Variations in soil physico-chemical properties, soil stocks, and soil stoichiometry under different soil layers, the major forest region Liupan Mountains of Northwest China

Abstract Liupan Mountains are an important region in China in the context of forest cover and vegetation due to huge afforestation and plantation practices, which brought changes in soil physio-chemical properties, soil stocks, and soil stoichiometries are rarely been understood. The study aims to explore the distribution of soil nutrients at 1-m soil depth in the plantation forest region. The soil samples at five depth increments (0-20, 20-40, 40-60, 60-80, and 80-100 cm) were collected and analyzed for different soil physio-chemical characteristics. The results showed a significant variation in soil bulk density (BD), soil porosity, pH, cation exchange capacity (CEC), and electric conductivity (EC) values. More soil BD (1.41 g cm-3) and pH (6.97) were noticed in the deep soil layer (80-100 cm), while the highest values of porosity (60.6%), EC (0.09 mS cm-1), and CEC (32.9 c mol kg-1) were reflected in the uppermost soil layer (0-20 cm). Similarly, the highest contents of soil organic carbon (SOC), total phosphorus (TP), available phosphorus (AP), total nitrogen (TN), and available potassium (AK) were calculated in the surface soil layer (0-20 cm). With increasing soil depth increment a decreasing trend in the SOC and other nutrient concentration were found, whereas the soil total potassium (TK) produced a negative correlation with soil layer depth. The entire results produced the distribution of SOCs and TNs (stocks) at various soil depths in forestland patterns were 020cm > 2040cm > 4060cm 6080cm 80100 cm. Furthermore, the stoichiometric ratios of C, N, and P, the C/P, and N/P ratios showed maximum values (66.49 and 5.46) in 0-20 cm and lowest values (23.78 and 1.91) in 80-100 cm soil layer depth. Though the C/N ratio was statistically similar across the whole soil profile (0-100 cm). These results highlighted that the soil depth increments might largely be attributed to fluctuations in soil physio-chemical properties, soil stocks, and soil stoichiometries. Further study is needed to draw more conclusions on nutrient dynamics, soil stocks, and soil stoichiometry in these forests.

Resumo As montanhas de Liupan são uma região importante na China no contexto de cobertura florestal e vegetação devido às enormes práticas de florestamento e plantação, que trouxeram mudanças nas propriedades físico-químicas do solo, e estoques e estequiometrias do solo raramente são compreendidos. O estudo visa explorar a distribuição de nutrientes do solo a 1 m de profundidade do solo na região da floresta plantada. As amostras de solo em cinco incrementos de profundidade (0-20, 20-40, 40-60, 60-80 e 80-100 cm) foram coletadas e analisadas para diferentes características físico-químicas do solo. Os resultados mostraram uma variação significativa nos valores de densidade do solo (BD), porosidade do solo, pH, capacidade de troca catiônica (CEC) e condutividade elétrica (CE). Mais DB do solo (1,41 g cm-3) e pH (6,97) do solo foram observados na camada profunda do solo (80-100 cm), enquanto os maiores valores de porosidade (60,6%), CE (0,09 mS cm-1) e CEC (32,9 c mol kg-1) foram refletidos na camada superior do solo (0-20 cm). Da mesma forma, os maiores teores de carbono orgânico do solo (SOC), fósforo total (TP), fósforo disponível (AP), nitrogênio total (TN) e potássio disponível (AK) foram calculados na camada superficial do solo (0-20 cm). Com o aumento do incremento da profundidade do solo, uma tendência decrescente no SOC e na concentração de outros nutrientes foi encontrada, enquanto o potássio total do solo (TK) produziu uma correlação negativa com a profundidade da camada do solo. Todos os resultados produziram a distribuição de SOCs e TNs (estoques) em várias profundidades de solo em padrões de floresta 0 20cm> 20 40cm> 40 60cm 60 80cm 80 100 cm. Além disso, as relações estequiométricas de C, N e P, as relações C / P e N / P, apresentaram valores máximos (66,49 e 5,46) em 0-20 cm, e valores mais baixos (23,78 e 1,91) em solo de 80-100 cm profundidade da camada. Embora a relação C / N fosse estatisticamente semelhante em todo o perfil do solo (0-100 cm). Esses resultados destacaram que os incrementos de profundidade do solo podem ser amplamente atribuídos a flutuações nas propriedades físico-químicas do solo, estoques e estequiometrias do solo. Mais estudos são necessários para tirar conclusões adicionais sobre a dinâmica dos nutrientes, estoques de solo e estequiometria do solo nessas florestas.

