A novel mutation in GPR68 causes hypomaturation amelogenesis imperfecta.

OBJECTIVES: To identify the genetic cause of a Chinese family with hypomaturation amelogenesis imperfecta (AI) and to characterize the structure of GPR68 mutated enamel in order to develop a deeper understanding of the role of the GPR68 protein during the intricate process of amelogenesis. DESIGN: One Chinese family with generalized hypomaturation AI was recruited. Two of the third molars from the proband were subjected to scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDX). Whole exome sequencing (WES) was performed, and the identified mutation was confirmed by Sanger sequencing. Bioinformatics studies were further conducted to analyze the potential deleterious effects of the mutation. RESULTS: The proband presented with a hypomaturation AI phenotype, characterized by fragile and discolored enamel surface. The AI enamel showed prismatic structure, which was sporadically obscured by areas of amorphous material and porous structure. EDX analysis showed the proband's enamel demonstrated a significant decrease in calcium and phosphorus content and a significant increase in oxygen compared with normal enamel. A novel homozygous mutation of G protein-coupled receptor 68 (GPR68) (c .149 T > A, p.Ile50Asn) was identified in the proband. Bioinformatics analysis indicated that the mutation site displayed a high level of evolutionary conservation among species, and the mutation might impact the stability and conformation of the protein. CONCLUSION: The novel homozygous GPR68 mutation resulted in hypomaturation AI. We first described the effect of GPR68 mutation on enamel structure. Our results provide new genetic evidence that mutations involved in GPR68 contribute to hypomaturation AI.

Mitochondrial dysfunction of peripheral blood mononuclear cells is associated with lung carcinogenesis.

The composition, quantity, and function of peripheral blood mononuclear cells (PBMCs) are closely correlated with tumorigenesis. However, the mechanisms of PBMCs in lung cancer are not clear. Mitochondria are energy factories of cells, and almost all cellular functions rely on their energy metabolism level. The present study aimed to test whether the mitochondrial function of PBMCs directly determines their tumor immune monitoring function. We recruited 211 subjects, including 105 healthy controls and 106 patients with recently diagnosed with lung cancer. The model of lung carcinogenesis induced by BaP was used in animal experiment, and the Bap carcinogenic metabolite, Benzo(a)pyren-7,8-dihydrodiol-9,10-epoxide (BPDE), was used in cell experiment. We found that mitochondrial function of PBMCs decreased significantly in patients with new lung cancer, regardless of age. In vivo, BaP caused PBMC mitochondrial dysfunction in mice before the appearance of visible malignant tissue. Moreover, mitochondrial function decreased significantly in mice with lung cancers induced by BaP compared to those without lung cancer after BaP intervention. In vitro, BPDE also induced mitochondrial dysfunction and reduced the aggressiveness of PBMCs toward cancer cells. Furthermore, the changes in mitochondrial energy metabolism gene expression caused by BPDE are involved in this process. Thus, the mitochondrial function of PBMCs is a potential prognostic biomarker or therapeutic target to improve clinical outcomes in patients with lung cancer.

Effect of lemon essential oil on halitosis.

OBJECTIVES: To evaluate the effect of lemon essential oil (LEO) on salivary bacteria and volatile sulfur compound (VSC) production of patients with halitosis. MATERIALS AND METHODS: Saliva of five patients with halitosis was collected, after adding different concentrations (0.563-9 mg/ml) of LEO, detecting the growth of salivary bacteria, the formation of biofilm, and VSC production, and compare the difference of different concentrations of LEO on bacterial growth and VSC production. 48 volunteers were randomly divided into 4 groups. After gargling with LEO, cetylpyridinium chloride (CPC), chlorhexidine (CHX), and hydrogen peroxide (H2 O2 ) separately measure changes of VSC production and pH values at 30, 45, 60, 90, and 120 min and then compare the differences at different time points within group. RESULTS: Compared with the negative control group, under subinhibitory concentrations of LEO (0.563-2.25 mg/ml), the biofilm formation and VSC production of salivary bacteria in LEO group were significantly inhibited (p < 0.05). Compared with the baseline, the VSC production of subjects decreased after rinsing with the LEO in 60 min (p < 0.05). After gargling with LEO, the pH value rose significantly in 30 min and reverted to the baseline level at 120 min (p < 0.05). CONCLUSIONS: Lemon essential oil can inhibit the growth of salivary bacteria and reduce VSC production of patients with halitosis.

