A long noncoding RNA, LncRNA-LOC100127888, is associated with poor prognosis in colorectal cancer patients.

BACKGROUND: Colorectal cancer (CRC) is one of the most common malignant tumors in the world. Despite great advances in medical technology, the survival rate of CRC patients is still extremely low, mainly due to recurrence and chemotherapy resistance. Therefore, it is particularly important to find valuable biomarkers to predict the prognosis of CRC. METHODS: Immunohistochemistry was performed to test the expression of LncA in a CRC tissue microarray containing 470 tumor and corresponding normal tissues. Kaplan-Meier survival curves and a Cox proportional hazard model were used to evaluate the correlation between lncRNA-LOC100127888 (LncA) expression and CRC prognosis. Cell proliferation, migration and invasion were detected by CCK-8 and Transwell assays. RESULTS: The expression of LncA was significantly upregulated in CRC cancer tissues compared with the corresponding noncancer tissues. High LncA expression in cancer tissues was associated with pathological classification, depth of invasion, lymph node metastasis, TNM stage and distant metastasis. LncA expression was an unfavorable prognostic factor for CRC patients. Furthermore, LncA combined with clinical variables exhibited synergistic potential for the prediction of CRC prognosis. Low expression of LncA in HT 29 and HCT116 cells could decrease cell proliferation, and the migration and invasion of these cells was inhibited by knockdown of LncA. CONCLUSION: LncA could be used as an effective biomarker to predict the prognosis of CRC patients. We could predict the prognosis of CRC patients more effectively by combining LncA with clinical indicators.

Use of bioinformatic database analysis and specimen verification to identify novel biomarkers predicting gastric cancer metastasis.

Background: Gastric cancer (GC) is a common gastrointestinal tumor, and its metastasis has led to a significant increase in the death rate. The mechanisms of GC metastasis remain unclear. Methods: The differentially expressed genes (DmRs) and lncRNAs (DlncRs) of GC were selected from The Cancer Genome Atlas (TCGA) database. We applied the weighted gene co-expression network analysis (WGCNA) to construct co-expression modules related with GC metastasis. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) method analyzed the functional regions and signal pathways of genes in vital modules. DmRs-DlncRs co-expression network were drawn for finding out hub nodes. Survival analyses of significant biomarkers were analyzed by Kaplan-Meier (KM) method. Finally, the expressions of selected biomarkers were validated in cell lines and caner tissues by quantitative real-time PCR (qRT-PCR), in GC tissue microarray by Fluorescence in situ hybridization (FISH). Results: 4776 DmRs and 213 DlncRs were involved the construction of WGCNA network, and MEyellow module was identified to have more significant correlation with GC metastasis. DmRs and DlncRs of MEyellow module were proved to be involved in the processes of cancer pathogenesis by GO and KEGG pathway analysis. Through the DmRs-DlncRs co-expression network, 7 DmRs and 1 DlncRs were considered as hub nodes. Besides, the high expression of TIMD4, CETP, KRT27, PTGDS, FAM30A was worse than low expression in GC patients survival, respectively; However, LRRC26 was opposite trend. FAM30A and TIMD4 were all significant biomarkers of GC survival and hub genes. Simultaneously, TIMD4, CETP, KRT27, PTGDS, FAM30A were increased in GC cell lines and tissues compared with GES-1 and normal tissues, respectively; the expression of LRRC26 was reduced in GC cell lines and tissues. Conclusion: This study identified 6 genes as new biomarkers affecting the metastasis of GC. Especially, FAM30A and TIMD4 might be an effective marker for predicting the prognosis and a potential-therapeutic target in GC.

Stachydrine hydrochloride inhibits osteoclastogenesis by regulating the NF-κB and p38 signaling pathways to alleviate postmenopausal osteoporosis.

