[Role of prostaglandin E2 in the modulation of renal water transport].

Prostaglandin E2 (PGE2), a bioactive lipid mediator, is one of the most important locally acting factors involved in a variety of physiological and pathophysiological processes. PGE2 binds with four EP receptors (EP1-4) to activate G protein-coupled receptor signaling responses. Recent functional and molecular studies have revealed that PGE2 plays an essential role in regulation of renal fluid transport via a variety of mechanisms. The water balance mainly depends on the regulation of aquaporin-2 (AQP2) by arginine vasopressin (AVP) in renal collecting duct principal cells. In recent years, increasing evidence suggests that PGE2 plays an important role in renal water reabsorption in the collecting ducts. In this paper, we reviewed the role of PGE2 and its receptors in the regulation of water reabsorption in the kidney, which may provide a new therapeutic strategy for many diseases especially nephrogenic diabetes insipidus.

Farnesoid X receptor (FXR) inhibits coagulation process via inducing hepatic antithrombin III expression in mice.

Farnesoid X receptor (FXR) has been identified as an inhibitor of platelet function and an inducer of fibrinogen protein complex. However, the regulatory mechanism of FXR in hemostatic system remains incompletely understood. In this study, we aimed to investigate the functions of FXR in regulating antithrombin III (AT III). C57BL/6 mice and FXR knockout (FXR KO) mice were treated with or without GW4064 (30 mg/kg per day). FXR activation significantly prolonged prothrombin time (PT) and activated partial thromboplastin time (APTT), lowered activity of activated factor X (FXa) and concentrations of thrombin-antithrombin complex (TAT) and activated factor II (FIIa), and increased level of AT III, whereas all of these effects were markedly reversed in FXR KO mice. In vivo, hepatic AT III mRNA and protein expression levels were up-regulated in wild-type mice after FXR activation, but down-regulated in FXR KO mice. In vitro study showed that FXR activation induced, while FXR knockdown inhibited, AT III expression in mouse primary hepatocytes. The luciferase assay and ChIP assay revealed that FXR can bind to the promoter region of AT III gene where FXR activation increased AT III transcription. These results suggest FXR activation inhibits coagulation process via inducing hepatic AT III expression in mice. The present study reveals a new role of FXR in hemostatic homeostasis and indicates that FXR might act as a potential therapeutic target for diseases related to hypercoagulation.

The effect of miRNA-122 in regulating fat deposition in a cell line model.

Accumulating evidence supports the role of miR-122 in fatty liver disease. We investigated miR-122 expression in a steatotic hepatocyte model, the effect of miR-122 over-expression and inhibition in the pathogenesis. Human hepatic cell line L02 was induced with oleic acid to establish the steatotic hepatocyte model. Intracellular lipid content was observed with laser scanning confocal microscope (LSCM), and triglyceride content was determined with kits. Total RNA was extracted and reversely transcribed into cDNA. miR-122 expression was measured using qRT-PCR. Subsequently, miR-122 mimic and miR-122 inhibitor were transfected into steatotic hepatocytes to observe their effect on intracellular lipid content. The lipid fluorescence intensity and triglyceride content within the steatotic hepatocytes were significantly higher than those in normal control (860.01 ± 26.52 vs. 257.77 ± 29.69 and 3.47 ± 0.12 vs. 1.85 ± 0.02 at 24 h) (P < 0.01). miR-122 expression in steatotic hepatocytes was down-regulated compared with that in control (2-ΔCt value: 0.0286 ± 0.0078 vs. 0.0075 ± 0.0012) (P ≪ 0.01). After transfection, miR-122 expression (2-ΔCt value) in the miR-122 mimic group increased 2.96-fold compared with that in control, and its lipid fluorescence intensity was significantly lower than that in control (790.92 ± 46.72 vs. 1,022.16 ± 49.66) (P < 0.01). Nevertheless, miR-122 expression decreased 3.45-fold in the miR-122 inhibitor group compared with that in control, and its fluorescence intensity was significantly higher than that in control (1,386.49 ± 40.34 vs 1,022.16 ± 49.66)(P ≪ 0.01). We concluded that miR-122 was down-regulated in steatotic hepatocytes model. The pathogenesis of hepatocyte steatosis was enhanced by miR-122 mimic and reduced with miR-122 inhibitor.