Identification of potential prognostic biomarkers among gene models for coiled-coil domain-containing family members in hepatocellular carcinoma elucidates their influence on the hypoxia pathway and immune microenvironment.

Background: The family of coiled-coil domain-containing (CCDC) proteins participates in a wide range of physiological functions and plays a pivotal role in governing the invasion and metastasis of malignant tumor cells. Nonetheless, the precise mechanism governing the interaction among the immune microenvironment, hypoxia pathway, and proliferation in hepatocellular carcinoma (HCC) remains elusive. In this study, our objective was to identify the prognostic significance of CCDC family genes in HCC. Methods: We conducted an analysis of RNA-seq data from HCC patients sourced from The Cancer Genome Atlas (TCGA) database. Our analysis involved comparing the expression profiles of 168 CCDC family genes between tumor and normal tissues to identify differentially expressed genes (DEGs). The prognostic value of these genes was verified using overall survival (OS) data from TCGA-LIHC patients, employing Univariate and multivariate Cox proportional hazards regression models and Kaplan-Meier plots. Subsequently, we constructed a prognostic signature known as the CCDC score and validated it using additional datasets (ICGC-LIRI-JP and GSE14520). Additionally, we performed functional enrichment analysis and conducted an assessment of the tumor immune microenvironment (TIME). Results: We identified 34 DEGs of the CCDC family. Among them, six DEGs (CCDC6/22/51/59/132/134) were upregulated and associated with poor prognosis. Higher CCDC score was an independent predictor of poor OS in TCGA-HCC patients (P<0.001, HR =2.37), which was validated in the ICGC-LIRI-JP (P=0.021, HR =2.15) and GSE14520 (P=0.002, HR =2.23) datasets. Functional enrichment analysis showed that hypoxia pathway genes were enriched in the high CCDC score group. Furthermore, immune microenvironment analysis demonstrated that high CCDC score was associated with a suppressed TIME caused by the extrinsic immune escape. Conclusions: The CCDC score, derived from six CCDC genes, exhibits remarkable expression levels in liver cancer and holds promise as an independent prognostic indicator. Our bioinformatics analysis revealed a high CCDC score is strongly associated with activation of the hypoxia pathway and an immunosuppressive tumor microenvironment in HCC. This profound finding may serve as a cornerstone for innovative targeted drug therapies and pave the way for further investigations into the underlying mechanisms of CCDC-related carcinogenesis in liver cancer.

Effect of Lemon Essential Oil Microemulsion on the Cariogenic Virulence Factor of Streptococcus mutans via the Glycolytic Pathway.

PURPOSE: To investigate the effects and mechanisms of lemon essential oil products on dental caries prevention. MATERIALS AND METHODS: Lemon essential oil microemulsions (LEOM) with concentrations of 1/8 minimum inhibitory concentration (MIC), 1/4 MIC, and 1/2 MIC were applied to S. mutans at concentrations of 0.2%, 1%, and 5% glucose, respectively. Changes in acid production capacity of S. mutans were measured based on changes in pH. The effect of the reductive coenzyme I oxidation method on LDH activity was examined. The effect of lemon essential oil microemulsion on the expression of the lactate dehydrogenase gene (ldh) was detected by a quantitative real-time polymerase chain reaction. RESULTS: Lemon essential oil microemulsion at 1/2 MIC concentration reduced the environmental pH value at different glucose concentrations, compared to those observed in the control group (p < 0.05). LDH activity of S. mutans was decreased at three subinhibitory concentrations of lemon essential oil microemulsions (p < 0.05). The effect of lemon essential oil microemulsions on S. mutans LDH activity and bacterial acid production were positively correlated (r = 0.825, p < 0.05). Lemon essential oil microemulsion at 1/2 MIC concentration downregulated the expression of the ldh gene of S. mutans at different glucose concentrations (p < 0.05). In different glucose environments, lemon essential oil microemulsions at subminimum inhibitory concentrations can inhibit the acid production of S. mutans by reducing ldh expression and LDH activity in the glycolytic pathway, proving its anti-caries potential. CONCLUSIONS: LEOM can effectively prevent dental caries and maintain the microecological balance of the oral environment.

Resveratrol Ameliorates High Altitude Hypoxia-Induced Osteoporosis by Suppressing the ROS/HIF Signaling Pathway.

