Author Correction: Correlation between expression levels of lncRNA UCA1 and miR-18a with prognosis of hepatocellular cancer.

Correction to: European Review for Medical and Pharmacological Sciences 2020; 24 (7): 3586-3591-DOI: 10.26355/eurrev_202004_20820-PMID: 32329833, published online on 15 April 2020. After publication, the authors noticed some mistakes in the manuscript and applied to issue the following changes: the legend of Table I, the date of enrollment of the patients in the section titled "Baseline Characteristics of HCC Patients", the date of follow-up reported in the abstract and in the section entitled "Postoperative Follow-Up". The authors also applied to modify the survival curve of Figure 3 due to misuse of data in the statistical analysis. There are amendments to this paper. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/20820.

A targeted metabolomic profiling of plasma acylcarnitines in nonalcoholic fatty liver disease.

OBJECTIVE: Nonalcoholic fatty liver disease (NAFLD) has become a common liver disorder caused by lipid accumulation and insulin resistance (IR). Acylcarnitines have become a new biomarker of IR. However, their roles in NAFLD are still poorly studied. Thus, we performed a targeted metabolomic analysis to study the level of plasma acylcarnitines in patients with NAFLD. MATERIALS AND METHODS: The levels of 34 plasma acylcarnitines were measured by a targeted metabolomic approach in NAFLD patients (n = 50) and in healthy control subjects (n = 50) by liquid chromatography-tandem mass spectrometry. Detailed demographic and clinical characteristics of all subjects were also analyzed. RESULTS: The clinical presentation of IR was identified in the NAFLD group but not in the healthy control group. Significant differences were found in the levels of several short-, medium- and long-chain acylcarnitines. A high degree of correlation (r>0.7) was found between even-numbered-carbon long-chain acylcarnitines in NAFLD patients. The area under the receiver operator characteristic of long-chain acylcarnitines, especially C20 (AUC=0.952), C16:1 (AUC=0.949) and C14:1OH (AUC=0.944) acylcarnitines, was greater in NAFLD patients than in healthy control subjects. CONCLUSIONS: The accumulation and disorders of acylcarnitines are associated with NAFLD. A positive correlation between even-numbered-carbon long-chain acylcarnitines was found, and these even-numbered-carbon long-chain acylcarnitines. could be used as potential novel screening markers for nonalcoholic fatty liver disease.

Correlation between expression levels of lncRNA UCA1 and miR-18a with prognosis of hepatocellular cancer.

OBJECTIVE: To uncover the prognostic potentials of long non-coding RNA (lncRNA) UCA1 and miR-18a in hepatocellular cancer (HCC). PATIENTS AND METHODS: Expression levels of UCA1 and microRNA-18a (miR-18a) in HCC tissues and adjacent normal ones harvested from 55 HCC patients were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Clinical data of HCC patients were recorded, including pathological grading, tumor staging, intrahepatic metastasis, serum level of α-fetoprotein (AFP), tumor size, tumor number, recurrence, etc. Based on the median levels of UCA1 and miR-18a, enrolled HCC patients were classified into high-level and low-level group. Potential correlation between expression levels of UCA1 and miR-18a with survival of HCC patients was analyzed. The 4-year follow-up data of HCC patients were collected for analyzing factors that may influence prognosis in HCC patients by the Cox regression model. RESULTS: UCA1 was upregulated and miR-18a was downregulated in HCC tissues. HCC patients with stage III-IV, tumor size ≥5 cm or multiple tumors expressed high level of UCA1. Besides, HCC patients with stage I-II, non-intrahepatic metastasis or primarily diagnosed expressed a relatively low level of miR-18a. High-level UCA1 and low-level miR-18a predicted worse prognosis in HCC patients. Cox regression analysis revealed that tumor node metastasis (TNM) staging, intrahepatic metastases, postoperative recurrences, and UCA1 were risk factors for HCC, while miR-18a was the protective factor. CONCLUSIONS: LncRNA UCA1 is upregulated and miR-18a is downregulated in HCC tissues. High-level UCA1 and low-level miR-18a are closely linked to poor prognosis in HCC.

