Integrative Single Cell Atlas Revealed Intratumoral Heterogeneity Generation from an Adaptive Epigenetic Cell State in Human Bladder Urothelial Carcinoma.

Intratumor heterogeneity (ITH) of bladder cancer (BLCA) contributes to therapy resistance and immune evasion affecting clinical prognosis. The molecular and cellular mechanisms contributing to BLCA ITH generation remain elusive. It is found that a TM4SF1-positive cancer subpopulation (TPCS) can generate ITH in BLCA, evidenced by integrative single cell atlas analysis. Extensive profiling of the epigenome and transcriptome of all stages of BLCA revealed their evolutionary trajectories. Distinct ancestor cells gave rise to low-grade noninvasive and high-grade invasive BLCA. Epigenome reprograming led to transcriptional heterogeneity in BLCA. During early oncogenesis, epithelial-to-mesenchymal transition generated TPCS. TPCS has stem-cell-like properties and exhibited transcriptional plasticity, priming the development of transcriptionally heterogeneous descendent cell lineages. Moreover, TPCS prevalence in tumor is associated with advanced stage cancer and poor prognosis. The results of this study suggested that bladder cancer interacts with its environment by acquiring a stem cell-like epigenomic landscape, which might generate ITH without additional genetic diversification.

Whole transcriptome sequencing identifies a competitive endogenous RNA network that regulates the immunity of bladder cancer.

Several types of non-coding RNAs such as circRNAs, lncRNAs, and miRNAs have been identified to regulate mRNAs through the mechanism known as the competitive endogenous RNA (ceRNA) network. To explore the role of the ceRNA regulatory network in the immune microenvironment of bladder cancer, whole-transcriptome sequencing of bladder tumor and its peritumoral tissues from 38 bladder cancer patients, with a total of 63 samples, was performed to screen differentially expressed circ-, lnc-, mi-, and mRNAs to construct a circ/lnc-mi-mRNA regulatory network with pruning algorithms. We excavated a key immune-related gene BDNF to build the final ceRNA network as hsa-miR-107 sponged by hsa-circ-000211, AC108488.1, and LINC00163. Finally, a meta-analysis of 7 public datasets demonstrated that low expression of BDNF and high expression of hsa-miR-107 were associated with longer survival. Our study identified a ceRNA regulatory network as a potentially new prognostic marker and molecular therapeutic target of bladder cancer.

Using a two-step method of surface mechanical attrition treatment and calcium ion implantation to promote the osteogenic activity of mesenchymal stem cells as well as biomineralization on a ß-titanium surface.

Surface treatment is known as a very efficient measure by which to modulate the surface properties of biomaterials in terms of grain structure, topography, roughness and chemistry to determine the osseointegration of implants. In this work, a two-step method of surface modification was employed to impart high osteogenic activity and biomineralization capacity on a Ti-25Nb-3Mo-2Sn-3Zr alloy (a type of ß-titanium named TLM). The preliminary surface mechanical attrition treatment (SMAT) refined the average grain size from 170 ± 19 µm to 74 ± 8 nm in the TLM surface layer and promoted the surface to be much rougher and more hydrophilic. The subsequent Ca-ion implantation did not change the surface roughness and topography obviously, but enhanced the surface wettability of the SMAT-treated TLM alloy. The in vitro evaluations of the adhesion, proliferation, osteogenic genes (RUNX2, ALP, BMP-2, OPN, OCN and COL-I) and protein (ALP, OPN, OCN and COL-I) expressions, as well as extracellular matrix (ECM) mineralization of mesenchymal stem cells (MSCs) revealed that the initial SMAT-treated sample significantly enhanced the adhesion and osteogenic functions of MSCs compared to an untreated TLM sample, and the subsequent introduction of Ca ions onto the SMAT-derived nanograined sample further promotes the MSC adhesion, proliferation, osteo-differentiation and ECM mineralization due to the adsorption of more proteins such as laminin (Ln), fibronectin (Fn) and vitronectin (Vn) on the surface, as well as the increase in extracellular Ca concentrations. In addition, the biomineralization capacity of the samples was also evaluated by soaking them in simulated bodily fluid (SBF) at 37 °C for 28 days, and the results showed that the Ca-ion implanted sample significantly boosted the deposition of Ca and P containing minerals on its surface, which was associated with the generation of more Ti-OH groups on the surface after ion implantation. The combination of the SMAT technique and Ca-ion implantation thus endowed the TLM alloy with outstanding osteogenic and biomineralization properties, providing a potential means for its future use in the orthopedic field.

