Snail Research Articles

Endemic situation of schistosomiasis in national surveillance sites of Hubei Province from 2015 to 2019

To analyze the changing tendency in the endemic situation of schistosomiasis in the national surveillance sites of Hubei Province, so as to provide the scientific evidence for facilitating the progress towards the elimination of schistosomiasis in the province. According to the National Schistosomiasis Surveillance Scheme (2014 version), a total of 65 national schistosomiasis surveillance sites were assigned in Hubei Province, and the Schistosoma japonicum infections in humans, domestic animals, snails and wild feces were monitored in these national surveillance sites from 2015 to 2019. All data pertaining to the surveillance results were descriptively analyzed. There were 151 159 person-time local residents and 70 928 person-time mobile populations screened for S. japonicum infections in the 65 national surveillance sites of Hubei Province from 2015 to 2019, and the seroprevalance of S. japonicum infections reduced from 3.45% (1 077/31 197) in 2015 to 1.57% (431/27 371) in 2019 among local residents and from 1.06% (98/9 249) in 2015 to 0.81% (116/14 318) in 2019 among mobile populations. During the period from 2015 through 2019, there were 22 egg-positive local residents and 2 egg-positive mobile populations identified in 2015, with 0.07% and 0.02% prevalence rates of S. japonicum infections, respectively. During the 5-year period, a total of 7 025 herd-time domestic animals examined for S. japonicum infections, with no infections detected, and a total of 2 035 wild feces were tested, with no egg-positives identified in the 65 national surveillance sites of Hubei Province. Snail survey was conducted covering 28 767.35 hm2 during the 5-year period, and the area of snail habitats ranged from 1 609.82 to 2 119.81 hm2. The mean density of living snails and occurrence of frames with snails reduced from 0.360 5 snails/0.1 m2 and 11.26% in 2015 to 0.175 9 snails/0.1 m2 and 8.43% in 2019, respectively, and no S. japonicum infection was found in snails during the 5-year period. In addition, no snails were found in the potential schistosomiasis transmission foci in the two national surveillance sites in the Three Gorges Dam. The overall endemic situation of schistosomiasis appears a tendency towards a decline in Hubei Province during the period from 2015 through 2019; however, the schistosomiasis transmission risk remains. Improving the surveillance system of schistosomiasis and increasing the sensitivity and performance of the surveillance system are required to provide insights into the implementation of precision control strategy and interventions in Hubei Province.

Endemic situation of schistosomiasis in national surveillance sites of Zhenjiang City from 2015 to 2019

To analyze the endemic situation of schistosomiasis in national surveillance sites of Zhenjiang City from 2015 to 2019, so as to provide the scientific basis for the consolidation of schistosomiasis control achievements and timely treatment of risk factors. A total of 7 national schistosomiasis surveillance sites were assigned in Zhenjiang City from 2015 to 2019. According to the National Scheme for Schistosomiasis Surveillance (2014 version), Schistosoma japonicum infections were monitored in local residents, floating populations and livestock, and snail habitats were surveyed in Zhenjiang City from 2015 to 2019. The results of schistosomaisis surveillance in Zhenjiang City from 2015 to 2019 were analyzed. There were 15 848 and 7 779 person-times of local residents and floating populations receiving serological screening of schistosomiasis in 7 national surveillance sites of Zhenjiang City from 2015 to 2019, and the seroprevalence rates were 0.95% to 1.75% and 0.14% to 0.91% among local residents and floating populations, respectively; however, no egg-positives were identified. A total of 1 885 herd-times of livestock were detected, and no egg-positives were found. During the 5-year period, snail survey was conducted in snail habitats and suspected snail habitats covering 3 333.79 hm2, and there were 108.71 hm2 snail habitats identified, including 50.97 hm2 re-emerging snail habitats. The density of living snails was 0.007 to 0.125 snails/0.1 m2 from 2015 to 2019, which showed a trend for a rise year by year (χ2 = 2 732.626, P < 0.01); however, no S. japonicum infections were detected in snails. The endemic situation of schistosomiasis has been maintained at a low level in Zhenjiang City, and schistosomiasis control achievements are consolidated. However, snails are widely distributed in the city, and there are still risk factors associated with schistosomiasis transmission. Further integrated interventions require to be reinforced to timely manage the risk of transmission of schistosomiasis and promote the progress towards elimination of schistosomiasis.

