Subchronic oral toxicity study of rhubarb extract in Sprague-Dawley rats.

OBJECTIVE: The study was primarily used to evaluate subchronic oral toxicity of rhubarb extract. METHODS: The rhubarb extract was orally administered to rats at doses of 0.00, 0.65, 1.62 and 4.05 g/kg BW/day for 13 weeks with a recovery period of 4 weeks. The weight and the relative organ weight of the kidney in the 0.65 g/kg BW group were significantly increased but no significant changes were seen in renal histopathology. When the rats received rhubarb extract at 1.62 g/kg BW or above, the relative weight of the spleen and kidney were significantly increased; the kidney was also swollen and black with hydronephrosis. Histologic examination showed that there was an obvious increase in pigment deposition in renal tubular epithelial cells. No toxic related changes were observed in the 0.65 g/kg BW group, even though organ weight was increased and relative ratio to body weight of kidney were observed at 0.65 g/kg BW dosage, no significant renal histopathologic changes were detected at this dose. Based on the current study conditions and results, the no observed adverse effect level (NOAEL) of rhubarb extract in rats is 0.65 g/kg BW/day.

Germline TP53 and MSH6 mutations implicated in sporadic triple-negative breast cancer (TNBC): a preliminary study.

BACKGROUND: Germline BRCA1/2 prevalence is relatively low in sporadic triple-negative breast cancer (TNBC). We hypothesized that non-BRCA genes may also have significant germline contribution to Chinese sporadic TNBC, and the somatic mutational landscape of TNBC may vary between ethnic groups. We therefore conducted this study to investigate germline and somatic mutations in 43 cancer susceptibility genes in Chinese sporadic TNBC. PATIENTS AND METHODS: Sixty-six Chinese sporadic TNBC patients were enrolled in this study. Germline and tumor DNA of each patient were subjected to capture-based next-generation sequencing using a 43-gene panel. Standard bioinformatic analysis and variant classification were performed to identify deleterious/likely deleterious germline mutations and somatic mutations. Mutational analysis was conducted to identify significantly mutated genes. RESULTS: Deleterious/likely deleterious germline mutations were identified in 27 (27/66, 40.9%) patients. Among the 27 patients, 9 (9/66, 13.6%) were TP53 carriers, 5 (5/66, 7.6%) were MSH6 carriers, and 5 (5/66, 7.6%) were BRCA1 carriers. Somatic mutations were identified in 64 (64/66, 97.0%) patients. TP53 somatic mutations occurred in most of the patients (45/66, 68.2%) and with highest mean allele frequency (28.1%), while NF1 and POLE were detected to have the highest mutation counts. CONCLUSIONS: Our results supported our hypotheses and suggested great potentials of TP53 and MSH6 as novel candidates for TNBC predisposition genes. The high frequency of somatic NF1 and POLE mutations in this study showed possibilities for clinical benefits from androgen-blockade therapies and immunotherapies in Chinese TNBC patients. Our study indicated necessity of multi-gene testing for TNBC prevention and treatment.

Subchronic toxicity of cerium nitrate by 90-day oral exposure in wistar rats.

This study evaluated the subchronic toxicity of cerium nitrate and determined the no observed adverse effect level (NOAEL) in Wistar rats. In accordance with the Organization for Economic Co-operation and Development guidelines, cerium nitrate was orally administered to Wistar rats by gavage at 0, 0.2, 75, 150, and 300 mg/kg bw/day for 90 days, followed by 28 days of recovery period in the 300 mg/kg bw/day and the control groups. The following parameters were evaluated: mortality, abnormalities, body weight, food consumption, hematology, serum biochemistry, urinanalysis, gross necropsy and histopathology. At the end of the treatment, several significant changes were observed in the 300 mg/kg bw/day groups: relatively decreased mean body weight of males, increased LYMPH%, RET% and decreased NEUT%, RBC of the females, increased ALT, AST and decreased ALB, T-Bil, CHO, CK, LDH of males. Significantly decreased T-Bil, CHO, CK and LDH were also observed in males of the 150 mg/kg bw/day group. Pathological examination revealed that the incidences of foreign body granulomatous lesions in lungs were higher in the 150 and 300 mg/kg bw/day groups as compared with the control group. These findings were attributed to unexpected gavage exposure because the granuloma exhibited a bronchiole-derived distribution. Taken together, the NOAEL of cerium nitrate in Wistar rats is set to be 75 mg/kg bw/day in the present study.

