Exploring the autophagy-related pathogenesis of active ulcerative colitis.

BACKGROUND: The pathogenesis of ulcerative colitis (UC) is complex, and recent therapeutic advances remain unable to fully alleviate the condition. AIM: To inform the development of novel UC treatments, bioinformatics was used to explore the autophagy-related pathogenesis associated with the active phase of UC. METHODS: The GEO database was searched for UC-related datasets that included healthy controls who met the screening criteria. Differential analysis was conducted to obtain differentially expressed genes (DEGs). Autophagy-related targets were collected and intersected with the DEGs to identiy differentially expressed autophagy-related genes (DEARGs) associated with active UC. DEARGs were then subjected to KEGG, GO, and DisGeNET disease enrichment analyses using R software. Differential analysis of immune infiltrating cells was performed using the CiberSort algorithm. The least absolute shrinkage and selection operator algorithm and protein-protein interaction network were used to narrow down the DEARGs, and the top five targets in the Dgree ranking were designated as core targets. RESULTS: A total of 4822 DEGs were obtained, of which 58 were classified as DEARGs. SERPINA1, BAG3, HSPA5, CASP1, and CX3CL1 were identified as core targets. GO enrichment analysis revealed that DEARGs were primarily enriched in processes related to autophagy regulation and macroautophagy. KEGG enrichment analysis showed that DEARGs were predominantly associated with NOD-like receptor signaling and other signaling pathways. Disease enrichment analysis indicated that DEARGs were significantly linked to diseases such as malignant glioma and middle cerebral artery occlusion. Immune infiltration analysis demonstrated a higher presence of immune cells like activated memory CD4 T cells and follicular helper T cells in active UC patients than in healthy controls. CONCLUSION: Autophagy is closely related to the active phase of UC and the potential targets obtained from the analysis in this study may provide new insight into the treatment of active UC patients.

[Effects of Straw Biochar on Carbon Footprint of Maize Farmland Ecosystem Under Mulched Drip Irrigation in Hetao Irrigation District].

To explore the effect of biochar on greenhouse gas emissions and the carbon footprint of a corn farmland ecosystem under drip irrigation with film in an arid region, biochar treatments with different application rates[0 (CK), 15 (C15), 30 (C30), and 45 t·hm-2 (C45)] were established. The seasonal changes in soil greenhouse gases (CO2, N2O, and CH4) and their comprehensive warming potential in the maize farmland ecosystem were monitored for two consecutive years after a one-time application of biochar. The carbon emissions caused by agricultural production activities and their carbon footprint were estimated using the life cycle assessment method. Compared with that in CK, the cumulative CO2 emissions in the crop growing season decreased by 17.6%-24.7%, the cumulative N2O emissions decreased by 71.1%-110.4%, and the global warming potential decreased by 19.5%-25.9%. In the second year of the crop growing season after biochar application, the cumulative CO2 emissions were reduced by 19.2%-40.6%, the cumulative N2O emissions were reduced by 38.7-46.7%, and the comprehensive warming potential was reduced by 19.7%-40.5%. For two consecutive years, the treatment of C15 and C30 increased the cumulative absorption of CH4 to different degrees, whereas the treatment of C45 significantly decreased the cumulative absorption of CH4. C15 and C45 were the treatments with the least carbon footprint per unit yield in the current and the succeeding year of biochar application, and their carbon footprint per unit yield was 10.1% and 26.2% lower than that of CK, respectively. Soil greenhouse gas emissions showed the most contribution to the carbon footprint of the maize farmland ecosystem (38.1%-59.2%), followed by nitrogen fertilizer production (19.8%-33.4%), electric energy production (6.7%-8.8%), and plastic film mulching (4.4%-7.4%). Biochar contributed 5.7%-13.8% to the ecosystem's carbon footprint. The application of 30 t·hm-2 biochar had a better effect on carbon reduction, carbon fixation, and yield increase in the farmland ecosystem. Improving the biochar production process and transportation route, increasing nitrogen use efficiency, and developing water-saving and energy-saving irrigation technology are important ways to reduce the carbon footprint of farmland ecosystems in arid regions.