Endemic situation of schistosomiasis in national surveillance sites of Hunan Province from 2015 to 2019

To analyze the changes in the endemic situation of schistosomiasis in national surveillance sites of Hunan Province, so as to provide scientific basis for the development of the schistosomiasis elimination programme in the province. According to the requirements of the National Guidelines for Schistosomiasis Surveillance in China (2014 version), a total of 41 national schistosomiasis surveillance sites were assigned in all disease-endemic counties (districts) across Hunan Province. During the period between 2015 and 2019, Schistosoma japonicum infections were monitored in local residents, mobile populations and livestock, and snail status was monitored. The morbidity due to schistosomiasis and snail status was compared between years. The sero-prevalence of S. japonicum infections was 2.57% and 1.56% in local residents and mobile populations in national surveillance sites of Hunan Province from 2015 to 2019, respectively, and the sero-prevalence appeared a tendency towards a decline over years. A higher sero-prevalence rate of S. japonicum infections was seen in men than in women (P < 0.01). During the 5-year study period, the sero-prevalence rate of human S. japonicum infections appeared a tendency towards a decline in the marshland, embankment, inner embankment and hilly types of endemic areas over years. There were 44 and 19 egg -positives detected in local residents and 5 and 1 egg-positives in mobile populations in 2015 and 2016 respectively. A total of 9 346 domestic animals were monitored from 2015 to 2019, and 6 egg-positives were detected in 2015 and 2016 (all were bovine). A total of 0.155 billion m2 settings were surveyed from 2015 to 2019, and the mean density of living snails appeared a tendency towards a decline over years, with a 45.79% reduction in 2019 as compared to 2015. However, no S. japonicum infections were identified in snails during the 5-year period. A total of 1 469 mixed snail samples were detected using loop-mediated isothermal amplification (LAMP), and 6 positive snail samples were identified in 2015 (one sample) and 2017 (5 samples). The overall endemic situation of schistosomiasis appears a tendency towards a decline in Hunan Province, and the prevalence of S. japonicum infections is at a low level in humans and livestock; however, there is still a risk of schistosomiasis transmission. Improvements of health education, intensification of schistosomiasis examinations in mobile populations and reinforcement of the surveillance-response system is required to consolidate the achievements of schistosomiasis control in Hunan Province.

Endemic situation of schistosomiasis in national surveillance sites of Anhui Province from 2015 to 2018

To analyze the endemic situation of schistosomiasis in national surveillance sites of Anhui Province from 2015 to 2018, so as to provide scientific evidence for schistosomiasis control and prevention in Anhui Province. According to the National Schistosomiasis Surveillance Programme (2014 version), a total of 51 national schistosomiasis surveillance sites were assigned in Anhui Province in 2015, and Schistosoma japonicum infections in humans and livestock and snail distribution were monitored from 2015 to 2018. A total of 89 638 local residents and 42 609 mobile populations received serological screening of schistosomiasis in 51 national surveillance sites of Anhui Province from 2015 to 2018, and the sero-prevalence of S. japonicum infections was 1.41% to 3.69% in local residents and 0.84% to 2.13% in mobile populations, respectively. There were 5 egg-positive local residents and 1 egg-positive mobile populations detected in 2015, with occupations of farmers and fishermen. There were 6 405 livestock detected for S. japonicum infections, and no egg-positives were identified. Among an area of 12 661 hm2 surveyed in the national schistosomiasis surveillance sites of Anhui Province from 2015 to 2018, the areas of snail habitats were 2 461.27 to 2 628.96 hm2, andthemeandensityoflivingsnailswas 0.3757 to 0.4330 snails/0.1 m2, with no S. japonicum infections identified in snails. The endemic situation of schistosomiasis is at a low level in Anhui Province; however, the risk of schistosomiasis transmission remains in local regions of the province. The construction of the surveillance-responsesystemshouldbereinforcedtoconsolidatetheachievementsofschistosomiasis control in Anhui Province.

Involvement of epithelial-mesenchymal transition-inducing transcription factors in the mechanism of Helicobacter pylori-induced fibroblasts activation.

