Effect of saikosaponins-b2 on migration of HepG2 cells and expressions of angiogenesis-related proteins / 中国药理学与毒理学杂志

OBJECTIVE: To investigate the effect of saikosaponins-b2 (SS-b2) on the migration of HepG2 cells, and to explore its potential molecular mechanism. METHODS: The effect of SS-b2 on the viability of HepG2 cells was detected by MTT method before the cells were divided into normal control group, SS-b2 (15, 30 and 60 mg·L-1) groups and positive control [doxorubicin (DOX) 2 mg·L-1] group according to the results of MTT. Wound-healing assay was performed to observe the influence of SS-b2 on the migration of HepG2 cells. The protein expressions of vascular endothelial growth factor (VEGF), matrix metalloproteinase 2 (MMP-2), MMP-9, hypoxia inducible factor-lot (HIF-1α) and the phosphorylation of extracellular regulated protein kinase 1/2 (ERK1/2) in HepG2 cells were measured by Western blotting. RESULTS: The wound-healing rate was 74.71%, 72.50% and 66.82% after treatment with SS-b2 15, 30 and 60 mg·L-1for 48 h, respectively. The wound-healing rate decreased significantly with the increae of SS-b2 doses. Compared with the normal control group, the wound-healing rate of SS-b2 30 and 60 mg·L-1was significantly reduced (P<0.05). The expressions of VEGF and HIF-1α in HepG2 cells were significantly decreased in SS-b2 groups (P<0.05, P<0.01). SS-b2 60 mg·L-1could reduce the protein expressions of MMP-2 and MMP-9 and the phosphorylation of ERK1/2 significantly (P< 0.05, P<0.01). The positive control group could significantly reduce the expressions of the above four proteins and the phosphorylation of ERK1/2 (P<0.05, P<0.01). Compared with the positive control group, the protein expressions of VEGF and MMP-2 were significantly down-regulated in 60 mg·L-1SS-b2 group (PO.05, P<0.01), but there was no significant difference in the expression of HIF-1α between SS-b2 groups and the positive control group. However, the down-regulation effect of SS-b2 on MMP-9 protein expression was not so significant as in the positive control group (P<0.01). The effect of SS-b2 60 mg·L-1on the phosphorylation of ERK1/2 was similar to that of the positive control group. CONCLUSION: SS-b2 can inhibit the migration of HepG2 cells, which may be related to the inhibition of angiogenesis-related proteins.

[Effects of Different Levels of Nitrogen Fertilization on Soil Respiration Rates and Soil Biochemical Properties in an Alfalfa Grassland].

Understanding the effects of different levels of nitrogen fertilizer applications on soil respiration rates and soil biochemical properties is of great importance for providing a theoretical basis for accurate assessments of the soil respiration intensity and carbon recycling in grassland ecosystems. A field experiment was performed from April 2017 to March 2018, in which four different levels of nitrogen applications were investigated, including 0 kg·hm-2 (N0), 60 kg·hm-2 (N1), 120 kg·hm-2 (N2), and 180 kg·hm-2 (N3). The seasonal changes in the soil respiration rate, soil temperature, and soil moisture in the alfalfa grassland under different levels of nitrogen applications were observed, and soil biochemical characteristics were observed after each harvest in the growing season. The results showed that soil respiration rate of the alfalfa grassland displayed significant seasonal variation under different nitrogen levels. In particular, the soil respiration rate reached a peak during the last 10-day period of July and then decreased to the minimum in mid-December. During the growing season of alfalfa, the soil respiration rate of the alfalfa grassland increased with the increases in the nitrogen application rate. The mean soil respiration rates of the N1, N2, and N3 treatments were 0.97, 1.04, and 1.07 g·(m2·h)-1, respectively, and these values were 10.2%, 18.2%, and 21.6% greater than that of N0[0.88 g·(m2·h)-1], respectively. The results from ANOVA testing indicated that nitrogen applications had no significant effect on the soil respiration rate during the non-growing season of alfalfa (P>0.05). According to the statistical analysis, the soil respiration rate had a significant exponential positive relationship with soil temperature during the growing season, non-growing season, and entire year of alfalfa grassland observations under different nitrogen application rates (P<0.01); the coefficients of determination were ranked as follows:growing season (0.46-0.62) < non-growing season (0.66-0.76) < whole year (0.80-0.86). Soil temperature (T) and soil moisture (W) interacted with each other and ultimately affected the soil respiration (RS), and by using a two-factor linear model of soil temperature and soil moisture, a better fit was obtained for the change in the soil respiration rate. Both of the two factors explained 68%-80% of the variation in the seasonal soil respiratory rate during the growing season of alfalfa. Nitrogen fertilization decreased the soil pH and available phosphorus content (AP) to varying degrees, but it increased the available potassium (AK), soil organic matter (SOM), and soil urease (URE) and invertase activity (INV). Total nitrogen (TN) and available nitrogen (AN) showed different trends under different nitrogen levels. The TN and AN contents increased considerably in soils; however, when the nitrogen rate was higher than N2 (120 kg·hm-2), TN and AN decreased with the increases in the nitrogen application rate. According to the correlation matrix analysis between soil respiration and soil biochemical properties during the growth period of alfalfa, data showed that the soil respiration rate (RS) was significantly and negatively correlated with soil pH (P<0.01), and it was significantly and positively correlated with soil TN and URE (P<0.01). Simultaneously, there was a significant positive correlation between the soil respiration rate (RS) and SOM (P<0.05), and there was a significant negative correlation with INV (P<0.05). The soil nutrient and enzyme activities of the alfalfa grassland explained the variations in the soil respiration rate under different nitrogen application levels to varying degrees.

