[Expression of CD7 and its correlation with prognosis in patients with NK/T-cell lymphoma].

Objective: To analysis the expression of CD7 in NK/T-cell lymphoma as well as study the correlations between CD7 and clinical survival and prognosis. Methods: The clinical and pathological indicators of 112 NKTCL patients who were admitted to or consulted at the First Affiliated Hospital of Zhengzhou University between May 2008 and December 2019 were analyzed retrospectively. The CD7 expression in the tumor tissues was detected using immunohistochemistry staining, and the influence of CD7 expression on the survival and prognosis in the patients was analyzed. Results: The CD7 expression rate was 84.82% in 112 NKTCL patients, and its expression was not influenced by sex, age, and the primary site. An analysis of the complete clinical data of 72 patients showed that the CD7 expression was significantly correlated with the PINK score, tumor metastasis, and peripheral blood EBV-DNA level. However, the Ann Arbor stage, bone marrow involvement, B symptoms, IPI/aaIPI score, Ki-67, EBER, TIA-1, Granzyme B, LDH, and ß(2)-MG were not associated with the CD7 expression. The 1-year, 3-year, and 5-year overall survival (OS) rates of the 72 patients were 81.2%, 61.8%, and 58.8%, respectively, and the progression-free survival (PFS) rates were 53.5%, 29.4%, and 24.0%, respectively. The median overall survival (median-OS, mOS) was 81 mon, and the median progression-free survival (median-PFS, mPFS) was 14 mon. The 3-year OS rates in the CD7-positive group and the CD7-negative group were 58.1% and 83.9%, respectively, (P>0.05) . The 3-year PFS rates were 21.7% and 51.9%, respectively (P<0.05) . The univariate analysis showed that age, primary tumor site, Ann Arbor stage, IPI/aaIPI score, PINK score, LDH, ß(2)-microglobulin, EBV-DNA, Ki-67, and CD7 influenced patient prognosis. The multivariate analysis showed that Ann Arbor stage and CD7 were independent prognostic factors for PFS, while PINK score and Ki-67 were independent prognostic factors for OS. Conclusions: The expression rate of CD7 in NKTCL was high and was closely related to poor patient prognosis. The patients with high levels of EBV-DNA, metastatic disease, or high PINK score were more likely to express CD7.

Clinical features and survival impact of EBV-positive diffuse large B-Cell lymphoma with different age cutoffs.

OBJECTIVE: In 2016 WHO classification, EBV +DLBCL of the elderly was replaced by EBV+ DLBCL NOS. This is due to the fact that many young patients of EBV+ DLBCL were found in recent years. PATIENTS AND METHODS: In this study, we retrospectively analyzed clinical features and survival outcomes of EBV positive DLBCL patients in different age groups. All the patients treated at a single center. RESULTS: When we use different ages (40, 50 and 60 years old) as cutoffs, the prevalence of EBV positive DLBCL was 12.0%, 12.3% and 13.0% in younger patients and 19.0%, 15.4% and 13.8% in elder patients respectively. Whatever the age cutoff was, EBV positive associated with unfavorable clinical prognosis in elder groups. When we use 40 and 50 years old as age cutoffs, poor impacts of EBV positive on overall survival and progression-free survival were observed only in elder patients, but not in younger patients. It should be noted that when we use 60 years old as age cutoff, the results were the opposite. CONCLUSIONS: EBV+ DLBCL patients with age of 40 to 60 years old showed poorer prognostic features than EBV- DLBCL patients; however, patients in other age groups did not show evident differences in prognosis between EBV+ DLBCL patients and EBV- DLBCL patients. This finding was not reported before.

Characterization and virus susceptibility of a continuous cell line derived from the brain of Aequidens rivulatus (Günther).

Cell cultures derived from the brain tissues of Aequidens rivulatus (Günther) have been characterized previously. In this study, a continuous cell line ARB8 was further established, and its growth characteristics, transcription and susceptibility to fish viruses-including chum salmon reovirus (CSV), marbled eel infectious pancreative necrosis virus (MEIPNV), grouper nervous necrosis virus (GNNV), giant seaperch iridovirus (GSIV), red seabream iridovirus (RSIV), koi herpesvirus (KHV), herpesvirus anguilla (HVA) and marbled eel polyoma-like virus (MEPyV)-were examined. ARB8 cells that showed epithelioid morphology and were passaged >80 times grew well at temperatures ranging from 25°C to 30°C in L-15 medium containing 5%-15% foetal bovine serum. The cells constitutively transcribed connexion 43, glutamine synthetase, nestin and nkx6-2, which are markers for neural progenitor cells. The cells were highly susceptible to CSV, MEIPNV, GSIV and RSIV and showed the typical cytopathic effect (CPE). However, the cells were resistant to GNNV, KHV, HVA and MEPyV because no significant CPE was noted after infection. Optimal temperatures for virus production ranged from 25°C to 30°C. The results revealed that the neural progenitor cell line ARB8 can potentially serve as a useful tool for investigating fish viruses and isolating new viruses in ornamental cichlid fishes.

