HMGB1/RAGE axis mediates the apoptosis, invasion, autophagy, and angiogenesis of the renal cell carcinoma.

BACKGROUND: High mobility group box 1 protein (HMGB1) is a sort of non-histone protein in chromatin, which plays an important role in tumor proliferation, invasion, and immune escape. HMGB1-RAGE (receptor for advanced glycation end products) interactions have been reported to be important in a number of cancers. METHODS: CCK8, flow cytometry and qRT-PCR were used to detected cell viability, apoptosis and gene expression, respectively. RESULTS: In the present study, we demonstrated that HMGB1/RAGE axis regulated the cell proliferation, apoptosis, and invasion of the renal cell carcinoma (RCC). Further, we discovered that HMGB1/RAGE axis increased the expression of autophagic proteins LC3 and Beclin-1 in RCC. Finally, we used a coculture model of human umbilical vein endothelial cells with RCC cell lines to find out that HMGB1 also increased the expression of VEGF and VEGFR2 in human umbilical vein endothelial cells. An in vivo study further confirmed that HMGB1 knockdown inhibited RCC tumor growth. CONCLUSION: Our results illustrated that HMGB1/RAGE axis mediated RCC cell viability, apoptosis, invasion, autophagy, and angiogenesis, which provides a novel theoretical basis for using HMGB1 as the target in RCC.

[The role of sub-transform of macrophages in renal ischemia/reperfusion injury in rats].

OBJECTIVE: To investigate the role of sub-transform macrophage in ischemia/reperfusion renal injury in rats, as well as under-lying mechanisms. METHODS: Thirty male Sprague-Dawley rats were randomly divided into ischemia/reperfusion (IRI, n=24, renal artery was occluded for 45 min) group and sham-operation (Sham, n=6) group. The kidneys in IRI group were collected at 0, 6, 24 and 72 h after operation (6 rats for each time point). The injury of the kidney was detected with HE staining. Immunohistochemistry staining was performed to evaluate the expression of proliferating cell nuclear antigen (PCNA). Real-time PCR was used to detect the mRNA expression of macrophage migration inhibitory factor (MIF). Moreover, the expression and location of MIF, monocyte chemoattractant protein-1 (MCP-1) and macrophage marker CD68 were examined by immunofluorescence staining. Most importantly, the distribution of macrophage subtypes M1 and M2 was analyzed by flow cytometry. RESULTS: The worst pathologic damage of the renal tissues, as well as infiltration of inflammatory cells, was observed at 24 h after operation in IRI rats, with obvious recovery afterwards. Immunohistochemistry staining showed that the expression of PCNA was significantly increased after the ischemia/reperfusion, peaking at 6 h and reducing at 72 h after operation. Compared with sham group, the levels of MIF at mRNA and protein levels were both significantly increased after the ischemia/reperfusion, while the expression of MCP-1 was peaked at 6 h and decreased afterwards. Moreover, the expression of CD68-positive macrophages were significantly increased in IRI rats, with peaking at 24 h and reducing at 72 h. Furthermore, after 6 h of reperfusion, the percentage of M1 macrophages reached the peak, and thereafter the relative expression of M1 and M2 was reduced and increased, respectively. CONCLUSIONS: In the early phase of ischemia/per-fu sion renal injury, M1 macrophage results in renal damage, and afterwards the M2 macrophage is increased and repairs the renal damage by improving the cell proliferation.

Simultaneous monitoring of CMV and BKV by quantitative PCR in renal transplant recipients.

Polyomavirus (BKV) and cytomegalovirus (CMV) are associated with renal graft failure. The aim was to establish a quantitative PCR method (Q-PCR) to detect BKV and CMV simultaneously. The conserved sequences of BKV and CMV were amplified and cloned into the plasmids as standards. The sensitivity, specificity and the precision of the assay were evaluated. Q-PCR was used to detect BKV and CMV DNA simultaneously in 480 blood samples of renal transplantation recipients. The sensitivity of the Q-PCR assay to detect BKV or CMV DNA reached 5×10(3)copies/mL. The use of control DNA verified that the assay could specifically detect the target DNA. The precision of the assay to quantify target DNA copies was acceptable (ICV 3.44% for BKV and 2.23% for CMV; differences between batches ICV 4.98% for BKV and 3.76% for CMV). In 480 samples, 130 samples (27.08%) were CMV DNA positive, which was significantly higher than the 64 BKV DNA positive samples (13.33%, p<0.05). BKV or CMV DNA positivity was significantly associated with high concentrations of Tacrolimus (TAC) (p value<0.05). The Q-PCR assay to detect both CMV and BKV DNA simultaneously was developed successfully with high sensitivity, precision, and time-effectiveness for clinical measurement.

Treatment with ginkgo biloba extract protects rats against acute pancreatitis-associated lung injury by modulating alveolar macrophage.

