The protective effects of rutaecarpine on acute pancreatitis.

Acute pancreatitis (AP) is the acute inflammation of the pancreas. The morbidity of AP has increased in recent years. Certain patients eventually develop severe AP (SAP), which rapidly progresses to multiple organ dysfunction; the incidence of this occurring in patients with AP is 20-30%. To date, no specific drugs or methods exist to treat this disease. Rutaecarpine relaxes vascular smooth muscle by stimulating calcitonin gene-related peptide (CGRP) release via activation of vanilloid receptor subtype 1 (VR1). It has been demonstrated that rutaecarpine induces a therapeutic effect on SAP. The present study was conducted to characterize the molecular mechanisms underlying the protective effects of rutaecarpine against AP using a rat model of AP. Gross pathological changes of the pancreas, as well as the pancreatic tissue histopathological score, were assessed following treatment with rutaecarpine, capsazepine or a combination of the two. Serum amylase activity was detected using an automatic biochemistry analyzer. Changes in the serum concentrations of interleukin (IL)-6, tumor necrosis factor (TNF-α), IL-10 and CGRP were assessed by ELISA and radioimmunoassay. The results demonstrated that pre-treatment with rutaecarpine markedly decreased pancreatic inflammation and necrosis, reduced the volume of ascites, and significantly increased the plasma concentration of CGRP and the serum concentration of IL-10, an anti-inflammatory cytokine. However, serum concentrations of the inflammatory cytokines IL-6 and TNF-α were decreased. The effect of rutaecarpine treatment markedly improved with increases in the drug dose. Capsazepine, as a competitive vanilloid receptor antagonist, abolished these protective effects of rutaecarpine against AP. Therefore, the results of the present study indicate that rutaecarpine protects against AP in rats by upregulating endogenous CGRP release via activation VR1 of, to improving the microcirculation of the pancreatic tissue and regulate the expression of inflammatory factors.

Changes in Histopathology, Enzyme Activities, and the Expression of Relevant Genes in Zebrafish (Danio rerio) Following Long-Term Exposure to Environmental Levels of Thallium.

Thallium is a rare-earth element, but widely distributed in water environments, posing a potential risk to our health. This study was designed to investigate the chronic effects of thallium based on physiological responses, gene expression, and changes in the activity of relevant enzymes in adult zebra fish exposed to thallium at low doses. The endpoints assessed include mRNA expression of metallothionein (MT)2 and heat shock protein HSP70; enzymatic activities of superoxide dismutase (SOD) and Na+/K+-ATPase; and the histopathology of gill, gonad, and liver tissues. The results showed significant increases in HSP70 mRNA expression following exposure to 100 ng/L thallium and in MT2 expression following exposure to 500 ng/L thallium. Significantly higher activities were observed for SOD in liver and Na+/K+-ATPase activity in gill in zebra fish exposed to thallium (20 and 100 ng/L, respectively) in comparison to control fish. Gill, liver, and gonad tissues displayed different degrees of damage. The overall results imply that thallium may cause toxicity to zebra fish at environmentally relevant aqueous concentrations.

Effects of triptolide on hippocampal microglial cells and astrocytes in the APP/PS1 double transgenic mouse model of Alzheimer's disease.

The principal pathology of Alzheimer's disease includes neuronal extracellular deposition of amyloid-beta peptides and formation of senile plaques, which in turn induce neuroinflammation in the brain. Triptolide, a natural extract from the vine-like herb Tripterygium wilfordii Hook F, has potent anti-inflammatory and immunosuppressive efficacy. Therefore, we determined if triptolide can inhibit activation and proliferation of microglial cells and astrocytes in the APP/PS1 double transgenic mouse model of Alzheimer's disease. We used 1 or 5 µg/kg/d triptolide to treat APP/PS1 double transgenic mice (aged 4-4.5 months) for 45 days. Unbiased stereology analysis found that triptolide dose-dependently reduced the total number of microglial cells, and transformed microglial cells into the resting state. Further, triptolide (5 µg/kg/d) also reduced the total number of hippocampal astrocytes. Our in vivo test results indicate that triptolide suppresses activation and proliferation of microglial cells and astrocytes in the hippocampus of APP/PS1 double transgenic mice with Alzheimer's disease.

