CTLA4 expression profiles and their association with clinical outcomes of breast cancer: a systemic review.

Purpose: The cytotoxic T-lymphocyte-associated protein 4 (CTLA4) is involved in the progression of various cancers, but its biological roles in breast cancer (BRCA) remain unclear. Therefore, we performed a systematic multiomic analysis to expound on the prognostic value and underlying mechanism of CTLA4 in BRCA. Methods: We assessed the effect of CTLA4 expression on BRCA using a variety of bioinformatics platforms, including Oncomine, GEPIA, UALCAN, PrognoScan database, Kaplan-Meier plotter, and R2: Kaplan-Meier scanner. Results: CTLA4 was highly expressed in BRCA tumor tissue compared to normal tissue (P < 0.01). The CTLA4 messenger RNA levels in BRCA based on BRCA subtypes of Luminal, human epidermal growth factor receptor 2, and triple-negative BRCA were considerably higher than in normal tissues (P < 0.001). However, the overexpression of CTLA4 was associated with a better prognosis in BRCA (P < 0.001) and was correlated with clinicopathological characteristics including age, T stage, estrogen receptors, progesterone receptors, and prediction analysis of microarray 50 (P < 0.01). The infiltration of multiple immune cells was associated with increased CTLA4 expression in BRCA (P < 0.001). CTLA4 was highly enriched in antigen binding, immunoglobulin complexes, lymphocyte-mediated immunity, and cytokine-cytokine receptor interaction. Conclusion: This study provides suggestive evidence of the prognostic role of CTLA4 in BRCA, which may be a therapeutic target for BRCA. Furthermore, CTLA4 may influence BRCA prognosis through antigen binding, immunoglobulin complexes, lymphocyte-mediated immunity, and cytokine-cytokine receptor interaction. These findings help us understand how CTLA4 plays a role in BRCA and set the stage for more research.

Efficacy of electroacupuncture in preventing nausea and vomiting after thyroidectomy: A prospective randomized controlled trial.

OBJECTIVE: Postoperative nausea and vomiting are regarded as a serious concern after thyroidectomy. Electroacupuncture shows the potential to reduce general anesthesia-related side effects. The aim of this study was to evaluate the efficacy of electroacupuncture in preventing postoperative nausea and vomiting that commonly occurs in patients after thyroidectomy. METHODS: This study was a prospective randomized controlled trial with a two-arm, patient blind structure. Sixty-four participants were randomly assigned to the acupuncture (n = 35) or control (n = 29) group. Patients in the acupuncture group received electroacupuncture and intradermal press needles. The primary endpoint was the incidence and severity of postoperative nausea and vomiting, and secondary endpoints were the length of hospital stay, pain severity, and postoperative in-hospital morbidity. RESULTS: The total incidence of postoperative nausea and vomiting was 40.6% (26/64). There was no difference in the incidence between the control (10/29, 34.5%) and acupuncture (16/35, 45.7%) groups (p = 0.362). The severity of postoperative nausea and vomiting was not different between the groups (p = 0.842). Length of hospitalization and postoperative complications were not different between the groups. CONCLUSION: In this randomized controlled trial, electroacupuncture treatment after thyroidectomy is safe and comparable to conventional anti-emetic therapy. TRIAL REGISTRATION: Clinical Research Information Service, KCT0001782. Registered on 26 January 2016.

Vitamin D receptor (VDR) mRNA overexpression is associated with poor prognosis in breast carcinoma.

Purpose: The prognostic value of vitamin D receptor gene (VDR) expression in breast cancer development is unclear. Here, we aimed to investigate whether VDR expression can be used as a prognostic indicator of breast cancer. Methods: We used various public bioinformatics platforms: Oncomine, GEPIA, UALCAN, Kaplan-Meier plotter, UCSC XENA, bc-GenExMiner, WebGestalt, and STRING database. Results: We found that VDR was upregulated in breast cancer in comparison to normal tissues. Overexpression of VDR was significantly associated with worse overall survival in breast cancer. The expression of VDR was related to age, TNM stages, estrogen receptor status, progesterone receptor status, human epidermal growth factor receptor 2 status, basal-like (PAM 50) status, triple-negative breast cancer (TNBC) status, and basal-like (PAM 50) & TNBC status (P < 0.05). Increased VDR expression in breast cancer was significantly associated with older age. The 5 hub genes for VDR were NCOA1, EP300, CREBBP, and RXRA. Conclusion: Our investigation offers hints about the prognostic role of VDR in breast cancer. The findings suggest that VDR expression might be used as a marker to determine a breast cancer patient's prognosis. Nevertheless, further validation is warranted.

