Increased TT-TG distance caused by excessive tibiofemoral rotation predicts poor clinical outcomes after tibial tubercle osteotomy in recurrent patellar dislocation.

PURPOSE: To compare clinical outcome between recurrent patellar dislocation (RPD) with or without actual tibial tubercle lateralisation (TTL) after medial patellofemoral ligament reconstruction (MPFL-R) combined with tibial tubercle transfer. METHODS: From 2015 to 2018, a total of 172 knees with RPD and a tibial tubercle-trochlear groove (TT-TG) distance of > 20 mm were treated with MPFL-R combined with tibial tubercle transfer. Patients were divided into the lateralisation group (TT-PCL > 24 mm, n = 74) and the nonlateralisation group (TT-PCL ≤ 24 mm, n = 60) based on the presence or absence of actual TTL (TT-PCL > 24 mm). Clinical outcomes were assessed postoperatively at a minimum of 2 years. Second-look arthroscopic evaluations were available for 84 knees to assess cartilage damage. RESULTS: A total of 134 knees with a median follow-up time of 32 months were included. Tibiofemoral rotation (TFR) was significantly higher in the nonlateralisation group than in the lateralisation group (15.4° vs. 9.4°, P < 0.001). At the final follow-up, the nonlateralisation group had significantly lower Kujala (78.2 vs. 86.4, P = 0.001) and Lysholm (80.3 vs. 88.2, P = 0.003) scores than the lateralisation group. At the time of the second-look arthroscopic assessment, 38.9% of the patients in the nonlateralisation group showed cartilage worsening in the medial patellar facet that was significantly higher than that in the lateralisation group (38.9% vs. 12.5%, P = 0.015). CONCLUSION: Patients with RPD and an increased TT-TG distance of > 20 mm but without actual tibial tubercle lateralisation benefit less from tibial tubercle transfer than patients with actual tibial tubercle lateralisation, which may be related to the significantly higher tibiofemoral rotation angle of the former. LEVEL OF EVIDENCE: III.

Clinical and Second-look Arthroscopic Results for Derotational Distal Femoral Osteotomy With Medial Patellofemoral Ligament Reconstruction for Recurrent Patellar Dislocation With Increased Femoral Anteversion: A Series of 102 Cases With a Minimum Clinical Follow-up of 2 Years.

BACKGROUND: Derotational distal femoral osteotomy (DDFO) has been used to treat patients with recurrent patellar dislocation (RPD) with increased femoral anteversion. However, no study has reported second-look arthroscopic findings in the patellofemoral joint after DDFO. PURPOSE: To report clinical and second-look arthroscopic outcomes for DDFO with combined medial patellofemoral ligament reconstruction (MPFL-R) in treating RPD with increased femoral anteversion. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: From 2015 to 2019, 131 consecutive patients (144 knees) with RPD were treated with combined MPFL-R and DDFO. Patients with a femoral anteversion angle >30° and a minimum 2-year clinical follow-up period were included in the study. Three-dimensional computed tomography was performed to evaluate rotational deformities of the lower leg. Radiographic parameters presenting bony abnormalities associated with RPD were measured. Second-look arthroscopic evaluations were available for 86 knees to assess patellar tracking and chondral lesion changes. Moreover, clinical and radiologic outcomes were assessed pre- and postoperatively at a minimum 2 years. RESULTS: A total of 102 knees in 92 patients were included in the present study with a mean clinical follow-up of 4.1 years (range, 2.0-5.6 years). Mean ± SD femoral anteversion changed significantly from 34.7°± 7.5° preoperatively to 11.3°± 0.2° postoperatively (P < .001), and mean tibial tubercle-trochlear groove distance decreased significantly from 19.6 ± 3.5 mm preoperatively to 17.4 ± 3.2 mm postoperatively (P < .001). In the majority of knees, at the time of second-look arthroscopic assessment, chondral lesion status remained unchanged at the lateral patellar facet (96%) and trochlear groove (95%); in contrast, chondral damage at the medial patellar facet was aggravated in 9 cases (10%). All functional scores (Tegner, Lysholm, visual analog scale, and Kujala scores) improved significantly at final follow-up. None of the patients experienced redislocation or subluxation after surgery. CONCLUSION: Chondral lesions in the patellofemoral joint remained unchanged in the majority of cases in second-look arthroscopy after combined MPFL-R and DDFO. Moreover, high-grade trochlear dysplasia and arthroscopic residual patellar maltracking might be associated with cartilaginous deterioration at the medial patellar facet after surgery.