Variations in soil physico-chemical properties, soil stocks, and soil stoichiometry under different soil layers, the major forest region Liupan Mountains of Northwest China / Variações nas propriedades físico-químicas do solo, estoques e estequiometria do solo sob diferentes camadas de solo, a principal região florestal das montanhas Liupan do noroeste da China

Liupan Mountains are an important region in China in the context of forest cover and vegetation due to huge afforestation and plantation practices, which brought changes in soil physio-chemical properties, soil stocks, and soil stoichiometries are rarely been understood. The study aims to explore the distribution of soil nutrients at 1-m soil depth in the plantation forest region. The soil samples at five depth increments (0-20, 20-40, 40-60, 60-80, and 80-100 cm) were collected and analyzed for different soil physio-chemical characteristics. The results showed a significant variation in soil bulk density (BD), soil porosity, pH, cation exchange capacity (CEC), and electric conductivity (EC) values. More soil BD (1.41 g cm-3) and pH (6.97) were noticed in the deep soil layer (80-100 cm), while the highest values of porosity (60.6%), EC (0.09 mS cm-1), and CEC (32.9 c mol kg-1) were reflected in the uppermost soil layer (0-20 cm). Similarly, the highest contents of soil organic carbon (SOC), total phosphorus (TP), available phosphorus (AP), total nitrogen (TN), and available potassium (AK) were calculated in the surface soil layer (0-20 cm). With increasing soil depth increment a decreasing trend in the SOC and other nutrient concentration were found, whereas the soil total potassium (TK) produced a negative correlation with soil layer depth. The entire results produced the distribution of SOCs and TNs (stocks) at various soil depths in forestland patterns were 0→20cm > 20→40cm > 40→60cm ≥ 60→80cm ≥ 80→100 cm. Furthermore, the stoichiometric ratios of C, N, and P, the C/P, and N/P ratios showed maximum values (66.49 and 5.46) in 0-20 cm and lowest values (23.78 and 1.91) in 80-100 cm soil layer depth. Though the C/N ratio was statistically similar across the whole soil profile (0-100 cm). These results highlighted that the soil depth increments might largely be attributed to fluctuations in soil physio-chemical properties, soil stocks, and soil stoichiometries. Further study is needed to draw more conclusions on nutrient dynamics, soil stocks, and soil stoichiometry in these forests.

As montanhas de Liupan são uma região importante na China no contexto de cobertura florestal e vegetação devido às enormes práticas de florestamento e plantação, que trouxeram mudanças nas propriedades físico-químicas do solo, e estoques e estequiometrias do solo raramente são compreendidos. O estudo visa explorar a distribuição de nutrientes do solo a 1 m de profundidade do solo na região da floresta plantada. As amostras de solo em cinco incrementos de profundidade (0-20, 20-40, 40-60, 60-80 e 80-100 cm) foram coletadas e analisadas para diferentes características físico-químicas do solo. Os resultados mostraram uma variação significativa nos valores de densidade do solo (BD), porosidade do solo, pH, capacidade de troca catiônica (CEC) e condutividade elétrica (CE). Mais DB do solo (1,41 g cm-3) e pH (6,97) do solo foram observados na camada profunda do solo (80-100 cm), enquanto os maiores valores de porosidade (60,6%), CE (0,09 mS cm-1) e CEC (32,9 c mol kg-1) foram refletidos na camada superior do solo (0-20 cm). Da mesma forma, os maiores teores de carbono orgânico do solo (SOC), fósforo total (TP), fósforo disponível (AP), nitrogênio total (TN) e potássio disponível (AK) foram calculados na camada superficial do solo (0-20 cm). Com o aumento do incremento da profundidade do solo, uma tendência decrescente no SOC e na concentração de outros nutrientes foi encontrada, enquanto o potássio total do solo (TK) produziu uma correlação negativa com a profundidade da camada do solo. Todos os resultados produziram a distribuição de SOCs e TNs (estoques) em várias profundidades de solo em padrões de floresta 0 → 20cm> 20 → 40cm> 40 → 60cm ≥ 60 → 80cm ≥ 80 → 100 cm. Além disso, as relações estequiométricas de C, N e P, as relações C / P e N / P, apresentaram valores máximos (66,49 e 5,46) em 0-20 cm, e valores mais baixos (23,78 e 1,91) em solo de 80-100 cm profundidade da camada. Embora a relação C / N fosse estatisticamente semelhante em todo o perfil do solo (0-100 cm). Esses resultados destacaram que os incrementos de profundidade do solo podem ser amplamente atribuídos a flutuações nas propriedades físico-químicas do solo, estoques e estequiometrias do solo. Mais estudos são necessários para tirar conclusões adicionais sobre a dinâmica dos nutrientes, estoques de solo e estequiometria do solo nessas florestas.

Culture-negative TB: clinical characteristics, risk factors and treatment outcomes.

