Abstract:Objective To establish a stably overexpressing miR?31 transgenic mouse and detect the expression of miR?31 in the organs and tissues, and to provide qualified tool mice with overexpression of miR-31 in vivo. Methods The miR-31 overexpression vector was constructed by Gateway cloning technology. The vector was injected into fertilized ovum by DNA microinjection technology, then transferred to the pseudopregnant mice and waited for eutocia. Newborn mouse tail DNA was extracted and PCR and agarose gel electrophoresis were performed to identify the positive miR-31 transgenic mice. microRNA was extracted from the organs and tissues of miR-31 transgenic mice and the expression of miR-31 was detected by RT-PCR. The expression of Nestin and number of neural stem cells in the nervous system were compared in the positive and WT mice. Results The miR-31 transgenic mice were constructed successfully and bred more than 14 generations in barrier environment. Expression of miR-31 was increased in major organs and tissues. The expression of Nestin and the number of neural stem cells in the positive mice were higher than those in the wild type mice. Conclusions MiR-31 overexpressing transgenic mice are constructed by Gateway cloning technology and the expression of miR-31 is stable in subsequent generations. The number of neural stem cells in the nervous system is higher than that in wild-type mice. The miR-31 overexpressing transgenic mice can be a good tool for experimental research of the function of overexpressed miR-31 in vivo and the treatment of nervous system diseases.

Abstract:Objective To investigate the effect of sodium houttuyfonate on the expression of PI3K and AKT1 and mTOR mRNA in the lung of rats with chronic obstructive pulmonary disease (COPD), and reveal the possible mechanism of the COPD treated with sodium houttuyfonate. Methods Twenty-four male Wistar rats were randomly divided into normal control group, model control group, dexamethasone group and sodium houttuyfonate group (n =6 for each). The rat models of COPD were established by intratracheal instillation of lipopolysaccharide and smudging. The expressions of PI3K and AKT1 and mTOR mRNA were determined by real-time PCR. The morphological changes of the lung tissue was examined by histopathology. Results Compared with the normal control group, the expressions of PI3K and AKT1 were significantly increased and mTOR mRNA was significantly decreased in the model group ( P <0.01, P <0.05). Compared with the model group, the expressions of PI3K and AKT1 were significantly decreased and mTOR mRNA was significantly increased in the sodium houttuyfonate group and dexamethasone group( P <0.01, P <0.05). Compared with the dexamethasone group,the expression of mTOR mRNA was significantly increased in the sodium houttuyfonate group ( P <0.05). The pathological observation indicated that there were local pulmonary consolidation and a extensive neutrophil infiltration in the alveolar cavity. Prominent pulmonary interstitial fibrous hyperplasia was observed in the model group. The pathological manifestations were much ameliorated than those of the model group, and only mild interstitial pneumonia and a slight fibrous hyperplasia were seen in the sodium houttuyfonate and the dexamethasone groups. Conclusions Sodium houttuyfonate reduces the injury of lung tissue and has protective effect on COPD rats. The mechanism is probably related to the down-regulatation of expression of PI3K and AKT1 mRNA and up-regulatation of expression of mTOR mRNA in COPD rats.