Osteoporosis is a common skeletal disorder characterized by low bone mass, defective bone microstructure, and increased risk of fracture. It's well known that excessive activation of osteoclasts plays a vital role in the pathogenesis of osteoporosis. Thus, inhibition of osteoclast formation and function might be a proving strategy for osteoporosis. In our study, for the first time we explored the effect of Stachydrine Hydrochloride in the treatment of osteoporosis. We demonstrated that SH markedly inhibited osteoclastogenesis and osteoclast function in vitro and effectively decrease bone resorption in vivo. These finding were further supported by changes in the NF-κB and p38 signaling pathways, which are classical downstream pathways of RANKL-mediated osteoclastogensis. Collectively, these data suggest the possible future use of SH to protect against bone loss in the treatment of osteoporosis.

miR-485-3p regulated by MALAT1 inhibits osteosarcoma glycolysis and metastasis by directly suppressing c-MET and AKT3/mTOR signalling.

AIMS: Osteosarcoma (OS) is an extremely malignant bone cancer with high incidence and rapid progression. This study aims to investigate the role and underlying mechanisms of MALAT1 and miR-485-3p in OS. MATERIALS AND METHODS: qRT-PCR and Western blotting were utilized to measure the levels of miR-485-3p, MALAT1, c-MET, AKT3, p-mTOR, mTOR, glycolysis-related proteins or migration-related proteins. Colony formation and transwell assay were used to test the roles of miR-485-3p, MALAT1, c-MET and AKT3 in cancer cell proliferation, migration and invasion. Dual luciferase assay was used to validate the interactions of miR-485-3p/c-MET, miR-485-3p/AKT3, and MALAT1/miR-485-3p. Glucose uptake assay and measurement of lactate production were employed to determine the glycolysis process. Mouse tumour xenograft model was used to determine the effect of shMALAT1 and miR-485-3p mimics on tumour growth and metastasis in vivo. KEY FINDINGS: miR-485-3p was decreased while c-MET, AKT3, and MALAT1 were increased in human OS tissues and cells. miR-485-3p bound directly to c-MET and AKT3 mRNAs and repressed OS cell glycolysis, proliferation, migration, and invasion through decreasing glycolysis-related proteins and migration-related proteins via inhibiting c-MET and AKT3/mTOR pathway. In addition, MALAT1 interacted with miR-485-3p and disinhibited c-MET and AKT3/mTOR signalling. Knockdown MALAT1 or overexpression of miR-485-3p restrained OS tumour growth and lung metastasis in vivo. SIGNIFICANCE: miR-485-3p suppresses OS glycolysis, proliferation, and metastasis via inhibiting c-MET and AKT3/mTOR signalling and MALAT1 acts as a sponge of miR-485-3p. MALAT1 and miR-485-3p may be the key regulators in OS progression, and potential molecular targets for future OS therapy.

Calculation of displacements and internal forces of anchored retaining piles.

Pile-anchor retaining structures are widely used in excavation engineering. The evaluation of lateral displacements, the internal forces of piles are extremely important for the performance of the structure. Most of the existing methods are empirical, semiempirical or FEM methods, while analytic calculation methods for this evaluation are rare. This paper presents an analytic method to calculate the displacements and internal forces of anchored retaining piles based on the existing design code. In the calculation method, the singular function is applied to evaluate the effect of segmented loading on the deflection of a beam with a nonuniform cross section. The load concentration function, expressed by the singular function, can describe the segmented load and be integrated without a complicated procedure for determining the integral constants. The method is applied to a structure in Wenzhou, China, and the calculation results are compared to the field measurement results. This method is only valid for pre-failure predictions.

DJ­1 is a new prognostic marker and predicts chemotherapy efficacy in colorectal cancer.

