Silencing IKBKE inhibits the migration and invasion of glioblastoma by promoting Snail1 degradation.

PURPOSE: Glioblastoma multiforme (GBM) is one of the most common malignant brain tumors in adults and has high mortality and relapse rates. Over the past few years, great advances have been made in the diagnosis and treatment of GBM, but unfortunately, the five-year overall survival rate of GBM patients is approximately 5.1%. Inhibitor of nuclear factor kappa-B kinase subunit epsilon (IKBKE) is a major oncogenic protein in tumors and can promote evil development of GBM. Snail1, a key inducer of the epithelial-mesenchymal transition (EMT) transcription factor, is subjected to ubiquitination and degradation, but the mechanism by which Snail1 is stabilized in tumors remains unclear. Our study aimed to investigate the mechanism of IKBKE regulating Snail1 in GBM. METHODS: First, we analyzed the correlation between the expression of IKBKE and the tumor grade and prognosis through public databases and laboratory specimen libraries. Second, immunohistochemistry (IHC) and western blot were used to detect the correlation between IKBKE and Snail expression in glioma samples and cell lines. Western blot and immunofluorescence (IF) experiments were used to detect the quality and distribution of IKBKE and Snail1 proteins. Third, In situ animal model of intracranial glioma to detect the regulatory effect of IKBKE on intracranial tumors. RESULTS: In this study, Our study reveals a new connection between IKBKE and Snail1, where IKBKE can directly bind to Snail1, translocate Snail1 into the nucleus from the cytoplasm. Downregulation of IKBKE results in Snail1 destabilization and impairs the tumor cell migration and invasion capabilities. CONCLUSION: Our studies suggest that the IKBKE-Snail1 axis may serve as a potential therapeutic target for GBM treatment.

Circulating exosomal miRNAs and cancer early diagnosis.

Microribonucleic acids (miRNAs) are small non-coding ribonucleic acids (ncRNAs), which can affect recognition of homologous sequences and interfere with transcription. It plays key roles in the initiation, development, resistance, metastasis or recurrence of cancers. Identifying circulatory indicators will positively improve the prognosis and quality of life of patients with early cancer. Previous studies have shown that miRNA is highly involved in cancer. In addition, miRNA derived from cancers can be encapsulated as exosomes and further extracted into circulatory systems to realize malignant functions. It indicates that circulating exosome-derived miRNAs have the potential to replace conventional biomarkers as cancer derived exosomes carrying miRNAs can be identified by specific markers and might be more stable and accurate for early diagnosis.

COPB2: a transport protein with multifaceted roles in cancer development and progression.

The Coatomer protein complex subunit beta 2 (COPB2) is involved in the formation of the COPI coatomer protein complex and is responsible for the transport of vesicles between the Golgi apparatus and the endoplasmic reticulum. It plays an important role in maintaining the integrity of these cellular organelles, as well as in maintaining cell homeostasis. More importantly, COPB2 plays key roles in embryonic development and tumor progression. COPB2 is regarded as a vital oncogene in several cancer types and has been implicated in tumor cell proliferation, survival, invasion, and metastasis. Here, we summarize the current knowledge on the roles of COPB2 in cancer development and progression in the context of the hallmarks of cancer.

Elevated expression of GNAS promotes breast cancer cell proliferation and migration via the PI3K/AKT/Snail1/E-cadherin axis.