Antitumor activity of RUNX3: Upregulation of E-cadherin and downregulation of the epithelial-mesenchymal transition in clear-cell renal cell carcinoma.

RUNX3 is a transcription factor and tumor suppressor that is silenced or inactivated in diverse tumors. The effect of RUNX3 on the epithelial-mesenchymal transition in clear-cell renal cell carcinoma (CCRCC) remains unclear. We determined the expression of RUNX3 and E-cadherin in tumor tissues and adjacent normal tissues of 30 CCRCC patients; established cultured CCRCC cells with the overexpression of RUNX3; and examined the in vivo tumorigenic function of RUNX3 in a nude mouse xenograft model of CCRCC. RUNX3 and E-cadherin were downregulated in human CCRCC samples. Cell lines with RUNX3 overexpression had reduced cell proliferation, invasion, and migration, a prolonged cell cycle, increased apoptosis, and increased expression of E-cadherin. In the nude mouse xenograft model of CCRCC, tumors with the overexpression of RUNX3 had smaller volumes and weights and had increased expression of E-cadherin. In conclusion, RUNX3 overexpression increased the level of E-cadherin and inhibited the proliferation, invasion, and migration of CCRCC in vitro and in vivo. RUNX3 has potential use as a biomarker for prognostic monitoring of CCRCC and as a therapeutic target for the treatment of this cancer.

Predictors of Graft Failure After Primary Medial Patellofemoral Ligament Reconstruction.

Background: The tibiofemoral rotation angle has been found to be higher in patients with recurrent patellar dislocations (RPDs) than in healthy people; however, little is known about the clinical significance of this finding. Purpose: To determine whether an increased tibiofemoral rotation angle is associated with graft failure after primary medial patellofemoral ligament reconstruction (MPFL-R) and to investigate the role of the tibiofemoral rotation angle in predicting MPFL-R failure in patients with RPDs. Study Design: Case-control study; Level of evidence, 3. Methods: We retrospectively analyzed the records of 632 consecutive patients with clinically diagnosed RPDs from 2011 to 2018. Postoperative stress radiography of the patellofemoral joint was performed to identify whether the graft failed. After a review, 33 patients who showed MPFL-R failure were allocated to the failure group. They were matched 1:2 to 66 participants who underwent successful MPFL-R (control group). The cutoff value and area under the curve (AUC) of the tibiofemoral rotation angle for predicting graft failure after primary MPFL-R were determined, and the risk factors for MPFL-R failure were assessed by multivariate logistic regression analysis. Results: The tibiofemoral rotation angle was significantly higher in the failure group than in the control group (16.4° ± 5.6° vs 6.4° ± 4.5°, respectively; P < .001). The cutoff value of the tibiofemoral rotation angle for predicting graft failure was 12.3° (sensitivity, 81.8%; specificity, 89.4%; AUC, 0.920). Overall, 3 risk factors for MPFL-R failure were determined: excessive tibiofemoral rotation (≥12.3°) (odds ratio [OR], 13.159 [95% CI, 2.469-70.139]; P = .003), a preoperative high-grade J-sign (OR, 7.674 [95% CI, 1.232-47.809]; P = .029), and a femoral tunnel malposition (OR, 6.976 [95% CI, 1.077-45.187]; P = .042). Conclusion: In this study, excessive tibiofemoral rotation, a preoperative high-grade J-sign, and a femoral tunnel malposition were identified as risk factors for graft failure after primary MPFL-R in patients with RPDs. More importantly, excessive tibiofemoral rotation ( ≥ 12.3°) may predict the failure of primary MPFL-R, which can help surgeons easily identify high-risk patients of MPFL-R failure before surgery.

Posterior tibial slope measurements based on the full-length tibial anatomic axis are significantly increased compared to those based on the half-length tibial anatomic axis.