BACKGROUND: Although culture remains the standard for TB diagnosis, 15-20% of patients diagnosed and treated for TB are culture-negative. We explored clinical characteristics, risk factors and treatment outcomes for culture-negative TB in a Peruvian cohort.METHODS: We recruited 4,500 index TB patients and 10,160 household contacts in Lima, Peru, and enrolled 692 secondary patients diagnosed with TB during follow-up of household contacts. We analyzed smear and culture status, sociodemographic factors, clinical characteristics and TB treatment outcomes to compare culture-negative and positive patients.RESULTS: Of the 4,880 adult patients, 915 (18.8%) were culture-negative. Culture-negative patients were less likely to report symptoms of TB disease and disease of longer duration. A multivariate analysis showed no statistically significant difference in loss to follow-up, treatment failure or recurrence between the culture-negative and -positive groups but a higher rate of death among culture-negative patients with an adjusted OR of 1.65 (95% CI 1.05-2.60). In a multivariate analysis of determinants of culture negativity, older age, substance use and being a secondary case were associated with culture status.CONCLUSIONS: More recognition and awareness of culture-negative TB is key for early and correct diagnosis to reduce transmission and improve treatment outcomes.

The Effects of Yucca Schidigera Extract and Multi-carbohydrase in Different Crude Protein Diets on Growth Performance, Nutrient Digestibility, Carcass Parameters and Excreta Noxious Gas Contents in Broilers

The scarcity and increase in the price of animal feeds have attracted the attention of nutritionists to address this issue. The inclusion of plant extracts and enzymes to protein-reduced diet could be a feasible strategy to reducing the feed cost. Therefore, the purpose of this study was to assess the impacts of Yucca schidigera extract and multi-carbohydrase in low crude protein (CP) diets of broiler on growth performance, nutrient digestibility, carcass metrics, and noxious gas levels in the excreta. A total of 480, 1-d-old ROSS 308 were randomly allocated into 4 dietary treatments, six replication, and 20 birds/cage. Phase 1, T1(CP 21%, ME 2,969 kcal/kg); T2 (CP 19%, ME 2,863 kcal/kg + 0.02% Yucca); T3 (CP 17%, ME 2,865 kcal/kg + 0.02% Yucca); T4 (CP 17%, ME 2,861 kcal/kg + 0.02% Yucca + 0.1% multi-carbohydrase). Phase 2, T1 (CP 19%, ME 3,086 kcal/kg); T2 (CP 17%, ME 2,977 kcal/kg + 0.02% Yucca); T3 (CP 15%, ME 2,978 kcal/kg + 0.02% Yucca); T4 (CP 15%, ME 2,978 kcal/kg + 0.02% Yucca + 0.1% 0.1% multi-carbohydrase). Although the addition of YS and multi-carbohydrase to the low CP diets on the growth performance did not improve, it revealed the positive result on the nutrient digestibility, carcass parameters, and noxious gas emission. Overall, broilers supplemented with YS 0.02% and multi-carbohydrase (0.1%) demonstrated the best production performances compared to the other treatment groups. Thus, a combination of YS and multi-carbohydrase could be added to the diets with low CP to boost broiler production performance.(AU)

Comparison of Characteristics and Differences in Early Immune Organ Development in Different Strains of Tianfu Broiler

To effectively develop and utilize high-quality Tianfu broilers, this study evaluated the morphological and structural characteristics of the immune organs of such broilers with different strains (HS1 and HS2) at different developmental stages and analyzed the distribution of mast cells by toluidine blue staining. Moreover, the localization and expression of immunoglobulin, complement C3, C4 and CD3 in immune organs were also detected. The results showed that although there was no significant difference in the development of immune organs in the HS1 and HS2, the number of lymphatic follicles and capsule thickness in the spleen and bursa of Fabricius in HS1 were greater than those in HS2. Additionally, the number of mast cells in the spleen of HS1 was greater at Day 1 and Day 21 and was significantly higher than that of HS2 (p<0.05); the number of mast cells in the bursa of Fabricius reached 9.17 on Day 7, which was significantly higher than that of HS2 (p<0.05). Moreover, the serum IgA and IgM levels in HS1 were higher than those in HS2 on Day 14 and 21 (p<0.05). In addition, the complement C3 content in HS1 was significantly or extremely significantly higher than that in HS2 on Days 1, 14 and 21 (p<0.01, p<0.05), respectively, but significantly lower than in HS2 on Day 7 (p<0.05). These results indicated that the disease resistance of the HS1 line was stronger than that of the HS2 line, which lays a foundation for future disease- resistance breeding of Tianfu broilers.(AU)

Variations in soil physico-chemical properties, soil stocks, and soil stoichiometry under different soil layers, the major forest region Liupan Mountains of Northwest China.

