[Key updates and interpretation of the ninth version of AJCC staging system for neuroendocrine tumors of the stomach].

The cancer staging system of the American Joint Committee on Cancer (AJCC) is the most widely used clinical basis for tumor staging. In October 2023, AJCC released the staging system (ninth version) for the neuroendocrine tumors of stomach (NET), which has been implemented in January 2024. The ninth version of NET staging system mainly updated the histopathologic classification, diagnosis and staging methods, clinical and pathological staging, prognosis grade, tumor and non-tumor prognostic features. The update and implementation of the staging system provide a more detailed reference for the accurate diagnosis, staging and precise treatment of gastric neuroendocrine tumors. Moreover, it is convenient for clinicians to carry out clinical practice. The purpose of our article is to provide a high-level overview of the major changes in AJCC staging system (version 9) for gastric NET based on the latest evidence-based medical research.

[Analysis of factors associated with proximal junctional kyphosis after surgery for Lenke type 5 adolescent idiopathic scoliosis].

Objective: To investigate the risk factors associated with the development of proximal junctional kyphosis (PJK) after posterior spinal fusion for in children with Lenke type 5 adolescent idiopathic scoliosis (AIS). Methods: It was a retrospective case-control study that included medical records of 98 children with Lenke type 5 AIS who underwent posterior orthopedic surgery under general anesthesia at the Honghui Hospital Affiliated to Xi'an Jiaotong University from January 2013 to December 2018. There were 23 males and 75 females with a mean age of (14.5±2.2) years (10-18 years). Patients were divided into PJK and non-PJK groups according to whether the posterior junctional angle (PJA) was greater than 10° and increased for more than 10° from the preoperative period at the the last follow-up. Univariate analysis was used to analyze the correlation of general data of the children with occurrence of PJK after the operation. Multivariate logistic regression analysis was used to analyze the risk factors of postoperative PJK. Results: There were 35 cases in the PJK group and 63 cases in the non-PJK group. The PJK and non-PJK groups were followed up for (35.6±7.3) months and (36.4±7.5) months, respectively, and the difference was not statistically significant (P=0.637). There was no statistically significant difference between the two groups in general data such as gender, age, and body mass index (all P>0.05), while there were statistically significant differences between the two groups in upper instrumented vertebrea (UIV) location and junctional area posterior ligamentous complex (PLC) injury (all P<0.05). The results of univariate analysis showed that UIV location at T10-T12, junctional area PLC injury, preoperative coronal thoracic curve (TC), preoperative and final follow-up PJA, and preoperative and final follow-up pelvic incidence-lumbarlordosis (PI-LL) were correlated with postoperative PJK (OR=2.50, 5.37, 0.92, 1.12, 1.32, 1.06, 3.35, all P<0.05). Multifactorial logistic regression analysis showed that UIV located at T10-T12 (OR=2.346, 95%CI: 1.582-3.481, P=0.001), junctional area PLC injury (OR=5.112, 95%CI: 1.283-20.418, P=0.023) and last follow-up PI-LL (OR=1.826, 95%CI: 1.558-24.745, P=0.012) were risk factors for the occurrence of postoperative PJK in children with Lenke type 5 AIS. Conclusions: Postoperative UIV fixation to the thoracolumbar segment, PLC injury in the junctional area and excessive postoperative PI-LL in children with Lenke type 5 AIS may be the risk factors for the occurrence of PJK after the operation. It is suggested that avoidance of UIV selection to the thoracolumbar segment, intraoperative protection of the PLC located near the UIV and restoration of a good PI-LL relationship may reduce the incidence of PJK.

[Expression of hepatocyte nuclear factor 4γ in gastric carcinoma and its role in cell proliferation and stemness].