Effects of saikosaponin b_2 on inflammation and energy metabolism in mice with acute liver injury induced by LPS/GalN / 中国中药杂志

To study the effects of saikosaponin b2( SS-b2) on inflammatory factors and energy metabolism against lipopolysaccharide/galactosamine( LPS/Gal N) induced acute liver injury in mice. Mice were randomly divided into normal group( equal amount of normal saline),model group( 100 g·kg~(-1) LPS and 400 mg·kg~(-1) Gal N),low,medium,high dose group of SS-b2( SS-b25,10,20 mg·kg~(-1)·d-1) and positive control group( dexamethasone,10 mg·kg~(-1)). All of the groups except for the normal group were treated with LPS/Gal N though intraperitoneally injection to establish the acute liver injury model. The organ indexes were calculated. The levels of serum transaminases( ALT and AST) and the activities of ATPase( Na+-K+-ATPase,Ca2+-Mg2+-ATPase) in liver were detected. The activity of tumor necrosis factor-α( TNF-α),interleukin-1β( IL-1β) and interleukin-6( IL-6) were determined by the enzyme-linked immunosorbent assay( ELISA). The contents of lactate dehydrogenase( LDH) in liver were determined by micro-enzyme method. HE staining was used to observe the histopathological changes of the liver. Histochemical method was used to investigate the protein expression of liver lactate dehydrogenase-A( LDH-A). The protein expressions of Sirt-6 and NF-κB in the liver were detected by Western blot. According to the results,compared with the model group,there were significant changes in organ indexes in the high-dose group of SS-b2( P<0. 05). The level of ALT,AST,TNF-α,IL-1β,IL-6 and the activities of LDH in serum of mice with liver injury were significantly reduced in the medium and high dose groups of SS-b2( P<0. 01). With the increase of the concentration of SS-b2,the range of hepatic lesions and the damage in mice decreased. The activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase in liver of mice were significantly enhanced in each dose group( P<0. 01). The expression of NF-κB in liver tissues was significantly down-regulated in the medium and high dose group( P<0. 01). Meanwhile,the expression of Sirt-6 protein in the liver of mice with acute liver injury was significantly increased in each dose group( P<0. 01).In summary,SS-b2 has a significant protective effect on LPS/Gal N-induced acute liver injury in mice,which may be related to the down-regulation of NF-κB protein expression and up-regulation of Sirt-6 protein expression to improve inflammatory injury and energy metabolism.