Identification and Characterization of a Bacteriocin from the Newly Isolated Bacillus subtilis HD15 with Inhibitory Effects against Bacillus cereus.

Natural antimicrobial substances are needed as alternatives to synthetic antimicrobials to protect against foodborne pathogens. In this study, a bacteriocin-producing bacterium, Bacillus subtilis HD15, was isolated from doenjang, a traditional Korean fermented soybean paste. We sequenced the complete genome of B. subtilis HD15. This genome size was 4,173,431 bp with a G + C content of of 43.58%, 4,305 genes, and 4,222 protein-coding genes with predicted functions, including a subtilosin A gene cluster. The bacteriocin was purified by ammonium sulfate precipitation, Diethylaminoethanol-Sepharose chromatography, and Sephacryl gel filtration, with 12.4-fold purification and 26.2% yield, respectively. The purified protein had a molecular weight of 3.6 kDa. The N-terminal amino acid sequence showed the highest similarity to Bacillus subtilis 168 subtilosin A (78%) but only 68% similarity to B. tequilensis subtilosin proteins, indicating that the antimicrobial substance isolated from B. subtilis HD15 is a novel bacteriocin related to subtilosin A. The purified protein from B. subtilis HD15 exhibited high antimicrobial activity against Listeria monocytogenes and Bacillus cereus. It showed stable activity in the range 0-70°C and pH 2-10 and was completely inhibited by protease, proteinase K, and pronase E treatment, suggesting that it is a proteinaceous substance. These findings support the potential industrial applications of the novel bacteriocin purified from B. subtilis HD15.

BRCA1/2 Serves as a Biomarker for Poor Prognosis in Breast Carcinoma.

BRCA1/2 are breast cancer susceptibility genes that are involved in DNA repair and transcriptional control. They are dysregulated in breast cancer, making them attractive therapeutic targets. Here, we performed a systematic multiomics analysis to expound BRCA1/2 functions as prognostic biomarkers in breast cancer. First, using different web-based bioinformatics platforms (Oncomine, TIMER 2.0, UALCAN, and cBioportal), the expression of BRCA1/2 was assessed. Then, the R package was used to analyze the diagnostic value of BRCA1/2 in patients. Next, we determined the relationship between BRCA1/2 mRNA expression and prognosis in patients (PrognoScan Database, R2: Kaplan Meier Scanner and Kaplan−Meier Plotter). Subsequently, the association of BRCA1/2 with mutation frequency alteration and copy number alterations in breast cancer was investigated using the cBioportal platform. After that, we identified known and predicted structural genes and proteins essential for BRCA1/2 functions using GeneMania and STRING db. Finally, GO and KEGG pathway enrichment analyses were performed to elucidate the potential biological functions of the co-expression genes of BRCA1/2. The BRCA1/2 mRNA level in breast cancer tissues was considerably higher than in normal tissues, with AUCs of 0.766 and 0.829, respectively. Overexpression of BRCA1/2 was significantly related to the worse overall survival (p < 0.001) and was correlated to clinicopathological characteristics including lymph nodes, estrogen receptors, and progesterone receptors (p < 0.01). The alteration frequencies of both the gens have been checked, and the results show that BRCA1 and BRCA2 show different alteration frequencies. Their mutation sites differ from each other. GO and KEGG showed that BRCA1/2 was mainly enriched in catalytic activity, acting on DNA, chromosomal region, organelle fission, cell cycle, etc. The 20 most frequently changed genes were closely related to BRCA1/2, including PALB2 and RAD51 relatively. Our study provides suggestive evidence of the prognostic role of BRCA1/2 in breast cancer and the therapeutic target for breast cancer. Furthermore, BRCA1/2 may influence BRCA prognosis through catalytic activity, acting on DNA, chromosomal regions, organelle fission, and the cell cycle. Nevertheless, further validation is warranted.

