[The effects of robotic-assisted versus laparoscopic-assisted radical right hemicolectomy on short-term outcome and long-term prognosis based on propensity score matching].

Objective: To compare the short-term and long-term outcomes between robotic-assisted and laparoscopic-assisted radical right hemicolectomy in patients with adenocarcinoma of the right colon. Methods: Retrospective review of a prospectively collected database identified 288 right colon cancer patients who underwent either robotic-assisted (n=57) or laparoscopic-assisted right hemicolectomy (n=231) between October 2014 and October 2020 at Department of Gastrointestinal Surgery, the Affiliated Hospital of Qingdao University. There were 161 males and 127 females, aging (60.3±12.8) years (range: 17 to 86 years). After propensity score matching as 1∶4 between robotic-assisted and laparoscopic-assisted right hemicolectomy, there were 56 cases in robotic group and 176 cases in laparoscipic group. Perioperative outcomes and overall survival were compared between the two groups using t test, Wilcoxon rank sum test, χ2 test, Fisher exact test, Kaplan-Meier method and Log-rank test, respectively. Results: The total operative time was similar between the robotic and laparoscopic group ((206.9±60.7) minutes vs. (219.9±56.3) minutes, t=-1.477, P=0.141). Intraoperative bleeding was less in the robotic group (50 (20) ml vs. 50 (50) ml, Z=-4.591, P<0.01), while the number of lymph nodes retrieved was significantly higher (36.0±10.0 vs. 29.0±10.1, t=4.491, P<0.01). Patients in robotic group experienced significantly shorter hospital stay, shorter time to first flatus, and defecation (t: -2.888, -2.946, -2.328, all P<0.05). Moreover, the overall peri-operative complication rate was similar between robotic and laparoscopic group (17.9% vs. 22.7%, χ²=0.596,P=0.465). The 3-year overall survival were 92.9% and 87.9% respectively and the 3-year disease-free survival rates were 83.1% and 82.6% with no statistical significance between the robotic and laparoscopic group (P>0.05). Conclusions: Compared to laparoscopic-assisted right hemicolectomy, robot-assisted right hemicolectomy could improve some short-term clinical outcomes. The two procedures are both achieving comparable survival.

[Situation of coal dust hazards and its damage of pulmonary function of a large-scale coal mine enterprise of Shanxi province].

Objective: To investigate the status of coal dust hazard classification and lung function damage in a large coal mine in Shanxi Province. Methods: From January to June in 2019, 51 coal dust posts and 598 workers exposed to coal dust were selected from a large coal mine enterprise in Shanxi Province. The coal dust (exhaled dust) samples were collected and tested, and the hazard classification index of coal dust (exhaled dust) was calculated. The jobs exposed to coal dust (exhaled dust) were divided into relatively harmless, mild, moderate and severe hazard posts, and the corresponding workers were divided into relatively harmless group, mild, moderate and severe hazard groups. The forced expiratory volume (FEV1) , forced vital capacity (FVC) and forced expiratory volume/forced vital capacity (FEV1/FVC) in the first second were measured. Spearman rank correlation method was used to analyze the relationship between the hazard grade of coal dust and lung function. Results: Among 51 coal dust (exhalation) posts, 13 coal dust (exhalation dust) exceeded the standard (25.5%) . 168 cases (34.78%) had abnormal pulmonary function. Compared with the relatively harmless group, the proportion of abnormal pulmonary function of workers in mild, moderate and severe hazard groups were higher, FEV1, FVC, FEV1/FVC values were lower, the differences were statistically significant (P<0.05) . The rank of coal dust (exhaled dust) was negatively correlated with FEV1, FVC and FEV1/FVC (P<0.01) . Conclusion: Attention should be paid to the supervision and management of relatively harmless and slightly harmful coal dust posts. FVC may be one of the lung function indexes sensitive to coal dust exposure.

MiR-9-5p could promote angiogenesis and radiosensitivity in cervical cancer by targeting SOCS5.