Objective: To explore the role and molecular mechanism of hepatocyte nuclear factor 4γ (HNF4γ) in proliferation and stemness of gastric cancer. Methods: A total of 102 cases of paraffin-embedded gastric cancer tissues and matched adjacent gastric tissues and 42 cases of fresh-frozen tissues derived from gastric patients who received radical gastrectomy were collected from the First Affiliated Hospital of Zhengzhou University between 2012 to 2015. The expression of HNF4γ was tested by immunohistochemical staining, quantitative real-time polymerase chain reaction (qRT-PCR). HNF4γ overexpressed (AGS-HNF4γ) and shRNA silenced (HGC27-shHNF4γ) gastric cell lines were established. The effects of HNF4γ on cell proliferation and stemness were verified by XTT, clone formation and sphere formation assay. The expression of CD44 was detected by western blot. Results: The mRNA expression level of HNF4γ in fresh-frozen gastric cancer tissue was (12.43±2.702), which was significantly higher than (3.639±1.109) in normal tissue (P<0.001). The high protein expression rate of HNF4γ in paraffin-embedded gastric cancer tissues was 41.2% (42/102), which was significantly higher than 8.8% (9/102) in normal gastric mucosa tissue (P< 0.001). The protein expression of HNF4γ was closely related to the tumor differentiation, infiltration depth, lymph node metastasis and tumor stage (P<0.05). The median survival interval of patients with HNF4γ high expression was 25 months, the 3-year survival rate was 4.8% (2/42), significantly lower than 38 months and 51.7% (31/60) of patients with normal HNF4γ expression (P<0.001). The proliferation and CD44 protein expression of AGS-HNF4γ cells were significantly higher than those of the AGS-Vector cells. The number of clone formation, sphere formation rate of AGS-HNF4γ cells were 243.5±24.5 and (83.5±3.9)%, significantly higher than 81.0±16.0 and (21.8±5.6)% of AGS-Vector cells (P=0.030 and P=0.010, respectively). The proliferation and CD44 protein expression of HGC27-shHNF4 cells were significantly lower than those of the HGC27-vector cells. The number of clone formation, sphere formation rate of HGC27-shHNF4 cells were 26.0±1.0 and (20.8±8.4)%, significantly higher than 83.5±4.5 and (72.5±4.8)% of HGC27-vector cells (P=0.006 and P=0.030, respectively). Conclusions: HNF4γ is upregulated in the gastric cancer tissues and related with the poor prognosis of patients with gastric cancer. Overexpression of HNF4γ promotes the proliferation and remains the stemness of gastric cancer cells by upregulating the expression of CD44.

Correlation between MRAS gene polymorphism and atherosclerosis.

OBJECTIVE: The aim of this study was to explore the correlation between muscle RAS oncogene homolog (MRAS) gene polymorphism and the onset risk of atherosclerosis (AS). PATIENTS AND METHODS: A total of 135 AS patients diagnosed and treated in our hospital from November 2017 to October 2018 were randomly enrolled in the observation group. Meanwhile, 150 healthy adults were selected as control group. Venous blood was withdrawn from all the subjects, and DNAs were extracted. MRAS gene loci rs9818870 and rs3755751 were analyzed by the multiplex SNaPshot method, and their correlations with the onset risk of AS were explored. RESULTS: No statistically significant differences in the frequencies at gene loci were observed between the two groups (p>0.05). Subjects with genotype TT at rs9818870 exhibited significantly higher risk of AS (p=0.041<0.05). The recessive model of rs9818870 (GG + AG/AA) in AS patients with coronary heart disease was correlated with AS (p=0.048<0.05). Similarly, the dominant model of rs3755751 (TT/TC+CC) in those with hypertension was associated with AS (p=0.027<0.05). CONCLUSIONS: MRAS gene is correlated with the onset of AS to a certain degree.

[Robotic hilar cholangiocarcinoma radical resection compared with laparotomy in prognosis].

