Iranian Journal of Basic Medical Sciences
Volume:23 Issue: 10, Oct 2020

The present systematic review and meta-analysis was conducted to investigate the effects of soy intake on liver enzymes, lipid profile, anthropometry indices, and oxidative stress in non-alcoholic fatty liver disease (NAFLD). A systematic search was undertaken in PubMed, Embase, Scopus, Web of Science, and Cochrane Library covering up to 10 January 2020. A fixed-effect or random-effects models were applied to pool mean difference (MD) and its 95 % confidence intervals (CI). Four clinical trials comprising 234 participants were included in the meta-analysis. Compared to the controls, alanine aminotransferase (ALT) levels (MD=-7.53, 95% CI=[-11.98, -3.08], P=0.001, I2=0.0 %), body weight (MD=-0.77, 95 % CI=[-1.38, -0.16], P=0.01, I2=36.9%), and the concentration of serum Malondialdehyde (MDA) (MD=-0.75, 95% CI=[-1.29, -0.21], P=0.007, I2=63.6%) were significantly changed following soy intake. Lipid profile was not significantly affected by soy intake. Moreover, no evidence of a significant publication bias was found. The present study suggests lowering effects for soy intake on ALT levels, body weight, and MDA in nonalcoholic liver patients. Therefore, further large-scale and well-designed clinical trials are needed to find conclusive findings.

Methicillin-resistant Staphylococcus aureus (MRSA) is one of the major health hazards and became of greater public health concern since the emergence of community-acquired MRSA. This work aimed to study the prevalence of mecA, femA, femB, lukS-PV, lukF-PV (PVL), intI, and intII genes among community-acquired (CA) hospital-acquired (HA) MRSA to increase vigilance in the diagnosis and management of suspected infections.

S. aureus isolates recovered from clinical samples were classified into community or hospital-acquired and tested for their antibiotic susceptibility against 19 antibiotics. All isolates were screened for mecA, femA, femB, lukS-PV, lukF-PV, intI, and intII genes. Statistical correlations were carried out.

Out of 338 S. aureus isolates, only 105 were MRSA and classified as 77 CA-MRSA and 28 HA-MRSA. mecA and femA genes were present in all HA-MRSA and CA-MRSA isolates. femB was found in all HA-MRSA and 93.5% of CA-MRSA isolates. PVL genes were detected in 28.6% HA-MRSA isolates and 92.2% CA-MRSA. intI gene was recovered from 60.7% HA-MRSA isolates and 37.7% CA-MRSA isolates while the intII gene recovered from only 10.7% HA-MRSA isolates and 6.5% CA-MRSA.

The high prevalence of MRSA colonizing the groin, axilla, and nose may play a significant role in endogenous infection, re-infection, and also acts as a route for MRSA transmission. mecA and femA genes could be used as a sole and fast step for identification of MRSA, while PVL genes cannot be used as a sole stable marker for CA-MRSA identification.

Hyperglycemia induces cellular senescence in various body cells, such as vascular endothelial cells. Since the vessels are highly distributed in the body and nourish all tissues, vascular damages cause diabetes complications such as kidney failure and visual impairment. Alpha-mangostin is a xanthone found in mangosteen fruit with protective effects in metabolic syndrome and diabetes. This paper has investigated the protective effect of this xanthone against high glucose-induced memory senescence in human vascular endothelial cells (HUVECs) in the presence of metformin, as a positive control. To induce the memory senescence model, HUVECs, after three days incubation with high glucose, were incubated with normal glucose for another three days, and for whole six days, cells were treated with metformin (50 µM) or alpha-mangostin (1.25 µM). On the last day, cell viability by MTT assay, oxidative stress by fluorimetric assay, the number of senescent cells by SA beta-galactosidase staining kit, and secretory interleukin-6 by ELISA kit were measured. SIRT1 and P53 proteins were also evaluated by Western blotting. Metformin and alpha-mangostin significantly increased cell viability, decreased reactive oxygen species, and senescence-associated beta-galactosidase in HUVECs incubated in metabolic memory condition. Generally, metabolic memory increased p53 and acetyl-P53 and decreased SIRT1 proteins in HUVECs, which were reversed by alpha-mangostin and metformin. These data exhibit that alpha-mangostin, comparable to metformin, protects endothelial cells against metabolic memory-induced senescence, which is likely via SIRT1.

