Water tank scale: a reliable method for assessing motor function after spinal cord injury in rats / 南方医科大学学报

OBJECTIVE@#To analyze the reliability of the Water Tank Scale for assessing recovery of motor function after spinal cord injury (SCI) in rats.@*METHODS@#Thirty-six adult female SD rats were randomly divided into SCI and sham-operated groups (n= 18). The recovery of the hind limb motor function was assessed using Water Tank scoring, BBB scoring, and motor-evoked potentials (MEP) at 1, 3, 5, 7, 14 and 21 days after SCI. MEP was used as the gold standard for analyzing and comparing differences between the two scoring methods.@*RESULTS@#The Water Tank scores of the rats were significantly higher than the BBB scores on day 3 (0.22±0.43 vs 0, P < 0.05) and also on days 5, 7 and 14 after SCI (0.67±0.49 vs 0.11±0.32, 4.33±1.19 vs 2.83±1.04, 8.61± 1.20 vs 7.06±1.0, P < 0.01). On day 21 after SCI, the scores of the Water Tank Scale of the rats did not significantly differ from the BBB scores (14.78±1.06 vs 14.50±1.47, P>0.05). Neurophysiological monitoring showed that both the Water Tank score and BBB score were significantly correlated with MEP latency, but the Water Tank score had a greater correlation coefficient with MEP latency (r=-0.90).@*CONCLUSION@#Compared with the BBB scale, Water Tank scoring allows more objective and accurate assessment of functional recovery of the spinal cord in early stages following SCI in rats, and can thus be used as a reliable method for assessing functional recovery of the hind limbs in rat models of acute SCI.

Preparation, characterization and biocompatibility of calcium peroxide-loaded polycaprolactone microparticles / 浙江大学学报·医学版

OBJECTIVES@#To explore the physicochemical characteristics and biocompatibility of calcium peroxide (CPO)-loaded polycaprolactone (PCL) microparticle.@*METHODS@#The CPO/PCL particles were prepared. The morphology and elemental distribution of CPO, PCL and CPO/PCL particles were observed with scanning electron microscopy and energy dispersive spectroscopy, respectively. Rat adipose mesenchymal stem cells were isolated and treated with different concentrations (0.10%, 0.25%, 0.50%, 1.00%) of CPO or CPO/PCL particles. The mesenchymal stem cells were cultured in normal media or osteogenic differentiation media under the hypoxia/normoxia conditions, and the amount of released O2 and H2O2 after CPO/PCL treatment were detected. The gene expressions of alkaline phosphatase (ALP), Runt-associated transcription factor 2 (RUNX2), osteopontin (OPN) and osteocalcin (OCN) were detected by realtime RT-PCR. SD rats were subcutaneously injected with 1.00% CPO/PCL particles and the pathological changes and infiltration of immune cells were observed with HE staining and immunohistochemistry at day 7 and day 14 after injection.@*RESULTS@#Scanning electron microscope showed that CPO particles had a polygonal structure, PCL particles were in a small spherical plastic particle state, and CPO/PCL particles had a block-like crystal structure. Energy dispersive spectroscopy revealed that PCL particles showed no calcium mapping, while CPO/PCL particles showed obvious and uniform calcium mapping. The concentrations of O2 and H2O2 released by CPO/PCL particles were lower than those of CPO group, and the oxygen release time was longer. The expressions of Alp, Runx2, Ocn and Opn increased with the higher content of CPO/PCL particles under hypoxia in osteogenic differentiation culture and normal culture, and the induction was more obvious under osteogenic differentiation conditions (all P<0.05). HE staining results showed that the muscle tissue fibers around the injection site were scattered and disorderly distributed, with varying sizes and thicknesses at day 7 after particle injection. Significant vascular congestion, widened gaps, mild interstitial congestion, local edema, inflammatory cell infiltration, and large area vacuolization were observed in some tissues of rats. At day 14 after microparticle injection, the muscle tissue around the injection site and the tissue fibers at the microparticle implantation site were arranged neatly, and the gap size was not thickened, the vascular congestion, local inflammatory cell infiltration, and vacuolization were significantly improved compared with those at day 7. The immunohistochemical staining results showed that the expressions of CD3 and CD68 positive cells significantly increased in the surrounding muscle tissue, and were densely distributed in a large area at day 7 after particle injection. At day 14 of microparticle injection, the numbers of CD3 and CD68 positive cells in peripheral muscle tissue and tissue at the site of particle implantation were lower than those at day 7 (all P<0.01).@*CONCLUSIONS@#CPO/PCL particles have good oxygen release activity, low damage to tissue, and excellent biocompatibility.

Taurine Has no Protective Effect on Rat Corneal Endothelial Cells Injured by Benzalkonium Chloride / 实验动物与比较医学

