Differential protein expression of the inhibitory effects of a standardized extract from Scutellariae radix in liver cancer cell lines using liquid chromatography and tandem mass spectrometry

The purpose of this study is to verify the inhibitory effect of a chemically standardized extract from Scutellariae radix in liver cancer cell lines (HepG2). The botanical extract was prepared using pressurized liquid extraction (PLE). A method using proteolytic digest with single dimensional and two-dimensional liquid chromatography with tandem mass spectrometry was used to characterize differential protein expression in mammalian cells in response to the botanical extract. The whole cell lysates were digested with trypsin, and the peptides were separated by one-dimensional (reversed phase) or by two-dimensional (cation exchange and reversed phase) solid-phase extraction (SPE) cleanup and separated by liquid chromatography with UV detection and mass spectrometry. In the presence of the botanical extracts, drug-induced apoptosis was not observed, and a number of proteins that played an important role in the metabolic pathways in HepG2 cell line had been affected. The data, as presented, suggest that the inhibitory effects of the standardized extracts from Scutellariae radix resulted from expression of heat shock protein and other proteins related to energy metabolism. The proposed platform had the potential to provide significant information about the particular proteome such as human hepatoma HepG2. At the molecular level, it was possible to study the proteins and how their levels and modifications change in response to the effects of the botanical extract.     

Ciprofloxacin and ofloxacin aqueous humor concentrations after topical administration in dogs undergoing cataract surgery

OBJECTIVE: To determine if preoperative topical administration of ciprofloxacin or ofloxacin results in aqueous humor drug concentrations that exceed the MIC(90) of common ocular contaminants in dogs undergoing cataract surgery. PROCEDURES: Twelve dogs were treated with topical 0.3% ciprofloxacin and 13 dogs with topical 0.3% ofloxacin once the night before surgery, and then with 1 drop of ciprofloxacin or ofloxacin every 15 min for 2 h immediately before surgery. Aqueous humor samples were taken from each eye immediately before the incision was made and frozen at -70 degrees C. First eye samples (S1) were taken closer to the time of topical treatments than second eye samples (S2). Samples were analyzed by high performance liquid chromatography (HPLC) at the North Carolina State University (NCSU) Clinical Pharmacology laboratory. RESULTS: In ciprofloxacin patients, S1 concentrations were 0.03-0.69 (0.17 median) microg/mL, and S2 concentrations were 0.09-0.95 (0.36 median) microg/mL. Aqueous humor concentrations did not exceed the MIC90 of Streptococcus sp. Few eyes (1/12 OU) exceeded the MIC90 for Staphylococcus sp. or Corynebacterium sp.; moderate numbers (5/12 S1; 8/12 S2) exceeded the MIC90 for E. coli. In ofloxacin patients, S1 concentrations were 0.48-2.81 (1.05 median) microg/mL, and S2 concentrations were 0.45-3.63 (1.30 median) microg/mL. Although few eyes (相似文献   

Intraepidermal adenocarcinoma in the perianal skin of two cats, a condition resembling human extramammary Paget's disease

In humans, mammary and extramammary Paget's disease is an uncommon to rare manifestation of intraepidermal adenocarcinoma arising from simple epithelium, usually glandular in origin. This report describes two cats with lesions in perianal skin consisting of atypical intraepidermal neoplastic cells. Differential diagnoses included intraepidermal adenocarcinoma, in situ squamous or basal cell carcinoma, junctional amelanotic melanoma, and epitheliotropic tumours of histiocytic or lymphocytic origin. The atypical intraepidermal cells in the cats were immunohistochemically positive for cytokeratin 8/18 (CK8/18), which stains simple (glandular) epithelium. The keratinocytes and basal cells were negative for CK8/18. In addition, the atypical intraepidermal cells were immunohistochemically negative for melanocytic, lymphocytic, and histiocytic markers. The staining results confirmed the atypical intraepidermal cells to be of simple glandular origin, and ruled out other causes of intraepidermal malignancy. In one cat the clinical lesions consisted of a pruritic erythematous eruption surrounding the anus. Another cat presented clinically for an area of irregular anal thickening; this cat had well-regulated diabetes mellitus. The cats were otherwise clinically healthy. The clinical features, histological appearance, and immunohistochemical staining of the skin lesions were consistent with those described for human perianal extramammary Paget's disease. To the authors' knowledge, this is the first report of an intraepidermal adenocarcinoma in a cat or other animal species.     

Intrahepatic portosystemic venous shunt in a neonatal roan antelope (Hippotragus equinus cottoni).

A 6-day-old, female roan antelope (Hippotragus equinus cottoni) was diagnosed with a single intrahepatic portosystemic venous shunt at necropsy. Clinical signs had included weakness, lethargy, hypothermia, diarrhea, and a weak suckle response. Multiple seizure episodes were associated with hypoglycemia and characterized by vocalization, muscle fasciculations, and disorientation. Hematologic abnormalities included anemia with hypochromasia, anisocytosis, poikilocytosis, and leukopenia with neutropenia and lymphopenia. Serum biochemical abnormalities included elevations in blood urea nitrogen and total serum bile acid concentration. A portosystemic vascular anomaly should be a differential diagnosis for nonthriving, exotic ruminant calves with overt or subtle neurologic signs, persistent hypoglycemia, and/or elevated bile acids. In very young calves, total bile acid concentration may be more useful in establishing a diagnosis than blood ammonia concentration.     