PURPOSE: This study aimed to compare the difference in posterior tibial slope (PTS) measurements based on the full-length and half-length tibial anatomic axes of the same group of patients. It was hypothesized that the obtained PTS values would be affected by the length of tibia chosen during the measurements. METHODS: Full-length true lateral tibia radiographs were obtained for each patient who underwent anterior cruciate ligament reconstruction (ACLR) in our department. PTS measurements were obtained by measuring the angle between the full-length or half-length tibial anatomic axis and an average of the lateral and medial tibial plateau. The anatomic axis was defined as the center of the tibial diaphysis. The PTS measurements from the full-length and half-length true lateral tibia radiographs were obtained and compared. Additionally, the absolute difference and the relationship between the two PTS measurements were calculated and analyzed. RESULTS: A total of 200 ACL-injured patients were included in this study. The average PTS values using the anatomic axis were 15.9 ± 3.7° and 14.1 ± 3.7° on full-length and half-length true lateral tibial radiographs. There was a significant difference between the measurements with the full-length and half-length tibial radiographs (P < 0.01). Additionally, 49.5% (n = 99) of patients had ≥ 2.0° differences between the full-length and half-length anatomic axis PTS measurement techniques; meanwhile, a strong and significant linear relationship (r = 0.95; P < 0.001) was identified between the two PTS measurements. CONCLUSION: There were significant differences and linear relationships between PTS measurements that measured the anatomic axis from full-length and half-length true lateral tibia radiographs. Therefore, the obtained PTS values were strongly associated with the length of tibia chosen during the measurements. Surgeons should pay more attention to the measurement techniques and the tibial length when considering the role of PTS in ACL injury and ACLR failure. Knowledge of the association is very important for calculating potential closing wedge proximal tibial osteotomies to correct excessive PTS in the setting of ACLR failures. LEVEL OF EVIDENCE: IV.

Steep Posterior Tibial Slope and Excessive Anterior Tibial Translation Are Predictive Risk Factors of Primary Anterior Cruciate Ligament Reconstruction Failure: A Case-Control Study With Prospectively Collected Data.

BACKGROUND: Steep posterior tibial slope (PTS) and excessive anterior tibial translation (ATT) have been identified as important anatomic risk factors for anterior cruciate ligament (ACL) injury, which have raised concerns about clinical outcomes after primary ACL reconstruction (ACLR). PURPOSE: To investigate anatomic risk factors of primary ACLR failure and to determine the cutoff values of PTS and ATT for predicting primary ACLR failure. STUDY DESIGN: Case-control study; Level of evidence, 3. METHODS: Between November 2015 and May 2017, a total of 215 consecutive patients with clinically diagnosed noncontact ACL injuries who underwent primary anatomic ACLR were retrospectively analyzed. Among them, 25 patients who showed complete discontinuity of ACL fibers on final follow-up magnetic resonance imaging scans were allocated into the failure group (study group). They were matched 1:2 to 50 control participants who showed clear and continuous ACL fibers on magnetic resonance imaging scans (control group). PTS and ATT were measured on preoperative weightbearing whole leg lateral radiographs and compared between the groups. The cutoff values of PTS and ATT for predicting primary ACLR failure were determined by the receiver operating characteristic curve. Moreover, predictors of primary ACLR failure were assessed by multivariate logistic regression analysis, including sex, age, body mass index, concomitant meniscal tears, degree of pivot-shift test, and KT-1000 arthrometer side-to-side difference, PTS, and ATT. RESULTS: PTS and ATT values in the study group were significantly higher than those in the control group (mean ± SD: PTS, 17.2°± 2.2° vs 14.4°± 2.8°; ATT, 8.3 ± 3.4 mm vs 4.1 ± 3.1 mm; P < .001). The cutoff values of PTS and ATT for predicting primary ACLR failure were 17° (sensitivity, 66.7%; specificity, 90.9%) and 6 mm (sensitivity, 87.5%; specificity, 79.5%), respectively. Additionally, PTS ≥17° (odds ratio, 15.6; 95% CI, 2.7-91.5; P = .002) and ATT ≥6 mm (odds ratio, 9.9; 95% CI, 1.9-51.4; P = .006) were determined to be risk factors of primary ACLR failure, whereas sex, age, body mass index, concomitant meniscal tears, degree of the pivot-shift test, and KT-1000 arthrometer side-to-side difference were not. CONCLUSION: In this study, PTS ≥17° and ATT ≥6 mm, as measured on weightbearing whole leg radiographs, were identified to be predictive risk factors of primary ACLR failure. This study adds to the existing knowledge about potential surgical indications of simultaneous slope-reducing high tibial osteotomy to mitigate the primary ACLR failure rate.

Efficacy of anti-T-lymphocyte globulin-Fresenius as an induction agent in deceased-donor renal transplantation: A cohort study.

