[Research progress of three-dimensional bioprinting technology in auricle repair and reconstruction].

Objective: To review the research progress on the application of three-dimensional (3D) bioprinting technology in auricle repair and reconstruction. Methods: The recent domestic and international research literature on 3D printing and auricle repair and reconstruction was extensively reviewed, and the concept of 3D bioprinting technology and research progress in auricle repair and reconstruction were summarized. Results: The auricle possesses intricate anatomical structure and functionality, necessitating precise tissue reconstruction and morphological replication. Hence, 3D printing technology holds immense potential in auricle reconstruction. In contrast to conventional 3D printing technology, 3D bioprinting technology not only enables the simulation of auricular outer shape but also facilitates the precise distribution of cells within the scaffold during fabrication by incorporating cells into bioink. This approach mimics the composition and structure of natural tissues, thereby favoring the construction of biologically active auricular tissues and enhancing tissue repair outcomes. Conclusion: 3D bioprinting technology enables the reconstruction of auricular tissues, avoiding potential complications associated with traditional autologous cartilage grafting. The primary challenge in current research lies in identifying bioinks that meet both the mechanical requirements of complex tissues and biological criteria.

DLGAP5 triggers proliferation and metastasis of bladder cancer by stabilizing E2F1 via USP11.

Bladder cancer (BLCA) is one of the most widespread malignancies worldwide, and displays significant tumor heterogeneity. Understanding the molecular mechanisms exploitable for treating aggressive BLCA represents a crucial objective. Despite the involvement of DLGAP5 in tumors, its precise molecular role in BLCA remains unclear. BLCA tissues exhibit a substantial increase in DLGAP5 expression compared with normal bladder tissues. This heightened DLGAP5 expression positively correlated with the tumor's clinical stage and significantly affected prognosis negatively. Additionally, experiments conducted in vitro and in vivo revealed that alterations in DLGAP5 expression notably influence cell proliferation and migration. Mechanistically, the findings demonstrated that DLGAP5 was a direct binding partner of E2F1 and that DLGAP5 stabilized E2F1 by preventing the ubiquitination of E2F1 through USP11. Furthermore, as a pivotal transcription factor, E2F1 fosters the transcription of DLGAP5, establishing a positive feedback loop between DLGAP5 and E2F1 that accelerates BLCA development. In summary, this study identified DLGAP5 as an oncogene in BLCA. Our research unveils a novel oncogenic mechanism in BLCA and offers a potential target for both diagnosing and treating BLCA.

Pectolinarigenin inhibits bladder urothelial carcinoma cell proliferation by regulating DNA damage/autophagy pathways.

Pectolinarigenin (PEC), an active compound isolated from traditional herbal medicine, has shown potential anti-tumor properties against various types of cancer cells. However, its mechanism of action in bladder cancer (BLCA), which is one of the fatal human carcinomas, remains unexplored. In this study, we first revealed that PEC, as a potential DNA topoisomerase II alpha (TOP2A) poison, can target TOP2A and cause significant DNA damage. PEC induced G2/M phase cell cycle arrest via p53 pathway. Simultaneously, PEC can perform its unique function by inhibiting the late autophagic flux. The blocking of autophagy caused proliferation inhibition of BLCA and further enhanced the DNA damage effect of PEC. In addition, we proved that PEC could intensify the cytotoxic effect of gemcitabine (GEM) on BLCA cells in vivo and in vitro. Summarily, we first systematically revealed that PEC had great potential as a novel TOP2A poison and an inhibitor of late autophagic flux in treating BLCA.

Melatonin inhibits bladder tumorigenesis by suppressing PPARγ/ENO1-mediated glycolysis.