Objective: To compare the long term and short term outcomes between robotic and open surgery for hilar cholangiocarcinoma radical resection. Methods: This is a single-center and retrospective case-control study. Patients underwent hilar cholangiocarcinoma radical resection between January 2016 and December 2016 at Department of Hepatobiliary Surgery of the General Hospital of the Chinese People's Liberation Army were included. Safety, effectiveness and long-term prognosis of tumors were evaluated. Patients were divided into robotic hilar cholangiocarcinoma radical surgery group (robotic group, n=16) and open hepatic hilar cholangiocarcinoma radical surgery group (open group, n=31) . All cases were confirmed by pathology histological. Age, gender, histology, resection margin status, extent of surgical resection, disease-free survival (DFS) , and overall survival (OS) were retrospectively collected and analyzed.In the follow-up cohort, the primary outcome was patient death and the secondary outcome was tumor recurrence. Continuous variables were expressed as means and medians and were compared using the Student t test if normally distributed or Wilcoxon-Mann-Whitney test for nonparametric variables. Discrete variables were expressed as frequency and percentages and χ(2) or Fisher exact test, when appropriate, were used for comparisons. Kaplan-Meier curves were used to calculate the probability of survival and comparisons were performed using log-rank test. Results: In this study, compared with the open group, the robotic group had a longer operation time ( (338±71) minutes vs. (256±56) minutes, t=4.251, P=0.001) , but the intraoperative blood loss was less (100 ml vs. 200 ml, Z=121.50, P=0.040) , the gastric tube removal time was earlier (3 days vs. 4 days, Z=136.0, P=0.011) , and the postoperative hospital stay was shorter (9 days vs. 12 days, Z=144.50, P=0.040) , and the difference was statistically significant.There was no significant difference in the blood transfusion rate, R0 resection rate, and tumor size between the two groups.The recurrence rates in the robotic group and open surgery were 53.3% and 67.0%, respectively (χ(2)=1.04, P=0.307) .The median survival time of the robotic group and the open group was 22.0 months and 25.0 months. There was no significant difference in the overall survival rate between the two groups (P>0.05) . Conclusion: Compared with laparotomy, robotic HCC radical resection could have an equivalence or non-inferiority approach with acceptable long-term outcome.

The value of turbo spin-echo diffusion-weighted imaging apparent diffusion coefficient in the diagnosis of temporal bone cholesteatoma.

AIM: To explore the value of the applying the apparent diffusion coefficient (ADC) to the quantitative diagnosis of temporal bone cholesteatoma. MATERIALS AND METHODS: Seventy-one patients clinically suspected of temporal bone cholesteatoma were enrolled prospectively. These patients underwent ear magnetic resonance imaging (MRI) and turbo spin-echo diffusion-weighted imaging (TSE-DWI) using a Philips Ingenia 3 T superconductive MRI system, and their ADCs were measured. Subsequently, all enrolled patients underwent surgery within 15 days of the MRI. Using receiver operating characteristic (ROC) curve analysis, the optimal threshold and diagnostic performance for diagnosing temporal bone cholesteatoma were determined. Logistic regression modelling was used to combine ADCs and T1-weighted imaging (T1WI) sequences and calculate the combined diagnostic performance. RESULTS: Based on the pathology results, patients were categorised into the cholesteatoma group (CS group, n=43) and the non-cholesteatoma group (NCS group, n=28). In the CS group, ADCs were significantly lower (mean, 0.87±0.31×10-3 mm2/s) than in the NCS group (mean, 1.87±0.49×10-3 mm2/s; p<0.001); the area under the ROC curve (AUC) was 0.915. Based on the optimal threshold of ADC, ≤1.23×10-3 mm2/s, the diagnostic performance was high; the sensitivity and specificity for diagnosing cholesteatoma were 95.35% and 85.71%, respectively. By combining the ADC and T1WI sequence, AUC increased to 0.953, and the sensitivity and specificity were 90.7% and 96.43%, respectively. CONCLUSION: ADC quantitative analysis has a high value in the preoperative diagnosis of cholesteatoma, and the combination of ADC and T1WI sequences can improve the diagnostic accuracy and specificity and thereby facilitate clinically effective diagnosis.

Characterization of facial asymmetry in skeletal Class III malocclusion and its implications for treatment.

Jaw deviation is frequently seen in Class III patients. The aim of the study was to investigate asymmetric features of skeletal, dental and soft tissues in three types of jaw asymmetry based on our previously reported classification system. The cone-beam computed tomography (CBCT) images of 70 Class III patients were analysed. Group 1 patients showed large shift of menton and synchronous but smaller ramus deviation. The maxillomandibular complex had roll and yaw rotations to the menton-deviation side. Maxillary and dental asymmetry was obvious in transverse and vertical dimensions. Cant of occlusal plane and lip line was apparent. Group 2 patients also exhibited menton and ramus deviation to the same side but the discrepancy in ramus width was larger than menton shift. Asymmetry in Group 2 resulted from a bodily side shift of the maxillomandibular complex without obvious rotation. Group 3 patients had menton and ramus deviated in opposite directions which seemed secondary to a yaw rotation. Double-jaw surgery is generally required for Groups 1 and 3 while Group 2 patients may be successfully treated by mandibular surgery only provided that arch width discrepancy can be managed by orthodontic measures.