Therapeutic Effects of Bipolar Coagulation Forceps on Open Thyroid Surgery.

BACKGROUND: The aim was to compare the therapeutic effects of bipolar coagulation forceps, harmonic scalpel, and conventional thyroidectomy on open thyroid surgery. METHODS: A total of 527 patients who received open thyroid surgery in the Affiliated Drum Tower Hospital of Nanjing University Medical School between February 2013 and February 2016 were randomly divided into three groups: bipolar coagulation forceps, harmonic scalpel, and conventional thyroidectomy. There were no statistically significant differences in gender, age, disease constituents or mass diameter between the three groups. All surgeries were performed by the same surgeon. The surgical time, intraoperative blood loss, postoperative volume of drainage, postoperative hospital stay, and postoperative complications of the three surgical methods were compared. RESULTS: The bipolar coagulation forceps and harmonic scalpel groups were significantly superior to the conventional thyroidectomy group (p < 0.05) in terms of surgical time, intraoperative blood loss, postoperative volume of drainage, and postoperative hospital stay, but the first two groups had similar outcomes (p > 0.05). There were significant differences between the three groups in temporary recurrent laryngeal nerve palsy and temporary hypoparathyroidism, and the results of the bipolar coagulation forceps group were significantly better than those of the other two groups (p < 0.05). No significant differences were found in airway depression due to postoperative bleeding or irritating cough induced by superior laryngeal nerve palsy between the three groups (p > 0.05). None of the patients in the three groups suffered from permanent recurrent laryngeal nerve palsy or permanent hypoparathyroidism. CONCLUSIONS: The effects of bipolar coagulation forceps on open thyroid surgery exceeded those of the harmonic scalpel and conventional thyroidectomy. This method is worthy of promotion in clinical practice.

Extended one generation reproductive toxicity study and effect on gut flora of genetically modified rice rich in ß-carotene in wistar rats.

To evaluate the reproductive toxicity of gene modified rice generated by introducing phytoene synthase (Psy) and bacterial phytoene desaturase (CrtI) from maize and Erwinia uredovora, Wistar rats were allocated into 3 groups and fed with Psy and CrtI gene modified rice mixture diet (GM group), non-gene modified rice mixture diet (non-GM group), and AIN-93 diet (Blank control group) from parental generation (F0) to the offsprings (F1). GM rice, Heijinmi (HJM) and Non-GM rice, Heishuai (HS), were both formulated into diets at ratios of 73.5% and 75.5% according to the AIN93 diet for rodent animals, respectively. Relative to the non-GM group, no biologically relevant differences were observed in GM group rats concerning reproductive performance such as fertility rate, gestation rate, mean duration, hormone level, and reproductive organ pathology. The developmental parameters results were not significantly different from the non-GM group such as body weight, food consumption, developmental neurotoxicity, behavior, hematology, and serum chemistry. In terms of immunotoxicity, the IgG indicators of offspring from the GM group improved in contrast with the non-GM group. Additional gut flora analysis of F0 generation rats resulted as that the treatment elicited an increased gut microflora diversity of F0 rats. And no horizontal gene transfer of Psy and CrtI genes in rats fed a GM rice HJM diet. In conclusion, we found no adverse effects related to GM rice in the extended one-generation reproductive toxicity study, indicating that GM rice is a safe alternative for its counterpart rice regarding reproductive toxicity.

The physicochemical stability and in vivo gastrointestinal digestion of flaxseed milk: Implication of microwave on flaxseed.

The current study was to investigate how microwave on flaxseed affected the physicochemical stability and gastrointestinal digestion of oil bodies (OBs) in flaxseed milk. Flaxseed was subjected to moisture adjustment (30-35 wt%, 24 h), and microwave exposure (0-5 min, 700  W). Microwave treatment slightly weakened the physical stability of flaxseed milk indicated by Turbiscan Stability Index, but there were no visual phase separation during 21 days of storage at 4 °C. Upon microwave treatment, OBs experienced the layer-by-layer encapsulation into loose interface embedding by storage protein-gum polysaccharide complex from bulk phase, resulting in lower viscoelasticity of flaxseed milk. The OBs underwent earlier interface collapse and lipolysis during gastrointestinal digestion, followed by synergistic micellar absorption, faster chylomicrons transport within enterocytes of rats fed flaxseed milk. The accumulation of α-linolenic acid and synergistic conversion into docosapentaenoic and docosahexanoic acids in jejunum tissue were achieved accompanied by the interface remodeling of OBs in flaxseed milk.