[Application of Modified DTT Methods to Reduce the Interference of Daratumumab on Serological Detection].

OBJECTIVE: To modified the classic dithiothreitol (DTT) method for treating red blood cells (RBCs) in Technical Manual of American Association of Blood Banks（AABB） and evaluate its application value in pre-transfusion examination of patients treated with daratumumab. METHODS: The classic 0.2 mol/L DTT method was improved in terms of PBS, DTT concentration, donor RBCs concentration (suspended/packed) and sample processing time. The modified DTT methods and AABB classic DTT method were applied to the blood matching tests of 12 multiple myeloma patients treated with daratumumab. The effect of treating panel RBCs with modified DTT methods on the detection of other irregular antibodies was evaluated by using antiserum and antibody reagents with known antibody properties. RESULTS: Two modified DTT methods were established (method 1: changed the concentration of DTT to 0.01 mol/L; method 2: changed the concentration of DTT to 0.02 mol/L and replaced the packed RBCs with 3% RBCs suspension). The optimal treatment time was 35 min for the modified DTT methods. At this time, the pan-agglutination caused by daratumumab was eliminated, but the detection of antibodies such as anti-E, anti-JKa, anti-M were not affected, and the titer of anti-K antibodies was only slightly decreased. CONCLUSION: The modified DTT methods were effective, which can eliminate the interference of daratumumab while retaining the activity of the Kell blood group system, and can replace the current classic DTT method in AABB Technical Manual.

The C-681G polymorphism of the PPAR-γ gene is associated with susceptibility to non-alcoholic fatty liver disease.

Non-alcoholic fatty liver disease (NAFLD) is defined as excessive accumulation of fatty acid in the liver, a common disease in the world. The research of single nucleotide polymorphisms (SNPs) provides a new approach for managing NAFLD. SNPs may increase or decrease the functions of the target genes and their encoding proteins. Peroxisome proliferator-activated receptor (PPAR) plays a key role in modulating metabolism of hepatic triglycerides and consequently magnitude of NAFLD. In this study, we investigated the effect of three SNPs in the PPAR-γ gene i.e. rs10865710 (C-681G), rs7649970 (C-689T) and rs1801282 (C34G, also termed Pro12Ala) on susceptibility to NAFLD. The participants were selected from our epidemiological survey. Totally 169 participants were enrolled in NAFLD group, and 699 healthy subjects were included as controls. PCR-RFLP was applied to detect the SNPs. The G allele frequency of rs10865710 in NAFLD group (41.1%) was significantly higher than that (34.8%) in controls (p = 0.03). Differences in other two loci (rs7649970 and rs1801282) were not statistically significant between the two groups (p > 0.05). This result was confirmed by haplotype analysis. The GCC haplotype (a set of 3 adjacent SNPs in linkage disequilibrium, corresponding to the three alleles of above polymorphisms in order) was a risk factor for the susceptibility to NAFLD (p = 0.03). This study has revealed that the G allele of rs10865710 in the PPAR-γ gene is associated with the increased susceptibility to NAFLD. Our findings may provide novel diagnostic biomarkers and therapeutic targets for NAFLD.

[Annual Nitrogen Removal Efficiency and Change in Abundance of Nitrogen Cycling Microorganisms in Swine Wastewater Treated by Crop Straw Materials].