Melatonin is a well-known natural hormone, which shows a potential anticancer effect in many human cancers. Bladder cancer (BLCA) is one of the most malignant human cancers in the world. Chemoresistance is an increasingly prominent phenomenon that presents an obstacle to the clinical treatment of BLCA. There is an urgent need to investigate novel drugs to improve the current clinical status. In our study, we comprehensively explored the inhibitory effect of melatonin on BLCA and found that it could suppress glycolysis process. Moreover, we discovered that ENO1, a glycolytic enzyme involved in the ninth step of glycolysis, was the downstream effector of melatonin and could be a predictive biomarker of BLCA. We also proved that enhanced glycolysis simulated by adding exogenous pyruvate could induce gemcitabine resistance, and melatonin treatment or silencing of ENO1 could intensify the cytotoxic effect of gemcitabine on BLCA cells. Excessive accumulation of reactive oxygen species (ROS) mediated the inhibitory effect of melatonin on BLCA cells. Additionally, we uncovered that PPARγ was a novel upstream regulator of ENO1, which mediated the downregulation of ENO1 caused by melatonin. Our study offers a fresh perspective on the anticancer effect of melatonin and encourages further studies on clinical chemoresistance.

MYBL2 promotes proliferation and metastasis of bladder cancer through transactivation of CDCA3.

The transcription factor MYB proto-oncogene like 2 (MYBL2) is critical in regulating gene expression and tumorigenesis. However, the biological function of MYBL2 in bladder cancer (BLCA) remains to be elucidated. Here, we first revealed that MYBL2 was elevated in BLCA tissues and significantly correlated with clinicopathological parameters and cancer-specific survival in BLCA patients. Phenotypic assays showed that MYBL2 deficiency suppressed the proliferation and migration of BLCA cells in vitro and in vivo, whereas MYBL2 overexpression contributed to the opposite phenotype. Mechanistically, MYBL2 could bind to the promoter of its downstream target gene cell division cycle-associated protein 3 (CDCA3) and transactivate it, which in turn promoted the malignant phenotype of BLCA cells. Further investigations revealed that MYBL2 interacted with forkhead box M1 (FOXM1) to co-regulate the transcription of CDCA3. In addition, MYBL2/FOXM1 and CDCA3 might activate Wnt/ß-catenin signaling, thereby promoting the malignant phenotype of BLCA cells. In conclusion, the current study identifies MYBL2 as an oncogene in BLCA. MYBL2 can accelerate the proliferation and metastasis of BLCA through the transactivation of CDCA3.

The Role of Long Non-Coding RNAs in Epithelial-Mesenchymal Transition-Related Signaling Pathways in Prostate Cancer.

Prostate cancer (PCa) is one of the most common male malignancies with frequent remote invasion and metastasis, leading to high mortality. Epithelial-mesenchymal transition (EMT) is a fundamental process in embryonic development and plays a key role in tumor proliferation, invasion and metastasis. Numerous long non-coding RNAs (lncRNAs) could regulate the occurrence and development of EMT through various complex molecular mechanisms involving multiple signaling pathways in PCa. Given the importance of EMT and lncRNAs in the progression of tumor metastasis, we recapitulate the research progress of EMT-related signaling pathways regulated by lncRNAs in PCa, including AR signaling, STAT3 signaling, Wnt/ß-catenin signaling, PTEN/PI3K/AKT signaling, TGF-ß/Smad and NF-κB signaling pathways. Furthermore, we summarize four modes of how lncRNAs participate in the EMT process of PCa via regulating relevant signaling pathways.

Friend or Foe? The Roles of Antioxidants in Acute Lung Injury.

Acute lung injury (ALI) is an acute hypoxic respiratory insufficiency caused by various intra- and extra-pulmonary injury factors. The oxidative stress caused by excessive reactive oxygen species (ROS) produced in the lungs plays an important role in the pathogenesis of ALI. ROS is a "double-edged sword", which is widely involved in signal transduction and the life process of cells at a physiological concentration. However, excessive ROS can cause mitochondrial oxidative stress, leading to the occurrence of various diseases. It is well-known that antioxidants can alleviate ALI by scavenging ROS. Nevertheless, more and more studies found that antioxidants have no significant effect on severe organ injury, and may even aggravate organ injury and reduce the survival rate of patients. Our study introduces the application of antioxidants in ALI, and explore the mechanisms of antioxidants failure in various diseases including it.