Objective To observe the protective effect of 2% taurine on corneal endothelial cells injured by benzalkonium chloride in rats. Methods Six piece of corneal endodermis and elastic layer tissue slices were prepared from 6 eyes of 3 SPF SD rats and randomly divided into three groups. The corneal endothelial cells of rats were cultured by tissue block culture for 1 day, then the control group cells were added with 2% taurine solution, while the experimental group cells were added with 2% taurine solution and 0.01% or 0.03% benzalkonium chloride solution. After 1, 2, 4, 5, 6 and 8 days of continuous culture, the growth of corneal endothelial cells in each group was observed under an inverted microscope, and the morphology of endothelial cells was observed under an optical microscope after Wright staining. Results Treated with 0.01% benzalkonium chloride and 2% taurine for 1 day, polygonal endothelial cells appeared on the edge of corneal tissue mass, and the cells were transparent. After 2 days, the number of polygonal cells increased, and there was no fusion growth between cells. After 3 days, the number of polygonal cells decreased and no mitotic signs were observed in endothelial cells. After 4 days, the endothelial nuclei were deeply stained and polygonal cells were rare. After 5 days, the number of endothelial cells decreased, and cell body shrinkage and death occurred. In the experimental group treated with 0.03% benzalammonium chloride and 2% taurine for 1 day, no endothelial cell growth was observed and the cells were sparsely-scattered. In control group, polygonal endothelial cells and a few endothelium-like polygon cells appeared at the edge of tissue blocks after 1 day. After 3 days, the number of polygonal cells at the edge of tissue blocks increased, and there was a phenomenon of gradual fusion growth. After 5 days, the number of endothelial cells increased, and the cells were mostly hexagonal. After 8 days, the endothelial cells formed large sheets, the cell bodies were hexagonal or round, and the nuclei were divided. The growth of corneal endothelial cells in the left and right eyes was uniform, and there was no significant difference in the morphology of the left and right eye endothelial cells in the 0.01% and 0.03% benzalammonium chloride treatment groups and the control group. Conclusion 2% taurine had no protective effect on corneal endothelial cells injured by benzalammonium chloride.

Analysis on Ileum and Colon Microflora of SPF Male SD Rats based on High-throughput Sequencing / 实验动物与比较医学

Objective To analyze the microflora structure and abundance of ileum and colon of SPF male SD rats by high-throughput sequencing technique. MethodsThe ileal and colonic lumen contents of 30 SPF male SD rats were collected, the total bacterial DNA in the contents was extracted and amplified by PCR. The V3-V4 region of bacterial 16S rRNA in the samples was sequenced using Illumina NovaSeq sequencing platform. Then the species structure and abundance of the intestinal flora were analyzed at the phylum and genus level based on validated data. The diversity and differences between ileal and colonic flora were analyzed using the QIIME software (amplicon) analysis tool. The Tax4Fun program was applied to predict the dominant gene enrichment pathways of ileal and colonic flora. ResultsAt the phylum level, the dominant microflora in the ileum of male SD rats were Firmicutes and Actinobacteria, accounting for more than 98%. The dominant colonic microbial community were mainly Firmicutes and Bacteroidetes, accounting for more than 95%. At the genus level, Lactobacillus and Alistipes were the predominant bacteria in the ileum. The colon was dominated by Lactobacillus and Romboutsia. In terms of flora diversity, the alpha diversity of colonic flora including richness index (Chao1 index) and diversity index (Shannon index) were significantly higher than those of ileum (P < 0.01), and the structural variability of the composition of its flora species was smaller than that of ileum; the ileal flora with significant structural variability were mainly of the phylum Firmicutes, Romboutsia, Peptostreptococcaceae, while the colonic flora had significant structural differences in Bacteroidales. In terms of flora function, the ileal flora dominant genes were significantly enriched in lipid-like metabolism, polyketide metabolism, membrane transport, biodegradation and other pathways, while the colonic flora dominant genes were significantly enriched in glycan biosynthesis metabolism, energy metabolism, biosynthesis of cofactor and vitamins and other products, and other pathways. ConclusionThere are significant differences in structure and abundance between the ileal and colonic flora of SPF male SD rats, and the abundance and diversity of colonic flora are higher than those of ileal flora.

Ultrasonic characteristics and histopathologic comparison of vascular remodeling after carotid balloon injury in rats / 中华超声影像学杂志

Objective:To detect the characteristics of vascular remodeling after carotid balloon injury model in rats using ultrasound biomicroscopy(UBM), and to discuss the application value of UBM technique by comparing ultrasonic characteristics with histopathological results.Methods:Carotid balloon injury was performed in 10-week-old SD rats(11 female and 11 male) by 2F Fogarty balloon catheter. The left common carotid artery(CCA) was injured and the right side in the same animal was used as an uninjured control. Arterial structures and hemodynamics were evaluated pre-procedure and post-procedure at 7, 14 days.The intima-media thickness(IMT) inner diameter, outer diameter, lumen area, vessel area, peak systolic velocity, end diastolic velocity of CCA were measured by UBM, and the vascular resistance index, shear stress and blood flow were calculated to evaluate the vascular hemodynamics. The histological data were obtained by H&E staining in cross-sections at 14 days after balloon injury. The characteristics of arterial structure and hemodynamic changes at various time points were compared, the structural changes of CCA between injured and control side after injury were compared. The Spearman correlation and linear regression were used to test the correlation between ultrasonic and histological measurements 14 days after balloon injury.Results:①Compared with pre-procedure, the IMT at 14 days after balloon injury was increased, the inner diameter was decreased, the shear stress in ultrasound was increased(all P<0.05). H&E staining histological test showed that IMT and neointima area in male rats were larger than those of female rats (all P<0.001). ②After carotid balloon injury, the lumen area decreased, but the CCA underwent compensatory positive remodeling and the vessel area increased. ③Significant correlations were demonstrated between UBM and histology in IMT, inner diameter, outer diameter and vessel area of CCA( rs=0.819, 0.965, 0.896, 0.955; all P<0.001). The vessel area value measured by UBM was larger than that of histology( P=0.006). Conclusions:The CCA of rats can be showed clearly by UBM in males and females. The arterial structure cab be measured by UBM accurately with good correlation with histology, as did arterial hemodynamic parameters, which may be benefit for the study in carotid balloon injury model of rats.