Based on previous research, using straw material to treat swine wastewater can effectively reduce the concentration of nitrogen (N); however, the annual N-removal efficiency and change in the abundance of N-cycling functional genes remain unclear. In this study, four treatments (wheat straw, rice straw, corn stalk, and CK) were set up, with the aim of studying the annual N-removal efficiency and change in the abundance of functional genes. Our results showed that:① the total nitrogen (TN) removal and NH4+-N removal efficiency were the best in the first six months and were significantly reduced in the following six months. In addition, the TN removal and NH4+-N efficiency in straw and wheat straw were better than those in corn straw. The TN-removal efficiency in straw and wheat straw were 32.81%±11.34% and 32.99%±9.60%, respectively. The NH4+-N removal efficiency in straw and wheat straw were 35.3%±13.23% and 34.97%±12.00%, respectively. ② The abundance of N-cycling functional genes significantly increased by the addition of straw materials, compared with that of the CK (P<0.05). The average abundances of nirK, nirS, and hzsB were 6.45×109 copies·L-1, 6.18×109 copies·L-1, and 2.31×109 copies·L-1, respectively. The average abundances of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) were 6.12×1010 copies·L-1 and 4.93×109 copies·L-1, respectively. The average hzsB gene abundance was 2.31×109 copies·L-1. The average abundance of 16S rRNA in the treatment was 8.90×1010 copies·L-1. The abundances of hzsB and nirS genes in the straw and wheat straw were higher than those in the other treatment, indicating that the activities of anaerobic ammonia oxidation and denitrifying microorganisms were significantly increased by the addition of straw and wheat straw (P<0.05). In addition, the abundance of AOA and AOB genes were increased in wheat straw, suggesting that wheat straw could promote nitrification. The results provided data supporting the molecular mechanism of nitrogen removal in swine wastewater treatment with straw materials.

[General characteristics and clinical practices of Chinese patients with non-alcoholic fatty liver disease].

OBJECTIVE: To assess the characteristics and daily treatment compliance of non-alcoholic fatty liver disease (NAFLD) patients in China. METHODS: NAFLD adult patients from 21 clinics of 12 cities in China were enrolled in this registry. Physical examination such as demographic characteristics (height, weight, waist circumference measurement), blood pressure and clinical laboratory and ultrasonographic examination of liver were undertaken. Daily practice including life style and medication were recorded and assessed in accordance with 2006 Chinese NAFLD treatment guidelines. RESULTS: A total of 1656 patients were enrolled (1146 male and 510 female), mean of 45.8 ± 12.6 years old, mean duration of NAFLD history was (47.2 ± 47.7) months. 44.9% of NAFLD were suffering from metabolic syndromes. Patients with central obesity have higher incidence of hypertension and lower level of high-density lipoprotein cholesterol (HDL-C) than those without central obesity, P < 0.05. Body mass index (BMI), waist circumference, triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C) in ALT abnormal group were higher than those in ALT normal group (P < 0.05), HDL-C was lower in ALT abnormal group (P < 0.05). Significant differences existed between the BMI, female waist circumference, TG, fast insulin, HOMA index, ALT, AST and HDL-C among subgroups with mild, moderate and severe steatosis. Majority of the patients did not follow recommendations of NAFLD treatment guidelines. Among targeted population only 15.3% of patients used insulin sensitizers and 23.8% took lipid lowering medicine according to the guideline. CONCLUSION: Data indicated that nearly half of NAFLD patients co-morbid with metabolic disorders. Therapy compliance was unsatisfactory and the gap between current practice and Chinese NAFLD treatment guidelines was not optimal.

[Nitrogen and Phosphorus Removal in Surface Flow Constructed Wetland Planted with Myriophyllum elatinoides Treating Swine Wastewater in Subtropical Central China].

The loss of nitrogen (N) and phosphorus (P) from aquaculture has caused eutrophication of freshwater systems. Here, surface flow constructed wetland (SFCW) planted with Myriophyllum elatinoides were used to treat swine wastewater from a medium-sized hoggery in subtropical Central China. Inflow concentrations of NH4+-N, TN, TP, and COD ranged from 535.4 to 591.09, 682.09 to 766.96, 57.73 to 82.29, and 918.4 to 1940.43 mg·L-1, respectively. The mean removal efficiencies of NH4+-N, TN, TP, and COD were 97.4%, 97.1%, 91.0%, and 90.2%, respectively, and CW1 had the largest contributions of 37.3%, 38.4%, 43.3%, and 27.4%, respectively. Plant N and P uptake ranged 23.87-79.96 g·m-2 and 5.34-18.98 g·m-2, accounting for 19.1% and 20.2% of removal, respectively. Sediment N and P accumulation ranged 19.17-56.62 g·m-2 and 10.59-26.62 g·m-2, accounting for 19.8% and 61.7% of removal, respectively. Multiple linear regression showed that environmental factors explained 79.9% of the N removal and 70.1% of the P removal; DO was the main factor affecting N removal, and sediment adsorption was the key process in P removal. These results show that M. elatinoides constructed wetland can efficiently treat swine wastewater, thereby reduce the discharge of pollutants downstream.