The inhibitory effect of silencing CDCA3 on migration and proliferation in bladder urothelial carcinoma.

BACKGROUND: CDCA3 is an important component of the E3 ligase complex with SKP1 and CUL1, which could regulate the progress of cell mitosis. CDCA3 has been widely identified as a proto-oncogene in multiple human cancers, however, its role in promoting human bladder urothelial carcinoma has not been fully elucidated. METHODS: Bioinformatic methods were used to analyze the expression level of CDCA3 in human bladder urothelial carcinoma tissues and the relationship between its expression level and key clinical characteristics. In vitro studies were performed to validate the specific functions of CDCA3 in regulating cell proliferation, cell migration and cell cycle process. Alterations of related proteins was investigated by western blot assays. In vivo studies were constructed to validate whether silencing CDCA3 could inhibit the proliferation rate in mice model. RESULTS: Bioinformatic analysis revealed that CDCA3 was significantly up-regulated in bladder urothelial carcinoma samples and was related to key clinical characteristics, such as tumor grade and metastasis. Moreover, patients who had higher expression level of CDCA3 tend to show a shorter life span. In vitro studies revealed that silencing CDCA3 could impair the migration ability of tumor cells via down-regulating EMT-related proteins such as MMP9 and Vimentin and inhibit tumor cell growth via arresting cells in the G1 cell cycle phase through regulating cell cycle related proteins like p21. In vivo study confirmed that silencing CDCA3 could inhibit the proliferation of bladder urothelial carcinoma cells. CONCLUSIONS: CDCA3 is an important oncogene that could strengthen the migration ability of bladder urothelial carcinoma cells and accelerate tumor cell growth via regulating cell cycle progress and is a potential biomarker of bladder urothelial carcinoma.

The inhibitory effect of melatonin on human prostate cancer.

Prostate cancer (PCa) is one of the most commonly diagnosed human cancers in males. Nearly 191,930 new cases and 33,330 new deaths of PCa are estimated in 2020. Androgen and androgen receptor pathways played essential roles in the pathogenesis of PCa. Androgen depletion therapy is the most used therapies for primary PCa patients. However, due to the high relapse and mortality of PCa, developing novel noninvasive therapies have become the focus of research. Melatonin is an indole-like neurohormone mainly produced in the human pineal gland with a prominent anti-oxidant property. The anti-tumor ability of melatonin has been substantially confirmed and several related articles have also reported the inhibitory effect of melatonin on PCa, while reviews of this inhibitory effect of melatonin on PCa in recent 10 years are absent. Therefore, we systematically discuss the relationship between melatonin disruption and the risk of PCa, the mechanism of how melatonin inhibited PCa, and the synergistic benefits of melatonin and other drugs to summarize current understandings about the function of melatonin in suppressing human prostate cancer. We also raise several unsolved issues that need to be resolved to translate currently non-clinical trials of melatonin for clinic use. We hope this literature review could provide a solid theoretical basis for the future utilization of melatonin in preventing, diagnosing and treating human prostate cancer. Video abstract.

CTHRC1 Is a Prognostic Biomarker and Correlated With Immune Infiltrates in Kidney Renal Papillary Cell Carcinoma and Kidney Renal Clear Cell Carcinoma.