OBJECTIVE: The aim of this study was to investigate the molecular mechanism of miRNA-9-5p in cervical cancer. PATIENTS AND METHODS: The expression level of microRNA-9-5p (miR-9-5p) in cervical cancer (CC) tissues and cell lines was examined by quantitative Real Time-Polymerase Chain Reaction. Cells were transfected with Lipofectamine 3000. Cell proliferation was measured by Cell Counting Kit-8 (CCK-8). Invasion assays were performed in 24-well transwell chambers system with 8 µm pores. Cell invasion was evaluated by transwell assay. Western blot was used to detect the changes of epithelial-mesenchymal transition (EMT) and SOCS5. The effects of miR-9-5p on tubule formation were examined under different doses of γ radiation. Immunohistochemistry assay was used to analyze the protein expression of SOCS5. Fluorescence microscopy analysis was used to measure autophagosomes after cells treated with γ irradiation. RESULTS: From the Cancer Genome Atlas (TCGA) database, the expression of miR-9-5p was significantly higher in cervical cancer patients than in the negative ones, and it was verified in 22 paired of lymph node-positive patient tissues and negative. The overexpression of miR-9-5p promoted proliferation and invasion of cervical cancer cells in vitro and primary tumor growth in vivo. MiR-9-5p reduced the tubule generation after the radiation dose of 4Gy. Besides, we identified SOCS5 as the target of miR-9-5p, and the overexpression of SOCS5 could inhibit miR-9-5p mimics from promoting tubule formation. CONCLUSIONS: MiR-9-5p could promote proliferation and invasion of CC cells in vitro and in vivo. MiR-9-5p could affect angiogenesis and radiosensitivity of CC cells by targeting SOCS5.

Effect of weaning age on cortisol release in piglets.

The effect of weaning age on the adrenal cortex, which plays a vital role in the stress response, is currently unknown. Therefore, plasma adrenocorticotropic hormone (ACTH) and cortisol levels, weights and relative weights of adrenal glands, and steroidogenesis-related protein and enzyme expression levels in piglets weaned on different days were determined. Piglets weaned at 35 days had significantly lower ACTH levels than those weaned at 14 or 21 days, and cortisol levels of piglets weaned at 21, 28, and 35 days were significantly lower than those of piglets weaned on day 14. Adrenal gland weights of piglets weaned at 28 and 35 days and relative adrenal gland weights of piglets weaned at 35 days were significantly lower than those of piglets weaned at 14 days. However, no significant difference was detected in the expression of melanocortin-type 2 receptor mRNA, which is associated with weaning age. Steroidogenic acute-regulatory (StAR) mRNA and cholesterol side-chain cleavage cytochrome P450 mRNA expression levels in piglets weaned at 28 and 35 days were significantly lower than in those weaned at 14 or 21 days, and P450 11ß mRNA expression levels in piglets weaned at 28 and 35 days were significantly lower than in those weaned at 14 days. Therefore, early-weaned piglets exhibited increased adrenal gland weights and StAR and steroidogenic enzyme expression, all of which contributed to high cortisol levels. The high plasma ACTH and cortisol levels in early-weaned piglets indicate that these animals would be greatly affected by stress.

Relation between therapy options for Graves' disease and the course of Graves' ophthalmopathy: a systematic review and meta-analysis.

BACKGROUND: The relation between therapy options for Graves' disease (GD) and the course of Graves' ophthalmopathy (GO) are still controversial. Our aim was to compare the occurrence of development or worsening of GO in patients who were treated with antithyroid drugs (ATDs) or radioactive iodine (RAI) or thyroidectomy (TX). METHODS: We conducted a comprehensive search of the Embase and PubMed database. Odds ratio (OR) was used as a measure of the effect of therapy options for GD on the risk of development or worsening of GO. The analysis was further stratified by factors that could affect the treatment effects. RESULTS: Nine trials involving 1773 patients were included. RAI therapy showed a significant effect on the risk of development or worsening GO compared with ATD (OR 2.25; 95 % CI 1.61-3.14; P < 0.00001). Glucocorticoid prophylaxis was effective in preventing GO development or worsening (0.40; 0.23-0.68; P = 0.002); especially for patients with preexisting GO (0.41; 0.23-0.73; P = 0.002). At 3 months, showed GO to be improved in 17 TX and 21 total thyroid ablation (TTA) patients, with no significant difference between the two groups; between 6 and 12 months, TTA did show significant beneficial effect on the improvement of GO (6.02; 2.80-12.96; P < 0.00001); GO was found to be inactive in a significantly higher percentage of patients in the TTA (2.17; 1.04-4.52; P = 0.04). CONCLUSION: Radioiodine therapy is a significant risk factor for development or worsening of GO in GD. But GO progression can be prevented by prophylactic glucocorticoids in patients with preexisting GO. Compared with TX alone, TTA induces an earlier and steadier GO improvement in patients with mild to moderate-severe and active GO. Whether this is sufficient to offer TTA to patients needs further investigation.