PURPOSE: Although it has been well established that G protein plays pivotal roles in physiologic or pathologic conditions, including cancer formation, its role in breast cancer, especially specific subunits, remains largely unknown. Our work aimed to evaluate the correlation of the G protein alpha subunit (GNAS) with breast cancer and to investigate the underlying molecular mechanism. METHODS: The expression of GNAS was determined by breast tumor tissue microarray of 150 patients with complete follow-up information. The correlation between GNAS expression and clinical features was assessed. CCK8, EdU incorporation, flow cytometry, wound healing, transwell, western blot and tumor formation assays were carried out in nude mice to study the biological function of GNAS and the underlying molecular mechanism in breast cancer by silencing GNAS using a specific siRNA. RESULTS: High GNAS expression showed a close correlation with a reduced overall survival (p = 0.021), frequent distal metastasis (p = 0.026), advanced clinical stage (p = 0.001), stronger cell proliferation (ki67+ positive cell rate, p = 0.0351) and enhanced cancer cell migration, which was further confirmed by in vitro and in vivo assays and might be dependent on the PI3K/AKT/Snail1/E-cadherin axis. CONCLUSION: The data suggested that GNAS promoted breast cancer cell proliferation and migration (EMT) through the PI3K/AKT/Snail1/E-cadherin signaling pathway. These findings also indicate that GNAS can serve as a potential prognostic indicator and novel therapeutic target in breast cancer.

Letrozole improves the sensitivity of breast cancer cells overexpressing aromatase to cisplatin via down-regulation of FEN1.

PURPOSE: Flap endonuclease 1 (FEN1) is up-regulated by estrogen (17ß-estradiol, E2) and related to cisplatin resistance of human breast cancer cells. Letrozole, an aromatase inhibitor, suppresses the change of testosterone into estrogen and is frequently used to treat breast cancer. However, the effects of letrozole on FEN1 expression and cisplatin sensitivity in breast cancer cells overexpressing aromatase have not been revealed. METHODS: The expression of FEN1 and the proteins in ERK/Elk-1 signaling were evaluated by RT-PCR and Western blot. Cisplatin sensitivity was explored through CCK-8 and flow cytometry analysis, respectively. FEN1 siRNAs and FEN1 expression plasmid were transfected into cells to down-regulate or up-regulate FEN1 expression. The promotor activity of FEN1 was detected using luciferase reporter assay. RESULTS: FEN1 down-regulation improved cisplatin sensitivity of breast cancer cells overexpressing aromatase. Letrozole down-regulated FEN1 expression and increased cisplatin sensitivity. The sensitizing effect of letrozole to cisplatin was dependent on FEN1 down-regulation. FEN1 overexpression could block the sensitizing effect of letrozole to cisplatin. Testosterone up-regulated the promotor activity, protein expression of FEN1, and phosphorylation of ERK/Elk-1, which could be eliminated by both letrozole and MEK1/2 inhibitor U0126. Letrozole down-regulated FEN1 expression in an ERK/Elk-1-dependent manner. CONCLUSIONS: Our findings clearly demonstrate that letrozole improves cisplatin sensitivity of breast cancer cells overexpressing aromatase via down-regulation of FEN1 and suggest that a combined use of letrozole and cisplatin may be a potential treatment protocol for relieving cisplatin resistance in human breast cancer.

Evidence for supercoughers in an analysis of six tuberculosis cohorts from China, Peru, The Gambia and Uganda.

BACKGROUND: It is very difficult to observe tuberculosis (TB) transmission chains and thus, identify superspreaders. We investigate cough duration as a proxy measure of transmission to assess the presence of potential TB superspreaders.DESIGN: We analyzed six studies from China, Peru, The Gambia and Uganda, and determined the distribution of cough duration and compared it with several theoretical distributions. To determine factors associated with cough duration, we used linear regression and boosted regression trees to examine the predictive power of patient, clinical and environmental characteristics.RESULTS: We found within-study heterogeneity in cough duration and strong similarities across studies. Approximately 20% of patients contributed 50% of total cough days, and around 50% of patients contributed 80% of total cough days. The cough duration distribution suggested an initially increasing, and subsequently, decreasing hazard of diagnosis. While some of the exposure variables showed statistically significant associations with cough duration, none of them had a strong effect. Multivariate analyses of different model types did not produce a model that had good predictive power.CONCLUSION: We found consistent evidence for the presence of supercoughers, but no characteristics predictive of such individuals.

Circular RNAs and exosomes in cancer: a mysterious connection.