Immunocytochemical characterisation of neural stem-progenitor cells from green terror cichlid Aequidens rivulatus.

In this study, cultures of neural stem-progenitor cells (NSPC) from the brain of green terror cichlid Aequidens rivulatus were established and various NSPCs were demonstrated using immunocytochemistry. All of the NSPCs expressed brain lipid-binding protein, dopamine- and cAMP-regulated neuronal phosphoprotein 32 (DARPP-32), oligodendrocyte transcription factor 2, paired box 6 and sex determining region Y-box 2. The intensity and localisation of these proteins, however, varied among the different NSPCs. Despite being intermediate cells, NSPCs can be divided into radial glial cells, oligodendrocyte progenitor cells (OPC) and neuroblasts by expressing the astrocyte marker glial fibrillary acidic protein (GFAP), OPC marker A2B5 and neuronal markers, including acetyl-tubulin, ßIII-tubulin, microtubule-associated protein 2 and neurofilament protein. Nevertheless, astrocytes were polymorphic and were the most dominant cells in the NSPC cultures. By using Matrigel, radial glia exhibiting a long GFAP+ or DARPP-32+ fibre and neurons exhibiting a significant acetyl-tubulin+ process were obtained. The results confirmed that NSPCs obtained from A. rivulatus brains can proliferate and differentiate into neurons in vitro. Clonal culture can be useful for further studying the distinct NSPCs.

Isolation of a novel polyomavirus, related to Japanese eel endothelial cell-infecting virus, from marbled eels, Anguilla marmorata (Quoy & Gaimard).

Marbled eels, Anguilla marmorata (Quoy & Gaimard), cultured in Taiwan exhibited haemorrhage and mortality in January 2012. The severely diseased eels bled from the gills and showed congestion of the central venous sinus of the gill filaments and haemorrhage throughout the body similar to viral endothelial cell necrosis of eel. In this study, a novel polyomavirus (AmPyV) was isolated from the diseased eels using the AMPF cell line established from the pectoral fin of healthy marbled eels. AmPyV was found to encode a long T-antigen orthologous gene. Phylogenetic analysis showed that AmPyV was closely related to Japanese eel endothelial cell-infecting virus. PCR assays revealed AmPyV infection throughout the systemic organs. AmPyV proliferated in the AMPF, EK-1 and EO-2 cells at temperatures 25-30 °C, and the progeny virus yields were 10(7.0) , 10(7.4) and 10(7.7) TCID50  mL(-1) , respectively. The purified virions were icosahedral particles, 70-80 nm in diameter. No clinical signs or mortality was observed among the eels injected with the virus; however, the virus was reisolated from the brain, eyes, kidneys, fins and gills of infected eels 2 month after injection. Our results suggest that AmPyV exhibits a latent infection. Pathogen of the disease needs to study further.

Effect of biological behavior and clinical significance of maspin gene on cervical squamous carcinoma SiHa cell.

OBJECTIVE: This study was performed to evaluate the effect of mammary serine protease inhibitor (maspin) overexpression on human cervical squamous carcinoma (SCC) SiHa cell proliferation and apoptosis in vitro. MATERIAL AND METHODS: Recombinant plasmid pcDNA3-maspin was stably transfected into human cervical SCC SiHa cell. Maspin mRNA was determined by RT-PCR, whereas maspin protein was detected by Western blot analysis and immunocytochemistry (IHC). Cell proliferation activity was measured by MTT method. Apoptosis rate and cell cycle distribution were detected by flow cytometry to understand the changes in the cell biological characteristics. RESULTS: The strengthened expression of the maspin gene in the SiHa cell was confirmed by RT-PCR, Western blot, and immunocytochemistry (IHC) (p < 0.05). Suppressed proliferation activity and increased apoptosis rate of SiHa-m (maspin stable transfected) versus SiHa and SiHa-vector cell (SiHa-pc3) were shown by MTT and flow cytometry (p < 0.05). SiHa and SiHa-pc3-had no statistical significance (p > 0.05). CONCLUSION: The results showed that maspin gene can significantly inhibit human cervical SCC SiHa cell proliferation and effectively slow cancer growth. Maspin may be a new molecular target in the gene therapy of human cervical SCC.

