Moderate Toxicity of Potential Boron-containing Therapeutic, Dipotassium-trioxohydroxytetrafl uorotriborate - K2 (B3 O3 F4 OH) in Rats and Mice

Abstract Biological activity of boron-containing compounds (BCCs) has been well-known. Growing interest and numerous applications for BCCs have been reported. Boron and boron-containing acids show low acute toxicity in mammals but data on halogenated boroxine (HB) - dipotassium-trioxohydroxytetrafluorotriborate, K2(B3O3F4OH) acute toxicity have not been reported before. This compound, characterized as a potential therapeutic for skin changes, exhibits no observable genotoxicity in doses lower that 0.1 mg/ml in vitro and 55 mg/kg in vivo. It has also been confirmed as an antitumour agent both in vitro and in vivo as well as an inhibitor of enzymes involved in antioxidant mechanisms. The aim of this study was to assess the acute toxicity of HB and to determine the maximum tolerated dose as well as a dose free of any signs of toxicity in different test organisms. Acute toxicity of HB was tested in Sprague-Dawley and Wistar rats and BALB/c mice after single parenteral application of different doses. We determined doses free of any sign of toxicity and LD50 after single dose administration. LD50 of HB ranges from 63 to 75 mg/kg in different test models, meaning that HB shows moderate toxicity

A modified model of glaucoma filtering surgery in Sprague-Dawley rats

Animal models are useful in glaucoma research to study tissue response to wound healing. Smaller animals such as rats offer additional advantages in terms of availability of detection antibodies and microarrays with cheaper maintenance costs. In this study, we describe a glaucoma filtering surgery (GFS) model in adult Sprague–Dawley rats by performing a sclerostomy using a 26?G needle and additionally placing a silicone tube (27 G) connecting the anterior chamber to the subconjunctival space to maintain a patent fistula for the flow of aqueous humor, thus providing a more definitive bleb. This technique will be useful in identifying and modifying newer targets in the wound healing process in order to improve surgical outcomes following GFS

Effects of 275 nm and 310 nm ultraviolet irradiation on bone metabolism in ovariectomized osteoporotic rats / 北京大学学报(医学版)

OBJECTIVE@#To investigate the effect of 275 nm and 310 nm ultraviolet irradiation on ovariectomized rats' bone metabolism.@*METHODS@#Twenty four 3-month-old female Sprague-Dawley (SD) rat were randomly divided into control group, sham operated group, 275 nm ultraviolet (UV) irradiation group and 310 nm UV irradiation group. Each group contained 6 rats. The rats in the two irradiation groups were treated with bilateral ovariectomy. The rats in sham operated group received sham operation (They were given the same back incision and a bit of par-ovarian fat were removed). Control group received no disposition. About 24 weeks after operation, all the rats received detailed bone mineral density (BMD) detection again. Detection regions include cervical vertebra, lumbar vertebra, proximal femur, mid femur and distal femur. Next, osteopenia rats in 275 nm irradiation group were UV irradiated 275 nm with fixed illumination intensity (15 μW/cm2) everyday for 16 weeks. The osteopenia rats in 310 nm irradiation group were UV irradiated 310 nm with fixed illumination intensity (15 μW/cm2) everyday for 16 weeks. The backs of the rats were shaved regularly as irradiation area (6 cm×8 cm). After 16-week irradiation, all the rats' BMD of cervical vertebra, lumbar vertebra, proximal femur, mid femur and distal femur were measured. At the end of the trial, all the rats' blood specimens were obtained and serum 25(OH)D, procollagen type Ⅰ N-peptide (PINP) and osteocalcin (OC) were measured.@*RESULTS@#Compared with control group [(238.78±26.74) mg/cm3], the BMD of the whole body were significantly lower in 275 nm [(193.34±13.28) mg/cm3] and 310 nm [(191.19±18.48) mg/cm3] irradiation groups (P=0.002, P=0.001). There were no significant difference between sham operated group [(227.20±14.32) mg/cm3] and control group. After 16-week ultraviolet irradiation, the BMD of the whole body were significantly increased in 275 nm [(193.34±13.28) mg/cm3 vs. (221.68±25.52) mg/cm3, P=0.005] and 310 nm groups [(191.19±18.48) mg/cm3 vs. (267.48±20.54) mg/cm3, P < 0.001] after corresponding irradiation. The BMD of the four body regions (lumbar vertebra, proximal femur, mid femur and distal femur) had significantly increased after irradiation in 275 nm irradiation group. For 310 nm irradiation group, the BMD in cervical vertebra, lumbar vertebra, proximal femur, mid femur and distal femur also had increased significantly after 310 nm ultraviolet irradiation. The concentration of serum 25(OH)D and OC was higher in 275 nm irradiation group than in control group [(46.78±5.59) μg/L vs. (21.32±6.65) μg/L, P=0.002;(2.05±0.53) U/L vs. (1.32±0.07) U/L, P=0.022]. Compared with the control, the concentration of serum 25(OH)D [(58.05±12.74) μg/L], OC [(2.04±0.53) U/L] and PINP [(176.16±24.18) U/L] was significantly higher (P < 0.001, P=0.015, P=0.005) in 310 nm irradiation group. However, there were no significantly difference between sham operated group and the control.@*CONCLUSION@#Both 275 nm and 310 nm ultraviolet could improve rats' vitamin D synthesis. Both 275 nm and 310 nm ultraviolet could improve osteopenia rats' bone condition. The irradiation of 310 nm might be more effective on bone condition improvement.