Liupan Mountains are an important region in China in the context of forest cover and vegetation due to huge afforestation and plantation practices, which brought changes in soil physio-chemical properties, soil stocks, and soil stoichiometries are rarely been understood. The study aims to explore the distribution of soil nutrients at 1-m soil depth in the plantation forest region. The soil samples at five depth increments (0-20, 20-40, 40-60, 60-80, and 80-100 cm) were collected and analyzed for different soil physio-chemical characteristics. The results showed a significant variation in soil bulk density (BD), soil porosity, pH, cation exchange capacity (CEC), and electric conductivity (EC) values. More soil BD (1.41 g cm-3) and pH (6.97) were noticed in the deep soil layer (80-100 cm), while the highest values of porosity (60.6%), EC (0.09 mS cm-1), and CEC (32.9 c mol kg-1) were reflected in the uppermost soil layer (0-20 cm). Similarly, the highest contents of soil organic carbon (SOC), total phosphorus (TP), available phosphorus (AP), total nitrogen (TN), and available potassium (AK) were calculated in the surface soil layer (0-20 cm). With increasing soil depth increment a decreasing trend in the SOC and other nutrient concentration were found, whereas the soil total potassium (TK) produced a negative correlation with soil layer depth. The entire results produced the distribution of SOCs and TNs (stocks) at various soil depths in forestland patterns were 0→20cm > 20→40cm > 40→60cm ≥ 60→80cm ≥ 80→100 cm. Furthermore, the stoichiometric ratios of C, N, and P, the C/P, and N/P ratios showed maximum values (66.49 and 5.46) in 0-20 cm and lowest values (23.78 and 1.91) in 80-100 cm soil layer depth. Though the C/N ratio was statistically similar across the whole soil profile (0-100 cm). These results highlighted that the soil depth increments might largely be attributed to fluctuations in soil physio-chemical properties, soil stocks, and soil stoichiometries. Further study is needed to draw more conclusions on nutrient dynamics, soil stocks, and soil stoichiometry in these forests.

Bone marrow mesenchymal stem cells promote the progression of prostate cancer through the SDF-1/CXCR4 axis in vivo and vitro.

PURPOSE: The aim of this study was to investigate the involvement of the SDF-1/CXCR4 axis in the process of BMMSC homing in prostate cancer (PCa) in vivo and in vitro. METHODS: After verification of BMMSCs, we fixed the concentration gradient of SDF-1 for BMMSC cultivation to analyze CXCR4 expression by qRT-PCR and flow cytometric analysis. Furthermore, we developed a non-contact co-culture system and explored the participation of the SDF-1/CXCR4 axis in PCa using qRT-PCR, flow cytometry, and ELISA. In addition, A green fluorescent protein (GFP)-transplanted methylnitrosourea (MNU)-induced PCa mouse model was established to investigate the CXCR4 expression in vivo. RESULTS: The CXCR4 expression was up-regulated with the increase in SDF-1 concentrations, and elevated SDF-1 had a significant promoting effect on cell proliferation and migration in BMMSCs. Moreover, the CXCR4 expression of BMMSCs was significantly increased in the non-contact co-culture model with vascular endothelial cells (VECs), and analysis of this model also showed that the proliferation and migration of BMMSCs were promoted in the presence of VECs. The ELISA assay showed that the SDF-1 levels in the co-culture model at 48 h were significantly increased. Twenty of the GFP-transplanted mice were divided into a PCa group and a control group, and four GFP-transplanted mice were observed to have prostate tumorigenesis. It also showed that CXCR4 was obviously increased in the prostate tissue of PCa mice. CONCLUSION: Our findings suggest that BMMSCs could home and promote the proliferation and migration of PCa through the SDF-1/CXCR4 axis in vivo and in vitro.

Silencing IKBKE inhibits the migration and invasion of glioblastoma by promoting Snail1 degradation.

PURPOSE: Glioblastoma multiforme (GBM) is one of the most common malignant brain tumors in adults and has high mortality and relapse rates. Over the past few years, great advances have been made in the diagnosis and treatment of GBM, but unfortunately, the five-year overall survival rate of GBM patients is approximately 5.1%. Inhibitor of nuclear factor kappa-B kinase subunit epsilon (IKBKE) is a major oncogenic protein in tumors and can promote evil development of GBM. Snail1, a key inducer of the epithelial-mesenchymal transition (EMT) transcription factor, is subjected to ubiquitination and degradation, but the mechanism by which Snail1 is stabilized in tumors remains unclear. Our study aimed to investigate the mechanism of IKBKE regulating Snail1 in GBM. METHODS: First, we analyzed the correlation between the expression of IKBKE and the tumor grade and prognosis through public databases and laboratory specimen libraries. Second, immunohistochemistry (IHC) and western blot were used to detect the correlation between IKBKE and Snail expression in glioma samples and cell lines. Western blot and immunofluorescence (IF) experiments were used to detect the quality and distribution of IKBKE and Snail1 proteins. Third, In situ animal model of intracranial glioma to detect the regulatory effect of IKBKE on intracranial tumors. RESULTS: In this study, Our study reveals a new connection between IKBKE and Snail1, where IKBKE can directly bind to Snail1, translocate Snail1 into the nucleus from the cytoplasm. Downregulation of IKBKE results in Snail1 destabilization and impairs the tumor cell migration and invasion capabilities. CONCLUSION: Our studies suggest that the IKBKE-Snail1 axis may serve as a potential therapeutic target for GBM treatment.

