OBJECTIVE:To establish a method for the content determination of L-epicatechin in Actinidiae arguta. METHODS:HPLC was performed on the column of Kromasil C18 with mobile phase of acetonitrile-0.2%Acetic acid solution(15:85,V/V)at a flow rate of 1.0 ml/min,detection wavelength was 210 nm,column temperature was 25 ℃,and the volume injection was 10 μl. RESULTS:The linear range of L-epicatechin was 10.47-167.52 μg/ml(r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 2%;average recovery was 98.07%-101.71%(RSD=1.39%,n=6). CONCLUSIONS:The method is simple, accurate and reliable,and suitable for the content determination of L-epicatechin in A. arguta.

Objective To investigate the effects of signal transducer and activator of transcription 3 (STAT3) RNAi on the content of reactive oxygen species (ROS) and the DNA damage in glioma cells.Methods Glioma cells of the line U251 cells were cultured and transfected with STAT3 RNAi plasmid (pSilencer2.1-STAT3,STAT3 group) and pSilencer2.1-GFP (GFP control group) respectively.Part of the U251 cells were irradiated with γ-rays of 60Co as positive control group of smear phenomenon.The levels of ROS and malondialdehyde (MDA) in the cells were detected 24,48,and 72 h later by flow cytometry and fluorescence chamoluminescence analyzer,respectively.The DNA damage in the transfected U251 cells was examined by using single cell gel electrophoresis assay,and the cell cycle distribution was examined using FACS PI staining 12,24,and 36 h later.Results At 24 h after the transfection,the ROS level of the siSTAT3-transfected ceils was 8.91 times that of the control group (F = 89.296,P < 0.05),and returned to the normal level 48 h later.There were not significant differences in the MDA level of the cells 24,48,and 72 h later between the siSTAT3 group and siGFP group.Compared with the 8 Gy irradiation positive group with obvious smear phenomenon,smear phenomenon was shown in part of the ceils in the siSTAT3 group 6 h later,became less 12 h later,and disappeared completely 24 h later.Compared with the control group,lag of S stage rate was 17.22% and the lag of G2/M stage rate was 6.4% 12 h later in the siSTAT-transfected group,and the G0/G1 stage lag rate was 18.44% 24 h later,and the lag of S stage rate was 17.99% 36 h later.Conclusions Inhibition of STAT3 results in the change of oxidoreduction status in glioma cells,as well as damage and reparation of DNA.

Objective: To investigate the relationship between cervical curvature (CC) and spinal drift distance after laminectomy with lateral mass screw fixation and the influence on the operative effect. Methods: From October 2016 to December 2017, a total of 85 patients with cervical spondylotic myelopathy (CSM) underwent laminectomy with lateral mass screw fixation in handan central hospital, and 78 patients were followed up completely.After the operation, according to the Harrison method, they were divided into 2 groups(Group A (43 cases, 0°≤CC≤16.5°); Group B (35 cases, CC>16.5°)). The spinal drift distance, nerve recovery, axial symptoms and C5 palsy in the groups were recorded and analyzed. Results: The CC was 8.5°±3.8° in group A and 19.6°±3.0° in group B (t=14.071, P=0.000). The laminectomy width in group A was (22.1±1.7) mm, in group B was (21.8±1.5) mm, the difference between the two groups was not statistically significant (P>0.05). The distance of spinal cord backward movement was (1.7±0.4) mm in group A and (3.2±0.7) mm in group B. There was significant difference between the two groups (t=11.879, P<0.001). At the end of one year, the improvement rates of nerve function in the two groups were (63.3±13.1)% and (65.1±13.9)% respectively, there was no significant difference between the two groups (t=0.587, P=0.559). The VAS score of group A was (5.2±1.3) at one week and (3.5±0.6) at one month after operation, and that of group B was (3.8±0.8) and (2.4±0.4) respectively.There were statistically significant differences between groups at different time points (t=5.567, P<0.001; t=9.289, P<0.001). The incidence of C5 paralysis was 7.0% (3/43) in group A and 11.4% (4/35) in group B. There was no significant difference between the two groups (P>0.05). Conclusion The greater the curvature of cervical spine is, the more fully the spinal cord moves backward.The loss of curvature of cervical spine is related to the occurrence of axial symptoms.The curvature of cervical spine is not related to the recovery of nerve function and the occurrence of C5 nerve paralysis.