Circular RNAs (CircRNAs) are a type of non-coding RNAs (NcRNAs) with a closed annular structure. Until next-generation sequencing (NGS) is developed, the misunderstanding of circRNAs 'splicing error' has changed, and the mysterious veil of circRNAs has been revealed. NGS provides an approach to investigate circRNAs. Many scholars point out that circRNAs may play an important role in many diseases, especially cancer. At the same time, exosomes, as a kind of extracellular vesicles loaded with many contents, are a hotspot in recent years. They can act as 'messengers' between cells, especially in cancer. Lately, it is interesting circRNAs are enriched and stable in exosomes, also called exo-circRNAs, and there have been several articles on circRNAs associated with exosomes. In this review, we summarize the characteristics of circRNAs, especially its main functions. Then, we briefly introduce exosomes and their function in cancer. Finally, the known relation between circRNAs and exosomes is discussed. With further researches, exo-circRNAs may be a novel pathway for cancer diagnosis and targeted therapy.

Application of EGFR inhibitor reduces circulating tumor cells during transcatheter arterial embolization.

PURPOSE: Transcatheter arterial embolization (TAE) has been widely used in treating non-curative hepatocellular carcinoma (HCC). However, it is noticed that TAE may cause invasion of some cancer cells into circulation, resulting in distal metastasis and poor therapeutic outcome. Here, we aimed to reduce the side effects of TAE using the inhibitors for epidermal growth factor receptor (EGFR). METHODS: Transient hepatic artery ligation (HAL) was used as a mouse model for TAE. EGFR inhibitors were applied. Tumor size, presence of tumor cells in circulation, distal tumor formation, and activation of genes associated with tumor cell invasion and metastasis were analyzed. RESULTS: Inhibitors for EGFR significantly reduced the size of primary tumor, presence of tumor cells in circulation, and distal tumor formation after HAL. Further studies showed that EGFR inhibition suppressed several genes associated with tumor cell invasion and metastasis, such as vascular endothelial growth factor-A, stromal cell-derived factor 1, and Slug. CONCLUSION: EGFR inhibitor application may reduce circulating cancer cells during TAE and thus improve the therapy for advanced HCC.

Two novel Mg(II)-based and Zn(II)-based complexes: inhibiting growth of human liver cancer cells

Two new Mg(II)-based and Zn(II)-based coordination polymers, {[Mg3(BTB)(DMA)4](DMA)2}n (1, H3BTB=1,3,5-benzenetrisbenzoic acid, DMA=N,N-dimethylacetamide) and {(H2NMe2)2[Zn3(BTB)2(OH)(Im)](DMF)9(MeOH)7}n (2, Im=imidazole, DMF=N,N-dimethylformamide), have been successfully synthesized and structurally characterized under solvothermal conditions. 1 contains a linear [Mg3(COO)6] cluster that connected by the fully deprotonated BTB3- ligands to give a kgd-type 2D bilayer structure; 2 represents a microporous 3D pillar-layered system based on the binuclear Zn units and pillared Im ligands, which shows a (3,5)-connected hms topological net. In addition, in vitro anticancer activities of compounds 1 and 2 on 4 human liver cancer cells (HB611, HHCC, BEL-7405 and SMMC-7721) were determined.

Two novel Mg(II)-based and Zn(II)-based complexes: inhibiting growth of human liver cancer cells.

Two new Mg(II)-based and Zn(II)-based coordination polymers, {[Mg3(BTB)(DMA)4](DMA)2}n (1, H3BTB=1,3,5-benzenetrisbenzoic acid, DMA=N,N-dimethylacetamide) and {(H2NMe2)2[Zn3(BTB)2(OH)(Im)](DMF)9(MeOH)7}n (2, Im=imidazole, DMF=N,N-dimethylformamide), have been successfully synthesized and structurally characterized under solvothermal conditions. 1 contains a linear [Mg3(COO)6] cluster that connected by the fully deprotonated BTB3- ligands to give a kgd-type 2D bilayer structure; 2 represents a microporous 3D pillar-layered system based on the binuclear Zn units and pillared Im ligands, which shows a (3,5)-connected hms topological net. In addition, in vitro anticancer activities of compounds 1 and 2 on 4 human liver cancer cells (HB611, HHCC, BEL-7405 and SMMC-7721) were determined.