Effects of Vitamin A and K3 on Immune Function and Intestinal Antioxidant Capacity of Aged Laying Hens

This study was conducted to investigate effects of vitamin A (VA) and vitamin K3 (VK3) on immune function and intestinal antioxidant capacity of aged laying hens. In a 3 × 3 factorial arrangement, the diets of 1080 Roman Pink laying hens (87 weeks old) was formulated with deficient, adequate and excess VA and VK3, including 0, 7000 and 14000 IU/kg VA and 0, 2.0 and 4.0 mg/kg VK3 for 8 weeks. Interactive effects between VA and VK3 were observed that VA and VK3 decreased the splenetic mRNA expression of inducible nitric oxide synthase (iNOS) and tumour necrosis factor α (TNF-α), but increased the plasma immunoglobulin G (IgG) content and jejunal mRNA expression of nuclear factor-like 2 (Nrf2). Hens fed adequate or excess VA had higher spleen index, mRNA expression of interleukin-10 (IL-10) in spleen, sIgA content, catalase (CAT), glutathione peroxidase and total dismutase (T-SOD) activity, and mRNA expression of polymeric immunoglobulin receptor (pIgR) in jejunum and lower mRNA expression of IL-1ß in jejunum and iNOS, TNF-α in spleen. Furthermore, adequate or excess VK3 significantly increased plasma IgG content, the CAT, T-SOD and total antioxidant capacity activities, up-regulated the mRNA expression of pIgR, Nrf2, SOD1 and CAT in jejunum and down-regulated the mRNA expression of iNOS and TNF-α in spleen.(AU)

Circular RNA circHIPK3 modulates prostate cancer progression via targeting miR-448/MTDH signaling.

PURPOSE: Prostate cancer (PCa) is one of the most diagnosed cancers in men worldwide. Several studies have identified that circular RNAs (circRNAs) have a crucial impact on the biological processes in PCa. Therefore, it is necessary to study the molecular mechanism of circRNAs in tumor progression and metastasis. METHODS: RNA interference was used to decrease circHIPK3 and MTDH expression. Overexpression vector was used to increase circHIPK3 and MTDH expression. Luciferase reporter assay were used to detect the relationship between circHIPK3 and miR-448 or between miR-448 and MTDH. MTT assay, colony formation assay and transwell assay were used to measure proliferation and migration of PCa cells. RESULTS: Circular RNA circHIPK3 was significantly increased in PCa tissues and cell lines. And overexpression of circHIPK3 promoted the migration, proliferation, and invasion of PC-3 and 22Rv1 cells, while knockdown of circHIPK3 markedly repressed the above-mentioned series of biological processes. Furthermore, circHIPK3 promoted metadherin (MTDH) expression by sponging miR-448. In vivo experiments, it was also found that overexpression of circHIPK3 significantly promoted tumor growth. CONCLUSIONS: Our research shows that circHIPK3 plays a carcinogenic effect in PCa by regulating the miR-448/MTDH axis, indicating that circHIPK3 may be a potential therapeutic target for PCa.

miR-144-3p increases radiosensibility of gastric cancer cells by targeting inhibition of ZEB1.

PURPOSE: This study set out to probe into the effect and mechanism of miR-144-3p on radiosensitivity of gastric cancer (GC) cells. METHODS: Cancer tissue and paracancerous tissue of GC patients admitted to our hospital were collected, their miR-144-3p expression was tested, GC cells were transfected, and survival and biological behavior of those cells under radiation were detected. RESULTS: After detection, miR-144-3p expression was down-regulated in GC tissue, while ZEB1 was up-regulated. There was no remarkable difference in the survival fraction of cells in each group before receiving radiation, but that of tumor cells decreased obviously (p < 0.05) after radiation exposure. Survival fraction of cells overexpressing miR-144-3p or silencing ZEB1 decreased more obviously, while the inhibition of miR-144-3p or overexpressing ZEB1 was weaker. Biological behavior of cells under 6 Gy radiation was detected. It was found that miR-144-3p overexpression or silencing ZEB1 dramatically inhibited the proliferation activity of GC cells under 6 Gy radiation, increased the levels of pro-apoptotic Bax and caspase-3 proteins (p < 0.05) and decreased the anti-apoptotic protein Bcl-2 level (p < 0.05), resulting in an increase in the apoptosis rate of cells. miR-144-3p was confirmed to be ZEB1 targeting site by dual luciferase report. Moreover, rescue experiments prove that it can increase the radiosensitivity of GC cells by regulating ZEB1 expression. CONCLUSION: miR-144-3p expression was down-regulated in GC, and it can increase the radiosensitivity of those cells by inhibiting ZEB1 expression.

