Five Constituents Contributed to the Psoraleae Fructus-Induced Hepatotoxicity via Mitochondrial Dysfunction and Apoptosis

Backgrounds: Psoraleae Fructus (PF)-induced hepatotoxicity has been reported in clinical and animal experiments. However, the hepatotoxic constituents and mechanisms underlying PF-induced toxicity have remained unclear. Therefore, this study explored the potentially toxic PF components and revealed their relative mechanisms.Methods: The hepatotoxicity of PF water (PFW) and ethanol (PFE) extracts was compared using Kunming mice. The different compositions between PFW and PFE, which were considered toxic compositions, were identified using the UHPLC-Q-Exactive MS method. Then, L02 and HepG2 cell lines were used to evaluate the toxicity of these compositions. Cell viability and apoptosis were determined through the Cell Counting Kit-8 (CCK-8) assay and flow cytometry, respectively. An automatic biochemical analyzer detected the aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP). Lastly, we used high-content screening (HCS) to determine the levels of reactive oxygen species (ROS), lipid, and mitochondrial membrane potential (MMP).Results: The ethanol extraction process aggravated the hepatotoxicity of PF, causing more severe injuries. The content of psoralen, isopsoralen, bavachin, psoralidin, bavachinin, neobavaisoflavone, and bakuchiol was higher in the PFE than PFW. Bavachin, psoralidin, bavachinin, neobavaisoflavone, and bakuchiol induced cell apoptosis and the AST, ALT, and ALP leakages. Furthermore, these five constituents increased intracellular lipid accumulation and ROS levels but decreased the MMP level.Conclusion: The ethanol extraction process could induce severe PF hepatotoxicity. Bavachin, psoralidin, bavachinin, neobavaisoflavone, and bakuchiol are the main hepatotoxic ingredients. This mechanism could be associated with oxidative stress and mitochondrial damage-mediated apoptosis. Taken together, this study provides a basis for the clinical application of PF that formulates and improves its herbal standards.

PSVII-22 Changes in biochemical composition of saliva and blood of cattle after sunflower husk included in the diet

The aim of this study was to assess the effect of the introduction of food waste sunflower husk (SH) into the diet on the change in the chemical composition of biological media (blood and saliva) against the background of control (C). The studies were carried out on young 8-month cattle; ration of C - 70% roughage, 25% - concentrated feed, 5% - premix; SH - ration C + 10% sunflower husk (shredded, 0.5–2 mm), instead of the coarse part of the ration. The selection of biosubstrates (saliva, blood) was carried out in the morning before feeding: saliva was obtained by the method of local pharmacological stimulation of salivation (2% sodium citrate solution), blood from the jugular vein into vacuum tubes with a coagulation activator. The chemical composition of biological media was carried out on an automatic biochemical analyzer CS-T240 (Dirui Industrial Co., Ltd., China) using commercial biochemical kits for veterinary medicine (DIAKON-DS, Russia; Randox Laboratories Ltd, Great Britain). The inclusion of SH in the diet contributed to an increase in serum urea by 82.4% (P ≤ 0.05), lipase enzyme by 81.5% (P ≤ 0.05), uric acid by 62.7% (P ≤ 0.05), Ca and P by 73% and 21.8% (P ≤ 0.05), while a decrease in p-Amylase by 63.6% (P ≤ 0.05) and Fe by 86.8% (P ≤ 0.05) relative to C. After SH administration an increase in urea by 73% (P ≤ 0.05), digestive enzymes - lipase by 76.4% (P ≤ 0.05), p-amylase by 61 % (P ≤ 0.05), calcium by 69.3% and iron by 45% (P ≤ 0.05) and superoxide dismutase enzyme in comparison with C was registered in saliva. The obtained data show an increase in the level of digestive enzymes and elements in blood and saliva of ruminants, which favorably contributes to the use of sunflower husk in feeding cattle. This research was performed with financial support from the RSF (# 20-16-00088).