Study on chronic toxicity of rhubarb extract in Sprague-Dawley rats.

Objective: The purpose of this study was to evaluate the safety of rhubarb extract. Methods: SD rats were treated with rhubarb extract at 0, 101, 405 and 1620 mg/kg/day for 52 weeks. food consumption and body weights were recorded. Blood and urine samples were collected for serum biochemical evaluation and urinalysis, and organ tissues were collected for histopathological examination. Results: The rats of 1620 mg/kg group developed diarrhea symptoms with dark brown loose stool after exposure; decreased body weight and increased food consumption were observed in the 1620 mg/kg and 405 mg/kg groups; urine WBC and NIT was significantly increased in the male and female rats of 1620 mg/kg group, and the urine pH was decreased in male rats of 1620 mg/kg group; renal tubular pigmentation was observed in the 1620 mg/kg group. Conclusion: The NOAEL of rhubarb extract on chronic toxicity (52 weeks) of Sprague-Dawley rats was 101 mg/kg in female and 94 mg/kg in male, and the LOAEL was 408 mg/kg in female and 381 mg/kg in male. The target organ of toxicity was the kidney, and the target cells was tubular epithelial cells.

Subchronic Toxicity of GmDREB3 Gene Modified Wheat in the Third Generation Wistar Rats.

The aim of the current study was to evaluate the subchronic toxicity of GmDREB3 gene modified wheat in the third generation rats. SPF Wistar rats were fed with transgenic wheat diet (Gm), parental wheat diet (Jimai22) and AIN-93 rodent diet (Control), respectively, for two generations, to produce the third generation rats which were used for this study. The selected fresh weaned offspring rats (20/sex/group) were given the same diet as their parents for 13 weeks. No toxicity-related changes were observed in rats fed with Gm diet in the following respects: clinical signs, body weights, body weight gains, food consumption, food utilization rate, urinalysis, hematology, serum biochemistry and histopathology. The results from the present study demonstrated that 13 weeks consumption of Gm wheat did not cause any adverse effects in the third generation rats when compared with the corresponding Jimai22 wheat.

Subchronic Oral Toxicity Study of Genetically Modified Rice Rich in ß-Carotene in Wistar Rats.

(1) Background: a hybrid black rice rich in ß-carotene carrying the psy and crtI genes (HJM) was evaluated in Wistar rats by a 90-day feeding study, aiming to assess its dietary safety. (2) Methods: the HJM rice and its parental line HS were included in rats' diets at levels of 73.5% and 75.5%, respectively. The AIN-93 diet was administered as a nutritional control. No adverse effects on animal behavior or weight gain were observed during the study. Blood samples were collected and analyzed, and standard hematological and biochemical parameters were compared. (3) Results: Some parameters were found to be significantly different, though they remained within the normal range for rats of this breed and age. In addition, upon sacrifice, various organs were weighed, and macroscopic and histopathological examinations were performed, with only minor changes to report. (4) Conclusions: HJM rice exhibited no adverse or toxic effects in Wistar rats in this 90-day study.

Two generation reproduction toxicity study of GmDREB3 gene modified wheat in Wistar rats.

To study reproductive toxicity of gene modified wheat generated by introducing DREB3 (drought response element binding protein 3) gene, Wistar rats of were allocated into 3 groups and fed with DREB3 gene modified wheat mixture diet (GM group), non-gene modified wheat mixture diet (Non-GM group) and AIN-93 diet (Control group) from parental generation (F0) to the second offspring (F2). GM wheat and Non-GM wheat, Jimai22, were both formulated into diets at a ratio of 69.55% according to AIN93 diet for rodent animals. Compared with non-GM group, no biologically related differences were observed in GM group rats with respect to reproductive performance such as fertility rate, gestation rate, mean duration, hormone level, reproductive organ pathology and developmental parameters such as body weight, body length, food consumption, neuropathy, behavior, immunotoxicity, hematology and serum chemistry. In conclusion, no adverse effect were found relevant to GM wheat in the two generation reproduction toxicity study, indicating the GM wheat is a safe alternative for its counterpart wheat regarding to reproduction toxicity.

miR-532-5p promotes breast cancer proliferation and migration by targeting RERG.