Abnormal methylation of spermatozoa induced by benzo(a)pyrene in rats.

Epigenetic mutations caused by pollutants are possibly linked to many diseases. Benzo(a)pyrene (BaP) is one of the most representative air pollutants and has aroused wide concern because of its strong carcinogenicity. The reproductive toxicity induced by BaP has been identified, but little is known about the characteristics of the methylation changes induced by BaP. In this study, a methylated DNA immunoprecipitation sequencing method was used to detect the methylation of sperm DNA of rats exposed to BaP. Compared with the respective genes in normal rats, there were 3227 hypomethylated genes and 828 hypermethylated genes after BaP exposure. Gene ontology enrichment analysis reported that differentially methylated genes (DMGs) were enriched in the localization, single-multicellular organism process and plasma membrane. Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that the DMGs were significantly enriched in the Ras signalling pathway, Rap1 signalling pathway, pancreatic secretion and neuroactive ligand-receptor interaction. DisGeNET disease spectrum analysis showed that DMGs were associated with infertility and certain genetic diseases. Further research needs to be done to explore whether these abnormal methylation are transgenerational.

Antidepressant prescription patterns and CNS polypharmacy with antidepressants among children, adolescents, and young adults: a population-based study in Sweden.

This study examines trends in antidepressant drug dispensations among young people aged 0-24 years in Sweden during the period 2006-2013, as well as prescription patterns and central nervous system (CNS) polypharmacy with antidepressants. Using linkage of Swedish national registers, we identified all Swedish residents aged 0-24 years that collected at least one antidepressant prescription (here defined as antidepressant users) between 1 January 2006 and 31 December 2013 (n = 174,237), and categorized them as children (0-11 years), adolescents (12-17 years), and young adults (18-24 years). Prevalence of antidepressant dispensation rose from 1.4 to 2.1% between 2006 and 2013, with the greatest relative increase in adolescents [by 97.8% in males (from 0.6 to 1.3%) and by 86.3% in females (from 1.1 to 2.1%)]. Most individuals across age categories were prescribed selective serotonin reuptake inhibitors, received their prescriptions from psychiatric specialist care, and had treatment periods of over 12 months. Prevalence of CNS polypharmacy (dispensation of other CNS drug classes in addition to antidepressants) increased across age categories, with an overall increase in prevalence from 52.4% in 2006 to 62.1% in 2013. Children experienced the largest increase in polypharmacy of three or more psychotropic drug classes (4.4-10.1%). Anxiolytics, hypnotics, and sedatives comprised the most common additional CNS drug class among persons who were prescribed antidepressants. These findings show that the dispensation of antidepressants among the young is prevalent and growing in Sweden. The substantial degree of CNS polypharmacy in young patients receiving antidepressants requires careful monitoring and further research into potential benefits and harms.

[Postoperative complications and survival analysis of 1 118 cases of open splenectomy and azygoportal disconnection in the treatment of portal hypertension].

