Abstract:Objective To investigate the effects of melittin on Th17/ Treg balance and arthritis-associated inflammation in a rat model of collagen-induced arthritis. Methods Thirty Sprague Dawley rats were randomly allocated into a normal group, while 24 SD rats were injected with bovine collagen II on the 1st day and again on the 10th day to establish a collagen-induced arthritis model. On the 14th day, elimination of 6 unmodeled rats and 18 model rats in which the model had been successfully established were selected and divided into a model group, a melittin group, and a methotrexate group by a random number table method. On the same day, the melittin group was injected with melittin (0.5mg/ kg, once every other day), while the methotrexate group was injected with methotrexate (1 mg only, once per week,three times). The model group and the normal group were intraperitoneally injected with equal amounts of distilled water;then, on the 32nd day, the rats were executed. The concentrations of TNF-α, IL-10, and IL-17 A in serum were determined by ELISA. The spleen of each rat was removed, the leukomonocytes were extracted, the activity of leukomonocytes was detected by MTS, and the proportions of Th17 and Treg cells were determined by flow cytometry.Pathological changes of the ankle joint were also observed by HE staining. Results Melittin and methotrex significantly decreased the concentrations of TNF-α and IL-17 A ( P < 0. 01, P < 0. 05), increased the concentration of IL-10 ( P <0.05), inhibited the activity of leukomonocytes ( P < 0. 05), decreased the proportion of Th17 cells ( P < 0.05), and increased the proportion of Treg cells ( P < 0. 01, P <0. 05). In the melittin group and methotrex group, the proliferation of synovial membrane was decreased, which prevented formation of the pannus and delayed the destruction of bone and articular cartilage. Conclusions Melittin can regulate Th17/ Treg balance and inhibit joint inflammation in rats with collagen-induced arthritis.

Abstract:Objective To investigate the pharmacokinetic parameters of oseltamivir and the efficacy of oseltamivir against influenza virus in tree shrews. Methods Tree shrews were orally administered oseltamivir for pharmacokinetic analysis, and then their pharmacokinetic parameters were compared with those of humans, ferrets, mice, and rats reported in the literature. The tree shrews were inoculated with human influenza virus A/ California/04/2009 H1N1 or avian influenza virus H9N2 Y280-PB2-E627K intranasally. Oseltamivir was orally administered twice a day for 5 consecutive days. Nasal symptoms of the tree shrews were observed, and the viral titer of nasal lavage was determined on 1, 3, and 5days after inoculation, the cell sedimentation of nasal lavage fluid was classified and counted. After 21 days of infection, the tree shrews were executed humanely and the serum antibody titers were determined by hemagglutination inhibition test.Results The pharmacokinetics of oral oseltamivir in tree shrews was as follows: C_max 1. 34 μg/ mL, T_max 0. 75 h, T_{1/ 2}2. 03h, and AUC_0-12 1. 76 mg·h/ liter. The viral shedding of tree shrews infected with H9N2 Y280-PB2-E627K virus was clearly inhibited after 1 day in the high-dosage oseltamivir treatment group, however, there was no clear inhibitory effect for A/California/04/2009 H1N1 virus. High-dosage oseltamivir treatment also reduced the nasal lavage fluid cell count of tree shrews infected with H9N2 Y280-PB2-E627K virus. The antibody titer of tree shrews infected with A/ California/04/2009 H1N1 virus was significantly higher than that of H9N2 Y280-PB2-E627K avian influenza virus. Conclusions The pharmacokinetic parameters of oseltamivir in tree shrews differ from those in humans and other model animals, such as ferrets, however, the T_max and T_{1/ 2}of oseltamivir in tree shrews were similar to those of mice. Oseltamivir can effectively inhibit the shedding of H9N2 Y280-PB2-E627K viruses and alleviate the inflammatory reaction in tree shrew nasal cavity.