Survival Improvement in Korean Breast Cancer Patients Due to Increases in Early-Stage Cancers and Hormone Receptor Positive/HER2 Negative Subtypes: A Nationwide Registry-Based Study / 한국유방암학회지

PURPOSE: The aim of this study was to investigate whether the observed changes over time in the survival rates vary according to the intrinsic subtypes of breast cancer diagnosed. METHODS: Data from 46,320 breast cancer patients in the Korean Breast Cancer Registry who underwent surgery between 1999 and 2006 were reviewed. Among them, results from 25,887 patients with available data about the status of estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2 (HER2) were analyzed. Patients were classified into two cohorts according to the year in which they underwent surgery: 1999-2002 and 2003-2006. RESULTS: The patients treated in the latter time period showed significantly better overall survival (OS) compared with those in the former period when adjusted for follow-up duration. The proportion of hormone receptor+/HER2-subtype and stage I breast cancer were significantly higher in the latter period (47.4% vs. 54.6%, p<0.001; 31.0% vs. 39.6%, p<0.001, respectively). Improvement in OS between the former and latter periods was seen in all subtypes of breast cancer, including triple-negative cancers (all p-values <0.001 in univariate and multivariate analyses). CONCLUSION: Improvement in survival in Korean breast cancer patients over the study years is being observed in all subtypes of breast cancer, implying that increases in both early-stage detection and the proportion of less aggressive cancers contribute to this improvement.

Low Rates of Additional Cancer Detection by Magnetic Resonance Imaging in Newly Diagnosed Breast Cancer Patients Who Undergo Preoperative Mammography and Ultrasonography / 한국유방암학회지

PURPOSE: We evaluated the efficacy of breast magnetic resonance imaging (MRI) for detecting additional malignancies in breast cancer patients newly diagnosed by breast ultrasonography and mammography. METHODS: We retrospectively reviewed the records of 1,038 breast cancer patients who underwent preoperative mammography, bilateral breast ultrasonography, and subsequent breast MRI between August 2007 and December 2010 at single institution in Korea. MRI-detected additional lesions were defined as those lesions detected by breast MRI that were previously undetected by mammography and ultrasonography and which would otherwise have not been identified. RESULTS: Among the 1,038 cases, 228 additional lesions (22.0%) and 30 additional malignancies (2.9%) were detected by breast MRI. Of these 228 lesions, 109 were suspected to be malignant (Breast Imaging-Reporting and Data System category 4 or 5) on breast MRI and second-look ultrasonography and 30 were pathologically confirmed to be malignant (13.2%). Of these 30 lesions, 21 were ipsilateral to the main lesion and nine were contralateral. Fourteen lesions were in situ carcinomas and 16 were invasive carcinomas. The positive predictive value of breast MRI was 27.5% (30/109). No clinicopathological factors were significantly associated with additional malignant foci. CONCLUSION: Breast MRI was useful in detecting additional malignancy in a small number of patients who underwent ultrasonography and mammography.

Validation of a Scoring System for Predicting Malignancy in Patients Diagnosed with Atypical Ductal Hyperplasia Using an Ultrasound-Guided Core Needle Biopsy / 한국유방암학회지

PURPOSE: The need for surgical excision in patients with ultrasound-guided core needle biopsy (CNB)-diagnosed atypical ductal hyperplasia (ADH) remains an issue of debate. The present study sought to validate a scoring system (the U score, for underestimation) that we have previously developed for predicting malignancy in CNB-diagnosed ADH. METHODS: The study prospectively enrolled 85 female patients with CNB-diagnosed ADH who underwent subsequent surgical excision. Underestimation was defined as a surgical specimen having malignant foci. RESULTS: The overall underestimation rate was 37% (31/85). Multivariate analysis showed that a clinically palpable mass, microcalcification on imaging, size >15 mm and a patient age of > or =50 years were independently associated with underestimation. When applied to the scoring system, the validation score was significant (p<0.001; area under the curve, 0.852). No patient with a U score <3.5 had an underestimated lesion. CONCLUSION: The present study successfully validated the efficacy of our scoring system for predicting malignancy in CNB-diagnosed ADH. A U score of < or =3.5 indicates that surgical excision may not be necessary.