A novel chemical uncoupler ameliorates obesity and related phenotypes in mice with diet-induced obesity by modulating energy expenditure and food intake.

AIMS/HYPOTHESIS: Decreasing mitochondrial coupling efficiency has been shown to be an effective therapy for obesity and related metabolic symptoms. Here we identified a novel mitochondrial uncoupler that promoted uncoupled respiration in a cell type-specific manner and investigated its effects on modulation of energy metabolism in vivo and in vitro. METHODS: We screened a collection of mitochondrial membrane potential depolarising compounds for a novel chemical uncoupler on isolated skeletal muscle mitochondria using a channel oxygen system. The effect on respiration of metabolic cells (L6 myotubes, 3T3-L1 adipocytes and rat primary hepatocytes) was examined and metabolic pathways sensitive to cellular ATP content were also evaluated. The chronic metabolic effects were investigated in high-fat diet-induced obese mice and standard diet-fed (SD) lean mice. RESULTS: The novel uncoupler, CZ5, promoted uncoupled respiration in a cell type-specific manner. It stimulated fuel oxidation in L6 myotubes and reduced lipid accumulation in 3T3-L1 adipocytes but did not affect gluconeogenesis or the triacylglycerol content in hepatocytes. The administration of CZ5 to SD mice increased energy expenditure (EE) but did not affect body weight or adiposity. Chronic studies in mice on high-fat diet showed that CZ5 reduced body weight and improved glucose and lipid metabolism via both increased EE and suppressed energy intake. The reduced adiposity was associated with the restoration of expression of key metabolic genes in visceral adipose tissue. CONCLUSIONS/INTERPRETATION: This work demonstrates that a cell type-specific mitochondrial chemical uncoupler may have therapeutic potential for treating high-fat diet-induced metabolic diseases.

Effects of temperature and trehalose on foam separation of nisin from the culture broth produced by Lactococcus lactis subspecies lactis W28.

Nisin is an antimicrobial peptide, an important biopreservative, and it is produced by certain strains of Lactococcus lactis ssp. lactis. In this paper, a foam separation technique was used for the separation of nisin from its culture broth, and the effects of temperature and trehalose on the performance of foam separation of nisin were studied to increase the enrichment ratio and recovery percentage of nisin and decrease the inactivity percentage of nisin. The results showed that temperature and trehalose significantly affected the performance of foam separation of nisin. Under the optimum conditions of 50°C temperature, 150-mL/min air flow rate, 400-mL initial loading liquid volume, and 1-g/L trehalose concentration, the maximum enrichment ratio, recovery percentage, and the minimum inactivity percentage of nisin reached 23.7, 84.1%, and 5.9%, respectively, which were, respectively, 5.04, 0.93, and 1.03 times more than those under the conditions of 20°C temperature, 150-mL/min air flow rate, 400-mL initial loading liquid volume, and no trehalose addition. These results indicated that the change of temperature and the addition of trehalose could improve the performance of foam separation of nisin.

Safety assessment of mushroom ß-glucan: subchronic toxicity in rodents and mutagenicity studies.

Mushroom ß-glucan, a polymer of ß-(1,3/1,6)-glucan, has been claimed for its health benefits. The objective of this study was to assess the safety in-use of mushroom ß-glucan as dietary supplement and food ingredient. Hence, a subchronic toxicity and mutagenicity studies were conducted. In the subchronic toxicity study, Sprague Dawley rats (12/sex/group) were administered (gavage) mushroom ß-glucan at dose levels of 0, 500, 1000 and 2000 mg/kg body weight (bw)/day for 90 days. As compared to control group, administration of ß-glucan did not result in any toxicologically significant treatment-related changes in clinical observations, ophthalmic examinations, body weights, body weight gains, feed consumption, and organ weights. No adverse effects of the ß-glucan on the hematology, serum chemistry parameters, urinalysis or terminal necropsy (gross or histopathology findings) were noted. The results of mutagenicity studies as evaluated by gene mutations in Salmonella typhimurium, in vitro chromosome aberrations and in vivo micronucleus test in mouse did not reveal any genotoxicity of ß-glucan. Based on the subchronic study, the no observed-adverse-effect level (NOAEL) for mushroom ß-glucan was determined as 2000 mg/kgbw/day, the highest dose tested.

Evaluation of spatial processing in virtual reality using functional magnetic resonance imaging (FMRI).