Anti-T-lymphocyte globulin (ATG) is frequently used in the induction regimen of renal transplantation, but its dose has not been standardized. In the present study, the efficacy of different ATG-Fresenius (ATG-F) doses was assessed in recipients of renal transplantation. A total of 131 adult recipients of renal transplantation who received ATG-F induction between August 2015 and July 2018 were included. The incidence of biopsy-confirmed acute rejection, graft function, as well as graft and patient survival within 12 months post-transplant, was assessed, and adverse events, including hematologic and infection-associated side effects, were compared between patients receiving a cumulative ATG-F dose of <7 or ≥7 mg/kg. The incidence of biopsy-confirmed acute rejection was similar between patients receiving cumulative doses of <7 and ≥7 mg/kg (7.5 vs. 4.7%, P=0.766). The incidence of infection within 12 months was lower in the ATG-F <7 mg/kg group compared with that in the ≥7 mg/kg group (26.9 vs. 50.0%, P=0.006), but the incidence of pneumonia did not differ between the ATG-F <7 and ≥7 mg/kg groups (10.4 vs. 20.3%, P=0.117). The incidence of urinary infection was higher in the ≥7 mg/kg group than in the <7 mg/kg group (20.4 vs. 7.46%, P=0.033), while the extent and duration of anemia and lymphopenia was similar between groups. There was no difference in graft function, delayed graft function, as well as overall and graft survival between the groups. In conclusion, a moderate reduction in the cumulative ATG-F dose was not associated with an increased risk of acute rejection, while the risk of infection was reduced. Optimization of the ATG-F dose for induction may facilitate the reduction of the risk of infection without compromising the induction efficacy in renal transplant recipients.

Total Knee Arthroplasty Using a Medial Pivot or Posterior Cruciate-Stabilizing Prosthesis in Chinese Patients.

There is an unmet need for a prosthesis designed according to the anatomical parameters of the Chinese population. This study aims to compare the use of a medial pivot (MP) implant or posterior cruciate ligament (PCL) substitution (posterior-stabilized [PS]) prosthesis for unilateral total knee arthroplasty (TKA) in a Chinese population. The medical records of patients undergoing unilateral TKA with an MP implant (Group A) or a PS prosthesis (Group B) at our institution between January 2010 and December 2011 were retrospectively reviewed. Patients were followed up for 5 years. Preoperatively and at the December 2016 postoperative follow-up, the Hospital for Special Surgery scoring system (HSS knee score) and the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score were measured to evaluate TKA outcomes. This study included 49 patients in Group A and 51 in Group B. As of December 2016, there were no significant differences in the preoperative/postoperative changes in any category of the HSS knee score or WOMAC score between the groups. There were no postoperative complications in either group during the 5-year follow-up. There were no periprosthetic infections or need for revision surgery. One patient in Group A experienced aching and a small amount of effusion in the articular cavity that was attributed to overexertion. In conclusion, there were no significant differences in midterm outcomes in Chinese patients receiving an MP implant or a PS prosthesis for unilateral TKA. These data suggest that the MP and PCL substitution design are safe and effective for unilateral TKA in China.

[Preoperative Lymphocyte-to-monocyte Ratio Predicts Prognosis in Patients with Stage T1 Non-muscle Invasive Bladder Cancer].

Objective To investigate the clinical value of preoperative lymphocyte-to-monocyte ratio(LMR)in evaluating the prognosis of patients with stage T1 non-muscle invasive bladder cancer(NMIBC).Methods A total of 215 patients with stage T1 NMIBC who underwent transurethral resection of bladder tumor were enrolled.Clinical data were collected.Patients were followed up and their disease-free survival(DFS)and overall survival(OS)were recorded.The receiver operating characteristic(ROC)curve of preoperative LMR in detecting patient prognosis was used to determine the optimal cut-off value for LMR.Patients were divided into low LMR group(LMR <3.86,n=77)and high LMR group(LMR ≥ 3.86,n=138).Kaplan-Meier survival curves were explored to compare cumulative DFS and OS rates in patients with different LMR levels,and COX proportional hazards regression model was used to analyze factors associated with DFS and OS.Results All these 215 patients with T1 stage NMIBC were followed up for 2-92 months,and the DFS rate was 59.07% and OS rate was 65.12%.Kaplan-Meier curves showed that the cumulative DFS rate(χ 2=4.784,P=0.029)and cumulative OS rate(χ 2=7.146, P=0.008)in the low LMR group were significantly lower than those in the high LMR group.Tumor size ≥ 3 cm(HR=1.398,95% CI:1.042-1.875,P=0.025),pathological grade G3(HR=1.266,95% CI:1.026-1.563,P=0.028),and LMR ≥ 3.86(HR=2.347,95% CI:1.080-5.101,P=0.031)were independent factors associated with DFS in patients with stage T1 NMIBC.In addition,tumor size ≥ 3 cm(HR=1.228,95% CI:1.015-1.484,P=0.034),pathological grade G3(HR=1.366,95% CI:1.017-1.834,P=0.038),and LMR<3.86(HR=2.008,95% CI:1.052-3.832,P=0.035)were independent factors associated with OS in patients with T1 stage NMIBC. Conclusion Preoperative LMR is an independent factor associated with patients' prognosis in T1 stage NIMBC.Patients with low LMR tend to have higher risk of NMIBC progression and death.