Abstract:Objective To explore the feasibility of establishing a tree shrew model of Human Noroviruses (HuNoVs) infection by inoculation with HuNoVs. Methods Three groups of 6-month-old tree shrews (five in each group) were treated with GII.4 type HuNoVs 2010, Sydney strains or an equal dose of PBS (control). Physical signs and diarrhea were observed for 7 days after HuNoVs challenge. HuNoVs RNA was detected by RT-PCR using stool samples collected every 24 h, and then the tree shrews were sacrificed to collect the small intestinal tissues. HE staining was used to observe pathological changes. The presence of HuNoVs antigen was tested by immunohistochemistry (IHC), and the saliva from tree shrews was analyzed for histoblood group antigen(HBGAs)by ELISA. Results No obvious infection symptoms or diarrhea were observed in the tree shrews after HuNoVs challenge. No HuNoVs RNA was detected in stool specimens, and no HuNoVs antigen was detected in the small intestine. No pathological changes were observed. HBGAs antigen was negative in the saliva of tree shrews. Conclusions Tree shrews are not susceptible to HuNoVs possibly because of the lack of HBGAs expression and common breeding conditions. The small animal model of tree shrews infected with HuNoVs remains to be further explored.

Abstract:Objective To investigate the effects of morroniside on angiogenesis-related factors and neurological function in rats 24 h after focal cerebral ischemia-reperfusion. Methods Healthy adult male SD rats were induced to develop occlusion of the middle cerebral artery by suture embolus. Then, the rats were randomly divided into a sham group,a model group, a morroniside-low group, a morroniside-intermediate group, and a morroniside low,moderate,and high-dose groups (30, 90, and 270 mg/ kg, respectively). Morroniside was administered 24 h after surgery, once a day for 7 days.By determining the scores of neurological behavior, the cerebral infarction volume ratios, and the expression of cortical angiogenesis-related gene proteins, were investigated to explore the effects of morroniside on angiogenesis-related proteins and neurological function in rats at 24 h after focal cerebral ischemia-reperfusion. Results Compared with those in the sham group, the scores of neurological behavior and the cerebral infarction volume ratios in the model group were significantly increased ( P <0. 001). After 7 days of morroniside administration, compared with that in the model group, the morroniside-high dose group exhibited a dramatic improvement of the neurological behavior scores ( P < 0. 01) and a significant reduction in the volume of cerebral infarction ( P <0. 05). Moreover, the expression of cortical angiogenesisrelated proteins CD34 and Ang-1 were significantly increased ( P <0. 05, P <0. 05) compared with those in the sham group.Compared with the model group, the expression of cortical angiogenesis-related proteins CD34 and Ang-1 in the group with high-dose morroniside was significantly increased ( P <0. 05, P <0. 01), and moderate-dose morroniside also promoted the expression of Ang-1 ( P <0. 05). Conclusions The time window for morroniside administration extends to 24 h after focal cerebral ischemia-reperfusion. The morroniside administration at a high dose can reduce the neurological behavior scores and the cerebral infarction volume ratio, while increasing the expression of the angiogenesis-related proteins CD34 and Ang-1.

Abstract:Objective To observe the protective effect of salvianolic acid A (SAA) on nephropathy and the expression of lipoprotein-associated phospholipase A2 (Lp-PLA2) in Zucker diabetes fatty rats ( ZDF). Methods According to the body weight and blood glucose, 40 male ZDF rats aged 11-12 weeks were randomly divided into five groups (n = 8 per group), namely, a model group, 0. 25, 0. 5, and 1 mg/ kg SAA groups, and a positive (Lp-PLA2 inhibitor at 25 mg/ kg) group. Meanwhile, another eight ZL rats of the same age were used as a normal group. Each drug administration group was given the corresponding drug once a day. Simultaneously, the model group and the normal group were administered normal saline of the same volume daily. After 12 weeks of continuous intervention, the renal microcirculation blood flow of ZDF rats was measured. Lp-PLA2 activity, blood urea nitrogen (BUN), and creatinine (Crea) in serum as well as transferrin (TRF), microalbumin (mALB), and retinol binding protein (RBP) in urine were analyzed. Lp-PLA2 mRNA and protein expression in renal tissue was also examined. Moreover, renal histopathological observation was performed. Results Compared with the control group, the pathological structure of the kidney was clearly changed and renal microcirculation blood flow was significantly decreased in the model group ( P <0. 01), whereas serum Lp-PLA2 activity and BUN levels and urinary TRF, mALB, and RBP were all notably increased ( P <0. 01). Moreover, the expression of Lp-PLA2 mRNA and protein in renal tissues was markedly increased ( P < 0. 01). The changes of these indicators were dramatically ameliorated after the administration of SAA and Lp-PLA2 inhibitor, especially in the groups with SAA at doses of 0. 5 and 1 mg/ kg. Conclusions SAA can improve the symptoms of DN in type 2 diabetic rats, which may be related to the inhibition of Lp-PLA2 activity and mass.