Gastric cancer (GC) originating from the lining of the stomach is one of the most frequent malignancies in humans. The most efficient method giving hope of full recovery from GC is gastric resection combined with adjuvant chemotherapy and radiotherapy. However, over 50% of patients after treatment suffer from recurrence and peritoneal metastasis. The bacteria Helicobacter pylori (Hp) is nowadays considered as the major pathogen capable of colonizing gastric mucosa. This bug causes deregulation of multiple signaling pathways including the activation of nuclear factor kappaB (NFκB) and Janus kinase/signal transducers and activators of transcription (JAK/STAT) responsible for inflammation and development of carcinogenic cascade. The pathomechanism of these changes remains little understood, but the Hp virulence factors affecting mainly gastric epithelium have been postulated. Nevertheless, the changes associated with inflammation progressing to cancer are not only limited to epithelial cells. The cells surrounding the tumor, mainly activated fibroblasts (CAFs - cancer-associated fibroblasts) create molecular microenvironment promoting tumorigenesis and cancer invasion. The downstream targets of STAT3, epithelial-mesenchymal transition-inducing transcription factors (EMT-TFs) are expressed in activated fibroblasts providing them with additional properties. Thus, our aim was to determine if the Hp strain expressing CagA and VacA cytotoxins may result in the activation/differentiation of rat gastric fibroblasts resulting in NFκB and STAT3 signaling, which could lead to EMT-TFs expression and secretome responsible for inflammatory and EMT inducing microenvironment. In this study, gastric samples were harvested from 8-week-old Spraque-Dowley rats and the primary and secondary fibroblast cultures were established. The 70% confluent secondary fibroblast cultures were infected with 1 x 109 of live Hp expressing cytotoxins CagA VacA per dish and incubated in humidified atmosphere for 3, 24, 48 and 72 hours, before the conditioned media or the cells were used for endpoint experiments. As the control, fibroblast culture in DMEM with 10% FBS and antibiotics, free from Hp infection was used. The expression of mRNA for 18S (control), toll-like receptors: TLR2 and TLR4, STAT3, NFκB p65/Rel A, inhibitor of NF-κB (Iκβ), Snail and Twist was determined by RT-PCR. The protein expression of Snail and Twist was assessed by Western blot technique. The fibroblast supernatant was collected at 72 hours from non-infected and Hp (cagA+ vacA+)-infected culture and the concentrations of interleukin 8 (IL-8), hepatocyte growth factor (HGF) and stromal derived factor-1 (SDF-1) were measured by ELISA. In fibroblasts infected with Hp (cagA+ vacA+), the significant increase of mRNA expression for both, TLR2 and TLR4, as well as STAT3, NFκB/RelA subunit was observed already after 3 hours of fibroblasts infection with Hp strain compared with control non-infected fibroblasts. Simultaneously, the significant decrease of Iκβ mRNA has been noticed starting from 48 hours after the Hp infection of fibroblasts was carried out. The strong increase in the expression of Snail1 and Twist mRNA was recorded already at 3 hours in Hp-infected fibroblasts comparing to control non-infected fibroblasts and this increase persisted at 24 and 48 hours being the most pronounced at 72 hours post incubation with Hp. The expression of Snail1 protein was observed after 3 hours post Hp infection and this increase persisted throughout entire time periods upon Hp infection. In contrast, no detectable level of Twist protein expression was observed up to 72 hours post-infection neither in control conditions nor in fibroblasts co-infected with Hp. These changes in fibroblasts were accompanied by a significant increase in the release of HGF, SDF-1 and IL-8 determined in cell supernatants collected from Hp-infected fibroblasts. These data indicate that the activation/differentiation of rat gastric fibroblasts can occur directly by Hp releasing CagA and indirectly through TLR2 and TLR4 and these effects can be mediated by transcription factors NFκB and STAT3 signaling leading to rapid Snail1 protein expression. We conclude that NFκB and STAT3 signaling together with Snail1 protein expression may activate the secretome responsible for fibroblasts inflammatory and EMT-inducing microenvironment likely serving as prerequisite for GC development.

Rhizoma Pleionis inhibits of breast cancer epithelial-mesenchymal transition by VEGF-A related angiogenesis

Objective To investigate the Rhizoma Pleionis mediating vascular endothelial growth factor A (VEGF-A)-related angiogenesis in breast cancer cells to inhibit epithelial-mesenchymal transition (EMT) in breast cancer. Methods Firstly, the proliferation activity of MDA-MB-231 and 4T1 of breast cancer cells was detected by methyl thiazolyl tetrazolium (MTT) method at different concentrations (0.2 g/ml, 0.4 g/ml, 0.6 g/ml, 0.8 g/ml, 1.0 g/ml); secondly, the specific concentration of Rhizoma Pleionis was selected and tested by Transwell test for its effect on breast cancer cell metastasis and invasion. The formation of human umbilical vein endothelial cells (HUVEC) capillary wall was measured to detect the effect of Rhizoma Pleionis on cell angiogenesis. The EMT-related protein and angiogenesis-related molecules were detected by Western blot. Results MTT results showed that Rhizoma Pleionis of 0.8 g/ml and 1.0 g/ml can inhibit the MDA-MB-231 and 4T1 cells proliferation in a dose-dependent manner, with statistically significant difference (P<0.05). Rhizoma Pleionis can inhibit the invasion of breast cancer cells and inhibit the formation of blood vessel walls in HUVECs. Western blot confirmed that the expression level of EMT-related proteins: E-cadherin was significantly increased and the expression level of N-cadherin, Vimentin and Snail was significantly decreased after treatment with Rhizoma Pleionis (P<0.05); and the level of the related molecule hypoxia inducible factor-1α (HIF-1α), VEGF-A and vascular endothelial growth factor receptor-2 (VEGFR-2) was significantly reduced (P<0.05). Conclusions Rhizoma Pleionis inhibits the epithelial-mesenchymal transition of breast cancer by influencing the angiogenesis of VEGF-A, and inhibits the invasion and metastasis of breast cancer. Key words: CREMASTRA VARIABILIS; Vascular endothelial growth factor A; Breast neoplasms; Tumor cells, cultured; Epithelial-mesenchymal transition