Chinese medicine Jiangzhuo mixture regulates glucose and lipid metabolism in obese rats through TLR4/I κB α/NF- κB signaling pathway / 浙江大学学报·医学版

OBJECTIVES@#To explore the mechanism of Chinese medicine Jiangzhuo mixture regulating glucose and lipid metabolism in obese rats.@*METHODS@#Thirty healthy male SD rats were randomly divided into normal control group, model control group, and Jiangzhuo mixture treatment group, with 10 rats in each group. The rats in the normal control group were fed with normal diet, the obesity model was induced by feeding high-fat diet in the model control group and the Jiangzhuo mixture treatment group, the rats in the treatment group were given with Jiangzhuo mixture 50 g/kg by gavage. After 8 weeks of intervention, the blood glucose (GLU), total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) levels were measured in the three groups. Quantitative reverse transcription PCR were used to detect the expression levels of PR domain containing 16 (PRDM16) and uncoupling protein 1 (UCP1) in white and brown adipose tissues of the rats in each group; Western blotting was used to detect the expression of PRDM16 in the white and brown adipose tissue of rats, and Toll-like receptor 4 (TLR4), nuclear factor-κB (NF-κB) and inhibitor of NF-κB alpha (IκBα) in the white adipose tissue; immunohistochemistry was used to detect the expression of UCP1 protein in white and brown adipose tissues.@*RESULTS@#Compared with the normal control group, the white fat weight (P<0.01), white fat coefficient (P<0.05) and Lee's coefficient (P<0.01) were significantly increased in the model control group; the contents of GLU, TC, TG and LDL-C were all increased, and the content of TG was significantly increased (P<0.05) in the model control group. The mRNA and protein expression levels of PRDM16 and UCP1 in white fat and brown fat were significantly decreased (P<0.05) in the model control group. Compared with the model control group, the white fat weight and white fat coefficient and Lee's coefficient were significantly reduced in the Jiangzhuo mixture treatment group (all P<0.01), the levels of GLU, TC, TG, and LDL-C in the the treatment group were all reduced, and the content of TG was reduced more obviously (P<0.01); expression levels of PRDM16 and UCP1 mRNA and protein were increased in brown and white adipose tissue. Compared with the normal control group, the expression levels of TLR4, phospho-IκBα and NF-κB-p65 proteins in white adipose tissue of the model control group were significantly increased (all P<0.01), while the expression levels of these proteins in the treatment group were significantly lower than those in the model control group (all P<0.05).@*CONCLUSIONS@#Jiangzhuo mixture can alleviate high-fat diet-induced increase in body fat, abnormal expression of biochemical indexes and promote the expression of key proteins including UCP1 and PRDM16 in white and brown adipose tissues by regulating TLR4/IκBα/NF-κB signaling pathway.

Improvement in the establishment method for the type Ⅱ cardio-renal syndrome rat model / 中国实验动物学报

Objective To establish an improved type Ⅱ cardio-renal syndrome rat model and evaluate it.Methods Twenty male SD rats were randomly divided into sham and model groups with 7 rats in the sham group and 13 rats in the model group.The model group received the method of squeezing the heart under a small animal anesthesia machine to permanently ligate the left anterior descending branch of the coronary artery to cause myocardial infarction.One week later,unilateral nephrectomy(right nephrectomy)was performed.The rats underwent cardiac echocardiography,pathological staining,and blood and urine tests at 6 weeks to verify model establishment.Results Compared with the sham group,the cardiac function assessed by echocardiography and the endogenous creatinine clearance rate in the model group rats were significantly decreased(P<0.01),and the levels of brain natriuretic peptide,blood creatinine,urea nitrogen,and 24 h urine protein in the model group were significantly increased(P<0.01).HE staining revealed a disordered myocardial arrangement,glomerular atrophy,and inflammatory cell infiltration in model group rats.Picric acid-Sirius red staining showed a significant increase in myocardial collagen fibers,an irregular arrangement of renal tubules,and a large amount of collagen deposition in model group rats.The positive staining area ratio was also significantly increased(P<0.01).Conclusions This improved modeling method provided a typeⅡcardio-renal syndrome rat model with s simple operation,minimal surgical trauma,and low mortality rate.This model simulates the early onset of cardiac and renal function damage and pathological changes in type Ⅱ CRS,laying the foundation for systematic and in-depth research on the pathogenesis and pathological mechanism of type Ⅱ cardio-renal syndrome.

A model study of blood in urine(bladder cancer)during scorching humid fever in MNU-induced SD rats / 中国比较医学杂志

Objective To investigate the modeling process and model establishment rate of the N-methyl-N-nitrosourea(MNU)-induced lower jiao dampness-heat syndrome uremia bladder cancer(BC)model.Methods Batch samples were collected by bladder perfusion(MNU).HE staining and pathological observation of pathological tissues were carried out at six time points before,during,and after modeling to understand the formation and development of bladder carcinoma in situ.Results The experimental process was smooth,and no obvious urinary tract bleeding in rats occurred during the operation.Using this modeling technique,considerable bladder tumors were formed after week 8,and obvious epithelial hyperplasia,damage and large-scale tumor formation were observed in the bladder of rats in the model group.Conclusions MNU induces an SD rat bladder cancer model with obvious model establishment and low mortality.The experimental data provide a reference to establish,improve and apply the bladder cancer model.