First Report of Ilyonectria robusta Causing Rusty Root of Asian Ginseng in China.

Asian ginseng (Panax ginseng) is an economically important perennial herb, mainly cultivated in Jilin Province, China. In September 2013, Asian ginseng plants in Jilin showed rusty root symptoms. Typical symptoms included rusty superficial lesions of irregular shapes and margins. Ten symptomatic roots were collected from each of five fields for investigation. To isolate the pathogen, root epidermal tissues with typical lesions were excised, surface-sterilized, and placed on potato dextrose agar (PDA) amended with 50 µg/ml tetracycline. After incubation at 20 ± 1°C in the dark for a week, 18 single-spore isolates out of 50 samples were obtained and identified as Ilyonectria robusta (A.A. Hildebr.) A. Cabral & Crous based on morphological characters and DNA sequence analysis (1). After incubating 7 days on PDA in the dark at 20°C, colonies were cottony to felty in texture and orange white to brownish grey in color with average diameters of 60 ± 3 mm. Isolates were cultured on synthetic nutrient-poor agar for conidial measurements. Macroconidia formed on simple conidiophores predominately, with mostly one and occasionally up to three septa, and were cylindrical with both ends broadly rounded. Macroconidia varied in size depending on the number of cells as follows: one-septate, 7.0 ± 0.6 × 27.7 ± 2.7 µm; two-septate, 7.3 ± 0.7 × 33.3 ± 2.1 µm; three-septate, 7.4 ± 0.6 × 33.4 ± 2.2 µm. Microconidia that formed on complex conidiophores were ellipsoid to ovoid and ranged in size from aseptate 3.7 ± 0.5 × 8.7 ± 1.1 µm to one-septate 5.0 ± 0.6 × 13.1 ± 1.6 µm. Brown chlamydospores were abundantly produced on PDA, globose to subglobose in shape, and in size of 10.9 ± 1.3 × 11.8 ± 1.5 µm (n ≥ 30 observations per structure for each measurement). The isolates were further classified by amplifying and sequencing the ITS1-5.8S rRNA-ITS2 region and histone H3 gene with primer pairs ITS5 and ITS4 (4), and H3-1a and H3-1b (3), respectively. Sequences of the two loci (GenBank Accession Nos. KM015300 and KM015299) showed 100% identity among the three examined isolates and the published I. robusta isolates (JF735268 and JF735517). To confirm the pathogenicity, bare roots of 3-year-old Asian ginseng were inoculated with mycelial plugs of three isolates of I. robusta selected randomly. Four roots were inoculated as replicates for each isolate with pathogen-free agar plugs as a control. One week post-inoculation in the dark at 20 ± 1°C, all the inoculated ginseng roots showed light-brown to dark-brown lesions. I. robusta was recovered from symptomatic roots and confirmed by analyzing the DNA sequence of the histone H3 gene. The inoculation experiment was repeated, and both trials showed the same results. The ginseng tissue under the control agar plugs remained symptomless, and no fungi were isolated. To our knowledge, this is the first report of I. robusta causing rusty root of P. ginseng in China (1,2,5). References: (1) A. Cabral et al. Mycol. Prog. 11:655, 2012. (2) I. Erper et al. Eur. J. Plant Pathol. 136:291, 2013. (3) N. L. Glass et al. Appl. Environ. Microbiol. 61:1323, 1995. (4) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990. (5) X. Lu et al. Plant Dis. 98:1580, 2014.