Proteobacteria community compositions correlate with bronchiectasis severity.

BACKGROUND: Proteobacteria contributes to airway inflammation and poor clinical outcomes in bronchiectasis. OBJECTIVE: To compare sputum Proteobacteria compositions according to bronchiectasis severity. METHODS: Sputum samples collected from 106 patients with stable bronchiectasis and 17 healthy subjects were split for 16srRNA sequencing and biomarker measurement. Pairwise changes in Proteobacteria compositions among 22 of 106 patients during stability, exacerbations and convalescence were compared. Patients were stratified based on the Bronchiectasis Severity Index (BSI). RESULTS: Respectively 44, 34 and 28 patients had mild, moderate and severe bronchiectasis. A higher BSI was associated with a greater relative abundance of Proteobacteria and lower Shannon-Wiener diversity index, Simpson diversity index and bacterial richness. Similar findings applied at genera levels. Proteobacteria and Pseudomonas were the major phylum and genus, respectively, contributing to community similarity in moderate-to-severe bronchiectasis. These significant correlations were not observed in those in whom Pseudomonas aeruginosa was not isolated. Proteobacteria abundance correlated with lung function, but not sputum inflammatory biomarkers in severe bronchiectasis. Proteobacteria compositions in severe bronchiectasis were less likely to change significantly during exacerbations and convalescence. CONCLUSION: Proteobacteria compositions (particularly culturable Pseudomonas abundance) were correlated with bronchiectasis severity. Proteobacteria and Pseudomonas contributed most to community similarity in patients with a higher BSI, indicating microbial targets for interventions in severe bronchiectasis.

Correlations of SOX9 expression with serum IGF1 and inflammatory cytokines IL-1α and IL-6 in skin lesions of patients with acne.

OBJECTIVE: To study the correlations of sex determining region Y-box 9 (SOX9) expression with serum type-1 insulin-like growth factor (IGF-1), interleukin-1α (IL-1α), and interleukin-6 (IL-6) in skin lesion tissues of patients with acne. PATIENTS AND METHODS: Six patients with acne who were treated for the first time in our outpatient clinic from June 2017 to July 2017 were selected as observation group, and 6 normal subjects were selected as control group. The expression of SOX9 was detected by immunohistochemistry. The protein expressions of IGF-1, IL-1α, and IL-6 were detected by enzyme-linked immunosorbent assay (ELISA). SOX9 was detected by quantitative polymerase chain reaction (qPCR). RESULTS: Compared with that in control group, the expression of SOX9 in observation group was significantly increased (p < 0.05). Compared with those in control group, the expressions of IGF-1, IL-1α and IL-6 in observation group were significantly increased (p < 0.05). Compared with that in control group, the mRNA expression of SOX9 in observation group was significantly increased (p < 0.05). SOX9 was positively correlated with IGF-1, IL-1α and IL-6. CONCLUSIONS: The expressions of SOX9, IGF-1, IL-1α, and IL-6 in skin lesion tissues of patients with acne are increased, and SOX9 is positively correlated with IGF-1, IL-1α, and IL-6 and can be used as a target for the treatment of acne inflammation.

Plasma melatonin levels do not differ in SLE patients.

OBJECTIVE: This study aims to investigate the plasma melatonin levels in systemic lupus erythematosus (SLE) patients and its relationship with clinical and laboratory features. PATIENTS AND METHODS: A total of 90 patients with SLE (82 females, 8 males; mean age 37.86 ± 13.98 years, range 19-77 years) and 90 healthy controls (82 females, 8 male; mean age 36.54 ± 10.89 years, range 22-60 years) were recruited for the current study. Plasma melatonin levels were detected by enzyme-linked immunosorbent assay. RESULTS: Melatonin levels were not significantly different in the plasma of patients with SLE compared with controls (P = 0.026). There was no significant difference regarding plasma melatonin level between SLE patients with nephritis and those without nephritis (P = 0.714); no significant difference was found between less active SLE and more active SLE (P = 0.791). The presence of IgM was associated with melatonin levels (P = 0.031) in SLE patients. CONCLUSIONS: There is no significant difference in plasma melatonin levels between SLE patients and controls. Further studies are needed to elucidate the role of melatonin in SLE.