Abstract:Objective To explore an effective method for establishing a dual-target mouse model of angiogenesis in myocardial ischemic and atherosclerotic plaque. Methods Ten C57BL/6 J mice were fed basic forage as a control group, while ten ApoE^{-/ -} mice were fed high-fat forage as a model group. After 8 weeks, the model group was subcutaneously injected with isoproterenol at 100 mg/ kg once a day for 2 days, while the control group was subcutaneously injected with the same amount of physiological saline. Another 4 weeks later, the serum levels of TC, TG, LDL-C, and HDL-C were measured. HE staining was used to estimate the pathomorphological changes in aortic tissues and different regions of myocardium. Immunohistochemical staining was used to measure the density of new vessels in aortic plaques and different regions of the myocardium that were marked by CD31. Results The levels of serum TC and LDL-C in the model group were significantly higher than those in the control group ( P < 0. 05), while HDL-C was significantly lower than that in the control group ( P < 0. 05). HE staining revealed pathological changes typical of AS in the aorta of the model group.After 4 weeks of the subcutaneous injection of isoproterenol, pathological changes typical of myocardial infarction were observed in HE staining of myocardial tissue in the model group. CD31 immunohistochemical staining showed that the density of new vessels in aorta of the model group was significantly higher than that in the control group ( P < 0. 05). The expression of neovascularization in the myocardial ischemia zone was the highest in the model group, which was significantly higher than that in the myocardial infarction zone and normal myocardium zone ( P < 0. 05). Conclusions ApoE^{-/ -} mice with myocardial infarction induced by subcutaneous injection of isoproterenol under a diet of high-fat forage can be successfully used to establish a dual-target mouse model of angiogenesis in myocardial ischemia and atherosclerotic plaque.

Abstract:Objective To investigate the protective effects and mechanisms of action of rhodanine against cerebral ischemia-reperfusion injury. Methods A rat model of middle cerebral artery occlusion (MCAO) was created by exposure to ischemia for 2 h and reperfusion for 24 h. Neuroprotective score and cerebral infarction volume were used to evaluate the protective effects of rhodanine against cerebral ischemia-reperfusion injury. The superoxide dismutase (SOD) activity,the activity of glutathione peroxidase (GSH-Px) and the level of malondialdehyde (MDA) were detected. The expression of Bax, Bcl-2, and caspase-3 proteins were explored to reveal the brain-protective mechanism. Results Compared with those in the model group, the neurobehavioral scores and cerebral infarction volume ratios of the groups with low, medium, and high dose rhodanine were significantly lower ( P <0. 05). Compared with those in the model group, the activity levels of SOD and glutathione peroxidase (GSH-Px) in brain tissue were significantly increased in the groups with low, medium,and high dose rhodanine, while their malondialdehyde (MDA) levels were significantly decreased ( P < 0. 05). The expression levels of Bax and caspase-3 were decreased and the expression of Bcl-2 protein was increased in the low-,medium-, and high-dose groups. Conclusions Rhodanine has protective effects against cerebral ischemia-reperfusion injury, and its mechanism may be related to the improvement of SOD and GSH-Px activity, the decrease of MDA level, and an antioxidative effect. Upregulation of Bcl-2 protein expression and downregulation of Bax and caspase-3 protein expression may also be involved, indicating a relationship with neuronal apoptosis.