Lactoferrin mRNA expression in mouse mammary glands during pregnancy and lactation.

Lactoferrin (Lf) is an iron-binding glycoprotein that is produced by mucosal epithelial cells in mammals. Lf has non-immune natural defense functions and biological functions in addition to and distinct from its role in regulating inflammatory responses. Lf also improved some physiological and immunological parameters. Lf is a biomarker for monitoring medical treatment in inflammatory bowel diseases. Current LF research focuses on iron absorption, antimicrobial activity, and the modulation of iron metabolism during inflammation. No systematic research about Lf expression levels in mouse mammary glands during pregnancy and lactation exists. We investigated Lf mRNA expression levels in mouse mammary glands by collecting samples on days 1, 6, 12, and 18 of pregnancy and lactation (six mice per group). The expression levels of Lf mRNA were measured by semi-quantitative reverse transcription polymerase chain reaction using GAPDH as an internal control. Lf mRNA was not expressed in mammary glands on days 1, 6, and 12 of pregnancy, but it was expressed on day 18 (IOD: integrated optical density; Lf(IOD)/GAPDH(IOD) = 0.46). Lf expression levels were higher during lactation stages than during pregnancy stages, and it stabilized at 0.71-0.73 (Lf(IOD)/GAPDH(IOD)) from day 1 to 12 of lactation; however, the difference was not significant (P > 0.05). At day 18 of lactation, Lf expression began to decline (Lf(IOD)/GAPDH(IOD) = 0.61), but the difference was not significant (P > 0.05). Based on these results, the variation in Lf expression levels during developmental stages may be related to its regulatory role in mouse mammary gland immunity.

Phospholipases A-II (PLA2-II) induces acute pancreatitis through activation of the transcription factor NF-kappaB.

OBJECTIVES: Acute pancreatitis (AP) is an inflammatory disease of the pancreas characterized by local inflammation. Secretory phospholipases A-II (sPLA2-II) have been implicated in triggering AP, but their exact role to evoke AP is largely unknown. NF-kB activation has previously been shown to induce acute pancreatitis. The aim of this study is to explore that PLA2-II triggers AP by activation of NF-kB and the expression of inducible inflammatory mediators. MATERIALS AND METHODS: Acute pancreatitis in vivo was induced in Wistar rats by retrograde infusion of 4% sodium taurocholate (TAC) into the pancreatic duct. Then the Wistar rats were devided into 2 groups: (1) PLA2 II-specific siRNA was subsequently administrated subcapsularly after infusion of TAC. (2) One hour before the intraductal injection of TCA, the rats were treated with PDTC 100 mg/kg twice i.p. in 1 h interval. Induction of pancreatitis was confirmed by histopathology, NF-kB activity and expression in pancreas was detected by EMSA and immunohistochemistry. Inflammatory mediators such as the tumor necrosis factor-alpha (TNF-α), interleukin (IL)-1beta, intercellular adhesion molecule-1 (ICAM-1), IL-6 and IL-8 in blood was detected by ELISA. The severity of the disease and the mortality were observed. RESULTS: We demonstrated that TAC specifically induces pancreatitis, induces PLA2-II expression and activates NF-kappaB and proinflammatory cytokine synthesis in the pancreas of rats. sPLA2-II siRNA transfection blocked NF-kappaB activation and proinflammatory cytokine expression and relieved pancreatitis severity. PDTC treatment blocked NF-kappaB activation and proinflammatory cytokine expression. Pretreatment with PDTC or PLA2 II-specific siRNA transfection improved the survival of the rats. CONCLUSIONS: These findings suggest that PLA2-II induces acute pancreatitis through activation of the transcription factor NF-kappaB. siRNA mediated gene knockdown of PLA2-II relieves pancreatitis severity at least partly mediated by the inhibition of NF-kappaB activation and proinflammatory cytokine synthesis.