Hypoxia at high-altitude leads to osteoporosis. Resveratrol (RES), as an antioxidant, has been reported to promote osteoblastogenesis and suppress osteoclastogenesis. However, the therapeutic effect of RES against osteoporosis induced by high-altitude hypoxia remains unclear. Thus, this study was intended to investigate the potential effects of RES on high-altitude hypoxia-induced osteoporosis both in vivo and in vitro. Male Wistar rats were given RES (400 mg/kg) once daily for nine weeks under hypoxia, while the control was allowed to grow under normoxia. Bone mineral density (BMD), the levels of bone metabolism-related markers, and the changes on a histological level were measured. Bone marrow-derived mesenchymal stem cells (BMSCs) and RAW264.7 were incubated with RES under hypoxia, with a control growing under normoxia, followed by the evaluation of proliferation and differentiation. The results showed that RES inhibited high-altitude hypoxia-induced reduction in BMD, enhanced alkaline phosphatase (ALP), osteocalcin (OCN), calcitonin (CT) and runt-related transcription factor 2 (RUNX2) levels, whereas it reduced cross-linked carboxy-terminal telopeptide of type I collagen (CTX-I) levels and tartrate-resistant acid phosphatase (TRAP) activity in vivo. In addition, RES attenuated histological deteriorations in the femurs. In vitro, RES promoted osteoblastogenesis and mineralization in hypoxia-exposed BMSCs, along with promotion in RUNX2, ALP, OCN and osteopontin (OPN) levels, and inhibited the proliferation and osteoclastogenesis of RAW264.7. The promotion effects of RES on osteoblastogenesis were accompanied by the down-regulation of reactive oxygen species (ROS) and hypoxia inducible factor-1α (HIF-1α) induced by hypoxia. These results demonstrate that RES can alleviate high-altitude hypoxia-induced osteoporosis via promoting osteoblastogenesis by suppressing the ROS/HIF-1α signaling pathway. Thus, we suggest that RES might be a potential treatment with minimal side effects to protect against high-altitude hypoxia-induced osteoporosis.

Bee venom protects against pancreatic cancer via inducing cell cycle arrest and apoptosis with suppression of cell migration.

Background: Pancreatic cancer seriously threatens human health. Bee venom is a mixture of enzymes, peptides, and amines. Due to its biological activity, bee venom is widely used as an anti-inflammatory agent and pain reliever. However, little is known about the effect of bee venom on pancreatic cancer. Methods: Firstly, the Cell Counting Kit-8 (CCK-8) assay was conducted to analyze the cytotoxicity of bee venom on PANC-1 and AsPC-1 cells. Then, we evaluated the cell cycle and apoptosis by flow cytometry and the terminal deoxynucleotidyl transferase (TdT) dUTP Nick-End Labeling (TUNEL) assay. In addition, cell migration was analyzed by the cell scratch test and Transwell assay. Western blot was performed to assess the expression of proteins involved in the regulation of cell cycle arrest and apoptosis. Results: Results demonstrated that bee venom significantly suppressed cell proliferation via inducing cell cycle arrest and apoptosis with suppression of cell migration. Bee venom induced S phase arrest and ameliorated the protein expression of cyclins and cyclin-dependent kinases (CDKs). At the same time, bee venom can activate the p53-p21 pathway. Experimental data also showed that bee venom induced cell apoptosis and impeded cell migration. Conclusions: The present study revealed that bee venom could effectively inhibit tumor progression in pancreatic cancer cells, indicating the possibility of bee venom as an anti-tumor drug in pancreatic cancer.

Role of mitochondrial reactive oxygen species in homeostasis regulation.

Mitochondria are the main source of reactive oxygen species (ROS) in cells. Early studies have shown that mitochondrial reactive oxygen species (mROS) are related to the occurrence and adverse outcomes of many diseases, and are thus regarded as an important risk factor that threaten human health. Recently, increasing evidence has shown that mROS are very important for an organism's homeostasis. mROS can regulate a variety of signaling pathways and activate the adaptation and protection behaviors of an organism under stress. In addition, mROS also regulate important physiological processes, such as cell proliferation, differentiation, aging, and apoptosis. Herein, we review the mechanisms of production, transformation, and clearance of mROS and their biological roles in different physiological processes.

[Resveratrol attenuates cardiac function impairment in plateau hypobaric hypoxia rats].