Transoral Endoscopic Thyroidectomy Vestibular Approach (TOETVA): Surgical Outcomes and Learning Curve.

The transoral endoscopic thyroidectomy vestibular approach (TOETVA) has excellent cosmetic effects and its popularity is increasing worldwide. We present our experience with TOETVA and its short-term outcomes. This study included 110 consecutive patients who underwent TOETVA at a single institution between July 2016 and June 2020. We analyzed clinicopathologic data, short-term postoperative outcomes, and learning curve using cumulative summation (CUSUM) analysis. Of the 110 patients who underwent TOETVA, 101 had malignant disease and 100 (90.9%) underwent lobectomy. The mean age was 39.7 ± 9.7 years, and the mean tumor size was 1.0 ± 0.7 cm (range, 0.3-3.6 cm). Operation time was 168.0 ± 63.4 min for total thyroidectomy, 111.0 ± 27.7 min for lobectomy, and 73.7 ± 18.1 min for isthmusectomy. Five patients (4.5%) experienced transient vocal cord palsy (VCP) and one (0.9%) had permanent VCP. The swallowing impairment index-6 score was 2.18 ± 3.21 at postoperative three months, and 0.97 ± 1.72 at postoperative six months. The learning curve for lobectomy was 58 cases in CUSUM analysis. TOETVA is a safe and feasible approach with an acceptable operation time and a low complication rate. This approach is a surgical option for patients who desire excellent cosmesis.

Selective inhibition of V600E-mutant BRAF gene induces apoptosis in thyroid carcinoma cell lines.

PURPOSE: Papillary thyroid cancer (PTC) has a high incidence of BRAFV600E mutation. The purpose of this study was to evaluate the potential relationship between thyroiditis and BRAFV600E mutation status in patients with PTC. We investigated how a selective inhibitor of BRAFV600E PLX4032 affects the proliferation and inflammatory cytokine levels of thyroid cancer. METHODS: Two thyroid cancer cell lines TPC1 and 8505C were treated with PLX4032, an analysis was done on cell growth, cell cycle, the degree of apoptosis, and levels of inflammatory cytokines. To identify the functional links of BRAF, we used the STRING database. RESULTS: Docking results illustrated PLX4032 blocked the kinase activity by exclusively binding on the serine/threonine kinase domain. STRING results indicated BRAF is functionally linked to mitogen-activated protein kinase. Both cell lines showed a dose-dependent reduction in growth rate but had a different half maximal inhibitory concentration value for PLX4032. The reaction to PLX4032 was more sensitive in the 8505C cells than in the TPC1 cells. PLX4032 induced a G2/M phase arrest in the TPC1 cells and G0/G1 in the 8505C cells. PLX4032 induced apoptosis only in the 8505C cells. With PLX4032, the TPC1 cells showed decreased levels of vascular endothelial growth factor, granulocyte-macrophage colony-stimulating factor, chemokine (C-C motif) ligand 2/monocyte chemoattractant protein 1, whereas the 8505C cells showed significantly decreased levels of IL-8, serpin E1/plasminogen activator inhibitor-1, and matrix metalloproteinase (MMP)-3. CONCLUSION: PLX4032 was cytotoxic in both TPC1 and 8505C cells and induced apoptosis. In the 8505C cells, inflammatory cytokines such as IL-8 and MMP-3 were down-regulated. These findings suggest the possibility that the BRAFV600E mutation needs to target inflammatory signaling pathways in the treatment of thyroid cancer.

Shear-wave elastography in thyroid ultrasound: Can be a predictor of extrathyroidal extension and cervical lymph node metastasis in papillary thyroid carcinoma?