<i>In vitro</i> isolation and culture of SD rat retinal ganglion cells and establishment of a high-glucose model / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To isolate the retinal ganglion cells(RGCs)of neonatal Sprague-Dawley(SD)rats <i>in vitro</i>, and to establish the RGCs primary culture method and high glucose model of neonatal SD rats.<p>METHODS: The retinal tissues of SD rats from 1-3d as the materials were taken, from which the RGCs were isolated and purified for primary culture. Toluidine blue and immunofluorescence staining methods were adopted to identify the cultured cells. After 48-72h of continuous culture, RGCs were randomly divided into 6 groups and cultured in different glucose concentrations of 5.5mmol/L(normal control group), 20mmol/L, 25mmol/L, 30mmol/L, 35mmol/L and 40mmol/L for 24, 48 and 72h, respectively. Finally, the CCK8 method and TUNEL method were adopted to determine the cell survival rate and apoptosis rate. <p>RESULTS: The primary RGCs purified and cultured <i>in vitro</i> represented typical cell morphology and grew well, the cells were confluent and aggregated in small lamellar manner, while the axons crisscrossed into a network, in addition, cell halo could be seen around the cell body. Nissl bodies with clear structure were found in the cytoplasm of toluidine blue stained cells, the percentage of neurons was more than 95%. RGCs specific antibodies Thy-1 and Brn-3a were employed to identify the purified cells <i>in vitro</i>, and the positive rate reached more than 90%. The CCK8 results showed that the survival rate of cells decreased(OD value decreased)with the increase of culture time and glucose concentration. When the cells were treated with different glucose concentrations for 24h, the OD values of each group were lower than those of the normal control group, but there was no significant difference between the OD values of each group and the normal control group(all <i>P</i>>0.05). With the extension of culture time, the OD values of 35mmol/L and 40mmol/L glucose concentration intervention RGCs 48h, 30mmol/L, 35mmol/L, 40mmol/L intervention RGCs 72h were significantly lower than those of the normal control group, the difference was statistically significant compared with the normal control group(all <i>P</i><0.05). TUNEL results revealed that the apoptosis rate of RGCs increased with the increase of glucose concentration and time, among them, the apoptosis rate of RGCs cultured in glucose concentration of 30mmol/L, 35mmol/L and 40mmol/L for 48h and 72h was significantly statistical different from that of normal control group(all <i>P</i><0.05). <p>CONCLUSION: The RGCs primary culture method established in this study is capable of separating typical RGCs with high purity. With the increase of glucose concentration in the medium, the survival rate of RGCs have been decreased while the apoptosis rate increased. Notably, the 35mmol/L glucose intervention for 48h can be employed as the optimal intervention concentration and time to effectively induce RGCs to establish the high glucose model.

Acute toxicity evaluation of homeopathic preparation of Gymnema sylvestre and analysis of its chemical constituents

In many countries, homeopathic preparation is believed to be an effective medicine for various ailments.However, there are limited scientific evidences in regard to its usage, safety, and efficacy. It is necessary toupdate this age-old scientific wisdom in different aspects, including pharmacologic and therapeutic potentials.In this study, we assessed the safety profile of homeopathic preparation of Gymnema sylvestre (HPGS). Itschemical constituents were deciphered using LCMS approaches. HPGS was subjected to an acute toxicitystudy (OECD-423 guidelines) using Sprague Dawley rats. The administration of HPGS did not produce anytoxic symptoms or show mortality at the dose level of 300 mg/kg body weight. Phytochemical analysis revealedthat HPGS contained alkaloids, saponins, and flavonoids. These results demonstrated the non-toxic nature ofHPGS in vivo, suggesting a long-term usage in clinical practices when administered orally

Ensayo de inmunogenicidad y toxicidad aguda del candidato vacunal contra el cólera vax-COLER® / Immunogenicity and acute toxicity assay of the vaccine candidate against cholera vax-COLER®