Abstract:Objective To investigate the effects and mechanisms of action of metformin combined with cisplatin on the apoptosis in human colon cancer HCT-8 cells. Methods Human colon cancer HCT-8 cells were divided into four groups: metformin group (MET), cisplatin group (DDP), metformin combined with cisplatin group (MET+DDP),and blank control group (CK). The proliferative ability of HCT-8 cells was determined by an MTT experiment at 24,48 and 72h. The apoptosis rate was determined by flow cytometry at 48 h. Western blotting was used to reveal the expression of apoptosis-related proteins such as bcl-2, Bax and caspase-3 (activated),as well as proteins of the AKT/ GSK-3β pathway at 48 h. Results The cell proliferative ability in the MET+DDP group was weaker than that of the other three groups ( P <0. 05). Moreover, its apoptosis rate was higher than those of the other three groups ( P <0. 05). The expressions of Bax and caspase-3 (activated) was higher than that of the other groups, the expression of Bcl-2, p-AKT and p-GSK-3β was lower than that of the other groups, and the expression of AKT and Gsk-3 was relatively consistent among the four groups.Conclusions Metformin and cisplatin in combination can reduce the activity of the AKT/ GSK-3β signaling pathway, and change the expression of Bcl-2 family proteins,to promote the apoptosis of human colon cancer HCT-8 cells. They also promote the p-caspase-3 conversion to caspase-3 p-caspase-3 conversion to caspase-3 (activated), metformin and cisplatin can thus play a role in promoting the apoptosis of HCT-8 human colon cancer cells.

Abstract:Objective This experiment was conducted to study the effect of safflower injection on the expression of ER stress-related molecules, GRP78, CHOP, and high-sensitivity C-reactive protein in the brain of rats with middle cerebral artery occlusion(MCAO),to explore the effect of safflower injection on the treatment of stroke by modulating inflammation through ERS, so as to provide an experimental basis for the clinical application of safflower injection in the treatment of acute cerebral infarction. Methods A rat MCAO model was established through a modified suture method,followed by intravenous injection of safflower. Rat brain tissues were examined by immunohistochemistry to determine the expression of ERS-related markers CHOP and GRP78. RT-qPCR was performed to determine the expression of GRP78 mRNA and CHOP mRNA. ELISA was used to determine the expression level of high-sensitive C-reactive protein. Results Twelve hours after stroke, the cerebral infarction area ratio of the safflower injection group was significantly lower than that of the model group. The neurological deficit scores of the safflower injection group were significantly lower than those of the model group. The expression of GRP78-positive cells in the safflower injection group was significantly higher than that of the model group, while the expression of CHOP in the safflower injection group was significantly lower than that of the model group. Compared with that in the model group, there was no significant difference in the expression of GRP78 mRNA and CHOP mRNA in the safflower injection group. The CRP content of the safflower injection group was significantly lower than that of the model group. Conclusions Safflower injection has protective effects against stroke, and the mechanism of ERSrelated to CRP may be mediated by upregulation of GRP78 and downregulation of CHOP.

Abstract:Objective To explore a method to construct a rat model with qi stagnation and blood stasis chronic salpingitis, to study its feasibility and establish a method for its primary evaluation, and to lay the foundations for future experiments. Methods SD rats were subjected to predictable repeated stress stimulation (e.g., sound, light, electricity,and clip tail stimulation) and bacteria were injected into the oviduct to construct the model with qi stagnation and blood stasis chronic salpingitis. Symptoms, and observation, pathological, and blood rheology indexes were used to primarily evaluate the rat model. Results Compared with the control group, the combined qi stagnation and blood stasis with chronic salpingitis model group’s total mark rose significantly Qi( P <0. 01), 80% of the oviduct blood vessels exhibited swelling and distension, and 60% of their walls showed thickening. Pathological analysis revealed the infiltration of inflammatory cells in the oviduct lumens of the rat model with qi stagnation and blood stasis chronic salpingitis, and epithelial mucosal ablation was accompanied by substantial inflammatory cell infiltration surrounding the oviduct. Blood rheology analyses demonstrated very significantly increased plasma viscosity, whole blood viscosity, reduction viscosity, and erythrocyte rigidity index and aggregation index in the combined rat model group ( P < 0. 01). Conclusions Our comprehensive analysis reveals that the rat model with qi stagnation and blood stasis chronic salpingitis is roughly consistent with the dynamic characteristics of blood stasis and the pathological status of chronic inflammation, being similar to the human clinical manifestations. The method for constructing this model is feasible.