Chikungunya virus (ChikV) infection is characterized by persistent infection in joints and lymphoid organs. The ChikV Capsid protein plays an important role in regulating virus replication. In this study, we hypothesized that capsid protein may stimulate dendritic cell (DC) activation and maturation and trigger an inflammatory response in mice. Mice were intraperitoneally injected with capsid protein and examined for changes in immunophenotype in lymph nodes (LNs). Next, DCs were treated with capsid protein or LPS and then expression of maturation markers, cytokine production, and ability to stimulate CD4+ T cells in allo-MLR were analyzed. Injection of mice with capsid protein led to recruitment of myeloid cells and increased activation of T lymphocytes in LNs. Importantly, treatment of DCs with capsid protein prolonged the activation of IKB-α and up-regulated the number of CD11c+CD86+DCs and release of TNF-α and IL-12p70 as well as reducing DC apoptosis, all effects were abolished in the presence of Bay 11-7082. In addition, IL-2 production was higher by CD4+ T cells stimulated with capsid-treated as compared with LPS-induced DCs. The observations revealed that capsid protein participates in the regulation of NF-κB signaling and maturation of DCs.

Bdellovibrio-and-like organisms (BALOs) are predatory prokaryotes that attack and kill other Gram-negative bacteria for growth and reproduction. This study describes the isolation, identification, biological properties, and bacteriolytic activity of the first Bdellovibrio bacteriovorus with a broad prey range from Iran. One BALO strain with high predatory potency was isolated from the rhizosphere soil using Enteropathogenic Escherichia coli as prey. It was identified and designated as Bdellovibrio bacteriovorus strain SOIR-1 through plaque assays, transmission electron microscopy (TEM), Bdellovibrio-specific PCRs, and 16S rRNA gene sequence analysis. Biological characterization and analysis of bacteriolytic activity were also performed. TEM and Bdellovibrio-specific PCRs confirmed that the strain SOIR-1 belongs to the genus Bdellovibrio. Analysis of the 16S rRNA gene sequence revealed its close phylogenetic relationship with strains of Bdellovibrio bacteriovorus. The strain SOIR-1 grew within the temperature range of 25–37 °C and the pH range of 6.0–8.0, with the optimal predatory activity at 30 °C and pH 7.4. It had the highest and lowest bacteriolytic activity toward Shigella dysenteriae and Pseudomonas aeruginosa with a killing rate of 89.66% and 74.83%, respectively.   Considering the hypothesis of bdellovibrios heterogeneity, identification of new isolates contributes to a deeper understanding of their diversity, their ecological roles, and their promising potential as living antibiotics or biocontrol agents. Bdellovibrios with broad bacteriolytic nature has not previously been reported in sufficient detail from Iran. The results of this study showed the great potential of native B. bacteriovorus strain SOIR-1 in the control and treatment of diseases caused by pathogenic Enterobacteriaceae.

This study aimed to investigate the glucose and lipid metabolism improving effect of the total flavonoids from Selaginella tamariscina (Beauv.) Spring (TFST) on db/db mice, and to study its mechanism of action. The db/db mice were divided into 5 groups: the normal group (NC), the diabetic group (DM), the gliclazide group (GZ), the DM+TFST (110 mg/kg), and the DM+TFST (220 mg/kg). The body weight, blood glucose, INS, GC, TC, TG, LDL, and HDL were detected. HE staining was used to observe the liver and pancreas. Urine was tested by UPLC-QTOF-MS to study the metabolic differences of each group, coupled with SIMCA-P13.0 for PCA and OPLS-DA analysis, to identify potential biomarkers, find the metabolic pathway. Western blot was used to examine liver tissue of mice for studying effect of TFST on the PPAR-γ/PI3K/GLU4 pathway. TFST can reduce the weight and levels of TC, TG, and LDL-C, increase the level of GC in blood, and reduce the fat accumulation and inflammation in the liver, and repair the islet cell. 13 biomarkers were identified, they are mainly involved in amino acid metabolism, and purine and pyrimidine metabolism. The results of Western blot show TFST can improve the utilization rate of GLU4 by regulating PPAR-γ and PI3K expression in the liver of db/db mice. TFST can improve glucose and lipid metabolism of DM, which relates to regulation of the PPAR-γ/PI3K/GLU4 signaling pathway, and affect the amino acid metabolism, purine, and pyrimidine metabolism.