Electroacupuncture ameliorates spatial learning and memory impairment via attenuating NOX2-related oxidative stress in a rat model of Alzheimer's disease induced by Aß1-42.

Alzheimer's disease (AD) is a chronic neurodegenerative disorder characterized by progressive deterioration of cognition and memory, in which oxidative stress has been played a crucial role in the pathology of AD. Electroacupuncture (EA) is a widely used therapy based on traditional acupuncture combined with modern electrotherapy in Asia. The present study aimed to determine the effects of EA treatment on spatial learning and memory impairment, and to elucidate the status of NOX2-related oxidative stress in a rat model of Alzheimer's disease induced by Beta-amyloid1-42 (Aß1-42). Fifty-six adult female Sprague-Dawley (SD) rats were randomly divided into four groups: sham, sham+EA, AD and AD+EA. The rats in Sham+EA and AD+EA groups were respectively administrated EA treatment at Baihui and yongquan acupoints, once a day for 30 min, lasting for 28 days. The spatial learning and memory functions were assessed by Morris water maze (MWM) test. The activities of total antioxidant capacity (T-AOC), reactive oxygen species (ROS), malondialdehyde (MDA) and 8-hydroxy-2-deoxyguanosine (8-OH-dG) were evaluated. Moreover, the neuronal injury was detected by Nissl staining. Meanwhile, the NeuN expression was examined in the hippocampus, the expression levels of Nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase2(NOX2) was detected by immunofluorescence staining and western blot. The results showed that EA treatment significantly improved spatial learning and memory impairment in rats induced by Aß1-42. Concomitantly, EA treatment markedly restored T-AOC and attenuated the abnormal increase in levels of ROS, MDA and 8-OH-dG in the hippocampus of the AD rats. More notably, EA treatment also effectively ameliorated neuronal injury and counteracted the aberrant increase of NOX2 levels in the hippocampus of AD rats. Our findings suggested that EA is a potential strategy for the treatment of AD, and the possible mechanism is associated with the alleviation of neuronal injury and inhibition of NOX2-related oxidative stress.

Preoperative NLR and PLR in the middle or lower ESCC patients with radical operation.

Neutrophil lymphocyte ratio (NLR) and platelet lymphocyte ratio (PLR) had been analysed in many kind of tumours, but its role of predict the oesophageal squamous cell carcinoma (ESCC) patients' prognosis was not reach a consensus. Relationship between NLR, PLR and ESCC located in the middle or lower segment was evaluated. 317 patients with ESCC who underwent attempted curative oesophagectomy were analysed in this study. 157 and 98 patients had elevated NLR and PLR respectively (NLR >3.3 and PLR >150). The median overall survival time (OS) and disease-free survival (DFS) was 34.1 and 19.2 months respectively. Multivariate analysis found PLR >150 (P = 0.018, HR 1.426, 95%CI 1.063-1.912) accompanied by male, lymphatic metastases, tumour size more than 3 cm, tumour located at middle segment and poor differentiation were associated with significantly worse DFS. Meanwhile, gender, lymphatic metastases, tumour location and differentiation along with PLR >150 (P = 0.003, HR 1.595, 95% CI 1.172-2.170) and NLR>3.3 (P = 0.039, HR 1.367, 95% CI 1.015-1.840) were all independent prognostic factors for OS. Preoperative NLR and PLR might be used as predictive factors in patients with ESCC. For DFS, elevated PLR compared to NLR may have an advantage to indicate poor prognosis.