Objective: To study the protective effects of resveratrol (RSV) on cardiac function in rats with high altitude hypobaric hypoxia and its mechanisms. Methods: Thirty-six rats were randomly divided into control group, hypobaric hypoxia group (HH) and hypobaric hypoxia + RSV group (HH+RSV) according to the random number, 12 rats in each group. Rats in the HH and HH+RSV groups were subjected to chronic long-term high altitude hypobaric hypoxia intervention for 8 weeks in a hypobaric chamber at a simulated altitude of 6 000 m for 20 h / d. The rats of HH + RSV were fed with RSV at a dose of 400 mg/(kg·d). The rats were tested once a week for body weight and twice a week for food intake. Before execution, the rats were tested by blood cell analyzer for routine blood parameters and echocardiogram for cardiac function parameters in each group. The routine blood indexes of each group were measured by blood cell analyzer, the cardiac function indexes of each group were measured by echocardiography, myocardial hypertrophy was evaluated by HE staining, myocardial tissue reactive oxygen levels were evaluated by dihydroethidium (DHE) staining. Oxidative stress was evaluated by serum and myocardial tissue total antioxidant capacity (T-AOC), superoxide dismutase activity (SOD) and malondialdehyde (MDA) content. Results: Compared with the C group, the body mass and food intake of rats were decreased significantly (P＜0.05) in HH group, while compared with the C group, RSV had no significant effects on the body mass and food intake of rats in the HH+RSV group (P＞0.05). Compared with the C group, the levels of erythrocytes and hemoglobin of rats in the HH group were increased significantly (P＜0.05), while the platelet concentration was decreased significantly(P＜0.05); compared with the HH group, the erythrocyte and hemoglobin levels were decreased significantly (P＜0.05) and platelet concentration was increased significantly(P＜0.05) in rats of the HH+RSV group. Compared with the C group, the cardiac coefficient, myocardial fiber diameter and thickness were significantly increased in the HH group (P＜0.05); compared with the HH group, the cardiac coefficient and myocardial fiber thickness were significantly decreased in the HH+RSV group (P＜0.05). Echocardiographic analysis showed a significant increase in ventricular wall thickness (P＜0.05) and a significant decrease in ejection fraction and cardiac output (P＜0.05) in the HH group compared with the C group, and a significant decrease in ventricular wall thickness and a significant improvement in cardiac function (P＜0.05) in the HH+RSV group compared with the HH group. The results of DHE staining showed that myocardial tissue reactive oxygen levels were increased significantly in the HH group compared with the C group (P＜0.05); myocardial tissue reactive oxygen levels were significantly restored in the HH+RSV group compared with the HH group (P＜0.05). The oxidative/antioxidant results showed that the serum and myocardial T-AOC and SOD activities were decreased significantly (P＜0.05) and the MDA level was increased significantly (P＜0.05) in the HH group compared with the C group; the serum and myocardial T-AOC and SOD activities were increased significantly (P＜0.05) and the MDA level was decreased significantly(P＜0.05) in the HH+RSV group compared with the HH group. Conclusion: Long-term plateau hypobaric hypoxia exposure leads to myocardial hypertrophy and reduced cardiac function in rats. Resveratrol intervention significantly improves myocardial hypertrophy and cardiac function in rats caused by altitude hypobaric hypoxia exposure, which is closely related to reducing of reactive oxygen species and improving myocardial oxidative stress levels.

A Multi-Sensor Based Roadheader Positioning Model and Arbitrary Tunnel Cross Section Automatic Cutting.

Autonomous posture detection and self-localization of roadheaders is the key to automatic tunneling and roadheader robotization. In this paper, a multi-sensor based positioning method, involving an inertial system for altitude angles measurement, total station for coordinate measurement, and sensors for measuring the real-time length of the hydraulic cylinder is presented for roadheader position measurement and posture detection. Based on this method, a positioning model for roadheader and cutter positioning is developed. Additionally, flexible trajectory planning methods are provided for automatic cutting. Based on the positioning model and the trajectory planning methods, an automatic cutting procedure is proposed and applied in practical tunneling. The experimental results verify the high accuracy and efficiency of both the positioning method and the model. Furthermore, it is indicated that arbitrary shapes can be generated automatically and precisely according to the planned trajectory, employing the automatic cutting procedure. Therefore, unmanned tunneling can be realized by employing the proposed automatic cutting process.

Laparoscopic pancreaticoduodenectomy for metastatic pancreatic melanoma: A case report.