ABSTRACT: This study aimed to investigate whether extrathyroidal extension (ETE) and cervical lymph node metastasis (LNM) can be predicted using elasticity parameters of shear-wave elastography (SWE) combined with B-mode ultrasound (US) of papillary thyroid carcinomas (PTCs).We retrospectively reviewed 111 patients who underwent preoperative SWE evaluation among PTC patients from July 1, 2016 to June 20, 2018. Patients were divided into 2 groups based on the presence or absence of ETE based on pathology reports. Univariate and multivariate analyses of clinical and radiologic features including B-mode US features, US patterns, and SWE parameters were performed. These analyses were repeated in LNM positive and negative groups. The diagnostic performance of SWE parameters were also evaluated.Of the 111 patients, 33 had ETE, 78 did not have ETE, 44 had LNM, and 67 did not have LNM. A taller-than-wide shape and T3 stage on US were associated with ETE. Female sex, total thyroidectomy, and T3 stage on US were associated with LNM. When B-mode US and SWE were combined, there was no improvement in diagnostic performance.Combination of SWE and B-mode US findings is not useful for predicting ETE and LNM status in PTC patients.

A national database analysis for factors associated with thyroid cancer occurrence.

In order to analyze the associations between thyroid cancer and environmental factors, we analyzed the national sample cohort representative of the entire population provided by the Korean National Health Insurance Service database record from 2006 to 2015. The cohort was categorized according to age, body mass index, income, residential areas, frequency of exercise, frequency of alcohol drinking, diet, presence or absence of hyperthyroidism, presence or absence of hypothyroidism, and smoking data. Age ≥ 55 years (HR 0.68, 95% CI 0.53-0.88), lower income (0.57, 0.40-0.80), and current smoking (0.69, 0.55-0.85) were associated with lower thyroid cancer occurrence among men. Body mass index (BMI) ≥ 25 kg/m2 (1.51, 1.26-1.82), higher income (1.44, 1.19-1.76), urban residence (1.24, 1.03-1.49), and presence of hypothyroidism (3.31, 2.38-4.61) or hyperthyroidism (2.46, 1.75-3.46) were associated with higher thyroid cancer occurrence among men. Age ≥ 55 years (0.63, 0.56-0.71), moderate alcohol drinking (0.87, 0.77-0.99), and current smoking (0.56, 0.37-0.85) were associated with lower thyroid cancer occurrence among women. BMI ≥ 25 kg/m2 (1.41, 1.26-1.57), frequent exercise (1.21, 1.07-1.36), higher income (1.18, 1.06-1.32), urban residence (1.17, 1.06-1.29), and presence of hypothyroidism (1.60, 1.40-1.82) or hyperthyroidism (1.38, 1.19-1.61) were associated with higher thyroid cancer occurrence among women. In conclusion, age ≥ 55 years and current smoking were associated with lower thyroid cancer occurrence, while BMI ≥ 25 kg/m2, higher income, urban residence, hypothyroidism, and hyperthyroidism were associated with higher occurrence in both men and women.

Diagnostic Performance of a Combination of Shear Wave Elastography and B-Mode Ultrasonography in Differentiating Benign From Malignant Thyroid Nodules.

OBJECTIVES: This study was conducted to compare clinicopathologic and radiologic factors between benign and malignant thyroid nodules and to evaluate the diagnostic performance of shear wave elastography (SWE) combined with B-mode ultrasonography (US) in differentiating malignant from benign thyroid nodules. METHODS: This retrospective study included 92 consecutive patients with 95 thyroid nodules examined on B-mode US and SWE before US-guided fine-needle aspiration biopsy or surgical excision. B-mode US findings (composition, echogenicity, margin, shape, and calcification) and SWE elasticity parameters (maximum [Emax], mean, minimum, and nodule-to-normal parenchymal ratio of elasticity) were reviewed and compared between benign and malignant thyroid nodules. The diagnostic performance of B-mode US and SWE for predicting malignant thyroid nodules was analyzed. The optimal cutoff values of elasticity parameters for identifying malignancy were determined. Diagnostic performance was compared between B-mode US only, SWE only, and the combination of B-mode US with SWE. RESULTS: On multivariate logistic regression analysis, age (odds ratio [OR], 0.90; P=0.028), a taller-than-wide shape (OR, 11.3; P=0.040), the presence of calcifications (OR, 15.0; P=0.021), and Emax (OR, 1.22; P=0.021) were independent predictors of malignancy in thyroid nodules. The combined use of B-mode US findings and SWE yielded improvements in sensitivity, the positive predictive value, the negative predictive value, and accuracy compared with the use of B-mode US findings only, but with no statistical significance. CONCLUSION: When SWE was combined with B-mode US, the diagnostic performance was better than when only B-mode US was used, although the difference was not statistically significant.