Unsupervised learning technique identifies bronchiectasis phenotypes with distinct clinical characteristics.

BACKGROUND: Unsupervised learning technique allows researchers to identify different phenotypes of diseases with complex manifestations. OBJECTIVES: To identify bronchiectasis phenotypes and characterise their clinical manifestations and prognosis. METHODS: We conducted hierarchical cluster analysis to identify clusters that best distinguished clinical characteristics of bronchiectasis. Demographics, lung function, sputum bacteriology, aetiology, radiology, disease severity, quality-of-life, cough scale and capsaicin sensitivity, exercise tolerance, health care use and frequency of exacerbations were compared. RESULTS: Data from 148 adults with stable bronchiectasis were analysed. Four clusters were identified. Cluster 1 (n = 69) consisted of the youngest patients with predominantly mild and idiopathic bronchiectasis with minor health care resource use. Patients in cluster 2 (n = 22), in which post-infectious bronchiectasis predominated, had the longest duration of symptoms, greater disease severity, poorer lung function, airway Pseudomonas aeruginosa colonisation and frequent health care resource use. Cluster 3 (n = 16) consisted of elderly patients with shorter duration of symptoms and mostly idiopathic bronchiectasis, and predominantly severe bronchiectasis. Cluster 4 (n = 41) constituted the most elderly patients with moderate disease severity. Clusters 2 and 3 tended to have a greater risk of bronchiectasis exacerbations (P = 0.06) than clusters 1 and 4. CONCLUSION: Identification of distinct phenotypes will lead to greater insight into the characteristics and prognosis of bronchiectasis.

Elevated plasma interleukin-37 levels in systemic lupus erythematosus patients.

This study aims to evaluate the plasma interleukin (IL)-37 levels in systemic lupus erythematosus (SLE) patients, as well as its association with major clinical and laboratory features. Ninety consecutively selected SLE patients and 78 community-based healthy controls were recruited. Plasma IL-37 levels were detected by enzyme-linked immunosorbent assay (ELISA). The major clinical and laboratory data of SLE patients were also recorded. The results showed that IL-37 level was significantly higher in the plasma of patients with SLE compared with controls (p = 0.028). The correlation of plasma IL-37 levels with major clinical and laboratory data of SLE patients was also analyzed, and the results showed that anti-Sm and anti-RNP were negatively associated with plasma IL-37 levels of SLE patients, while C3 was positively associated with plasma IL-37 levels of SLE patients. No significant associations of IL-37 with other clinical and laboratory parameters were observed (all p > 0.05). In conclusion, elevated plasma IL-37 level and its associations with anti-Sm, anti-RNP and C3 in SLE patients suggest that IL-37 may be implicated in this disease.

Pharmacokinetics studies of enrofloxacin injectable in situ forming gel in dogs.

The objective of this study was to evaluate the pharmacokinetic characteristics of enrofloxacin (ENR) injectable in situ gel we developed in dogs following a single intramuscular (i.m.) administration. Twelve healthy dogs were randomly divided into two groups (six dogs per group), then administrated a single 20 mg/kg body weight (b.w.) ENR injectable in situ gel and a single 5 mg/kg b.w. ENR conventional injection, respectively. High-performance liquid chromatography (HPLC) was used to determine ENR plasma concentrations. The pharmacokinetic parameters of ENR injectable in situ gel and conventional injection in dogs are as follows: MRT (mean residence time) (45.59 ± 14.05) h verse (11.40 ± 1.64) h, AUC (area under the blood concentration vs. time curve) (28.66 ± 15.41) µg·h/mL verse (11.06 ± 3.90) µg·h/mL, cmax (maximal concentration) (1.59 ± 0.35) µg/mL verse (1.46 ± 0.07) µg/mL, tmax (time needed to reach cmax ) (1.25 ± 1.37) h verse (1.40 ± 0.55) h, t1/2λz (terminal elimination half-life) (40.27 ± 17.79) h verse (10.32 ± 0.97) h. The results demonstrated that the in situ forming gel system could increase dosing interval of ENR and thus reduced dosing frequency during long-term treatment. Therefore, the ENR injectable in situ gel seems to be worth popularizing in veterinary clinical application.