Para el control y prevención del cólera humano se han llevado a cabo diferentes estrategias, entre las cuales, la vacunación es una de las medidas más eficaces. La evaluación preclínica de candidatos vacunales requiere de la demostración de la seguridad de los mismos, para lo cual los estudios toxicológicos son determinantes, al ser obligatorios y altamente regulados. Este estudio tuvo como objetivo demostrar la relevancia de las ratas Sprague Dawley como biomodelo a través de su respuesta inmunológica al candidato vacunal contra el cólera, vax-COLER®, utilizando la técnica de determinación de anticuerpos vibriocidas. Además, evaluar los efectos toxicológicos locales y sistémicos por la administración de una dosis de vax-COLER® a través de la evaluación de síntomas, del consumo de agua y alimentos, el peso corporal y estudios anatomopatológicos. La vacuna vax-COLER® resultó inmunogénica y no evidenció síntomas ni muertes, no hubo cambios en el peso corporal y los consumos de agua y alimentos se comportaron de forma similar entre todos los grupos. Los estudios anatomopatológicos solo mostraron cambios a nivel histológico en los ganglios linfáticos mesentéricos y placas de Peyer de los animales vacunados, con presencia de hiperplasia de los folículos secundarios subcapsulares, hallazgo que difirió significativamente con el resto de los grupos. Se concluye que la vacuna vax-COLER® es inmunogénica en ratas Sprague Dawley, demostrando la relevancia del biomodelo para la evaluación de la seguridad preclínica y que la aplicación de una dosis no produjo efectos tóxicos agudos generales ni locales(AU)

Different strategies have been carried out for the control and prevention of human cholera. Vaccination is one of the most effective strategies. Preclinical evaluation of vaccines needs to prove their safety; whereby toxicological studies are decisive. They are mandatory and highly regulated. This study was aimed to demonstrate the relevance of Sprague Dawley rats as a biomodel, through the immunological response to vax-COLER® cholera vaccine, using the technique of determination of vibriocidal antibodies. In addition, local and systemic toxicological effects were evaluated after administration of a dose of vax-COLER®; through the evaluation of symptoms, water and food consumption, body weight and anatomopathological studies. The vax-COLER® vaccine was immunogenic and showed no symptoms or deaths. No changes in body weight were detected, and food and water consumption were similar among all groups. The anatomopathological studies showed histological changes in the mesenteric lymph nodes and Peyer's patches of the vaccinated animals, with hyperplasia of the subcapsular secondary follicles, finding that differed significantly from the rest of the groups. It is concluded that vax-COLER® vaccine is immunogenic in Sprague-Dawley rats, demonstrating the relevance of the biomodel for the evaluation of preclinical safety, as well as that the application of a single dose did not produce acute general or local toxic effects(AU)

Evaluation Of Antihyperlipidemic Activity Of Ethanolic Extract Of Crossandra Infundibuliformis Leaves And Stems On High Fat Diet Induced Hyperlipidemia

Hypercholesterolaemia is the state been depicted by an expanded greasy substances called lipids, rise in plasma TCs and TGs levels it is additionally called hyper lipoproteinemia. The present study was designed to investigate the hypolipidemic effect of ethanolic extract of Crossandra infundibuliformis leaves and stem (ECILS) in high fat diet (cholesterol 2%, sodium cholate 1%, sucrose 48%, peanut oil, methionine 4%, and 47% normal laboratory feed) . The rats feed with High-fat diet containing (cholesterol 2%, sodium cholate 1%, sucrose 48%, peanut oil, methionine 4%, and 47% normal standard laboratory feed) for 1 month, and then they are checked for the blood parameter levels like TC, TGs, LDL, VLDL, and HDL. Ethanolic extracts at low dose (100 mg/kg), significantly reduced the levels of TC, TGL, LDL, VLDL & increased the levels of HDL and reduced the body weights on 30st day, and at medium, high doses it reduced the levels of TC, TGL, LDL, VLDL & increased the levels of HDL and reduced the body weights.

Efecto del ultrasonido de baja intensidad (USBI) sobre el movimiento dentario ortodóncico: estudio in vivo en ratas Sprague-Dawley / Effect of low intensity ultrasound (USBI) in the orthodontic dental movement: in vivo study in Sprague-Dawley rats

Este estudio se realizó con el propósito de analizar el efecto de la aplicación de ondas de USBI sobre la velocidad y distancia de movimiento dentario, y su efecto a nivel histológico en ratas Sprague-Dawley. Veintitrés ratas macho Sprague-Dawley fueron parte del experimento, las que fueron divididas aleatoriamente en tres grupos. Cinco ratas fueron utilizadas como control sin movimiento dentario ortodóncico (MDO), nueve ratas fueron sometidas a una aplicación de fuerza ortodóncica sin aplicación de USBI y nueve fueron sometidas a una aplicación de fuerza ortodóncica con USBI. Estas fuerzas ortodóncicas fueron realizadas entre el aspecto mesio vestibular del primer molar superior izquierdo y el incisivo central superior, traccionando el molar hacia mesial, aplicando una fuerza aproximada de 50 gramos durante 21 días, debido a problemas con la sedación estos dos últimos grupos quedaron compuestos de seis y ocho ratas respectivamente. En el grupo MDO sin USBI, se registró una reducción del espacio entre molares e incisivos de 2 mm. En el grupo MDO con USBI dicha reducción fue de 3.4 mm, siendo esta diferencia estadísticamente significativa. Histológicamente el grupo MDO c/USBI mostró un perímetro vascular y espacios medulares aumentados respecto de los otros grupos, no obstante, no se encontraron diferencias significativas en la cantidad de osteoclastos. Fue posible observar y demostrar cuantitativamente que el USBI es una forma de estimulación mecánica eficiente para la aceleración del MDO sobre un período experimental de 21 días, donde se observaron diferencias significativas entre el grupo control, el grupo MDO sin USBI y el grupo MDO con USBI en la magnitud del movimiento dentario. Se observó una mayor vascularidad en el grupo con USBI, expresando perímetros vasculares de mayor tamaño, esto pudo deberse al efecto estimulatorio del ultrasonido sobre la angiogénesis.