Kidney renal clear cell carcinoma (KIRC) and kidney renal papillary cell carcinoma (KIRP) are the most common RCC types. RCC has high immune infiltration levels, and immunotherapy is currently one of the most promising treatments for RCC. Collagen triple helix repeat containing 1 (CTHRC1) is an extracellular matrix protein that regulates tumor invasion and modulates the tumor microenvironment. However, the association of CTHRC1 with the prognosis and tumor-infiltrating lymphocytes of KIRP and KIRC has not been reported. We examined the CTHRC1 expression differences in multiple tumor tissues and normal tissues via exploring TIMER, Oncomine, and UALCAN databases. Then, we searched the Kaplan-Meier plotter database to evaluate the correlation of CTHRC1 mRNA level with clinical outcomes. Subsequently, the TIMER platform and TISIDB website were chosen to assess the correlation of CTHRC1 with tumor immune cell infiltration level. We further explored the causes of aberrant CTHRC1 expression in tumorigenesis. We found that CTHRC1 level was significantly elevated in KIRP and KIRC tissues relative to normal tissues. CTHRC1 expression associates with tumor stage, histology, lymph node metastasis, and poor clinical prognosis in KIRP. The CTHRC1 level correlates to tumor grade, stage, nodal metastasis, and worse survival prognosis. Additionally, CTHRC1 is positively related to different tumor-infiltrating immune cells in KIRP and KIRC. Moreover, CTHRC1 was closely correlated with the gene markers of diverse immune cells. Also, high CTHRC1 expression predicted a worse prognosis in KIRP and KIRC based on immune cells. Copy number variations (CNV) and DNA methylation might contribute to the abnormal upregulation of CTHRC1 in KIRP and KIRC. In conclusion, CTHRC1 can serve as a biomarker to predict the prognosis and immune infiltration in KIRP and KIRC.

Identification of 9 key genes and small molecule drugs in clear cell renal cell carcinoma.

Clear cell renal cell carcinoma (ccRCC) is a heterogeneous tumor that the underlying molecular mechanisms are largely unclear. This study aimed to elucidate the key candidate genes and pathways in ccRCC by integrated bioinformatics analysis. 1387 differentially expressed genes were identified based on three expression profile datasets, including 673 upregulated genes and 714 downregulated genes. Then we used weighted correlation network analysis to identify 6 modules associated with pathological stage and grade, blue module was the most relevant module. GO and KEGG pathway analyses showed that genes in blue module were enriched in cell cycle and metabolic related pathways. Further, 25 hub genes in blue module were identified as hub genes. Based on GEPIA database, 9 genes were associated with progression and prognosis of ccRCC patients, including PTTG1, RRM2, TOP2A, UHRF1, CEP55, BIRC5, UBE2C, FOXM1 and CDC20. Then multivariate Cox regression showed that the risk score base on 9 key genes signature was a clinically independent prognostic factor for ccRCC patients. Moreover, we screened out several new small molecule drugs that have the potential to treat ccRCC. Few of them were identified as biomarkers in ccRCC. In conclusion, our research identified 9 potential prognostic genes and several candidate small molecule drugs for ccRCC treatment.

[Effects of different core temperatures after heat strike on serum inflammatory cytokines and multiple organ dysfunction syndrome in rats].