Sputum bacteriology in steady-state bronchiectasis in Guangzhou, China.

BACKGROUND: The impact of potentially pathogenic micro-organisms (PPMs) on Chinese patients with steady-state bronchiectasis is unknown. METHODS: Peripheral blood and sputum were sampled to determine inflammatory markers and sputum bacterial density. Spirometry and diffusing capacity were measured. Quality of life was assessed using the St George's Respiratory Questionnaire. RESULTS: Of 144 patients with steady-state bronchiectasis, Pseudomonas aeruginosa was isolated in 44 cases (30.6%). Compared with other PPMs, P. aeruginosa had a more pronounced influence on airway inflammation and spirometry, but not on systemic inflammation or quality of life. The impact of PPMs other than P. aeruginosa on clinical indices was similar. Bacterial density was not correlated with most clinical parameters. Factors associated with PPM isolation included bronchiectasis symptoms for ⩾ 10 years (OR 2.13) and ⩾ 4 bronchiectatic lobes (OR 2.82). Having ⩾ 4 exacerbations within 2 years (OR 2.18) and cystic bronchiectasis (OR 2.23) was associated with the colonisation of PPMs, i.e., isolating an identical PPM on at least two occasions within 1 year. CONCLUSION: In patients with steady-state bronchiectasis in Guangzhou, P. aeruginosa is the most common organism causing heightened airway inflammation and poor lung function. PPM isolation or colonisation should be suspected in case of longer duration of symptoms, multilobar bronchiectasis, frequent exacerbation and cystic bronchiectasis.

Study of pharmacokinetics of an in situ forming gel system for controlled delivery of florfenicol in pigs.

To reduce florfenicol (FFC) administration frequency in veterinary use, the drug was currently developed into in situ forming gel. Twelve pigs were randomly divided into two groups (six pigs per group). A single i.m. dose of 40 mg/kg body weight (b.w.) was given to pigs, group one was given FFC in situ forming gel, and group two was given FFC conventional injection. High-performance liquid chromatography (HPLC) was used to determine FFC plasma concentrations. There were significant differences (P < 0.01) between FFC in situ forming gel and conventional injection, in pharmacokinetic parameters MRT (mean retention time) (57.79 ± 2.88) h versus (15.94 ± 1.29) h, AUC (area under the concentration-time curve) (421.54 ± 8.97) µg·h/mL versus (168.16 ± 4.59) µg·h/mL, tmax (time of occurrence of cmax ) (9.00 ± 2.68) h versus (4.33 ± 0.82) h, cmax (maximum plasma concentration) (6.87 ± 0.66) µg/mL versus (12.01 ± 0.66) µg/mL, t1/2λz (terminal elimination half-life) (38.04 ± 2.20) h versus (9.15 ± 2.71) h. The results demonstrated that the in situ forming gel system could shorten dosing interval of FFC and thus achieved less frequent administration during long-term treatment.

Transcriptome profiling of the testis reveals genes involved in spermatogenesis and marker discovery in the oriental fruit fly, Bactrocera dorsalis.