Aberrant expression of microRNAs (miRNAs/miRs) mediates the initiation and progression of breast cancer. Therefore, it is important to investigate the molecular mechanisms of miRNAs and their effects on breast cancer progression. In the present study, miR-532-5p was highly expressed in breast cancer tissues compared with normal tissues. In addition, expression of ras-related and estrogen-regulated growth inhibitor (RERG), a tumor suppressor in breast cancer, was negatively correlated with miR-532-5p expression. Inhibition of miR-532-5p significantly elevated RERG at both mRNA and protein levels and inactivated the mitogen-activated protein kinase (MAPK)/ERK signaling pathway. Overexpression of miR-532-5p decreased RERG expression and activated the MAPK/ERK signaling in breast cancer cell line MDA-MB-231. Bioinformatic analysis indicated that RERG 3'-untraslated region contained a putative binding site for miR-532-5p. Dual luciferase assay further validated RERG as a target gene of miR-532-5p. Notably, downregulation of miR-532-5p inhibited MDA-MB-231 cell proliferation and migration, which was partially attenuated upon RERG knockdown. In conclusion, the current study revealed an oncogenic role of miR-532-5p in breast cancer cells via direct targeting of RERG expression.

Acute and Subchronic Oral Toxicity Study of Gardenia Yellow E500 in Sprague-Dawley Rats.

OBJECTIVE: This study was conducted to evaluate the acute and subchronic toxicity of gardenia yellow, a natural colorant widely used in China and other Asian countries. An acute toxicity test was performed in S-D rats of both genders and the lethal dose (LD50) of per oral gardenia yellow was estimated to be more than 15.0 g/kg·bw. In the subchronic study, gardenia yellow was orally administered to rats by gavage at doses of 0, 0.50, 1.50 and 4.50 g/kg·bw/day for 90 days followed by a recovery period of 28 days. No appreciable toxic-related changes were observed in the 0.50 g/kg·bw/day group. When the animals received gardenia yellow at 1.50 g/kg·bw/day or more, body weight loss was observed, and pigments began to deposit in several vital organs, resulting in significant changes of several hematological and biochemical indicators related to the nutritional status of the body, liver and kidney function, more severe in the high dose group. In the recovery period, the alterations of the clinical symptoms and parameters were relieved a lot. Based on the results of the current study, the no observed adverse effect level (NOAEL) of gardenia yellow E500 in rats was set to be 0.50 g/kg·bw/day.

Chronic toxicity study in Sprague-Dawley rats on transgenic rice T1c-19 with cry1C* gene.

In the present study, a novel transgenic rice line T1c-19 carrying cry1C* gene was evaluated in Sprague-Dawley (SD) rats by a 52-week feeding study, aiming at determining its unintended effects. The rice T1c-19 and its parental rice were prepared at a level of up to 60.75% in the growth diet and 66.75% in the maintenance diet, respectively. AIN-93 diet was used as a nutritional control. All the diets were nutritionally balanced. Each group, with 48 rats of both genders, was fed the corresponding diet for 52 weeks. The results of clinical signs, body weight and food consumption of the transgenic rice group were comparable to those of the parental rice group. Clinical measurements were made on weeks 13, 26 and 52, and statistical significances were observed in several hematological and serum biochemical indices between the two rice groups and were not considered as treatment-related. The terminal histopathological examination showed some spontaneous lesions in all groups with no significant difference among them. Taken together, the results of the present 52-week chronic toxicity study of transgenic rice T1c-19 exerted no unintended adverse effects on SD rats.

Overexpression of Long Noncoding RNA PTENP1 Inhibits Cell Proliferation and Migration via Suppression of miR-19b in Breast Cancer Cells.