Role of interleukin-10 in macrophage promoted progress in pancreatic cancer

Objective To examine the role of interleukin-10 (IL-10) in macrophage induced progression of pancreatic cancer. Methods Based on the different intervention methods for IL-10 in macrophages, nude mices were divided into five groups: IL-10 overexpression group; Recombinant IL-10 group; IL-10 neutralizing antibody group; IL-10 low expression group and control group. The tumor growth and the size of metastases were measured. Epithelial-mesenchymal transition markers including proliferation cell nuclear antigen (Ki-67), E-cadherin, Vinmentin and Snail protein expressions were detected. Multiple groups were compared using one-way ANOVA. Results The volumes in overexpression and recombinant factor group were significantly larger than that in control group, which were (1 089.8±60.3), (987.4±50.2), (660.5±40.1) mm3 respectively. The difference is significant (F=20.341, P 0.05), while the neutralizing antibodies in the low expression group and the control group were significantly lower (F=11.103, P<0.05). These results were same to liver in peritoneum and diaphragm metastases. E-cadherin expression in overexpression group and recombinant factor group was significantly lower than that in control group, which were 0.070±0.003, 0.060±0.002 and 0.300±0.020, respectively (F=15.320, P<0.05). The expression of Ki-67, Vinmentin and Snail showed opposite changes: increased in the overexpression group and recombinant factor group, while decreased significantly in the neutralizing antibody and low expression group. Conclusion IL-10 plays an important role in the macrophage induced progression of pancreatic cancer. Key words: Pancreatic cancer; Macrophage; Epithelial-mesenchymal transition; Interleukin-10

Role of SATB1 in invasion and metastasis of human laryngeal squamous cell carcinoma cell line Hep-2 and its mechanisms

Objective To investigate the role of special AT-rich sequence-binding protein 1 (SATB1) in invasion and metastasis of human laryngeal squamous cell carcinoma cell line Hep-2 and its mechanism. Methods Hep-2 cells were cultured in vitro. The cell lines were divided into two groups. The cells in the experimental group was transfected with Lipofectamine 2000 liposome for SATB1 small interfering RNA (SATB1-siRNA), and the cells in the control group was transfected with Lipofectamine 2000 liposome for NC-siRNA. The before and after transfection SATB1, E-cadherin and Snail genes relative expression levels between the two groups were detected and compared using RT-PCR. Transwell cell invasion experiments were applied to evaluate the cells invasive ability of the two groups. The scratch experiments were applied to evaluate the 48 h cell migration ability of the two groups. Results The differences in the relative expression level of SATB1, E-cadherin and Snail genes in pre-transfected laryngeal squamous cell line Hep-2 between the two groups were not statistically significant (all P>0.05). Compared with the control group, the relative expression levels of the mRNA of SATB1 and Snail genes decreased in the experimental group after transfection, while the relative expression levels of the mRNA of E-cadherin gene increased, and the differences were statistically significant (all P<0.05). After the transfection, the number of transwell membrane cells and cell migration distances of the two groups of cells decreased. The values of cell number and migration distance in the experimental group were lower than those in the control group, and the differences were statistically significant (all P<0.05). Conclusions SATB1 can promote the invasion and metastasis of Hep-2 cells, and its mechanism may be related to the down-regulation of E-cadherin and the up-regulation of Snail gene expression to induce epithelial-mesenchymal transition. Key words: Laryngeal neoplasms; Carcinoma, squamous cell; Neoplasm invasiveness; Neoplasm metastasis; Special AT-rich sequence-binding protein 1

Tumor necrosis factor receptor associated factor 4 promotes invasion and metastasis of colorectal cancer cell line LoVo

Objective To study the effect of low expression of tumor necrosis factor receptor associated factor 4 (TRAF4) on invasion and migration of colorectal cancer cells. Methods Immunohistochemistry was used to detect the expression of TRAF4 in clinical specimens of colorectal cancer and the adjacent tissues. The expression levels of epithelial-mesenchymal transition (EMT)-related proteins [E-cadherin (E-cad), Snail and Slug] were detected by Western blotting. The Transwell assay was used to examine the invasive ability and migration ability of tumor cells LoVo after silencing TRAF4. Results Immunohistochemistry showed that TRAF4 expression was significantly higher in cancer tissues than in adjacent tissues (t=8.322, P<0.01). After transfection with TRAF4 small interfering RNA (siRNA), the expression of TRAF4 was significantly decreased (t=7.687, P<0.01), the expression of E-cad was increased (t=4.322, P<0.05), the expression of Snail and Slug was decreased (t=9.831, P<0.01, t=8.677, P<0.01), and the invasion and migration ability was obviously decreased (t=9.553, P<0.01, t=11.290, P<0.01). Conclusion Silencing TRAF4 can inhibit the invasion and migration of colorectal cancer cell LoVo. Key words: Colorectal cancer; Tumor necrosis factor receptor associated factor 4; Invasion; Migration; Epithelial-mesenchymal transition

MicroRNA-520b inhibits pancreatic cancer cell invasion and promotes apoptosis by targeting phosphofructokinase liver type