Clinical significance of serum matrix metalloproteinase-13 levels in patients with esophageal squamous cell carcinoma (ESCC).

OBJECTIVES: Matrix metalloproteinase-13 (MMP13) is a highly regulated zinc-dependent endopeptidase and has been reported to be associated with vascular invasion and lymph node metastasis and predicts poor outcome for relatively early stage in esophageal squamous cell carcinoma (ESCC) patients. However, the role of the serum MMP-13 levels in ESCC is still unknown. In the present study, we investigate the clinical significance of MMP-13 levels in patients with ESCC. PATIENTS AND METHODS: The serum level of MMP-13 was measured with commercially available ELISA kit in 112 healthy controls and 141 ESCC patients prior to surgical resection. Statistical associations between clinicopathological observations and MMP-13 levels were determined using the Mann-Whitney U test. The clinical value of MMP-13 level as a prognostic parameter was evaluated using the Cox's proportional hazards model. RESULTS: The results showed compared with the healthy control group (74.5±12.3 ng/ml), ESCC patients tended to have significantly higher serum MMP-13 concentrations (86.2 ± 14.6 ng/ml) (p < 0.05). Elevation of MMP-13 levels (≥ 76.4 ng/ml) was observed in 61.7% (87/141) of patients with ESCC, and 18.4% (26/141) in healthy controls. MMP-13 levels were associated with lymph node metastasis (p < 0.001), distant metastasis (p < 0.001), histological differentiation (p = 0.026), T classification (p < 0.005), but not with the tumor size, clinical stage, age and gender of the patients or tumor location. Multivariate analysis revealed that patients with an elevated level of MMP-13 (≥ 76.4 ng/ml) had significantly lower 5 year survival rate than those with non-elevated MMP-13 (< 76.4 ng/ml, log-rank p < 0.001). CONCLUSIONS: It is suggested that the elevated level of preoperative MMP-13 was found to associate with tumor progression and poor survival in patients with ESCC.

First Report of Rhexocercosporidium panacis Causing Rusty Root of Panax ginseng in Northeastern China.

In northeastern China, Asian ginseng (Panax ginseng) roots exhibited reddish brown lesions of various sizes, irregular shapes, and diffuse margins, typical of rusty root disease. The lesions remain superficial, smooth, and limited to the epidermal and peridermal tissues. In September 2013, 10 symptomatic roots were collected from each of three fields in Jilin and Heilongjiang provinces. One piece of symptomatic skin tissue from each root was excised, surface-disinfested in 1% NaClO for 3 min, rinsed three times with sterile water, and then placed on tetracycline-amended (50 µg/ml) potato dextrose agar. After incubation at 22 ± 1°C in the dark for a week, small olivaceous black colonies developed from the symptomatic tissue from five of the 30 samples. No spores were observed. A single hyphal tip of each colony was transferred to a fresh V8 agar plate to purify the culture. Two-week-old colonies on V8 agar were olivaceous gray, and 42 to 46 mm in diameter with an outer white margin (3 to 5 mm wide). Conidia produced in V8 broth after 3 weeks with a 12-h photoperiod were straight and hyaline, cylindrical or subcylindrical with no or one septum. Mature conidia were 12.8 to 21.8 × 2.2 to 4.5 µm (mean 18.2 × 3.0 µm, n = 100 conidia for each of three isolates). Three isolates selected randomly were further identified by analyzing the partial sequences of the ITS region of rDNA with primers ITS4 and ITS5 (5), and partial sequences of ß-tubulin with the primers tub2F and tub2R (1). Sequences of the three isolates (GenBank Accession Nos. KJ149287, KJ149288, and KJ149290 to 93) showed 99% to 100% homology with previously identified and deposited Rhexocercosporidium panacis isolates (DQ2499992 and DQ457119) for both loci (3). Therefore, the three isolates were identified as R. panacis and deposited in China General Microbiological Culture Collection Center (CGMCC3.17259 to 61). Pathogenicity of R. panacis in Asian ginseng was investigated using these three isolates as described previously with slight modifications (4). Bare roots of 3-year-old Asian ginseng were surface-disinfested as described above, and inoculated with mycelial plugs (4 mm diameter) cut from the margin of actively growing colonies of the isolates on V8 agar. Three mycelial plugs were placed on each root at 3-cm intervals and four roots (replicates) were inoculated for each isolate. Four additional roots were inoculated with non-colonized agar plugs as control. The treated roots were placed on moist filter paper in an enamel tray. The plates were sealed with plastic wrap to prevent desiccation and incubated in the dark at 18 ± 1°C. Four weeks post inoculation, all the inoculated ginseng roots showed red-brown lesions, which turned to dark red or black over time. R. panacis was recovered from symptomatic roots for all isolates and confirmed by ITS sequence analysis. The mock-inoculated control roots remained symptomless and no R. panacis was isolated. The inoculation experiment was repeated and showed the same results. R. panacis was reported in 2006 to infect roots of Panax quinquefolius (2,3,4). To our knowledge, this is the first report of R. panacis causing rusty root of P. ginseng. References: (1) P. R. Hirsch et al. Mycol. Res. 104:435, 2000. (2) Z. K. Punja et al. Can. J. Plant Pathol. 35:503, 2013. (3) R. D. Reeleder. Mycologia. 99:91, 2007. (4) R. D. Reeleder et al. Phytopathology 96:1243, 2006. (5) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