Abstract:Objective To detect the role of PAR2-PKA/ PKCε signaling pathway in periphery neurons in the transition from acute to chronic pain, and investigate the possible approach to prevent both acute and chronic pain simultaneously. Methods SD rats were randomly divided into control group, sham model group, model group, iPAR2-1 group and iPAR2-2 group. The hyperalgesia priming model was established by injection of carrageenan and PGE2 into the left hindpaw except control and sham model group. PGE2 was administrated at 7 days after carrageenan injection. The PAR2 inhibitor was administrated before and after PGE2 injection separately in the iPAR2-1 group and iPAR2-2 group. The paw withdrawal thresholds (PWTs) of rats in each group was detected before and at 5 h, 3 d, 6 d, 7 d 0.5 h, 7 d 4 h, 7 d 24 h after carrageenan injection. The expression level of PAR2, PKA and PKCε proteins in the dorsal root ganglion (DRG) were detected at 24 h after carrageenan injection. Results The hyperalgesia priming model was successfully generated. When PGE2 was administrated at 7 days after carrageenan injection, the hyperalgesia induced by PGE2 was significantly prolonged. The PWTs of rats in the model group were significantly lower than that of the control and sham model groups ( P <0.01), though the PWTs of sham model group had no significant difference with the control on 7 d 24 h after carrageenan injection ( P >0.05). The expression level of PAR2 and PKCε in the ipsilateral DRG neurons were significantly increased on 7 d 24 h after carrageenan injection, when compared with the control and sham model groups ( P <0.05). PAR2 inhibitor prevented the prolonged hyperalgesia induced by PGE2 ( P <0.05) and decreased the PKCε expression in DRG neurons whenever it was given ( P <0.05). However, PAR2 inhibitor did not regulate the acute inflammatory pain of PGE2 and the expression of PKA in DRG neurons ( P >0.05). Conclusions Inhibition of the expression of PAR2 can prevent the transition from acute to chronic pain. This effect may be related with the inhibitory effect on the activation of PAR2-PKCε signaling pathway in DRG neurons. However, inhibition of PAR2 can not regulate the acute pain. These may because of that the PAR2-PKA signaling pathway does not play a role in acute pain.

Abstract:Objective To observe the changes of renal tubular injury and the extent of interstitial fibrosis in the C57BL/6 mouse models of chronic kidney disease (CKD), and provide experimental animal evidence for study of the progression of acute kidney injury (AKI) to chronic kidney disease as well as its mechanisms. Methods Twenty-four 8 week-old male C57BL/6 mice were randomly and equally divided into control group, low-dose, medium-dose, and high-dose cisplatin groups, 6 mice in each group. Mice in the cisplatin groups were administrated with 5, 7 or 10 mg/ kg cisplatin by intraperitoneal injection once a week for 4 weeks. Plasma creatinine and 24-hour urinary protein were detected to assess the renal function. The mice were sacrificed, and plasma and kidney samples were collected for subsequent tests. Pathological changes were observed using periodic acid-Schiff (PAS) staining. To evaluate renal tubules injury, the expression of kidney injury molecule 1 (KIM-1) was examined by immunohistochemistry and the level of urinary N-Acetyl-β-D-glucosaminidase was detected with a commercial kit. The infiltration of CD3-positive T cells and F4/80-positive macrophages was observed by immunohistochemistry (IHC) and immunofluorescence. The expression of collagen I and α-smooth muscle actin (α-SMA) were tested by immunohistochemistry to assess the renal fibrosis, while total kidney collagen was detected by Picrosirius red staining. Results In contrast to the normal control group, the kidney injury became more serious in the cisplatin-treated mice as cisplatin concentration increased. Particularly, significant kidney damage was observed in the high-dose cisplatin group. Compared with the control group, the plasma creatinine and 24-hour urinary protein were significantly increased in the high-dose cisplatin group ( P <0.05 and P <0.001) indicating impaired renal function. Morphologically, numerous clear vacuoles and necrosis were present in renal tubule epithelial cells in the high-dose cisplatin group. The expression of KIM-1 was markedly up-regulated and the level of urinary NAG was elevated. Infiltration of CD3-positive T cells and F4/80-positive macrophages was enhanced in the mice of high-dose cisplatin group. Data from immunohistochemistry and picrosirius red staining showed that mice of the high-dose cisplatin group developed renal fibrosis evidenced by markedly up-regulated expression of collagen I and α-SMA. Conclusions Repeated administration of 10 mg /kg cisplatin for 4 weeks can induce chronic renal insufficiency in mice, which may serve as a novel model for the research on underlying mechanisms of progression from acute kidney injury to chronic kidney disease.