Two novel compound heterozygous mutations associated with types I and II protein C deficiency with unusual phenotypes.

INTRODUCTION: We diagnosed two Chinese hereditary PC deficiency families and identified two novel compound heterozygous mutations (p.Arg194Cys/Gly324Ser and p.Glu274X/Asp297His) in the protein C (PROC) gene. The probands were classified as types I and II PC deficiency. The aim of this article is to access the influence of the mutations on PC activity, antigen and protein structure, and to evaluate whether there is abnormal PC localization. MATERIALS AND METHODS: Genomic DNA of all family members was extracted, PCR amplified, and sequenced. The mutant PC expression plasmids were constructed. Expression assays, intracellular localization, and molecular modeling were performed. RESULTS: Proband 1, a type II PC defect, harbored a compound heterozygous mutation, p.Arg194Cys/Gly324Ser in the PROC gene, underwent two thromboembolic events. Expression assays indicated that the p.Arg194Cys mutant lead to decreased PC activity and normal PC Ag levels. Intracellular localization showed that both p.Arg194Cys and p.Gly324Ser co-localized with the endoplasmic reticuli and the Golgi apparatus. Molecular modeling suggested that the p.Gly324Ser mutation disturbed the interaction between the heavy and light chains of the PC protein. Proband 2, a type I PC defect, harbored a compound heterozygous PROC gene mutation, p.Glu274X/Asp297His, presented with recurrent spontaneous abortion and right popliteal vein thrombosis. Expression results were in accordance with the PC changes of the patient, and existed in defective PC transport. Structural model suggested p.Glu274X lead to disulfide bond between heavy and light chain cannot form. CONCLUSIONS: Our results confirm that two novel compound heterozygous PROC gene mutations are causative on the two PC deficiency families.

Tumor-specific expression of shVEGF and suicide gene as a novel strategy for esophageal cancer therapy.

AIM: To develop a potent and safe gene therapy for esophageal cancer. METHODS: An expression vector carrying fusion suicide gene (yCDglyTK) and shRNA against vascular endothelial growth factor (VEGF) was constructed and delivered into EC9706 esophageal cancer cells by calcium phosphate nanoparticles (CPNP). To achieve tumor selectivity, expression of the fusion suicide gene was driven by a tumor-specific human telomerase reverse transcriptase (hTERT) promoter. The biologic properties and therapeutic efficiency of the vector, in the presence of prodrug 5-fluorocytosine (5-FC), were evaluated in vitro and in vivo. RESULTS: Both in vitro and in vivo testing showed that the expression vector was efficiently introduced by CPNP into tumor cells, leading to cellular expression of yCDglyTK and decreased VEGF level. With exposure to 5-FC, it exhibited strong anti-tumor effects against esophageal cancer. Combination of VEGF shRNA with the fusion suicide gene demonstrated strong anti-tumor activity. CONCLUSION: The shVEGF-hTERT-yCDglyTK/5-FC system provided a novel approach for esophageal cancer-targeted gene therapy.

Dissecting characteristics and dynamics of differentially expressed proteins during multistage carcinogenesis of human colorectal cancer.

AIM: To discover novel biomarkers for early diagnosis, prognosis or treatment of human colorectal cancer. METHODS: iTRAQ 2D LC-MS/MS analysis was used to identify differentially expressed proteins (DEPs) in the human colonic epithelial carcinogenic process using laser capture microdissection-purified colonic epithelial cells from normal colon, adenoma, carcinoma in situ and invasive carcinoma tissues. RESULTS: A total of 326 DEPs were identified, and four DEPs (DMBT1, S100A9, Galectin-10, and S100A8) with progressive alteration in the carcinogenic process were further validated by immunohistochemistry. The DEPs were involved in multiple biological processes including cell cycle, cell adhesion, translation, mRNA processing, and protein synthesis. Some of the DEPs involved in cellular process such as "translation" and "mRNA splicing" were progressively up-regulated, while some DEPs involved in other processes such as "metabolism" and "cell response to stress" was progressively down-regulated. Other proteins with up- or down-regulation at certain stages of carcinogenesis may play various roles at different stages of the colorectal carcinogenic process. CONCLUSION: These findings give insights into our understanding of the mechanisms of colorectal carcinogenesis and provide clues for further investigation of carcinogenesis and identification of biomarkers.