Abstract:Objective To observe the changes of matrix metalloproteinases (MMPs), TIMP2, and inflammatory factors TNF-α and IL-1β in the myocardium of rats with acute myocardial infarction (AMI) treated with rosuvastatin.Methods Healthy SD rats were selected to establish an AMI model by ligation of the left anterior descending coronary artery.They were divided into four groups: a control group (isolation of anterior descending branch without ligation, n = 10); an acute myocardial infarction group (AMI modeling group); a rosuvastatin group (n =13, AMI treated with rosuvastatin at 10mg/ (kg·d) after modeling); and a losartan group (n = 11, AMI treated with losartan potassium at 5 mg/ (kg·d) after modeling). Immunohistochemical staining and RT-qPCR were used to determine the expression levels of MMP2, MMP9, and inhibitor TIMP2 in rat cardiac tissues after AMI modeling. Western blotting was used to determine the changes of MMP2,MMP9, TNF-α, and IL-1β in myocardium. Results The results of immunohistochemical and RT-qPCR analyses showed that, compared with those in the AMI group, the MMP2 and MMP9 protein/ mRNA expression levels in rat myocardium in the rosuvastatin and losartan groups were decreased ( P < 0. 05). Regarding the western blotting results, compared with that in the control group, the expressions of MMP9, MMP2, TNF-α, and IL-1β proteins in the AMI group were significantly increased( P <0. 05). Moreover, compared with those in the AMI group, the expression levels of MMP2, MMP9, TNF-α, and IL-1β in the myocardium of the rosuvastatin and losartan groups were decreased ( P <0. 05). Conclusions Myocardial fibrosis factors and inflammatory factors are increased after AMI modeling in rats. Rosuvastatin can improve the cardiac function to some extent by inhibiting myocardial fibrosis and reducing the expression of inflammatory factors.

Abstract:Objective To investigate the effects of exposure of the head of rats to different doses of ionizing radiation, to establish an appropriate animal model of cognitive function damage caused by such radiation, and to provide a research basis for studying the mechanism of cognitive function damage due to ionizing radiation and the development of radiation-protective agents. Methods Twenty male SPF rats were randomly and equally divided into a control group (C group), a 10 Gy irradiation group (10 Gy IR group), a 20 Gy irradiation group (20 Gy IR group), and a 30 Gy irradiation group (30 Gy IR group). The group C was left untreated, while the other three groups were irradiated with 10,20, and 30 Gy electron beams, focused on the heads of the SD rats. The mortality of the animals was calculated using survival rates at 30 days after the irradiation and spatial memory was assessed using the Morris water maze. The concentrations of glutathione (GSH) and malondialdehyde (MDA) in rat brain cortical homogenate were determined by chemical colorimetry. An enzyme-linked immunosorbent assay (ELISA) was used to detect 8-hydroxydeoxyguanosine (8-OHdG). Brain pathological sections were analyzed by HE staining. Results The rats in the 30 Gy IR group exhibited irregular back hair, excessive salivation, and other symptoms, along with severe hair loss on the head and neck. At 30 days after the irradiation, no animal died in group C. The mortality rates of both the 10 and 20 Gy IR groups were 20%, while that of the 30 Gy IR group was 40%. The results regarding positioning and navigation in the Morris water maze experiment showed that the latency of the 30 Gy IR group in the 1-5-day training period was significantly higher than in the other three groups ( P <0. 05). The spatial search experiment showed that the residence time in the original platform quadrants of the 30 Gy IR group was significantly shorter than in the other groups ( P <0. 05); the number of platform crosses in this group was also significantly reduced ( P <0. 05). The swimming distances in the original platform for the 10, 20, and 30 Gy IR groups were significantly shorter than for the group C ( P < 0. 05). Moreover, the results of oxidative stress-related indicators showed that the concentrations of GSH, 8-OHdG, and MDA in the 30 Gy IR group were significantly different from those in group C ( P <0. 05). The difference of MDA concentration between the 20 Gy IR group and group C was statistically significant ( P <0. 05). Pathological examination also showed that the tissue damage of the 20 and 30 Gy IR groups was more severe than that of the 10 Gy IR group, and there were significant pathological changes such as cell apoptosis and tissue necrosis. Conclusions This experiment shows that a rat model of cognitive dysfunction can be created by a single 30 Gy electron beam irradiation to the head.