Influence of polygenetic polymorphisms on the susceptibility to non-alcoholic fatty liver disease of Chinese people.

BACKGROUND AND AIM: The aim of this study was to investigate the influence of polygenetic polymorphisms, which play a role in the pathogenesis of metabolic syndrome, on the susceptibility to non-alcoholic fatty liver disease (NAFLD) of Chinese people. METHODS: The subjects were selected from an epidemiological survey in the Guangdong province of southern China. In each polymorphism study, 50-117 subjects who met the diagnostic criteria of NAFLD and had typical clinical and ultrasonographic findings were placed into the case group. Using a nested case-control design, the same numbers of matched people without NAFLD were included as controls. Single nucleotide polymorphisms (SNP) at nine positions in seven candidate genes were tested. These SNP were found to be associated with the pathogenesis of metabolic syndrome. Genetic analyses were performed using genomic DNA extracted from peripheral blood leukocytes. Polymerase chain reaction-restriction fragment length polymorphism was applied to detect SNP. RESULTS: Most candidate genes' SNP were associated with susceptibility to NAFLD. Some showed positive relationships (increased risk): tumor necrosis factor-alpha-238, adiponectin-45, leptin-2548, peroxisome proliferator-activated receptors-161 and phosphatidyletha-nolamine N-methyltransferase-175. Other SNP demonstrated a negative association (decreased risk): adiponectin-276 and hepatic lipase-514. Only two were not associated: tumor necrosis factor-alpha-380 and peroxisome proliferator-activated receptors-gamma co-activator-1alpha-482. CONCLUSION: Most candidate genes' SNP examined in metabolic syndrome patients were associated with susceptibility to NAFLD.

Experimental ablation of the pancreas with high intensity focused ultrasound (HIFU) in a porcine model.

The aim of this study was to determine the feasibility and safety of high intensity focused ultrasound's (HIFU) in pancreatic diseases. Twelve pigs were divided into three groups. The pancreases of pigs in Group A were ablated directly with HIFU, but those in Group B and C ablated by extracorporeal HIFU. The pigs in Group C were sacrificed at day 7 after HIFU. Serological parameters were determined pre-operation and post-operation. The entire pancreas was removed for histological examination. Each animal tolerate the HIFU ablation well. The complete necrosis was observed in targeted regions. The margins of the necrotic regions were clearly delineated from the surrounding normal tissues. Infiltration of inflammatory cells and phorocytosis on the boundary were found in group C. Blood and urine amylase levels were relatively steady after HIFU. No acute pancreatitis or severe complications occurred. In conclusion, HIFU ablation on the pancreas was safe and effective in experimental pigs.

[In vitro anti-tumor effect of cytotoxic T lymphocyte activated by antigen- loaded dendritic cells from peripheral blood mononuclear cells treated with calcium ionophore A23187 and GM-CSF].