Protein/nucleic acid deglycase DJ­1 (DJ­1) is a 20­kDa conserved protein, which belongs to the DJ­1/ThiJ/Pfp â…  protein superfamily. Immunohistochemistry was performed to investigate the expression of DJ­1 in a colorectal cancer (CRC) tissue microarray containing tumor and corresponding adjacent normal tissues. In the present study, DJ­1 expression was significantly upregulated in CRC cells and tissues, compared with that in normal colon cells and adjacent normal tissues, respectively. In addition, patients with high DJ­1 expression levels had a worse overall survival (OS) compared with patients with low expression levels. Multivariate Cox regression analysis revealed that high DJ­1 expression levels was an independent prognostic factor for patients with CRC. Moreover, DJ­1 was able to regulate the PI3K/Akt/p27/cyclin E and PI3K/Akt/mTOR signaling pathways to promote CRC cell growth and metastasis in vitro and in vivo. In addition, DJ­1 regulated the NF­κB/Snail signaling pathway to induce CRC cell epithelial­mesenchymal transition to promote migration and invasion. Notably, patients receiving LFP treatment (oxaliplatin, 5­FU and tetrahydrofolate) had an increased OS compared with patients who underwent only surgery and low DJ­1 expression levels. The findings from the present study suggest that DJ­1 may serve as a promising prognostic marker and predicts chemotherapy efficacy in patients with CRC.

COX-2 promotes metastasis and predicts prognosis in gastric cancer via regulating mTOR.

Aim: Gastric cancer (GC) is one of the most common malignant tumors in the world. It is important to find accurate and reliable biomarkers in order to decrease whole morbidity and mortality. Results: We examined the expression of COX-2 and mTOR on GC tissue microarrays by immunohistochemistry. Multivariate COX regression analysis showed that the expression of COX-2 or mTOR was an independent factor in the prognosis of GC patients. In addition, COX-2 and mTOR have a potentially synergistic effect on predicting the prognosis of GC. Conclusion: The combined expression of COX-2 and mTOR could serve as efficient prognostic indicators and COX-2 could suppress GC metastasis via regulating mTOR.

CHIP-mediated ubiquitination of Galectin-1 predicts colorectal cancer prognosis.

CHIP and Galectin-1 are associated with the development of metastasis in cancer. However, the precise roles of CHIP or Gal1 in colorectal cancer are uncertain. Here, our study explored the relationship and clinical significance of CHIP or Gal1 in CRC. CHIP or Gal1 expression was significantly decreased or up-regulated in CRC compared with adjacent noncancerous tissues by immunohistochemistry on a CRC tissue microarray, respectively. Low CHIP or high Gal1 expression significantly correlated with clinicopathological characteristics in patients, as well as with shorter overall survival. Multivariate Cox regression analysis revealed that CHIP or Gal1 expression was an independent prognostic factor for CRC patients. Moreover, CHIP associated with Gal1 has a synergistic effect on the prediction of CRC prognosis. In vitro and vivo, high CHIP or low Gal1 expression inhibit CRC growth or metastasis. Our results found that CHIP could degradate Gal1 by ubiquitination. In summary, CHIP could inhibit CRC growth or metastasis through promoting Gal1 ubiquitination and degradation by proteasome. CHIP and Gal1 expressions are novel candidate prognostic markers in CRC. A combined effect of CHIP and Gal1 as efficient prognostic indicators was found for the first time.

Expression of JWA and XRCC1 as prognostic markers for gastric cancer recurrence.

Gastric cancer is one of the common gastrointestinal tumors. Tumor recurrence leads to a high death rate of gastric cancer. It is very important to find markers to effectively predict gastric cancer recurrence. We constructed a gastric cancer tissue microarray containing 89 tumors and corresponding normal tissues to explore the relationship between some proteins' expression and gastric cancer recurrence. The expression of JWA, Cullin1, p53, XRCC1, CHIP, FAK, MMP-2, MDM2 and p21 was determined on the microarray by immunohistochemistry. The relationship between the expression of these proteins and gastric cancer recurrence was analyzed. Tumor diameter, lymph node metastasis, and TNM stage were closely related with gastric cancer recurrence by Fisher's exact test (P<0.05). We used the univariate Cox regression analysis to find that JWA, XRCC1 were related to gastric cancer recurrence (P<0.05); Lymph node metastasis and TNM stage were closely related to gastric cancer recurrence (P<0.05). Multivariate Cox regression analysis revealed that XRCC1 or lymph node metastasis were independent risk factors of gastric cancer recurrence (P<0.05). Kaplan-Meier survival curve assay indicated that patients with low JWA or XRCC1 expression in gastric cancer had significantly shorter DFS than those with high-expressed proteins (P<0.05). JWA or XRCC1 may be effective markers to predict gastric cancer recurrence.