Objective: To analyze the recent postoperative and long-term postoperative complications of open-splenectomy and disconnection in patients with portal hypertension. Methods: There were 1 118 cases with portal hypertension who underwent open splenectomy and azygoportal disconnection from April 2010 to September 2015 at Department of Surgery, People's Liberation Army 302 Hospital. Retrospective case investigation and telephone follow-up were conducted in October 2016. All patients had history of upper gastrointestinal bleeding before operation. Short-term complications after surgery were recorded including secondary laparotomy of postoperative abdominal hemostasis, severe infection, intake disorders, liver insufficiency, postoperative portal vein thrombosis and perioperative mortality. Long-term data including postoperative upper gastrointestinal rebleeding, postoperative survival rate and incidence of postoperative malignancy were recorded, too. GraphPad Prism 5 software for data survival analysis and charting. Results: Postoperative short-term complications in 1 118 patients included secondary laparotomy of postoperative abdominal hemostasis(1.8%, 21/1 118), severe infection(2.9%, 32/1 118), intake disorders(1.0%, 11/1 118), liver dysfunction (1.6%, 18/1 118), postoperative portal vein thrombosis(47.1%, 526/1 118)and perioperative mortality(0.5%, 5/1 118). After phone call following-up, 942 patients' long-term data were completed including 1, 3, 5 years postoperative upper gastrointestinal rebleeding rate(4.4%, 12.1%, 17.2%), 1, 3, 5-year postoperative survival rate(97.0%, 93.5%, 90.3%); the incidence of postoperative malignant tumors in 1, 3 and 5 years were 1.7%, 4.4% and 6.2%. Conclusions: Reasonable choosing of surgical indications and timing, proper performing the surgery process, effective conducting perioperative management of portal hypertension are directly related to the patient's short-term prognosis after portal hypertension. Surgical intervention can reduce the rates of patients with upper gastrointestinal rebleeding, improve survival, and do not increase the incidence of malignant tumors.

To evaluate the correlation between the change of immune system function before and after the treatment of malignant obstructive type jaundice treated with biliary stent.

OBJECTIVE: To evaluate the correlation between the change of immune system function before and after the treatment of malignant obstructive type jaundice (MOJ) treated with a biliary stent. PATIENTS AND METHODS: 148 patients who were admitted to the Department of Digestive System for malignant obstructive jaundice were selected according to the standardized criterion. Amongst the total sample size, 78 were male patients and 70 were female patients, with an average age of (43.6 ± 5.5) years. After admission, the patients completed the blood routine examination and received biliary stent treatment to relieve the sign and symptoms of jaundice. Follow-up observation included total white blood cells, CD4+T cell count, CD8+T cell count, the ratio of CD4+/CD8, neutrophil counts neutrophils percentage, total bilirubin, free bilirubin, alanine aminotransferase (ALT), and inflammatory factors. RESULTS: After three weeks of follow-up visit, CD4+T lymphocyte absolute value of patients markedly increased compared with that of pre-operation, and the difference had statistical significance (p < 0.05). The total bilirubin, free bilirubin, ALT, and inflammatory factors, such as hs-CRP, TNF-α in plasma of patients was significantly lower than that before the operation and the difference was statistically significant (p < 0.05). After six weeks of follow-up visit, the ratio of CD4+/CD8+ increased and the difference had statistical significance (p < 0.05) compared with that before biliary stent implantation. However, the white blood cell and neutrophil granulocyte did not improve significantly. It was found that CD4+/CD8+T lymphocyte had relation with the level of hs-CRP. CONCLUSIONS: The patients with the (MOJ) treated with implanted biliary stent revealed relive in the obstruction of the biliary tract, which will further significantly improve the cholestasis. The ratio of CD4+/CD8+T lymphocyte increased, which will improve the immune system function of the patients, decreases the possibility of infection, and improves the overall survival quality.

Short-term oral gavage administration of adenine induces a model of fibrotic kidney disease in rats.

INTRODUCTION: The adenine model of kidney disease typically involves dietary delivery of adenine over several weeks. This model can be variable in its disease progression and can result in significant mortality. In the current study, the amount of adenine delivered to rats was controlled by utilizing oral gavage administration over a short period in an attempt to induce robust renal pathology while addressing variability and viability of the animals. METHODS: Adenine (150 or 200 mg/kg) was administered via oral gavage for 10 consecutive days, and assessed over a total of 20 days. RESULTS: Both adenine dose groups manifested pathophysiological features of kidney disease such as proteinuria, elevated serum creatinine and BUN, and tubulointerstitial fibrosis. The animals also displayed a decline in glomerular filtration rate. Renal mRNA expression of genes associated with injury, inflammation, and fibrosis (i.e., Col1a1, Acta2, Serpine1, Timp1, Fn-Eda, Tgfb1, Ccl2, Nlrp3, Aqp1 and Ccnd1) were elevated as were urinary biomarkers that have translational utility (i.e., clusterin, KIM-1, MCP-1, OPN, NGAL, B2M, calbindin, and cystatin C). All disease endpoints were more pronounced in the 200 mg/kg group, however, while measures of tissue fibrosis were sustained, there was partial recovery by day 20 in functional readouts. No mortality was observed in either dose group. DISCUSSION: Short-term delivery of adenine via precise gavage delivery induced a robust model with hallmarks of fibrotic kidney disease, had limited variance between animals, and no animal morbidity within the 20 days studied. This model represents a methodical alternative to long-term dietary dosing of adenine.