Single-cell sequencing and its applications in bladder cancer.

Bladder cancer is the most common malignant tumour of the urinary system that is characterised by significant intra-tumoural heterogeneity. While large-scale sequencing projects have provided a preliminary understanding of tumour heterogeneity, these findings are based on the average signals obtained from the pooled populations of diverse cells. Recent advances in single-cell sequencing (SCS) technologies have been critical in this regard, opening up new ways of understanding the nuanced tumour biology by identifying distinct cellular subpopulations, dissecting the tumour microenvironment, and characterizing cellular genomic mutations. By integrating these novel insights, SCS technologies are expected to make powerful and meaningful changes to the current diagnosis and treatment of bladder cancer through the identification and usage of novel biomarkers as well as targeted therapeutics. SCS can discriminate complex heterogeneity in a large population of tumour cells and determine the key molecular properties that influence clinical outcomes. Here, we review the advances in single-cell technologies and discuss their applications in cancer research and clinical practice, with a specific focus on bladder cancer.

STAT3 as a target for sensitizing prostate cancer cells to irradiation.

Radioresistance of prostate cancer (PCa) is a major factor leading to local failure of radiotherapy. STAT3 is an oncogenic protein that was recently found to be activated in PCa tumors. This study aimed to investigate the radiosensitization effect of targeting STAT3 in PCa tumors. Here, the radiosensitization effect of STAT3 blockade was investigated by clonogenic assay, flow cytometry and western blot analysis in human PCa cells in vitro and in vivo. We demonstrated that STAT3 blockade with a STAT3 inhibitor or siRNA increased the radiosensitivity of PCa cells and that radiation together with STAT3 blockade induced more apoptosis and double-strand breaks (DSBs) than radiation alone in LNCaP cells. In addition, radiation induced STAT3 activation and survivin expression in PCa cells, which was inhibited by STAT3 blockade. Transfection with survivin cDNA attenuated the radiosensitization effect of STAT3 blockade. These effects were further confirmed by in vivo studies, which showed that the STAT3 inhibitor enhanced the treatment efficacy of radiation on LNCaP xenografts with decreased STAT3 activation and survivin expression. These findings suggest that STAT3 blockade radiosensitizes PCa cells through regulation of survivin. Thus, our study has revealed STAT3 as a potential sensitizer for irradiation in PCa cells. Its clinical application as an adjuvant in radiotherapy of PCa should be explored in the future.

LncSOX4 serves an oncogenic role in the tumorigenesis of epithelial ovarian cancer by promoting cell proliferation and inhibiting apoptosis.

Epithelial ovarian cancer is one of the primary causes of gynecological cancer mortality. Increasing evidence has suggested that long non­coding RNAs (lncRNAs) may serve a pivotal role in cancer development. To determine whether Lnc SRY­box 4 (SOX4), an lncRNA, promotes the self­renewal of liver tumor cells and contributes to the development of epithelial ovarian cancer, the present study investigated the expression of LncSOX4 in clinical epithelial ovarian cancer tissues and non­cancer controls by reverse transcription­quantitative polymerase chain reaction analysis. In addition, siRNA targeting LncSOX4 was designed and transfected into epithelial ovarian cancer cells to further assess the effect of knocking out LncSOX4 on cellular apoptosis, cell viability, proliferation and the cell cycle. The results demonstrated that the LncSOX4 expression level was significantly upregulated in epithelial ovarian cancer tissues (3.98 vs. 1.71, P<0.001). Silencing LncSOX4 in the SKOV3 and OVCAR3 cell lines significantly impaired cell proliferation (P<0.001). Cell cycle assays revealed that the proportion of cells in the G0/G1 phase increased significantly, whereas those in the S phase and G2/M phase decreased. Apoptosis rate additionally increased following knockdown of LncSOX4 in the two cell lines. Furthermore, it was observed that an increased LncSOX4 expression level was positively associated with larger tumor sizes, more advanced tumor grade and more distant metastases.