We aim to examine possible ammonia lowering effects of mesoporous silica SBA-15 in rats after the common bile duct ligation (BDL). We also evaluate the effect of SBA-15 treatments during 28 days of BDL on locomotion and rearing behavior, as well as on the gene expression of Jnk3 and p38alpha (p38α) mitogen-activated protein kinases in the prefrontal cortex (PFC). SBA-15 was prepared with the hydrothermal method from the surfactant P123 and tetraethyl orthosilicate (TEOS), and calcined at 550 ºC. Then, the product was characterized by FT-IR, XRD, SEM, and BJH-BET techniques. Male Wistar rats in sham control and a group with BDL received saline but another group with BDL received SBA-15 during 28 days of BDL. We examined all groups of rats weekly for locomotion and rearing behavior; then on day 28, we sacrificed all rats, collected the blood sample, and finally dissected the PFC from the whole brain. The SBA-15 treatments had no effect on locomotion but improved rearing behavior on days 7 and 14 of BDL. Biochemical analysis indicated that the SBA-15 treatments in rats with BDL significantly decreased hyperammonemia. The results also revealed that the SBA-15 treatments in rats with BDL significantly restored the decreased Jnk3 gene expression, and increased the p38α gene expression in the PFC. We conclude that SBA-15 can be used as an ammonia lowering agent in hepatic encephalopathy; however, its improving effects on locomotion and neuroinflammation, as well as signaling molecules in the brain need more investigations.

The mechanism via which genistein, the major isoflavone content of soya, adversely influenced placenta and fetal development was evaluated in pregnant laboratory rats. There were control, 2 mg/kg and 4 mg/kg genistein groups of rats with five sub-groups based on gestation termination day. At the end of the experiment, animals were sacrificed by CO2 and cervical dislocation, while plasma and serum were processed and stored. The abdomen was opened and the amniotic fluid was siphoned from the uterine sacs, processed and stored. The embryonic implants were excised, the placenta was separated from the fetus and weighed separately. Placenta homogenate was prepared from the harvested placenta, while the rest were processed for histological studies. Transforming growth factor (TGf-β1) and alkaline phosphatase (ALP) were assayed for in all samples. A significant decrease in the placenta and fetal weights, and a significant decrease in serum and placenta homogenate ALP levels were recorded in genistein groups. There was a reduction in the Trophoblast giant cells population (TGCs). TGCs zone depth, perimeter, and an increase in the placenta and amniotic fluid’s TGf-β1 in all genistein groups at GD-13 towards term, and GD-18 and GD-20, respectively. Maternal plasma TGf-β1 was increased in 2 mg group early in pregnancy while its level significantly decreased in both 2 mg and 4 mg genistein groups at mid-gestation towards GD-19. Genistein aberrant effect on fetal development was via its adverse effect on TGCs proliferation and TGf-β1 activities in the placenta tissue.