ADMA mediates gastric cancer cell migration and invasion via Wnt/ß-catenin signaling pathway.

OBJECTIVE: To explore the role of ADMA in gastric cancer. METHODS: The specimens of 115 gastric cancer patients were analyzed by ELISA and survival analysis. Functional assays were used to assess the effects of ADMA on gastric cancer cells. Experiments were conducted to detect the signaling pathway induced by ADMA in GC. RESULTS: Gastric cancer patients with high ADMA levels had poor prognosis and low survival rate. Furthermore, high level of ADMA did not affect the proliferation while promoted the migration and invasion of gastric cancer cell. Moreover, ADMA enhanced the epithelial-mesenchymal transition (EMT). Importantly, ADMA positively regulated ß-catenin expression in GC and promoted GC migration and invasion via Wnt/ß-catenin pathway. CONCLUSIONS: ADMA regulates gastric cancer cell migration and invasion via Wnt/ß-catenin signaling pathway and which may be applied to clinical practice as a diagnostic and prognostic biomarker.

Paeonol inhibits proliferation and induces cell apoptosis of human T24 and 5637 bladder cancer cells in vitro and in vivo.

PURPOSE: Paeonol is a natural chemical medicine derived from the bark of peony root, which has been found to inhibit tumor activity in various tumor cell lines, and can play a synergistic anti-tumor effect with chemotherapy or radiotherapy. METHODS: We used paeonol to act on human bladder cancer T24 and 5637 cells, and established xenograft tumor in nude mice by subcutaneous injection of T24 cells. RESULTS: CCK-8 assay and plate cloning experiments showed that paeonol could inhibit the proliferation of T24 and 5637 cells in vitro. The results of flow cytometry and the detection of BAX, Bcl-2 and Caspase-3 proteins suggested that paeonol can induce apoptosis of T24 and 5637 cells in vitro. Tumor formation, TUNEL detection and immunohistochemical results of Ki67, BAX, Bcl-2 and Caspase-3 in nude mice showed that paeonol could inhibit T24 cell proliferation and induce apoptosis in vivo, thus inhibiting tumor growth. Further research revealed that paeonol could reduce phosphorylation expression of PI3K and AKT in T24 and 5637 cells. CONCLUSION: We confirmed that paeonol could inhibit proliferation and induce apoptosis of human bladder cancer T24 and 5637 cells in vitro and in vivo, inhibit the growth of T24 tumor-forming nude mice, and possibly play a role by inhibiting the PI3K/AKT signaling pathway, so as to provide a potential therapeutic drug for bladder cancer.

Gefitinib promotes CXCR4-dependent epithelial to mesenchymal transition via TGF-ß1 signaling pathway in lung cancer cells harboring EGFR mutation.

PURPOSE: Epithelial to mesenchymal transition (EMT) plays an important role in acquired resistance to gefitinib in lung cancer. This study aimed to explore the underlying mechanism of gefitinib-induced EMT in lung adenocarcinoma cells harboring EGFR mutation. METHODS: CXC chemokine receptor 4 (CXCR4) expression was determined through qRT-PCR, Western blot and flow cytometry assays in lung cancer cell line (PC9) bearing mutated EGFR. Functional role of CXCR4 was inhibited applying siRNAs as well as the specific antagonist AMD3100. The expression of EMT markers was determined, and the migration of PC9 cells was measured with transwell assay. RESULTS: We found that gefitinib promoted the migratory capacity of PC9 cells in vitro, which correlated with EMT occurrence through upregulation of CXCR4. Blocking CXCR4 significantly suppressed gefitinib-induced enhancement of migration and EMT. Moreover, we determined that the upregulation of CXCR4 by gefitinib was dependent on TGF-ß1/Smad2 signaling activity. CONCLUSIONS: Our study suggested a potential mechanism by which gefitinib induced EMT in cells harboring EGFR mutation through a pathway involving TGF-ß1 and CXCR4. Thus, the combination of CXCR4 antagonist and TGFßR inhibitors might provide an alternative strategy to overcome progression of lung cancer after gefitinib treatment.

