[SERPINE2 promotes cellular migration and invasion in esophageal squamous cell carcinoma by activating ß-catenin].

Objective: To clarify the function and molecular mechanisms of serpin family E member 2 (SERPINE2) in cellular migration and invasion of esophageal squamous cell carcinoma (ESCC). Methods: The expression of SERPINE2 in ESCC was analyzed by using online databases TCGA (http: //gepia.cancer-pku.cn/detail.php and http: //ualcan.path.uab. edu/index.html). The expressions of SERPINE2 mRNA in normal human esophageal epithelial cell line NE2, human ESCC cell lines KYSE30 and KYSE150 were detected by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). SERPINE2-konckdown or SERPINE2-overexpressed plasmid was transfected into KYSE30 cells, and the efficiencies of the knockdown and overexpression system were tested by qRT-PCR. The relationships of SERPINE2 and ESCC migration and invasion were determined by migration and invasion assays in vitro. The associations between SERPINE2 expression and ß-catenin as well as its target genes including c-Myc, cyclin D1 and CD44 were analyzed by immunofluorescence, qRT-PCR and western blot, respectively. Results: The expressions of SERPINE2 were significantly upregulated in both esophageal cancer (ESCA) and ESCC tissues compared to normal tissues by analyzing 182 and 95 cases, respectively (P<0.01). SERPINE2 is highly expressed in both KYSE30 and KYSE150 cells (P<0.05). The number of migrating and invading cells in control group were (212.66±24.11)/field and (136.00±14.42)/field, while were (88.33±9.71)/field and (77.00±9.53)/field in SERPINE2-knockdown 1 group, and (66.00±8.00)/field and (45.66±3.78)/field in SERPINE2-knockdown 2 group, respectively, and the differences were dramatically significant compared with the control group (P<0.01). The number of migrating and invading cells in control group were (250.00±30.00)/field and (203.33±15.27)/field, while were (383.33±35.11)/field and (246.66±25.16)/field in SERPINE2-overpressed group, and the differences were strikingly significant compared with the control group (P<0.01). The protein expression of ß-catenin was upregulated while phosphorylated ß-catenin protein expression was downregulated in SERPINE2-overexpressed KYSE30 cells when compared to control cells.The transcription activity of ß-catenin was significantly upregulated and the mRNA expressions of its target genes including c-Myc, cyclin D1 and CD44 were all increased. After treated with 25 µM iCRT14, the number of migrated cells in the control and SERPINE2-overpressed groups were (200.00±36.05)/field and (258.33±22.54)/field, and the number of invaded cells were (160.00±17.32)/field and (188.33±25.65)/field, respectively, the differences were dramatically significant compared with the group without iCRT14 treatment (P<0.01). Conclusion: SERPINE2 is significantly upregulated in ESCC cells and can promote cellular migration and invasion by activating ß-catenin, which may provide a potential therapeutic target for patients with ESCC.

A clinical characteristics and genetic analysis of a case of Rubinstein-Taybi syndrome with glaucoma.

The aim of this study was to analyze the clinical features of a Rubinstein-Taybi syndrome (RSTS) case with neonatal glaucoma. We also wanted to explore the manifestation of the disease in combination with genotype-phenotype correlation. For DNA extraction we used 2 ml peripheral blood, collected from the child and parents. The extracted genomic DNA was used for clinical exome sequencing. A 38-day old baby boy was diagnosed with congenital glaucoma on the third day after birth with symptoms, including choking milk, feeding difficulties and slow weight gain. He was admitted to the neonatology department because of lung infection. The clinical exome sequencing showed that the child has a c.2368C>T heterozygous mutation in exome 13 in CREBBP (cAMP responsive element binding protein) while his parents have no such mutation. Combining genetic data with the clinical features, this infant was diagnosed with RSTS. This is the first report of RSTS caused by a c. 2368C>T mutation in CREBBP. RSTS is an extremely rare disease with extensive clinical manifestations. It is highly overlapped with other syndromes which makes the diagnosis difficult. RSTS is easy to be missed or misdiagnosed due to the lack of specific clinical manifestations during the neonatal period. Neonatal specialists need to enhance their awareness and recognition of this condition, and use genetic testing as an effective tool in order to finalize their diagnosis.

Effects of in ovo feeding of carbohydrates and arginine on hatchability, body weight, energy metabolism and perinatal growth in duck embryos and neonates.

1. The objective of this study was to test the hypothesis that in ovo feeding of carbohydrates and arginine into the duck amnion may improve the glycogen store and perinatal growth. At 23 d of incubation, fertile eggs were injected with 1·2 ml of sodium chloride (NaCl), sucrose + maltose (CHO), arginine (Arg) or sucrose + maltose + arginine (CHO + Arg), with controls not injected. Body weight, liver and muscle glycogen levels, and hepatic glucose-6-phosphatase activity were determined at 25 d of incubation, at hatch, and at 3 and 7 d posthatch. 2. At hatch and 7 d of age, the body weights were greater in the in ovo-feeding treatments than the controls. Arg and CHO + Arg significantly enhanced liver glycogen level at hatch compared with controls. CHO and CHO + Arg significantly increased muscle glycogen level at 25 d of incubation over controls. CHO and Arg decreased glucose-6-phosphatase at 25 d of incubation, whereas NaCl and CHO + Arg increased glucose-6-phosphatase at hatch relative to controls. 3. In ovo feeding of carbohydrates and arginine at 23 d of incubation may improve glycogen reserves, which may, in turn, provide the energy needed for perinatal growth.

Expression of N-methyl-D-aspartate receptor and its effect on nitric oxide production of rat alveolar macrophages.