Objective To screen down regulated genes and find down regulated novel genes in gastric cancer. Methods Genes mRNA expression were detected between gastric cancer and normal gastric mucous membrane of five patients using cDNA microarray. Genes mRNA expression signals on hybridization membranes were analysized with computer software. Down regulated genes in gastric cancer were screened. cDNA suppression subtraction library was established by counterpart normal gastric mucous membrane mRNA(Tester) subtracting gastric cancer tissues mRNA(Driver) of five patients. After identification of the subtraction library, positive clones choosen randomly were sequenced , and down regulated novel genes in gastric cancer were screened. Some of the genes were identified by RT PCR.Results Down regulated genes in gastric cancer consist of 60 genes including tumor suppressing genes, apoptosis related genes, DNA replication and transcript or translate related genes, cell cycle related genes, cell migration related genes, etc. Two unknown gene fragments in gastric cancer were cloned. Conclusions Sixty down regulated genes in gastric cancer are confirmed. They are involved in gastric tumorigenicity and metastasis. cDNA subtraction library of gastric cancer was constructed successfully. Two down regulated novel gene fragments in gastric cancer was found.

OBJECTIVETo investigate the content and activity of M-phase promoting factor (MPF) in pleomorphic adenoma, mucoepidermoid carcinoma, buccal carcinoma and normal tissue, in order to evaluate the role of MPF in the development of tumor and the relationship between MPF and malignant degree.

METHODSThe content and activity of MPF were assessed by immunobloting and Gollicano method.

RESULTSThe cdc2 and cyclinB (two subunits of MPF) were found both in normal and tumor tissues, and their content in tumor was higher than normal tissues. Buccal carcinoma was 64% higher than normal tissues. The activity of MPF in carcinoma was higher than normal tissue and had positive relation with the malignant extent.

CONCLUSIONSThe content and activity of MPF in tumor are higher than normal tissue. PKC can activate MPF. These results show PKC may promote tumor proliferation by activating MPF and also, the activity of MPF has some relation with malignant extent.

CDC2 Protein Kinase ; analysis ; Cyclin B ; analysis ; Humans ; Immunoblotting ; Maturation-Promoting Factor ; analysis ; Mouth ; chemistry ; Mouth Neoplasms ; chemistry ; Protein Kinase C ; physiology

OBJECTIVETo study the regularity of the passive tensile of the masticatory muscles and ligaments by Twin-Block appliance under various bite reconstruction, and to provide some biomechanical references for the clinical use and improvement of Twin-Block appliance.

METHODS"Temporomandibular joint, mandible and Twin-Block appliance" model was set up by the three dimensional finite element method, and the related masticatory muscles and ligaments were added on it. Seven experimental groups were designed according to the clinical and research, the occlusal inclined plate's angles of Twin-Block appliance were 40 degrees, 45 degrees, 50 degrees, 55 degrees, 60 degrees, 65 degrees and 70 degrees. The passive tensile in the masticatory muscles and ligaments were analyzed by the computer.

RESULTSUnder various experimental groups, the passive tensile in the anterior deep masseter (AM), the posterior deep masseter (PM), the anterior temporalis (AT), the posterior temporalis (PT), the stylomandibular ligament and sphenomandibular ligament improved with the increased slant angles of occlusal guide. The maximum value of the passive tensile was 82.57 N, the minimum value was 0.07 N.

CONCLUSION1) In various experimental groups, AT, AM, PM, PT, stylomandibular ligament and sphenomandibular ligament are subject to passive tension force in the process of Twin-Block appliance guiding the mandibular forward and play the important role on the remodeling of the mandible. 2)All groups of occlusal inclined plate's angle are in physiologically tolerable range and can be used in clinic.