While the ecological validity of virtual reality (VR) applications is usually assessed by behavioral data or interrogation, an alternative approach on a neuronal level is offered by brain imaging methods. Because it is yet unclear if 3D space in virtual environments is processed analogically to the real world, we conducted a study investigating virtual spatial processing in the brain using functional magnetic resonance imaging (fMRI). Results show differences in VR spatial brain processing as compared to known brain activations in reality. Identifying differences and commonalities of brain processing in VR reveals limitations and holds important implications for VR therapy and training tools. When VR therapy aims at the rehabilitation of brain function and activity, differences in brain processing have to be taken into account for designing effective VR training tools. Furthermore, for an evaluation of possible restoration effects caused by VR training, it is necessary to integrate information about the brain activation networks elicited by the training. The present study provides an example for demonstrating the benefit of fMRI as an evaluation tool for the mental processes involved in virtual environments.

Serum uric acid is associated with metabolic risk factors for cardiovascular disease in the Uygur population.

The prevalence of hyperuricemia is low in Uygurs, who have a high prevalence of cardiovascular risk factors such as hypertension, overweight-obesity, dyslipidemia, hyperglycemia, and insulin resistance (IR). This study sought to investigate the relationships between serum uric acid (UA) and these risk factors in this population. A cross-sectional study was conducted in Uygurs (859 males, 1268 females) aged 20 to 70 years. Demographic data, systolic blood pressure (SBP), diastolic blood pressure (DBP), body mass index (BMI), and fasting and postprandial blood were obtained, and biological measurements were determined. The mean of BMI, SBP, DBP, total cholesterol, high-density lipoprotein cholesterol (HDL-c), low-density lipoprotein cholesterol (LDL-c), triglycerides, fasting blood glucose, fasting insulin, and homeostasis model assessment insulin resistance index (HOMA-IR), and the prevalence of hypertension, IR, hyperglycemia, overweight-obesity, hypercholesteremia, hyper-LDL-c, and hypertriglyceridemia increased with UA but the prevalence of hypo-HDL-c decreased (p < 0.05). Logistic regression analysis showed that the odds ratios for IR, overweight-obesity, hypercholesteremia, hyper-LDL-c, and hypertriglyceridemia against the lowest UA group increased but decreased for hypo-HDL-c (p < 0.05). The UA in the hypo-HDL-c group was lower than that of the controls; the prevalence of hypo-HDL-c in hyperuricemia subjects was lower than in those with normal UA (p < 0.05). But the opposite results were observed between overweight-obesity, hyperglycemia, IR, hypercholesteremia, hypertriglyceridemia, and hyper-LDL-c and correspondence controls, respectively (p < 0.05). In Uygur, elevated UA is associated with overweight-obesity, hypercholesteremia, hyper-LDL-c, hypertriglyceridemia, hyperglycemia, and IR. The HDL-c level significantly increases with UA, whereas the prevalence of hypo-HDL-c decreases. Further studies are needed to clarify why UA is positively correlated to HDL-c.

Molecular cloning and gene expression of the gonadotropin-releasing hormone receptor in the orange-spotted grouper, Epinephelus coioides.

The objective of this study was to investigate the molecular mechanisms of gonadotropin-releasing hormone receptor (GnRH-R) involved in the endocrine regulation of reproduction in the orange-spotted grouper, Epinephelus coioides. The full-length cDNA encoding GnRH-R type I was successfully cloned from the pituitary by reverse-transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA end (RACE) methods in the grouper. The complete GnRH-R type I cDNA is 1607 bp, which includes an open reading frame of 1092 bp encoding a protein of 364 amino acids, a seven-alpha helix transmembrane domain, a N-terminal extracellular domain, and a C-terminal cytoplasmic domain. The expression of GnRH-R type I was found to be highest in the pituitary. An intramuscular injection of various GnRH types in vivo was attempted. The expression of GnRH-R type I was stimulated by a single injection of salmon GnRH, while in the case of chicken GnRH II treatment, the expression of GnRH-R type I was inhibited. This suggests that the action of chick GnRH II is probably enhanced through the GnRH receptor of different forms. Furthermore, none of them were expressed by an injection of seabream GnRH, and this is likely attributed to the injection dose being below the threshold level, and this remains to be further examined. In conclusion, GnRHs of various types are effective in stimulating the expression of gonadotropins through various forms of the GnRH-R, and multiple forms of the receptor gene likely exist in teleosts.

Pharmacokinetics of mycophenolic acid and its glucuronide after a single and multiple oral dose of mycophenolate mofetil in Chinese renal transplantation recipients.