Mechanism of reversal of multidrug resistance by curcumin in human colorectal cancer cell line HCT-8/5-FU.

Since there are no studies on the reversal of multidrug resistance by curcumin in the human colorectal cancer cell line HCT-8/5-FU, our aim was to search for highly efficient reversal agents and investigate the underlying mechanisms of this reversal. The cytotoxic effects of curcumin and 5-FU on HCT-8 and HCT-8/5-FU cells and the reversal effects of 5-FU in combination with curcumin on HCT-8/5-FU cells were measured using cell counting kit-8. Apoptosis and the cell cycle were analyzed by flow cytometry. Protein and mRNA expression levels of BCL-2, survivin, P-gp, and HSP-27 were detected by western blotting and quantitative real-time reverse transcription polymerase chain reaction, respectively. Curcumin inhibited the growth of HCT-8 and HCT-8/5-FU cells. It significantly reduced the IC50 of 5-FU for HCT-8/5-FU cells (P < 0.01) and the expression of BCL-2, survivin, P-gp, and HSP-27 in the cells. Curcumin can effectively reverse multidrug resistance in human colorectal cancer drug-resistant HCT-8/5-FU cells. The mechanism through which this occurs may be associated with decreased expression of BCL-2, survivin, P-gp, and HSP-27. Curcumin may therefore have clinical implications as a new agent for colorectal cancer.

Paracrine effect of bone marrow mesenchymal stem cells on proliferation, apoptosis, and alpha-actin-2 expression in hepatic stellate cells.

We investigated the paracrine effects of bone marrow mesenchymal stem cells (BMSCs) on the proliferation, apoptosis, and alpha-actin-2 (ACTA2) expression of hepatic stellate cells (HSCs), and explored the possible mechanisms of hepatocyte growth factor (HGF). We established a co-culture system by culturing BMSCs on the upper layer and HSCs on the lower layer of a 6-well Transwell plate. Normal HSCs were cultured alone as a control. Cell apoptosis was determined by flow cytometry. We detected the expression of ACTA2 mRNA and ACTA2 protein in HSC using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blotting, respectively. ACTA2 in HSCs was detected by fluorescent staining, and HGF in the co-culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA). The apoptotic rate of HSCs in the experiment group was 2.6 times that in the control group (P < 0.05). The expression levels of ACTA2 mRNA and ACTA2 protein were significantly inhibited in HSCs compared with the control group (P < 0.05). HGF concentration in the co-culture supernatant was 0.43 ± 0.47 mM in the experimental group, which was significantly higher than in the control group (0.16 ± 0.43 mM) (P < 0.05). The paracrine effect of BMSCs, which was caused by the suppression of ACTA2 and HGF expression, induced HSC apoptosis.

Isolation and Probiotic Potential of Lactobacillus Salivarius and Pediococcus Pentosaceus in Specific Pathogen Free Chickens