Dental Occlusion ; Humans ; Ligaments ; Mandible ; Masseter Muscle ; Masticatory Muscles ; Reconstructive Surgical Procedures ; Temporal Muscle ; Temporomandibular Joint

The purpose of this study is to reveal the role of Girdin in regulating the aggregation of actin filaments by studying the relationship between PKB/Akt and Girdin. First we used Scansite software (http://scansite.mit.edu) to predict relevant target sites of PKB/Akt on mouse Girdin. To gain insight into the role of phosphorylation of Girdin by PKB/Akt, we assessed the location of phosphorylated Girdin in fertilized eggs by staining with anti-P-Girdin 1 417 Ab. We detected a distinct increase in the fluorescence signal of F-actin and P-Girdin 1 417 after microinjection of Akt WT and myr-Akt. The addition of myr-Akt induced phosphorylation of Girdin in mouse fertilized eggs. In addition, siRNA-mediated Akt-knockdown blocked phosphorylation of Girdin. The distribution of actin filaments was obviously scattered. These results strongly suggest that PKB/Akt could directly phosphorylate Girdin on Ser1 417 and promote its function in mouse fertilized eggs.
Actins ; physiology ; Animals ; Mice ; Microfilament Proteins ; physiology ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; physiology ; RNA, Small Interfering ; Vesicular Transport Proteins ; physiology ; Zygote

ObjectiveTo explore the mechanism of Jiawei Wendantang in preventing and treating diabetic atherosclerosis by observing the effect of this prescription on the nuclear factor-κB / NOD-like receptor protein 3（NF-κB/NLRP3） pathway and related inflammatory cytokines in rat model of diabetic atherosclerosis. MethodFifty-four SPF-grade rats were randomized into blank， model， atorvastatin （0.9 mg·kg^-1·d^-1）， and high-， medium-， low-dose （18.2， 9.1， 4.55 g·kg^-1·d^-1， respectively） Jiawei Wendantang groups. The rats in other groups except the blank group were modeled for diabetic atherosclerosis by intraperitoneal injection of streptozotocin and feeding with a high-sugar high-fat diet， and those in the blank group were injected with an equal dose of citric acid buffer and fed with a regular diet. The drug administration lasted for 4 weeks， and the blood glucose level in the tail vein was measured every 6 days. After the last administration， the rats were anesthetized for sample collection. Enzyme-linked immunosorbent assay was employed to measure the serum levels of interleukin-18 （IL-18）， C-reactive protein （CRP）， tumor necrosis factor-α （TNF-α）， and intercellular adhesion molecule-1 （ICAM-1）. Western blot was employed to determine the relative protein levels of NF-κB p65， NLRP3， and ICAM-1 in the abdominal aorta. Real-time quantitative polymerase chain reaction was employed to determine the mRNA levels of NLRP3 and interleukin-1β （IL-1β） in the abdominal aorta. The pathological changes in the thoracic aorta were observed by hematoxylin-eosin staining. ResultCompared with the blank group， the model group showed elevated levels of IL-18， CRP， TNF-α， and ICAM-1 in the serum and blood glucose （P<0.05， P<0.01）， up-regulated protein levels of NF-κB p65， NLRP3， and ICAM-1 （P<0.01）， and up-regulated mRNA levels of NLRP3 and IL-1β （P<0.05）. Compared with model group， Jiawei Wendantang lowered the levels of IL-18， CRP， TNF-α， ICAM-1 and blood glucose （P<0.05， P<0.01）， down-regulated the protein levels of NF-κB p65， NLRP3， and ICAM-1 （P<0.01）， and down-regulated the mRNA levels of NLRP3 and IL-1β （P<0.05， P<0.01）. Moreover， Jiawei Wendantang alleviated the pathological injuries in the thoracic aorta. ConclusionJiawei Wendantang may modulate the NF-κB/NLRP3 signaling pathway to reduce the release and adhesion of inflammatory cytokines and regulate the blood glucose level to treat diabetic atherosclerosis.