OBJECTIVE: To evaluate the effect of calcium ionophore (CI) A23187 and human recombinant granulocyte/macrophage colony stimulating factor (rhGM-CSF) on the cultivation of dendritic cell (DC) from healthy human peripheral blood mononuclear cell (PBMC) and to evaluate the in vitro effect of DC stimulated by K562 cell lysate on inducing specific cytotoxic T lymphocyte (CTL) against K562 cell. METHODS: Human PBMCs isolated from healthy subjects were separated into two groups. In Group A, the cells were cultured with additional rhGM-CSF, recombinant human interleukin 4 and recombinant human tumor necrosis factor-α only as control group. In Group B, the cells were cultured in the presence of rhGM-CSF and CI A23187. The cells in both groups were pre-incubated with K562 cell lysate at 37°C for 30 min. The cells were harvested after a 4-day cultivation. Morphology of DC was continuously observed under inverted microscope. The surface antigens of induced cells were analyzed by flow cytometry (FCM). Then the proliferation of allogeneic T cell and the specific cytotoxicity of T cell primed with DC were examined by colorimetry. Also, the nonspecific inhibition of DC loaded K562 cell lysate against K562 cell was detected. RESULTS: Typical morphological features of DC could be observed in both groups. The expressions of CD83, CD1a, CD86 and CD40 were stronger in Group B than those in control group (45.2% ± 1.8%, 31.5% ± 3.9%, 40.1% ± 7.8%, 36.4% ± 6.3% vs 16.9% ± 1.3%, 20.4% ± 3.4%, 26.5% ± 2.2%, 22.3% ± 3.0%) (all P < 0.05). The expression of CD14 was weaker in Group B than that in control group (5.7% ± 0.8% vs 19.0% ± 1.6%) (P < 0.05). As compared with the control group, DC in Group B loaded with K562 lysate could evidently stimulate the proliferation of allogeneic T cell (P < 0.05, exclusion of effector-to-target ratio of 1:40) and inhibit the growth of K562 cell (P < 0.05). In addition, both groups of DC-stimulated CTL had specific cytotoxicity against K562 cell. At the effector-to-target ratios of 10:1 and 40:1, the DC-stimulated CTL of Group B had stronger cytotoxicity against K562 cell (both P < 0.05). CONCLUSION: In combination with rhGM-CSF, CI A23187 induces PBMC into DC in a more effective way. DC loaded with K562 lysate can stimulate CTL and maintain high immunocompetence with specific cytotoxicity against K562 cell.

The roles and interaction of FXR and PPARs in the pathogenesis of nonalcoholic fatty liver disease.

BACKGROUND AND STUDY AIMS: To identify the roles and interaction of farnesoid X receptor (FXR) and peroxisome proliferator activated receptors (PPARs) in Non-alcoholic fatty liver disease (NAFLD) pathogenesis. MATERIAL AND METHODS: 16 C57/BL male FXR knockout (KO) mice and sex- and age-matched C57/BL wild type mice were received either standard rodent chow or high-fat and sucrose diet (Blank control, NAFLD, FXR KO and FXR KO NAFLD) for 8 weeks. After that, all mice were sacrificed. Liver tissues and blood samples were collected for laboratory and RT-PCR examination. RESULTS: NAFLD, FXR KO and FXR KO NAFLD mouse models were successful established. Compared with blank control, FXR and PPAR-α mRNA expression decreased significantly (P < 0.05), PPAR-ß expression increased slightly (P > 0.05), PPAR-γ expression increased significantly in NAFLD (P < 0.05). Slight increased PPAR-α mRNA expression (P > 0.05) and markedly decreased PPAR-ß and PPAR-γ expression (P < 0.05) were found in FXR KO. Compared with FXR KO group, there was a slight increase in PPAR-αand PPAR-ßmRNA expression (P > 0.05) and significant increase in PPAR-γ expression (P < 0.05) in FXR KO NAFLD group. Comparison with NAFLD, PPAR-α mRNA expression increased slightly (P > 0.05), PPAR-ß and PPAR-γ expression decreased significantly (P < 0.05) in FXR KO NAFLD. CONCLUSION: FXR and PPARs interaction may play important roles in NAFLD pathogenesis.

Neuropsychological assessment of hepatic encephalopathy: ISHEN practice guidelines.

