Objective To explore the regulatory mechanism of emodin on pain behavior in a mouse model of osteoarthritis based on mitochondrial key genes.Methods Thirty C57BL/6J mice were randomly divided into the control group,the osteoarthritis(OA)model group and the emodin-treated(OA+emodin)group,10 mice per each group.The mice in the OA group and the OA+emodin group were intra-articular injection of complete Freund's adjuvant(20 μL)in knee to establish the OA model,and mice in the OA+emodin group were treated by intraperitoneal emodin(10 mg/kg)injection.After behavioral testing,knee tissue of mice was collected for hematoxylin-eosin staining.Western blot analysis was used to detect expression levels of proinflammatory factors interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α)and mitochondria-related proteins NADH dehydrogenase(ubiquinone)flavoprotein 1(NDUFV1),cytochrome C oxidase subunit 5B(COX5B),cytochrome C oxidase assembly protein COX15 homolog(COX15),NADH dehydrogenase(ubiquinone)1 alpha subcomplex subunit 10(NDUFA10)in knee tissue.Results Compared with the control group,mice in the OA group showed decreased mechanical nociceptive threshold(PWT),reduced latency and distance in rotarod test(P＜0.05).Compared with the OA group,mice in the OA+emodin group showed increased PWT,latency,and distance(P＜0.05).In the control group,the structures of cartilage and subchondral bone were intact,while in the OA group,the cartilage was thinner and the subchondral trabeculae was deteriorated.The treatment with emodin alleviated cartilage degeneration.The expression levels of IL-1β,TNF-α,COX15 and NDUFA10 were increased while expression levels of NDUFV1 and COX5B were decreased in the OA group compared with the control group.The emodin treatment restored the above-mentioned protein expression levels(P＜0.05).Conclusion Emodin can alleviate pain behavior in OA mice by regulating the expressions of inflammatory factors and mitochondrial related proteins.

Objective:To investigate the role and mechanism of chemokine receptor type 4 (CXCR4) in renal injury and fibrosis caused by calcium oxalate crystals in mice.Methods:In June 2021, Fifteen male C57/BL6 mice were divided into control group (5 mice), model group (5 mice), and AMD3100 intervention group (5 mice) by random number table method. In model group and AMD3100 intervention group, glyoxylate (100 mg/kg) was injected intraperitoneal for 7 consecutive days for modeling. Meanwhile, the AMD3100 intervention group was also given intraperitoneal injection of AMD3100 (1 mg/kg) for 7 days. The control group was continuously injected with equal volume saline intraperitoneally. After 7 days, peripheral blood was collected from each group to determine the levels of serum urea nitrogen (BUN) and creatinine (Scr) to assess the renal function; HE, Von-Kossa, Picrosirius Red staining was also taken from the left kidney to observe the pathological changes of renal tissue. CXCR4, transforming growth factor β1 (TGF-β1) were detected by immunohistochemistry and western blot. The expression levels of PI3K/AKT pathway-related proteins were detected by western blot.Results:The results of biochemical indexes showed that the serum Scr [(108.03±13.56) μmol/L vs. (39.50±4.48)μmol/L, P<0.01)] and BUN[(5.66±0.48)mmol/L vs. (0.77±0.10)mmol/L, P<0.01) levels were significantly increased in model group compared with the control group. The AMD3100 intervention group was significantly lower than the model group in terms of Scr [(65.77±3.27)μmol/L vs. (108.03±13.56)μmol/L, P<0.05) and BUN [(2.97±0.44)mmol/L vs. (5.66±0.48)mmol/L, P<0.05) levels. The results of kidney pathology in mice showed that renal tubules were significantly dilated with inflammatory cell infiltration in the model group compared with the control group, and a large number of calcium oxalate crystals and collagen fibers were deposited. The extent of kidney damage, calcium oxalate crystals and collagen fibers deposition were significantly reduced in the AMD3100 intervention group compared with the model group. The results of western blotting showed that the relative expression of CXCR4(0.639±0.019 vs. 0.158±0.012, P<0.01) and TGF-β1(0.698+ 0.018 vs. 0.314+ 0.015, P<0.05) was significantly increased in the model group compared with the control group. The relative expression of CXCR4(0.322±0.231) in the AMD3100 intervention group compared with the model group (0.322±0.231 vs. 0.639±0.019, P<0.05) and TGF-β1(0.445+ 0.017 vs. 0.698+ 0.018, P<0.05) were significantly decreased. The results of immunohistochemical staining showed the trend of CXCR4 and TGF-β1 expression in each group consistent with the results of protein blotting assay. Western blotting results showed that the expression of p-PI3K (0.613±0.016 vs. 0.213±0.011, P<0.01) and p-AKT(0.149±0.013 vs. 0.047±0.014, P<0.01) was significantly increased in the model group compared with the control group. The expression of p-PI3K in the AMD3100 intervention group compared with the model group (0.292±0.020 vs. 0.613±0.016, P<0.05) and p-AKT (0.098±0.021 vs. 0.149±0.013, P<0.05)was significantly decreased. Conclusion:CXCR4 inhibits calcium oxalate crystal-induced kidney injury and interstitial fibrosis in mice by targeting the PI3K/AKT pathway.