The objective of this study was to analyze the effect of USBI wave application on the speed and distance of dental movement, and its effect at histological level in SpragueDawley rats. In this study twenty-three male Sprague-Dawley rats were randomly divided into three groups. Five rats were used as control without orthodontic tooth movement (MDO), nine rats were subjected to an orthodontic force application without USBI application, and nine were subjected to an orthodontic force application with USBI. The orthodontic forces were performed between the vestibular mesio aspect of the first upper left molar and upper central incisor, pulling the molar mesially, applying an approximate force of 50 grams for 21 days. In light of sedation problems these last two groups were composed of six and eight rats respectively. In the MDO group without USBI, there was a reduction in the space between molars and incisors of 2 mm. In the MDO group with USBI this reduction was 3.4mm, this difference being statistically significant. Histologically, the MDO c / USBI group showed a vascular perimeter and enlarged spinal spaces with respect to the other groups, however, no significant differences were found in the amount of osteoclasts. It was possible to quantitatively demonstrate that USBI is a form of efficient mechanical stimulation for MDO acceleration over an experimental 21-day period, where significant differences were observed in tooth movement between the control group, the group without USBI and the group with USBI. A greater vascularity was observed in the group with USBI, expressing larger vascular perimeters, which could be due to the stimulatory effect of ultrasound on angiogenesis.

Effects of 1,25(Oh)2d3 on microrna-130b and transforming growth factor beta 1 in renal tissue of rat models of diabetic nephropathy / 中国组织工程研究

BACKGROUND: 1,25(OH)2D3 plays an important regulatory role in the development of diabetic nephropathy. OBJECTIVE: To explore the effect of 1,25(OH)2D3 on the expression of microRNA-130b and transforming growth factor pi in kidney of diabetic nephropathy rats. METHODS: The study was approved by the Laboratory Animal Ethical Committee of Laboratory Animal Center of Xinjiang Medical University. Twenty-five clean Sprague-Dawley rats were randomly divided into normal control group, diabetic nephropathy+1,25(OH)2D3 group and diabetic nephropathy+peanut oil group. The latter two groups were given calcitriol (1,25(OH)2D3, active vitamin D3, 0.03 jg/kg

Improvement for the construction of renal interstitial fibrosis model with unilateral ureteral obstruction in rats / 中国组织工程研究

BACKGROUND: Unilateral ureteral obstruction (UUO) is a traditional surgical method that can cause renal interstitial fibrosis in rats in a short period of time (1-2 weeks), but it can develop many postoperative complications and result in a high mortality. OBJECTIVE: To optimize and improve the UUO operation in rats for reducing the incidence of postoperative complications and improving the survival rate of animals, and to detect the pathophysiological indicators of rats modeled by UUO, providing background data for the basic research on functional pharmacology. METHODS: Twenty male Sprague-Dawley rats were randomly divided into four groups, followed by traditional UUO ligation, improved UUO ligation, undissociated ureteral ligation, and opening the abdominal cavity with no ureter ligation (sham group), respectively. At 14 days postoperatively, the successful rate of renal interstitial fibrosis model, mortality rate, and the incidence of postoperative complications were compared between groups. Another 28 Sprague-Dawley rats, half male and half female, were randomly divided into improved UUO group and control group. Physiological indexes, including urine analysis, blood cells analysis, blood biochemical analysis, were tested as background information. Besides, the pathological changes of kidney tissues were compared between groups using hematoxylin-eosin staining and Masson staining at 14 and 21 days postoperatively. RESULTS AND CONCLUSION: Compared with the undissociated ureteral ligation group, the successful rate of modeling was higher in the traditional and improved UUO groups. The improved UUO surgery with double ligation of the middle ureter and no cutting of the middle ureter was characterized by easier operation, smaller surgical wound, faster modeling and higher successful rate of renal interstitial fibrosis model, lower postoperative mortality and lower incidence of complications in animals as compared with the traditional UUO surgery. The improved UUO group showed statistically significant differences from the sham group in blood biochemical indexes such as urea nitrogen, glutamate aminotransferase, and albumin (P < 0.05). Results from hematoxylin-eosin staining and Masson staining of the obstructed kidney revealed typical pathological features of renal interstitial fibrosis.