Low-grade or minimal hepatic encephalopathy (MHE) is characterised by relatively mild neurocognitive impairments, and occurs in a substantial percentage of patients with liver disease. The presence of MHE is associated with a significant compromise of quality of life, is predictive of the onset of overt hepatic encephalopathy and is associated with a poorer prognosis for outcome. Early identification and treatment of MHE can improve quality of life and may prevent the onset of overt encephalopathy, but to date, there has been little agreement regarding the optimum method for detecting MHE. The International Society on Hepatic Encephalopathy and Nitrogen Metabolism convened a group of experts for the purpose of reviewing available data and making recommendations for a standardised approach for neuropsychological assessment of patients with liver disease who are at risk of MHE. Specific recommendations are presented, along with a proposed methodology for further refining these assessment procedures through prospective research.

[Expression of NK cells receptor NKG2D from peripheral blood in patients with primary hepatic carcinoma and its clinical significance].

OBJECTIVE: To investigate the expression of NK cells receptor NKG2D from peripheral blood in patients with primary hepatic carcinoma and the relationship between NKG2D expression and cytotoxicity of NK cells. METHODS: Flow cytometry was used to determine the number of NK cells and the expression of NK cells receptor NKG2D from peripheral blood in patients with primary hepatic carcinoma (20 cases), hepatitis B cirrhosis (23 cases), hepatitis B (20 cases) and healthy control (20 cases). The microplate reader was used to detect cytotoxicity of NK cells in all cases. RESULTS: Among killing rate of NK cell for K562 cell, the expression rate of NKG2D in NK cells, the number of NKG2D(+)NK cells, NKG2D expression level of NK cells and the number of NK cells, the liver cancer group [(25 +/- 7)%, 6%, 0.7 x 10(7)/L, 15, (1.1 +/- 0.6) x 10(8)/L] decreased significantly as compared with the healthy group [(63 +/- 7)%, 36%, 8.3 x 10(7)/L, 116, (2.7 +/- 1.1) x 10(8)/L] and the hepatitis B group [(41 +/- 8)%, 16%, 2.8 x 10(7)/L, 49, (1.9 +/- 1.1) x 10(8)/L] (P < 0.05); and there was a slight decrease as compared with the hepatitis B cirrhosis group [(29 +/- 10)%, 7%, 0.6 x 10(7)/L, 29, (1.5 +/- 1.2) x 10(8)/L] (all P > 0.05 except NKG2D expression level of NK cells P < 0.05). The activity of NK cells showed a obvious positive correlation with the number of NK cell and the positive rate of NKG2D in NK cells, the number of NKG2D(+)NK cells and NKG2D expression level of NK cells (r = 0.657, 0.770, 0.927, 0.734, all P < 0.01). CONCLUSION: The cytotoxicity and the NKG2D expression of NK cells decreased significantly from peripheral blood in patients with primary hepatic carcinoma. The activity of NK cells was closely related to the NKG2D expression level of NK cells. Enhancing the NKG2D expression level of NK cell may provide a new idea for adoptive immunotherapy of primary hepatic carcinoma.

[Relationship between symptoms of pancreatic cancer-related depression and quality of life of patients].

OBJECTIVE: To investigate the relationship between symptoms of pancreatic cancer-related depression and quality of life of patients. METHODS: Fifty inpatients with pancreatic cancer from 3 Guangzhou hospitals between June 2007 and October 2008 were enrolled. Hamilton rating scale for depression-24 (HAMD-24) questionnaire was used to assess the degree of depression. Quality of Life (QoL) was evaluated by European Organization for Research and Treatment of Cancer Quality of Life Questionnaire-30 (EORTC QLQ-C30) and QLQ-PAN-26 respectively. RESULTS: Thirty-nine (78.0%) of these patients reported depression and 12 patients (30.8%) had severe depression. The incidence of depression in pancreatic cancer patients with chemotherapy was 92.3% (24/26), which was significantly higher than that of patients with surgical therapy (62.5%, 15/24) (P = 0. 011). The QoL of pancreatic cancer patients with depression in role functioning, emotional functioning and social functioning was significantly worse than that of patients without depression. The symptoms of fatigue, pain and appetite loss in pancreatic cancer patients with depression were significantly more than those without depression (P < 0.05). CONCLUSIONS: Depressive symptoms are common psychological disturbance in pancreatic cancer patients. Moreover, depression significantly lowers quality of life for pancreatic cancer patients.