GLKs (GOLDEN 2-LIKEs) are a group of plant-specific transcription factors regulating the chloroplast biogenesis, differentiation and function maintains by triggering the expression of the photosynthesis-associated nuclear genes (PhANGs). The GLKs also play important roles in nutrient's accumulation in fruits, leaf senescence, immunity and abiotic stress response. The expression of GLK genes were affected by multiple hormones or environmental factors. Therefore, GLKs were considered as the key nodes of regulatory network in plant cells, and potential candidates to improve the photosynthetic capacity of crops. Since numerous researches of GLKs have been reported in plants, the biological function, molecular mechanism of GLKs genes and its applications in breeding were summarized and a GLK-mediated signaling network model was developed. This review may facilitate future research and application of GLKs.
Chloroplasts/genetics* ; Gene Expression Regulation, Plant ; Photosynthesis/genetics* ; Plant Breeding ; Transcription Factors/metabolism*

Objective:To compare the injury of renal blood vessels using different puncture pathways and access sizes.Methods:Between April 2018 and June 2019, eighty fresh pig kidneys were selected to perform percutaneous puncture and dilation, which was used to compare the injury of renal blood vessels with different puncture pathways and access sizes. The puncture pathway included the centerline of the normal renal pyramid (A), centreline of one side pyramid of the fused renal pyramid (FRP) (B), midline of the entire FRP (C) and midline of the renal column (D). The access size included F8, F12, F16, F20, F24 and F30. Histopathological methods were used to analyze the injury of renal blood vessels.Results:The puncture through paths A and B mainly caused injury to the grade Ⅴ and Ⅵ arteries in renal cortex. The puncture often directly injures the grade Ⅳ artery in path C. The puncture often simultaneously injures the grade Ⅲ-Ⅵ arteries in path D. Grade Ⅲ artery injury began to occur when paths A, B, C, and D were dilated to F30, F24, F16, and F12, respectively. The degree of arterial injury among the four different puncture pathways was significantly different in F8, F12, F16, F20, F24 and F30 ( P<0.05). Statistical differences were found between paths A and D in F12, F16, F20, F24 and F30 ( P<0.05), and between paths A and C in F16, F20 and F24 ( P<0.05). No significant difference was found between paths A and B in all access sizes ( P>0.05). Compared with F8, the degree of arterial injury of the F30 in path A and the F24 and F30 in path B were increased significantly ( P<0.05). Conclusions:Vascular injury in path D was the most serious followed by that in path C. Relatively little vascular injury can be achieved in paths A and B. The vascular injury increased when the path B was dilated to F24, while the path A needed to be dilated to F30.