Evaluation of Colocasia esculenta Schott in anti-cancerous properties with proximity extension assays

Background: Colocasia esculenta Schott (called as Xiangshayu in Chinese) is an excellent local cultivar of the genus polymorpha in Jiangsu Province, China. Objective: In the present study, we have performed a comparative study before and after dietary consumption with Colocasia esculenta Schott to evaluate its anti-cancerous properties. Design: Forty-two healthy volunteers were recruited, and dietary consumption with 200 g of tap water cooked Colocasia esculenta Schott daily was conducted for 1 month. Plasma samples from the subjects before and after dietary consumption with Colocasia esculenta Schott were analyzed with proximity extension assays for the alteration of 92 proteins in relation with cancers, while blood samples were examined for physiological parameters with an automatic biochemical analyzer. Bioinformatic analyses were conducted using MalaCards and GEPIA. Results: After taking dietary consumption with Colocasia esculenta Schott, circulating CYR61, ANXA1, and VIM protein levels in the subjects was found to be most significantly downregulated, while for ITGB5, EPHA2, and CEACAM1, it was upregulated. Alternation of these proteins was predicted to be associated with the development of tumors such as pancreatic adenocarcinoma and breast and prostate cancers. Conclusion: The present study provides evidence that Colocasia esculenta Schott, as a healthy food, has anti-cancerous properties. Further investigation of phytochemistry in Colocasia esculenta Schott has been taken into our consideration.

The effect of photodynamic therapy on the hematological and biochemical parameters of the blood of cats

Relevance. In this article, we present experimental study on the effect of photodynamic therapy with a chlorin-type photosensitizer on the hematological and biochemical blood parameters of cats with malignant oncological diseases.Methods. During conducting studies in experimental animals, we took blood samples from 44 cats aged 8 to 16 years, of different breeds and different sexes, before photodynamic therapy and two days after treatment, in order to determine the possible effect of therapy on the blood counts of patients. Blood sampling was carried out according to the standard method. Hematological examination was performed on a PCE90vet analyzer (HTI, USA), biochemical on an automatic biochemical analyzer Biosystems A15 (Biosystems, Spain) and on a semi-automatic analyzer Biohaem SA (HTI, USA). Photodynamic therapy was performed according to the standard method, with the preliminary introduction of the photosensitizer “Photoditazine” at a dose of 0.8– 1 mg/kg, 3 hours before irradiation.Results. The most significant changes in the hematological study were found among the following parameters: the level of white blood cells after irradiation increased by 18%, the number of eosinophils decreased by 28%, the number of segmented neutrophils increased by 11%, the content of lymphocytes decreased by 21%. When studying the biochemical parameters of blood, changes were found in the following parameter: the glucose level increased by 13% after irradiation, the level of GGT decreased by 19% after irradiation. At the same time, all indicators of the hematological and biochemical composition of the blood were within the normal values for this type of animal. Photodynamic therapy does not significantly affect the hematological and biochemical parameters of the blood of cats, this proves the safety of using this method of treatment in these animals.

Up-regulated YKL-40 is associated with poor prognosis of HCC patients with hepatitis B related cirrhosis

IntroductionTo investigate clinical significance of YKL-40 in hepatocellular carcinoma (HCC) patients with hepatitis B (HBV) related cirrhosis.Material and methodsThe present prospective observational study included 129 cases of HCC patients. Besides, 152 patients with only hepatitis B related cirrhosis and 110 HCC patients with no cirrhosis were also enrolled during the same period. Additionally, 100 healthy individuals were enrolled as control. Serum YKL-40 levels were determined using ELISA method. Levels of serum albumin, total bilirubin, alanine aminotransferase (ALT) and aspartate transaminase (AST) as well as HCC related biomarkers of alpha fetoprotein (AFP), des-γ-carboxy prothrombin (DCP), gamma glutamyltransferase (GCT-II), α-L-Fucosidase (AFU), CEA and CA19-9 were measured using an automatic biochemical analyzer. Patients’ demographic and clinical characteristics were collected and analyzed.ResultsThe expression of YKL-40 was the highest in HCC patients with HBV-related cirrhosis and the lowest in the healthy control, and the difference was significant compared with other groups. And HCC patients showed markedly higher YKL-40 levels than the HBV-related cirrhosis patients. Patients with higher expression of YKL-40 showed higher ratio of TNM IV, lymphatic metastasis and Child-Pugh C, and higher serum levels of AFP, AFU and CA19-9 than those in the patients with lower levels of YKL-40. YKL-40 level was positively correlated with AFP and AFU. Survival analysis showed patients with higher expression of YKL-40 had shorter 1-year survival time than the patients with lower YKL-40.ConclusionsYKL-40 was elevated in HCC patients with HBV-related cirrhosis and high expression of YKL-40 predicted poor prognosis and shorter 1-year survival.

