A Novel Nomogram Model to Predict the Recurrence-Free Survival and Overall Survival of Hepatocellular Carcinoma.

Background: Treatments for patients with early-stage hepatocellular carcinoma (HCC) include liver transplantation (LT), liver resection (LR), radiofrequency ablation (RFA), and microwave ablation (MWA), are critical for their long-term survival. However, a computational model predicting treatment-independent prognosis of patients with HCC, such as overall survival (OS) and recurrence-free survival (RFS), is yet to be developed, to our best knowledge. The goal of this study is to identify prognostic factors associated with OS and RFS in patients with HCC and develop nomograms to predict them, respectively. Methods: We retrospectively retrieved 730 patients with HCC from three hospitals in China and followed them up for 3 and 5 years after invasive treatment. All enrolled patients were randomly divided into the training cohort and the validation cohort with a 7:3 ratio, respectively. Independent prognostic factors associated with OS and RFS were determined by the multivariate Cox regression analysis. Two nomogram prognostic models were built and evaluated by concordance index (C-index), calibration curves, area under the receiver operating characteristics (ROC) curve, time-dependent area under the ROC curve (AUC), the Kaplan-Meier survival curve, and decision curve analyses (DCAs), respectively. Results: Prognostic factors for OS and RFS were identified, and nomograms were successfully built. Calibration discrimination was good for both the OS and RFS nomogram prediction models (C-index: 0.750 and 0.746, respectively). For both nomograms, the AUC demonstrated outstanding predictive performance; the DCA shows that the model has good decision ability; and the calibration curve demonstrated strong predictive power. The nomograms successfully discriminated high-risk and low-risk patients with HCC associated with OS and RFS. Conclusions: We developed nomogram survival prediction models to predict the prognosis of HCC after invasive treatment with acceptable accuracies in both training and independent testing cohorts. The models may have clinical values in guiding the selection of clinical treatment strategies.

Sulforaphane suppresses carcinogenesis of colorectal cancer through the ERK/Nrf2­UDP glucuronosyltransferase 1A metabolic axis activation.

The long pre­cancerous state of colorectal cancer (CRC) provides an opportunity to prevent the occurrence and development of CRC. The detoxification of CRC food­borne carcinogenic heterocyclic amines is highly dependent on UDP glucuronosyltransferase 1A (UGT1A)­mediated glucuronidation. Sulforaphane (SFN), a phytochemical, possesses antioxidant, anti­inflammatory and anticarcinogenic effects on the prevention of CRC. Previous studies revealed that SFN upregulates the expression of UGT1A. The aim of the present study was to investigate the regulatory mechanism of SFN­induced UGT1A upregulation and provide novel understanding on the basic research and chemoprevention of CRC. In the present study, the viability and proliferation of CRC cells (HT­29 and SW480) treated with SFN were assessed by MTT, colony formation and EdU assays. Flow cytometry was used to detect the cell cycle arrest and apoptosis of cells treated with different concentrations of SFN. The motility of cells was determined by wound healing and Transwell assays. Nuclear factor, erythroid 2 like 2 (Nrf2) short hairpin RNA (shRNA) and negative control shRNA lentiviruses were used for cell transfection. Reverse transcription­quantitative polymerase chain reaction and western blotting were employed to verify the role of Nrf2 in SFN­induced UGT1A. HT­29 and SW480 cells were divided into a control, an SFN and a PD98059 [an extracellular signal­regulated kinase (ERK) inhibitor] + SFN group. Western blotting detected the protein levels of Nrf2 and UGT1A. Intracellular levels of reactive oxygen species (ROS) were detected using a reactive oxygen assay kit. The results revealed that SFN inhibits cell proliferation and colony formation, promotes apoptosis, and reduces the migratory ability of CRC cells. The phosphorylation of ERK induced by SFN promoted Nrf2 accumulation. Furthermore, a significant increase in the levels of UGT1A was observed, which coincided with SFN­induced upregulation of Nrf2 levels in nuclear fractions. Pretreatment with PD58059 reversed the SFN­induced subcellular translocation of Nrf2 and the expression of UGT1A. In addition, SFN­induced high levels of ROS in CRC cells may be associated with the ERK signaling pathway. Collectively, these results indicated that SFN inhibited the proliferation of CRC cells and upregulated the expression of UGT1A in CRC cells via the ERK/Nrf2 signaling pathway.