Cell motility and spreading promoted by CEACAM6 through cyclin D1/CDK4 in human pancreatic carcinoma.

Carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6) belongs to the human carcino-embryonic antigen (CEA) family. Numerous lines of studies have indicated that altered expression of CEACAM6 may have a role in carcinogenesis and development. However, few studies have defined functional roles and mechanisms of action. In the present study, the relationship between clinical and pathological parameters was also analyzed. The relative CEACAM6 protein expression of pancreatic carcinoma was significantly higher than that in non-cancerous tissue. Different clinical stages and lymph node metastasis between groups were significantly different (P<0.05). We used siRNA and forced-expression in multiple cell lines to define the role of CEACAM6 in the regulation of proliferation of pancreatic carcinoma in vitro and in vivo. Knockdown of endogenous CEACAM6 decreased proliferation of BxPC-3 and SW1990 cells. These changes significantly reduced cyclin D1 and CDK4 protein levels. Conversely, overexpression of CEACAM6 in MIA PaCa-2 cells stimulated proliferation and increased cyclin D1 and CDK4 protein levels. Our results confirm that CEACAM6 promoted cell proliferation, and these changes were mediated by cyclin D1/CDK4. These observations contribute to our understanding of the important roles of CEACAM6 in pancreatic carcinoma development and progression and could be a promising molecular target for the development of new diagnostic and therapeutic strategies of pancreatic carcinoma.

Silencing of miR-21 sensitizes CML CD34+ stem/progenitor cells to imatinib-induced apoptosis by blocking PI3K/AKT pathway.

BCR-ABL tyrosine kinase inhibitor imatinib fails to eradicate leukemia stem cells (LSCs), the underlying mechanisms maintaining CML LSCs remain poorly understood. Here, we showed that transient inhibition of miR-21 by antagomiR-21 markedly increased imatinib-induced apoptosis in CML, but not normal CD34+ stem/progenitor cells. Furthermore, PI3K inhibitors also significantly sensitized CML CD34+ cells to imatinib-induced apoptosis. MiR-21 or PI3K inhibitor in combination with imatinib treatment significantly decreased AKT phosphorylation and c-Myc expression than either agent did alone, but did not affect Bim and Bcl-6 expresssion. These findings indicate that miR-21 is required for maintaining the imatinib-resistant phenotype of CML CD34+ cells through PI3K/AKT signaling pathway, thus providing the basis for a promising therapeutic approach to eliminate CML LSCs.

[Analysis of composition characteristics of municipal solid waste in south China].

Using the MSW (municipal solid waste) sampling and analysis methods, the composition characteristics of MSW in south China were investigated. The results showed that: the average MSW bulk density was 0.22 x 10(3) kg x m(-3), the percentages of water, ash and combustible were 55.0%-66.9%, 18.6%-30.3% and 69.7%-81.4%, respectively. The organic contents were 50.1%-58.0%. The waste low caloric value (wet base) ranged 6570-9652 kJ x kg(-1), and the average of waste low caloric value(wet base) was 8272 kJ x kg(-1), which was higher than the recommended value of World Bank(7000 kJ x kg(-1)). The percentage composition of MSW was: 39.8%-53.3% kitchen waste garbage, 16.5%-33.4% rubber and plastics, 5.61%-7.95% paper, 1.14%-5.16% textile products, 2.49%-5.12% bamboo products, 1.10%-1.47% glass, 5.86%-7.57% mixed materials, 2.46%-6.73% dust, 0.1%-0.32% metal, and 0.4%-0.69% ceramic. The correlation analysis and factor analysis showed that: the high proportions of textile, rubber and plastics, paper, and combustible materials had a positive effect on the MSW incineration, while the high proportions of kitchen waste garbage, glass, MSW bulk density, ash, water content and dust had a negative effect on the MSW incineration.

[Compositions and distribution characteristics of polybrominated diphenyl ethers in serum of women from Sichuan province].