OBJECTIVE: To observe the effect of different core temperatures (Tc) after heat strike on serum inflammatory cytokines and multiple organ dysfunction syndrome (MODS) in rat. METHODS: 120 male Sprague-Dawley (SD) rats were randomly divided into normal control group (n = 30) and heat strike group (n = 90). The rats in heat strike group were put into simulated thermal climate animal module after adaptive training. The module temperature was raised to 39 centigrade in 30 minutes with 65% humidity. The rats ran simultaneously at 15 m/min, on the slope of 0 degree angle, 8 minutes each time, 2 minutes interval, and the heat strike time was 90 minutes. After the rats came out of the module, rectal temperature, which was Tc value, was recorded. The rats died or Tc < 41 centigrade during the experiment were excluded, the remaining 73 rats were divided into three subgroups: 41.0-41.9 centigrade (n = 38), 42.0-42.9 centigrade (n = 26), and ≥43.0 centigrade (n = 9). The rats in the normal control group were reared at temperature of (25±2) centigrade, and humidity of (55±5)%. At 0 hour and 24 hours after the rats came out of the module, femoral artery blood was collected to determine serum interleukins (IL-1α, IL-1ß, IL-17), tumor necrosis factor-α(TNF-α) andγ-interferon (IFN-γ) by enzyme-linked immunosorbent assay (ELISA). The cardiac troponin I (cTnI), MB isoenzyme of creatine kinase (CK-MB), serum creatinine (SCr), blood urea nitrogen (BUN), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) levels were determined by automatic biochemical analyzer. The incidence of MODS and the number of accumulative organs within 24 hours of the rats in different Tc of heat strike group were compared and analyzed. RESULTS: The serum inflammatory cytokines and biochemical parameters at 0 hour after heat strike were significant higher than those of the normal control group, and showed a time dependence. Further analysis showed that the inflammatory response and organ dysfunction in rats were increased gradually with the increase in Tc of rats. Compared with the normal control group, at 24 hours after heat strike, inflammatory cytokines in Tc≥43.0 centigrade rats were increased obviously [IL-1α (ng/L): 13.56±2.07 vs. 2.24±0.62, IL-1ß (ng/L): 17.11±1.90 vs. 7.40±1.52, IL-17 (ng/L): 17.00±1.41 vs. 6.00±1.78, TNF-α (ng/L): 16.78±1.79 vs. 7.27±1.74, IFN-γ (ng/L): 21.11±2.09 vs. 10.43±2.31], and the biochemical parameters were also increased obviously [cTnI (ng/L): 50.78±6.67 vs. 20.53±3.09, CK-MB (U/L): 62.89±3.82 vs. 22.00±3.01, SCr (µmol/L): 149.22±4.35 vs. 92.53±8.32, BUN (nmol/L): 55.22±1.99 vs. 19.10±2.02, ALT (U/L): 388.33±4.97 vs. 100.23±10.61, AST (U/L): 361.22±6.53 vs. 97.67±10.54, all P < 0.01]. The incidence of MODS within 24 hours in the heat strike group was 54.79% (40/73), and the higher the Tc, the higher the incidence of MODS, and the more insulted organs [the incidence of MODS in 41.0-41.9 centigrade, 42.0-42.9 centigrade, and ≥43.0 centigrade subgroups was 36.84% (14/38), 65.38% (17/26), 100.00% (9/9), and the organ involvement rate was 12.17% (37/304), 23.08% (48/208), and 48.61% (35/72), respectively, when 8 organs or systems were calculated for each rat, both P < 0.01]. CONCLUSIONS: The higher the Tc of heat strike rats, the stronger the inflammatory reaction and the more serious the damage of tissue, and the more extensive damage of the organs.

[Protective effect of mild hypothermia at different starting times on organ function in patient with exertional heat stroke].

OBJECTIVE: To investigate the protective effect of mild hypothermia at different starting times on the physiological functions of the viscera of exertional heat stroke (EHS). METHODS: A prospective randomized controlled trial was conducted. EHS patients admitted to intensive care unit of the 159th Hospital of People's Liberation Army and the First Affiliated Hospital of Zhengzhou University from June 2015 to June 2017 were enrolled. The patients were divided into 2, 4, 6 hours start hypothermia treatment groups according to the random number table method, the mild hypothermia was initiated at 2, 4 and 6 hours after the disease onset respectively, and the methods were the same in each group. After treatment of 2, 12, 24 hours, the venous blood in the three groups was collected to detect serum cardiac troponin I (cTnI) with chemiluminescence method, MB isoenzyme of creatine kinase (CK-MB) with immunosuppressive method, creatinine (Cr) with creatine oxidase method, ß2-microglobulin (ß2-MG) with turbidimetry, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) with enzyme method. Multiple organ dysfunction syndrome (MODS) within 24 hours after treatment was recorded. Linear regression analysis of the correlation between mild hypothermia start-up time and MODS was done. RESULTS: Ninety-three cases of EHS were included, with 32, 31 and 30 patients in 2, 4, 6 hours start treatment groups respectively. There were no significant differences in gender, age, core temperature, onset time to admission, Glasgow coma scale (GCS), acute physiology and chronic health evaluation system II (APACHE II) score at admission among the three groups. There were no significant differences in the levels of serum cTnI, CK-MB, Cr, ß2-MG, ALT and AST at 2 hours after treatment. But with the prolongation of the treatment time, all indicators gradually increased. And the earlier start of the mild hypothermia, the less significant of the above indexes. All indexes in 2 hours start treatment group were significantly lower than those of 2 hours and 6 hours start treatment groups at 24 hours after treatment [cTnI (ng/L): 49.53±9.25 vs. 56.52±10.05, 64.57±11.21; CK-MB (U/L): 51.47±11.83 vs. 57.87±7.43, 64.40±7.93; Cr (µmol/L): 140.97±11.33 vs. 148.16±10.39,155.57±8.65; ß2-MG (mg/L): 10.28±1.46 vs. 11.58±2.13, 12.93±1.98; ALT (U/L): 248.53±75.47 vs. 341.42±129.58, 425.77±101.23; AST (U/L): 197.25±42.59 vs. 292.81±58.49, 351.20±60.41, all P < 0.05]. There was significant difference in the incidence of MODS in 2, 4, 6 hours start treatment groups [43.75% (14/32), 64.52% (20/31), 80.08% (24/30), χ2 = 8.761, P = 0.013]. Linear regression analysis showed that the earlier onset time of mild hypothermia, the lower incidence of MODS (R2 = 0.915, P = 0.013). CONCLUSIONS: The application of mild hypothermia in 2 hours can effectively protect the physiological function of EHS organs and reduce the incidence of MODS.