Metformin produces anxiolytic-like effects in rats by facilitating GABAA receptor trafficking to membrane.

BACKGROUND AND PURPOSE: Altered function or expression of GABAA receptors contributes to anxiety disorders. Benzodiazepines are widely prescribed for the treatment of anxiety. However, the long-term use of benzodiazepines increases the risk of developing drug dependence and tolerance. Thus, it is urgent to explore new therapeutic approaches. Metformin is widely used to treat Type 2 diabetes and other metabolic syndromes. However, the role of metformin in psychiatric disorders, especially anxiety, remains largely unknown. EXPERIMENTAL APPROACH: We examined the effects of metformin on anxiety-like behaviour of rats in open field test and elevated plus maze test. We also observed the effect of metformin (10 µM, in vitro; 100 mg·kg-1 , in vivo) on the trafficking of GABAA receptors, as mechanisms underlying the anxiolytic effects of metformin. KEY RESULTS: Metformin (100 mg·kg-1 , i.p. 30 min) displayed a robust and rapid anxiolytic effect, without tolerance. Metformin up-regulated the surface expression of GABAA receptors and increased miniature inhibitory postsynaptic currents (mIPSCs). AMP-activated protein kinase (AMPK) activated by metformin-induced stimulation of forkhead box O3a (FoxO3a) transcriptional activity, followed by increased expression of GABAA receptor-associated protein (GABARAP) and its binding to GABAA receptors finally resulted in the membrane insertion of GABAA receptors. CONCLUSIONS AND IMPLICATIONS: Metformin increased mIPSCs by up-regulating the membrane insertion of GABAA receptors, via a pathway involving AMPK, FoxO3a, and the GABAA receptor-associated protein. Thus metformin has a potential new use in the treatment of anxiety disorders.

Pharmacokinetics and tolerability of oral dosage forms of huperzine a in healthy Chinese male volunteers: a randomized, single dose, three-period, six-sequence crossover study.

Huperzine A is a potent, reversible, and blood-brain barrier permeable acetylcholinesterase inhibitor. The aim of this study was to compare the pharmacokinetics, tolerability, and bioavailability of two formulations with the established reference formulation of huperzine A in a fasting, healthy Chinese male population. This was a randomized, single-dose, 3-period, 6-sequence crossover study. The plasma concentrations of huperzine A were determined by liquid chromatography tandem mass spectrometry. Tolerability was assessed based on subject interview, vital sign monitoring, physical examination, and routine blood and urine tests. The mean (SD) pharmacokinetic parameters of the reference drug were Cmax, 1.550 (0.528) ng/mL; t1/2, 12.092 (1.898) h; AUC0-72h, 17.550 (3.794) ng·h/mL. Those of the test formulation A and test formulation B were Cmax, 1.412 (0.467), 1.521 (0.608) ng/mL; t1/2, 12.073 (2.068), 12.271 (1.678) h; AUC0-72h, 15.286 (3.434) ng·h/mL, 15.673 (3.586) ng·h/mL. The 90% confidence intervals for the AUC0-72h and Cmax were between 0.80 and 1.25. No adverse events were reported by the subjects or found with results of clinical laboratory test. The test and reference products met the regulatory criteria for bioequivalence in these fasting, healthy Chinese male volunteers. All three formulations appeared to be well tolerated.

Pharmacokinetics and Tolerability of Oral Dosage Forms of Huperzine A in Healthy Chinese Male Volunteers: a Randomized,Single Dose, Three-period, Six-sequence Crossover Study / 华中科技大学学报（医学）（英德文版）

Huperzine A is a potent,reversible,and blood-brain barrier permeable acetylcholinesterase irhibitor.The aim of this study was to compare the pharmacokinetics,tolerability,and bioavailability of two formulations with the established reference formulation of huperzine A in a fasting,healthy Chinese male population.This was a randomized,single-dose,3-period,6-sequence crossover study.The plasma concentrations of huperzine A were determined by liquid chromatography tandem mass spectrometry.Tolerability was assessed based on subject interview,vital sign monitoring,physical examination,and routine blood and urine tests.The mean (SD) pharmacokinetic parameters of the reference drug were Cmax,1.550 (0.528) ng/mL;t1/2,12.092 (1.898) h;AUC0-72h,17.550 (3.794) ng.h/mL.Those of the test formulation A and test formulation B were Cmax,1.412 (0.467),1.521 (0.608) ng/mL;t1/2,12.073 (2.068),12.271 (1.678) h;AUC0-72h,15.286 (3.434) ng.h/mL,15.673 (3.586) ng.h/mL.The 90％ confidence intervals for the AUC0-72h and Cmax were between 0.80 and 1.25.No adverse events were reported by the subjects or found with results of clinical laboratory test.The test and reference products met the regulatory criteria for bioequivalence in these fasting,healthy Chinese male volunteers.All three formulations appeared to be well tolerated.