Effects of sowing patterns on nitrogen utilization and yield formation of winter wheat in the western Huang-Huai-Hai region.

To examine the differences of three improved sowing methods in winter wheat yield and nitrogen efficiency and reveal the characteristics responsible for such differences, we conducted field experiments in the Jinnan area of the western Huang-Huai-Hai wheat region for three consecutive seasons from 2016 to 2019. The three improved sowing methods were wide space sowing (WSS), furrow sowing in moisture soil (FS), and three-dimensional uniform sowing (TDUS), with conventional drilling sowing (CDS) as the control. The results showed that meteorological factors such as accumulated temperature, solar radiation, and precipitation in the growing seasons from 2016 to 2019 showed great intra- and inter-annual variations. Compared with CDS, the improved sowing methods (WSS, FS, and TDUS) enhanced spike number per unit area and increased grain yield in three growing seasons by 18.3%-55.5%, 8.6%-22.2%, and 10.9%-39.5%, respectively. The three methods increased nitrogen uptake efficiency (NEup) by 5.8%-57.1%, pre-flowering nitrogen transfer ratio (Np/Nt) by 3.0%-15.3%, and nitrogen efficiency by 7.9%-35.7%, respectively. We developed a structural equation model (SEM) by integrating meteorological factors and experimental variables. The results showed that the three improved sowing methods could reduce the effects of extreme low temperature on wheat plant population, increase NEup and Np/Nt, and provide sufficient nitrogen supply to the grains of high-spike number wheat population for high yield and high nitrogen efficiency. In summary, our results demonstrated that WSS, FS, and TDUS all improved NEup and Np/Nt in the 2016-2017 season when meteorological conditions were favorable for wheat growth, and enhanced yield components with high SN, leading to high yield and high nitrogen efficiency. In contrast, in both 2017-2018 and 2018-2019 seasons with extremely low temperature and uneven distribution of meteorological conditions, WSS had a higher number of tillers at the jointing stage and enhanced pre-flowering nitrogen uptake and translocation, whereas TDUS had a relatively stable nitrogen uptake rate, leading to a stable grain yield.

Opportunities and challenges for ribosome-inactivating proteins in traditional Chinese medicine plants.

Ribosome-inactivating proteins (RIPs) are a class of cytotoxic rRNA N-glycosylase, which widely exist in higher plants in different taxonomy, including many traditional Chinese medicinal materials and vegetables and fruits. In this paper, the traditional Chinese medicinal plants containing RIPs protein were sorted out, and their pharmacological effects and clinical applications were analyzed. Since many RIPs in traditional Chinese medicine plants exhibit antiviral and antitumor activities and show great clinical application potential, people's interest in these proteins is on the rise. This paper summarizes the possible mechanism of RIPs's anti-virus and anti-tumor effects, and discusses its potential problems and risks, laying a foundation for subsequent research on how to exert its anti-virus and anti-tumor effects.

Immunological effects of recombinant Lactobacillus casei expressing pilin MshB fused with cholera toxin B subunit adjuvant as an oral vaccine against Aeromonas veronii infection in crucian carp.