Dynamic monitoring of HER2 amplification in circulating DNA of patients with metastatic colorectal cancer treated with cetuximab.

PURPOSE: Cetuximab (CTX) has been used to treat metastatic colorectal cancer (mCRC) with wild-type (wt) RAS and BRAF genes. Meanwhile HER2 amplification reportedly denoted CTX-resistant mCRC tumors. We investigated whether monitoring of HER2 amplification in circulating DNA allowed early detection of mCRC progression and CTX resistance. METHODS: We analyzed HER2 amplification in circulating DNA at 8-week intervals using ddPCR from 36 patients with RASwt/BRAFwt mCRC, who progressed after CTX treatments between July 2015 and January 2018. RESULTS: Of the 36 patients, 5 (13.9%) exhibited dynamic fluctuations of HER2 amplification in plasma in the course of CTX treatment, of whom 2 were positive for HER2 amplification in matched tumor specimens at baseline (per FISH). All 5 primary sites were left side: 3 rectums and 2 descending colon. HER2 ratio fluctuations in circulating DNA not only reflected changes in tumor volume, but their obvious increases presaged CT-documented progress by an average lead time of 2 months. Interestingly, progression-free survival did not significantly differ between these 5 patients and those without HER2 amplification (HR 1.06, 95% CI 0.40-2.77, P = 0.909). CONCLUSION: Plasma HER2 amplification detected by ddPCR changed over time and predicted resistance to CTX, by an average lead time of 2 months. Further study is needed to validate our findings.

IKBKE enhances TMZ-chemoresistance through upregulation of MGMT expression in glioblastoma.

PURPOSE: Glioblastoma multiforme (GBM) is the most common and aggressive malignant type of brain tumor. Despite advances in diagnosis and therapy, the prognosis of patients with GBM has remained dismal. Multidrug resistance and high recurrence are two of the major challenges in successfully treating brain tumors. IKBKE (inhibitor of nuclear factor kappa-B kinase subunit epsilon) is a major oncogenic protein in tumors and can inhibit glioblastoma cell proliferation, migration, and tumorigenesis. Our study aimed to investigate the mechanism of IKBKE enhancing the resistance of glioma cells to temozolomide. METHODS: For the in vitro experiments, LN18 and U118 glioblastoma cells were treated with a combination of sh/oe-IKBKE lentivirus and TMZ. Cell proliferation was determined by the EdU assay and colony formation assays. Apoptosis was analyzed by the TUNEL assay. In vivo, LN18 NC and LN18 sh-IKBKE cells were implanted into the cerebrums of nude mice to detect the effect of combination therapy. The protein and mRNA levels were assayed by western blot, immunohistochemistry, and qRT-PCR. RESULTS: In this study, we demonstrated that IKBKE enhances the resistance of glioblastoma cells to temozolomide (TMZ) by activating the AKT/NF-κB signaling pathway to upregulate the expression of the DNA repair enzyme o6-methylguanine-dna methyltransferase (MGMT). In glioblastoma cells, IKBKE knockdown enhances apoptosis and suppresses cell proliferation, clone formation, and tumor development in vivo induced by TMZ. However, overexpression of IKBKE reduces the effects of TMZ. CONCLUSION: Our studies suggest that inhibition of IKBKE can enhance the therapeutic effect of TMZ on GBM in vitro and in vivo, providing new research directions and therapeutic targets for the treatment of GBM.

Laparoscopy adjuvant total colorectal resection for the treatment of familial adenomatous polyposis (FAP).

OBJECTIVE: To discuss and evaluate the safety and value of laparoscopy adjuvant total colorectal resection for the treatment of familial adenomatous polyposis (FAP). METHODS: From March 2010 to June 2015, 38 cases were retrospectively analyzed and divided into 2 groups, of which 17 cases used laparoscopy adjuvant total colorectal resection, and 21 cases used conventional laparotomy. Clinical data were obtained, and the safety and prognosis were observed. RESULTS: Seventeen cases using laparoscopy adjuvant total colorectal resection achieved success with no conversion to laparotomy and intraoperative complications. There was no significant difference in operation time between the two groups. There were significant differences in blood loss, the length of incision, postoperative recovery time of intestinal function and postoperative hospital stay between the two groups (P < 0.05). The trauma in laparoscopy group was less, and could recover faster, and there was no significant difference in complications between the two groups. In addition, there were no recurrence, distant metastasis and death in the follow-up period from 6 to 56 months. CONCLUSION: Laparoscopy adjuvant total colorectal resection is more safe and feasible, which has minimal invasion and can recover fast.