S100A4 regulates migration and invasion in hepatocellular carcinoma HepG2 cells via NF-κB-dependent MMP-9 signal.

BACKGROUND AND OBJECTIVES: We previously showed that the calcium-binding protein S100A4 is overexpressed and related to metastasis in hepatocellular carcinoma (HCC). However, whether S100A4 participates in the regulation of metastasis and its mechanisms in HCC is mostly unknown. Given the associations of S100A4, nuclear factor-kB (NF-kB/RelA) and MMP-9 with metastasis in a variety of malignancies, we explored a potential role of S100A4 in HCC metastasis and its mechanism. METHODS: 20 patients with HCC invasion (Lymph node metastasis, microvascular invasion, major portal vein invasion and intrahepatic metastasis) and 20 patients without HCC invasion were included. These tissues were detected for the expression of S100A4, NF-kB/RelA and MMP-9 by immunohistochemistry and quantitative real time polymerase chain reaction (Q-PCR). Correlation between the expressions of S100A4, NF-kB/RelA and MMP-9 with the invasion was analysed. The expressions of S100A4, nuclear factor-kB and MMP-9 was evaluated in HepG2 cells by western blot and immunohistochemistry. HepG2 cells were stably transfected with S100A4-specific small interfering RNA (S100A4 siRNA) to knockdown of S100A4, then transiently transfected with S100A4 cDNA to rescure the S100A4 level and evaluated for effects on invasion and expression analysis for molecules involved in invasion. After the HepG2 cells recurred the S100A4 levels, the HepG2 cells was treated with 5 µM Pyrrolidine Dithiocarbamate (PDTC) (a selective NF κ B inhibitor) to inhibit the NF-kB activity, or treated with Batimast (BB94: a MMPs inhibitor) to inhibit the MMP-9 activity. The expression analysis for molecules involved in invasion was analyzed. RESULTS: A significant increase of S100A4, NF-kB/RelA and MMP-9 expression in HCC tissues with invasion than that of without invasion. A positive correlation was observed between S100A4, NF-kB/RelA, MMP-9 and invasion, respectively. In addition, S100A4 was positively correlated with NF-kB and MMP-9. S100A4 siRNA mediated knockdown of S100A4 in HepG2 cells resulted in significant reduction in the NF-kB activity and MMP-9 expression, and dramatically decreased its invasion. Moreover, the HepG2 cell metastatic potential was rescued by overexpression of S100A4 completely, at the same time, the NF-kB activity and MMP-9 expression was also increased. Pretreatment with PDTC or BB94 was observed to significantly reduce NF-kB activity and MMP-9 expression and dramatically decreased S100A4 -induced invasion. CONCLUSIONS: Our findings indicate that S100A4 contributes to HCC metastasis by activation of NF-kB dependent MMP-9 expression, suggesting S100A4 as a novel diagnostic biomarker and therapeutic target in HCC.