Inokosterone effects on proliferation and differentiation of osteoblasts from neonatal sprague-dawley rats / 中国组织工程研究

BACKGROUND: It is an effective strategy for the treatment of osteoporosis to promote the proliferation and differentiation of osteoblasts, but the influence of inokosterone on osteoblast proliferation and differentiation is rarely reported. OBJECTIVE: To investigate the effects of inokosterone on the proliferation and differentiation of primary osteoblasts and the underlying molecular mechanism. METHODS: Primary osteoblasts from neonatal Sprague-Dawley rats were isolated via the second enzyme digestion, and the cells were then cultured in osteogenic induction medium and identified. Cell counting kit 8 assay was performed to evaluate the effect of different concentrations of inokosterone (1, 5, 10 mg/L) on cell viability of primary osteoblasts. Early differentiation ability of osteoblasts was evaluated by alkaline phosphatase staining and alkaline phosphatase activity assay. To evaluate the mineralization ability of osteoblasts, alizarin red staining was performed to observe the number of calcium nodules. The expression level of osteogenic genes was detected by RT-qPCR at different time points. Furthermore, MDC staining was also used to observe the number of autophagosomes. RESULTS AND CONCLUSION: Compared with the control group, inokosterone could inhibit the cell viability of primary osteoblast to some degree (P < 0.05) while significantly promoting alkaline phosphatase activity and mineralized nodules formation (P < 0.05). In addition, inokosterone upregulated the expression of osteogenic genes such as Collagen I, osteoprotegerin, osteopontin, osteocalcin and increased the number of autophagosomes. To conclude, inokosterone can promote osteogenic differentiation by upregulating osteogenic genes expression and activating autophagy of primary osteoblasts.

Reactogenicity of Bordetella pertussis vaccines formulations using aluminum adjuvants in Sprague Dawley rats / Reactogenicidad de formulaciones de vacunas de Bordetella pertussis utilizando adyuvantes de aluminio en ratas Sprague Dawley

Combined vaccines for childhood are a strategy in the prevention of several diseases. These can maximize protection and decrease immunization schedules in children. New candidates are getting closer to being able to meet these needs, but they raise numerous strategic questions related to formulation and regulatory aspects. In addition to being immunogenic and protective must have low reactogenicity when combined with other antigens. Adjuvants are important components in achieving these combinations. Therefore, a reactogenicity study was designed for two Bordetella pertussis formulations containing hydroxide or aluminum phosphate in Sprague Dawley rats. Both formulations dose were administered in 0.2 mL intramuscularly. Clinical evaluations, body weight, water consumption, food, temperature, muscle volume, dermal irritability and pathological studies with special interest at the inoculation site were carried out. Only differences in body temperature and muscle volume were found with a slight increase in values with return to normal. The macroscopic study showed lesions at the site of inoculation, consid­ered characteristics of aluminum adjuvants, such as granulomatous abscesses and the increase in regional lymph nodes near the inoculation site. As conclusion, there are no differences between the formulations of B. pertussis with hydroxide or aluminum phosphate resulted in low reactogenicity.

Las vacunas combinadas resultan una estrategia importante en la obtención de vacunas múltiples para la infancia y el uso de adyuvantes es un componente de gran valor en lograr estas combinaciones, además de ser inmunogénicas y protectoras deben tener baja reactogenicidad, cuando se combinan con diferentes antígenos. Por esta razón, se diseñó un estudio de reactogenicidad a dos formulaciones que contenían hidróxido y fosfato de aluminio con antígenos de Bordetella pertussis en ratas Sprague Dawley. Se administró a cada grupo de ensayo una dosis correspondiente de ambas formulaciones en 0,2 mL por vía intramuscular. Se realizaron observaciones clínicas, comportamiento del peso corporal, consumo de agua, alimentos, temperatura corporal, volumen muscular, irritabilidad dérmica y estudios anatomopatológicos macroscópicos, con especial interés en el sitio de inoculación. No se observaron síntomas, ni muertes en los animales durante el estudio. Tampoco se encontraron diferencias entre los grupos experimentales en cuanto al peso corporal, el consumo de agua y de alimentos; los estudios de temperatura corporal y volumetría muscular evidenciaron un ligero incremento en los valores, los cuales involucionaron rápidamente a la normalidad. En el estudio anatomopatológico macroscópico se observaron lesiones a nivel del punto de inoculación, consideradas propias de los adyuvantes que contienen aluminio, tales como formaciones abscedadas de tipo granulomatosas y el aumento de los ganglios linfáticos regionales cercanos al punto de inoculación. Se concluye que las formulaciones en hidróxido y fosfato de aluminio con antígenos de B.pertussis resultaron ser de baja reactogenicidad.