Laparoscopic TME associated with neoadjuvant chemoradiation towards aggressive colorectal cancer.

OBJECTIVE: This study aims to investigate the clinical synchronization of the neoadjuvant chemoradiation (NC) and the laparoscopic total mesorectal excision (TME) in the treatment of locally aggressive colorectal cancer (LACC). METHODS: 92 LACC patients were selected for the research, among who 46 cases, who were performed the synchronized NC, were divided into the treatment group, after having rest for 4-6 weeks after the treatment, the 40 patients of the treatment group, who were performed the laparoscopic surgery, formed the laparoscopy group. The rest 46 patients were divided into the control group, who were performed the conventional treatment. The intraoperative conditions, postoperative recoveries, postoperative complications and recurrence rates of the two groups were compared. RESULTS: The stage-declining rate of the treatment group was 67.3%, and the surgical resection rate, anal preservation rate and postoperative complications were 86.9%, 69.6% and 26%, respectively, which were significantly higher than the control group; while the long-term recurrence rate significantly decreased to 21.7%, and the difference was statistically significant (P<0.05). CONCLUSION: The NC could effectively achieve the stage-declining purpose against the LACC, improve the resection rate and reduce the postoperative recurrence rate.

ATPase inhibitory factor 1 is a prognostic marker and contributes to proliferation and invasion of human gastric cancer cells.

ATPase inhibitory factor 1 (IF1) has recently been considered as a potential oncogene in human cancers. Recently, increasing evidences have shown that IF1 is overexpressed in human cancers and promotes tumor growth and metastasis. In this study, we found that the expression levels of IF1 protein and mRNA in gastric cancer tissues were significantly higher than those in matched adjacent nontumor tissues. Clinical association analysis revealed that the positive expression of IF1 was correlated with large tumor size, lymph node metastasis, venous infiltration and advanced TNM tumor stage in gastric cancer. Furthermore, IF1 was an independent prognostic marker for predicting 5-year overall survival and disease-free survival of gastric cancer patients. In vitro studies found that IF1 knockdown significantly inhibited cell proliferation and induced apoptosis in SGC-7901 cells. In a nude mouse xenograft model, IF1 knockdown prominently slowed down tumor growth with reduced number of Ki-67 positive cancer cells and elevated number of TUNEL positive cancer cells. Otherwise, IF1 knockdown evidently reduced SGC-7901 cell migration and invasion. In conclusion, these results indicate that IF1 may serve as a prognostic marker and promotes tumor growth and metastasis in gastric cancer.

Gastric cancer differentiation using Fourier transform near-infrared spectroscopy with unsupervised pattern recognition.

The manuscript has investigated the application of near-infrared (NIR) spectroscopy for differentiation gastric cancer. The 90 spectra from cancerous and normal tissues were collected from a total of 30 surgical specimens using Fourier transform near-infrared spectroscopy (FT-NIR) equipped with a fiber-optic probe. Major spectral differences were observed in the CH-stretching second overtone (9000-7000 cm(-1)), CH-stretching first overtone (6000-5200 cm(-1)), and CH-stretching combination (4500-4000 cm(-1)) regions. By use of unsupervised pattern recognition, such as principal component analysis (PCA) and cluster analysis (CA), all spectra were classified into cancerous and normal tissue groups with accuracy up to 81.1%. The sensitivity and specificity was 100% and 68.2%, respectively. These present results indicate that CH-stretching first, combination band and second overtone regions can serve as diagnostic markers for gastric cancer.

[Growth hormone enhances regeneration of cavernous nerves after their transplantation in rats].