Monocarboxylate transporter 1 and monocarboxylate transporter 4 in cancer-endothelial co-culturing microenvironments promote proliferation, migration, and invasion of renal cancer cells.

BACKGROUND: The Warburg effect demonstrates the importance of glycolysis in the development of primary and metastatic cancers. We aimed to explore the role of monocarboxylate transporter 1 (MCT1) and MCT4, two essential transporters of lactate, in renal cancer progression during cancer-endothelial cell co-culturing. METHODS: Renal cancer cells (786-O) and human vascular endothelial cells (HUVECs) were single-cultured or co-cultured in transwell membranes in the presence or absence of a MCT-1/MCT-4 specific blocker, 7ACC1. Cell proliferation was evaluated with the CCK-8 kit, while cell migration, after a scratch and invasion in transwell chambers, was evaluated under a microscope. Real-time qPCR and western blot were employed to determine the mRNA and protein levels of MCT1 and MCT4, respectively. The concentration of lactic acid in the culture medium was quantified with an l-Lactic Acid Assay Kit. RESULTS: 786-O cells and HUVECs in the co-culturing mode exhibited significantly enhanced proliferation and migration ability, compared with the cells in the single-culturing mode. The expression of MCT1 and MCT4 was increased in both 786-O cells and HUVECs in the co-culturing mode. Co-culturing promoted the invasive ability of 786-O cells, and markedly increased extracellular lactate. Treatments with 7ACC1 attenuated cell proliferation, migration, and invasion, and down-regulated the levels of MCT1/MCT4 expression and extracellular lactate. CONCLUSIONS: The Warburg effect accompanied with high MCT1/MCT4 expression in the cancer-endothelial microenvironments contributed significantly to renal cancer progression, which sheds new light on targeting MCT1/MCT4 and glycolytic metabolism in order to effectively treat patients with renal cancers.

Monocarboxylate transporters MCT1 and MCT4 are independent prognostic biomarkers for the survival of patients with clear cell renal cell carcinoma and those receiving therapy targeting angiogenesis.

BACKGROUND: Prognostic biomarkers for patients with clear cell renal cell carcinoma (ccRCC), particularly those receiving therapy targeting angiogenesis, are not well established. In this study, we examined the correlations of monocarboxylate transporter 1 (MCT1) and MCT4, 2 critical transporters for glycolytic metabolism, with various clinicopathological parameters as well as survival of patients with ccRCC and those treated with vascular endothelial growth factor receptor (VEGFR) inhibitors. METHODS: A cohort of 150 ccRCC patients were recruited into this study. All patients underwent radical or partial nephrectomy as the first-line treatment, and 38 received targeted therapy (sorafenib or sunitinib) after the surgery. Expression levels of MCT1, MCT4, and CD34 were examined by immunohistochemistry. Correlations between MCT1 or MCT4 expression and different clinicopathological parameters or patient survival were analyzed among all as well as patients receiving targeted therapy. RESULTS: MCT1 or MCT4 expression did not significantly correlate with sex, age, tumor diameter, microvascular density, tumor staging, pathological Furmann grade, or MSKCC (P>0.05). High expression of either MCT1 or MCT4 significantly correlated with reduced overall survival (OS) and progression-free survival (PFS) among the total cohort of ccRCC patients. For patients receiving targeted therapy, high expression of either MCT1 or MCT4 significantly correlated with reduced PFS, but not OS. Both conditions were independent prognostic biomarkers for reduced PFS among all patients or those receiving targeted therapy. CONCLUSION: MCT1 and MCT4 are prognostic biomarkers for patients with ccRCC or those receiving targeted therapy. High expression of these 2 proteins predicts reduced PFS in these patients.

Targeting of MCT1 and PFKFB3 influences cell proliferation and apoptosis in bladder cancer by altering the tumor microenvironment.