Objective To evaluate the safety of fibrin sealant (FS) intraperitoneal injection in SD rats .Methods 80 male and female SD rats were randomly divided into four groups (0 ,85.5 ,171 .0 ,342 .0 mg/kg) by body weight .All rats were in-traperitoneally injected with vehicle or FS daily for 14 days followed by a 28-day recovery period .The clinical signs ,hematolog-ical and biochemical indices were measured .The pathology were observed .Results Increase of white blood cell count (WBC) and decrease of fibrinogen (FIB) in d 14 were found in 171 .0 mg/kg and 342 .0 mg/kg dosage groups .Furthermore ,the tend-ency of weight increase of spleen were found in 171 .0 mg/kg and 342 .0 mg/kg dosage groups .Pathological exams of peritoneal cavity found that there were granulation tissues containing FS in some of the rats in 342 .0 mg/kg group .All of these changes got reversed after the recovery period .Conclusion The safety dose in this study is considered to be 85.5 mg/kg ,and the toxic-ity dose is 171 .0 mg/kg .The target toxicity systems or site of FS in SD rats are hematological system ,immune system and in-jection site .The toxic effects of FS are reversible .

The quality management of drug research,development,registration,production and marketing strengthened by good practice for pharmaceuticals ensure the drag safety,effectiveness and quality control.Teaching of new drug research and evaluation in compliance with good practice for pharmaceuticals will be of value in making teaching content close to actual work,extending the students'knowledge and training student's good habits in scientific study.

BACKGROUND: Besides being a basic growth factor crucial to maintain and promote the development, differentiation and survival of the central nervous system, nerve growth factor(NGF) also plays an important role in the repair of injured peripheral nerves.OBJECTIVE: To investigate the effect of the muscular injection of NGF on the regeneration and functional recovery of rat sciatic nerve after crush injury.DESIGN: A randomized controlled pilot study in rats with repeated observation and measurement.SETTING: Center for new drug evaluation in a military medical university.MATERIALS: This study was performed in the Center for New Drug Evaluation, Department of Basic Medicine, Second Military Medical University during the period from July 1999 to March 2000, using 40 SD rats weighing 200 to 250 g(of either sex of half number) provided by the Sino-British SIPPR/BK Lab Aninal Ltd (Shanghai).METHODS: Forty rats were randomized into high-, mid- and low-dose NGF treatment groups, normal control group and model control group. The sciatic nerves were clamped at 6 nm distal to the sciatic notch to induce a 4-mm-wide area of crush injury. In the high-, mid-; and low-dose NGF groups, the rats were given NGF at 8, 4 and 2 μg/kg per day(corresponding to 1.6 × 10 3, 8 × 10 2 and 4 × 10 2 IU/kg per day) respectively via the muscular injection for 56 consecutive days.(NCVs) and sciatic function index(SFI) at different time points after the RESULTS: Compared with that of the model control group, the NCVs significantly increased in the high-dose NGF group 35 and 56 days after the injury,and in the mid-dose NGFgroup at 35 days(t=2.32-5.14, P ＜0.05-0.01 ). The SFIs significantly increased in all NGF-treated groups at 14 days ( t = 2. 29-6.28, P ＜ 0.05-0.01 ), with the recovery most conspicuous in high-dose NGF group; No significant difference in the SFIs was found between the NGF-treated groups on the 56th day. Morphological examination of the tissues identified no significant difference in the nerve myelin sheaths and axons in NGF-treated groups as compared with the normal control group,while in the model control group, myelin sheath dislocation with unclear microstructure was observed, accompanied by Schwann cell degeneration and necrosis.CONCLUSION: NGF promotes the repair of the damaged nerve myelin sheath and axon and stimulates nerve fiber regeneration and function recovery of the crushed rat sciatic nerves.