Aeromonas veronii is a zoonotic agent capable of infecting fish and mammals, including humans, posing a serious threat to the development of aquaculture and public health safety. Currently, few effective vaccines are available through convenient routes against A. veronii infection. Herein, we developed vaccine candidates by inserting MSH type VI pili B (MshB) from A. veronii as an antigen and cholera toxin B subunit (CTB) as a molecular adjuvant into Lactobacillus casei and evaluated their immunological effect as vaccines in a crucian carp (Carassius auratus) model. The results suggested that recombinant L. casei Lc-pPG-MshB and Lc-pPG-MshB-CTB can be stably inherited for more than 50 generations. Oral administration of recombinant L. casei vaccine candidates stimulated the production of high levels of serum-specific immunoglobulin M (IgM) and increased the activity of acid phosphatase (ACP), alkaline phosphatase (AKP) superoxide dismutase (SOD), lysozyme (LZM), complement 3 (C3) and C4 in crucian carp compared to the control group (Lc-pPG612 group and PBS group) without significant changes. Moreover, the expression levels of interleukin-10 (IL-10), interleukin-1ß (IL-1ß), tumour necrosis factor-α (TNF-α) and transforming growth factor-ß (TGF-ß) genes in the gills, liver, spleen, kidney and gut of crucian carp orally immunized with recombinant L. casei were significantly upregulated compared to the control groups, indicating that recombinant L. casei induced a significant cellular immune response. In addition, viable recombinant L. casei can be detected and stably colonized in the intestine tract of crucian carp. Particularly, crucian carp immunized orally with Lc-pPG-MshB and Lc-pPG-MshB-CTB exhibited higher survival rates (48% for Lc-pPG-MshB and 60% for Lc-pPG-MshB-CTB) and significantly reduced loads of A. veronii in the major immune organs after A. veronii challenge. Our findings indicated that both recombinant L. casei strains provide favorable immune protection, with Lc-pPG-MshB-CTB in particular being more effective and promising as an ideal candidate for oral vaccination.

Two new dinor-eudesmane sesquiterpenoids from the roots of Chloranthus multistachys.

Two new dinor-eudesmane sesquiterpenoids, named multistalin A (1), and multistalin B (2), together with three sesquiterpene glycosides (3-5), and a norlabdane-type diterpene (6) were isolated from the root extract of Chloranthus multistachys Pei. Their structures were elucidated on the basis of spectroscopic analysis including 1D, 2D NMR techniques and HR-ESI-MS. In addition, the cytotoxicity activities of the isolated compounds against selected cancer cells (Hela and A-549) were evaluated by MTT assay.

[Changes and Potential Sources of Atmospheric Black Carbon Concentration in Shanghai over the Past 40 Years Based on MERRA-2 Reanalysis Data].

As an important component of atmospheric aerosols, black carbon (BC) has a great influence on the regional and global radiation balance, climate, and human health due to its small particle size, large specific surface area, and radiative forcing potential. Here, the spatio-temporal characteristics of atmospheric BC were investigated based on modern-era retrospective analysis for research and applications version 2 (MERRA-2) reanalysis data and ground observation data during 1980-2019 in Shanghai, a highly urbanized city in mainland China. The influences of local emissions and regional transmission on regional-scale BC concentrations were examined using the M-K trend test, backward trajectory analysis, and the potential source contribution function (PSCF). The results showed that:① MERRA-2 BC and ground observation datasets showed good consistency (R∈[0.68, 0.72]), indicating that MERRA-2 reanalysis data can be used to reveal long-term changes in ground-level atmospheric BC concentrations; ② Atmospheric BC concentrations in Shanghai over the past 40 years can be divided into three stages:a "low value" stage of slow growth[1980-1986, (1.75±0.17) µg·m-3], a relatively stable "median value" stage[1987-1999, (2.18 ±0.07) µg·m-3], and a fluctuating "high value" stage[2000-2019, (3.07±0.31) µg·m-3]. Seasonally, Shanghai's BC concentrations generally show a "U" pattern with low concentrations in summer and high concentrations in winter. As a result of black carbon emissions from marine diesel engines and other engines used for water transportation, a small peak also occurs in July; ③ The diagnostic quality ratio of air pollutants and the bivariate correlation analysis[R(BC-NO2)>R(BC-CO)>R(BC-SO2)] indicated that traffic emissions were the main sources of atmospheric BC in Shanghai, especially by heavy diesel vehicles; ④ The backward trajectory and PSCF analyses found that the air mass of Shanghai in summer was dominated by a clean sea breeze, accounting for 77.18%. In contrast, during the other seasons, more than 50% of the air mass came from the north. The potential source regions of atmospheric BC in Shanghai are mainly distributed in eastern China, expanding outwards and centering on the Yangtze River Delta, and the expansion direction is consistent with the directions of the backward trajectories.

Fluorescent polydopamine nanoparticles as a nanosensor for the sequential detection of mercury ions and l-ascorbic acid based on a coordination effect and redox reaction.