Characterisation of fungal contamination sources for use in quality management of cheese production farms in Korea.

OBJECTIVE: This study was conducted to determine the composition and diversity of the fungal flora at various control points in cheese ripening rooms of 10 dairy farms from six different provinces in the Republic of Korea. METHODS: Floor, wall, cheese board, room air, cheese rind and core were sampled from cheese ripening rooms of ten different dairy farms. The molds were enumerated using YM petrifilm, while isolation was done on yeast extract glucose chloramphenicol agar plates. Morphologically distinct isolates were identified using sequencing of internal transcribed spacer region. RESULTS: The fungal counts in 8 out of 10 dairy farms were out of acceptable range, as per hazard analysis critical control point regulation. A total of 986 fungal isolates identified and assigned to the phyla Ascomycota (14 genera) and Basidiomycota (3 genera). Of these Penicillium, Aspergillus, and Cladosporium were the most diverse and predominant. The cheese ripening rooms was overrepresented in 9 farms by Penicillium (76%), while Aspergillusin a single farm. Among 39 species, the prominent members were Penicillium commune, P. oxalicum, P. echinulatum, and Aspergillus versicolor. Most of the mold species detected on surfaces were the same found in the indoor air of cheese ripening rooms. CONCLUSION: The environment of cheese ripening rooms persuades a favourable niche for mold growth. The fungal diversity in the dairy farms were greatly influenced by several factors (exterior atmosphere, working personnel etc.,) and their proportion varied from one to another. Proper management of hygienic and production practices and air filtration system would be effective to eradicate contamination in cheese processing industries.

Comparison of postoperative outcomes between bilateral axillo-breast approach-robotic thyroidectomy and transoral robotic thyroidectomy.

BACKGROUND: The use of robotic systems for thyroidectomy has increased as it enables more diverse approaches than the conventional open method. The aim of this study was to compare the clinical outcomes of Transoral Robotic Thyroidectomy (TORT) and Bilateral Axillo-Breast Approach-Robotic Thyroidectomy (BABA-RT). METHODS: This study was designed as a retrospective study. The included patients who underwent surgery by BABA-RT or TORT approach in our facility between 2008 and 2018. All surgeries were performed by one surgeon. Total thyroidectomy with central node dissection (CND) was performed only if tumors were >4 cm and had extrathyroidal extension, clinically apparent lymph node or distant metastases. In all other cases, lobectomy ± CND was performed. RESULTS: The group treated with TORT comprised 248 patients and the group that underwent BABA-RT had 316 patients. The number of retrieved lymph node (LN) was higher in the TORT group (4.9±4.4 vs. 4.2±4.9; P=0.01). There were no significant differences between the TORT and BABA-RT groups in concerns to the location of the tumor. Postoperative hospital stay was also shorter in the TORT group when compared with the BABA-RT group (2.8±0.90 vs. 3.4±0.97 days, P=0.012). Operative time was significantly shorter in the TORT group (204.11±40.19 vs. 243.78±57.16 min, P<0.01). CONCLUSIONS: When comparing a total of 248 patients treated with TORT versus 316 with BABA-RT. TORT not only has advantages in better cosmetic outcomes with minimized postoperative scars, but also shows comparable, or even superior, surgical outcomes with shorter operation time than the BABA-RT procedure.

Comparison Study of Robotic Thyroidectomies Through a Bilateral Axillo-Breast Approach and a Transoral Approach.