Knockdown of PLA2G2A sensitizes gastric cancer cells to 5-FU in vitro.

BACKGROUND-AIM: Elevated expression of the PLA2G2A phospholipase in gastric cancer (GC) is associated with improved patient survival. PLA2G2A is also an important regulator of proliferation, invasion and metastasis in GC. However, no relation about PLA2G2A and chemosensitivity in GC cells was reported. 5-Fluorouracil (5-FU) is widely used for treatment of advanced gastric cancer. However, it is common for such patients to develop resistance to 5-FU, and this drug resistance becomes a critical problem for chemotherapy. The mechanisms underlying this resistance are largely unknown. In the present study, we investigated whether PLA2G2A could confer 5-FU resistance or sensitise in GC cells in vitro. MATERIALS AND METHODS: The 5-FU sensitivity of GC cell lines SGC-7901, MKN-45, RF-48, N87, AGS, MKN-28, RF-1, MGC-803 were determined by MTT assays. PLA2G2A levels were determined by western blot assays. The effects of 5-FU on PLA2G2A expression were determined in vitro. PLA2G2A was inhibited by silencing of the PLA2G2A using small interfering RNA in vitro. PLA2G2A was overexpressed by transfection of full-long PLA2G2A cDNA in vitro, and the effects were evaluated on 5-FU sensitivity. RESULTS: The cell lines SGC-7901, MKN-45, RF-48 and N87 were sensitive, whereas AGS, MKN-28, RF-1 and MGC-803 were resistant to 5-FU. Significant correlation was observed between basal PLA2G2A and 5-FU sensitivity. Silencing of PLA2G2A increased 5-FU killing in 5-FU-treated cells, and overexpression of PLA2G2A decreased 5-FU killing in 5-FU-treated cells. CONCLUSIONS: PLA2G2A was correlated with sensitivity to 5-FU. Silencing of PLA2G2A was sensitive to 5-FU treatment. Thus, PLA2G2A may be a useful therapeutic target for a subset of gastric cancers.

Downregulation of CD24 inhibits invasive growth, facilitates apoptosis and enhances chemosensitivity in gastric cancer AGS cells.

BACKGROUND AND AIM: The human CD24 antigen is a small heavily glycosylated cell surface protein, which is expressed in a large variety of solid tumors,including gastric cancer. Enriched on the surface of many tumor cells,CD24 promotes tumor growth,invasion and metastasis and confers resistance to some chemotherapeutic drugs.In this study, we investigated the possible effect of CD24 suppression on proliferation, apoptosis, migration, invasion and chemosensitivity of gastric cancer (GC) cells. MATERIALS AND METHODS: We down-regulated CD24 expression by RNA interference and investigated the effects on proliferation, apoptosis, chemosensitivity to doxorubicin, malignant and metastatic potential of a human gastric cancer cell line, AGS, CD24-suppressed clones, AGS-CD24-siRNA-C2, AGS-CD24-siRNA-C4 and AGS-CD24-siRNA-C5 in vitro. We evaluated the effects of CD24 suppression in vivo on xenograft tumor growth and metastatic properties following tail iv AGS-CD24-siRNA-C2, AGS-CD24-siRNA-C4 and AGS-CD24-siRNA-C5 clones. We also investigated the effect of CD24-siRNA followed by doxorubicin administration treatment on the xenograft tumor growth. RESULTS: CD24 suppressed showed significantly decreased proliferation, invasion and increased apoptosis as well as increased chemosensitivity to doxorubicin in vitro and in vivo. CONCLUSIONS: CD24 involves in proliferation, invasion and chemosensitivity of human gastric cancer cell line AGS, and that down-regulation of CD24 protein expression decreases the metastatic potential and increases chemosensitivity of gastric cancer (GC) cells. Thus, CD24 may be a promising therapeutic target for gastric cancer.

Effect of phospholipase A2 silencing on acute experimental pancreatitis.