Herein, a novel fluorescence nanosensor using intrinsic fluorescent polydopamine nanoparticles (PDA NPs) as an effective signal reporter has been constructed for the simple, rapid and sequential detection of mercury ions (Hg2+) and l-ascorbic acid (AA) based on a coordination effect and redox reaction. The fluorescence of the PDA NPs could be specifically quenched by Hg2+ through intense coordination effects between the Hg2+ and the groups (catechol, amine, ketone and imine) on the surface of the PDA NPs. However, when AA and Hg2+ coexisted in solution, the fluorescence of the PDA NPs pronouncedly recovered via the redox reaction of Hg2+, with it being reduced to Hg0 by AA. The fluorescence quenching mechanism of Hg2+ towards the PDA NPs and the redox reaction between Hg2+ and AA were also fully investigated. The nanosensor exhibited high sensitivity and desirable selectivity for Hg2+ and AA detection. Moreover, the strategy was successfully explored in real samples (tap water, lake water and human serum samples) with satisfactory recoveries. The developed nanosensor provides new sights and good inspiration for Hg2+ and AA detection under real conditions.

Effects of probiotics on nonalcoholic fatty liver disease: a systematic review and meta-analysis.

BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) has become prevalent in recent decades, especially in developed countries, and approaches for the prevention and treatment of NAFLD are not clear. The aim of this research was to analyze and summarize randomized controlled trials that investigated the effects of probiotics on NAFLD. METHODS: Seven databases (PubMed, Embase, the Web of Science, the Cochrane Library, China National Knowledge Infrastructure, Wan Fang Data, and VIP Database) were searched. Then, eligible studies were identified. Finally, proper data extraction, synthesis and analysis were performed by trained researchers. RESULTS: Anthropometric parameters: with use of probiotics weight was reduced by 2.31 kg, and body mass index (BMI) was reduced by 1.08 kg/m2. Liver function: probiotic treatment reduced the alanine aminotransferase level by 7.22 U/l, the aspartate aminotransferase level by 7.22 U/l, the alkaline phosphatase level by 25.87 U/l, and the glutamyl transpeptidase level by -5.76 U/l. Lipid profiles: total cholesterol, low-density lipoprotein cholesterol, and triglycerides were significantly decreased after probiotic treatment. Their overall effects (shown as standard mean difference) were -0.73, -0.54, and -0.36, respectively. Plasma glucose: probiotics reduced the plasma glucose level by 4.45 mg/dl and the insulin level by 0.63. Cytokines: probiotic treatment decreased tumor necrosis factor alpha by 0.62 and leptin by 1.14. Degree of liver fat infiltration (DFI): the related risk of probiotics for restoring DFI was 2.47 (95% confidence interval, 1.61-3.81, p < 0.001). CONCLUSION: Probiotic treatment or supplementation is a promising therapeutic method for NAFLD.

Effect of haptoglobin on the treatment of chronic hepatitis B with interferon.

There are two main types of drugs that are used to treat chronic hepatitis B (CHB), including interferon (IFN) and nucleotide analogues. IFN inhibits the virus through direct antiviral activity and via immune regulation, and it has been widely applied for the treatment of CHB and other infections. However, the efficiency of IFN therapy is not entirely satisfactory. The aim of the present study was to investigate the factors affecting IFN therapy. The plasma of patients with CHB treated with IFN was collected and divided into the virological response group and non-virological response group according to their virological response. Serum proteins of virologically responsive patients were compared before and after IFN therapy using isobaric tags for relative and absolute quantitation technology. ELISA was used to validate these results in the same sample. In in vitro cell experiments, HepG2.2.15 cells were transfected with haptoglobin (Hp)-targeting small interfering RNA to inhibit expression of the Hp protein, and reverse transcription-quantitative polymerase chain reaction and western blotting were utilized to detect hepatitis B virus (HBV)-DNA, IFN and downstream molecular changes in the cell supernatants. The Hp protein levels were demonstrated to be significantly lower following 48 weeks of IFN therapy, and the levels of Hp in patients in the virological response group were significantly lower than those in the non-virological response group. In in vitro cell experiments, following inhibition of Hp protein expression, significantly decreased levels of HBV-DNA, and elevated levels of IFN and its downstream molecules were observed. These findings suggest that Hp may be able to predict the efficacy of IFN therapy, and it may inhibit HBV clearance. There is an association between Hp and IFN, which requires further clinical and laboratory studies to explore.