Abstract: Objective To evaluate the therapeutic effect of chemotherapeutic drugs on pancreatic carcinoma based on patient-derived xenograft (PDX) models, and to screen an individualized treatment strategy. Methods Fresh human pancreatic carcinoma tissues were subcutaneously transplanted into nude mice to establish PDX models which could be stably passaged. The traceability of PDX models was determined by STR analysis. The PDX models were treated with three different clinical chemotherapeutic drugs oxaliplatin, gemcitabine and irinotecan, respectively, and the tumor volumes were measured at different times. The therapeutic effect of those drugs was assessed by TGD mathematical model and plasma CA19 -9 test. Results The traceability of patient-derived xenograft samples was up to 99. 99%. Compared with the control group, the treatment with irinotecan and gemcitabine inhibited tumor growth significantly ( P = 0.001), and gemcitabine had even better result . The minimum toxic effect in the mice was induced by irinotecan treatment, followed by gemcitabine treatment. Conclusions Pancreatic carcinoma PDX models are successfully established and can be stably passaged. Gemcitabine shows the most inhibitory effect on tumor growth based on TGD mathematical model assessment, and deserves to be recommended as the preferred drug for individual treatment of pancreatic carcinoma.

Abstract:Objective To establish a rat model of glucocorticoid?induced osteoporosis (GIOP) and to explore the interventional effect of the Chinese medicine Fufang Zhenzhu Tiaozhi (FTZ) capsules on regulation of mitogen?activated protein kinase kinase kinase 2 (MEKK2)?Wnt coupling and inhibiting β?catenin ubiquitination, and to investigate the effect of FTZ on the bone mineral density and cell osteogenic ability. Methods SPF male rats were randomly divided into normal control group, methylprednisolone group (model group), methylprednisolone + saline group (blank control group) and methylprednisolone + FTZ group (experimental group). The proximal femoral cancellous bone was examined by micro?CT and histopathology, and assessment of expressions of Wnt3a, MEKK2, and β?catenin proteins. Bone mesenchymal stem cells (BMSCs were isolated and treated with serum containing FTZ, stained by alkaline phosphatase and alizarin red. The expressions of osteogenic differentiation?related genes ALP, Runx2 and OCN, the expressions of MEKK2 and β?catenin proteins, and the transcription level of β?catenin/ TCF were determined. Results 1) The micro?CT imaging showed that compared with the control group, the BV/ TV, Tb. Th and Tb/ N expressions were significntly decreased, and Tb/ sp increased in the experimental group ( P < 0.05). Region of interest (ROI) three?dimensional reconstruction of trabecular bone in the experimental group showed improvement of bone trabeculae and local bone repair. 2) The pathology using hematoxylin and eosin staining showed that in the experimental group, the bone trabecular density was higher than that of the model group, and observed a better trabecula morphology. 3) The Wnt3a, MEKK2 and β?catenin expressions in the experimental group were significantly increased compared with the model model ( P < 0.05). 4) After treated with FTZ and BMP2, the result of alkaline phosphatase and alizarin red staining indicated an enhanced osteogenic response ( P <0.05) in the GIOP rat models. 5) After treatment with seum containing FTZ, The BMSCs isolated from the GIOP rats enhanced the transcriptional activity of β?catenin/ TCF/ LEF ( P <0.05) and promoted the expression of β?catenin and MEKK2 proteins ( P <0.05). Conclusions FTZ can ameliorate GIOP by regulating the MEKK2?Wnt coupling and inhibiting the β?catenin ubiquitination, and improve the bone microstructure.