The Impact of Appendiceal CT on Patients with Suggestive Appendicitis

PURPOSE: This study was performed to evaluate the contribution of appendiceal computed tomography (CT) in the more accurate diagnosis of acute appendicitis and decrease in the rate of negative appendectomies. METHODS: Between May and August 2005, 146 patients with right lower quadrant (RLQ) abdominal pain and tenderness were diagnosed using appendiceal CT. The appendiceal CT scans were performed with contrast media in the abdominal and pelvic areas. 7 mm slice scans were taken both before and after the contrast media injection, with a time delay. The control group was comprised of 99 patients, who were also checked by abdominal CT or ultrasonography, between December 2004 and April 2005, with the data analyzed retrospectively. RESULTS: In the study group, 85 of the 146 cases were diagnosed with acute appendicitis on appendiceal CT, and had undergone an appendectomy, including 42 men (mean age 39.9 yr) and 43 women (mean age 44.6 yr). The sensitivity and specificity of appendiceal CT in acute appendicitis were 95.3 and 98.4%, respectively. There were 4 (4.7%) negative appendectomy cases; 2 in men and 2 in women. The control group conprised of 47 men and 52 women. There were 13 (13.1%) negative appendectomy cases; 4 in men and 9 in women. CONCLUSION: The routine use of appendiceal CT, in patients with RLQ abdominal pain and tenderness, resulted in a significant decrease in the negative appendectomy rate.

Direct Comparison of PHS(R) and Perfix(R) Herniorrhaphy under Local Anesthesia

PURPOSE: The aims of this study were to evaluate the feasibility of local anesthesia in tension-free herniorrhaphy, using prosthetic mesh, and to directly compare PHS(R) and Perfix(R) herniorrhaphy under local anesthesia. METHODS: Patients with a groin hernia, who underwent tension-free herniorrhaphy (n = 107) under local anesthesia between March 2003 and February 2006, were included. PHS(R) (n = 63) and Perfix(R) (n = 44) meshes were randomly used, with no difference between the PHS(R) and Perfix(R) groups in relation to mean age, gender, number of combined diseases, body mass index (BMI), recurred hernia and types of hernia. RESULTS: Intraoperative analgesics and/or sedatives were used in 55 patients (51.4 %) where local anesthesia was insufficient. In a univariate analysis, the additional use of intraoperative analgesics and/or sedatives was related to the patient's age and BMI. The patients at an older age and with a lower BMI were more tolerant to local anesthesia. However, only the BMI was found to be a statistically significant factor from the multivariate analysis. There was no significant difference between the PHS(R) and Perfix(R) groups on the additional use of intraoperative analgesics and/or sedatives, the use of postoperative analgesics, length of hospital stay, complication and recurrence (P > 0.05). CONCLUSION: With tension-free herniorrhaphy using the PHS(R) or Perfix(R) mesh, local anesthesia was acceptable and securable, regardless of the mesh type used. Among these patients, those at an older age and with a lower BMI were more tolerant to local anesthesia.

Multiple acyl-CoA desaturase-encoding transcripts in pheromone glands of Helicoverpa assulta, the oriental tobacco budworm.

Seven desaturase cDNAs were isolated from pheromone glands of Helicoverpa assulta, a moth producing a sex pheromone blend with high Z9-16:Ald and low Z11-16:Ald, opposite to what is found in other heliothine moths such as Helicoverpa zea. Six of the seven sequences map onto recently defined lepidopteran desaturase sequence lineages and the other is orthologous to a desaturase sequence previously reported only in H. zea. The levels of desaturase-encoding transcripts in pheromone glands were determined and the three most abundant ones were functionally expressed in a desaturase-deficient mutant strain of Saccharomyces cerevisiae. The HassNPVE transcript, shown to encode a delta9 desaturase producing more Z9-18:Acid than Z9-16:Acid, was the most abundant, followed by the HassKPSE transcript, shown to encode a delta9 desaturase producing more Z9-16:Acid than Z9-18:Acid, and by the HassLPAQ transcript, shown to encode a delta11 desaturase producing only Z11-16:Acid. Thus, the relative amounts of transcripts encoding two delta9 desaturases and a single delta11 desaturase in H. assulta pheromone glands were consistent with the relative amounts of unsaturated fatty acid precursors required to produce the major and minor sex pheromone components of this species. Desaturase transcript levels in pheromone glands were also found to be as high during scotophase as during light phase, when pheromone production ceases. The other four transcripts were present at extremely low levels in H. assulta pheromone glands and the functional roles of their encoded desaturase-homologous proteins could not be determined.