Effect of CuSO4 and nano copper on serum antioxidant capacity in Weaned piglets.

Copper is the most essential trace element in the animal body. Nano-sized copper particles have been widely used in a number of different animal species in modern medicinal practice. The present study was designed to examine the effect of dietary copper sulfate/(CuSO4) and nano copper supplementation on serum antioxidant capacity of weaning piglets. A total of 28 Duroc piglets (21 days, and weighing ~7 kg) were randomly divided into three equal groups. The control group (n=4) was administered with a normal standard diet; however the CuSO4 (n=12) and nano copper (n=12) groups were treated with 50, 100 and 200 mg/kg/day body weight, respectively. After 28 days, blood serum copper-zinc superoxide dismutase (CuZn-SOD), ceruloplasmin (CP), malondialdehyde (MDA), myeloperoxidase (MPO), total antioxidant capacity (T-AOC), peroxidase (POD), nitric oxide (NO), nitric oxide synthase (NOS), hydrogen peroxide (H2O2) and inhibition of hydroxyl radical (CIHR) were analyzed from all groups. The results indicated that nano copper supplementation has significant (P less than 0.05) effect on the serum antioxidant capability as compared to dietary CuSO4 group in weaned piglets. Nano-size copper 100 mg/kg/day supplementation was confirmed to improve the immunity level by strengthening the antioxidant capacity of weaning piglets. Dietary supplementation with 100 mg/kg body weight nano copper could be a potential substitute for weaned piglets.

Cardiopulmonary resuscitation-from the patient's perspective.

With increasing emphasis on patient autonomy, patients are encouraged to be more involved in end-of-life issues, including the use of extraordinary efforts to prolong their lives. Being able to make anticipatory decisions is seen to promote autonomy, empower patients and optimise patient care. To facilitate shared decision-making, patients need to have a clear and accurate understanding of cardiopulmonary resuscitation (CPR). This study aims to understand the knowledge and perspectives of the local community regarding resuscitation options and end-of-life decision-making and to explore ways to improve the quality of end-of-life discussions. An interviewer-administered survey was conducted with a prospectively recruited group of surgical patients admitted postoperatively to the day surgery ward of a single tertiary institution in Singapore from April to May 2015. The survey, modelled after two validated questionnaires, measured patients' knowledge, attitudes and preferences regarding CPR in a series of 18 questions. Fifty-one out of 67 (76.1%) patients completed the survey. Results indicated that 80.4% (n=41) of participants correctly understood the purpose of CPR, but 64.7% (n=33) did not know of any possible complications of CPR. Less than half (n=21, 41.2%) of participants had thought about life support measures they wanted for themselves. Most of the participants agreed that they should personally be involved in making end-of-life decisions (n=44, 86.3%). Many patients had a poor knowledge of CPR and other resuscitation measures and the majority overestimated the success rate of CPR. However, a majority were receptive to improving their knowledge and keen to discuss end-of-life issues with physicians.

CHIP/Stub1 regulates the Warburg effect by promoting degradation of PKM2 in ovarian carcinoma.

Tumor cells preferentially adopt aerobic glycolysis for their energy supply, a phenomenon known as the Warburg effect. It remains a matter of debate as to how the Warburg effect is regulated during tumor progression. Here, we show that CHIP (carboxyl terminus of Hsc70-interacting protein), a U-box E3 ligase, suppresses tumor progression in ovarian carcinomas by inhibiting aerobic glycolysis. While CHIP is downregulated in ovarian carcinoma, induced expression of CHIP results in significant inhibition of the tumor growth examined by in vitro and in vivo experiments. Reciprocally, depletion of CHIP leads to promotion of tumor growth. By a SiLAD proteomics analysis, we identified pyruvate kinase isoenzyme M2 (PKM2), a critical regulator of glycolysis in tumors, as a target that CHIP mediated for degradation. Accordingly, we show that CHIP regulates PKM2 protein stability and thereafter the energy metabolic processes. Depletion or knockout of CHIP increased the glycolytic products in both tumor and mouse embryonic fibroblast cells. Simultaneously, we observed that CHIP expression inversely correlated with PKM2 levels in human ovarian carcinomas. This study reveals a mechanism that the Warburg effect is regulated by CHIP through its function as an E3 ligase, which mediates the degradation of PKM2 during tumor progression. Our findings shed new light into understanding of ovarian carcinomas and may provide a new therapeutic strategy for ovarian cancer.