Abstract:Objective This study was conducted to establish a stable and highly efficient method for isolation and purification of pancreatic islets from NOD mice and to evaluate their characteristics in vitro and in vivo. Methods The islets were isolated from mouse pancreas using modified collagenase digestion and Ficoll density gradient centrifugation. The endocrine secretory function was assessed by insulin secretion in either low or high dose glucose stimulation. To evaluate the function of the graft, body weight and blood glucose were monitored, and IVGTT was performed. In addition, to assess survival of the implanted islets, Pathology using HE staining and insulin immunostaining of the graft were performed. Results The average islet yield was 116 ±12 islets/ pancreas and purity was higher than 90%. Compared with islets from Kunming mice, the islets isolated from NOD mice were poorly responsive to glucose challenge. Blood glucose levels and body weight changes of the islet-transplanted diabetic mice were significantly improved compared with the sham-operated mice. In addition, blood glucose levels in vivo after an IVGTT also significantly improved. However, these improvements were only main-tained for 2 weeks. Furthermore, HE staining and immunostaining assays demonstrated that there were insulin-positive cell clusters and lymphocyte infiltration in the graft-bearing kidney. Conclusions A large number of quality islets can be isolated and purified from NOD mice by using the modified mouse islet isolation method, which can be used to develop therapeutic strategies to protect transplanted islets from rejection and autoimmune attack.

Abstract:Objective To investigate the efficacy and safety of the affinity adsorption material for removal of intestinal endotoxin in the blood. Metheds To establish a canine model of intestine-derived endotoxemia by ligation and perforation of the appendix. Hemoperfusion was performed to treat the endotoxemia in model dogs. In the experiment, we monitored vital signs and detect the content of endotoxin and blood physiological and biochemical indexes at different time points. Results The endotoxin content was (0.49 ± 0.22), (0.034 ± 0.00) Eu/ mL after hemoperfusion for 1 and 2 hours, compared with that before the beginning of perfusion(7.25 ±1.18)Eu/ mL, showing a significant difference ( P <0.05). After treatment for 2 hours, the average clearance rate was 99.52%. White blood cells, red blood cells, hemoglobin, platelets, total protein, albumin, globulin, alanine aminotransferase, aspartate aminotransferase, urea and creatinine index were significantly decreased after perfusion ( P < 0.05). The index values were in a normal range and vital signs were stable during the perfusion. Conclusions The affinity adsorption material developed by our team can effectively remove endotoxin in the blood of experimental dogs and has good blood compatibility.

Abstract:Objective Cysteinyl leukotrienes are potent inflammatory mediators. Their actions are mediated by specific receptors, the CysLT receptors (CysLT1 R and CysLT2 R), which have been cloned and characterized. In this study, we investigated the protective effects of the CysLTR antagonist Pranlukast and HAMI 3379 on global cerebral ischemia/reperfusion (CI/ R) injury in gerbils and its underlying mechanisms. Methods The gerbil model of CI/ R was established by bilateral common carotid artery occlusion for 10 min followed by 24 h reperfusion. Then the animals were equally randomized into four groups: sham, model, Pranlukast (0.1 mg/ kg) and HAMI 3379 (0.1 mg/ kg)groups. The later two groups were treated with intraperitoneal injection of Pranlukast and HAMI 3379,respectively,once daily for 4 days before carotid artery occlusion, while the former two groups with saline only, all at 10 mL/ kg. After 24 h reperfusion, neurological deficit scores were observed and the behavioral dysfunction was assessed. The neuron morphology of cerebral cortex and CA1 subregion of hippocampus were observed in brain sections stained with cresyl violet. The expression of autophagy-related proteins beclin-1 and LC3 in the homogenate of cerebral cortex and hippocampus were determined using western blottingm analysis. The ultrastructure of autophagosomes in the CA1 subregion of hippocampus was observed by electron microscopy. Results Compared with the model group, Pranlukast and HAMI 3379 attenuated neurological deficits, improved the behavioral dysfunction, inhibited the neuron injury and loss, decreased the expression of autophagy-related protein beclin-1 and LC3 and the number of autophagosomes. Conclusions cysteinyl Leukotriene receptor antagonist Pranlukast and HAMI 3379 can alleviate global cerebral ischemia/ reperfusion injury in gerbils. The protective effects of Pranlukast and HAMI3379 appear to be associated with the inhibition of autophagy.