Phosphofructokinase-2/fructose-2,6-bisphosphatase 3 (PFKFB3) and monocarboxylate transporter 1 (MCT1) play important roles in tumor endothelial cells (ECs) and several biological processes. The present study was conducted to study the effects of PFKFB3 and MCT1 on cell proliferation and apoptosis in the tumor microenvironment by co-culture of HUVECs and T24, a bladder cancer (BC) cell line, using a microfluidic device. Immunofluorescence assay showed that HUVEC activity was significantly enhanced under co-culture with T24 cells, according to the stronger fluorescence intensity of CD31 and CD105 than that in the signal­cultured cells. Quercetin treatment inhibited MCT1 expression but did not affect PFKFB3 expression. Knockdown of MCT1 or/and PFKFB3 increased the apoptosis rate of the HUVECs under single-culture and co-culture situations by staining with calcein and propidium iodide. Meanwhile, cell proliferation and lactic concentration were significantly decreased after the blocking of MCT1 or/and PFKFB3, as compared with that in the control group. No obvious differences in the effects on apoptosis, proliferation and lactic concentration were found between cells treated with quercetin and siMCT1. Thus, we concluded that the targeting of MCT1 and PFKFB3 regulated cell proliferation and apoptosis in BC cells by altering the tumor microenvironment, and quercetin exhibited a potential antitumor effect by targeting MCT1.

The study of energy metabolism in bladder cancer cells in co-culture conditions using a microfluidic chip.

OBJECTIVES: This study aimed to systematically analyze changes in mitochondrial-related protein expression in bladder cancer cells and tumor-associated fibroblasts and to investigate the characteristics of bladder cancer cell energy metabolism. METHODS: In this study, we utilized the following techniques to achieve the objectives: (1) a co-culture system of bladder tumor cells and fibroblasts was built using a microfluidic chip as a three-dimensional culture system; (2) the concentration of lactic acid in the medium from the different groups was determined using an automatic micro-plate reader; (3) a qualitative analysis of mitochondria-related protein expression was performed by immunofluorescent staining; and (4) a quantitative analysis of mitochondrial-associated protein expression was conducted via Western blot. SPSS software was utilized to analyze the data. RESULTS: (1) Determination of lactic acid concentration: The lactic acid concentration was determined to be highest in the experimental group, followed by the T24 cell control group and then the fibroblast control group. (2) Qualitative results: In the control group, the mitochondrial-related protein fluorescence intensity was higher in the fibroblasts compared with the cancer cells, and the fluorescence intensity of the fibroblasts was reduced compared with the experimental group. The mitochondrial-related protein fluorescence intensity of the cancer cells was higher in the experimental group compared with the control group, and the opposite results were obtained with the fibroblasts. (3) Quantitative results: The expression of mitochondria-related proteins was higher in fibroblasts compared with cancer cells in the control group, and the opposite results were obtained in the experimental group (P<0.05). The expression of mitochondria-related proteins was increased in cancer cells in the experimental group compared with the control group; the opposite results were observed for the fibroblasts (P<0.05). CONCLUSIONS: The energy metabolism of bladder tumor cells does not parallel the "Warburg effect" because even under sufficient oxygen conditions, cancer cells still undergo glycolysis. Bladder cancer cells also have an efficient oxidative phosphorylation process wherein cancer cells promote glycolysis in adjacent interstitial cells, thereby causing increased formation of nutritional precursors. These high-energy metabolites are transferred to adjacent tumor cells in a specified direction and enter the Krebs Cycle. Ultimately, oxidative phosphorylation increases, and sufficient ATP is produced.

A bladder cancer microenvironment simulation system based on a microfluidic co-culture model.

A tumor microenvironment may promote tumor metastasis and progression through the dynamic interplay between neoplastic cells and stromal cells. In this work, the most representative and significant stromal cells, fibroblasts, endothelial cells, and macrophages were used as vital component elements and combined with bladder cancer cells to construct a bladder cancer microenvironment simulation system. This is the first report to explore bladder cancer microenvironments based on 4 types of cells co-cultured simultaneously. This simulation system comprises perfusion equipment, matrigel channel units, a medium channel and four indirect contact culture chambers, allowing four types of cells to simultaneously interact through soluble biological factors and metabolites. With this system, bladder cancer cells (T24) with a tendency to form a 'reticular' structure under 3 dimensional culture conditions were observed in real time. The microenvironment characteristics of paracrine interactions and cell motility were successfully simulated in this system. The phenotype change process in stromal cells was successfully reproduced in this system by testing the macrophage effector molecule Arg-1. Arg-1 was highly expressed in the simulated tumor microenvironment group. To develop "precision medicine" in bladder cancer therapy, bladder cancer cells were treated with different clinical 'neo-adjuvant' chemotherapy schemes in this system, and their sensitivity differences were fully reflected. This work provides a preliminary foundation for neo-adjuvant chemotherapy in bladder cancer, a theoretical foundation for tumor microenvironment simulation and promotes individual therapy in bladder cancer patients.