Sorcin induces gastric cancer cell migration and invasion contributing to STAT3 activation.

Gastric cancer (GC) is a globally occurring malignancy that is characterized by a high mortality rate due to a high tendency to metastasize and poor prognoses. Sorcin, as known as SRI, a soluble resistance-related calcium-binding protein, plays a significant role in multidrug resistance. Sorcin is related to the migration and invasion of cancer cells. However, the mechanism remains unclear. Here, we used immunohistochemistry to confirm that the expression of sorcin in cancer tissues is higher than that in the adjacent normal tissues. The wound healing and transwell results indicate that sorcin can induce migration and invasion of GC cells. To explore the role of sorcin in GC metastasis, isobaric tags for relative and absolutely quantitation (iTRAQ) were used to examine cells with and without sorcin knockdown to identify the differentially expressed proteins (DEPs). The results were evaluated via RT-PCR and western blot to confirm the ITRAQ data. Inhibition of sorcin expression can down- regulate the expression of CTSZ, MMP2, MMP9 and p-STAT3 followed by suppression of tumor growth and metastasis. Together, we concluded that sorcin has a oncogenic activity via inducing tumor growth and metastasis, leading to development of therapeutic treatments for GC.

[Research Progress on the Genomics of Taenia solium and Candidate Vaccines for Cysticercosis].

Research on cysticercosis has been stimulated by the rapid development of genomics, proteomics and bioinformatics and the emergence of molecular biological and immunological technologies. In this review, we attempt to discuss the research development on genomics of Taenia solium and candidate vaccines and antigens for cysticercosis. This will provide a new perspective for studying genomics of Taenia solium and for cysticercosis prevention and treatment, and provide a wealth of informative data for the development of novel and highly efficient vaccines against cysticercosis or diagnostic antigen molecules for cysticercosis.

Keratin 8 is involved in hepatitis B virus replication.

Hepatitis B virus (HBV) infection can result in fatal liver diseases, including cirrhosis or liver failure, and its replication and pathogenesis depend on the critical interplay between viral and host factors. This study investigated HBV replication-related host proteins and the effect of candidate proteins on HBV replication. Isobaric tags for relative and absolute quantitation (iTRAQ) were used to measure HBV replication-related proteins in HepG2 cells and HepG2.2.15 cells. KRT8 was up-regulated in HepG2.2.15 cells but not in HepG2 cells, and KRT8 was overexpressed in an HBV-infected patient's liver tissue. This result suggested that KRT8 is involved in HBV replication. To further clarify the relationship between KRT8 and HBV replication, KRT8 gene expression was inhibited by siRNA. The silencing of KRT8 mildly suppressed HBV replication. Moreover, overexpressed KRT8 significantly increased HBV replication, and the inhibition of HBV DNA did not suppress KRT8 expression. Thus, the host protein KRT8 is involved in the replication of HBV DNA, and it dramatically enhances HBV replication.

HSPB1 is an intracellular antiviral factor against hepatitis B virus.

Hepatitis B virus (HBV) is the most common of the hepatitis viruses that cause chronic liver infections in humans and it is considered a major global health problem. However, the mechanisms of HBV replication are complex and not yet fully understood. In this study, the HBV DNA-transfected HepG2.2.15 cell line and its parental HepG2 cell line were analyzed by isobaric tags for relative and absolute quantitation (iTRAQ)-coupled two-dimensional liquid chromatography tandem mass-spectrophotometry (2D LC-MS/MS), a successfully exploited high-throughput proteomic technology. In total, 2,028 unique proteins were identified and 170 proteins were differentially expressed in HepG2.2.15 cells as compared with that in HepG2. Several differentially expressed proteins were further validated by Western blot and real-time quantitative reverse transcription-PCR. Furthermore, the association of HBV replication with heat shock protein B1, one of the highly expressed proteins in HepG2.2.15 cells, was verified. HSPB1 functions as a anti-viral protein during HBV infection by specifically inducing type interferon and some downstream antiviral effectors. This study is the first to report the application of iTRAQ technology to analyze the underlying mechanisms of HBV replication. Many of the differentially expressed proteins identified have not been linked to HBV replication before, and may provide valuable novel insights into HBV replication.