Inhibitory effect of ILK-siRNA-AAV on scar formation after glaucoma filtering surgery in rats / 中华实验眼科杂志

Objective:To investigate the role of integrin-linked kinase (ILK)-small interfering RNA (siRNA)-adeno-associated virus (AAV) in scar formation after glaucoma filtering surgery in rat eyes.Methods:Forty-eight Sprague Dawley rats of SPF grade, aged 8 to 9 weeks old, were selected and divided into blank control group, ILK-siRNA-AAV group, NC-siRNA-AAV group and mitomycin C (MMC) group by random number table method, with 12 rats in each group.Left eyes of the rats were taken as experimental eyes, and no intervention was administered to fellow eyes.The bulbar conjunctival filtering bleb after glaucoma filtration surgery in rats was established by anterior chamber drainage tube implantation.One day after operation, phosphate buffer solution, ILK-siRNA-AAV, and NC-siRNA-AAV were injected into the filtering bleb of blank control group, ILK-siRNA-AAV group and NC-siRNA-AAV group, 5 μl each group, respectively.Cotton tablets containing 0.4 mg/ml MMC were placed under conjunctival flap for 5 minutes during operation in MMC group.Intraocular pressure (IOP) was measured with a handheld tonometer before surgery and at 1, 2, 3, 7, 14, 21, 28 days after surgery.Formation of filtering blebs in rats was observed with a surgical microscope at 1, 2, 3, 7, 14, 21, 28 days after operation, and the bleb survival time was calculated by Kaplan-Meier survival analysis.The mRNA and protein expression levels of ILK in conjunctival and subconjunctival tissues at the surgical sites were detected by reverse transcription PCR and Western blot, respectively, on the 28th day after operation.Silencing of ILK gene was identified.Effect of ILK gene silencing on the morphology of drainage pathway was observed by hematoxylin-eosin staining.Effect of ILK gene silencing on collagen fiber deposition in the bulbar conjunctiva at filtration area was examined by Masson staining, and the percentage of positive area of collagen fiber staining in the total tissue visual field was calculated.The use and care of the animals complied with the ARVO Statement.This research protocol was approved by an Ethics Committee of Xi'an Jiaotong University (No.2013-772). Results:There were statistically significant differences in IOP at different time points between before and after surgery among four groups ( Fgroup=76.84, P<0.001; Ftime=114.49, P<0.001). The IOP of ILK-siRNA-AAV group on the 1st, 7th, 14th and 28th day after operation and the IOP of MMC group on the 2nd, 3rd, 7th, 14th and 28th day after operation were lower than those of blank control group, and the differences were statistically significant (all at P<0.05). The IOP of ILK-siRNA-AAV group was lower on 7th, 14th, 21st and 28th day after operation than those of NC-siRNA-AAV group, with statistically significant differences (all at P<0.05). The bleb survival time of blank control group, NC-siRNA-AAV group, ILK-siRNA-AAV group and MMC group was (3.50±1.51), (5.00±3.41), (31.50±3.15) and (31.33±2.46) days, respectively, with a significant difference among them ( F=395.83, P<0.05). The bleb survival time of ILK-siRNA-AAV group and MMC group was higher than that of blank control group and NC-siRNA-AAV group, and the differences were statistically significant (all at P<0.05). There were statistically significant differences in the relative expression levels of ILK mRNA and protein among four groups ( F=222.32, 752.69; both at P<0.05), and the relative expression levels of ILK mRNA and protein were significantly lower in ILK-siRNA-AAV group than blank control group and NC-siRNA-AAV group, and the differences were statistically significant (all at P<0.05). Proliferative fibrous connective tissue and a large number of cells at surgical sites were found in blank control group and NC-siRNA-AAV group, and the fibroblasts were of a high density and grew in clumps.In ILK-siRNA-AAV group, the bulbar conjunctiva was thin, and the arrangement of fibrous connective tissue was loose, and a few proliferative fibroblasts were found.In MMC group, the conjunctival fibrous layer was loose and thin, forming cavities, and scarce cells were found.There was statistically significant difference in the percentage of collagen fiber positive staining area among four groups ( F=741.66, P<0.05). The positive staining percentage of ILK-siRNA-AAV group and MMC group was significantly lower than that of blank control group, among which there was lower positive staining percentage in ILK-siRNA-AAV group than NC-siRNA-AAV group, and the differences were statistically significant (all at P<0.05). Conclusions:Silencing of ILK can inhibit the scar formation after glaucoma filtering surgery and maintain low IOP in rats.

Study on the repair effect of lyophilized platelet lysate products on articular cartilage injury model of rat / 中国输血杂志