Objective To investigate the mechanism of microRNA (miRNA, miR)-520b targeting the regulation of hepatic phosphofructokinase liver type (PFKL) in pancreatic cancer cell invasion and apoptosis. Methods Cells were divided into blank group, negative control group, miR-520b positive group, miR-520b inhibition group and siRNA-PFKL group, and repeat 3 times per group. Real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) was used to detect the relative expression of miR-520b and PFKL genes. Western blotting was used to detect the levels of PFKL and epithelial-mesenchymal transition-related molecular proteins. Cell in vitro invasion assay (Transwell) and flow cytometry were used to detect cell invasion and apoptosis, respectively. Results The luciferase reporter gene assay showed that the luciferase activity of the miR-520b positive group (0.25±0.04) was significantly lower than that of the negative control group (0.92±0.09) in wild-type PFKL (P 0.05). Compared with the blank group [PFKL: 1.72±0.13, N-cadherin: 1.36±0.10, E-cadherin: 1.27±0.09, β-catenin: 0.36±0.04, Snail: 1.35±0.12, invasion: (23.27±3.18)/HF, apoptosis: (9.89±0.72)%] and the negative control group [PFKL: 1.61±0.12, N-cadherin: 1.40±0.11, E-cadherin: 1.26±0.08, β-catenin: 0.38±0.05, Snail: 1.30±0.10, invasion: (21.92±3.65)/HF, apoptosis: (10.67±0.69)%], the levels of E-cadherin protein and apoptosis were significantly increased in the miR-520b positive group [PFKL: 0.12±0.01, N-cadherin: 0.30±0.03, E-cadherin: 2.42±0.21, β-catenin: 0.19±0.02, Snail: 0.40±0.07, invasion: (12.13±1.39)/HF, apoptosis: (25.83±2.15)%] and the siRNA-PFKL group [PFKL: 0.11±0.01, N-cadherin: 0.25±0.02, E-cadherin: 2.53±0.24, β-catenin: 0.26±0.05, Snail: 0.22±0.04, invasion: (13.91±1.10)/HF, apoptosis: (23.91±1.99)%], and the levels of PFKL, N-cadherin, β-catenin, Snail protein and cell invasion were significantly reduced (P<0.05). However, the level of E-cadherin protein and apoptosis in the miR-520b inhibition group [PFKL: 3.47±0.25, N-cadherin: 3.39±0.20, E-cadherin: 0.49±0.03, β-catenin: 2.11±0.15, Snail: 2.05±0.22、invasion: (43.22±7.85)/HF, apoptosis: (2.41±0.18)%] was significantly decreased, and the levels of PFKL, N-cadherin, β-catenin, Snail protein and cell invasion were significantly increased (P<0.05). Conclusion MiR-520b can inhibit the invasion of pancreatic cancer cells and promote its apoptosis, and its mechanism is closely related to the regulation of target gene PFKL. Key words: Pancreatic cancer; MicroRNA; Invasion; Apoptosis; Phosphofructokinase liver type

Effect on invasion and migration of migration of mitofusion-2 overexpression to phosphatidylinositol transfer membrane-associated protein 3-positive hepatoma cell Line SMMC-7721

Objective To investigate the effect on invasion and migration of mitofusion-2 (Mfn-2) overexpression to phosphatidylinositol transfer membrane-associated protein 3 (PITPNM3)-positive hepatoma cell Line SMMC-7721. Methods PITPNM3 positive liver cancer cell line was selected for experiment while PITPNM3 positive breast cancer cell line McF-7 was used as a positive control group. 20 μg/L CC-chemokine ligand 18 (CCL18) was used to establish a PITPNM3 positive hepatoma cell model with invasion and migration. PITPNM3 positive hepatoma cell were transfected Mfn-2 with an adenoviral vector after 36 h to detect the change of PITPNM3 expression and its effect on invasion and migration of hepatoma cells. Results Compared to other groups, PITPNM3 was highly expressing in hepatoma cell line SMMC-7721, the relative expression level of PITPNM3 is 3.4 in SMMC-7721, 1.6 in HepG2 and 4.5 in McF-7 (SMMC-7721 vs. HepG2, t=3.653, P 0.05); 20 μg/L CCL18 can effectively promote invasion and migration of hepatoma cell line SMMC-7721 with PITPNM3 high expression, by suppressing the expression of E-Cadherin and up-regulating the expressions of Snail and Vimentin; Overexpression of Mfn-2 leads to a significant decrease in PITPNM3 expression and suppression on invasion and migration in hepatoma cell line SMMC-7721, but has no effects on the mRNA level of PITPNM3. The relative expression level of PITPNM3 is 0.903 in experimental group and 1.128 in positive control group (experimental group vs. positive control group, P>0.05). Conclusion Mfn-2 can effectively down-regulate the expression of PITPNM3 and inhibit the invasion and migration of hepatoma cell line SMMC-7721. Key words: Mitofusin-2; Phosphatidylinositol transfer membrane-associated protein 3; Hepatocellular carcinoma; Invasion; Migration

Study on the mechanism of interleukine-11 promoting anaplastic thyroid carcinoma cells invasion and metastasis