Abstract:Objective The aim of this study was to establish a PM2.5 air pollution-induced mouse model of pulmonary inflammation and investigate its pathogenetic mechanism. Methods 150 specific pathogen-free BALB/ c mice were subjected to intratracheal instillation of 2.5, 5, or 10 mg/ kg PM2.5 suspension to construct airborne inflammation models. The blank group and saline group were taken as a control group. Mice were euthanized after 3rd, 7th, 21st, 35th and 49th days to assess the pathological changes in lung tissues using HE staining and ELISA. Results The success rate of tracheal instillation was 96%. With the time prolongation and increasing doses of intratracheal PM2.5 instillation, the histopathological scores of lung tissue increased gradually, showing alveolar macrophages with engulfed particles and lymphocyte accumulation in bronchiole and widened inter-alveolar space. The levels of BALF IL-6 and TNF-α of lung tissue homogenate were significantly increased in the high dose PM2.5 (10 mg/ kg) group, compared with the control groups. Conclusions A mouse model of PM2.5 air pollution-induced lung inflammation is successfully established by intratracheal instillation of PM2.5 suspension. This method is proved to be simple, safe and reliable, and is useful for further study of air pollution-induced and other inflammatory mechanisms.

Abstract:Objective The aim of this study is to explore the changes of expression of integrin αvβ3 in bovine uterine epithelial cells in vitro induced by estrogen or/ and progesterone alone or in combination, and to provide a new reference marker for determining bovine uterine receptivity state. Methods RT-PCR was used to analyze the transcriptional changes of αvβ3 expression in bovine endometrium treated by different concentrations of estrogen, progesterone alone or in combination. Results The expression of integrin αvβ3 reached the highest level when the culture medium was added with progesterone at the concentration of 10 ^-7 mg/ mL, and the expression of αv and β3 in the 10 ^-7 mg/ mL concentration group was significantly higher than that of the control one ( P <0.05). Moreover, the expression of αv was highest in the 10 ^-10 mg/ mL E2 group, but the expression of β3 was the lowest in that one. In addition, adding with both estrogen and progesterone, the transcriptional level of integrin αvβ3 was significantly higher than that in the control one. The transcriptional level of αv in the treatment group was significantly higher than that in the control group ( P <0.05), but the transcriptional level of β3 in this group was not ( P >0.05). Conclusions It can be concluded that integrin αvβ3 can be used as a new potential reference marker gene for detecting the bovine uterine receptive status.

Abstract:Objective To establish a monkey model of muscle atrophy by fixing the right lower limb and set up evaluation criteria for that model. Methods Twelve healthy, 6 -7 year old male cynomolgus monkeys (body weight 6 -7 kg) were used in this study. All animals were normal and had no malformed lower limb bones. Right lower limbs of the animals were fixed with fiberglass bandage(from knee joint to anklebone) for 15 weeks. Body weight, crus perimeter and crus volume were recorded every two weeks. MRI examinations of the gastrocnemius muscle were conducted at weeks 11, 13 and 15. Muscle biopsy was taken for pathological examination at week 15. Results There were no obvious changes of body weight during the experiment. At 15 weeks after modeling, the right crus perimeter and crus volume of the experimental animals were significantly decreased ( P <0.05), and some biochemical indexes such as serum creatinine, gloucose and alkaline phosphatase were significantly decreased ( P < 0.05), while cholesterol was significantly increased ( P < 0.05). MRI showed atrophy of the right gastrocnemius muscle. Muscle biopsy also showed muscle atrophy and muscle fibers became thinner in the right gastrocnemius, and the cross-section area of the muscle fibers was significantly decreased. Conclusions It is easy to operate and establish a muscle atrophy model in Cynomolgus monkeys by fixing the right lower limb. This model is suitable for objective and easy evaluation of muscle atrophy by measurement of crus perimeter and crus volumen of the limb, blood biochemical parameters, MRI examination, and histopathological examination in combination.