RATIONALE: Pancreatic metastases from other malignant tumors are an uncommon clinical condition and account for approximately 2% of all pancreatic malignancies. The most common primary malignancy that metastasizes to pancreas is renal cell cancer. We reported a rare clinical case of metastatic melanoma to pancreas who underwent a successful laparoscopic pancreaticoduodenectomy (LPD) at our department. PATIENT CONCERNS: A 54-year-old Chinese man complaining an unexplained jaundice was found to have a pancreatic mass and he was diagnosed with cutaneous melanoma (CM) 6 years ago. DIAGNOSES: Contrast-enhanced computed tomography (CECT) revealed a solid hypovascular mass measuring about 3.1 × 2.4 cm localized at the junction of pancreatic head and uncinate process, which compressed the lower common bile duct resulting in expansion of the upstream bile ducts. INTERVENTIONS: We performed an LPD and regional lymphadenectomy on this patient. OUTCOMES: This patient was discharged home on postoperative day 19. Postoperative pathological results revealed a malignant melanoma with negative margins. Immunohistochemical (IHC) findings also suggested a malignant pancreatic tumor accompanied by necrosis and pigmentation, which confirmed the pathological diagnosis. Immunoreactivity was strongly positive for anti-S-100 protein (+++) and positive for anti-Vimentin (+). The cancer cells were negative for CEA, CK8/18, P53, Violin, CK19, SMA with Ki-67 over 40%. So this pancreatic mass was proved to be a metastatic pancreatic melanoma from the primary cutaneous lesion. After LPD, this patient was followed up by readmission to hospital every 2 month in the first half year. The serum bilirubin and tumor markers such as CA199 were normal. CECT and did not find any newly developed neoplasm at the pancreas or metastasis at other organs. At the last follow-up at 6 months after LPD, the patient's general condition was acceptable and the physical examination and imaging studies revealed no significant findings of melanoma. LESSONS: Metastatic pancreatic tumors are often associated with well-defined margins, tumor necrosis, enhancement, and distant metastases without pancreatic duct dilatation and parenchymal atrophy. As the most common type of metastatic pancreatic tumor, renal cell cancers tend to have higher attenuation values than that of primary pancreatic cancer, while they had similar attenuation values on the portal phase. Primary pancreatic cancer was always associated with an elevated CA199, total bilirubin, and fasting plasma glucose levels. Surgical resection for metastases to pancreas should be aggressively considered in selected patients due to its unique value of providing palliation and a chance to cure. For patients with unresectable lesions, new therapeutic protocols should be recommended such as the combination of BRAF with MEK inhibitor and PD-1 blocker with or without ipilimumab.

Two nonsense somatic mutations in MEN1 identified in sporadic insulinomas.

Insulinomas are functional pancreatic neuroendocrine tumors that cause hypoglycemia and severe morbidity. The aim of our study was to identify gene mutations responsible for tumorigenesis of sporadic insulinoma. Whole exome sequencing analysis was performed on tumors and paired peripheral blood from three patients with insulinomas. After initial analysis, somatic mutations were obtained and a deleterious protein product was further predicted by various bioinformatic programs. Whole exome sequencing identified 55 rare somatic mutations among three insulinoma patients, including MEN1 gene nonsense mutations (c. 681C>G; p.Tyr227* in exon 4 of MEN1 and c. 346G>T; p.Glu116* in exon 2 of MEN1) in two different tumor samples. The mutations resulted in a significant truncation of the protein and a non-functional gene product, which was involved in defective binding of menin to proteins implicated in genetic and epigenetic mechanisms. Our results extend the growing list of pathogenic MEN1 mutations in sporadic cases of insulinoma.

Effects of LncRNA-HOST2 on cell proliferation, migration, invasion and apoptosis of human hepatocellular carcinoma cell line SMMC-7721.

The present study explored the effect of long non-coding RNA-human ovarian cancer-specific transcript 2 (LncRNA-HOST2) on cell proliferation, migration, invasion and apoptosis of human hepatocellular carcinoma (HCC) cell line SMMC-7721. HCC tissues and adjacent normal tissues from 162 HCC patients were collected. The HCC cell lines were assigned into the control group (regular culture), negative control (NC) group (transfected with siRNA) and experimental group (transfected with Lnc-HOST2 siRNA). Quantitative real-time PCR (qRT-PCR) was used to detect the expression of LncRNA-HOST2. Cell proliferation was detected by CCK-8 and colony-forming assays, cell apoptosis by flow cytometry and cell migration by Scratch test. Transwell assay was used to evaluate cell migration and invasion abilities. LncRNA-HOST2 expression in the HCC tissues increased 2-10 times than that in the adjacent normal tissues. Compared with the HL-7702 cell line, LncRNA-HOST2 expression in HepG2, SMMC-7721 and Huh7 cell lines was all up-regulated, but the SMMC-7721 cell had the highest Lnc-HOST2 expression. The LncRNA-HOST2 expression in the experimental group was down-regulated as compared with the control and NC groups. In comparison with the control and NC groups, cloned cells reduced, cell apoptosis increased, clone-forming ability weakened and inhibitory rate of colony formation increased in the experimental group. The cells migrating and penetrating into the transwell chamber were fewer in the experimental group than those in the control and NC groups. The experimental group exhibited slow wound healing and decreased cell migration area after 48 h. These findings indicate that LncRNA-HOST2 can promote cell proliferation, migration and invasion and inhibit cell apoptosis in human HCC cell line SMMC-7721.