Bladder Tumor Heterogeneity: The Impact on Clinical Treatment.

Bladder cancer relapse and treatment failure in most patients have often been attributed to chemoresistance in tumor cells and metastasis. Emerging evidence indicates that tumor heterogeneity may play an equally important role and extends to virtually all measurable properties of cancer cells. Although the idea of tumor heterogeneity is not new, little attention has been paid to applying it to understand and control bladder cancer progression. With the development of biotechnology, such as Gene sequencing, recent advances in understanding its generation model, original basis, consequent problems, and derived therapies provide great potential for tumor heterogeneity to be considered a new insight in the treatment of bladder cancers.

Warburg effect or reverse Warburg effect? A review of cancer metabolism.

Cancer is a major threat to human health. A considerable amount of research has focused on elucidating the nature of cancer from its pathogenesis to treatment and prevention. Tumor cell metabolism has been considered a hallmark of cancer. Cancer cells differ from normal cells through unlimited cell division, and show a greater need for energy for their rapid growth and duplication. Research on glycometabolism, as the key point of energy metabolism, has played a unique role. In the 1920s, Warburg found that cancer cells prefer to produce adenosine triphosphate (ATP) by glycolysis, which is a less efficient pathway compared to oxidative phosphorylation. This striking discovery, called 'the Warburg effect', has influenced and guided the study of the mechanism and treatment of tumors for generations, but its causal relationship with cancer progression is still unclear. Some studies have now shown contradicting evidence and a new hypothesis, the reverse Warburg effect, has been put forward, in which cancer cells produce most of their ATP via glycolysis, even under aerobic conditions. In this review we discuss the new points concerning the energy metabolism of a tumor, as well as the current facts and perspectives.

Heterogeneity research in muscle-invasive bladder cancer based on differential protein expression analysis.

The aim of this study was to study the expression profiles of muscle-invasive bladder cancer (MIBC) cells of different risk groups and to explore the crucial role of biological pathway change in heterogeneity of MIBC cells. Thirty individual samples (cancer and non-cancerous specimens) were obtained from patients with MIBC. Laser capture microdissection was employed to harvest the homogeneous MIBC cells and normal urothelial cells. iTRAQ and 2D-LC-MS/MS were used to quantify and identify the differently expressed proteins. Then, the significantly changed proteins were further analyzed using Arraytrack ™ software. The interested proteins were compared with the published literatures to discuss the exact functions. A total of 3,073 non-redundant proteins were identified in this research; therefore, 855/2,210/633 (fold change >1.5 relative to normal group) presented in high-/median-/low-risk groups, respectively. 617/1,620/463 proteins with SWISS-ACC number output from Arraytrack ™ software and presented in high-/median-/low-risk groups, respectively. Pathway analysis revealed that the mainly changed pathways (top-10, p < 0.05) in Genetic information processing category were similar in high- and median-risk groups, including Kyoto Encyclopedia of Genes and Genomes (KEGG) spliceosome, protein export, ribosome pathways. The mainly altered pathways in Metabolism category included glycolysis/gluconeogenesis, pentose phosphate, pyruvate metabolism pathway for high-risk group, and glutathione metabolism, citrate cycle, oxidative phosphorylation pathways for median-risk group. The major changed pathways for low-risk group included focal adhesion pathway and ECM-receptor interaction pathway. The changed biological pathways are closely related to the regulation of heterogeneity for MIBC. The KEGG pathways of Genetic information processing category and Metabolism (anaerobic or aerobic) category play a crucial role in determining the malignant phenotype of MIBC cells. The quantification analysis of proteins combining with the KEGG pathway analysis contributes to screening candidate biomarkers and guides the biological molecular therapy of MIBC.

Far from resolved: stromal cell-based iTRAQ research of muscle-invasive bladder cancer regarding heterogeneity.