【Objective】 To discuss the repair effect of lyophilized platelet lysate (PL) products on articular cartilage injury model of rats. 【Methods】 A total of 25 SD rats were injected with typeⅡcollagenase at the right knuckle articular cavity respectively on day 1, 3 and 5 of experiment, and the modeling conditions were observed 14 days after the last injection of collagenase. The SD rats with successful modeling were randomly divided into three groups, and were injected with lyophilized PL [Group A, 1 mL/(mouse·time)], PL [Group B, 1 mL/(mouse·time)], and normal saline[Group C, 1 mL/(mouse·time)]. The above three substances were injected with corresponding drugs on day 0, 7, 14 and 21 of experiment based on the grouping conditions, and the changes of knee joint diameters of the rats from the three groups were observed and compared. On day 14 and 28, one rat in each group was randomly killed and two knuckle articular cavities of each were taken for tissue sampling, using hematoxylin-eosin staining (HE) and immunohistochemistry. 【Results】 After 14 days of modeling by injection of type Ⅱ collagenase, the proportion of successful modeling in rats was 84% (21/25), with the knee joint diameter (mm) before and after modeling at 12.84±1.14 vs 14.11±1.17(P<0.01). On day 14, 21 and 28, groups A and B were superior to group C in the knee joint diameter and activity improvement (P<0.05), with 13.33±1.16 vs 13.37±1.08 vs 14.21±1.08, 13.10±1.09 vs 13.01±1.04 vs 14.09±1.09 and 12.38±1.08 vs 12.51±1.03 vs 14.01±1.07, respectively. Histological observation showed that group A and B were superior to group C in the production and arrangement of chondrocytes and the positive expression of type Ⅱ collagen, and there was no significant difference between group A and group B. 【Conclusion】 The lyophilized PL has similar therapeutic effect to PL in the treatment and repair of articular cartilage injury, and is worthy of clinical application.

Effect of Buyang Huanwu decoction on expression of silent information regulator 1 in cortical area of rats after cerebral ischemia/reperfusion injury / 中国药理学通报

Aim To investigate the effect of Buyang Huanwu decoction on the expression of silencing regulation factor 1(SIRT1)protein in cortical area and the possible mechanism of cerebral ischemia/reperfusion injury(CIRI)via establishing middle cerebral artery occlusion(MCAO)model. Methods SD rats were randomly divided into sham group(Sham), model group(MCAO/R), Buyang huanwu decoction group(BYHWT), and atorvastatin group(Atorvastatin), with 15 rats in each group. After 2 h ischemia/reperfusion for 72 h and drug intervention, the model was successfully constructed by using laser speckle blood flow monitoring video system. Zea Longa neurological function score was used to evaluate the neurological defects of rats after modeling. TTC staining was used to detect infarct volume. Nissl staining was used to observe the injury of nerve cells. Western blot was employed to detect the SIRT1 protein expression level. Immunofluorescence was applied to detect the fluorescence expression of SIRT1. Results Compared with sham group, the neurological deficits of MCAO/R group were serious(P<0.05). Cerebral infarction volume increased(P<0.05). The nerve cells were severely damaged, disordered, with the nucleus pyknosis(P<0.05). SIRT1 protein expression was reduced(P<0.05). The fluorescence intensity of SIRT1 decreased(P<0.05). Compared with MCAO/R group, the neurological impairment degree of rats in BYHWT and Atorvastatin groups was reduced(P<0.05). The proportion of cerebral infarction volume decreased(P<0.05). The injury of nerve cells was significantly reduced and the number of nerve cells increased(P<0.05). The expression of SIRT1 protein was up-regulated(P<0.05). Fluorescence intensity of SIRT1 increased(P<0.05). Conclusions Buyang huanwu decoction can effectively alleviate brain injury in cerebral ischemia/reperfusion rats, and its protective effect may be related to the increase of SIRT1 protein expression in the ischemic cortical region.

Effect of Da Jianzhongtang on Visceral Pain in Rats with Irritable Bowel Syndrome / 中国实验方剂学杂志

Objective:To investigate the effects of Da Jianzhongtang on substance P （SP）， mast cells （MC）， Toll like receptor 2 （TLR2）， TLR4 on MC model and nuclear transcription factor （NF）-<italic>κ</italic>B p65 in visceral pain rats with irritable bowel syndrome （IBS）， and explore its mechanism of action on IBS visceral pain. Method:Forty-eight 3-day-old SD rats were randomly divided into 6 groups： the control group （control）， irritable bowel syndrome group （IBS）， ketotifen group （Ketotifen，0.18 mg·kg^-1）， Da Jianzhongtang low， medium and high dose groups （DJZT-L， DJZT-M， DJZT-H，2.16，1.08，0.54 g·kg^-1）， with 8 rats in each group. Intragastric administration lasted for 2 weeks. Maternal separation method was used to establish the IBS visceral pain model in rats. The visceral sensitivity of rats was evaluated at 60， 40 and 20 mmHg （1 mmHg≈0.133 kPa） with Abdominal wall withdrawal response （AWR） scale. SP and NF-<italic>κ</italic>B p65 protein expression levels in colon tissue were detected with Western blotting technique. TLR2 and TLR4 proteins on mast cell membrane were detected by immunofluorescence staining. The degranulation rate of mast cells in colon tissue was detected by toluidine blue staining. Result:Compared with normal rats， AWR scores of model rats significantly increased at 60， 40， and 20 mmHg pressure （<italic>P</italic><0.05，<italic>P</italic><0.01）， the degranulation rate of mast cells in colon tissue and SP protein expression in colon tissue significantly increased （<italic>P</italic><0.01）， TLR2， TLR4， and nuclear NF-<italic>κ</italic>B p65 expression on mast cell membrane significantly increased （<italic>P</italic><0.01）. Compared with model rats， the AWR scores of DJZT-H group （pressure of 40， 20 mmHg） and DJZT-M group （pressure of 60， 40， 20 mmHg） significantly decreased. Meanwhile， the degranulation rate of colon mast cells， and the SP， TLR2， TLR4， and NF-<italic>κ</italic>B p65 expression also significantly decreased （<italic>P</italic><0.05，<italic>P</italic><0.01）. Conclusion:Da Jianzhongtang can affect mast cell activity and finally decrease visceral pain of IBS rats by down-regulating SP in colon tissue.