Effect of Yeonsan Ogye bioactive peptides on anti-oxidant indexes in rats' liver / 한국영양학회지

PURPOSE: This study investigated the effect of bioactive Yeonsan Ogye peptides (YOPs) intake on changes in the hepatic anti-oxidant indexes in male rats. METHODS: Sprague-Dawley male rats were divided into 3 groups and given a casein-based AIN-93G diet and distilled water ad libitum without any added YOPs (control), distilled water with 250 mg of YOPs (Y250), or 500 mg of YOPs (Y500) per kg of body weight for 4 weeks. YOP dose was decided as referred to in the referenced study where toxicity did not occur. The hepatic anti-oxidant indexes were determined using a commercial kit. Statistical analysis was performed using SPSS version 23.0 and are expressed as mean ± standard error of mean. Differences among the groups were evaluated by one-way analysis of variance followed by post hoc Duncan's multiple comparisons test. RESULTS: There were no differences in the body weights, weight gain, food intake, food efficiency ratio, or organ weight, including liver, kidney, spleen, thymus, and epididymal fat, among all of the groups. The hepatic nitric oxide (NO) level in the Y500 group was lower than that in the control and Y250 groups, and the hepatic malondialdehyde (MDA) level was lower in the Y500 group than in the Y250 group. The differences in hepatic superoxide dismutase (SOD) and catalase (CAT) activities were not statistically significant between the groups. From these results we speculated that YOPs may have anti-oxidative abilities to regulate NO and MDA production without affecting SOD and CAT activities. CONCLUSION: YOPs are presumed to act as anti-oxidants in the animal or human body.

Effects of miR-146a on hepatic INSR gene expression in F1 offspring of paternal rats with high-glucose-high-fat diet / 中华内分泌代谢杂志

Objective To study the effect of miR-146a on hepatic INSR gene expression in F1 offspring of paternal rats with high-glucose-high-fat diet ( HGFD) . Methods 5-week-old male SD rats were randomly divided into normal diet ( ND) and HGFD groups. Male rats with ND or HGFD feeding for three months mated with normal female ones. Blood glucose concentration, glucose tolerance, and insulin tolerance in newborn male offspring rats were detected respectively. Differential miRNAs between ND and HGFD groups were compared using next generation sequencing and were then confirmed by real-time quantitative PCR ( qPCR) . The relative expression of INSR mRNA and the methylation level of INSR promoter in liver tissues of the offspring newborn rat were detected and the correlations between them and the relative expression of differential microRNA were analyzed respectively. In vitro, effects of miR-146a on expression and methylation of INSR gene in BRL-3A cells were detected using Western-blot assay and qPCR respectively. Results The fasting glucose concentration of different groups were without significant difference, but glucose tolerance and insulin tolerance of neonatal male rats in HGFD group were decreased significantly . Next generation sequencing has revealed 45 up-regulated miRNAs and 15 down-regulated miRNAs in HGFD group. Among them, differences of 8 miRNAs expression in the enlarged samples were confirmed by qPCR. miR-146a was up-regulated for more than 10 times in the liver of the offspring of HGFD group. Expression level of miRNA-146a was negatively correlated with the relative expression of INSR and positively correlated with the methylation level of INSR in livers of neonatal rats in HGFD group. In vitro, miR-146a ( mimics ) promoted the methylation of INSR gene and inhibited the expression of INSR in BRL-3A cells. Conclusion HGFD feeding to male SD rats leads to the inhibition of hepatic INSR gene expression in neonatal offspring via upregulating miR-146a.

Protein efficiency ratio of pigeon pea (Cajanus cajan) and lima bean (Phaseolus lunatus): A sprague-dawley rat growth assay

@#<p style="text-align: justify;"><strong>OBJECTIVE:</strong> This study has been conducted to determine the protein efficiency ratio (PER) of selected indigenous legumes, namely pigeon pea (Cajanus cajan) and lima bean (Phaseolus lunatus), as alternative protein sources.</p><p style="text-align: justify;"><strong>METHODS:</strong> Experimental diets (pigeon pea-based and lima bean-based) and control diet (casein-based) were formulated and standardized using proximate analysis to have 10% protein basal diet based on the Association of Official Analytical Chemists (AOAC) guidelines. Diets were given to corresponding groups (2 experimental, 1 control) using 30 albino Sprague-Dawley weanling rats aged 21 days old, over a 28-day growth assay.</p><p style="text-align: justify;"><strong>RESULTS:</strong> Results showed that the mean final weight and weight gain were significantly higher in rats fed with casein-based diet (p0.05). The PER of the casein (3.37 ± 2.71) is higher than that of the pigeon pea (1.87) and lima bean (1.32). These results can be attributed to the presence of anti-nutritional factors (ANFs) in the seeds of pigeon pea and lima bean. Reduced protein intake (due to bitter taste), toxicity, and interference with protein digestibility (through interaction with digestive enzymes) imparted by these ANFs can explain the weight loss among the experimental groups and consequently the low PER. Heat treatment done in the study were not adequate to remove the ANFs.</p><p style="text-align: justify;"><strong>CONCLUSION:</strong> The study shows that pigeon pea and lima bean as protein sources alone are not as adequate and efficient in providing necessary protein requirements for weanling rats. Proper processing and treatment should be done to remove inhibitors of protein digestibility and quality.</p>