Abstract:Objective To explore the mechanism by which puerarin stimulates osteogenesis and bone formation through the ERK1/2 and p38 MAPK signaling pathways. Methods Adult osteoblasts (MC3T3-E1) cell culture was used in this study.The proliferative ability and the influence of growth curve of the cells after added different concentrations of puerarin were compared by MTT staining. The effect of puerarin on the differentiation of osteoblasts was assessed by measuring alkaline phosphatase activity and the effect of puerarin on bone formation was analyzed by measuring calcium deposition. Through the addition of ERK1/2 blocker PD8089 and p38 inhibitor SB203580, the mechanisms of involvement of the ERK1/2 and p38 MAPK signaling pathways in stimulating osteogenesis and bone formation were analyzed. Western blotting was used to detect the expression of bone morphogenetic protein-2 (BMP-2). Results Different concentrations of puerarin promoted the proliferation of osteoblasts to different degrees. The effect of puerarin at 0. 1 mol/ L was the most significant. The trend of proliferation was not pronounced on the first and third days compared with the level in the blank group, but significant differences emerged between the fifth and seventh days. Puerarin activated alkaline phosphatase activity and promoted the differentiation of primary osteoblasts. It also promoted calcium deposition and stimulated bone formation. After using the ERK1/2 blocker PD8089 or blocking the p38 MAPK signaling pathway with the inhibitor SB203580, cell proliferation, alkaline phosphatase content, and calcium deposition were lower than those of the puerarin group. The expression of BMP-2 in group puerarin (T) was higher than that of the control group ( P <0. 05), and that of group puerarin+PD 8089(T+PD)was lower than that of group T, while the amount of calcium deposition in the group puerarin+SB 203580(T+SB)was significantly lower than that in group T ( P <0. 05), and there was a decrease ( P <0. 05)in the BMP-2 expression compared with that in the control group. Conclusions In the cell cycle in bone, puerarin and the ERK1/2 and p38 MAPK signaling pathways play regulatory roles in bone differentiation and bone formation.

Abstract:Objective To establish animal models of spinal cord injury of different degrees of severity, namely,MASCIS and IH, and to study the relationship among the degree of damage, modeling mode, behavior, and anatomical results. Methods Eighty female SD rats were randomly and equally divided into four groups: NYU 12. 5 mm, NYU 25mm, IH 150 kdyn force, and IH 200 kdyn force models. Their rates of successful model establishment, BBB score, grid walking evaluation, Hargreaves test, and total cross-sectional area of the spinal cord at the injury epicenter were evaluated.Results The success rates in the two modeling methods were similar. The BBB scores of all animals were reduced to 0points at 0 to 3 days after spinal cord injury. As time passed, the BBB scores gradually recovered and reached a plateau at 4to 6 weeks. The NYU 25 mm group scored the lowest, the IH 200 group had an intermediate score, and the IH150 and NYU 12. 5 mm groups scored similarly and highest. The grid walking score increased with increasing injury severity. The Hargreaves test results were more affected by motor function. Dyskinesia was inversely proportional to the residual amount of tissue at the center of the injury. Conclusions Both MASCIS and IH models are appropriate for research on the pathophysiological mechanisms of spinal cord injury, but appropriate modeling parameters should be selected according to the purpose of the experiment.