Association of ApaI polymorphism of VDR gene with obesity in Iranian population

Introduction: Identifying the risk factors of obesity as a problem facing communities is crucial given its complexity. The vitamin D receptor (VDR) gene has been reported as a candidate for this disease in literature. Objective: To investigate the association of ApaI, BsmI and TaqI polymorphisms of VDR gene with obesity in part of Iranian population. Material and methods: In this study, genotypes of 348 obese (BMI ≥30 kg/m2) and 320 non-obese (BMI: 18.5-24.9 kg/m2) were analyzed by using PCR-RFLP method. The levels of FBS, TG, Total Cholesterol, HDL- Cholesterol and LDL- Cholesterol were measured by using an automatic biochemical analyzer. Results: The present findings showed significantly higher BMI, FBS and TG in the obese group compared to in the controls. In the obese individuals, the frequency of genotype AA was 47.1% and that of combined genotype Aa+aa 52.9%, whereas these figures were respectively 30% and 70% in the controls [P=0.024, 95% confidence interval (CI)=1.100-3.933, odds ratio (OR)=2.08]. The frequencies of A and a alleles for the ApaI polymorphism were statistically significant in the two groups (allele A vs. a, P=0.017). No significant relationships were also observed between TaqI genotypes and alleles in the controls and obese subjects. Conclusion: The present study found VDR ApaI (rs7975232 C/A) polymorphism appeared to be a risk factor for obesity. In particular, A allele and genotype of AA in ApaI were associated with the obesity phenotypes.

Gualou Xiebai Banxia decoction ameliorates Poloxamer 407-induced hyperlipidemia

Ethnopharmacological relevance: Gualou Xiebai Banxia decoction (GLXBBX) is a well-known traditional Chinese herbal formula that was first discussed in the Synopsis of the Golden Chamber by Zhang Zhongjing in the Eastern Han Dynasty. In traditional Chinese medicine, GLXBBX is commonly prescribed to treat cardiovascular diseases, such as coronary heart disease and atherosclerosis. Objective: The present study aimed to examine GLXBBX’s preventative capacity and elucidate the potential molecular mechanism of Poloxamer 407 (P407)-induced hyperlipidemia in rats. Materials and Methods: Both the control and model groups received pure water, and the test group also received a GLXBBX decoction. For each administration, 3 mL of the solution was administered orally. To establish hyperlipidemia, a solution mixed with 0.25 g/kg P407 dissolved in 0.9% normal saline was injected slowly into the abdominal cavity. At the end of the study, the rats’ plasma lipid levels were calculated using an automatic biochemical analyzer to evaluate the preventative capability of the GLXBBX decoction, and the serum and liver of the rats were collected. Results: The GLXBBX decoction significantly improved P407-induced hyperlipidemia, including increased plasma triglycerides, aspartate aminotransferase elevation, and lipid accumulation. Moreover, GLXBBX decoction treatment increased lipoprotein lipase (LPL) activity and mRNA expression of LPL. Furthermore, GLXBBX significantly suppressed the mRNA expression of stearoyl-CoA desaturase (SCD1). Conclusion: GLXBBX significantly improved P407-induced hyperlipidemia, which may have been related to enhanced LPL activity, increased LPL mRNA expression, and decreased mRNA expression of SCD1.

Validation of the biochemical method for determining the total cholesterol level in the samples of biological fluids as a base for providing the quality of dyslipidemia diagnostics

The main principles of creating a quality system in modern laboratory diagnostics are: standardization of laboratory processes by developing standard operating procedures; general quality management of laboratory research based on the development and implementation of the requirements of international standards (according to ISO 15189: 2015 "Medical laboratories. Basic requirements for quality and competence"); quality control of all stages of the laboratory process through the implementation of the validation procedure. Aim. To develop a methodology for conducting validation procedures to assess the suitability of a biochemical method for determining the level of total cholesterol in biological fluids in the Clinical Diagnostic Laboratory of the Clinical Diagnostic Center of the NUPh. Materials and methods. The object of the study was a standardized method for determining the concentration of total cholesterol. The method was validated using the “Cholesterol Reagent Set” test kit and the standard sample “Chemical control. Reagent kit. Level 1 ", manufactured by High Technology, Inc. (USA) with known concentration. The measurements were carried out on an Express Plus automatic biochemical analyzer manufactured by Bayer Corporation, Germany. When processing the research results, descriptive statistics were used and a number of statistical evaluations were carried out. Results. A protocol and a validation report were developed at the Clinical Diagnostic Laboratory of the CDC NUPh to assess the suitability of the method for determining the concentration of cholesterol in biological fluids by the photometric enzymatic method on an automatic biochemical analyzer Express Plus (using reagents and control material manufactured by High Technology, Inc., USA). The validation characteristics of the method were determined: repeatability and reproducibility, correctness and uncertainty of measurements. Evaluation of the internal laboratory repeatability and reproducibility of this technique indicates the absence of gross errors in the operation of the instrument and statistically important differences in measurements. The assessment of the correctness of the method (carried out using the control material) proved that the systematic error is not significant (according to a given acceptance criterion). The expanded uncertainty calculation showed that the obtained values ​​of the total cholesterol level can be considered accurate and reliable. Conclusions. Validation of the method for determining total cholesterol in human blood by the photometric enzymatic method has proven that this method has performance characteristics that correspond to the regulated ones, meets the established criteria, and the parameters measured with it correspond to the proper ones. Key words: validation, determination method, total cholesterol, repeatability and reproducibility, correctness and uncertainty of measurements