The serum concentrations of polybrominated diphenyl ethers (PBDEs) in women from Sichuan province were detected via gas chromatograph-mass spectrometry in selected ion monitoring mode (SIM). Based on the experimental data, the compositions and distribution characteristics of PBDEs as well as pollution resource were also explored. Among these 11 congeners of PBDEs detected, BDE-209 was the dominant congener accounting for 63.57% -90.34% in the total PBDEs, followed by BDE-66,-99 and-100. The concentrations of BDE-209 ranged from 0.12 to 2.38 microg x L(-1) and the mean concentration was 0.63 microg x L(-1). Other congeners of these 11 PBDEs could not be detected. The data indicated that the major pollution came from deca-BDE, followed by tetra-BDE and penta-BDE. At the mean time, this result also showed that the pollution of PBDEs in Sichuan province was still at a lower level for ordinary women when compared to the data from other countries.

Quantitative proteomic analysis identifying three annexins as lymph node metastasis-related proteins in lung adenocarcinoma.

Lymph node status is a strong predictor of outcome for lung adenocarcinoma (AdC) patients. To explore novel potential protein markers for predicting lymph node metastasis of lung AdC, differential proteomic analysis on microdissected cancer cells from primary lung AdC and matched lymph node (LN) metastatic tissues by laser capture microdissection (LCM) was conducted using two-dimensional differential in-gel electrophoresis (2D-DIGE) coupled with mass spectrometry (MS). Annexins including annexin-1, annexin-2 and annexin-3 were identified and found to be overexpressed in matched LN metastatic tissues compared to primary lung AdC. Furthermore, differential expression levels of the three annexins were evaluated in paraffin-embedded 188 primary lung AdC tissues and 65 matched positive lymph node specimens using immunohistochemistry. High expression of annexin-1, annexin-2, and annexin-3 was all frequently observed in matched positive lymph node tissues compared to primary lung AdC. In primary lung AdC, expression levels of the three annexins in primary lymph node-positive AdC tissues were higher than primary lymph node-negative AdC tissues. Multivariate logistic regression analysis indicated annexin-1, annexin-2, and annexin-3 were all significant risk factors for lymph node metastasis. Furthermore, statistical analysis indicated that the concomitant expression of annexin-1/annexin-2, annexin-1/annexin-3, or annexin-2/annexin-3 and combined expression of all three markers had stronger correlation with lymph node metastasis. Our results suggest that annexin-1, annexin-2, and annexin-3 are identified as potential biomarkers associated with lymph node metastasis in lung AdC.

[Study of symbolic element in roasting slag of pyrite by inductively coupled plasma mass spectrometry].

In the present paper, the correlations between sixteen elements from the dry method roasted dust-slag of pyrite and sixteen elements from the soil layer near and far from the store area were studied by ICP-MS. Similar radio and outstanding different radio of the elements between the dust-slag and the soil were studied too. It was discovered that in the pollution soil layer Tl, Cd, Cs, Cu, Zn, Mn, Pb and Ni were easy to be identified and had similar radio with the elements in the dust-slag. But only Tl, Cd, Zn and Ni are suitable for criterion of element similar properties. In dust-slag and soil, distinct composition element radios of Tl, Cs, Co, Mo, Zn, Cr, V, Sr, Sb, Pb, Rb, Mn and Ni had striking differentiation. Only Tl, V, Sb and Cu corresponded to both the uncorrelated elements analysis of surface layer and middle-base layer soil. Tl could be used as an inspection target of similar elements and outstanding different elements between the dust-slag and the soil in the meanwhile. So we suggested that Tl can be used as a symbolic element in the roasting dust-slag of pyrite to find the dust-slag of pyrite in dust-recognition and to differentiate the metallurgy dust of pyrite and soil dust.

[Effects of eight environmental endocrine disruptors on insulin resistance in patients with polycystic ovary syndrome: a preliminary investigation].