We early show that glutamate (Glu) mediate hyperoxia-induced newborn rat lung injury through N-methyl-D-aspartate receptor (NMDAR). In this study, we search for evidence of NMDAR expression on newborn rat alveolar macrophages (AMs) and the difference between newborn and adult rat AMs, and the possible effect on nitric oxide (NO) production of AMs by exogenous NMDA. The protein of NMDAR was showed by immunocytochemistry, and the mRNA was examined by RT-PCR and real-time PCR. The results show that: (i) both newborn and adult rat AMs express NMDAR1 and the four NMDAR2 subtypes and newborn rat AMs are higher expression. (ii) NMDA administration increase NO production, inducible nitric oxide synthase (iNOS) activity and iNOS mRNA expression of AMs. (iii) NMDAR activation elevates NO secretion of AMs, which suggests that AM may be one of the key cellular origin of the elevated NO secretion in hyperoxia-induced lung injury.

Effects of 2-hydroxy-4-methylthiobutyrate on portal plasma flow and net portal appearance of amino acids in piglets.

To determine whether portal plasma flow (PPF) and net portal appearance of amino acids (AA) could be affected by 2-hydroxy-4-methylthiobutyrate (HMB), six barrows (35-day-old, 8.6+/-1.4 kg), implanted with arterial, portal and mesenteric catheters, were fed a DL-methionine (as the control) or HMB-supplemented diet once hourly and infused intramesenterically with 1% p-amino hippurate. PPF was numerically 9% higher (P=0.09) in HMB-fed pigs than in controls over a 4-6 h period. Compared with controls, pigs fed the HMB diet had increased (P<0.05) net portal balance and/or appearance of leucine, isoleucine, histidine, arginine and alanine, but had decreased (P<0.05) portal appearance of glutamate over a 6-h period. The concentration of acetate in the lumen of the distal small intestine was higher (P=0.01) in HMB-fed pigs than in controls (25.14 vs. 7.64 mmol/kg). mRNA levels for proglucagon and endothelial nitric-oxide synthase (eNOS) in stomach and proximal small intestine, and mRNA levels for GLP-2 receptor (GLP-2R) in stomach were higher (P<0.05) in HMB-fed pigs compared with those in controls. Collectively, HMB supplementation increased concentrations of short-chain fatty acids in intestinal lumen, expression of proglucagon, GLP-2R, and eNOS genes, and net portal absorption of AA. These novel findings from the study with pigs may also have important implications for intestinal nutrition and health in humans.

Duration of feeding linseed diet influences expression of inflammation-related genes and growth performance of growing-finishing barrows.

The aim of the study was to investigate the effects of dietary linseed (rich in n-3 PUFA) on expression of inflammation-related genes and on growth performance of growing-finishing barrows. Two isoenergetic and isonitrogenous diets were formulated, one as the basal diet and the other containing 10% linseed. Twenty-four Landrace x Yorkshire barrows weighing 35 +/- 3.7 kg were randomly assigned to 1 of 4 treatment groups, with 6 pigs per group. During the entire experimental period of 90 d, these 4 groups of pigs were first fed the basal diet and then fed the linseed diet for 0, 30, 60, and 90 d before slaughter, respectively. Pig growth; messenger RNA (mRNA) expression of peroxisome proliferator-activated receptor-gamma (PPARgamma), IL-1beta0, IL-6, and tumor necrosis factor-alpha (TNF-alpha); and plasma concentrations of the 3 proinflammatory cytokines were measured and analyzed. Average daily feed intake did not differ among treatment groups (P > 0.05), but ADG (P < 0.05) and G:F (P < 0.01) responded quadratically to the duration of linseed diet feeding, and pigs in the 60-d treatment group had the greatest ADG and G:F. The mRNA expression of PPARgamma in loin muscle and spleen increased linearly (P < 0.01) with the duration of linseed diet feeding, whereas its expression in adipose tissue was not affected (P = 0.095). Tumor necrosis factor-alpha and IL-6 mRNA expression in muscle, adipose, and spleen, as well as serum concentration of TNF-alpha, decreased linearly (P < 0.01) with the duration of linseed diet feeding. Peroxisome proliferator-activated receptor-gamma mRNA abundance was negatively correlated with IL-1beta, IL-6, and TNF-alpha mRNA abundance both in muscle (R(2) = 0.63, P < 0.001) and in spleen (R(2) = 0.69, P < 0.001), and PPARgamma mRNA expression in spleen (R(2) = 0.59, P < 0.01) and muscle (R(2) = 0.52, P < 0.05) was negatively correlated with serum TNF-alpha concentration. There were also significant quadratic relations between ADG and expression of PPARgamma (P < 0.05) and splenic TNF-alpha (P < 0.05). These data suggest that intake of n-3 PUFA from the linseed diet led to significant decreases in the expression of proinflammatory cytokine genes, which may stimulate growth in growing-finishing barrows, at least in part, through a PPARgamma-dependent mechanism.

Transvaginal ultrasound-guided aspiration and surgical starch sclerosis therapy in recurrent pelvic inclusion cysts--report of two cases.

Pelvic inclusion cyst is one of the complications of major abdominal surgery. Treatment of the symptomatic mass is either surgical or needle aspiration. However, recurrence of the condition is still not uncommon. Repeated surgery or aspiration could result in serious morbidity. We report two cases of recurrent pelvic inclusion cyst which were successfully treated with transvaginal aspiration and sclerosing therapy with starch powder from surgical gloves and normal saline solution. The aspirations were guided by transvaginal ultrasound and on local paracervical anesthesia. Starch and normal saline solution were injected through the aspiration needle for irrigation three times. One patient with a large cyst needed two treatment courses, the other only one. Both patients had no discomfort both intra- and postoperatively. A Follow-up ultrasonography proved no recurrence to date.