Objective To explore the mechanism of hypoxia inducing interleukine (IL)-11 secretion in anaplastic thyroid carcinoma (ATC) cells and the mechanism of IL-11 promoting epithelial mesenchymal transition of ATC cells. Methods Knockdown of IL-11, hypoxia-inducible factor (HIF)-1α in ATC cells or treatment of ATC cells with Cobalt Chloride, recombinant human interleukin (rhIL)-11, IL-11 antibody, then the effects of these interventions on the invasion and migration ability of ATC cells and its mechanism were detected by enzyme-linked immunosorbent assay (ELISA), Transwell, Real-time polymerase chain reaction (PCR), Western blotting. Results Under treatment with Cobalt Chloride, the protein and mRNA expression of IL-11 increased in ATC cells. The number of invasive cells in the knockdown HIF-1α group (61.7±4.9, 30.0±4.6) was less than that of the control group 261.7±8.7 (P<0.01), and the number of migrated cells (87.3±4.5, 73.8±5.4) was less than the control group 337.3±7.1 (P<0.01). The expression ofphosphorylated Akt (p-Akt), phosphorylated glycogen synthase kinase 3β (p-GSK3β) and Snail protein was significantly increased after treatment with rhIL-11. The protein expression of p-Akt, p-GSK3β and Snail were significantly decreased after knockdown of IL-11 expression in ATC cells. Conclusion Hypoxia induced epithelial mesenchymal transition of ATC cells is the main mechanism of high invasion and migration of ATC cells. Key words: Anaplastic thyroid carcinoma; Interleukine-11; Epithelialmesenchymal transition; Invasion and metastasis

Expression of lysine-specific demethylase 1 in hepatocellular carcinoma tissues and its effect on cell proliferation and invasion

Objective To investigate the expression of lysine-specific demethylase 1 (LSD1) in hepatocellular carcinoma tissues, and to explore the effects of down-regulation of LSD1 gene expression on proliferation and invasion of human hepatoma HepG2 cells. Methods A total of 91 cases of hepatocellular carcinoma undergoing elective surgery in our hospital and Henan Cancer Hospital were selected. Immunohistochemistry was used to detect the expression of LSD1 protein in hepatocellular carcinoma tissues and adjacent tissues. Human hepatoma HepG2 cells were cultured and divided into LSD1 interference sequence group, negative control group and blank group. Real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) was used to detect the expression of LSD1 gene in cells. Methyl thiazol tetrazolium (MTT) assay was used to detect cell proliferation. Transwell chamber was used to detect cell invasion. Western blotting was used to detect the expression of LSD1, Snail, E-cadherin and N-cadherin proteins in cells. Results The positive expression rate of LSD1 protein in hepatocellular carcinoma tissues was 81.32%, which was significantly higher than that in adjacent tissues (35.16%, P<0.01). The expression of LSD1 protein in hepatocellular carcinoma tissues was related to the degree of differentiation, TNM staging and vascular invasion (P<0.05). The relative expression level of LSD1 mRNA in LSD1 interference sequence group was 1.19±0.10, which was significantly lower than that in the negative control group and blank group (2.29±0.14 and 2.34±0.11, P<0.05). The absorbance (A) values at 24, 48, 72 and 96 h in the LSD1 interference sequence group were 0.23±0.05, 0.32±0.08, 0.38±0.08 and 0.52±0.03, respectively, which were significantly lower than those in the negative control group (0.38±0.08, 0.49±0.05, 0.67±0.04 and 0.77±0.10) and blank group (0.33±0.03, 0.52±0.09, 0.70±0.08 and 0.78±0.07, P<0.05). The number of invasive cells in the LSD1 interference sequence group was (86.3±3.9), which was significantly less than that in the negative control group (110.6±9.4) and the blank group (117.7±7.0, P<0.05). The relative expression levels of LSD1, Snail, and N-cadherin proteins in the LSD1 interference sequence group were (0.25±0.03, 0.30±0.02 and 0.32±0.03), which were significantly lower than those in the negative control group (0.77±0.06, 0.76±0.10 and 0.67±0.04) and blank group (0.84±0.14, 0.74±0.07 and 0.69±0.06, P<0.05), and the relative expression level of E-cadherin protein (0.71±0.05) was significantly higher than in the negative control group (0.37±0.02) and blank group (0.34±0.04, P<0.05). Conclusion LSD1 protein was highly expressed in hepatocellular carcinoma tissues, and might play an important role in the proliferation and invasion of human hepatoma HepG2 cells through epithelial-mesenchymal transition. Key words: Carcinoma, hepatocellular; Lysine-specific demethylase 1; Proliferation; Invasion; Epithelial-mesenchymal transition

Influence of physicochemical parameters on the spatial distribution of snail species that are intermediate hosts of human schistosomes in the Senegal River Delta.

The objective of this work was to identify possible correlations between physicochemical parameters (water temperature, water flow velocity, electrical conductivity, dissolved oxygen, salinity, pH, nitrates, and phosphates) and the spatial distribution in the Senegal River delta of snail species that are intermediate hosts of human schistosomes. Eight water points (ME1 to ME4, NE1 and NE2, TA1 and TA2) were selected in the villages of Menguègne Boye, Ndellé Boye, and Thilla for biweekly monitoring of these snails and the physicochemical parameters of the water. The results show that the spatial distribution of the snail populations is related to pH, dissolved oxygen (mg/l), conductivity, temperature (̊C), and water flow velocity (m/s).