Downregulation of MicroRNA-145 Caused by Hepatitis B Virus X Protein Promotes Expression of CUL5 and Contributes to Pathogenesis of Hepatitis B Virus-Associated Hepatocellular Carcinoma.

BACKGROUND: Hepatitis B viral infection-induced hepatocellular carcinoma (HCC) is a major threat to human health in China. Hepatitis B virus X protein (HBX), an HBV protein, has been reported to be involved in regulating the cellular activities of the host cells and is responsible for HCC oncogenesis. METHODS AND RESULTS: In this study, we performed real-time PCR in tumor tissue samples collected from 53 HCC patients (25 HBV-positive cases and 28 HBV-negative cases) to screen the candidate miRNAs that have previously been reported to be aberrantly expressed in HBV-associated HCC and found that miR-145 was significantly downregulated. The following computational analysis identified CUL5 and RAB5C as virtual targets of miR-145, whereas only CUL5 was verified as a validated target gene of miR-145 in liver cells via luciferase reporter assay. In line with this result, we found that both the mRNA and protein expression levels of CUL5 were significantly higher in HBV-positive than in HBV-negative HCC. An in vitro experiment demonstrated a significant decrease in the expression of miRNA-145, a substantial increase in the mRNA and protein expression of CUL5, and an enhanced proliferation of HBX over-expressing HepG2 cells compared with the control. In HepG2.2.15, we found significant decreases in both the expression of CUL5 and the cell growth rate of H cells transfected with 60 nM miR-145 mimics compared with the scramble controls. CONCLUSION: HBV infection promotes cell growth, at least partially, through the HBX-induced downregulation of miRNA-145 expression, which is responsible for the oncogenesis of HBV-associated HCC.

Voltage-gated sodium channels were differentially expressed in human normal prostate, benign prostatic hyperplasia and prostate cancer cells.

Voltage-gated sodium channels (VGSCs) are expressed not only in excitable cells but also in numerous metastatic cells, particularly in certain types of cancer cells. In some types of cancer, including prostate cancer, the expression of VGSCs is associated with cancer migration, invasion and metastasis in vivo. However, the detailed expression profiles of VGSC α subunits in normal human prostate, in prostatic hyperplasia and prostatic cancer remain controversial. In the present study, quantitative polymerase chain reaction was used to systematically detect all subtypes of VGSC α subunits in normal human prostate, benign prostatic hyperplasia (BPH) and prostate cancer cells. The expression profile of VGSC α subunits was observed to differ between these cell types. Nav1.5 was the major isoform expressed in normal human prostate tissue, while Nav1.5 and Nav1.2 were the predominant isoforms in BPH tissue. However, in PC-3 and LNCaP cells, two typical prostate cancer cell lines, Nav1.6 and Nav1.7 were abundantly expressed. By comparing the relative expression levels of Nav1.5, Nav1.6 and Nav1.7 in these cells, the mRNA levels of Nav1.6 and Nav1.7 were identified to be 6- to 27-fold higher in PC-3 and LNCaP cells than in either normal or BPH samples (P<0.05); however, Nav1.5 mRNA levels were relatively lower compared with those of Nav1.6 or Nav1.7 in all cells analyzed. To confirm whether Nav1.6 and Nav1.7 expression in cancer cells was functional, a patch-clamp technique was used to record whole-cell currents. A tetrodotoxin-sensitive sodium current was successfully recorded in PC-3 cells, but not in LNCaP cells. It was concluded that although all types of VGSC α subunits exhibited low expression levels in normal prostate and BPH cells, both Nav1.6 and Nav1.7 were significantly upregulated in the prostate cancer cell lines, suggesting these subtypes may be potential diagnostic markers and therapeutic targets for certain types of prostate cancer in humans.