OBJECTIVE: To investigate the effect of growth hormone (GH) on penile erection after reconstruction of cavernous nerves using sural nerve as an interposition nerve graft in rats. METHODS: Twenty-four male Sprague-Dawley rats (3-4 ms of age and 300-400 g in weight) were randomly divided into 2 groups: nerve graft group and GH group, each electrostimulated to determine the erectile potency 2 and 4 months after nerve graft (followed by hypodermic GH injection). The nNOS-positive nerve fibers in the corpora cavemosa were examined by streptavidin-peroxidase immunohistochemistry technique (SP method). Image analysis was used to calculate the area stained in pixel. RESULTS: Electrostimulation at 2 months produced 31.25% of erections in the GH group but none in the grafted rats. There was a significant difference in the erection rate produced by electrostimulation between the two groups at 2 months (P < 0.05). The pixel of the expression of nNOS-positive nerve fibers in the GH group (38971 +/- 7692) was also greater than that of the graft group (16538 +/- 3179, P < 0.05). At 4 months, 43.75% of the graft group and 75% of the GH group produced erections upon electrostimulation, with no significant difference between the two groups (P > 0.05). The pixels of the expression of nNOS-positive nerve fibers were 79276 +/- 12,021 and 91348 +/- 18965, respectively (P > 0.05). CONCLUSION: GH can accelerate the regeneration of cavernous nerves after bilateral nerve grafting, and GH administration may present a new physiological approach to the treatment of erectile dysfunction after radical pelvic surgery.

[Effects on erectile function of transplanted major pelvic ganglion into the corpus cavernosum of adult rats with bilateral cavernous nerve injury].

OBJECTIVE: To investigate the effects on erectile function of transplanted major pelvic ganglion into the corpus cavernosum of adult male rats undergoing transection of bilateral cavernous nerves. METHODS: Twenty-six male Sprague-Dawley rats (3 - 4 month-old and 300 - 400 g/each) were divided into 2 groups: experimental group (transection of bilateral cavernous nerves and transplantation of left ganglion into left crus of penis, n = 16) and control group (transection of bilateral cavernous nerves only, n = 10). Erectile function was measured by injecting APO, and intracavernous pressure was measured 1 and 3 months afterwards by electric-stimulating the right major pelvic ganglion or the left crus. Half animals in each group were sacrificed 1 and 3 months afterwards for detecting nNOS-containing nerve fibers of corpus cavernosum. Electron microscopy of the implanted area was performed to assess neuronal survival. RESULTS: Both of the two groups have no erectile response to APO injection. Electrostimulation on the right major pelvic ganglion and left crus failed to produce erection in experimental group. The mean pressure changes in the two groups, measured by stimulating the left crus, were (9.41 +/- 3.20) and (4.16 +/- 2.58) cmH(2)O 1 month afterwards, and (13.67 +/- 4.18) and (5.09 +/- 2.74) cmH(2)O 3 months afterwards, respectively (P < 0.05). An increased number of nNOS-containing nerve fibers in left crus was detected in experimental group 1 and 3 months later, compared with control one (218.7 +/- 24.5, 18.0 +/- 3.7; 183.2 +/- 19.7, 19.0 +/- 3.8; P < 0.05). Ultrastructure examination by transmission electron microscope confirmed the survival of the implanted ganglion. CONCLUSION: Major pelvic ganglion can survive in the corpus cavernosum, and it has significant effects on the number of nNOS-containing nerve fibers and the alteration of intracavernous pressure.

[Restoration of erectile function by reconstructing cavernous nerves with sural nerve grafts].

OBJECTIVE: To investigate the restoration of erectile function by reconstructing cavernous nerves with sural nerve grafts. METHODS: Forty-eight male Sprague-Dawley rats(3-4 m old and 300-400 g) were randomly divided into three groups: the sham-operated group (n = 16) underwent pelvic exploration without transection of the cavernous nerve; the nerve ablation group (n = 16) had a 5 mm segment of the cavernous nerve excised bilaterally; the graft group (n = 16) had a 5 mm segment of the cavernous nerve excised bilaterally, followed by immediate microsurgical reconstruction with an interposition graft of the sural nerve. The cavernous nerves of each group were electrostimulated to determine their potency after 2 and 4 months. And fluorescent retrograde-transported material Fluoro-Gold(FG) was injected into the penis. FG-labeled neuron cells in whole mounts of major pelvic ganglions were observed five days after injection. RESULTS: Electrical stimulation produced no erection in either the nerve ablation or the graft group, but 100% erection in the sham-operated group after 2 months. The numbers of FG-labeled neurons significantly differed between the nerve ablation group and the graft group. After 4 months erection examination showed statistical significance in the difference between the graft group and the nerve ablation group(P < 0.05). The FG-labeled neurons in the graft group significantly differed from those in the ablation (P < 0.05), and almost reached the level of the sham-operated(P < 0.05). CONCLUSION: Cavernous nerve grafting can successfully restore erectile dysfunction in rats after surgical injury.