High-mobility Group Box 1 Induces the Epithelial-mesenchymal Transition, Glycolytic Switch, and Mitochondrial Repression via Snail Activation

High-mobility Group Box 1 Induces the Epithelial-mesenchymal Transition, Glycolytic Switch, and Mitochondrial Repression via Snail Activation

The inflammatory cytokine TNF contributes with RAC3-induced malignant transformation.

RAC3 is a coactivator of steroid receptors and NF-κB. It is usually overexpressed in several tumors, contributes to maintain cancer stem cells and also to induce them when is overexpressed in non-tumoral cells. In this work, we investigated whether the inflammatory cytokine TNF may contribute to the transforming effects of RAC3 overexpression in the non-tumoral HEK293 cell line. The study model included the HEK293 tumoral transformed cell line constitutively overexpressing RAC3 by stable transfection and control non-tumoral cells transfected with an empty vector. The HeLa and T47D tumoral cells that naturally overexpress RAC3 were used as positive control.We found that TNF potentiated RAC3-induced mesenchymal transition, involving an increased E-Cadherin downregulation, Vimentin and SNAIL upregulation and enhanced migratory behavior. Moreover, concerning the molecular mechanisms by which TNF potentiates the RAC3 transforming action, they involve the IKK activation, which in addition induced the β-Catenin transactivation. Our results demonstrate that although RAC3 overexpression could be a signal strong enough to induce cancer stem cells, the inflammatory microenvironment may be playing a key role contributing to the migratory and invasive phenotype required for metastasis and cancer persistence.

Clinical study on expressions of transcription factor Snail and its related proteins in non-small cell lung cancer

Objective To detect the expressions of transcription factor Snail, E-cadherin (E-cad) and N-cadherin (N-cad) in non-small cell lung cancer (NSCLC), and explore their correlations. Methods The expression patterns of Snail, E-cad and N-cad in human lung tissues were detected by immunohistochemistry, from 59 untreated NSCLC patients in Affiliated Cancer Hospital of Xinjiang Medical University from January 2014 to December 2015 for whom clinical data were available, with adjacent tissues and 31 benign lesions as control. The expression patterns of Snail, E-cad, N-cad and clinicopathological factors were analyzed. Results The positive rate of Snail and N-cad in NSCLC was significantly higher than those in the control tissues [81.4% (48/59) vs. 37.3% (22/59) vs. 12.9% (4/31), χ2 = 9.782, P < 0.05; 76.3% (45/59) vs. 33.9% (20/59) vs. 12.9% (4/31), χ2 = 7.445, P < 0.05]. The positive rate of E-cad in NSCLC was significantly lower than those in the control tissues [20.3% (12/59) vs. 55.9% (33/59) vs. 93.5% (29/31), χ2 = 45.107, P < 0.01]. The expressions of Snail and E-cad were related to lymph node metastasis and TNM stage [snail: 97.5% (39/40) vs. 47.4% (9/19), χ2 = 7.734, P < 0.05; 12.5% (1/8) vs. 81.0% (17/21) vs. 100.0% (30/30), χ2 = 7.723, P < 0.05; E-cad: 95.0% (38/40) vs. 36.8% (7/19), χ2 = 7.190, P < 0.05; 100.0% (8/8) vs. 19.0% (4/21) vs. 0 (0/30), χ2 = 12.311, P < 0.01]. The expression of N-cad was related to lymph node metastasis, histological grade and TNM stage [97.5% (39/40) vs. 47.4% (9/19), χ2 = 11.231, P < 0.01; 28.6% (2/7) vs. 63.6% (14/22) vs. 96.7% (29/30), χ2 = 9.564, P < 0.05; 12.5% (1/8) vs. 85.7% (18/21) vs. 86.7% (26/30), χ2 = 11.664, P < 0.01]. The positive correlation was found between Snail and N-cad (r = 0.373, P < 0.01), the negative correlation was found between the expression of Snail and E-cad, N-cad and E-cad (r = -0.667, P = 0.00; r = -0.297, P = 0.01). Conclusion Snail, E-cad and N-cad are involved in the development, progression and metastasis of NSCLC, and combined detection can predict tumor invasion, metastasis and prognosis. Key words: Carcinoma, non-small-cell lung; Snail; E-cadherin; N-cadherin

Inhibitory effect of nigericin on human epithelial ovarian cancer cells and its mechanism