The testis is a highly specialized tissue that plays a vital role in ensuring fertility by producing spermatozoa, which are transferred to the female during mating. Spermatogenesis is a complex process, resulting in the production of mature sperm, and involves significant structural and biochemical changes in the seminiferous epithelium of the adult testis. The identification of genes involved in spermatogenesis of Bactrocera dorsalis (Hendel) is critical for a better understanding of its reproductive development. In this study, we constructed a cDNA library of testes from male B. dorsalis adults at different ages, and performed de novo transcriptome sequencing to produce a comprehensive transcript data set, using Illumina sequencing technology. The analysis yielded 52 016 732 clean reads, including a total of 4.65 Gb of nucleotides. These reads were assembled into 47 677 contigs (average 443 bp) and then clustered into 30 516 unigenes (average 756 bp). Based on BLAST hits with known proteins in different databases, 20 921 unigenes were annotated with a cut-off E-value of 10(-5). The transcriptome sequences were further annotated using the Clusters of Orthologous Groups, Gene Orthology and the Kyoto Encyclopedia of Genes and Genomes databases. Functional genes involved in spermatogenesis were analysed, including cell cycle proteins, metalloproteins, actin, and ubiquitin and antihyperthermia proteins. Several testis-specific genes were also identified. The transcripts database will help us to understand the molecular mechanisms underlying spermatogenesis in B. dorsalis. Furthermore, 2913 simple sequence repeats and 151 431 single nucleotide polymorphisms were identified, which will be useful for investigating the genetic diversity of B. dorsalis in the future.

Validation of the Mandarin Chinese version of the Leicester Cough Questionnaire in bronchiectasis.

BACKGROUND: The Leicester Cough Questionnaire (LCQ) has been validated for assessing cough-specific health status in bronchiectasis. We translated the LCQ into Mandarin Chinese and investigated its validity, reliability and responsiveness. METHODS: The LCQ was translated into Mandarin Chinese using the forward-backward translation procedure. A total of 144 out-patients completed the Mandarin Chinese version of the LCQ (LCQ-MC), the Hospital Anxiety and Depression Scale (HADS) and the St George's Respiratory Questionnaire. Reassessments were performed during exacerbations and at 6 months. Concurrent validation, internal consistency, repeatability and responsiveness were determined. RESULTS: Minor cultural adaptations were made to the wording of LCQ-MC. No other difficulties were found during the translation process, with all items easily adapted to acceptable Mandarin Chinese. The questionnaire was not changed in terms of content layout and the order of the questions. In cognitive debriefing interviews, participants reported that the questionnaire was acceptable, relevant, comprehensive and easy to complete. The LCQ-MC showed good concurrent validity, internal consistency and test-retest reliability. Responsiveness was shown by significant changes in LCQ-MC scores between steady state, the first exacerbation and following 2-week antibiotic treatment (both interval changes, P < 0.01) CONCLUSION: The LCQ-MC is a valid, reliable and responsive instrument for determining cough-specific health status in Chinese bronchiectasis patients.

Novel biotransformation process of podophyllotoxin to 4 ß-sulfur-substituted podophyllum derivates with anti-tumor activity by Penicillium purpurogenum Y.J. Tang.

According to the structure-function relationship of podophyllotoxin (PTOX) and its analogue of 4'- demethylepipodophyllotoxin (DMEP), the 4 ß-substitution of sulfur-containing heterocyclic compounds with a carbon-sulfur bond at 4 position of PTOX or DMEP is an essential modification direction for improving the anti-tumor activity. So, four novel 4 ß-sulfursubstituted podophyllum derivatives (i.e., 4ß -(1,2,4-triazole-3-yl)sulfanyl-4-deoxy-podophyllotoxin (4-MT-PTOX), 4ß-(1,3,4- thiadiazole-2-yl)sulfanyl-4-deoxy-podophyllotoxin (4-MTD-PTOX), 4ß-(1,2,4-triazole-3-yl)sulfanyl-4-deoxy-4' -demethylepipodophyllotoxin (4-MT-DMEP), and 4ß-(1,3,4-thiadiazole-2-yl)sulfanyl-4-deoxy-4'-demethylepipodophyllotoxin (4-MTD-DMEP)) were designed and then successfully biosynthesized in this work. In the novel sulfur-substituted biotransformation processes, PTOX and DMEP was linked with sulfur-containing compounds (i.e., 3-mercapto-1,2,4-triazole (MT) and 2-mercapto-1,3,4-thiadiazole (MTD)) at 4 position of cycloparaffin to produce 4-MT-PTOX (1), 4-MTD-PTOX (2), 4-MT-DMEP (3), and 4-MTD-DMEP (4) by Penicillium purpurogenum Y.J. Tang, respectively, which was screened out from Diphylleia sinensis Li (Hubei, China). All the novel compounds exhibited promising in vitro bioactivity, especially 4-MT-PTOX (1). Compared with etoposide (i.e., a 50 % effective concentration [EC(50)] of 25.72, 167.97, and 1.15 M), the EC(50) values of 4-MT-PTOX (1) against tumor cell line BGC-823, A549 and HepG2 (i.e., 0.28, 0.76, and 0.42 M) were significantly improved by 91, 221 and 2.73 times, respectively. Moreover, the EC(50) value of 4-MT-PTOX (1) against the normal human cell line HK-2 (i.e., 182.4 µM) was 19 times higher than that of etoposide (i.e., 9.17 µM). Based on the rational design, four novel 4 ß-sulfur-substituted podophyllum derivatives with superior in vitro anti-tumor activity were obtained for the first time. The correctness of structure-function relationship and rational drug design was strictly demonstrated by the in vitro biological activity tests.