PURPOSES: To compare the pharmacokinetic parameters of mycophenolic acid (MPA) and mycophenolic acid glucuronide (MPAG) after single and multiple oral administration of mycophenolate mofetil (MMF) in Chinese renal transplant patients with those of recipients in other countries. METHODS: Twelve Chinese renal transplant patients after an overnight fast received a single 1-g dose of MMF before renal transplantation. Thereafter the patients received 1 g MMF twice a day up to and on the day 12 after renal transplantation. The concentrations of MPA and MPAG were simultaneously measured by RP-HPLC. The concentration-time data were examined with Drug and Statistics pharmacokinetic software. Using paired samples t test (self-contrast) with SPSS statistical software (alpha = 0.05), we compared the pharmacokinetic parameters between single and multiple doses. RESULTS AND DISCUSSIONS: The MPA concentration-time profiles were fitted to a two-compartment open model; MPAG concentration-time profiles were fitted to a single-compartment open model. Compared with the literature reports, the main pharmacokinetic parameters of MPA and MPAG shown by our research revealed some differences. The parent drug MMF was undetectable in plasma during oral administration. A secondary peak of MPA occurred at 6 to 10 hours, which was attributed to enterohepatic recirculation. There was significant variation in MPA and MPAG plasma concentration-time data among subjects. It is suggested that therapeutic drug monitoring should be applied for dosage optimization.

Therapeutic drug monitoring of mycophenolic acid can be used as predictor of clinical events for kidney transplant recipients treated with mycophenolate mofetil.

OBJECTIVE: This study was designed to investigate the relationship between clinical events and the pharmacokinetics of mycophenolic acid (MPA) in adult renal transplant patients. METHODS: Thirty-seven adult kidney transplant recipients received a cyclosporine, mycophenolate mofetil, and steroids. MPA-AUC(0-12) was obtained before and 12 days after grafting by RP-HPLC. Predose blood samples were measured for MPA-C(0) were gathered from all the recipients at 4, 12, and 21 days as well as 1, 1.5, 2, 2.5, 3, and 6 months after grafting. The clinical events at corresponding time points were recorded to correlate with each MPA-C(0) value. RESULTS: In addition to single-dose and multidose MPA-AUC(0-12), 357 MPA-C(0) values were obtained from 37 patients. The 357 units were divided into three subgroups: group A patients, including 239 units (66.9%), experienced uneventful outcomes; group B of 100 units (28.0%) showed MPA-related side effects, and group C, 18 units (5.0%) of acute rejection episodes. MPA-C(0) for groups A, B, and C were 0.8416 +/- 0.1373 mg/L, 1.5903 +/- 0.3741 mg/L and 0.6057 +/- 0.2338 mg/L, respectively (P < .001 between groups A and B, groups B and C, and P = .021 between groups A and C). The three groups were also divided into an initial phase (< or =1 month, 251 units) and a stable phase (>1 month, 106 units). The relationship between MPA-C(0) and associated clinical events was also investigated. CONCLUSION: Our results suggested a relationship between MPA pharmacokinetics and clinical events. The MPA-C(0) might be an appropriate pharmacokinetic monitoring parameter for kidney transplantation.

O-methyl nakafuran-8 lactone, a new sesquiterpenoid from a hainan marine sponge Dysidea sp.

A new sesquiterpenoid, O-methyl nakafuran-8 lactone (1) has been isolated from a Hainan sponge Dysidea sp. and the structure of the new compound proposed by spectral data, was confirmed by X-ray diffraction analysis. The complete 1H- and 13C-NMR assignments were made on the basis of detailed 2D NMR spectral analysis. Compound 1 showed strong inhibitory bioactivity against PTP1B with IC50 value of 1.58 microM.

Development of a K562 cell-based assay for screening anticancer agents.

AIM: To develop a leukemia cell line K562-based assay for high-throughput screening. METHODS: The screening was carried out on 96-well plates with monitoring cell proliferation by a combined 3-[4,5-dimethylthiazol-2-yl]-5-[3-carboxymethoxyphenyl]-2-[4-sulfophenyl]-2H-tetrazolium (MTS)/phenazine methosulfate (PMS) method. Conditions for evaluating effects on the proliferation of K562 cells by individual compounds on the 96-well plates were optimized. RESULTS: A set of 800 small organic compounds was screened for anticancer activity by this cell-based assay, with consumption of each compound at 500 ng. Eleven compounds were identified with >80 % inhibitory activity at 5 mg/L, among which 9 compounds were confirmed by subsequent testing at multiple concentrations. The most potent compound showed an IC50 at 170 nmol/L, and there were total of 7 compounds showed IC50 less than 10 micromol/L. CONCLUSION: The high-throughput method using K562 cell line is fast, economical, effective, and practical in identifying inhibitors as potential therapeutic agents for cancer.