Abstract:Objective To investigate the effects of wogobin on the proliferation, migration, and invasion of gastric cancer cells and their molecular mechanisms. Methods Different concentrations (20, 50, and 100 mol/ L) of wogobin were used to treat the hepatoma cell line SGC7901 cells for 24, 48, and 72 h. In the blank control, no treatment was performed. MTT assay was used to determine the cell proliferation. The migration ability of cells was observed by a scratch test. The Transwell method was used to observe cell invasiveness. Cell cycle and apoptosis were determined by flow cytometry. The mRNA and protein expression levels of cellular MMP2, MMP9, ICAM-1, and TIMP2 were determined by RT-qPCR and western blotting, respectively. Results MTT assay showed that the different concentrations of wogobin inhibited the hepatoma cell line SGC7901 in a concentration- and time-dependent manner. After 48 h, wogobin arrested the cell cycle at S-phase and induced the apoptosis in gastric cancer cells ( P <0. 05). Transwell assay showed that wogobin inhibited the invasion of the SGC7901 cells in a concentration-dependent manner. The RT-qPCR and western blotting analyses showed that wogobin inhibited the mRNA and protein expression of MMP9, MMP2, and ICAM-1 in SGC7901cells, but upregulated TIMP2 ( P <0. 05). Conclusions Wogobin inhibites the proliferation, migration, and invasion of the hepatoma cell line SGC7901 cells and induces its apoptosis.

Abstract:Objective To investigate the vaginal irritation and skin sensitization associated with metronidazole vaginal gel, and to provide evidence for its clinical application. Methods An irritation test was carried out on rabbits for vaginal administration. After 7 days of continuous administration, possible vaginal irritation was observed and recorded. A skin allergy test was administered on guinea pig skin on days 0, 7, and 14. Sensitization was initiated on day 28 to observe guinea pig skin or systemic allergic reactions. Results The topical administration of metronidazole vaginal gel did not cause vaginal irritation or allergic reactions. Conclusions Metronidazole vaginal gel is safe for the treatment of bacterial vaginosis.

Abstract:Objective To provide a reference for improving the quality of laboratory animals in Sichuan Province,we reviewed and summarized the results of microorganism and parasite sampling in laboratory animals in the Sichuan area from 2011 to 2015. Methods We sampled and tested the laboratory animal production units in Sichuan Province, and reported on these using current national laboratory animal guidelines. In addition, we analyzed the quality of laboratory animals of different microbiological grades in recent years within Sichuan Province, including mice,rats, guinea pigs,rabbits, dogs, monkeys, and piglets. Results the sample sizes and inspectial frenqency annually of animal samples in these 4 years were basically stable. In 2012, helminths were detected in SPF rats in two units, with a positive rate of 22. 5%. The positive rates of Staphylococcus aureus were 5. 0%, 5. 0%, 40. 0%, and 11. 4% in different units for 4 consecutive years. In 2015, parvovirus RV and H-1 in rats were positive in three units, with positive rates of 34. 3% and 28. 6%, respectively. In 2012 and 2013, ectoparasites were detected in different units of SPF mice, with positive rates of 7. 1% and 12. 9%, respectively. In the helminthes inspection,the positive rate of the samples was 10% in 2012,1. 4% in 2013 and 7. 5%in 2015.In 2011, Klebsiella pneumonia was positive in one unit, with a positive rate of 1. 4%. The guinea pigs samples fet the standards totally in recent years. In 2013, Toxoplasma gondii was positive in rabbits in one unit, with a positive rate of 10%. In 2012, the positive rates of pathogenic dermal fungi and Toxoplasma gondii in dogs from two units were 5. 0% and 2. 5%, respectively. In 2011, the monkeys in two units tested positive for pathogenic dermal fungi, with a rate of 9. 5%. In 2011 and 2012, the same single unit tested positive for Shigella spp. for two consecutive years, with rates of 2. 4% and 2. 5%, respectively. The cercopithecine herpesvirus type 1 was found in one unit in 2012, with a positive rate of 2. 5%. In 2015, in the first examination of piglets, only Brucella and Toxoplasma were tested, with a negative result.Conclusions Quality monitoring and sampling inspection of laboratory animals are important to guarantee the quality of laboratory animals. They also play a vital role in improving the quality of laboratory animals in Sichuan Province, reducing the differences between Sichuan and developed area, and promoting the development of the Sichuan laboratory animal industry.