Introduction: Oncological and surgical safeties are pivotal issues of cancer operations. Robotic thyroidectomy adds cosmetic advantage to those safeties. We have performed bilateral axillo-breast approach robotic thyroidectomies (BABART) since 2009 and recently started transoral robotic thyroidectomy (TORT) in 2017. This study aimed to compare the surgical outcomes of a single surgeon's initial TORT and BABART. Materials and Methods: We retrospectively collected data of 103 patients who underwent robotic thyroid lobectomy for papillary thyroid cancer and analyzed the first 14 and 56 cases of TORT and BABART, respectively, after propensity score matching. The surgeon performed 224 BABARTs before starting TORT. Results: There were no significant differences between the BABART and TORT groups in mean age (40.02 ± 9.37 versus 38.69 ± 9.21 years, respectively; P = .7520), sex distribution (P = .3697), mean body mass index (23.60 ± 4.31 versus 23.87 ± 2.45 kg/m2, respectively; P = .4737), and tumor size (0.75 ± 0.35 versus 0.76 ± 0.29 cm, respectively; P = .9969). The TORT group had a longer operative time than the BABART group by 78.04 minutes (P < .0001). The visual analog scale pain scores on postoperative day (POD) 2 and POD 3 were higher in the TORT than the BABART group by 0.59 and 0.77, respectively (P = .0227 and .0119, respectively). The number of retrieved lymph nodes and unintended parathyroidectomies was similar in the two groups. There were no severe complications such as tracheal injury, transection of recurrent laryngeal nerve, or surgical site infection. Conclusion: Our study suggests that both BABART and TORT are safe and feasible during the initial period. TORT can be undertaken without any adverse event if the operator is experienced with other robotic thyroidectomy. The patients may choose the surgical approach based on their preference.

Inhibition of tamoxifen's therapeutic effects by emodin in estrogen receptor-positive breast cancer cell lines.

PURPOSE: This study was aimed to investigate the combination effect of endoxifen and emodin on estrogen receptor (ER) positive breast cancer cell lines and to explain the mechanism of the combination effect. METHODS: We conducted this study on MCF-7 (ER+/human epidermal growth factor receptor-2 [HER2]-), T47D (ER+/HER2-), ZR-75-1 (ER+/HER2+), and BT474 (ER+/HER2+) cell lines, which confirmed combination effect of endoxifen and emodin. Optimal concentrations for combination were determined to study the effects on proliferation of MCF-7 and ZR-75-1 cells. Analysis of the combination effect was carried out in the CompuSyn software. The combination of downstream mechanisms, and combined effects of other similar compounds were tested on the MCF-7 and ZR 75-1 cell lines. Protein expression was confirmed by western blot. RESULTS: The combination of endoxifen and emodin had antagonistic effects on MCF-7 and ZR-75-1cell lines (combination index > 1). We validated the antagonistic effect in T47D and BT474 cell lines. During the combined treatment, the results showed elevated amounts of cyclin D1 and phosphorylated extracellular signal-regulated kinase (pERK). Analysis of drug interactions showed antagonistic effect between endoxifen and chemical compounds similar to emodin, such as chrysophanol or rhein, in MCF-7 and ZR-75-1 cells. CONCLUSION: Addition of emodin attenuated tamoxifen's treatment effect via cyclin D1 and pERK up-regulation in ER-positive breast cancer cell lines.

Postthyroidectomy obesity in a Korean population: does the extent of surgery matter?

PURPOSE: The purpose of this study was to investigate the prevalence of postthyroidectomy obesity, and the relationship between the extent of thyroidectomy and obesity. METHODS: A survey conducted at an outpatient clinic from June to October 2014 and retrospective charts for patients undergoing thyroidectomy at Konkuk University Medical Centers from June 2009 to December 2013 were reviewed. We compared clinical characteristics and pre- and postoperative obesity-related factors in 227 patients who underwent total thyroidectomy or lobectomy. RESULTS: Patients included 39 males and 188 females with a mean age of 46.0 ± 11.0 years; the mean follow-up period was 23.9 ± 16.7 months, and 90 of the 227 patients showed postthyroidectomy obesity. In effect of operative extent on postoperative obesity, patients who underwent TT (48.2 years) than those who underwent lobectomy (43.4 years). TT group had longer follow-up and the frequency of menopause was higher than in the lobectomy group. No differences in postthyroidectomy obesity, body weight change, or body mass index (BMI), change among 2 groups. The predictors of postthyroidectomy obesity were older age, female, heavy alcohol consumption (P = 0.029), higher preoperative BMI (P < 0.001), larger postoperative weight gain (P = 0.024), and larger BMI change. However, the extent of thyroidectomy did not affect postthyroidectomy obesity. Preoperative BMI (P < 0.001) and heavy alcohol consumption (P = 0.03) were independent factors of postthyroidectomy obesity. CONCLUSION: The extent of thyroidectomy does not affect postthyroidectomy obesity. Preoperative BMI and heavy alcohol consumption are risk factors for postthyroidectomy obesity. Studies are needed to suggest preoperative life style modification to prevent postthyroidectomy obesity.