Enhanced differentiation of human adipose tissue-derived stromal cells into insulin-producing cells with glucagon-like peptide-1.

Type 1 diabetes mellitus (T1DM) is mainly caused by reduction of the endogenous insulin secretion due to autoimmune destruction of pancreatic ß cells, and a promising therapeutic approach for T1DM is pancreas and islet cell replacement. The major obstacle is the limited source of insulin-producing cells. Here, we report an efficient approach to induce human adipose-derived stromal cells (hADSCs) to differentiate into insulin-producing cells, with glucagon-like peptide-1 (GLP-1). hADSCs were successfully isolated from the adipose tissue, with adipogenic and osteogenic differentiation potency. Islet-like cell clusters formed in the culture, which was enhanced with the treatment of GLP-1. Reverse transcription polymerase chain reaction analysis showed the expression of the pancreas-related genes in the differentiated cells, such as pdx-1, ngn3, insulin, glucagon, somatostatin, glucokinase n and glut2. Immunocytochemical analysis showed that the induced cells co-expressed insulin, C-peptide and PDX-1. The GLP-1 receptor was present in the differentiated cells. In addition, flow cytometry analysis and ELISA showed that, in the presence of GLP-1, the percentage of insulin-producing cells was increased from 5.9% to 28.0% and the release of insulin increased from 9.53±0.7 pmol/106 cells to 15.86±1.3 pmol/106 cells. Insulin was released in response to glucose stimulation in a manner comparable to that of adult human islets. These results indicated that hADSCs isolated from adipose tissues can be induced to differentiate into insulin-producing cells, which is further enhanced with the treatment of GLP-1. These findings confirm that the differentiation of hADSCs to insulin-producing cells is indeed possible and indicate that the differentiated insulin-producing cells can be used as a potential source for transplantation into patients with T1DM.

The effect of the expression of vascular endothelial growth factor (VEGF)-C and VEGF receptor-3 on the clinical outcome in patients with gastric carcinoma.