[Decreased macrophage migration inhibitor factor expression up-regulats p27 in hepatocellular carcinoma.].

OBJECTIVES: To observe the expression of macrophage migration inhibition factor (MIF) and p27 in hepatocellular carcinoma tissue, and to investigate the effect of MIF on the expression of p27 in hepatocellular carcinoma (HCC) cells. METHODS: Immunohistochemistry and quantitative RT-PCR were performed to detect the expression of MIF and p27 in HCC tissues and peri-tumor tissues. Specific small interfering RNA (siRNA) targeting MIF gene was chemically synthesized and then transfected at the concentration of 50 nmol/L and 100 nmol/L into PLC cells and Hep3B cells. The mRNA levels of MIF and p27 after MIF siRNA treatment were quantified by real-time RT-PCR. RESULTS: MIF protein and mRNA were over-expressed in the HCC tumor tissues compared to these in the peri-tumor tissues (P less than 0.01). The expression of p27 protein and mRNA was significantly lower in the HCC tumor tissues compared to these in the peri-tumor tissues (P less than 0.01). Compared to normal liver cell line L-02, HCC cell lines expressed higher level of MIF (F=61.036, P less than 0.01) and lower level of p27 (F=529.853, P less than 0.01). In MIF siRNA treated PLC and Hep3B cells, the MIF mRNA was decreased in a dose-dependent manner (F=320.1, P less than 0.01; F=201.2, P less than 0.01). The p27 mRNA was significantly up-regulated in MIF siRNA treated PLC and Hep3B cells compared to control siRNA transfected cells (F=419.4, P less than 0.01; F=459.9, P less than 0.01). CONCLUSIONS: MIF is over-expressed in HCC tumor tissues, and the expression of p27 is repressed by MIF.

[Interference of osteopontin expression inhibits the invasion and metastasis of human hepatocellular carcinoma cell lines].

OBJECTIVE: To investigate the effect of osteopontin (OPN) on the invasion and metastasis of human hapatocellular carcinoma (HCC). METHODS: HCC cell lines (HCC-LM3) were transfected with the chemically synthesized small interfering RNA (siRNA). Real-time PCR and Western blot were used to quantify the mRNA and OPN protein levels. The malignant phenotypes including cellular growth, colony formation and invasion capability of the HCC cells were analyzed. RESULTS: The OPN mRNA and proteins levels were decreased by 75% and 80% in OPN siRNA treated cells. Colony formation and migratory capability were reduced in OPN siRNA treated cells (P < 0.05). CONCLUSION: The specific siRNA is able to reduce the OPN expression at both the mRNA and protein levels and significantly inhibits the invasiveness of HCC cells.

[Treatment of Highly Concentrated Swine Wastewater and Its Degradation Processes Using Three Matrix Materials].

The direct discharge of wastewater from pig farms can restrict wetland plant growth meaning that constructed wetlands (CWs) have generally low treatment efficiency. The treatment of farming wastewater using pre-ecological treatment technologies can be used to ensure that effluent concentrations reach the tolerable limits of wetland plants. This study focused on the efficient use of crop straw for reducing the pollution load of swine wastewater. Using field-scale pilot tests, wheat straw, straw, and corn stalk were used as test biological matrix pool fillers to treat the farming wastewater. Continuous intake of wastewater and a hydraulic retention time of 7 days was adopted. When the average concentrations of COD, TN, NH4+-N, NO3--N, and TP in the influent were 1652.83 mg·L-1, 371.31 mg·L-1, 303.51 mg·L-1, 0.67 mg·L-1, and 65.22 mg·L-1, respectively, wheat straw had the greatest removal effect on COD, TN, and TP, achieving a removal rate of 32.1%, 40.9%, and 33.3%, respectively. The removal efficiency of straw on NH4+-N was highest, reaching 43.4%. After 180 days, the lignin, cellulose, and hemicellulose of the three matrix materials were not completely decomposed. The degradation rate of lignin was lower than for cellulose and hemicellulose; the degradation of lignin and cellulose in the straw was fastest; and the degradation hemicellulose in wheat straw was fastest. The results show that wheat straw and straw offer a higher efficiency treatment for swine wastewater than corn stalk, and the suggested replacement cycle period is five months. These results provide initial data to support the application of biological matrix materials in the treatment of swine wastewater.