Quantitative proteome analysis of ovarian cancer tissues using a iTRAQ approach.

Quantitative proteomics can be used as a screening tool for identification of differentially expressed proteins as potential biomarkers for cancers. Here, we comparatively analyzed the proteome profiles of ovarian cancer tissues and normal ovarian epithelial tissues. Using the high-throughput proteomic technology of isobaric tags for relative and absolute quantitation (iTRAQ)-coupled with two-dimensional-liquid chromatography-tandem mass spectrometry, 1,259 unique proteins were identified. Of those, 205 were potentially differentially expressed between ovarian cancer and normal ovarian tissues. Several of the potentially differentially expressed proteins were validated by Western blotting and real-time quantitative RT-PCR analyses. Furthermore, up-regulation of KRT8, PPA1, IDH2, and S100A11 were validated in ovarian tissue microarrays by immunohistochemistry. Silencing of S100A11 expression suppressed the migration and invasion properties of ovarian cancer cells in vitro. Our study represents the successful application of iTRAQ technology to an investigation of ovarian cancer. Many of the potentially differentially expressed proteins identified had not been linked to ovarian cancer before, and provide valuable novel insights into the underlying mechanisms of carcinogenesis in human ovarian cancer.

Aplasia ras homolog member I is downregulated in gastric cancer and silencing its expression promotes cell growth in vitro.

BACKGROUND AND AIM: Aplasia ras homolog member I (ARHI) is a maternally imprinted tumor suppressor gene. ARHI protein is widely expressed in many types of human tissues; however, its expression is frequently reduced or absent in various tumors and plays a tumor suppressor role for in vitro study. In this study, we investigated the expression level of ARHI in gastric cancer in order to investigate the function of ARHI and signaling pathways that might be linked during gastric cancer development. METHODS: ARHI mRNA and protein expression levels were analyzed in primary gastric cancer tissues, adjacent noncancerous gastric tissues and gastric cancer cell lines using semi-quantitative polymerase chain reaction, western blotting and immunohistochemistry, respectively. RESULTS: Our results showed that both mRNA and protein expression levels of the ARHI gene were significantly downregulated (P < 0.05) in gastric cancer tissues and cell lines compared to the corresponding normal control groups. The protein expression level of ARHI was not associated with age, gender, location of tumor, tumor size or metastasis in patients with gastric cancer. However, a significant correlation between the level of ARHI protein expression and the degree of tumor differentiation and Tumor-Node-Metastasis stage was observed (P < 0.05). Furthermore, results of the methyl thiazolyl tetrazolium and Transwell assays and flow cytometric analysis showed increased cell proliferation, migration and anti-apoptotic capacities in the well-differentiated gastric cancer MKN-28 cell line, which has stably silenced ARHI protein expression. CONCLUSION: Our data indicate that ARHI expression is downregulated in human gastric cancer and it may be a novel tumor suppressive target for gastric cancer therapy.

Proteomic investigation of 5-fluorouracil resistance in a human hepatocellular carcinoma cell line.

Multi-drug resistance (MDR) is a major obstacle towards a successful treatment of hepatocellular carcinoma (HCC). The mechanisms of MDR are intricate and have not been fully understood. Therefore, we employed a cell-line model consisting of the 5-fluorouracil (5-FU) resistant BEL7402/5-FU cell line and its parental BEL7402 cell line. Using relative and absolute quantification (iTRAQ)-coupled 2D LC-MS/MS, a successfully exploited high-throughput proteomic technology, in total, 660 unique proteins were identified and 52 proteins showed to be differentially expressed in BEL7402/5-FU compared with BEL7402. Several differentially expressed proteins were further validated by Western blot and real-time quantitative RT-PCR analysis. Furthermore, the association of MDR with ANXA3, one of the highly expressed proteins in BEL7402/5-FU, was verified. Our study represents the first successful application of iTRAQ technology for MDR mechanisms analysis in HCC. Many of the differentially expressed proteins identified had not been linked to MDR in HCC before, which provide valuable information for further understanding of MDR.