This study was conducted to isolate Lactobacillus salivarius and Pediococcus pentosaceus strains from cecal content and investigate their probiotic potential in specific pathogen free (SPF) chickens. L. salivarius and P. pentosaceus strains were isolated from the cecal content of SPF chickens and identified by 16s rDNA sequence analysis by BLAST analysis at the National Center for Biotechnology Information and phylogenetic analysis using DNAStar software. In an in vivo experiment, 180 7-day-old SPF chickens were randomly assigned into three groups. Group 1 served as a control that was fed a basal diet without probiotic supplementation, and groups 2 and 3 were fed the basal diets supplemented with L. salivarius and P. pentosaceus at 2×108 CFU/g, respectively. Body weight (BW), average daily gain (ADG), feed conversion ratio (FCR), dressing percentage (DP), and the apparent digestibility of crude protein (AD-CP) were calculated. We also determined meat color, fat content, shear force, water content and pH value of breast and thigh muscles; ammonia, urea nitrogen and uric acid content in plasma; fecal ammonia emission level and pH value; and Lactobacillus and Escherichia coli in ceca. Compared with the control group, L. salivarius and P. pentosaceus supplementation significantly increased BW, ADG, DP, AD-CP, fat content of meat, and the number of Lactobacillus in ceca (p 0.05), and decreased FCR, plasma ammonia content, fecal ammonia emission, and pH value and the number of E. coli in ceca (p 0.05). In the in vitro experiment, L. salivarius and P. pentosaceus treatments significantly decreased the ammonia content in medium compared with the control group without probiotic treatment (p 0.05). These results suggest that P. pentosaceus and L. salivarius strains show promising probiotic properties for improving growth, meat quality and microenvironment in chickens and decreasing ammonia content in the medium.

Isolation and Probiotic Potential of Lactobacillus Salivarius and Pediococcus Pentosaceus in Specific Pathogen Free Chickens

This study was conducted to isolate Lactobacillus salivarius and Pediococcus pentosaceus strains from cecal content and investigate their probiotic potential in specific pathogen free (SPF) chickens. L. salivarius and P. pentosaceus strains were isolated from the cecal content of SPF chickens and identified by 16s rDNA sequence analysis by BLAST analysis at the National Center for Biotechnology Information and phylogenetic analysis using DNAStar software. In an in vivo experiment, 180 7-day-old SPF chickens were randomly assigned into three groups. Group 1 served as a control that was fed a basal diet without probiotic supplementation, and groups 2 and 3 were fed the basal diets supplemented with L. salivarius and P. pentosaceus at 2×108 CFU/g, respectively. Body weight (BW), average daily gain (ADG), feed conversion ratio (FCR), dressing percentage (DP), and the apparent digestibility of crude protein (AD-CP) were calculated. We also determined meat color, fat content, shear force, water content and pH value of breast and thigh muscles; ammonia, urea nitrogen and uric acid content in plasma; fecal ammonia emission level and pH value; and Lactobacillus and Escherichia coli in ceca. Compared with the control group, L. salivarius and P. pentosaceus supplementation significantly increased BW, ADG, DP, AD-CP, fat content of meat, and the number of Lactobacillus in ceca (p 0.05), and decreased FCR, plasma ammonia content, fecal ammonia emission, and pH value and the number of E. coli in ceca (p 0.05). In the in vitro experiment, L. salivarius and P. pentosaceus treatments significantly decreased the ammonia content in medium compared with the control group without probiotic treatment (p 0.05). These results suggest that P. pentosaceus and L. salivarius strains show promising probiotic properties for improving growth, meat quality and microenvironment in chickens and decreasing ammonia content in the medium.(AU)

Differential expression of Toll-like receptors in goat dominant and nondominant follicles.

The mechanism of dominant follicle selection is unclear because of its physiological complexity. However, some studies have reported that the immune system plays an important role in reproductive physiology. The objective of the current study was to investigate the differential expression of Toll-like receptors (TLRs) in the dominant (DFs) and nondominant follicles (NFs), and to determine the correlation between the expression of TLRs and the related genes, such as WNT4 and FOXL2. In this comparative study, the expression levels of TLRs, WNT4, and FOXL2 genes of DFs and NFs were obtained from three Dazu black goats were estimated using the real-time PCR. Our results showed no significant difference in the expression of seven TLRs (excluding TLR2, TLR5, and TLR8), WNT4, and FOXL2 between the DFs and NFs. In addition, the mRNA expression levels of WNT4 significantly correlated with the relative expression of TLR6 (r = 0.949739, P < 0.01); however, no significant expression of the TLR genes was found to be associated with FOXL2 mRNA expression. Our results support the fact that TLRs are not involved in the process of dominant follicle selection; however, TLR6 might play a role in the development of follicles by interacting with WNT4.