RNA interference-mediated silencing of VEGF and bFGF suppresses endostatin secretion in pancreatic carcinoma cells.

Pro-angiogenic factors [vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF)] and anti-angiogenic factors (endostatin) play important roles in the progression of pancreatic cancer. The purpose of the present study was to investigate the knockdown effect by either VEGF or bFGF siRNA on the expression and secretion of endostatin in pancreatic carcinoma cells. Pancreatic carcinoma cell lines (sw1990, Panc-1 and PCT-3) were treated with VEGF and bFGF siRNA. The expression of VEGF, bFGF and endostatin in pancreatic carcinoma cell lines was determined by reverse transcription-polymerase chain reaction (RT-PCR) and western blot analysis. Secretion of endostatin was measured by enzyme-linked immunosorbent assay (ELISA). bFGF and VEGF siRNA significantly reduced the expression of bFGF and VEGF mRNA, respectively, but did not affect mRNA and protein expression of endostatin in pancreatic carcinoma cell lines. However, secretion of endostatin in PCT-3, Panc-1 and sw1990 cells was significantly inhibited by bFGF and VEGF siRNA. This study demonstrated that pro-angiogenic factors (VEGF and bFGF) differentially modulate expression and secretion of anti-angiogenic factors (endostatin). This result may have important implications in the anti-angiogenesis therapy in pancreatic cancer.

[The significance of VEGF siRNA and bFGF siRNA in invasion and proliferation of pancreatic carcinoma cell].

OBJECTIVE: To investigate the relationship between expression of angiogenic factors and invasion and proliferation of pancreatic cancer cell. METHODS: The three pancreatic cancer cell lines of SW1990, Panc-1 and PCT-3 were divided into four groups respectively: control group, VEGF siRNA group, bFGF siRNA group and VEGF siRNA + bFGF siRNA group. The expression and the secretion of VEGF and bFGF in the three cell lines were inhabited by VEGF siRNA and bFGF siRNA. The proliferation and the invasion of the three cell lines were determined by CCK-8 and Boyden Chamber invasion tests. RESULTS: Expressions of VEGF and bFGF in three cell lines were significantly inhibited by VEGF siRNA and bFGF siRNA. The proliferation was inhabited by VEGF siRNA and bFGF siRNA in SW1990 and Panc-1 (P < 0.05), while was not in PCT-3 (P > 0.05). The invasion was inhabited significantly by VEGF siRNA and bFGF siRNA in the three cell lines (P < 0.05). Combination of VEGF siRNA and bFGF siRNA resulted in more efficient influence in inhibition of invasion in Panc-1 and proliferation in PCT-3 and SW1990 than VEGF siRNA or bFGF siRNA individually. CONCLUSION: The decreased expression of VEGF and bFGF can inhabited the ability of invasion and proliferation of pancreatic cancer cell.

[Expression of angiogenic factors in pancreatic carcinoma cell and their significance].