[Expression and clinical significance of BRCA1 in human esophageal squamous cell carcinoma].

OBJECTIVE: To investigate the expression of BRCA1 in esophageal squamous cell carcinoma (ESCC) tissues and evaluate its correlation with clinicopathological features as well as the prognosis of ESCC patients. METHODS: The expression of BRCA1 was detected by immunohistochemistry (IHC) in 201 specimens of T3 stage ESCC tissues and corresponding adjacent normal tissues using tissue microarray. The correlation between BRCA1 expression and clinicopathological features of ESCC was determined by chi-square analysis. The cumulative survival rate was analyzed by Kaplan-Meier method. RESULTS: The positive rate of BRCA1 expression in ESCC tissues was significantly higher than that in adjacent normal tissues [88.6% (178/201) vs. 36.8% (74/201), P < 0.001]. There was a significant correlation between the expression of BRCA1 and lymph node metastasis. In the tumors with positive lymph nodes, strong positive expression of BRCA1 was found in 45.0% (49/109), while only 19.6% (18/92) in tumors without lymph node metastasis, showing a significant difference (P < 0.001). A close relationship was also found between the expression of BRCA1 and gross typing of tumors (P < 0.05). The expression of BRCA1 was not significantly correlated with gender, age, tumor location, differentiation, and tumor thrombus (P > 0.05). The results of Kaplan-Meier analysis indicated that ESCC patients with a higher positive rate of BRCA1 expression have a poorer prognosis (P < 0.05). CONCLUSIONS: The expression of BRCA1 is related to the occurrence and development of esophageal carcinoma. BRCA1 protein may serve as a new potential biomarker in estimating the biological behavior of ESCC.

Preparation of novel anti-ski monoclonal antibodies.

Ski is an avian sarcoma virus oncogene homolog best known for inhibiting TGF beta signaling through its association with the SMAD proteins. Anti-Ski antibodies (MAbs) of high titer were prepared by immunizing BALB/c mice with multifocal intradermal injections and fusing high titer antibody producing spleen cells with myeloma cells of SP2/0 origin. Three MAbs were selected for further characterization as classes and subclasses. Antibodies were produced by these three clones with high affinities ranging from 10(9) to 10(11)/m. These clones were found to be of the immunoglobulin IgG1 and IgG2b subclass with kappa light chain. They could recognize Ski as determined by Western blot analysis. The produced MAbs will be a useful tool for further investigation of Ski functions in organisms.

Comparative proteome analysis of 3T3-L1 adipocyte differentiation using iTRAQ-coupled 2D LC-MS/MS.

Adipose tissue is critical in obesity and type II diabetes. Blocking of adipocyte differentiation is one of the anti-obesity strategies targeting on strong rise in fat storage and secretion of adipokine(s). However, the molecular basis of adipocyte differentiation and its regulation remains obscure. Therefore, we exposed 3T3-L1 cell line to appropriate hormonal inducers as adipocyte differentiation model. Using iTRAQ-coupled 2D LC-MS/MS, a successfully exploited high-throughput proteomic technology, we nearly quantitated 1,000 protein species and found 106 significantly altered proteins during adipocyte differentiation. The great majority of differentially expressed proteins were related to metabolism enzymes, structural molecules, and proteins involved in signal transduction. In addition to previously reported differentially expressed molecules, more than 20 altered proteins previously unknown to be involved with adipogenic process were firstly revealed (e.g., HEXB, DPP7, PTTG1IP, PRDX5, EPDR1, SPNB2, STEAP3, TPP1, etc.). The partially differential proteins were verified by Western blot and/or real-time PCR analysis. Furthermore, the association of PCX and VDAC2, two altered proteins, with adipocyte conversion was analyzed using siRNA method, and the results showed that they could contribute considerably to adipogenesis. In conclusion, our data provide valuable information for further understanding of adipogenesis.