Effect of antidepressants on body weight, ethology and tumor growth of human pancreatic carcinoma xenografts in nude mice.

AIM: To investigate the effects of mirtazapine and fluoxetine, representatives of the noradrenergic and specific serotonergic antidepressant (NaSSA) and selective serotonin reuptake inhibitor (SSRI) antidepressant respectively, on body weight, ingestive behavior, locomotor activity and tumor growth of human pancreatic carcinoma xenografts in nude mice. METHODS: A subcutaneous xenograft model of human pancreatic cancer cell line SW1990 was established in nude mice. The tumor-bearing mice were randomly divided into mirtazapine group (10 mg/kg per day), fluoxetine group (10 mg/kg per day) and control group (an equivalent normal saline solution) (7 mice in each group). Doses of all drugs were administered orally, once a day for 42 d. Tumor volume and body weight were measured biweekly. Food intake was recorded once a week. Locomotor activity was detected weekly using an open field test (OFT). RESULTS: Compared to the fluoxetine, mirtazapine significantly increased food intake from d 14 to 42 and attenuated the rate of weight loss from d 28 to 42 (t = 4.38, P < 0.05). Compared to the control group, food intake was significantly suppressed from d 21 to 42 and weight loss was promoted from d 35 to 42 in the fluoxetine group (t = 2.52, P < 0.05). There was a significant difference in body weight of the mice after removal of tumors among the three groups. The body weight of mice was the heaviest (13.66 +/- 1.55 g) in the mirtazapine group and the lightest (11.39 +/- 1.45 g) in the fluoxetine group (F( (2,12) ) = 11.43, P < 0.01). The behavioral test on d 7 showed that the horizontal and vertical activities were significantly increased in the mirtazapine group compared with the fluoxetine and control groups (F( (2,18) ) = 10.89, P < 0.01). These effects disappeared in the mirtazapine and fluoxetine groups during 2-6 wk. The grooming activity was higher in the mirtazapine group than in the fluoxetine group (10.1 +/- 2.1 vs 7.1 +/- 1.9 ) (t = 2.40, P < 0.05) in the second week. There was no significant difference in tumor volume and tumor weight of the three groups. CONCLUSION: Mirtazapine and fluoxetine have no effect on the growth of pancreatic tumor. However, mirtazapine can significantly increase food intake and improve nutrition compared with fluoxetine in a pancreatic cancer mouse model.

[Investigation into the controlled release behavior of gemcitabine-loaded nanovesicles].

OBJECTIVE: To prepare gemcitabine-loaded nanovesicles and to observe its morphology, structure, particle size, and drug-release performance in vitro. METHODS: Diemulsion technique was used to prepare nanovesicles as carrier from amphiphilic block copolymer of poly (ethylene glycol)-block-poly (D, L-lactide), and gemcitabine was used as the model drug. The morphology of vesicles was determined by scanning electron microscope (SEM) and transmission electron microscope (TEM), and its drug loading (DL), encapsulation ratio (ER), and drug-release curve in vitro were detected by UV-Vis-NIR Spectrophotometer. RESULTS: The Gemcitabine-loaded nanovesicles is a kind of hollow nanosphere with the mean size of 200.6 nm, DL of 4.14% and ER of 20.54%. The nanovesicles showed its excellent controlled-release characteristic in the experiment of drug release in vitro. CONCLUSION: The nanovesicles prepared from PEG-PDLLA can be served as one of carriers for Gemcitabine with good performance of drug controlled-release. It will provide a reliable experimental base for the further researches in vivo.