TIMP-2 inhibits metastasis and predicts prognosis of colorectal cancer via regulating MMP-9.

Colorectal cancer has a common cause of morbidity and mortality. Therefore, it is urgent to detect reliable biomarkers to predict prognosis in CRC. Here, we determined the expression of TIMP-2 and MMP-9 in a  CRC tissue microarray by immunohistochemistry. We found that lower TIMP-2 or/and higher MMP-9 expression in cancer tissues was correlated with poorer overall survival (OS). TIMP-2 or MMP-9 expression was independent prognostic factors for CRC. Furthermore, TIMP-2 and MMP-9 expression had a synergistic role as efficient prognostic indicators for CRC patients. In vitro and in vivo, TIMP-2 could inhibit HCT 116 cells invasion and migration by regulating MMP-9. In sum, a combined expression of TIMP-2 and MMP-9 as efficient prognostic indicators was found for the first time.

Synergistic role of Caspase-8 and Caspase-3 expressions: Prognostic and predictive biomarkers in colorectal cancer.

BACKGROUND: Expressions of Caspase-8 and Caspase-3 have been identified as important markers in many malignant tumors, but their roles in colorectal cancer (CRC) have not been confirmed. The purpose of this study was to investigate the role of Caspase-8 and Caspase-3 in CRC. METHODS: We enrolled 470 CRC patients in this study. Archival paraffin-embedded CRC tissue samples were used to construct tissue microarray (TMA), expressions of Caspase-8 and Caspase-3 that were stained by immunohistochemistry. Prognostic and predictive role of Caspase-8 and Caspase-3 expressions, alone or united, were evaluated by univariate and multivariate analysis respectively. RESULTS: In comparison with adjacent normal tissues, Caspase-8 and Caspase-3 protein levels were upregulated in CRC tissues significantly, furthermore, high expressions of Caspase-8 and Caspase-3 were correlated with decreased overall survival (OS) (p< 0.05), and also with unfavorable clinicopathologic characteristics. Cox regression analysis showed that high Caspase-8 and Caspase-3 expressions were independent negative markers of OS. CONCLUSION: Caspase-8 and Caspase-3 expressions in tumor tissues are novel candidate prognostic markers for CRC patients. It was the first time to be identified that Caspase-8 and Caspase-3 expressions had synergistic role as efficient prognostic indicators for CRC patients.

Synergistic role of Cul1 and c-Myc: Prognostic and predictive biomarkers in colorectal cancer.

Colorectal cancer (CRC) is one of the most common malignant tumors, and its high rates of recurrence and metastasis are the important causes of treatment failure in CRC. Therefore, the development of valuable molecular markers to accurately predict the prognosis of CRC patients is vital. In the present study, we determined the expression of Cullin1 (Cul1) and c-Myc in a CRC tissue microarray containing 470 cancer and corresponding normal tissues by immunohistochemistry. We found that Cul1 and c-Myc expression was significantly upregulated in the CRC cancer tissues compared with that noted in the adjacent non-cancer tissues. High Cul1 expression in cancer tissues was associated with depth of invasion (P=0.005), lymph node metastasis (P=0.001) and TNM stage (P=0.015). High c-Myc expression in cancer tissues was significantly positively association with age (P=0.004), depth of invasion (P<0.001), lymph node metastasis (P<0.001) and TNM stage (P<0.001). Multivariate Cox regression analysis revealed that Cul1 or c-Myc expression was an independent and unfavorable prognostic factor for CRC patients [hazard ratio (HR), 0.749, 95% confidence interval (CI), 0.563-0.996, P<0.05; and HR, 0.384, 95% CI, 0.257-0.472, P<0.001, respectively]. Furthermore, Cul1 and c-Myc exhibited synergistic potential for the prediction of CRC prognosis, and the patients with low expression of both Cul1 and c-Myc had a favorable survival outcome (P<0.001).