Analyzing gene expression profiles in dilated cardiomyopathy via bioinformatics methods.

Dilated cardiomyopathy (DCM) is characterized by ventricular dilatation, and it is a common cause of heart failure and cardiac transplantation. This study aimed to explore potential DCM-related genes and their underlying regulatory mechanism using methods of bioinformatics. The gene expression profiles of GSE3586 were downloaded from Gene Expression Omnibus database, including 15 normal samples and 13 DCM samples. The differentially expressed genes (DEGs) were identified between normal and DCM samples using Limma package in R language. Pathway enrichment analysis of DEGs was then performed. Meanwhile, the potential transcription factors (TFs) and microRNAs (miRNAs) of these DEGs were predicted based on their binding sequences. In addition, DEGs were mapped to the cMap database to find the potential small molecule drugs. A total of 4777 genes were identified as DEGs by comparing gene expression profiles between DCM and control samples. DEGs were significantly enriched in 26 pathways, such as lymphocyte TarBase pathway and androgen receptor signaling pathway. Furthermore, potential TFs (SP1, LEF1, and NFAT) were identified, as well as potential miRNAs (miR-9, miR-200 family, and miR-30 family). Additionally, small molecules like isoflupredone and trihexyphenidyl were found to be potential therapeutic drugs for DCM. The identified DEGs (PRSS12 and FOXG1), potential TFs, as well as potential miRNAs, might be involved in DCM.

Relationship between genetic polymorphisms of methylenetetrahydrofolate reductase and breast cancer chemotherapy response.

Activity of methylenetetrahydrofolate reductase (MTHFR), an enzyme involved in folate metabolism, is influenced by mutations in the corresponding gene, contributing to a decrease in 5,10-MTHF. Due to such polymorphisms, individuals differ in MTHFR enzyme activity and plasma folate levels. We investigated the relationship between two common MTHFR polymorphisms (C677T and A1298C) and breast cancer (BC) chemotherapy response. From February 2013 to January 2016, 148 advanced BC patients at the Center Hospital of Cangzhou were enrolled and treated with six different chemotherapy regimens. Subjects were genotyped using polymerase chain reaction-restriction fragment length polymorphism. Forty-one (27.7%), 70 (47.3%), and 37 (25.0%) patients carried the C/C, C/T, and T/T C677T genotypes, respectively; 101 (68.2%), 42 (28.4%), and 5 (3.4%) had the A/A, A/C, and C/C genotypes of A1298C, respectively. Total chemotherapy efficacy was 66.9% (99/148), with 7 (4.7%), 92 (62.2%), 36 (24.3%), and 13 (8.8%) cases showing complete response, partial response, no change, and progressive disease, respectively. Chemotherapy regimens did not differ in effectiveness (P > 0.05). Efficacy rates associated with C677T C/C, C/T, and T/T genotypes were 58.5, 58.6, and 91.9%, respectively, with T/T carriers exhibiting significantly better responses than the C/C (P < 0.05) and C/T groups (P < 0.05). Effectiveness among A1298C A/A, A/C, and C/C carriers was 70.6, 64.3, and 0.0%, respectively, but no difference was established between these genotypes in this regard (P > 0.05). The MTHFR C677T genotype may be associated with BC chemotherapy response, and could be of great value in guiding individualized treatment for this disease.

Relationship between the polymorphisms in KCNQ1 and type 2 diabetes in Chinese Kazakh population.