Liver metabolic activities of Pasundan cattle induced by irradiated chitosan

Mushawwir A, Arifin J, Darwis D, Puspitasari T, Pengerteni DS, Nuryanthi N, Perman R. 2020. Liver metabolic activities of Pasundan cattle induced by irradiated chitosan. Biodiversitas 21: 5571-5578. A total of one hundred and twenty-five, 2-3 year old male Pasundan cattle were used as livestock samples during the three months of this research. They were selected from the local cattle breeding and development center in Ciamis. The animal samples were randomly allocated to 5 treatment groups. One group served as the control, or without irradiated chitosan, while the others were used as treatment in varying levels. Each treatment group involved five replicates with 25 Pasundan bulls per treatment i.e five Pasundan bulls per replication. Each group was provided with the following rations: C0 = Control group, without IC (0 ppm IC); C1 = 350 ppm Irradiated Chitosan (IC); C2 = 400 ppm IC; C3 = 450 ppm IC; and C4 = 500 ppm IC. Irradiated chitosan was obtained through the following steps: extraction, deacetylation, and irradiation of chitin using gamma rays. Five mL of blood samples were collected from each bull at the beginning of each month of this experiment, which totaled three months. The blood samples were sucked from the tail/coccygeal vein using a sterilized syringe and vacuum tube containing K3EDTA. The plasma was used to determine the concentration of parameters related to liver metabolism through an automatic biochemical analyzer Kenza 240TX model from Biolabo, using a commercial kit. Each procedure was followed based on the Biolabo kit (Franch) and Randox kit (UK). This study showed that IC reduces the activity of glycogenolysis and glycolysis, but is accompanied by improvements in the biochemical conditions of liver cells. This is a favorable condition for the metabolism of Pasundan bulls in order to enhance their growth and reproduction.

Hematological and antioxidants responses of dairy cow fed with a combination of feed and duckweed (Lemna minor) as a mixture for improving milk biosynthesis

Tanuwiria UH, Mushawwir A. 2020. Hematological and antioxidants responses of dairy cows fed with a combination of feed and duckweed (Lemna minor) as a mixture for improving performance. Biodiversitas 21: 4741-4746. A total of twenty-five, 5-7th-lactation-old or 7-8thyears-old dairy cows were used in the current study to study the effect of feed duckweed (Lemna minor) on the hematological status and parameters related antioxidant in the dairy cow. Each group of treatment involved 5 replicates with a dairy cow each (5 dairy cows per group). All of the group was provided ration following F0 = Uncultivated grass (UG) 60% and supplemented concentrated (C) 40%; F1 = UG 50%+fresh duckweed 10%+C 40%; F2 = UG 60%+fresh duckweed 3%+C 37%; F3 = UG 50%+fresh duckweed 10%+dried duckweed 3%+C 37%; F4 = UG 45%+fresh duckweed 15%+dried duckweed 2%+C 38%. Blood samples were collected from each dairy cow monthly beginning at the first month of this experiment, from the tail vein (vena coccygeal), using a sterilized syringe and vacuum tube containing K3EDTA. Hematological parameters were analyzed by a hematology analyzer. The blood samples collected were also centrifuged to separate the plasma. The plasma was used to determine of concentration of parameters related to antioxidant by an automatic biochemical analyzer, using a commercial kit. All procedure of the analysis was following based on the Biolabo kit (Franch) and Randox kit (UK). Based on in this current study showed that increased hematological condition and antioxidants profile in dairy cow supplemented duckweed . Supplemented duckweed can be improved physiological condition (hematologic and antioxidants) in the dairy cow.