Application and development of a TaqMan-based real-time PCR assay for rapid detection of snakehead vesiculovirus.

Snakehead vesiculovirus (SHVV) is one of the primary pathogens responsible for viral diseases in the snakehead fish. A TaqMan-based real-time PCR assay was established for the rapid detection and quantification of SHVV in this study. Specific primers and fluorescent probes were designed for phosphoprotein (P) gene, and after optimizing the reaction conditions, the results indicated that the detection limit of this method could reach 37.1 copies, representing a 100-fold increase in detection sensitivity compared to RT-PCR. The specificity testing results revealed that this method exhibited no cross-reactivity with ISKNV, LMBV, RSIV, RGNNV, GCRV, and CyHV-2. Repetition experiments demonstrated that both intra-batch and inter-batch coefficients of variation were not higher than 1.66%. Through in vitro infection experiments monitoring the quantitative changes of SHVV in different tissues, the results indicated that the liver and spleen exhibited the highest viral load at 3 poi. The TaqMan-based real-time PCR method established in this study exhibits high sensitivity, excellent specificity, and strong reproducibility. It can be employed for rapid detection and viral load monitoring of SHVV, thus providing a robust tool for the clinical diagnosis and pathogen research of SHVV.

Identification and characterization of a highly virulent Citrobacter freundii isolate and its activation on immune responses in largemouth bass (Micropterus salmoides).

Citrobacter freundii, a common pathogen of freshwater fish, causes significant commercial losses to the global fish farming industry. In the present study, a highly pathogenic C. freundii strain was isolated and identified from largemouth bass (Micropterus salmoides). The pathogenicity and antibiotic sensitivity of the C. freundii strain were evaluated, and the histopathology and host immune response of largemouth bass infected with C. freundii were investigated. The results showed that C. freundii was the pathogen causing disease outbreaks in largemouth bass, and the infected fish showed typical signs of acute hemorrhages and visceral enlargement. Antimicrobial susceptibility testing showed that the C. freundii strain was resistant to Kanamycin, Medimycin, Clindamycin, Penicillin, Oxacillin, Ampicillin, Cephalexin, Cefazolin, Cefradine and Vancomycin. Histopathological analysis showed different pathological changes in major tissues of diseased fish. In addition, humoral immune factors such as superoxide dismutase (SOD), catalase (CAT) and lysozyme (LZM) were used as serum indicators to evaluate the immune response of largemouth bass after infection. Quantitative real-time PCR (qRT-PCR) was performed to investigate the expression pattern of immune-related genes (CXCR1, IL-8, IRF7, IgM, CD40, IFN-γ, IL-1ß, Hep1, and Hep2) in liver, spleen, and head kidney tissues, which demonstrated a strong immune response induced by C. freundii infection in largemouth bass. The present study provides insights into the pathogenic mechanism of C. freundii and immune response in largemouth bass, promoting the prevention and treatment of diseases caused by C. freundii infection.

Lusianthridin Exerts Streptozotocin-Induced Gestational Diabetes Mellitus in Female Rats via Alteration of TLR4/MyD88/NF-κB Signaling Pathway.

Gestational diabetes mellitus (GDM) is characterized via enhanced the glucose intolerance in the pregnant women, which further lead the expansion of gestational hypertension, hepatic damage, pre eclampsia and renal damage. Lusianthridin is the active phytoconstituent of Dendrabium venustu and exhibited the antioxidant and anti-inflammatory effects. In this protocol, we examined the GDM protective effect of lusianthridin (LSD) against streptozotocin (STZ) induced GDM in the female rats. Single intraperitoneal injection of STZ (40 mg/kg) was used for the induction of diabetes in the pregnant female rats. The rats were orally treated with the LSD (10, 20 and 40 mg/kg, body weight) for 18 days and blood glucose level, body weight and plasma insulin were estimated at regular time intervals. at end of the study, fetal weight, placental weight, number of live and dead fetuses were estimated. The antioxidant, lipid and cytokines level were also estimated. GDM rats treated with LSD remarkably improved the body weight of female rats along with fetal weight and suppressed the placental weight. LSD enhanced the live fetuses and suppressed the dead fetuses with reduction of reduced the dead ratio. LSD considerably suppressed the glucose level and improved the insulin level and suppressed the HOMA-IR. LSD significantly (p < 0.001) increased the level of hemoglobin, glycogen and suppressed the level of glycalated hemoglobin. LSD significantly (p < 0.001) altered the level of lipid parameters and inflammatory cytokines. LSD altered the level of antioxidant parameters in the liver and pancreas tissue. LSD significantly (p < 0.001) decreased the mRNA expression of troll like receptor (TLR)4, myeloid differentiation primary response 88 (MyD88), Nuclear factor kappa B (NF-κB)p65 and nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing-3 (NLRP3), respectively. The results suggest that LSD has a protective effect on GDM in female rats induced by STZ, possibly through reducing the activity of the TLR4/MyD88/NF-κB signaling pathway.