Protective effect and mechanism of growth hormone releasing peptide on diabetic retinopathy in rats / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To investigate the effect of growth hormone releasing peptide(ghrelin)on diabetic retinopathy in rats and study its protective effect.<p>METHODS: Eighteen male SD rats were divided into control group, model group and ghrelin group. HE staining was used to observe the morphology of retina, TUNEL staining was used to observe apoptosis, transmission electron microscope was used to observe the ultrastructure of retinal pigment epithelium, immunohistochemistry was used to detect oxidative stress index, and ELISA was used to detect the content of inflammatory factors. <p>RESULTS:Morphological observation showed that ghrelin could reduce the degree of retinal tissue damage and the apoptosis of retinal tissue in diabetic rats. The results of immunohistochemistry showed that the activities of SOD(superoxide dismutase)and glutathione peroxidase(GSH-Px)in retina tissue of ghrelin group were increased, and the content of malondialdehyde(MDA)was decreased, compared with model group(<i>P</i><0.05). ELISA results showed that intercellular cell adhesion molecule-1(ICAM-1), tumor necrosis factor-a,(TNF-a)and interleukin-1β(IL-1β)in model group were significantly higher than those in control group(<i>P</i><0.05). After ghrelin intervention, the expression of inflammatory factors decreased, compared with model group(<i>P</i><0.05).<p>CONCLUSION:Ghrelin could effectively retard the progression of diabetic retinopathy in diabetic rats, and its mechanism was related to lowering the level of oxidative stress and inhibiting inflammation.

Study on the laboratory historical control values of biological indicators in SD rats with 28-day repeated dose oral toxicity tests / 中国职业医学

OBJECTIVE: To establish the laboratory historical control values for biological indicators in SD rats with 28-day repeated dose oral toxicity tests. METHODS: The body mass, blood routine indexes, serum biochemical indexes, organ mass and organ coefficient of 10 batches of specific pathogen free SD rats in the control group and the control additional group were collected for 28-day repeated dose oral toxicity tests, and the historical control values was established. RESULTS: The body mass of both male and female SD rats increased with the increasing age(all P<0.01). The body mass of male rats was higher than that of female rats each week(all P<0.01). The body mass, blood routine and serum biochemical indexes, organ mass and organ coefficient of SD rats were affected by the age and gender of rats to varying degrees. The effects of age and gender on organ mass and organ coefficient were not consistent. The laboratory historical control values of body mass, blood routine indexes, serum biochemical indexes, organ mass and organ coefficient of SD rats were established according to the age measured in weeks and the gender of rats. CONCLUSION: The laboratory control values of biological indicators of SD rats should be established according to different weekly age and the gender of rats. Organ coefficient is more suitable as an observation index for toxicological safety evaluation compared with organ mass.

Comparison of anxiety susceptibility between FH/Wjd rats and SD rats / 中国药理学通报

Aim With SD rats as control, to observe the anxiety susceptibility of FH/Wjd rats. Methods The anxiety behavior of 3-month-old SD rats and age-matched FH/Wjd rats were evaluated by elevated plus-maze test and open field test. The contents of 5-hydroxytryptamine (5-HT), dopamine (DA) and its metabolite from the cortex were detected by high performance liquid chromatography with electrochemical detection, and the metabolic ratios of DA and 5-HT were calculated. The activities of catechol-O-methyltransferase (COMT) and monoamine oxidase (MAO), the contents of corticosterone (CORT) and adrenocorticotropic hormone (ACTH) were detected by enzymelinked immunosorbent assay in cortex. The expression of COMT was detected by polymerase chain reaction. The expression of all genes hippocampus was detected by mRNA-seq. Results As compared with SD rats, in FH/Wjd rats, the closed arms' distance and the total distance were significandy higher in elevated plus-maze; the central distance was significantly shorter, and the total distance was significantly longer in open field. The contents of DA,5-HT and DOPAC in cortex were significantly lower, and there was no significant difference in HVA and 5-HIAA. The ratio of HVA/DA and 5-HIAA/5-HT, the activity of COMT and MAO, the level of CORT and ACTH, the mRNA expression of COMTwere all higher. The differential genes of FH/Wjd rats and SD rats were mainly enriched in the neuroactive ligand-receptor interaction pathway. Conclusions Compared with SD rats, FH/Wjd rats have lower DA and 5-HT contents, hypermetabolism, hyperactivity of hypothalamic-pituitary-adrenal axis, abnormal expression of COMT and gene encoding neuropsychiatric system. Therefore, FH/Wjd rats have obvious anxiety characteristics.