OBJECTIVE: To investigate the relationship between expression of angiogenic factors and invasion and proliferation in pancreatic carcinoma cell. METHODS: Expressions and secretions of angiogenic factors in pancreatic carcinoma cell lines were determined by RT-PCR and ELISA. Proliferation and invasion of pancreatic carcinoma cell lines were determined by CCK-8 and Boyden Chamber invasion tests. RESULTS: Pancreatic carcinoma cell lines showed significantly different ability of invasion and proliferation. Intracellular VEGF expressions of SW1990, Capan-1, Aspc-1, MiaPaCa-2, Panc-1 and PCT-3 were (152.9 +/- 0.6), (224.1 +/- 60.3), (239.2 +/- 2.1), (19.3 +/- 0.7), (165.6 +/- 34.3), and (18.1 +/- 1.4) pg/(10(6)cell.24 h). Extracellular VEGF expressions of SW1990, Capan-1, Aspc-1, MiaPaCa-2, Panc-1 and PCT-3 were (1331.1 +/- 67.8), (3902.6 +/- 79.7), (2657.3 +/- 51.9), (1498.3 +/- 4.8), (4696.8 +/- 45.5), and (1200.5 +/- 42.2) pg/(10(6)cell.24 h). Intracellular bFGF expressions of SW1990, Capan-1, Aspc-1, MiaPaCa-2, Panc-1 and PCT-3 were (66.1 +/- 4.8), (206.8 +/- 99.5), (1532.0 +/- 54.6), (159.2 +/- 11.0), (1612.0 +/- 515.9) and (2781.2 +/- 479.0) pg/(10(6)cell.24 h). Extracellular bFGF expressions of SW1990, Capan-1, Aspc-1, MiaPaCa-2, Panc-1 and PCT-3 were (2.1 +/- 0.6), (10.3 +/- 1.5), (31.0 +/- 0.4), (4.3 +/- 1.2), (43.6 +/- 1.5) and (82.1 +/- 10.4) pg/(10(6)cell.24 h). Intracellular endostatin expressions of SW1990, Capan-1, Aspc-1, MiaPaCa-2, Panc-1 and PCT-3 were (0.2 +/- 0.0), (0.3 +/- 0.0), (4.7 +/- 0.1), (10.8 +/- 0.2), (31.9 +/- 11.7) and (5.4 +/- 0.1) ng/(10(6)cell.24 h). Extracellular endostatin expressions of SW1990, Capan-1, Aspc-1, MiaPaCa-2, Panc-1 and PCT-3 were (0.0 +/- 0.0), (1.6 +/- 0.0), (21.5 +/- 1.1), (40.8 +/- 0.4), (129.2 +/- 1.0) and (20.1 +/- 1.8) ng/(10(6)cell.24 h). Panc-1 enjoying stronger invasion and proliferation showed stronger expressions of VEGF, bFGF and endostatin. Intracellular expressions of bFGF was stronger than extracellular, extracellular expressions of VEGF and endostatin were stronger than intracellular. CONCLUSION: Expressions of angiogenic factors regulated by cancer cell played an important role in progression of pancreatic carcinoma.

[Relationship between changes of serum concentrations of antiangiogenic factors and disease progression in patients of pancreatic carcinoma].

OBJECTIVE: To investigate the relationship between TSP-1, Angiostatin and Endostatin serum concentrations and progression of pancreatic adenocarcinoma. METHODS: Fifty-six patients with suspected pancreatic cancer were enrolled in the study and divided into resectable group (n = 32) and unresectable group (n = 24) according to evaluation and staging with dual phase helical CT. Histopathologic examinations included postoperative final pathology and preoperative fine needle biopsies. Peripheral blood concentrations of antiangiogenic factors Angiostatin, Endostatin and TSP-1 were detected by using ELISA methods, selecting samples of health people as a control. RESULTS: Serum concentrations of antiangiogenic factors in pancreatic cancer group were significantly higher than those in health group (P < 0.01). Serum concentrations of Endostatin, Angiostatin and TSP-1 were significantly increased in unresectable group, and highly expressed in patients whom tumor sizes were greater than 2 cm and tumor invaded peripancreatic great vessels (P < 0.05). After operation, serum concentrations of Endostatin, Angiostatin and TSP-1 significantly decreased (P < 0.05). There were no significant difference between I, II stage group and III, IV group. CONCLUSIONS: Detection of serum concentrations of antiangiogenic factors may be used to evaluate the resectability of pancreatic cancer and may play important roles in growth, invasion and metastasis of pancreatic cancer.

Diagnosis and management of isolated pancreatic tuberculosis: experience of 13 cases.

OBJECTIVE: To analyze the diagnosis and treatment of pancreatic tuberculosis. METHODS: Retrospectively reviewed and summarized 13 pancreatic tuberculosis patients' clinical information, presentation, diagnostic methods, therapeutic approaches, and prognosis from 1958 to 2004 at Peking Union Medical College Hospital. RESULTS: All cases presented a wide series of symptoms, including fever in 6 cases, upper abdominal tenderness in 13, epigastric mass in 4, obstructive jaundice in 3, night sweat in 4, weight loss in 7, hypersplenotrophy and hypersplenism in 1, and being complicated with tuberculosis of other organs in 3. One case was diagnosed by clinical symptoms and biopsy of lymph node, and only received anti-tubercular treatment Others were diagnosed by intra-operative biopsy and anti-tubercular treatment, and got well without recurrent tuberculosis in pancreas and other organs during 6 months to 2 years of follow-up. The non-operative case presented extrahepatic portal hypertension. CONCLUSIONS: Pancreatic tuberculosis may be considered in the patients with fever, abdominal tenderness, weight loss, and imaging evidence of regional pancreatic lesion. Efficacy of anti-tubercular agents and laparotomy for pancreatic tuberculosis is evident.