Novel synthesized zinc oxide nanoparticles loaded alginate-chitosan biofilm to enhanced wound site activity and anti-septic abilities for the management of complicated abdominal wound dehiscence.

Wound dehiscence is a surgical complication and its management is inevitable because 25% to 35% of patients suffered from post laparotomy wound dehiscence. The excellent biodegradability and biocompatibility of chitosan and alginate have provided ample space for future developments in biomedical applications. Hence, the present work is directed towards the synthesis of robust biofilm made up of chitosan (CS), zinc oxide (ZnO) nanoparticles and Alginate (Alg). Chitosan and alginate were used for their pore forming ability, and ZnO is for its antibacterial action. The proposed biofilm was characterized with different characterization techniques such as Fourier Transform Infrared (FTIR) spectroscopy, UV-vis spectroscopy, X-ray Diffraction (XRD), Scanning Electron Microscopy (SEM) and Transmission Electron microscopy (TEM) analyses. FTIR results inferred the strong interaction between the three components. The surface morphology of ZnO-CS/Alg. biofilm was exhibited as the spherical shaped nanoparticles which are firmly anchored on the polymer matrix. TEM analysis also confirmed the formation of biofilm. The XRD analysis confirmed the presence of ZnO in the biopolymer. The line broadening suggests that the crystallize size is in few nanometers. The average crystallite size was estimated as 50 nm using Scherrer formula. The antibacterial activity of the biofilm was successfully established against bacterial pathogens. Therefore, the developed materials have a potential play as antimicrobial role for the abdominal wound healing and biomedical fields.

Development of biosynthesized silver nanoparticles based formulation for treating wounds during nursing care in hospitals.

Silver nanoparticles (AgNPs) have been emerged as significant wound healing agents because of their improved mechanical properties. However, the green synthesized silver nanoparticleshave reported significant wound healing action in Albino rats which was validated by the measurement of wound closure rate. Silver nanoparticles were efficiently synthesized using Euphorbia milii leaf extract. The UV-visible spectra recorded the effect of the reaction time on AgNPs synthesis and was indentified that the peak became shaper with an increase in time, which corresponds to increase in the number of nanoparticles formed from the reduction of silver ions present in the aqueous solution. X-ray diffraction technique and corresponding XRD patterns confirmed the biphasic nature of the biosynthesized silver nanoparticles. However, low magnification TEM images presented monodispersed AgNPs with their size ranging from 20 to 30 nm while SAED diffraction pattern disclosed their crystalline nature. Furthermore, the wound healing activity of AgNPs was examined through the excision wound model by measuring the rate of wound closure and Group II (treated with 10% Ointment base with biosynthesized AgNPs) revealed significant wound healing activity over Control group and Group I (treated with Standard Nitrofurazone ointment) in Albino rats.

MiR-661 contributed to cell proliferation of human ovarian cancer cells by repressing INPP5J expression.

Accumulating evidence has emerged important roles for microRNAs (miRNAs) participating in oncogenesis and growth of various cancers. We hypothesized that miR-661 played an important role in cell growth of ovarian cancer. Here, we found miR-661 was upregulated in human ovarian cancer cell lines and clinical tumor tissues. Our results revealed that miR-661 directly targeted INPP5J, thereby acting as tumor promoter in ovarian cancer cells by blocking cell proliferation. Importantly, we identified miR-661 as a positive regulator of INPP5J-induced AKT pathway. Taken together, our study sheds light onto the role of miR-661 as tumor promoter by targeting the INPP5J gene, and then promoting cell proliferation of ovarian cancer.