Recently genome-wide association studies on East Asian populations reported an association between diabetes and several single nucleotide polymorphisms (SNPs) in a 40-kb linkage disequilibrium block in intron 15 of KCNQ1. However, the association between KCNQ1 variants and type 2 diabetes mellitus (T2DM) in Chinese Kazakh populations is unknown. We investigated the relationship between rs2237892 and rs2237895 SNPs in KCNQ1 and susceptibility to and clinical characteristics of T2DM in 100 Chinese Kazakh T2DM subjects and 100 healthy subjects. SNPs were genotyped by polymerase chain reaction-restriction fragment length polymorphism and the main anthropometric and biochemical parameters of individuals were assessed in the genotype groups (rs2237892: CC, CT, or TT, and rs2237895: AA, AC, or CC). Genotype distribution and allele frequencies of these two SNPs were not significantly different between T2DM and control groups (P > 0.05). The frequencies of CT and TT genotypes and T allele for the rs2237892 SNP in females with T2DM were significantly higher than that in the control group (genotype: P = 0.016, allele: P = 0.004). However, there were no significant differences among individuals with different genotypes with respect to the rs2237895 SNP (P > 0.05). The main anthropometric and biochemical parameters did not correlate with the rs2237892 or rs2237895 SNPs in the T2DM group (P > 0.05). Thus, the T allele-containing genotypes of the rs2237892 SNP in KCNQ1 may increase the susceptibility to T2DM in female Chinese Kazakh individuals, whereas the rs2237895 SNP may not be associated with T2DM in the Chinese Kazakh population.

Application of retrograde dissection method for isolation of bone marrow cells from rat femurs and tibiae.

Currently, there is no practical and efficient method for the isolation of bone marrow cells (BMCs) from rat femurs and tibiae. Here, we attempted to develop a rapid, simple, effective, and non-contaminating method for the isolation of BMCs from rat femurs and tibiae. Rat femurs and tibiae were dissected from the ankle to the hip joint; subsequently, a three-step "locate-slide-twist" procedure was performed using scissors and forceps to remove the femurs and tibiae completely, from the surrounding musculature. The bones were flushed with phosphate-buffered saline to harvest BMCs. The femurs and tibiae were dissected in 1.8 ± 0.6 min, and the BMC suspension preparation time was 13.1 ± 2.3 min. The bone marrow cavities did not incur any fractures or injuries during the isolation. Culture of harvested BMCs for 72 h led to a significant increase in cell number from 4.4 ± 0.3 x 106 to 6.9 ± 0.7 x 10(6) (P < 0.01) with no significant decrease in viability (98.1 ± 0.6% vs 96.2 ± 1.1%; P > 0.05). Microscopic examination of the isolated BMCs after the 72-h incubation period revealed the no-microbial or muscle cell contamination. Furthermore, flow cytometry revealed that cultured BMCs (72-h culture) grew well. Here, we have reported a rapid, simple, effective, and non-contaminating method for the isolation of BMCs from rat femurs and tibiae by using retrograde dissection. This method can be used to harvest a large number of viable BMCs without the risk of contamination from muscle and connective tissues.

Association between adiponectin gene T45G polymorphism and nonalcoholic fatty liver disease risk: a meta-analysis.

Numerous epidemiological investigations have evaluated the association between adiponectin gene T45G polymorphism and risk of nonalcoholic fatty liver disease (NAFLD). However, the results of these studies have proven to be inconsistent. Therefore, we conducted a meta-analysis to obtain a more accurate estimation of this association. Published articles were retrieved from PubMed and Web of Science databases and pooled odds ratios (ORs) with 95% confidence intervals (CIs) were calculated using fixed- or random-effect models. Five case-control studies incorporating 597 cases and 701 controls were included in this meta-analysis. No association between adiponectin gene T45G polymorphism and NAFLD was established (TT vs GG: OR = 0.83, 95%CI = 0.37-1.86; TG vs GG: OR = 0.76, 95%CI = 0.33-1.79; dominant model: OR = 0.83, 95%CI = 0.37-1.84; recessive model: OR = 1.10, 95%CI = 0.69-1.76). Moreover, in a subgroup analysis, no significant correlation was found among Asian subjects. In conclusion, the T45G polymorphism of the adiponectin gene may not constitute an NAFLD risk factor. However, this needs to be further validated in single large well-designed future studies.