Objective:To investigate the long-term toxicity of recombinant human interleukin-11(rhIL-11) in cynomolgus. Methods: Eighteen cynomolgus were randomized into 4 groups: control group(2/sex), low dose group(2/sex), medium dose group(2/sex)， and high dose group(3/sex). The drug groups were sc adminstered 0.1, 0.3 and 1.0 mg/kg of rhIL-11 for 90 days with a 30-day recovery period. The clinical signs were observed, electrocardiogram, hematological, biochemical, urinary and immunological parameters were measured, organ masses were weighed, bone marrow and pathological histology were observed. Results: The food consumption, body mass of the drug groups were decreased, the body temperature was increased transiently. One of the low dose group showed restricted movements and tremors. One of the high dose group vomited and another died. Reduced red blood cell(RBC) count, hemoglobin(Hb) concentration, hematocrit(Hct), mean corpuscular volume(MCV), mean corpuscular hemoglobin(MCH), and mean corpuscular hemoglobin concentration(MCHC), dose-related increase of platelet(Plat) counts were present in drug groups. Biochemical examinations revealed dose-related decreases in serum alanine aminotransferase(ALT), aspartate aminotransferase(AST), lactate dehydrogenase(LDH), total proteins(TP) and albumin(Alb) increases in serum alkaline phosphatase(ALP) levels. Positive antibody responses were seen and circulatory immune complex(CIC) was significantly increased in all drug groups. Hypertropy of marrow megakaryocyocytes was noted in the medium and high dose groups. The heart and liver masses were slightly increased in all treatment groups. Treatment-related microscopic findings included dose-related degeneration in the liver and the kidney. The adverse effects were reversed by the end of the recovery period. Conclusion: The target organs and systems are blood, liver, kidney, immmue system and bone marrow. The toxicity injuries were reversible and the no-toxic-effect level is 0.1 mg/kg.

Objective　To research the direct electrophysiological evidence of discomplete spinal cord injury (SCI) and the effect of 4-aminopyridine on it. Methods　Motor evoked potentials (MEPs), both spinal cord recorded MEPs (scMEPs) and extracellularly recorded MEPs (exMEPs) were recorded and characterized on a T13 epidural electrode (scMEPs) and an extracellular microelectrode (exMEPs) for 10 normal rats and 40 rats with lesions of various severity (sham, 35?g*cm force (gcf), 70?gcf, 100?gcf impact injury) at the T8-T9 cord using the Allen's drop model. The incline plane and Tarlov techniques were used to assess clinical neurological function. Results　MEPs in the normal rats were elicited by applying transcortical suprathreshold stimulation consisting of 3-4 early negative peaks (N1, N2, N3 and N4) followed by several late waves. The N1 and N2 peaks were largest in the anterior and ventrolateral funiculus, respectively, which was indicative of extrapyramidal pathways. The 100?gcf impact injuries and the cord transection abolished the MEP distal to the lesion, whereas the 35?gcf injuries resulted in a latency shift and amplitude decrement of the MEP peaks. Eighteen of the 20 rats with 70?gcf injuries showed clinical paraplegia. Among them, 7 rats had neurophysiological evidence of residual conduction pathways through the lesioned cord segment, such as the presence of N1 and N2 peaks in the scMEPs or exMEPs. After 4-aminopyridine (4-AP) administrations (1?mg/kg), the amplitude of the spared exMEP increased significantly and spread more widely. Conclusions　MEPs evoked by transcortical stimulation travel mostly in the extrapyramidal tract. MEP monitoring could provide an excellent method of detecting the functional integrity of the motor tracts after SCI, and could even detect spared motor fibers after discomplete SCI. Furthermore, the use of 4-AP or other K+ channel blocking agents may be a potential treatment for patients with chronic moderate to severe SCI.

Shiwei Dangguiyin(SWDGY)is mainly composed of Radix Angelicae Sinensis,Radix Adenophorae,Radix Notogenseng,Radix Bupleuri,etc. Oral administration of SWDGY could significantly inhibit the metatarsal swell- ing eaused by dimethylbenzene in rats,raise the pain threshold in hot-plate test and depress the torsive reaction caused by acetic acid in mice.In vitro SWDGY exerted bacteriostatic and bacteriocidal effects on Staphylococcus aureus,Bacil- lus pyocyaneus,Escherichia coli,Streptococcus A,B and C.It was shown that SWDGY possessed anti-inflammatory,analgesic and antiseptic effects in vitro.In mice LD_(50) of SWDGY by oral administration was more than 840g/kg.Affer cral adminstration in a daily dose of 189.Sg/kg continuously for one month in rats, no toxic reactions appeared,This dosage was 118.6 times as much as the clinical one.