This study aimed to investigate the effect of long noncoding RNA PTENP1 in the development of breast cancer (BC). Quantitative real-time PCR was utilized to determine the expression of PTENP1 in tissues and cell lines. pcDNA3.1 and shRNA were used to over- and low-express PTENP1 in BC cell lines, and miR-19b mimic and inhibitor were utilized to over- and low-express miR-19b. Then the abilities of cell survival, apoptosis, migration, and invasion were assessed in BC cells with different expression levels of PTENP1 and miR-19b. The expression of PTENP1 was significantly downregulated in both BC tissues and cell lines. Overexpressed PTENP1 could significantly increase cell survival, colony forming, migration, and invasion but decrease apoptosis in BC cell lines. However, overexpressed miR-19b performed contrary effects compared with PTENP1 on cell survival, colony forming, migration, invasion, and apoptosis in BC cell lines. miR-19b can be downregulated by PTENP1, and the effect of overexpressed PTENP1 on the PI3k/Akt pathway could be aborted by overexpressed miR-19b. PTENP1 performed a negative role in the development of BC via downregulating miR-19 probably through the PTEN/PI3K/Akt pathway.

Cancer-Associated Fibroblasts Correlate with Tumor-Associated Macrophages Infiltration and Lymphatic Metastasis in Triple Negative Breast Cancer Patients.

Background: Cancer-associated fibroblasts (CAFs) have been shown to be among the most prominent cells in tumor microenvironment and play a significant role in accelerating tumor metastasis by interacting with other type of cells. Tumor-associated macrophages (TAMs), the predominant tumor-infiltrating immune cells, also play important roles in cancer progression. Here, we aimed to evaluate the effects of CAFs on infiltration of TAMs and lymphatic metastasis in triple-negative breast cancer (TNBC). Material and methods: The study included 278 patients with histologically confirmed TNBC. Immunohistochemical staining of α-smooth muscle actin and fibroblast activation protein were used to identify CAFs. Polarized functional status of infiltrated TAMs was detected by expression of CD163. The clinicopathological features were assessed from all the patients' medical records. Results: The CAFs-related markers were found to be expressed more frequently in TNBC patents with aggressive behaviors, including recurrence and poor histological differentiation. High activation of CAFs was positively correlated with elevated infiltration of polarized CD163-positive TAMs and lymph node metastasis in TNBC patients. Multivariate Cox analysis revealed that the activation of CAFs, TAMs infiltration, and lymph node metastasis were independent prognostic factors for disease-free survival in TNBC patients. Conclusion: Cancer-associated fibroblasts were associated with infiltration of CD163-positive macrophages and lymphatic metastasis, and may be potential prognostic predictors of TNBC.

A meta-analysis on the effect of operation modes on the recurrence of papillary thyroid microcarcinoma.

Whether total thyroidectomy reduces the recurrence rate in patients with papillary thyroid microcarcinoma (PTMC) is currently controversy. Conclusions of sporadic, inconsistent, and mono-institutional studies need a meta-analysis to evaluate. 525 relevant studies were obtained from initial search on PubMed, 511 studies were excluded by inclusion and exclusion criteria. Eligible data were extracted from each included study. The Odds ratios (ORs) and 95% confidence interval (CI) were used to assess the difference in the recurrence rates between PTMC patients treated with total thyroidectomy and non-total thyroidectomy. OR and 95% CI were calculated using a fixed-effects or a random-effects model. The Q statistic was used to evaluate homogeneity and Begg's test was used to assess publication bias. 14 studies meeting the inclusion criteria were included in this meta-analysis. The over all recurrence rates of pooled patients with total thyroidectomy and non-total thyroidectomy were 2.83% and 2.84% respectively. Primary random-effects model analysis showed, no significant difference of recurrence rates existed between two operation modes (OR = 0.732, 95% CI: 0.444 - 1.208), while, high heterogeneity among studies was found, I-squared index (I2) = 40.2%. After remove one study with high heterogeneity, the OR of the pooled recurrence rates of the total thyroidectomy and the non-total thyroidectomy groups was 0.786 (95% CI: 0.363 - 1.701), further suggesting no significant difference of the recurrence rate exists between two operation modes. Our meta-analysis demonstrated postoperative recurrence of PTMC is not reduced by total thyroidectomy, non-total thyroidectomy is also a good choice to treat PTMC patients.