BACKGROUND AND OBJECTIVES: The group II phospholipase A2 (PLA2 II) in blood has been reported to increase in acute pancreatitis and to reflect the severity of pancreatitis. Here we investigated the effect of inhibition of PLA2 using siRNA gene knockdown in vitro and in an in vivo model of experimental pancreatitis. MATERIALS AND METHODS: Pancreatic acinar cell line AR42J in vitro was cultured with lysophosphatidylcholine (lyso-PC) (50 uM) to induce expression of PLA2 with subsequent transfection of siRNA into stimulated AR42J cells. Acute pancreatitis in vivo was induced in Sprague Dawley rats by retrograde infusion of 4% sodium taurocholate (NaT) into the pancreatic duct. PLA2 II -specific siRNA was subsequently administrated, subcapsularly, after infusion of NaT. Controls included administration of scrambled siRNA (SC-RNA) or vehicle alone. After 24hr, pancreata were harvested and assessed for worsening pancreatitis by histopathology; The serum levels of PLA2 II and inflammatory mediators were analyzed. In both models endogenous PLA2 II gene expression was assessed at 24 hrs using real time RT-PCR. RESULTS: In vitro, PLA2 II protein and mRNA levels were reduced in cells treated with PLA2-II siRNA when compared with control treatment. In vivo, induction of pancreatitis was confirmed by histopathology, inflammatory mediators such as the tumor necrosis factor-alpha, interleukin (IL)-1beta, IL-6 and IL-8. PLA2 expression were reduced 69% in siRNA treated rats, compared with controls. Serum inflammatory mediators levels decreased after administration of siRNA compared with vehicle control (p < 0.05,respectively). CONCLUSIONS: siRNA mediated gene knockdown of PLA2-II appeared to relieve pancreatitis severity. PLA2-II may serve as a novel and effective therapeutic target for acute pancreatitis.

Prognostic significance of PUMA in pancreatic ductal adenocarcinoma.

OBJECTIVES: To investigate retrospectively whether alterations of p53 upregulated mediator of apoptosis (PUMA) protein levels and somatic mutations of the PUMA gene are characteristic of pancreatic ductal adenocarcinoma (PDAC). METHODS: Immunohistochemical analyses of PUMA were performed in pancreatic tumour tissue samples, and paired normal pancreatic tissue samples, from patients with PDAC. Apoptosis was detected using terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labelling assay. RESULTS: A total of 70 patients with PDAC had samples available; 49 cases (70.0%) had high PUMA protein levels. PUMA was not detected in paired normal tissue samples. Significantly higher levels of PUMA protein were detected in low-grade tumours (tumour -node-metastasis stages I and II), compared with higher grade (stage III) tumours. Of the PDAC cases, the mean apoptosis index value for PUMA-positive specimens was significantly higher than that for PUMA-negative specimens. Overall survival was significantly associated with PUMA immunoreactivity. CONCLUSIONS: High levels of PUMA in PDAC tumour cells suggest that PUMA expression may play a role in pancreatic tumourigenesis.

[Methodological study on the determination of oxiracetam concentration in serum and urine by HPLC].

A method was developed for the determination of oxiracetam concentration in serum and urine by HPLC. Acyclovir was used as an internal standard. The analytical column was a stainless-steel column (30 mm x 4.6 mm ID) filled with 10 microns Bondapak NH2. A mixture of acetonitrile and water (80: 20) as mobile phase was used at a flow rate of 1 ml.min-1. Detection was performed at 210 nm. The retention times were 6.3 min for oxiracetam and 8.1 min for the internal standard. The lower detection limits were 1 microgram.ml-1 for serum and 20 micrograms.ml-1 for urine. The precision and accuracy within-day and day-to-day for both serum and urine samples ranged from 5.0 to 10.7%. The mean recoveries were 99.7 +/- 5.9% and 99.0 +/- 5.6% for human serum and urine, respectively. The results showed that the method is simple, rapid, sensitive, reliable and good enough to be used in studying the clinical pharmacokinetics of oxiracetam.