The aim of the present study was to globally characterize the cancer stroma expression profile of muscle-invasive bladder cancer in different metastatic risk groups and to discuss the decisive role of biological pathway change in cancer heterogeneity. Laser capture microdissection was employed to harvest purified muscle-invasive bladder cancer stromal cells derived from 30 clinical samples deriving from 3 different metastatic risk groups. Isobaric tags for relative and absolute quantitation (iTRAQ) and two-dimensional liquid chromatography tandem mass spectrometry (2D LC-MS/MS) were used to identify the differentially expressed proteins. Subsequently, the differentially expressed proteins were further analyzed by bioinformatics tools. After completing the above tasks, the proteins of interest were further compared with the published litterature. We identified 1,049 differentially expressed proteins by paired comparison (high risk vs. median, low risk and normal groups; median risk vs. low risk and normal groups, low risk vs. normal group; a total of 6 comparisons). A total of 510,549,548 proteins as significantly altered (ratio fold-change≥1.5 or ≤0.667 between the metastatic potential risk group and the normal group) were presented in the low/median/high metastatic risk group, respectively. Pathway analysis revealed that the differentially expressed proteins were mainly located in the Kyoto Encyclopedia of Genes and Genomes pathways, including focal adhesion pathway, systemic lupus erythematosus pathway and ECM-receptor interaction pathway. In addition, several proteins such as EXOC4, MYH10 and MMP-9 may serve as candidate biomarkers of muscle-invasive bladder cancer. Our study confirmed that stromal cells, an important part of the cancer tissue, are pivotal for regulating the heterogeneity of cancer. Common changes in biological pathways determined the malignant phenotype of muscle-invasive bladder cancer, and biomarker discovery should take into account both neoplastic cells and their corresponding stromata.

Effect of TLR4 and B7-H1 on immune escape of urothelial bladder cancer and its clinical significance.

BACKGROUND/AIM: Toll-like receptor 4 (TLR4) and B7-H1, both normally expressed restricted to immune cells, are found to be aberrantly expressed in a majority of human tumors and may play important roles in regulation of tumor immunity. It has been shown that urothelial bladder cancer (UBC) patients can manifest tumoral immune escape which may be a potential critical factor in tumor pathogenesis and progression. However, so far, the mechanisms of UBC-related immune escape have not been clarified. The aim of this study was to investigate the effect of TLR4 and B7-H1 on immune escape of UBC. METHODS: Bladder cancer T24 cells were pre-incubated with LPS and co-cultured with tumor specific CTLs. CTL cytotoxicity and apoptosis rates were measured by MTT assay and flow cytometry, respectively. The effects of an ERK inhibitor on B7-H1 expression and CTL cytotoxicity against T24 cells were also evaluated. In addition, TLR4, B7-H1 and PD-1 protein expression was analyzed by immunohistochemistry in 60 UBC specimens and 10 normal urothelia. RESULTS: TLR4 activation protected T24 cells from CTL killing via B7-H1 overexpression. However PD98059, an inhibitor of ERK, enhanced CTL killing of T24 cells by reducing B7-H1 expression. TLR4 expression was generally decreased in UBC specimens, while B7-H1 and PD-1 were greatly overexpressed. Moreover, expression of both B7-H1 and PD-1 was significantly associated with UICC stage and WHO grade classification. CONCLUSIONS: TLR4 and B7-H1 may contribute to immune escape of UBC. Targeting B7-H1 or the ERK pathway may offer new immunotherapy strategies for bladder cancer.

Cancer stroma proteome expression profile of superficial bladder transitional cell carcinoma and biomarker discovery.

PURPOSE: To globally characterize the stroma expression profile of superficial bladder transitional cell carcinoma and to discuss the cancer biology as well as biomarker discovery from stromal cells. METHODS: Laser capture microdissection was used to harvest purified bladder cancer stromal cells and normal stromal cells from 4 paired samples. Next, two-dimensional liquid chromatography-tandem mass spectrometry was used to identify the proteome expression profile. The differential proteins were further analyzed using bioinformatics tools and compared with the published literature. RESULTS: A total of 606/601 proteins commonly appeared in 4 paired samples. Six hundred and thirty-seven proteins were differentially expressed between cancer and normal stroma. Among them, 321/316 proteins specific expressed in cancer/normal stromal cells. Differential proteins were compared with the entire list of the international protein index (IPI). 36/49 GO terms exhibited as enriched and 25/14 exhibited as depleted in biological process; 39/40 GO terms exhibited as enriched and 9/4 exhibited as depleted in cellular component, respectively. Significantly altered pathways between cancer/normal stroma mainly include focal adhesion, EHEC, oxidative phosphorylation, glycolysis/gluconeogenesis, TCA cycle. Finally, descriptive statistics show the data that PI ≤ 8 have normal distribution and the proteins with extremes of PI and MW have the same probability to be a biomarker. CONCLUSIONS: The present study identified the proteome expression profile of bladder cancer stromal cells and normal stromal cells. Furthermore, our results confirm proteins origin from stroma play important role in cancer biology and highlight a novel view of cancer and stress the importance of including factors related to the cancer stroma into biomarker discovery.