Synergistic efficacy of Cullin1 and MMP-2 expressions in diagnosis and prognosis of colorectal cancer.

BACKGROUND: Cullin1 and MMP-2 have been identified as important markers in various cancers, but their roles in colorectal cancer (CRC) have remained to be discovered. The aim of this study was to investigate the expression pattern and significance of Cullin1 and MMP-2 in CRC. METHODS: A total of 470 CRC patients were enrolled. Archival paraffin-embedded CRC tissue samples were used to generate tissue microarray blocks, which were immunohistochemically stained for Cullin1 and MMP-2. Prognostic and predictive role of Cullin1 and MMP-2 expression was evaluated by univariate and multivariate analysis, respectively. Values of p < 0.05 were considered statistically significant. RESULTS: Cullin1 and MMP-2 protein levels were significantly upregulated in CRC tissues compared with adjacent noncancerous tissues. High tumoral Cullin1 or MMP-2 expression significantly correlated with shorter overall survival (OS), as well as with clinicopathologic characteristics in patients. Multivariate regression analysis showed that high Cullin1 and MMP-2 expressions, separately and together, were independent negative markers of OS. CONCLUSION: Cullin1 and MMP-2 expressions could be novel diagnostic and prognostic markers for CRC patients.

Two-stage total hip arthroplasty for patients with advanced active tuberculosis of the hip.

BACKGROUND: Treatment of advanced active tuberculosis (TB) of the hip is confronted with great challenges. Although one-stage total hip arthroplasty (THA) is considered as a safe procedure for most patients by some authors, there are still exceptions. The purpose of this paper was to investigate the feasibility and effectiveness of two-stage THA for selected patients with advanced active TB of the hip. METHODS: Nine consecutive patients with advanced active tuberculous arthritis of the hip were reviewed in this study. Out of these nine patients, the hips of five were destroyed extensively with difficulties of thorough debridement at one operation, and the hips of the other four were detected of sinus tracts. Nine patients received the two-stage total hip arthroplasty (THA) protocol and the perioperative antituberculous medication between January 2008 and December 2013. During the first stage, a debridement was carried out after at least 2 weeks of antituberculous chemotherapy to remove abscesses and infected and necrotic tissues as thoroughly as possible, followed by antituberculous chemotherapy for a minimum of 3 months (average 4.2 months). During the second stage, hip prosthesis was implanted if the erythrocyte sedimentation rate (ESR) and the C-reactive protein (CRP) were normal and the wound was well healed. Antituberculous chemotherapy was continued for 6-9 months postoperatively to constitute a total duration of a minimum of 12 months after the first operation. The patients were then evaluated based on the reactivation of infection, the Harris hip score system, X-ray, ESR, and CRP. RESULTS: The average follow-up was 40 months (range, 18-72 months). No reactivation of TB or superimposed infection was observed in all patients. The ESR and CRP returned to the normal level with no liver injury. The average Harris hip score was increased from 35 (range, 15-55) preoperatively to 91.5 (range, 83-97) at the final follow-up. The X-ray film showed no prosthesis shift or loosening. CONCLUSIONS: Two-stage THA is an alternative treatment option for patients with advanced active tuberculosis of the hip under some difficult conditions. The hip with sinus tracts or destroyed extensively with difficulties of thorough debridement at one operation may be regarded as indications.

[Arthroscopic anatomical medical patellofemoral ligament reconstruction to treat recurrent patellar dislocation].