Infections by Staphylococcus aureus remain an important health problem. The aims were to detect mecA, staphylococcal cassette chromosome mec (SCCmec), accessory gene regulator (agr), and integrons in S. aureus and to investigate the relationship of agr types with antibiotic resistance of isolates.   In this cross-sectional study, 70 S. aureus isolates were collected between December 2017 and May 2018 from clinical specimens of patients in two hospitals of Sanandaj, western Iran. Susceptibility was determined by disk diffusion for 9 antibiotics and by vancomycin E test. The mecA, classes 1-3 integrons, SCCmec I-V, and agr I-IV were detected by polymerase chain reaction. A P-value<0.05 was considered significant. The most effective antibiotics were linezolid, vancomycin, and trimethoprim-sulfamethoxazole (above 90% sensitivity). Of the 70 isolates, 17.1% were methicillin-resistant S. aureus (MRSA), 8.6% carried class 1 integron, 11.4% carried mecA, 17.1% carried agr I, and 30% carried agr III. SCCmec III and SCCmecV were detected. An association was found between resistance to certain antibiotics and the presence of agr I (P-value<0.05). Conversely, the prevalence of agr III in susceptible strains was higher than non-susceptible strains, and no MRSA isolates belonged to agr III (P-value<0.05). These data suggest that agr activity may influence the resistance of S. aureus to antibiotics. Although the prevalence of mecA and integron was relatively low, the identification of such strains calls for serious health concerns; thus highlights the need to monitor drug resistance in S. aureus.

Here we investigated the regulation of Th17 and Treg cells in orthodontic tooth movement during periodontal inflammation. Fifty-six SD rats were divided into a control (24 rats) and a tooth movement group during the recovery stage of periodontitis (RM group, 32 rats). Periodontitis was established by silk ligation and local injection of LPS. Orthodontic tooth movement was achieved by nickel-titanium springs on the maxillary first molars. The proportions of Th17 cells and Treg cells were evaluated by flow cytometry. Gene expression of ROR-γt and Foxp3 was determined by real-time PCR. Expression of ROR-γt, Foxp3, RANK, RANKL, and OPG was detected by immunohistochemical staining. Osteoclasts were detected by TRAP staining. Relationships between Th17/Treg cells, osteoclasts, and related factors were estimated by correlation and regression analysis. During orthodontic tooth movement in the recovery stage of periodontitis, the proportion of Th17 cells, ROR-γt, RANK, osteoclasts, and the RANKL/OPG ratio increased and then decreased. The proportion of Treg cells and Foxp3 increased, then decreased, and increased again. Levels of RANKL and OPG increased, then decreased, then increased, and finally decreased. The Th17/Treg ratio initially decreased, then increased, and decreased again. Th17 cells were positively correlated with RANK and RANKL, the RANKL/OPG ratio, and counts of osteoclasts. Treg cells were negatively correlated with RANK expression and numbers of osteoclasts. The Th17/Treg ratio was positively correlated with RANK expression and numbers of osteoclasts. Under periodontal inflammation conditions, the Th17/Treg ratio might regulate orthodontic tooth movement through changing osteoclasts metabolism.

Human gastrointestinal tract harbors a variety of bacteria with vital roles in human health. Bacteroides fragilis is considered one of the dominant constituents of gut microflora which can act as an opportunistic pathogen leading to various diseases, including colon cancer, diarrhea, uterine and intrathecal abscesses, septicemia, and pelvic inflammation. In this study, multiple locus variable number of tandem repeats analysis (MLVA) was performed to genetically differentiate 50 B. fragilis isolates. Eight suitable tandem repeats (TRs) were selected by bioinformatics tools and were then subjected to PCR amplification using specific primers. Finally, MLVA profiles were clustered using BioNumerics 7.6 software package. All VNTR loci were detected in all isolates using the PCR method. Overall, B. fragilis isolates were differentiated into 27 distinct MLVA types. The highest diversity index was allocated to TR1, TR2, TR5, TR6, and TR8; with this taken into account, strain type 14 was the most prevalent with 12 strains belonging to this type. Clustering revealed three major clusters of A, B, and C. With regards to the pathogenicity of B. fragilis and the outcomes of infections related to this microorganism, it is imperative to study this microorganism isolated from both patients and healthy individuals. This study aimed at evaluating the efficiency of MLVA for the genetic differentiation of B. fragilis. The results of this study indicate the promising efficiency of MLVA typing for cluster detection of this bacterium.