Expression and Purification of Extracellular Solute-Binding Protein (ESBP) in Escherichia coli, the Extracellular Protein Derived from Bifidobacterium longum KACC 91563.

Bifidobacterium longum KACC 91563 secretes family 5 extracellular solute-binding protein via extracellular vesicle. In our previous work, it was demonstrated that the protein effectively alleviated food allergy symptoms via mast cell specific apoptosis, and it has revealed a therapeutic potential of this protein in allergy treatment. In the present study, we cloned the gene encoding extracellular solute-binding protein of the strain into the histidine-tagged pET-28a(+) vector and transformed the resulting plasmid into the Escherichia coli strain BL21 (DE3). The histidine-tagged extracellular solute-binding protein expressed in the transformed cells was purified using Ni-NTA affinity column. To enhance the efficiency of the protein purification, three parameters were optimized; the host bacterial strain, the culturing and induction temperature, and the purification protocol. After the process, two liters of transformed culture produced 7.15 mg of the recombinant proteins. This is the first study describing the production of extracellular solute-binding protein of probiotic bacteria. Establishment of large-scale production strategy for the protein will further contribute to the development of functional foods and potential alternative treatments for allergies.

Hesperidin Shows Protective Effects on Renal Function in Ischemia-induced Acute Kidney Injury (Sprague-Dawley Rats).

OBJECTIVE: Hesperidin is a well-known flavanone glycoside copiously found in sweet orange and lemon, which was recently reported to possess significant anti-inflammatory, analgesic, antifungal, antiviral, antioxidant, and anticancer activities. Ischemia-reperfusion (I/R) injury is a major problem after renal transplantation. Furthermore, inflammatory responses to I/R exacerbate the resultant renal injury. In the present study, we investigated whether hesperidin exhibits renoprotective effects against I/R-induced acute kidney injury in a rat model. METHODS: We fed Sprague-Dawley rats either hesperidin (100 mg/kg/d) or saline. One week later, ischemia was induced by bilateral renal pedicle occlusion for 30 minutes followed by reperfusion. The rats were randomly divided into 3 groups, which were treated as follows: 1. the sham operated group; 2. the I/R group; 3. the I/R-hesperidin group RESULTS: Compared to the sham group, the I/R group had higher expression of blood urea nitrogen and serum creatinine and lower expression of catalase, superoxide dismutase, glutathione peroxidase, antioxidants, and nitric oxide. Compared to the I/R group, the I/R-hesperidin group had higher expression of catalase, superoxide dismutase, glutathione peroxidase, antioxidant, and nitric oxide and lower expression of blood urea nitrogen and serum creatinine. CONCLUSIONS: Hesperidin improved acute renal I/R injury through its antioxidant effects. These findings suggest that hesperidin is a potential therapeutic agent for acute ischemia-induced renal damage.

Upregulation of SLC2A3 gene and prognosis in colorectal carcinoma: analysis of TCGA data.

BACKGROUND: Upregulation of SLC2A genes that encode glucose transporter (GLUT) protein is associated with poor prognosis in many cancers. In colorectal cancer, studies reporting the association between overexpression of GLUT and poor clinical outcomes were flawed by small sample sizes or subjective interpretation of immunohistochemical staining. Here, we analyzed mRNA expressions in all 14 SLC2A genes and evaluated the association with prognosis in colorectal cancer using data from the Cancer Genome Atlas (TCGA) database. METHODS: In the present study, we analyzed the expression of SLC2A genes in colorectal cancer and their association with prognosis using data obtained from the TCGA for the discovery sample, and a dataset from the Gene Expression Omnibus for the validation sample. RESULTS: SLC2A3 was significantly associated with overall survival (OS) and disease-free survival (DFS) in both the discovery sample (345 patients) and validation sample (501 patients). High SLC2A3 expression resulted in shorter OS and DFS. In multivariate analyses, high SLC2A3 levels predicted unfavorable OS (adjusted HR 1.95, 95% CI 1.22-3.11; P = 0.005) and were associated with poor DFS (adjusted HR 1.85, 95% CI 1.10-3.12; P = 0.02). Similar results were found in the discovery set. CONCLUSION: Upregulation of the SLC2A3 genes is associated with decreased OS and DFS in colorectal cancer patients. Therefore, assessment of SLC2A3 gene expression may useful for predicting prognosis in these patients.