INTRODUCTION: Acute pancreatitis (AP) protease release induces lung parenchymal destruction via inflammatory mediators. Ginkgo biloba has been reported to have anti-inflammatory effects. AIM: To evaluate the effect of ginkgo biloba extract on experimental acute pancreatitis-associated lung injury in the rat and to investigate the underlying mechanisms. MATERIAL AND METHODS: Acute pancreatitis was induced in rats by injection of 5% sodium taurocholate into the biliary pancreatic duct. Ginkgo biloba extract (GBE) was administered and pancreas and lung injury were assessed by histological examination. Alveolar macrophages were harvested by bronchoalveolar lavage. Specificity fluorescent probe DAF-FM-DA was applied to observe nitric oxide (NO) bioavailability in alveolar macrophage. The expression of tumour necrosis factor α (TNF-α) and macrophage migration inhibitory factor (MIF) protein in alveolar macrophage was studied by ELISA. RESULTS: In sodium taurocholate-induced acute pancreatitis, treatment with GBE significantly protected rats against lung injury associated with pancreatitis in histological sections. Ginkgo biloba extract had a tendency to down-regulate NO bioavailability compared with the AP group, but without statistical significance. Moreover, TNF-α and MIF at protein levels in alveolar macrophage with GBE treatment were decreased compared with the AP group. CONCLUSIONS: These results suggest that GBE could effectively protect rats against acute pancreatitis-associated lung injury. The GBE may inhibit excessive activation of alveolar macrophages from acute pancreatitis-associated lung injury through down-regulation of generation of NO, TNF-α and MIF. These findings suggest that ginkgo biloba extract is a suitable candidate as an effective strategy against acute pancreatitis-associated lung injury.

The progression of the tubulointerstitial fibrosis driven by stress-induced "proliferation-death" vicious circle.

Several hypotheses have been developed to interpret the progression of tubulointerstitial fibrosis (TF), including senescence, epithelial-mesenchymal transition, inflammation, chronic hypoxia, and reactive oxygen species. All of these hypotheses are based on persistent cell injury and localized cell death. Proliferation of neighboring renal tubular epithelial cells (RTECs) is beneficial for organ function recovery from acute injury. However, compensatory proliferation is not always advantageous, as the proliferating cells are vulnerable to ongoing detrimental stimuli, such as inflammation, endocrine stress, high blood pressure, hypoxia/ischemia, and the like. Cell injury and death promotes secretion of growth factors, which evokes proliferation of RTECs; entering the cell cycle makes the RTECs more vulnerable to injury and death. Under persistent stress, death and proliferation are mutually promoted and form the vicious circle that triggers, maintains, and augments the inflammation and progression of TF. We hypothesize that the "proliferation-death" circle is another important pathophysiologic mechanism of TF onset. Through this hypothesis, this paper interprets the development and progression of TF. Moreover, the vicious circle may be universal, underlying the development of inflammation and fibrosis in various organs and tissues. The hypothesis also suggests a potential therapy strategy for the inhibition of fibrosis.

Estrogen receptor alpha gene polymorphisms and breast cancer risk: a case-control study with meta-analysis combined.

Molecular epidemiological studies have shown that gene polymorphisms of estrogen receptor alpha gene (ESR-α) are associated with breast cancer risk. However, previous results from many molecular studies have been inconsistent. In this study, we examined two polymorphisms (PvuII and XbaI RFLPs) of the ESR-α gene in 542 breast cancer cases and 1,016 controls from China. Associations between the polymorphisms and breast cancer risk were calculated with an unconditional logistic regression model. Linkage disequilibrium and haplotypes were analyzed with the SHEsis software. In addition, we also performed a systematic meta-analysis of 24 published studies evaluating the association. No significant associations were found between the PvuII polymorphism and breast cancer risk. However, a significantly decreased risk of breast cancer was observed among carriers of the XbaI 'G' allele (age-adjusted OR = 0.80; 95% CI = 0.66- 0.97) compared with carriers of the 'A' allele. Haplotype analysis showed significantly decreased cancer risk for carriers of the 'CG' haplotype (OR = 0.79; 95% CI = 0.66- 0.96). In the systematic meta-analysis, the XbaI 'G' allele was associated with an overall significantly decreased risk of breast cancer (OR = 0.90, 95% CI = 0.82- 1.00). In addition, the PvuII 'C' allele showed a 0.96- fold decreased disease risk (95% CI = 0.92- 0.99). In subgroup analysis, an association between the PvuII 'C' and XbaI 'G' alleles and breast cancer risk was significant in Asians ('C' vs. 'T': OR = 0.93, 95% CI = 0.85- 1.00; 'G' vs. 'A': OR = 0.82, 95% CI = 0.68- 0.98), but not in Euro-Americans. Thus, our results provide evidence that ESR-α polymorphisms are associated with susceptibility to breast cancer. These associations may largely depend on population characteristics and geographic location.

[Establishment of a real-time PCR assay for simultaneously detecting human BKV and CMV DNA and its application in renal transplantation recipients].