Detection of Snps in the Melanocortin 1-Receptor (MC1R) and Its Association with Shank Color Trait in Hs Chicken

The melanocortin 1 receptor (MC1R) gene plays a key role in controlling the deposition of melanin. In mammals, the MC1Rgene is regarded as a major candidate gene in the control of melanin formation. In domestic animals, the MC1R gene mainly controls the expression of coat, skin, and plumage color in mammals and birds. In order to breed chickens with dark-green shank faster, we screened the molecular markers for shank color in a HS chicken population by exploring the relationship between polymorphism of the MC1R gene and three different shank colors (light green, dark green and yellow). Two primer pairs for code region of the MC1R gene were designed in the basic of chicken genomic sequence. DNA sequencing was performed to detect the polymorphisms and PCR was used to amplify DNA fragment. Sequences analysis indicated that 7 SNPs were predominant the three HS chicken populations with different shank color, including g.18,287,945C>T, g.18,288,088T>C, g.18,288,150G>A, g.18,288,303A>G, g.18,288,512G>A, g.18,288,513T>C, and g.18,288,520A>C. Association analysis revealed that the dark-green shank population showed moderate polymorphism, whereas the light-green shank population showed low polymorphism among overall 7 SNPs and that SNP6 (g.18,288,513T>C) may be significantly associated with three different shank colors in HS chickens. The haplotype CTGGACA had the largest haplotype frequencies, accounting for 56.22%, and the haplotype combination H1H1 is mainly distributed in the dark-green shank population, and may be used as molecular maker for marker-assisted selection of shank color in HS chickens.

Genetic Diversity and Phyletic Evolution of Eleven Chinese Indigenous and Three Commercial Chicken Breeds by mtDNA Sequences

Chinese indigenous chicken breeds are geographically widespread, and a total of 116 indigenous chicken breeds are listed as Chinese national genetic resources. However, these indigenous chicken breeds are facing serious challenges as declining population and germplasm degeneration because lots of commercial chicken breeds had been introduced. In this study, the genetic variations of eleven Chinese indigenous chicken breeds of Sichuan province and three commercial chicken breeds were investigated based on the partial mitochondrial DNA D-loop of 487bp in length. 147 individuals from 14 breeds were examined and 34 haplotypes were observed. Genetic diversity analysis showed that the highest haplotype diversity level was found in Dahen Chicken (DH) population, while the Arbor Acres Chicken (WF) and Roman layer (RM) showed lower genetic diversity levels. The long-term artificial selection may lead to reduced nucleotide diversity. Genetic population differentiation analysis indicated that most of the variation (80.80%) was attributed to variations among breeds. Phylogenetic analysis revealed that these individuals were divided into four distinct genetic clades, including cluster A, B, C and D. Eighteen haplotypes were classified as cluster A, eight haplotypes were classified as cluster B, five haplotypes were classified as cluster C and three haplotypes were classified as cluster D. There was no breed-specific clade. Our study firstly identified the populations genetic structure of Chinese indigenous chickens and the most important commercial breeds in Sichuan province, though the genetic diversity of indigenous breeds did not suffer obvious decrease, but could be helpful for efficient artificial breeding selection and genetic resources conservation.

Detection of Snps in the Melanocortin 1-Receptor (MC1R) and Its Association with Shank Color Trait in Hs Chicken

The melanocortin 1 receptor (MC1R) gene plays a key role in controlling the deposition of melanin. In mammals, the MC1Rgene is regarded as a major candidate gene in the control of melanin formation. In domestic animals, the MC1R gene mainly controls the expression of coat, skin, and plumage color in mammals and birds. In order to breed chickens with dark-green shank faster, we screened the molecular markers for shank color in a HS chicken population by exploring the relationship between polymorphism of the MC1R gene and three different shank colors (light green, dark green and yellow). Two primer pairs for code region of the MC1R gene were designed in the basic of chicken genomic sequence. DNA sequencing was performed to detect the polymorphisms and PCR was used to amplify DNA fragment. Sequences analysis indicated that 7 SNPs were predominant the three HS chicken populations with different shank color, including g.18,287,945C>T, g.18,288,088T>C, g.18,288,150G>A, g.18,288,303A>G, g.18,288,512G>A, g.18,288,513T>C, and g.18,288,520A>C. Association analysis revealed that the dark-green shank population showed moderate polymorphism, whereas the light-green shank population showed low polymorphism among overall 7 SNPs and that SNP6 (g.18,288,513T>C) may be significantly associated with three different shank colors in HS chickens. The haplotype CTGGACA had the largest haplotype frequencies, accounting for 56.22%, and the haplotype combination H1H1 is mainly distributed in the dark-green shank population, and may be used as molecular maker for marker-assisted selection of shank color in HS chickens.(AU)