Abstract:Neurodegenerative diseases are common conditions globally that involve the degeneration of neurons, for which no effective curative measures are available. Stem cells have the potential to undergo self-renewal and directional differentiation, potentially providing new therapeutic method for the recovery of brain function in neurodegenerative diseases. In recent years, with the development of stem cell biology, it has been found that adipose stem cells are pluripotent stem cells that develop from the middle germ layer, which can differentiate into different lineages upon exposure to specific growth factors and environmental conditions.They have the benefits of being easily accessible to researchers and simple to culture. Therefore, adipose tissue-derived stem cells have unique advantages in treating neurodegenerative diseases. This article provides an overview of neurodegenerative diseases, the biological characteristics of adipose-derived stem cells, and the conditions for differentiating them into the components of neural systems.

Abstract:The long noncoding RNA HOXA11 antisense RNA (HOXA11-AS) was recently discovered in a mouse embryo cDNA library using the probe from the sense HOXA11 cDNA sequence. Since then, it has been found to play important roles in human cervical cancer, gastric cancer, and gliomas. HOXA11-AS can promote tumor malignancy (i.e.,proliferation and metastasis) by interacting with various miRNAs and proteins such as EZH2, providing new potential routes for the potential treatment of cancers.

Abstract:In recent years, CRISPR/ Cas9 technology has become the hottest gene editing technology through its advantages of being flexible, efficient, cheap, and easy to operate, and its ability to edit multiple sites at the same time.The unique CRISPR/ Cas9 RNA-guided mechanism of targeted editing of DNA has not only expanded our understanding of the genetic regulation of organisms, but also demonstrated superior convenience. In the last ten years, CRISPR/ Cas9 has been widely used in various biological and medical research fields and has developed rapidly. Especially in the field of medicine, it has shown great potential. This paper summarizes recent advances in CRISPR/ Cas9 technology, and the mechanism of action and application in medicine of the CRISPR/ Cas9 gene editing system, to provide a reference for the application and optimization of this technology.

Abstract:The lysosome, an organelle rich in acidic hydrolase, is the main degradation organelle in the cell and is involved in cell processes such as secretion, plasma membrane repair, cell signaling, and energy metabolism. Lysosomal dysfunction may be related to the development of various diseases. In recent years, studies have shown that lysosomal dysfunction may be involved in the development of chronic kidney disease(CKD): lysosomal dysfunction of podocytes interrupts autophagy, inducing transdifferentiation and damage of podocytes. Lysosomal damage mediates apoptosis in proximal tubule epithelial cells via oxidative stress. Lysosomal damage leads to reduced degradation of type I collagen in mesangial cells. Renal endothelial cell dysfunction is closely related to the abnormal expression of lysosomal cathepsin-S and α-galactosidase-A. In summary, lysosomal dysfunction is closely linked to the pathology of intrinsic renal cells. Elucidation of the function of the lysosome in intrinsic renal cells should provide new ideas for clarifying the mechanism of intrinsic renal cell damage in chronic kidney disease.

Abstract:The blood-brain barrier (BBB) is a dynamic barrier between the peripheral circulatory system and brain tissue, whose permeability to polypeptide drugs has a direct impact on the effect of drugs on the central nervous system (CNS). With the development of neuroscience and genetic engineering technology, a large number of protein and polypeptide drugs have been applied to the treatment of nervous system diseases. Many protein and polypeptide drugs have biological activities on the CNS, but the BBB prevents these substances from entering the CNS and exerting therapeutic effects. The mechanisms of transport of peptides and ways of promoting the penetration of the BBB by polypeptide drugs are summarized in this article and proteins or peptides known to penetrate the BBB are introduced. It is hoped that this paper will provide some direction for the development of CNS drugs.