Prognostic nomogram of hypoxia-related genes predicting overall survival of colorectal cancer-Analysis of TCGA database.

Hypoxia-related gene (HRG) expression is associated with survival outcomes of colorectal cancer (CRC). Our aim was developing a nomogram predicting CRC overall survival (OS) with HRGs and clinicopathological factors. The Cancer Genome Atlas (TCGA) database was used as discovery cohort and two Gene Expression Omnibus databases (GSE39582 and GSE41258) served as validation cohorts. A genetic risk score model prognosticating OS was developed using mRNA expression level of HRGs. Nomogram predicting OS was developed using genetic risk score model and clinicopathological variables. The genetic risk score model included four HRGs (HSPA1L, PUM1, UBE2D2, and HSP27) and successfully prognosticated OS of discovery and two validation cohorts (p < 0.001 for TCGA discovery set, p < 0.003 for the GSE39582 and p = 0.042 for the GSE41258 datasets). Nomogram included genetic risk score, age, and TNM stage. Harrell's concordance indexes of the nomogram were higher than those of TNM stage alone in the discovery set (0.77 vs. 0.69, p < 0.001), GSE39582 (0.65 vs. 0.63, p < 0.001), and GSE41258 datasets (0.78 vs. 0.77, p < 0.001). Our nomogram successfully predicted OS of CRC patients. The mRNA expression level of the HRGs might be useful as an ancillary marker for prognosticating CRC outcome.

Colorectal Cancer Prognosis is Not Associated with BRAF and KRAS Mutations-A STROBE Compliant Study.

BACKGROUND: We investigated the associations between v-Raf murine sarcoma viral oncogene homolog B1 (BRAFV600E, henceforth BRAF) and v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS) mutations and colorectal cancer (CRC) prognosis, using The Cancer Genome Atlas (TCGA) and the Gene Expression Omnibus (GSE39582) datasets. MATERIALS AND METHODS: The effects of BRAF and KRAS mutations on overall survival (OS) and disease-free survival (DFS) of CRC were evaluated. RESULTS: The mutational status of BRAF and KRAS genes was not associated with overall survival (OS) or DFS of the CRC patients drawn from the TCGA database. The 3-year OS and DFS rates of the BRAF mutation (+) vs. mutation (-) groups were 92.6% vs. 90.4% and 79.7% vs. 68.4%, respectively. The 3-year OS and DFS rates of the KRAS mutation (+) vs. mutation (-) groups were 90.4% vs. 90.5% and 65.3% vs. 73.5%, respectively. In stage II patients, however, the 3-year OS rate was lower in the BRAF mutation (+) group than in the mutation (-) group (85.5% vs. 97.7%, p <0.001). The mutational status of BRAF genes of 497 CRC patients drawn from the GSE39582 database was not associated with OS or DFS. The 3-year OS and DFS rates of BRAF mutation (+) vs. mutation (-) groups were 75.7% vs. 78.9% and 73.6% vs. 71.1%, respectively. However, KRAS mutational status had an effect on 3-year OS rate (71.9% mutation (+) vs. 83% mutation (-), p = 0.05) and DFS rate (66.3% mutation (+) vs. 74.6% mutation (-), p = 0.013). CONCLUSIONS: We found no consistent association between the mutational status of BRAF nor KRAS and the OS and DFS of CRC patients from the TCGA and GSE39582 databases. Studies with longer-term records and larger patient numbers may be necessary to expound the influence of BRAF and KRAS mutations on the outcomes of CRC.