Currently, the presence of antimalarial drug resistance has become a major obstacle in the treatment of malaria. To overcome the problem, a series of studies are needed to find new antimalarial drugs from plants. Previously, 90% ethanolic extract of Cassia spectabilis DC (EECS) leaves have been reported to have antimalarial activity in vitro against Plasmodium falciparum and in vivo against Plasmodium berghei ANKA. The research is conducted to find out the toxicity and protective effects of EECS on the liver and kidneys of mice infected with P. berghei ANKA. The acute and subacute toxicity tests were carried out on healthy mice that were given EECS at a dose of 150 mg/kg BW. An antimalarial activity test was carried out at doses of 150 and 200 mg/kg BW in P. berghei-infected mice. Regarding hepatomegaly, further plasma levels of hepatic enzyme were analyzed, as well as histopathological observation of the liver to determine the effect of the extract on liver. The kidney was observed histopathologically as well. The acute toxicity test of EECS showed that there was no mouse died at the highest dose, indicating safe for the mice. The subacute toxicity based on the histology data showed no significant difference in the liver and kidney of mice between the tested group and the healthy group. The histological and enzymatic effect of EECS in mice infected with P. berghei showed the histological and enzymatic effect that improved liver function and the histopathological effect on kidneys with the highest activity at a dose of 200 mg/kg BW compared with the negative control. The results showed the EECS was not toxic in mice and repaired the liver and kidney functions of P. berghei ANKA-infected mice, indicating a good candidate for antimalarial drug development.

This study was aimed to evaluate the metabolite contents and antineuroinflammatory potential of Marsilea crenata Presl. grown under a controlled environmental condition. The antineuroinflammatory test has been carried out in vitro using ethanolic extract of M. crenata leaves on HMC3 microglia cells. An in silico approach was applied to predict the active compounds of the extract. The HMC3 microglia cells were induced with IFNγ to create prolonged inflammatory conditions and then treated with 96% ethanolic extract of the M. crenata leaves of 62.5, 125, and 250 μg/mL. The expression of MHC II was analyzed using the ICC method with the CLSM instrument. Metabolites of the extract were profiled using UPLC-QToF-MS/MS instrument and MassLynx 4.1 software. In silico evaluation was conducted with molecular docking on 3OLS protein using PyRx 0.8 software, and physicochemical properties of the compounds were analyzed using SwissADME webtool. The ethanolic extract of M. crenata leaves could reduce the MHC II expression in HMC3 microglia cells in all concentrations with the values 97.458, 139.574, and 82.128 AU. The result of metabolite profiling found 79 compounds in the extract. In silico evaluation showed that 19 compounds gave agonist interaction toward 3OLS, and three met all parameters of physicochemical analysis. The ethanolic extract of the environmental-controlled growth of M. crenata leaves antineuroinflammatory activity on HMC3 microglia cells. The extract was predicted to contain some phytoestrogen compounds which act as 3OLS agonists.

Estrogen deficiency can contribute to osteoporosis in postmenopausal women. Phytoestrogens are becoming more widely recognized as potential estrogen replacement therapy. The administration of phytoestrogens can cause bone formation, which is marked by an increase in Runx2 expression in osteoblast cells and can be seen using western blot and immunohistochemistry approaches. This review aimed to compare the detection methods of Runx2 in phytoestrogen-induced bone tissue using western blots and immunohistochemistry. Selectivity, sensitivity, processing time, and cost-effectiveness were the parameters that were compared. This review was done by identifying articles in several databases (Google Scholar, PubMed, and Science Direct). The process of selecting the articles used the PRISMA guidelines to create a flowchart with inclusion and exclusion study criteria. Meta-synthesis was done to analyze, identify, and interpret all of the data in the articles systematically. 70 articles in total were obtained from the selection process, with 21 articles being relevant to the topic. The result shows that the selectivity and sensitivity of western blot for detecting Runx2 on tissue were 93.5–100%, respectively, whereas immunohistochemistry selectivity and sensitivity were 45–99.5%, respectively. Compared to immunohistochemistry, western blot can save up to 57.26%. Immunohistochemistry takes 46 hours to process, while Western blot takes 25 hours and 20 minutes. In comparison to immunohistochemistry, the western blot is more selective, sensitive, rapid and affordable for detecting Runx2 in bone tissue.

Osteoporosis is a bone disorder characterized by the decrease of bone mass along with bone micro-architecture damage and has a risk become a fracture. One of the causes of osteoporosis is estrogen deficiency. Genistein is a phytoestrogen compound in the isoflavone group containing a similar structure compared to 17β-estradiol, thus it can bind to estrogen receptors and produce an estrogenic effect. Genistein induction can stimulate bone formation and promote the increase of alkaline phosphate (ALP) activities in osteoblast cells which can be observed by immunocytochemistry or Enzyme-linked Immunosorbent Assay (ELISA) or Western blot method. Using the PRISMA guideline technique, choose and strategize article searches by reading the title, abstract, and then the whole text of the article. Articles with the keywords "genistein or osteoblast cells or alkaline phosphate or immunocytochemistry or immunofluorescence or ELISA or western blot" were retrieved from databases including Google Scholar, PubMed, Researchgate, and Sciencedirect. 24 relevant research articles were uncovered as a result of this systematic review. Comparison of immunocytochemistry and ELISA methods in order to analyze the activities of ALP in osteoblast induced by genistein includes selectivity, sensitivity, processing time, and cost efficiency parameters. The immunocytochemistry method has a higher level of sensitivity and a faster processing time, whereas the ELISA method has a higher level of selectivity and less cost efficiency. The western blot method has selectivity for detecting complex-level protein expression.