Objective To ascertain the specific activities of nigericin on inhibiting human epithelial ovarian cancer (EOC) cells, and to investigate the possible molecular mechanism of nigericin on cell migration and invasion. Methods Cell viability under different treatments of nigericin (0.312 5, 0.625, 1.25, 2.5, 5, 10, 20, 40, 80 μmol/L) on EOC A2780 and SKOV3 cell lines was examined by CCK-8 assay, with DMSO as a control. The human epithelial ovarian cancer cell lines A2780 and SKOV3 were treated with 5, 10 or 20 μmol/L nigericin or with DMSO as a control. Transwell chambers was used to observe the impact of nigericin on migration and invasion of EOC cells. Western blot was used to detect the expressions of epithelial cell marker (E-cadherin), mesenchymal cell marker (Vimentin) and the epithelial-mesenchymal transition (EMT)-related transcription factors Slug, Snail, and Twist, as well as the expressions of proteins related to Wnt/β-catenin signaling pathway such as Gsk-3β, p-Gsk-3β and β-catenin under different concentration treatment of nigericin. Results CCK-8 assay showed that nigericin exhibited strong cytotoxicity on A2780 [IC50 (16.19±0.26) μmol/L, 95% CI 1.077-1.341)] and SKOV3 [IC50 (11.87±0.21) μmol/L, 95% CI 1.003-1.146] cell lines. Transwell chamber assay revealed that nigericin at different concentration (5, 10, 20 μmol/L) induced a remarkable reduction in the number of cells migrating through the membrane relative to the vehicle-treated controls in A2780 and SKOV3 cells [(121±9), (92±7), (59±5)/HP and (120.4±2.6), (91.8±5.5), (80.0±4.0)/HP, all P < 0.05]; the invasive ability of A2780 and SKOV3 cells also inhibited [(61.2±3.7), (43.2±4.3), (23.6±2.1)/HP and (85.2±7.0), (65.2±4.6), (45.6±4.4)/HP, all P < 0.05]. Western blot revealed that the increased expression of E-cadherin and decreased expression of Vimentin, Slug, Snail, Twist, p-Gsk-3β, β-catenin in EOC cells with the nigericin treatment at different concentration (5, 10 and 20 μmol/L) showed concentration dependence (P < 0.05). Conclusion Nigericin may induce EMT by activating Wnt-β-catenin signaling pathway to promote the migration and invasion of EOC cells. Key words: Ovarian neoplasms; Nigericin; Epithelial-mesenchymal transition; Migration; Invasion

The role of estrogen receptor α and metastasis related gene 3 in human pancreatic cancer

Objective To investigate the role of estrogen receptor α (ERα) and metastasis related gene 3 (MTA3) in epithelial-mesenchymal transition (EMT) via ERα→MTA3→Snail→E-cadherin signaling pathway and the correlation with cell apoptosis in pancreatic duct adenocarcinoma (PDAC) cells. Methods The expression of ERα MTA3 and EMT related factors Snail and E-cadherin in human PDAC tissues was detected by immunohistochemistry, and the results were further proved in human PDAC cell lines by Western blotting. Results Most of the ERα expression was detected in cytoplasm of PDAC cells. The expression of ERα had no correlation with tumor differentiation and patients’ gender. MTA3 was found in 50% of PDAC patients, Snail in 60% and E-cadherin in 80% respectively. More expression of MTA3 was found in patients without lymph node metastasis (PP: 1.39, IRS: 2.67 vs. PP: 0.81, IRS: 2.06). Snail in low-differentiated (PP: 1.61, IRS: 3.5) was found higher than low-moderate-differentiated (PP: 1.54, IRS: 2.69) and moderate-differentiated (PP: 1.40, IRS: 2.67) tissues. A significantly more expression of E-cadherin was found in better-differentiated PDAC tissues (P=0.026). The relationship of ERα→MTA3→Snail→E-cadherin signaling pathway was found in PDAC tissues, and MTA3 expression was correlated with less Snail and more E-cadherin. Further experiment in pancreatic cell lines had the same results as in human PDAC tissues. Conclusion The factors involved in ERα→MTA3→Snail→E-cadherin signaling pathway existed in pancreatic cancer tissues, and this pathway existed in a subgroup of PDAC tissues. MTA3 is an upper stream regulator of EMT, and correlated with differentiation of PADC cells. Key words: Pancreatic adenocarcinoma; Estrogen receptor α; Metastasis related gene 3; Epithelial-mesenchymal transition

High Fecundity, Rapid Development and Selfing Ability in Three Species of Viviparous Land Snails Phaedusinae (Gastropoda: Stylommatophora: Clausiliidae) from East Asia.

Anna Sulikowska-Drozd, Takahiro Hirano, Shu-Ping Wu, and Barna Páll-Gergely (2018) Life history traits are important yet understudied aspects of ecological diversification in land snail faunas. To acquire information for comparative analysis of gastropod life cycles, we conducted experimental breeding of three viviparous clausiliids from Japan and Taiwan. Under laboratory conditions, Tauphaedusa sheridani (Pfeiffer, 1866), T. tau (O. Boettger, 1877) and Stereophaedusa (Breviphaedusa) jacobiana (Pilsbry, 1902) featured similar times to complete shell growth (12-16 weeks), age of first reproduction (23-24 weeks) and annual fecundity (143- 173 neonates per pair of snails). The maximum number of eggs retained in genital tract reaches nine for T. sheridani, 11 for T. tau, and seven for S. jacobiana. The ratio between average shell height of neonate and adult varied between 13.5% for T. tau, 13.9% for T. sheridani, and 16.7% for S. jacobiana. All species were able to self-fertilize. Exposure to long drought affects intrauterine embryo development in all species; additionally, it results in parent and embryo mortality in S. jacobiana. Observed differences in the structure of the egg envelope among studied species point to greater than expected differentiation of developmental processes in viviparous Phaedusinae of East Asia.