Impairment of Biofilm Formation by TiO2 Photocatalysis through Quorum Quenching.

The release of nanomaterials into the environment, due to their massive production and application today, has caused ecological and health safety concerns. Semiconductor photocatalysts like TiO2 exhibit cytotoxicity to bacterial cells when exposed to UV irradiation. However, information about their impacts on individual or group bacterial behaviors is limited. In this work, the biofilm formation of Escherichia coli K12 in the presence of TiO2 with and without UV irradiation was investigated and biofilm formation was found not to be affected under the sole application of TiO2 or UV irradiation. However, biofilm development was substantially delayed by TiO2 under UV irradiation, although no obvious cytotoxicity to cell growth was observed. The reactive oxygen species photogenerated by TiO2 were found to quench the autoinducer 2 (AI-2) signals secreted by E. coli K12. As a result, the initiation of quorum sensing for biofilm formation activated by AI-2 was restrained. The expressions of two biofilm-formation-related genes, motA and rcsB, were also suppressed. A dose of an AI-2 precursor, 4,5-dihydroxy-2,3-pentanedione, effectively restored the biofilm development. These results show that the photoexcited TiO2 could suppress biofilm formation through quenching AI-2 signals. This work may facilitate a better understanding about the ecological effects of increasingly released nanomaterials and provide implications for development of antifouling membranes.

Age- and sex-based patterns of positional behavior and substrate utilization in the golden snub-nosed monkey (Rhinopithecus roxellana).

Body mass plays an important role in primate positional behavior and in sexually dimorphic arboreal primate species may influence how immature and adult individuals travel through the forest canopy and access food resources. In this study, we examined age- and sex-based patterns of positional behavior and substrate utilization in wild golden snub-nosed monkeys (Rhinopithecus roxellana), an endangered species of Asian colobine. Our results indicated that among all age and sex classes, sitting was the most common feeding and resting posture and during travel, quadrupedal walking was the dominant locomotor behavior. Despite the fact that adult male R. roxellana are reported to exhibit a body mass nearly two times that of adult females, we found no significant sex differences in the positional repertoire during feeding and traveling. In addition, we found that while infants and juveniles used similar postural and locomotor behaviors as their adult counterparts, younger golden snub-nosed monkeys more frequently engaged in risky or escape-oriented behaviors such as climbing, running, leaping, and forelimb suspension. With increasing age, the use of quadrupedal walking and dropping (downward in-air displacement of body mass that does not require hindlimb propulsion) increased and the use of leaping, suspensory postures, and bridging decreased. Finally, given differences in the positional repertoire of adult and immature golden snub-nosed monkeys, we argue that studies of ontogenetic patterns of positional behavior should emphasize what it takes to survive at each life stage rather than what it takes to match an adult repertoire.

Expression level of valosin containing protein is associated with prognosis of primary orbital MALT lymphoma.

OBJECTIVE: To investigate whether the expression level of valosin-containing protein (VCP) is correlated with the prognosis of primary orbital mucosa-associated lymphoid tissue (MALT) lymphoma. METHODS: VCP expression in 58 samples from primary orbital MALT lymphoma patients was determined by immunohistochemisty using monoclonal antibodies. Correlations between VCP expression level and prognosis were clarified by statistical analysis. RESULTS: It was found that the percentage of VCP positive cells in samples of primary orbital MALT lymphoma ranged from 32% to 95%. The samples were divided into two groups (level 1 and level 2) according to the median value (45%) of the percentage of VCP positive cells. It was found that the expression level of VCP was significantly correlated with recurrence (P=0.003) and tumor size (P=0.008). At the same time, the 5-year disease-free and overall survival rate of patients of level 1 was significantly better than that of level 2 (P=0.001; P=0.032). There was no observed correlation between the expression level of VCP and other clinical features. CONCLUSION: VCP could be a useful marker for predicting the prognosis of primary orbital MALT lymphoma.