Background data on embryo-fetal development toxicity studies in SD rats / 中国药理学与毒理学杂志

OBJECTIVE: To establish the background data on SD rat embryo-fetal development toxicity studies by Shanghai Institute of Planned Parenthood Research (National Evaluation Center for Toxicology of Fertility Regulating Drugs), and provide reference for reproductive toxicology research. METHODS: From the embryonic-fetal developmental toxicity tests of SD rats between 2010 and 2018, the index data on embryo development and fetal growth and development of pregnant rats was selected, including pregnancy body mass and pregnancy outcomes (average of corpora lutea, implantation, live fetuses, absorptions and dead fetuses), the growth and development of fetuses (fetal body mass, body length and tail length) and fetal appearance, viscera, and skeleton. The data was statistically analyzed, and the mean, standard deviation and coefficient of variation were calculated. RESULTS: Totally 217 pregnant rats and 2506 fetuses were included. The body mass of pregnant rats on gestation day 0 (GD0) was (222±22) g, while the food consumption of GD0-1was (18.9±3.8) g·d-1. The body mass of GD20 was (353±30) g and the food consumption of GD19-20was (26.2±4.0) g·d-1. GD20euthanasia cesarean section examination showed that the pregnancy rate was 95.2%, and that the average of corpora lutea, implantation and live fetuses was 12.8±2.1, 11.8±2.8 and 11.5±2.8, respectively. The rate of live fetuses and absorption rate was 97.9% and 2.1%, respectively. Among the fetal development indexes, the body mass and placental mass were 3.6±0.3 and (0.5±0.1) g, respectively. The body length was (35.7±1.5) mm, the tail length was (12.9±0.4) mm, and the sexual ratio (male/female) was 1.0 (1227/1279). The external, visceral and skeletal variations were 0.2%, 0.5% and 7.4%, respectively. CONCLUSION: The background data on embryo-fetal toxicity studies of SD rats has been established, which can provide reference for reproductive toxicity research.

Comparative observation on drinking behavior of FH/Wjd rats and SD rats with different liquors / 中国药理学通报

Aim: To observe the preferences and drinking behavior characteristics of congenital alcoholism in (FH/Wjd) rats and SD rats. Methods: Both FH/Wjd rats and SD rats were fed with free access to two bottles (liquor A, liquor B, 53% alcohol and water). The intake of liquor and water and their preference, the difference in day and night intake of each liquor, and the variation of ethanol and water intake and the preference after depriving were compared between FH/Wjd rats and SD rats. Results: The rate of preference for liquor A was significantly higher than liquor B and 53% alcohol in FH/Wjd rats; however, the rate of preference for liquor B was significantly higher than that for liquor A and 53% alcohol, and the liquor A was higher than 53% alcohol in SD rats. It was found that the longer the drinking time, the more of alcohol consumption and the higher preference for each liquor in both FH/Wjd rats and SD rats. In addition, the drinking behavior of FH/Wjd rats and SD rats for each liquor showed a significant difference between day and night, with robust ethanol deprivation effect. Conclusions: SD rats can be forced to drink for short time, and prefer liquor B. However, FH/Wjd rats prefer liquor A. Both SD rats and FH/Wjd rats have the characteristics that the longer the drinking time, the more of alcohol consumption and the higher of preference. Obvious difference exists between day and night; the more alcohol consumption and higher rate of preference after deprivation appear in both SD rats and FH/Wjd rats.

Comparison of valproic acid-induced autism spectrum disorder models between SD rats and C57 mice / 中国药理学与毒理学杂志

OBJECTIVE To compare the difference and similaritiy in growth, neurodevelopment and neurobehavioral characteristics between SD rat and C57 mouse models of valproic acid (VPA)induced autism spectrum disorder (ASD), and select an appropriate experimental species for the study of ASD. METHODS SD rats or C57 mice were ip given VPA 600 mg·kg-1 on embryonic day 12.5 of gestation (VPA groups) or an equal dosage of saline (normal control group). The farrowing rate of pregnant rats or mice and 4-day survival rate were observed. The male pups' malformation rate, body mass, tail length, incisor eruption, eye opening and development of fur were detected to observe physiological development. The neurodevelopment was measured in male pups including the number of days taken by surfacing righting reflex, cliff avoidance reflex, air righting reflex, pivoting, crawling and bar holding test and auditory startle to become positive. In addition, three chamber sociability test, open-field test and self-grooming test were used to assess autistic-like behaviors. RESULTS Compared with C57 mice in normal control group, the VPA groups presented a low farrowing rate in pregnant mice (P<0.05), and high external abnormality in new born offspring (P<0.01). However, there was no significant difference between SD rats in normal control group and those in VPA group. Compared with C57 mice in normal control group, the 4-day survival rate of offspring of the VPA group was significantly reduced (P<0.05), but there was no significant difference between SD rats in normal control group and those in VPA group. The male offspring of C57 mice in VPA group showed significant growth retardation compared with the normal control group, such as lower body mass at 20 and 25 d after birth (P<0.05), delayed incisor eruption (P<0.05) and eye opening (P<0.01). Although the body mass of the male offspring SD rats in VPA group was also significantly lower than that of normal control group at the same time points (P<0.01), there was no significant delay in the incisor eruption or eye opening. Compared with the male offspring of SD rats in normal control group, the VPA group had a significant delay in surface righting reflex (PO.01), cliff avoidance reflex (P<0.01), air righting reflex (P<0.05), crawling (P< 0.01) and bar holding test (P<0.01). Compared with the normal control group, pivoting (P<0.05) and bar holding test (P<0.05) in the C57 mice VPA group were delayed. In the three chamber sociability test, open field test and self-grooming test, the male offspring of SD rats in VPA group showed decreased spatial exploration ability (P<0.01), impaired social preference and novelty preference (P<0.01), and repetitive and stereotyped behaviors (P<0.05). VPA mice only exhibited reductions in spatial exploration ability and novelty preference (P<0.05). CONCLUSION If SD rats and C57 mice are exposed to VPA in the second trimester of pregnancy, their male offspring exhibits behavioral abnormalities relevant to ASD. However, SD rats are superior to C57 mice in building ASD animal models in terms of maternal pregnancy condition, cost and model face validity.