OBJECTIVE: To investigate the relationship between 8 endocrine-disrupting chemicals in the serum and insulin resistance in patients with polycystic ovary syndrome (PCOS). METHODS: This study was conducted among 60 patients with PCOS, including 23 with insulin resistance (PCOS-IR) and 37 without insulin resistance (PCOS-NIR), and 29 non-PCOS women seeking medical attention for infertility or menstrual disorder (control group). The serum levels of 6 phthalic acid esters (PEAs), bisphenol A (BPA) and octylphenol (OP) were measured in all the subjects. RESULTS: The levels of PAEs, BPA and OP showed no significant differences between PCOS patients and the control group (P>0.05). The serum level of OP was significantly lower in patients PCOS-IR than in those with PCOS-NIR (47.89 ng/ml vs 60.24 ng/ml, P<0.05). CONCLUSION: PEAs and BPA do not produce obvious effect on the pathogenesis of PCOS or contribute to insulin resistance, but OP may play a role in insulin resistance in PCOS patients.

[Distribution of heavy metals in waters and pollution assessment in thallium contaminated area of Yunfu, Guangdong].

Distribution of Thallium(T1), Cadmium( Cd), Chromium (Cr), Lead (Pb), Nickel (Ni), Arsenic (As), Copper (Cu), Zinc (Zn) in water and sediments of Yunfu pyrite mine area was studied. The environmental risk assessment was conducted systematically using Sediment Quality Guidelines (SQGs) and Hakanson potential ecological risk index. The results indicated that concentration range of Tl in stream water was 0.19-65.25 microg/L, which is higher than the environmental quality standards for surface water. Concentration ranges of Tl, Zn, As, Cd, Pb in sediments were 5.89-63.0 mg/kg, 1215-5754 mg/kg, 208.4-1327 mg/kg, 4.20-17.5 mg/kg, 282-13,770 mg/kg. According to Sediments Quality Guidelines, sediments was moderately to severe level of pollution since concentrations of Tl, Cd, Cr, Pb, Ni, As, Cu, Zn were much higher than LEL (lowest effect level) values, and the concentrations of Pb, As, Zn were higher than SEL (severe effect level) values, the others were partly higher than SEL values, which may result in severe negative biota effects in the watersheds. Compared to soil background values of Guangdong province, the metals in stream sediment showed strong to severe strong ecological risk, and the ecological risk of heavy metals in the descending order of Tl, Cd, As, Pb, Cr, Ni, Zn and Cu. Besides, the sediments were severe contained by toxic element thallium and cadmium. Besides, the mainly ecological risk of heavy metal is thallium. More emphasis should be placed on thallium and cadmium control and disposal in

Study on oxidation of pyrite by in situ absorption spectroscopy.

Pyrite is one of the common natural mineral. It is easily oxidized and brings heavy metal contaminations, therefore it is the main source of acidity mine drainage. The study on the dynamics of pyrite is helpful to comprehending the mechanism of its pollution. In the present paper, an experimental method was designed that phenanthroline reacts with Fe2+ that is released from pyrite in solution, and produces stable orange-red complex compound. It can be detected by absorption spectroscopy. In-situ characterization of oxidation of pyrite can be achieved by this method. The results showed that the method is reliable and accurate, and it has high sensitivity and little interference; the reaction rate of oxidation increased linearly with time, corresponding to the characterization of zero-order reaction; oxidation of pyrite belongs to the surface reaction and the process of surface reaction is rate determining step.

Proteomic analysis of the stroma-related proteins in nasopharyngeal carcinoma and normal nasopharyngeal epithelial tissues.

The stroma surrounding cancer cell population is increasingly recognized as playing an important role in cancer proliferation, invasion, and metastasis. To identify the stromal proteins involved in nasopharyngeal carcinoma (NPC) carcinogenesis, differences in protein expression of the stroma from NPC and normal nasopharyngeal epithelium tissues (NNET) were assessed using a comparative proteomic approach combined with laser capture microdissection (LCM). LCM was performed to purify stromal cells from NPC and NNET, respectively. Proteins between the pooled microdissected tumor and normal stroma were separated by two-dimensional electrophoresis (2-DE) and differential proteins were identified by mass spectrometry (MS). Sixty differential proteins between normal stroma (NS) and tumor stroma (TS) were identified, and the expression of CapG protein was further confirmed by western blotting and immunohistochemical analysis. Our results will be helpful to study the role of stroma in the NPC carcinogenesis and may provide helpful clues for pathogenesis, early diagnosis, and progression of NPC.

[A spectral analysis method for total volatile base nitrogen].