Exposure to the Neurotoxic Dinoflagellate,Alexandrium catenella,Induces Apoptosis of the Hemocytes of the Oyster,Crassostrea gigas

This study assessed the apoptotic process occurring in the hemocytes of the Pacific oyster, Crassostrea gigas, exposed to Alexandrium catenella, a paralytic shellfish toxins (PSTs) producer. Oysters were experimentally exposed during 48 h to the toxic algae. PSTs accumulation, the expression of 12 key apoptotic-related genes, as well as the variation of the number of hemocytes in apoptosis was measured at time intervals during the experiment. Results show a significant increase of the number of hemocytes in apoptosis after 29 h of exposure. Two pro-apoptotic genes (Bax and Bax-like) implicated in the mitochondrial pathway were significantly upregulated at 21 h followed by the overexpression of two caspase executor genes (caspase-3 and caspase-7) at 29 h, suggesting that the intrinsic pathway was activated. No modulation of the expression of genes implicated in the cell signaling Fas-Associated protein with Death Domain (FADD) and initiation-phase (caspase-2) was observed, suggesting that only the extrinsic pathway was not activated. Moreover, the clear time-dependent upregulation of five (Bcl2, BI-1, IAP1, IAP7B and Hsp70) inhibitors of apoptosis-related genes associated with the return to the initial number of hemocytes in apoptosis at 48 h of exposure suggests the involvement of strong regulatory mechanisms of apoptosis occurring in the hemocytes of the Pacific oyster.     

Comparison of dietary variety and ethnic food consumption among Chinese,Chinese-American,and white American women

The study's purpose was to estimate the variety of foods consumed within standard and ethnic food categories by three groups of women between 18 and 35 years of age. Foreign-born Chinese women [N = 21], Chinese-American women [N = 20] and white American women [N = 23] kept 4-day food records, after instruction. Analysis of variance showed that the mean number of different foods consumed by the foreign-born Chinese was significantly [p < 0.05] lower than those eaten by the other two groups for breads/cereals, dairy products, ethnic foods, fats, meats, sweets/sugars, and vegetable categories. White American women consumed significantly more dairy products and legumes/nuts than either of the groups of Chinese women. Diets of the Chinese-American women were more nutrient dense than those of the women in the other groups, containing significantly more energy, riboflavin, iron, folacin, and calcium than those of the foreign-born Chinese women. The percentage of kilocalories derived from fat [33–34%] did not differ among groups. 95% of foreign-born and 85% of Chinese-American women affirmed their ethnic identification by consuming foods belonging to Chinese cuisine; whereas, the percentage [30%] of white American women selecting Chinese food was significantly lower. Mexican and Italian foods were selected by significantly more white and Chinese-American women than by foreign-born Chinese women. Approximately 20% of both groups of Chinese women, but 49% of white American subjects, ate Japanese food. Chinese-American women retained a preference for Chinese food, similar to that of the foreign-born Chinese women. However, the Chinese-American women consumed cheeses, legumes, raw vegetables in salads, and foods from Italian and Mexican cuisines, indicating their acceptance of foods commonly consumed by white American women. Dietary acculturation among the Chinese-American women improved their diets over those of the foreign-born Chinese women and these results support the theory that consumption of a greater variety of foods increases nutrient density of diets.     

Clinical pharmacokinetics and outcomes of oral fluconazole therapy in dogs and cats with naturally occurring fungal disease

This multi-institutional study was designed to determine the clinical pharmacokinetics of fluconazole and outcomes in client-owned dogs (n = 37) and cats (n = 35) with fungal disease. Fluconazole serum concentrations were measured. Pharmacokinetic analysis was limited to animals at steady state (≥72 hr of treatment). The mean (range) body weight in 31 dogs was 25.6 (2.8–58.2) kg and in 31 cats was 3.9 (2.4–6.1) kg included in pharmacokinetic analyses. The dose, average steady-state serum concentrations (C_SS), and oral clearance in dogs were 14.2 (4.5–21.3) mg/kg/d, 26.8 (3.8–61.5) µg/mL, and 0.63 ml min⁻¹ kg⁻¹, respectively, and in cats were 18.6 (8.2–40.0) mg/kg/d, 32.1 (1.9–103.5) µg/mL, and 0.61 ml min⁻¹ kg⁻¹, respectively. Random inter-animal pharmacokinetic variability was high in both species. Two dogs had near twofold increases in serum fluconazole when generic formulations were changed, suggesting lack of bioequivalence. Median C_SS for dogs and cats achieving clinical remission was 19.4 and 35.8 µg/ml, respectively. Starting oral doses of 10 mg/kg q12h in dogs and 50–100 mg total daily dose in cats are recommended to achieve median C_SS associated with clinical remission. Due to the large pharmacokinetic variability, individualized dose adjustments based on C_SS (therapeutic drug monitoring) and treatment failure should be considered.