Inhibitory effect of butylphthalide on hippocampal neuron apoptosis in ischemic stroke rats and its influence in p38 MAPK signaling pathway / 吉林大学学报(医学版)

Objective: To investigate the effects of butylphthalide on the hippocampal neuron apoptosis and p38 mitogen-activated protein kinase (MAPK) signaling pathway in the rats with ischemic stroke, and to elucidate the mechanism of butylphthalide in ischemic stroke. Methods: A total of 102 male rats were randomly divided into sham operation group, model group and butylphthalide group; there were 34 rats in each group. The rats in model group and butylphthalide group were used to establish the focal cerebral ischemia models with modified Zea-Longa method. The rats in butylphthalide group were treated with butylphthalide (4. 5 mg · kg-1) after modeling. The rats in sham operation group were intraperitoneally injected with the same volume of normal saline at the same time points. The morphology of neurons was observed by HE staining. The apoptosis of hippocampal neurons was observed by in situ terminal transferase labeling (TUNED staining. The expression levels of activated caspase-3 (cleaved caspase-3), B lymphocyte tumor-2 (Bel-2), Bel-2 related X protein (Bax), p38, phosphorylation-p38 (p-P38) and MAPK proteins in hippocampus tissue of the rats in various groups were determined by Western blotting method. The expression levels of cleaved caspase-3, Bax, Bcl-2, p38, and MAPK mRNA in hippocampus tissue of the rats in various groups were determined by reverse transcription-polymerase chain reaction (RT-PCR). Results: The neurons in the hippocampal CA1 area of the rats in sham operation group were well aligned and the nuclei and cell membrane were normal. In model group, the neurons in the hippocampal CA1 area were disordered, the cells were swollen and burst, and the nuclei had pyknosis. In butylphthalide group, the structure of some neurons in the hippocampal CA1 area returned to normal. Compared with sham operation group, the number of neurons in the hippocampal CA1 area of the rats in model group was significantly reduced (P<0. 01), and the apoptotic index of neurons was significantly increased (P<0. 01). Compared with model group, the number of neurons in hippocampal CA1 area of the rats in butylphthalide group was significantly increased (P<0. 01), and the apoptotic index of neurons was significantly decreased (P<0. 01). Compared with sham operation group, the expression levels of cleaved caspase-3, Bax, p-p38 and MAPK proteins in hippocampal tissue of the rats in model group were significantly increased (P<0. 01), and the expression level of Bcl-2 protein in hippocampal tissue of the rats in model group was significantly decreased (P<0. 01); compared with model group, the expression levels of cleaved caspase-3, Bax, p-p38 and MAPK proteins in hippocampus tissue of the rats in butylphthalide group were significantly decreased (P<0. 01), and the expression level of Bcl-2 protein in hippocampus tissue of the rats in butylphthalide group was significantly increased (P<0. 01). Compared with sham operation group, the expression levels of cleaved caspase-3, Bax and MAPK mRNA in hippocampus tissue of the rats in model group were significantly increased (P<0. 01), and the expression level of Bcl-2 mRNA in hippocampal tissue of the rats in model group was significantly decreased (P<0. 01); compared with model group, the expression levels of cleaved caspase-3, Bax and MAPK mRNA in hippocampus tissue of the rats in butylphthalide group were significantly decreased (P<0. 01), and the expression level of Bcl-2 mRNA in hippocampus tissue of the rats in butylphthalide group was significantly increased (P<0. 01). Conclusion: Butylphthalide can inhibit the apoptosis of hippocampal neurons in the rats with ischemic stroke by inhibiting the p38 MAPK signaling pathway in the hippocampus tissue.

Dynamic changes of pulmonary morphology and mast cell activity in rats after tracheotomy and intubation and their significances / 吉林大学学报(医学版)

Objective: To investigate the dynamic changes of pulmonary morphology and mast cell activity in the rats after tracheotomy and intubation and their significances, and to elucidate the mechanism of occurrence of pulmonary inflammation in the rats after tracheotomy and intubation. Methods: A total of 48 SPF female SD rats were randomly divided into normal control group and tracheotomy and intubation group (n = 2 4) . The lung tissue and alveolar lavage fluid of the rats were collected at 24, 48, 72 and 168 h after modeling, respectively. The pathological changes of lung tissue of the rats were observed by light microscope. Transmission electron microscope was used to observe the mast cell ultrastructures of the rats. The levels of histamine, TNF-a and IL-6 in alveolar lavage fluid of the rats in two groups were measured by ELISA. The expression levels of (3-tryptase in lung tissue of the rats in two groups were measured by immunohistochemical method. Results: The different levels of inflammation of lung tissue of the rats were found in tracheotomy and intubation group under light micscope and the Smith scores were higher than those in control group (P 0 . 05). The levels of TNF-a and IL-6 in alveolar lavage fluid of the rats in tracheotomy and intubation group at different time points were higher than those in normal control group (P < 0 . 05); they reached a peak at 24 h and then declined (P < 0 . 05). The expression levels of mast cell (3-tryptase in the lung tissue of the rats in tracheotomy and intubation group at different time points were higher than those in normal control group (P< 0. 05), and reached a peak at 24 h (P < 0 . 05). Conclusion: The lung mast cell activity is enhanced obviously after tracheotomy and intubation. It may be one of the main mechanisms of complicated pneumonia of the rats with tracheotomy and intubation.