Effect of intrathecal injection of dexmedetomidine on the behavioral activity, pain degree and expression of protein kinase C in the spinal dorsal horn in rat models of chronic neuropathic pain / 中国比较医学杂志

Objective To analyze the effect of intrathecal injection of dexmedetomidine on the behavioral activity, pain degree and expression of protein kinase C in spinal dorsal horn of rat models of chronic neuropathic pain, and to investigate the analgesic mechanism of dexmedetomidine.Methods 75 healthy male rats were randomly divided into observation group, model group and control group, 25 rats in each group.Chronic sciatic nerve injury model was established in the observation group and model group.After modeling, intrathecal dexmedetomidine intervention was used in the observation group.The model group was treated with saline injection and there was no intervention in the control group.Before the modeling (BM)and at 3(D3), 5(D5), 7(D7), and 14 (D14)days after medicine administration, the behavioral capacity was evaluated by cumulative evaluation method and movement function evaluation, and the assessment of pain degree (mechanical withdrawal method and thermal withdrawal latency pain threshold detection method), PKC staining score (immunohistochemical SABC method), PKC mRNA assay (RT-PCR method) and PKC protein expression (Western blot) were conducted and the data were statistically analyzed.Results ① Before modeling, the behavior, the cumulative scores of motor function, MWT, and TWL showed no significant differences between the different groups (P>0.05).After modeling, the model group and observation group showed that the cumulative scores and motor function scores were increased significantly, MWT and TWL decreased significantly, and the changes in the observation group were significantly lower than those in the model group (P<0.05).After modeling, the cumulative scores, motor function scores, MWT, and TWL were significantly different between the groups (P<0.05).② The expression of PKC was negative in the control group and positive in the model group.In the observation group, after the initial establishment of model, the PKC was strongly positive, and along with the prolonged treatment, the PKC expression intensity was decreased, and only weakly positively expressed at 14 d.③ After modeling, the observation group and model group showed that the PKC mRNA and PKC protein expression levels were significantly higher than that of the control group (P<0.05).With the continuous drug administration, the PKC mRNA and PKC in the observation group were decreasing, and reached a level close to that of the control group at 14 d of drug administration.From the third day after modeling, at the same time points, the amount of PKC expression in the observation group was significantly lower than that in the model group (P<0.05).Conclusions Intrathecal injection of dexmedetomidine can improve the behavior of rat models with chronic neuropathic pain, and reduce the degree of pain.It may be related to the inhibition of protein kinase C expression in the spinal dorsal horn.

Sub-chronic toxicity of MNT-016 by benchmark dose method in Sprague-Dawley rats / 军事医学

Objective To evaluate the repeated dose toxicity of MNT-016 in SD rats and to provide reference for toxicity evaluation.Methods MNT-016 was administered to rats at 5, 20 and 80 mg/kg for 90 days.The toxic effects on the animals were evaluated by observing the clinical signs and measuring the body weight, hematology and blood biochemistry as well as histopathological examination.NOAEL and benchmark dose lower confidence limit(BMDL) were observed by the end point of toxicity.Results Compared with the control group, the AST, TBIL, DBIL and Crea of male rats were increased in a dose-dependent manner, while TG and CHOL decreased.The body mass(before anatomy), heart, liver, thymus, epididymis of male rats in 80 mg/kg group were significantly decreased (P<0.05), while absolute organ mass of the heart and lung was increased.The body mass (before anatomy) and thymus of female rats in 80 mg/kg group were significantly decreased (P<0.05), while absolute organ mass of lungs was increased.Vacuolation of hepatocytes was observed in groups each dose, tubule atrophy was found in the kidneys of 20 and 5 mg/kg groups, and tubule basophilia was observed in 80 mg/kg group.The incidence of the above lesions was higher in male animals than in female ones.Conclusion The NOAEL of MNT-016 is lower than 5 mg/kg in male rats and 5 mg/kg in female rats.BMDL value is 2.65 mg/kg in male animals and more accurate than NOAEL, and is 9.04 mg/kg in female animals,which is slightly larger than the corresponding NOAEL.