To establish a fluorescent quantitative PCR method (FQ-PCR) with TaqMan probe for simultaneous detection of polyomavirus (BKV) and cytomegalovirus (CMV) and to evaluate its clinical application in the renal transplantation recipients. The conservative sequences of BKV and CMV were targeted and amplified by nested PCR technique. The PCR products were cloned into the plasmids pcDNA3. 1(+). The recombinant plasmid containing target sequences of BKV and CMV were constructed as external standards. The TaqMan-based assay was optimized. For evaluating the assay, the sensitivity was determinated by diluted standard (5 X 103-10icopies/mL), and the specificity was verified by negative control and positive control, and the precision was assessed by intra-assay coefficient of variation (ICV) through detecting standard repeatedly (20 times). A total of 480 blood samples of renal transplantation recipients were used to detect BKV and CMV DNA simultaneously with FQ-PCR, and the concentrations of FK506 were measured by ELISA. The association of DNA copy and concentrations of FK506 was analyzed. The cloned target BKV and CMV DNA was confirmed by sequencing and analysis. The sensitivity of the FQ-PCR assay reached 5 X 103 copies/ml in detecting BKV or CMV DNA. Control DNA verified the assay specifically detecting target DNA. The precision of the assay to quantif target DNA copies was acceptable (Intra-assay CV was 3.44% for BKV and 2.23% for CMV; Inter-assay CV was 4. 98% for BKV and 3.76% for CMV;). Of 480 samples, 130 samples (27. 08%) were CMV DNA positive, significantly higher than the BKV DNA positive (13.33%, 64/480, P<0.05). The positive BKV or CMV DNA was found to be associated with high concentrations of FK506 (P<0. 05). In conclusion, the developed real-time PCR assay for detecting both CMV and BKV DNA simultaneously was s high sensitive, precise and time-effectiveand could be applied in the monitoring of the CMV and BKV infection in the renal transplantation recipients.

[Expression of miRNA-29b and its clinical significances in primary hepatic carcinoma].

OBJECTIVE: To explore the clinical value of miRNA-29b expression and the combined detection of serum miRNA-29b and alpha-fetoprotein (AFP) in the diagnosis of primary hepatic carcinoma(PHC). METHODS: From January 2007 to May 2010, the serum levels of miRNA-29b and AFP from 96 healthy controls and 87 PHC patients were measured by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA) respectively. The relationship of miRNA-29b and various clinical parameters was analyzed. RESULTS: Serum levels of miRNA-29b in PHC pre-operative group (0.250 (0.124 - 0.381)) significantly decreased versus the control group [0.860 (0.587 - 1.338)] and the post-operative group (0.890 (0.637 - 1.414)) (P < 0.001). Also, the levels of AFP in PHC pre-operative group (65.4 (20.1 - 212.3)) was obviously higher than that in the control group (13.3 (7.1 - 19.8)) and the post-operative group (23.2 (11.6 - 55.7)) (P < 0.001). A lower expression of miRNA-29b was correlated with lower differentiation and higher TNM stages (P < 0.045, P < 0.001). Kaplan-Meier curve analysis revealed that PHC patients with a low serum expression of miRNA-29b had a significantly shortened overall survival when compared with a high serum expression of miRNA-29b (25.52 vs 36.94 months, P = 0.008). Multivariable Cox regression analysis indicated that the serum expression of miRNA-29b was an independent risk factor for overall survival. Relative risk was 0.482 (95% confidence interval: 0.236 - 0.985). The critical values for miRNA-29b and AFP were determined at 0.38 and 23.1 µg/L through the ROC curves. Under the critical value, the sensitivity of miRNA-29b and AFP were 75.9% and 70.1% and the specificity of miRNA-29b and AFP 89.5% and 92.7% respectively. Combined detection could increase the sensitivity up to 87.3%, and achieve a specificity of 88.5%. CONCLUSION: The combined detection of miRNA-29b and AFP aids the diagnosis of PHC and the prediction of its prognosis.

Vitamin D receptor gene polymorphisms and colorectal cancer risk: a systematic meta-analysis.

AIM: To investigate the relationship between polymorphisms present in the vitamin D receptor (VDR) gene and colorectal cancer risk, a systematic meta-analysis of population-based studies was performed. METHODS: A total of 38 relevant reports published between January 1990 and August 2010 were identified, of which only 23 qualified for this meta-analysis based on our selection criteria. Five polymorphic variants of the VDR gene, including Cdx-2 (intron 1e) and FokI (exon 2) present in the 5' region of the gene, and BsmI (intron 8), ApaI (intron 8), and TaqI (exon 9) sites present in the 3' untranslated region (UTR), were evaluated for possible associations with colorectal cancer risk. Review manager 4.2 was used to perform statistical analyses. RESULTS: In the meta-analysis performed, only the BsmI polymorphism was found to be associated with colorectal cancer risk. In particular, the BsmI B genotype was found to be related to an overall decrease in the risk for colorectal cancer [BB vs bb: odds ratio (OR) = 0.87, 95% CI: 0.80-0.94, P = 3 × 10(-4); BB vs Bb + bb: OR = 0.90, 95% CI: 0.84-0.97, P = 5 × 10(-4)]. Moreover, in subgroup analyses, the BsmI B genotype was significantly associated with colon cancer, and not rectal cancer. An absence of between-study heterogeneity was also observed. CONCLUSION: A meta-analysis of 23 published studies identified the BsmI polymorphism of the VDR gene to be associated with an increased risk of colon cancer.