A new method for the determination of the total volatile basic nitrogen (TVB-N) by reflectance spectroscopy was developed. The method was based on the reaction of TVB-N with Nessler's reagent treated by molecular sieve and the reflectance-absorption value F(R), which is directly proportional to the amount of NH2 Hg IO in solid phase, was measured by the reflection spectrometer. The fundamental principle, effective factors and experimental conditions of the method were discussed. The special features of this method were its simplicity in operation, relatively high sensitivity of determination and the use of a small amount of reagent. Linear calibration graph was obtained in the range 1-8 microg x mL(-1) with a detection limit of 0.1 microg x mL(-1). The method was applied to monitor the fresh degree of fish and pork. The result discovered that the total volatile basic nitrogen increased rapidly along with the time extension, indicating that the deterioration of fish and pork was an acceleration process, and the deterioration speed of fish was faster than that of pork.

[Characterization of kale (Brassica oberacea var acephala) under thallium stress by in situ attenuated total reflection FTIR].

The experiment was designed based on consumption of carbon dioxide through the photosynthesis of Brassica oberacea var acephala leaf, and the photosynthesis of kale leaf under thallium stress was investigated by in situ attenuated total reflection FTIR (in situ ATR-FTIR). The ATR-FTIR showed that the absorption peaks of leaves had no obvious difference between plants growing in thallium stress soil and plants growing in non-thallium pollution soil, and the strong peaks at 3,380 cm(-1) could be assigned to the absorption of water, carbohydrate, protein or amide; the strong peaks at 2,916 and 2,850 cm(-1) assigned to the absorption of carbohydrate or aliphatic compound; the peaks at 1,640 cm(-1) assigned to the absorption of water. However, as detected by the in situ ATR-FTIR, the double peaks (negative peaks) at 2,360 and 2,340 cm(-1) that are assigned to the absorption of CO2 appeared and became high gradually. It was showed that kale was carrying photosynthesis. At the same time, the carbon dioxide consumption speed of leaf under thallium stress was obviously larger than that of the blank It was expressed that photosynthesis under thallium stress was stronger than the blank All these represented that kale had certain tolerance to the heavy metal thallium. Meanwhile, the carbon dioxide consumption of grown-up leaf was more than that of young leaf whether or not under thallium stress. It was also indicated that the intensity of photosynthesis in grown-up leaf is higher than that in young leaf.

Identificating 14-3-3 sigma as a lymph node metastasis-related protein in human lung squamous carcinoma.

In this study, we aim to screen metastasis-related proteins in human lung squamous carcinoma (LSC) using laser capture microdissection and a proteomic approach. Twenty two differential proteins were identified from pooled microdissected primary LSC and matched lymph node (LN) metastatic tissues. Expression of the differential protein 14-3-3 sigma was determined by Western blotting and immunohistochemistry. In cell invasion assay, down-regulated 14-3-3 sigma by siRNA increased in vitro invasive ability of HTB-182 and A549 cells, up-regulation of 14-3-3 sigma by pcDNA3.0/14-3-3 sigma decreased in vitro invasive ability of HTB-182 and A549 cells. The data suggest that 14-3-3 sigma is a potential LN metastasis-related protein in LSC, and its dysregulation might play an important role in the LN metastatic process of LSC.

Quantitative proteomic analysis of differential proteins in the stroma of nasopharyngeal carcinoma and normal nasopharyngeal epithelial tissue.

The importance of stromal cells and the factors that they expressed during cancer initiation and progression have been highlighted by recent literature. To identify the stromal proteins involved in nasopharyngeal carcinoma (NPC) carcinogenesis, we assessed differences in protein expression of the stroma from NPC and normal nasopharyngeal epithelium tissues (NNET) using a quantitative proteomic approach combined with laser capture microdissection (LCM). LCM was performed to purify stromal cells from the NPC and NNET, respectively. The differential proteins between the pooled microdissected tumor and normal stroma were analyzed by two-dimensional difference gel electrophoresis (2D-DIGE) combined with mass spectrometry (MS). Twenty differential proteins were identified, and the expression and location of two differential proteins (L-plastin and S100A9) were further confirmed by Western blotting and immunohistochemical analysis. Our results will be helpful to study the role of stroma in the NPC carcinogenesis, as well as discover the interaction between NPC cells and their surrounding microenvironment.