[Rapid diagnosis of common pathogenic bacteria infection in newborn infants by 16SrDNA oligonucleotide array].

OBJECTIVE: The rapid identification of pathogenic bacteria is important for earlier effective patient management and antimicrobial therapy, especially for the infant patient, whose immunological system is not fully developed. However conventional microbiogical techniques of bacterial identification, culture and isolation of pathogenic bacteria, identification by biochemistry and serological assay, are time-consuming and require intensive labor. On the basis of special gene sequence, PCR provides simple and rapid way to identify bacteria. But it is difficult to identify all of bacteria species which are suspicious of pathogenic agents. Oligonucleotide arrays provide a powerful tool for parallel detection of target genes. The objective of this study was to test a reverse oligonucleotide assay, which hybridize with the PCR product of 16SrDNA using a pair of universal primers, to rapidly identify common infant pathogenic bacteria. METHODS: By comparison and analysis of the 16SrDNA sequences of common pathogenic bacteria, a region, which has numerous sequence variations and flanked by highly conserved sequences, was found. A pair of universal primers was designed according to its flanking conservative sequence, and a set of probes specially targeting to eight species of infant pathogenic bacteria, including staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae, Streptococcus faecalis, Hemophilus influenzae, Enterobacter cloacae, Escherichia coli, and Acinetobacter baumannii,according to the variable sequences. The probes were fixed on the nylon membrane with positive electricity, and hybridized them with the products of PCR using the universal primers. RESULTS: The universal primers could amplify the target sequence from bacteria including the eight common infant pathogenic bacteria and Staphylococcus epidermidis, Enterobacter aerogenes, Streptococcus pneumoniae,beta-hemolytic streptococcus, Neisseria meningitides, Citrobacter freundii, Bacillus subtilis, and Salmonella infantis,but could not amplify rotavirus and human DNA as control. The results showed that the oligonucleotide array could specially hybridize with the eight bacteria to be examined and could not hybridize with other bacteria. The lowest concentration of DNA (product of PCR) for oligonucleotide array was about 25 ng/ml. The results proved that the probes are highly selective and the oligonucleotide arrays could parallelly detect the eight common infant pathogenic bacteria. The results suggested that the oligonucleotide array system was able to identify the eight common infant pathogenic bacteria from clinical specimens and the results were the same as identified by automated bacterial detection machine. From the further experiments, the oligonucleotide array system could directly diagnose the common infant pathogenic bacteria from the broths of samples culture. CONCLUSIONS: Despite limited number of identifiable bacteria and lack of information on antibiotic susceptibility of bacteria, the reverse oligonucleotide assay system, which contains amplification of the segment of 16rDNA from samples using the universal primers and parallel detection of PCR products using specific probes, is an effective method to rapidly identify the eight common infant pathogenic bacteria.