Prevention of inflammation in early stages will be useful in maintaining vitality of the organism. The objective of this study was to evaluate the effects of doxycycline (DOX) or meloxicam (MLX) monotherapy and combination therapy on the levels of inflammatory mediators in the brain tissues of rats with Escherichia coli lipopolysaccharide (LPS)-induced brain inflammation. Seventy-eight rats were divided into the following groups: control (n=6), LPS (0.5 µg/10 µl intracranial) (n=18), LPS (0.5 µg/10 µl intracranial)+DOX (40 mg/kg intraperitoneal) (n=18), LPS (0.5 µg/10 µl intracranial)+MLX (2 mg/kg intraperitoneal) (n=18) and LPS (0.5 µg/10 µl intracranial)+DOX (40 mg/kg intraperitoneal)+MLX (2 mg/kg intraperitoneal) (n=18) groups. Brain tissues were harvested from all rats in the control group and from six rats each in the four experimental groups at 1, 3 and 6 hr under anaesthesia. The levels of tumor necrosis factor α (TNFα), interleukin 4 (IL-4), IL-6, IL-10, IL-17, brain-derived neurotrophic factor (BDNF), matrix metalloproteinase 3 (MMP-3), tissue inhibitor of metalloproteinase 3 (TIMP-3) and cyclooxygenase 2 (COX-2) in the brain tissues were measured using ELISA kits with ELISA device. LPS administration increased proinflammatory cytokines (TNF, IL-6, IL-17), and MMP-3 levels and decreased anti-inflammatory cytokines (IL-10, IL-4), and BDNF levels. The lowest TNFα levels were detected in the LPS+MLX group (P<0.05). All the drug treatment groups showed decreased IL-17 and COX-2 levels compared to the LPS groups. DOX or MLX monotherapy exerts neuroprotective effects against brain inflammation by decreasing proinflammatory cytokine levels and by increasing anti-inflammatory cytokines levels.

Emergence of resistant tumor cells to the current therapeutics is the main hindrance in cancer treatment. Combination therapy, which mixes two or more drugs, is a way to overcome resistant problems of cancer cells to current treatments.  Nanobodies are promising tools in cancer therapy due to their high affinity as well as high penetration to tumor sites. Here, the inhibitory effect of mixtures of two nanobodies (anti-vascular endothelial growth factor (VEGF) and anti-neuropilin-1 (NRP-1) nanobodies) on tube formation of human endothelial cells in vitro and ex vivo were analyzed. Results showed that combination of two drugs significantly inhibited proliferation and tube formation of human endothelial cells. In addition, mixtures of two nanobodies inhibited angiogenesis in chick chorioallantoic membrane (CAM) assay efficiently compared with each individual nanobody. Results highlight the efficacy of combination therapy of cancer compared with mono-therapy and promises development of novel anti-cancer therapeutics based on nanobodies targeting two or more targets of tumor cells.

To investigate the effect of Ginkgo biloba extract EGb761 in early brain injury (EBI) after subarachnoid hemorrhage (SAH) and its mechanism. The SAH rat model was constructed and pre-treated with EGb761.The neurological function, severity of SAH, water content of brain tissue, damage degree of the blood-brain barrier, related indexes of oxidative stress, and the level of inflammatory cytokines were compared among the groups. The expression of TXNIP/NLRP3 signaling pathway-related proteins in brain tissues was detected by Western blot. After SAH modeling, the neurological function score was significantly reduced, the degree of brain injury, levels of oxidative stress, inflammatory factors, expression of NLRP3 and TXNIP were all increased. Compared with the SAH rats, the neurological function score of rats pre-treated by EGb761 was higher, the degree of brain injury, levels of oxidative stress and inflammatory factors, expression of NLRP3 and TXNIP were all lower. EGb761 could protect neurological injury after SAH and its mechanism may be that EGb761 could inhibit the activation of the TXNIP/NLRP3 signaling pathway and inflammatory reaction after oxidative stress.