Abstract:Objective To observe the dopamine (DA) concentrations in the lateral geniculate nucleus in guineab pigs with flickering light-induced and form deprivation myopia, and to compare and analyze the pathogenetic mechanisms in the centers of vision of these two different myopia models. Methods Twenty-four two-week-old guinea pigs were randomly assigned to three groups (n =8): flickering light-induced myopia (FLM) group, form deprivation myopia (FDM) group and control group. All the groups were fed for 8 weeks. The refraction and axial length (AL) were measured before and after modeling. After eight-week modeling, the contents of dopamine in the left lateral geniculate nucleus were detected by high performance liquid chromatography with electrochemical detection (HPLC-ECD). Results Before modeling, no significant difference was found in refraction and AL among the three groups. After eight-week modeling, in contrast with the control group, significant differences were found in changes of both refraction ( P <0.001) and AL ( P <0.05) in the right eyes of FLM and FDM groups, indicating that the two myopic models were successfully established. The result of HPLC-ECD showed that the contents of DA in the left lateral geniculate nucleus in FLM group were significantly higher than the control group ( P =0.01), while in the FDM group it was lower than the control group ( P =0.021). The average contents of DA were as follows: (37.04 ± 1.18) pg/ μL in the control group; (24.27 ± 3.46) pg/ μL in the FDM group; and (45.58 ±1.98)pg/ μL in the FLM group. Conclusions The content of DA in the lateral geniculate nucleus is increased in FLM, while decreased in FDM, indicating that the expression of DA in LGN and the mechanisms of formation of these two experimental myopia are different.

Abstract:Objective To establish a rat xenograft model of human uterine leiomyoma using immunosuppressive agent and provide a useful tool for the study on uterine leiomyoma. Methods Intragastric administration with immunosuppressive agent mycophenolate mofetil (MMF) (40 mg/ kg) was given to rats for two weeks before the surgery until the end of the experiment. 20 SPF female Wistar rats were randomly divided into 4 groups after abdominal transplantation of human leiomyoma tissues: group A received femoston containing 0.4 mg/ kg estradiol and 2 mg/ kg dydrogesterone, group B received estadiol 0.4 mg/ kg, group C received dydrogesterone 2 mg/ kg, and group D served as the control group, received distilled water 1 mL/200 g. All rat received the corresponding drugs once per day for 2 days. Samples were taken at 4 weeks after the surgery to observe the pathology of the tumor tissues. Results The modeling success rates were 90% in the group A, 40 % in the group B, and 0% in the groups C and D. Conclusions Rat xenograft model of human leiomyoma can be successfully established using an immunosuppressive agent femostone with a high modeling success rate and low cost. It can be used as a new animal model for the study of transplanted leiomyoma.

Abstract:Objective To investigate the structural and functional changes of immune system in aging Sprague-Dawley rats. Methods Sixty SPF 4 -6-week old SD rats (male∶ female =1∶ 1) were used in this study. Ten of them were randomly taken and euthanized every 6 months. The dynamic changes of T cell proliferation, expression of cytokines, serum levels of SOD and MDA and histopathology were examined. Results Obvious histological changes of thymus and spleen were observed in the aging rats. Compared with the young SD rats, in the aging rats, the lymphocyte transformation ability was decreased ( P <0.05, P <0.01), number of splenic cells was declined ( P <0.05, P <0.01), NK ( P <0.05, P <0.01) and T cell subset was reduced ( P <0.05, P <0.01), and the production of IL-2 was decreased as well ( P <0.05, P <0.01). The serum level of SOD in old rats was lower, MDA was increased, with significant differences ( P <0.05, P <0.01). The immunohistochemical staining showed that more extensive staining was found in the nuclei of thymocytes from the aging rats, while the 8?OHdG formation in thymic tissues was mostly located in the thymic medulla. Conclusions Aging process is accompanied by immune impairment, oxidative stress can also impair the immune response in aging rats. Our findings indicate that structural and functional alterations of immune system in aging rats may be closely related with oxidative damages.