IgG Expression upon Oral Sensitization in Association with Maternal Exposure to Ovalbumin.

The role of maternal allergen exposure in the allergenicity of the offspring remains controversial. Some studies have shown that maternal exposure is a risk factor for allergy in the offspring, whereas other studies have shown that maternal exposure induces immune tolerance and protects offspring from allergy disease. Therefore, we utilized maternal rat allergen exposure model to evaluate the offspring immune reactions to ovalbumin protein and to determine whether the Brown Norway (BN) rat model is a suitable animal model for studying the allergenicity of food proteins. For three generations, rats received an allergens or non-allergens by gavage during the pregnancy and lactation periods. After weaning, the offspring rats were used for oral sensitization experiment. In the sensitization experiment, the control rat, which had maternal exposure to phosphate-buffered saline (PBS), exhibited full response of IgG to oral exposure to OVA. The IgG level was significantly lower in F1 rats that were sensitized by maternal exposure to ovalbumin(OVA). Moreover, the lowest IgG level was found for the F3b sensitized by maternal rats exposed to OVA allergen for three continuous generations. Compared with maternal OVA exposure prior to postnatal sensitization, the sensitization via maternal PBS led to a higher serum level of OVA-specific IgG. However, the OVA-specific IgG levels for the two generations of maternal PBS exposure prior to postnatal sensitization was not higher than that for the one generation of maternal rats exposed to PBS prior to postnatal sensitization. Our studies demonstrate that maternal OVA exposure during the pregnancy and lactation can affect the results of oral sensitization studies using ovalbumin protein. BN rats must be bred in non-allergen conditions for at least one generation to avoid problems in rat models for studying the allergenicity of food proteins.

The associations between mast cell infiltration, clinical features and molecular types of invasive breast cancer.

Associations between mast cell infiltration and the clinical features and known molecular profile of breast cancer remain unclear. The distribution difference of mast cell was evaluated, in 219 patients with no special type of invasive carcinoma, using sorts of age, max diameter of cancer, histological type, lymph node metastasis as well as the expressions of estrogen receptor (ER), progestogen receptor (PR), human epidermal growth factor receptor 2 (HER-2) and nuclear protein Ki67. The mast cell density (MCD) in patients younger than 50 years old was significantly higher than that in patients with age ≥ 50. The MCD in ER or PR positive patients was significantly higher than MCD in ER or PR negative patients. The MCD in patients with Ki67 ≤ 14% was also significantly higher than MDC in patients with Ki67 > 14%. The MCD of patients with invasive ductal carcinoma was significantly higher than MCD of patients with invasive lobular carcinoma. No significant distribution difference of MCD was found to be associated with max diameter of cancer, lymph node metastasis and HER-2. Further analysis found that MDC was significantly higher in patients after neo-adjuvant chemotherapy. The distribution difference of mast cell widely exists in patients with distinct clinical features, the role of mast cell in breast cancer need further research with detailed and reasonable classification to clarify.

[Enhanced absorption of breviscapine photosomes in small intestine of rats].

OBJECTIVE: To investigate whether the absorption of breviscapine can be enhanced by using breviscapine photosomes. METHOD: Testing the uptake and intestinal permeability of breviscapine powder and breviscapine photosomes by using intestine perfusion technique and reverted gut sac method. RESULT: The uptake and permeability coefficient of breviscapine were increased in photosomes. The absorption process obeyed the Fick's law in the range of 0-100 microg x mL(-1). CONCLUSION: The absorption breviscapine photosomes is enhanced by increasing the permeability through a passive mechanism.

[Study on effects of Erigeron breviscapus extract on anticoagulation].

OBJECTIVE: To study the effect of Erigeron breviscapus extract on anticoagulation. METHOD: Coagulation time (CT), prothrombin time (PT), platelet factor III (PF3) and euglobulin lysis time (ELT) have been used to evaluate the anticoagulation activities of Erigeron breviscapus extract. RESULT: Erigeron breviscapus extract significantly delayed CT and PT, inhibited the activity of PF3, and decreased ELT. CONCLUSION: Erigeron breviscapus extract plays an role in anticoagulation by inhibiting PF3 and prothrombin V. It can also enhance the activity of fibrimolysis.