Glutamate is the most widespread neurotransmitter in the central nervous system and has several functions as a neuromodulator in the brain although in pathological conditions like ischemia it is excessively released causing cell death. Under physiological conditions, glutamate is rapidly scavenged from the synaptic cleft by excitatory amino-acid transporters (EAATs). An imbalance in glutamatergic neurotransmission could influence the expression of glutamate transporters and is a pathological feature in several neurological disorders. It has been shown that estrogen and progesterone act as neuroprotective agents after brain injury. This study aims to investigate the role of hormone therapy after middle cerebral artery occlusion (tMCAO) in the expression of GLT-1 and EAAT3 as glutamate transporters. Middle cerebral artery occlusion technique was performed in Wistar rats in order to induce focal cerebral ischemia. Estrogen, progesterone, and a combination of both hormones were injected subcutaneously in the early minutes of reperfusion. Sensorimotor functional tests were performed and infarct volume was calculated by TTC staining of brain section. Gene and protein expression of EAAT3 and GLT-1 were evaluated by RT-PCR, immunoblotting, and immunohistochemistry. Behavioral scores were increased and infarct volume was reduced by hormone therapy. RT-PCR, immunoblotting, and immunohistochemistry data showed that the expression of GLT-1 and EAAT3 increased after ischemia. Also, estrogen and progesterone treatment enhanced mRNA and protein expression levels of GLT-1 and EAAT3 compared with ischemia. Steroids may protect brain tissue against ischemia-induced tissue degeneration by decreasing extracellular glutamate levels through the induction of glutamate transporters.

The aims of the present study were to investigate the expression of calcium sensing receptor (CaSR) at different times in acute myocardial infarction (AMI) rat myocardial tissue after mouse embryonic stem cells (mESCs) transplantation treatment and to assess its effects on apoptosis and oxidative stress of cardiomyocytes. The AMI rats were treated with mESCs, Calindol (a CaSR agonist) and Calhex231 (a CaSR inhibitor). Serum measurements, Echocardiographic analysis and TUNEL assay were performed. Myocardial ultrastructure changes were viewed by electron microscopy. Additionally, western blotting was used to detect the protein expressions. Compared to the sham group, it was found that the expression levels of CaSR, caspase-3, cytoplasmic cytochrome C (cyt-C) and Bcl2-associated x (Bax), and the levels of Malondialdehyde (MDA) were significantly increased in both AMI and AMI + mESCs + Calindol groups with the development of myocardial infarction. Furthermore, the ultra-microstructure of cardiomyocyte was highly damaged, the expression levels of mitochondrial cyt-C and B-cell lymphoma 2 (Bcl-2) were significantly decreased, and there was decreased activity of superoxide dismutase (SOD). However, the combination of Calhex231 and mESCs transplantation could inhibit these changes. Our results suggested that CaSR expression in myocardial tissue of AMI rats was increased over time, and that Calhex231 could enhance the efficacy of ESCs transplantation for the treatment of AMI, which would be a new therapeutic strategy for the treatment of AMI.

Thymoquinone (TQ) has valuable medical properties like anticancer effects. Development of multidrug resistance (MDR) phenotype is one of the most important factors in failure of cancer chemotherapy. The aim of this study was to evaluate the mode of interaction of TQ and MDR1, a major MDR-related protein in gastric cancer drug resistant EPG85-257RDB cells, and its parental non-resistant EPG85-257 cells. MTT assay was used to assess the effects of TQ and doxorubicin (DOX) on cell viability of tested cell lines and TQ effect on pump performance. HPLC analyses were used to measure the input and output of TQ in EPG85-257RDB cells. Molecular docking studies were used to identify interactions between TQ and MDR1. TQ inhibited cell viability in a time and concentration-dependent manner. Co-treatment of the cells with TQ and DOX did not significantly affect the amount of cell viability in comparison with DOX treatment alone. The HPLC analyses showed that more than 90% of TQ entered to EPG85-257RDB during 1 hr of treatment with TQ, but it was unable to exit from the cells. Moreover, there was no difference between influx and efflux amount of TQ in cells with inhibited and non-inhibited MDR1 transporters. Molecular docking studies revealed that TQ had a higher inhibitory constant to bind to active site of MDR1 protein as compared to specific inhibitor (verapamil) and substrate (vinblastine) of this transporter. These results proposed that TQ does not work as an inhibitor or a substrate of MDR1 transporter.