Molecular cloning, expression and functional characterization of tumor necrosis factor (TNF) receptor-associated factor (TRAF)-interacting protein (TRIP) in grass carp, Ctenopharyngodon idella.

TRIP (Tumor Necrosis Factor (TNF) Receptor-Associated Factor (TRAF)-Interacting Protein), a member of the TNF superfamily, plays a crucial role in the modulation of inflammation in vertebrates. However, no information about TRIP is available in teleosts. In this study, the full-length cDNA of TRIP, containing a 5'UTR of 112 bp, an ORF of 1359 bp, and a 3'UTR of 29 bp before the poly (A) tail, was cloned from grass carp, Ctenopharyngodon idella. The TRIP gene encoded a protein of 452 amino acids with an estimated molecular mass of 51.06 KD and a predicted theoretical isoelectric point (pI) of 9.11. Quantitative real-time PCR analysis revealed that TRIP mRNA was expressed in all the tissues examined in grass carp, with the highest expression in the kidney, followed by the intestine and thymus. However, lower levels of expression were also detected in fat, spleen, liver, gonad and heart. Subcellular localization and two-hybrid analysis revealed that TRIP was located in the nucleus and that it interacted with TRAF1 and TRAF2 in HEK293T cells. Furthermore, similar to TNF-α, IL-10 and TRIP mRNA expression was upregulated in the spleen of fish fed high-fat or high-carbohydrate diets, suggesting that TRIP might be associated with the response to excessive energy intake. The mRNA relative expression of TRIP was significantly reduced (P < 0.05) after hepatocyte of C. idella was treated with 2 µg/mL lipopolysaccharide (LPS) for 4 h, while the expression levels of inflammatory cytokines TNF-α and IL-10 were significantly increased (P < 0.05). Taken together, these results indicate that TRIP might play important roles in immune defense and has the potential to be used as a anti-inflammation target in grass carp.

A-306989, an inhibitor of adenosine kinase, is renoprotective in rodent models of podocyte, basement membrane, and obstructive injury.

Adenosine (ADO) is an important regulatory purine nucleoside that accumulates at sites of inflammation and tissue injury including in diseases associated with renal pathology. Endogenous levels of ADO may be increased by inhibiting the ADO-metabolizing enzyme, ADO kinase (AK). AK inhibitors have demonstrated protection in rodent models of diabetic nephropathy. To further investigate AK inhibition as a potential mechanism for renal protection, A-306989, a potent non-nucleoside AK inhibitor, was examined in both in vitro and in vivo assays of renal injury. A-306989 prevented podocyte damage (disruption of actin cytoskeleton) and increased podocyte survival following puromycin aminonucleoside (PAN) application in both mouse and human conditionally immortalized podocytes. Prophylactic oral administration of A-306989 (1.5, 5 and 15mg/kg) reduced proteinuria in a dose-dependent manner and repressed pro-inflammatory/fibrotic gene up-regulation; A-306989 was also efficacious when administered two days following the PAN-insult. A-306989 (10 and 30mg/kg) also significantly reduced proteinuria and macrophage infiltration in a rat model of glomerulonephritis. Finally, A-306989 (15 and 50mg/kg) reduced the expression levels of pro-inflammatory/fibrotic genes, and reduced macrophage infiltration (50mg/kg), but did not affect the deposition of interstitial collagen in fibrotic kidneys from mice with unilateral ureter obstruction. A-306989 also had beneficial actions on "quality of life" measures including improving body weight loss. Thus, these data indicate that enhancement of endogenous ADO levels by A-306989 can positively modulate renal pathology and mimic some of the previously reported beneficial actions of ADO A2A receptor agonists.