AIMS: We aimed to investigate the relationship among VEGF-C/VEGFR-3 expression, lymphatic metastasis and patient prognosis in gastric carcinoma. MATERIAL AND METHODS: VEGF-C and VEGFR-3 expression in gastric carcinoma tissues obtained from 204 patients who underwent curative gastrectomy (105 cases presented with lymph node metastasis and 99 cases without metastasis) was examined immunohistochemically. There was no significant difference in the other clinicopathologic variables except for postoperative pathological tumor stage (pT) and TNM stage between the two groups. The results were statistically processed. RESULTS: The results showed that VEGF-C was located mainly in the cytoplasm of tumor cells and VEGFR-3 was found predominantly in the endothelium of lymphatic vessels. VEGF-C and VEGFR-3 expression was more frequent in gastric carcinoma tissues than that in normal gastric tissues, 54.90% and 35.29% respectively, which revealed that the expression of VEGF-C and VEGFR-3 was significantly stronger in patients with lymph node metastasis than in those without metastasis. Patients who had positive staining for VEGF-C showed significantly less favorable survival rates compared with patients who had negative staining for VEGF-C. The survival rates of patients who had positive staining for VEGFR-3 also were significantly lower compared with patients who had negative staining for VEGFR-3. Patients who had positive staining for both VEGF-C and VEGFR-3 exhibited the most unfavorable prognosis. Multivariate analysis demonstrated that the expression of VEGF-C and VEGFR-3 was an independent prognostic determinant. In addition, faint to moderate VEGF-C expression was detected in normal gastric epithelial cells (18/204, 8.9%). CONCLUSIONS: VEGF-C and VEGFR-3 expression could serve as a prognostic biomarker in patients with gastric carcinoma.

Minichromosome maintenance proteins 2, 3 and 7 in medulloblastoma: overexpression and involvement in regulation of cell migration and invasion.

Minichromosome maintenance (MCM) proteins 2-7 are important in DNA replication licensing. Functional roles beyond licensing are speculated. In addition, significances in medulloblastoma (MB) remain unclear. In this study, we showed the frequent deregulation of MCM2 and MCM3 expression in 7 MB cell lines and 31 clinical samples. Moreover, DAOY and ONS76 and the clinical samples expressed elevated MCM7 transcripts with genomic gain of the gene. Immunopositivity restricted to tumor cells was found in 41, 37 and 53 out of 73 MB cases for MCM2, MCM3 and MCM7, respectively. High-MCM3 expression was associated with poor prognosis. Knockdowns of these MCMs significantly inhibited anchorage-dependent and -independent MB cell growth. The inhibition of MCM3 expression by small interfering RNA knockdown was related to G1 arrest with reduced cyclin A expression, whereas the MCM2- and MCM7-knocked-down cells arrested at G2/M with increased cyclin A expression. Interestingly, we demonstrated the links of these MCMs with cell migration and invasion using wound-healing and Transwell migration/invasion assays. Exogenous overexpression of MCM2, MCM3 and MCM7 increased anchorage-independent cell growth, and also cell migration and invasion capabilities in MB cells. The knockdown reduced the number of filopodial cells and the cells with intense stress fibers by blocking cdc42 and Rho activation. Taken together, deregulation of MCM2, MCM3 and MCM7 expression might be involved in MB tumorigenesis and we revealed undefined roles of these MCMs in control of MB cell migration and invasion.

Decreased detoxification genes and genome size make the human body louse an efficient model to study xenobiotic metabolism.

The human body louse, Pediculus humanus humanus, has one of the smallest insect genomes, containing ∼10 775 annotated genes. Annotation of detoxification [cytochrome P450 monooxygenase (P450), glutathione-S-transferase (GST), esterase (Est) and ATP-binding cassette transporter (ABC transporter)] genes revealed that they are dramatically reduced in P. h. humanus compared to other insects except for Apis mellifera. There are 37 P450, 13 GST and 17 Est genes present in P. h. humanus, approximately half the number found in Drosophila melanogaster and Anopheles gambiae. The number of putatively functional ABC transporter genes in P. h. humanus and Ap. mellifera are the same (36) but both have fewer than An. gambiae (44) or Dr. melanogaster (65). The reduction of detoxification genes in P. h. humanus may be a result of this louse's simple life history, in which it does not encounter a wide variety of xenobiotics. Neuronal component genes are highly conserved across different insect species as expected because of their critical function. Although reduced in number, P. h. humanus still retains at least a minimum repertoire of genes known to confer metabolic or toxicokinetic resistance to xenobiotics (eg Cyp3 clade P450s, Delta GSTs, B clade Ests and B/C subfamily ABC transporters), suggestive of its high potential for resistance development.