OBJECTIVE: To investigate the procedure and efficacy of anatomical medial patellofemoral ligament (MPFL) reconstruction for the treatment of recurrent patellar dislocation assisted with arthroscopy.
 METHODS: Between January, 2010 and December 2012, 13 patients with recurrent patellar dislocation, who underwent anatomical MPFL reconstruction and the grafts of operation, were all adopted with autograft semitendinosus. The patellar side used the modified double bone tunnels and the minimally invasive percutaneous grafts through double patellar bone tunnels, and then fixed in the femoral tunnel with absorbable interference screw. Follow-up included the records of the subjective feeling, patellar apprehension test, recurrent dislocation, CT evaluation of bone tunnel position and patellar tilt angle. Knee function was evaluated by the Lysholm score and Kujala score. 
 RESULTS: Twelve patients were followed up for 36 months (range 24-60 months). All patients were satisfied with the treatment. No recurrent dislocation occurred. All the patients showed negative apprehension test. Two patients felt uncomfortable after excessive activity in the knee. Another 2 cases lost 10° flexion than the healthy knee. CT showed that the bone tunnel position were all well. The patellar tilt angle was decreased from 20.52°±1.48° preoperative to 13.52°±1.32° postoperative, with significant difference (t=14.88, P<0. 05); the Kujala score was improved from 55.2±4.51 preoperative to 93.8±3.87 postoperative, with significant difference (t=-36.238, P<0.05); and the Lysholm score was improved from 56.68±5.52 to 93.08±4.68, with significant difference (t=-33.382, P<0.05). 
 CONCLUSION: MPFL reconstruction assisted with arthroscopy is an effective surgical procedure for the treatment of recurrent patellar dislocation, which can improve the knee function with little trauma and complications.

[Co-inhibition effect of 17-DMAG and oxaliplatin on proliferation and invasion of colorectal cancer cells].

OBJECTIVE: To explore the effect of heat shock protein 90 (HSP90) inhibitor (17-DMAG) and oxaliplatin on the proliferation and invasion of colorectal cancer. METHODS: After 17-DMAG, oxaliplatin and half-dose combination of 2 drugs processing colorectal cancer SW480 and HCT116 cell lines, CCK8 assay was applied to detect cell viability. RT-PCR and Western blot were used to detect the expression level of the apoptosis-related molecules. Transwell chemokine axis experiment and Western blot were employed to detect cell invasion ability and the expression level of tumor metastasis-associated protein. RESULTS: The growth of SW480 and HCT116 cells was inhibited after the administration of 17-DMAG and oxaliplatin(P<0.05) in dose- and time-dependent manner. Processed by 17-DMAG 100 nmol/L, oxaliplatin 50 mg/L and half-dose combination of 2 drugs, transcription level of the apoptosis inhibitory gene (Bcl-2) in SW480 and HCT116 cells was decreased, the level of apoptosis promoting gene (Bax) transcription and protein PARP-1 spliceosome expression was increased, and the above trend was more obvious when using half-dose combination of 2 drugs. Transwell chemokine axis experiments showed the penetrating relative percentage and expression level of MMP9 and integrin ß3 decreased, especially for half-dose combination of 2 drugs. CONCLUSION: 17-DMAG and oxaliplatin can co-inhibit the proliferation and invasion of colorectal cancer.

Cullin1 is a novel prognostic marker and regulates the cell proliferation and metastasis in colorectal cancer.

PURPOSE: To investigate the precise function of Cullin1 (CUL1) in colorectal cancer (CRC). METHODS: Immunohistochemistry was performed to test the expression of CUL1 on a CRC tissue microarray containing the tumor and corresponding normal tissues. Simultaneously, the correlation of CUL1 expression with clinicopathological parameters and survival was evaluated. CUL1 was over-expressed or knocked down in HCT116 and SW480 cells, then the cell proliferation, migration and invasion assays in vitro and in vivo were performed. RESULTS: In this study, we found that CUL1 expression was significantly up-regulated in CRC compared with normal colon tissues. High CUL1 expression was positively associated with lymph node metastasis (P = 0.007) and tumor diameter (P = 0.052). Multivariate Cox regression analysis revealed that high CUL1 expression was an independent unfavorable prognostic factor for CRC patients (HR = 13.9, 95% confidence interval = 5.89-32.6, P < 0.001). Moreover, we found that CUL1 over-expression induced CRC cell proliferation and the growth of xenografts in nude mice via the changing of cell-cycle proteins. In addition, increased CUL1 expression in CRC cells significantly promoted cell migration and invasion abilities in vitro and peritoneal metastasis in vivo through inducing high expression of MMPs. CONCLUSION: Our findings imply that CUL1 may serve as promising prognostic markers in CRC patients.