Establishment of a modified myocardial ischemia reperfusion model in the SD rat / 医学研究生学报

Objective The establishment of the model of myocardial ischemia-reperfusion injury (MIRI) in SD rats involves lengthy time, severe damage, and death of the rats.We aimed at an efficient and rapid method for establishing an MIRI model in rats to ensure a high success rate of modeling and survival of the animals.Methods Using the trachea intubation-assisted breathing machine, we fully exposed the hearts of 80 adult SD rats (8-10 weeks old and weighing 250-300 mg) by mutilating the 2-3 intercostal muscles.Then, we rapidly positioned and ligated the left anterior descending (LAD) coronary artery for an hour, and established the model of MIRI at 48 hours after releasing the slipknot.We performed electrocardiography (ECG) before, during, and at 1 and 48 hours after operation, ligated the same part again at 48 hours postoperatively, and measured the size of the myocardial ischemia and infarction areas using Evans-TTC double staining.Results The survival rate of the 80 rats was 90% and the success rate of MIRI modeling was 86.25%.After ligation of the LAD coronary artery, ECG showed the ST segment and T wave elevation, followed by gradual decrease of the ST segment and R wave voltage, the myocardium and cardiac apex muscle grey or dark grey below the ligation slipknot.The heart rate myocardial motion amplitude were obviously reduced after ligation.Evans-TTC double staining revealed an evident myocardial infarction area at 48 hours after modeling and the ratio of the ischemia area + infarction area to the total myocardial area of the left ventricle was 0.43 to 0.55 (P>0.05) and that of the infarction area to the ischemia area + infarction area was 0.35 to 0.45 (P>0.05).Conclusion The modified rat model of MIRI can be fast and efficiently established, with a high success rate of modeling and a low mortality of the animals.

Effects of experiment-related factors on hematological parameters in SD rats / 中国比较医学杂志

Objective To study the effects of experiment-related factors on hematological parameters in SD rats, analyze the data difference and causes, understand the effects of anesthetics and stress responses on the physiological aspects of animals, and to provide a reference for the standardization of animal welfare and compound toxicity testing methods.Methods According to gender (A), fasting time (B), anesthesia (C) and blood collection mode (D), SPF SD rats were divided into 24 groups.Blood samples were collected from each group.Then, red blood cell count, hemoglobin levels, white blood cell count and classification indicators were measured.Results The primary and secondary order of the factors affecting the white blood cell count was D > C > A > B, and the levels of white blood cell count of each factor were male rats > female rats, and venous blood > arterial blood, chloral hydrate > pentobarbital sodium > no anesthesia.The primary and secondary order of the factors affecting the white blood cell classification was C > D=A=B, and factors affecting the levels of white blood cell classification were chloral hydrate > pentobarbital sodium > no anesthesia.The primary and secondary order of the effects of the factors on the red blood cell count and hemoglobin level was C > D=A=B, and the levels of red blood cell count and hemoglobin level were pentobarbital sodium > chloral hydrate> no anesthesia.There was no significant difference in the blood indexes between the different fasting time groups.Conclusions There is no effect of fasting on hematological parameters, but there are differences in the blood parameters between arteries and veins.The effect of chloral hydrate anesthesia on the count and classification of white blood cells is greater than that of pentobarbital sodium.The effect of chloral hydrate anesthesia on the red blood cell count and hemoglobin level is greater than that of pentobarbital sodium.The two kinds of anesthesia methods have their own advantages and disadvantages.

Repeated dose toxicity of Tangwang Mingmu granules in Sprague-Dawley rats / 军事医学

Objective To observe toxic symptoms and signs , toxic damage extents and reversibility in rats after oral administration of Tangwang Mingmu granules .Methods Four dose groups with 40 rats in each group were designed in this study, including control group fed with distilled water and three groups at different dosages of the test drug .Tangwang Mingmu granules were orally administered to SD rats at the dosage of 8.4, 4.2 and 2.1 g/kg for 3 weeks and 14.0, 8.4 and 4.2 g/kg for 23 weeks, for 26 consecutive weeks .The general state of the rats was observed every day , while body mass and food consumption were calculated once a week .Halfway through and at the end of the administration (13 and 26 weeks) and after four weeks of recovery, parameters of body mass, hematology, hematological biochemistry, organ/body mass ratio and histopathology were measured .Results Compared with the control group at the same time-point, body mass of male rats in the other three groups was slightly reduced .Food consumption in high and medium dose groups was reduced (P<0.05), MCHC, ALT, TBIL and Na +in high dose group were decreased (P<0.05), TP, ALB and D-BIL were increased (P<0.05), the mean body mass and relative organ weight of thymus in medium dose male rats were decreased (P<0.05), relative organ weight of the liver and kidney in high dose male rats was increased (P<0.05), and focal chronic inflammation to different extent was observed in the liver , kidney and prostate gland .No dose-effect relationship was found in these perturbations that were all within the normal range of animals .No significant drug-related pathological changes were found.Conclusion The NOAEL of Tangwang Mingmu granules is considered to be 14.0 g/kg body mass/day (equal to 50 times the proposed clinical adult dosage ) for the 26-week repeated dose oral toxicity study in male andfemale rats.