Abstract:Objective To establish a rat model of spinal root avulsion and to validate the model by brain-derived neurotrophic factor (BDNF) treatment. Methods To evaluate the motor neuron loss, 5 male SD rats were used to undergo spinal root avulsion surgery. One week later, the number of motor neurons in the ventral horn of the spinal cord was assessed by histopathology using immunohistochemical staining with a choline acetyl transferase (ChAT) antibody. After this pilot study, 40 male SD rats at 7 weeks of age were randomly divided into 4 groups: two control groups, BDNF preventive and treatment groups. Results All rats recovered well post-surgery and no obvious abnormality was observed. Compared with the contralateral side, the number of motor neurons in the ipsilateral avulsed side was significantly decreased at one week after surgery (20.06%, P <0.05). Compared with the control group, there was a significant increase in ChAT positive neurons in the BDNF preventive group (17.85% vs. 93.06%, P < 0.0001) or BDNF treatment group (1week after surgery) (26.94% vs. 86.87%, P < 0.0001), indicating that the motor neurons were effectively protected by BDNF. Conclusions A rat model of spinal root avulsion is successfully established, which can be valuable for studies of amyotrophic lateral sclerosis and drug discovery efforts.

Abstract:Objective To investigate the effect of a Chinese medicine formula GAPT, a combination containing Chinese herbs ginseng, Acorus tatarinowii, Polygala and tuber curcuma, on the behavior and cholinergic system in mice with scopolamine-induced memory impairment, and to explore the neuroprotective mechanism of GAPT. Methods ICR mice were randomly divided into normal control group (solution of 0.5% carboxy methyl cellulose, CMC), model group (0.5% CMC), positive control group (donepezil, 0.92 mg/ (kg·d)), GAPT high, medium and low dose groups (20 mg/ (kg·d), 10 mg/ (kg·d), 5 mg/ (kg·d)), 18 mice in each group, were given intragastric gavage once a day for 30 days. After the last administration, the control group and model group received intraperitoneal injection of normal saline, and the other groups intraperitoneal injection of scopolamine (3 mg/ (kg·d)), dissolved in 0.9% normal saline. Morris water maze test was performed to assess the learning and memmory ability. Then the mice were killed and the Ach content and AchE and ChAT activity in the cortex and hippocampus were detected. Results GAPT increased the swimming distance, swimming time and the residence time in target quadrant of the model mice, increased the content of Ach and the activity of ChAT, and decreased the activity of AchE in the brain of the model group. Conclusions GAPT can improve the learning and memory ability of mice induced by scopolamine, and its mechanism may be related to cholinergic system.

Abstract:Objective To investigate the effects of different high?fat diet feeding time durations on blood glucose (BG), insulin resistance index (HOMA-IR), and urinary albumin excretion rate (UAER) in rats with high fat diet induced type 2 diabetic nephropathy (DN). Methods Unilateral renal artery ligation, high-fat diet (throughout the experiment period), and low dose streptozotocin (STZ) intraperitoneal injection were used to establish a type 2 DN rat model. After the operation, rats in the DN1 and DN2 groups received an intraperitoneal injection of STZ 30 mg/ kg after 4 and 8 weeks of high-fat diet feeding, respectively. UAERs of the DN1 and DN2 groups were compared at 4 weeks after the STZ injection and the end of study (EOS). BG, body weight, HOMA-IR, kidney index, and pathological changes of the kidney were observed. Results UAER was increased in both groups at 4 weeks after the STZ injection, but significantly higher in the group DN2 than in the DN1 group ( P <0.01). At the end of study (the 12th week), the renal tissues showed pathological changes, including glomerular capillary loop hypertrophy, increased mesangial matrix, and decreased capsule space in both groups. Compared with the DN1 group, the body weight was significantly higher ( P <0.01), kidney index was significantly lower ( P <0.01), while BG, serum insulin level, HOMA-IR, and UAER were no significantly changed in the DM2 group ( P >0.05, respectively). Conclusions The results show that extending the feeding time of high-fat diet can aggravate the kidney damages in diabetic rats, but it can also delay the start of any planned intervention. Therefore, the experimental protocol should be carefully designed based on the study objective .