The prognostic value of CD133 expression in non-small cell lung cancer: a meta-analysis.

CD133 has been identified as a potential cancer stem cell (CSC) marker in non-small cell lung cancer (NSCLC). However, the clinical and prognostic significance of CD133 in NSCLC remains controversial. In this study, a meta-analysis with a total number of 13 studies was performed to clarify the association between CD133 expression and clinical outcomes in publications up to June 2013. Odds ratios (ORs) and their 95 % confidence intervals (CIs) were used to assess the association between CD133 expression and the clinicopathological characteristics of NSCLC. Hazard ratios (HRs) and their 95 % CI were used to quantify the predictive ability of CD133 on NSCLC prognosis. Analysis of these data showed that CD133 expression was not associated with any clinicopathological parameters except for histology (pooled OR = 1.35, 95%CI = 1.04-1.76, P = 0.024) and tumor differentiation (pooled OR = 3.19, 95%CI = 1.10-9.21, P = 0.032). Simultaneously, we also found that positive CD133 expression was not associated with disease-free survival (DFS) (pooled HR = 1.76, 95 % CI = 0.87-3.57, P = 0.114, random-effect) but was associated with overall survival (OS) (pooled HR = 2.06, 95 % CI = 1.08-3.91, P = 0.027, random-effect). Overall, it is appropriate to regard CD133 expression as a potential prognostic factor for the OS of NSCLC patients.

Expression of PPARgamma and Cbfalpha1 mRNA in bone marrow cells in Type 2 diabetic rats and its correlation with impaired fracture healing.

OBJECTIVE: To investigate the relationship between impaired fracture healing and the expression of peroxisome proliferator activated-receptor gamma (PPARgamma) and core binding factor alpha 1 (Cbfalpha1) mRNA in the bone marrow of Type 2 diabetic rats. METHODS: Thirty male Sprague-Dawley rats were divided into a control group (n=15, normal diet) and an experiment group (n=20, high fat and sucrosum diet). After 8 weeks, one eyeball was culled for blood collection. The experiment group was intraperitoneally injected with streptozotocin and the control group received citrate buffer. After another 2 weeks, the other eyeball was culled again to collect blood. The model of distraction osteogenesis in the left tibias of the rats was established. After 14 days, we sacrificed all the rats and collected the blood and left tibias. Both femurs were harvested with germ free. We observed the callus formation in the distraction gap by X-ray and the formation of primary matrix front and microcolumn by histological examination. We observed adipocytes in adjacent bone marrow of left tibia and computed the percentage of adipocytes accounting for the area of bone cavity. We measured the expression of PPARgamma and Cbfalpha1 mRNA in femurs marrow by RT-PCR. RESULTS: In the experimental rats, the level of triglyceride was obviously elevated (P<0.01), so was the total cholesterol (P<0.05), while the level of fasting blood glucose and serum insulin did not obviously differ (P>0.05) after 8 weeks. The level of fasting blood glucose and triglyceride was obviously elevated (P<0.01), and so was the total cholesterol (P<0.05), while the level of serum insulin was not obviously different in the experiment group after 10 and 12 weeks. Callus formation in the distraction gap was obviously diminished by X-ray in the experimental rats. The array of microcolumn formation was disordered and the area of primary matrix front was catachromasis in the controls by histology examination. The number of adipocytes in both the bone marrow and the percentage of adipocyte accounting for the area of bone cavity increased (P<0.01) in the experiment group. In the experiment group, the expression of PPARgamma mRNA was up-regulated (P<0.05) while Cbfalpha1 mRNA was down-regulated (P<0.05) in double femur marrow. CONCLUSION: Increased expression of PPARgamma mRNA and decreased expression of Cbfalpha1 mRNA in the bone marrow may cause impaired fracture healing in rats with Type 2 diabetes.