Abstract:Objective To investigate the effects of Qingzao Jiufei Tang (QJD) and its decomposing agent on the levels of TNF-α, INF-γ levels of TNF- α of Qingo, P1 and AQP5 in lung tissue of mice infected with Mycoplasma pneumoniae (MP), and to clarify its molecular anti-infective mechanism. Methods One hundred-forty-four SPF grade BABL/c mice were randomly divided into the normal group (group A), model group (group B), QJD group (group C), QJD decomposition agent I group (group D)?QJD decomposition group II Group (group E) and azithromycin group (group F), with 24 rats in each group. Besides the group A, the other 5 groups of mice were treated with MP infection. After the modeling, the mice were given corresponding drugs by gastric gavage, and samples were obtained on the days 3, 7, 10, 14 after the model was established. The lung tissue sections were examined by histopathology, and the degrees of inflammation in lung tissues in the mice were evaluated, and the lung index and the ratio of dry and wet lung weight ratio in the mice were calculated. The levels of MPN372 and P1 genes were determined by qPCR assay. The serum TNF-α and INF-γ levels in the mice were assessed by enzyme linked immunosorbent assay (ELISA). The expression of AQP5 protein was detected by Westernblot. Results After MP infection, the pathological examination revealed thickening of alveolar septum and bronchioles, and extensive inflammatory cell infiltration in the lung tissues. The lung index was increased and the ratio of dry and wet lung weight ( P < 0.05). TNF- α and INF-γ cedd and was increased, and reached the peak on the seventh day. The expression of AQP5 protein showed a downward tendency, and began to gradually increase on the 14th day. Compared with the group B, the expression levels of MPN372, P1 and TNF-α in the group D were down-regulated, and the expression levels of INF-γ levein the group E were up-regulated from the 7th day. Conclusions QJD can control pulmonary inflammation in mice after MP infection. The decrease of production of MP toxin MPN372 and the expression of adhesion protein P1, the up-regulation of expression of INF-γ and AQP5 proteins, and down-regulation of TNF-α expression are one of the mechanisms of its action.

Abstract:Diabetic foot with diabetic peripheral neuropathy (DPN) can lead to peripheral nerve dysfunction. Exploring appropriate animal models of the disease can make a better understanding of the occurrence, development, pathogenesis, prevention and treatment of DPN. Therefore, a reasonable construction of animal models is related to the success of experimental research. Up to date, experimental animals used in the research of diabetic foot neuropathy include rats, nude mice, transgenic mice, and so on. Nerve injury models can be generated by different means, such as ischemia reperfusion, resection or ligation of nerves and scalds. This article will review the recent research progress of diabetic foot neuropathy animal models.

Abstract:Postpartum depression (PPD) is one of the most common types of postpartum psychiatric syndromes. Because of the complex and changeable characteristics in PPD disease and the special period after childbirth, there are many clinical limitations in the study of this disease. Therefore, the preparation and establishment of a proper animal model closed to clinical and behavioral evaluation method plays an important role in study of its pathogenesis. This review mainly introduces the commonly used postpartum depression animal models and the behavioral evaluation method. It is hoped to provide a reference for further study of PPD pathogenesis and for the drug research and development.