AIMTo establish a method for identification of the activities of matrix metalloproteinases (MMPs) synthesized by cultured hepatic stellate cells (HSC).

METHODSSDS-gelatin polyacrylamide gels electrophoresis was used for the identification of MMPs synthesized by cultured HSC.

RESULTSMMP-2 and MMP-9 in culture medium and HSC were sensitively identified, and MMP-2 activity was higher than MMP-9 in culture medium, the changes of MMPs in culture medium and HSC were also observed after 24h medicine treatment.

CONCLUSIONZymography is suitable for the studies on the role of HSC in the regulation of extracellular matrix turnover.

Animals ; Cells, Cultured ; Electrophoresis, Polyacrylamide Gel ; methods ; Hepatic Stellate Cells ; metabolism ; Matrix Metalloproteinases ; metabolism ; Rats ; Rats, Wistar

OBJECTIVE: This study aims to assess the effects of the different thicknesses of body-shade resin layers on the color of polyetheretherketone (PEEK)-Crea.lign restorations. METHODS: Five PEEK specimens with the thickness of 0.6 mm were prepared. The color values of PEEK specimens were measured. Afterward, opaque-shade resin layers (0.1 mm) and body-shade resin layers (1.5 mm) were stacked with mold. The five specimens were evenly ground to a thickness of 1.4, 1.2, 1.0, 0.8, 0.6, 0.4, 0.2, and 0.0 mm in sequence. After grinding and ultrasonic cleaning, the color value was measured. RESULTS: With the constant thickness of PEEK and 0.1 mm thickness of opaque-shade resin layer, the L*, a*, and b* values all showed downward trend with the increased thickness of the body-shade resin layer (1.0-1.4 mm). With the constant thickness of PEEK and 0.1 mm thickness of opaque-shade resin layer, the color difference between the adjacent groups was less than 1.5 NBS. This difference between nonadjacent groups was more than 1.5 NBS when the thickness of the body-shade resin layer reached 0.6 mm. Color difference between PEEK-Crea.lign restoration and PEEK was more than 1.5 NBS. CONCLUSIONS The thickness change in the body-shade resin layers influence the color of the PEEK-Crea.lign restorations. Using A2 shade Crea.lign, opaque-shade resin layer thickness is 0.1 and 0.6 mm thickness of body-shade resin layer can produce color which clinically acceptable.
Color ; Composite Resins ; Ketones ; Polyethylene Glycols

OBJECTIVE: To observe the effect of acupuncture therapy based on "gut-brain axis" on clinical manifestations and gastrointestinal symptoms of children with autism spectrum disorder (ASD). METHODS: A total of 66 children with ASD were randomly divided into an observation group and a control group, 33 cases in each group. The control group was treated by routine rehabilitation training. On the basis of the control group, the observation group was treated with acupuncture based on "gut-brain axis", and the acupoints were Touwei (ST 8), Shenting (GV 24), Sishencong (EX-HN 1), Tianshu (ST 25), Zhongwan (CV 12), Zusanli (ST 36), etc. Both treatments were given once every other day, 3 times a week, 4 weeks as a course of treatment, consecutively for 3 courses. The scores of autism behavior checklist (ABC), TCM symptoms of gastrointestinal disease and childhood autism rating scale (CARS) were compared between the two groups before and after treatment, and the clinical efficacy was evaluated. RESULTS: After treatment, the scores of ABC, CARS and TCM symptoms of gastrointestinal disease in the two groups were lower than before treatment (P<0.05), and those in the observation group were lower than the control group (P<0.05). The total effective rate of the observation group was 90.9% (30/33), which was higher than 81.8% (27/33) in the control group (P<0.05). CONCLUSION On the basis of routine rehabilitation training, acupuncture therapy based on "gut-brain axis" is effective in treating ASD, which can relieve the clinical manifestations and gastrointestinal symptoms.
Child ; Humans ; Acupuncture Therapy ; Autism Spectrum Disorder/therapy* ; Gastrointestinal Diseases/therapy*

Objective To retrospectively analyze 40 cases of hanging and 33 cases of ligature strangulation in Kunming, to explore the neck injury characteristics and similarities and differences of related asphyxia signs of corpses in hanging and ligature strangulation cases, in order to provide reference for forensic identification. Methods Statistics of hanging and ligature strangulation cases accepted by Kunming Municipal Public Security Bureau from 2000 to 2017 were collected. Data including the gender, age, injury tool, neck injury and related asphyxia signs of the deceased in hanging and ligature strangulation cases were statistically tested by SPSS 23.0. Results There were more males in hanging cases than females. However, there were more females than males in ligature strangulation cases. In hanging cases, suicide was common, while homicide was rare. In ligature strangulation cases, homicide was common, while suicide or accidental death was rare. The average age of the deceased in hanging cases were older than those in ligature strangulation cases. The ligature mark in hanging was usually above the thyroid cartilage. The ligature mark in ligature strangulation was usually at the same level of the thyroid cartilage. The most common vital reactions were exfoliation and subcutaneous hemorrhage at the ligature mark, common among the deceased in hanging cases. Hyoid fracture rate of the deceased in ligature strangulation cases was more common than those in hanging cases. Conclusion The gender, age distribution, position of ligature mark, detection rate of vital reactions at the ligature mark and hyoid fracture rate of the deceased can help distinguish hanging from ligature strangulation.
Asphyxia ; Female ; Homicide ; Humans ; Male ; Neck Injuries ; Retrospective Studies ; Suicide ; Thyroid Cartilage

OBJECTIVE: To explore the method and curative effect of double bundle reconstruction of posterior cruciate ligament with total internal short tendon technique. METHODS: From October 2010 to June 2018, 50 patients with simple posterior cruciate ligament rupture were admitted, including 35 males and 15 females, ranging in age from 20 to 45 years old, 16 with acute injury and 34 with old injury. The posterior cruciate ligament (PCL) was reconstructed by double bundle technique with the whole internal short tendon. The two ends of the transplanted ligament were fixed by titanium plate suspended with adjustable loop. The Lysholm knee score and IKDC score were used to evaluate the clinical effects. RESULTS: All the 50 patients were followed up, and the duration ranged from 6 to 60 months, with a mean of (28.84±9.52) months. At the latest follow up, the knee joint activity returned to normal range. The median value of Lysholm knee score was 54 before operation and 100 after operation. According to the IKDC score, there were 0 case of grade A, 0 case of grade B, 20 cases of grade C and 30 cases of grade D before operation;29 cases of grade A, 19 cases of grade B, 1 case of grade C and 1 case of grade D after operation;the difference was statistically significant (<0.01). CONCLUSION Arthroscopic double bundle reconstruction of posterior cruciate ligament with short internal tendon is safe and reliable, with less tendon transplantation, more stable fixation, and more bone reserve, which is beneficial for healing and renovation. The short term effect is positive.
Adult ; Anterior Cruciate Ligament Injuries ; Arthroscopy ; Female ; Humans ; Joint Instability ; Knee Joint ; Male ; Middle Aged ; Posterior Cruciate Ligament ; Tendons ; Treatment Outcome ; Young Adult

OBJECTIVE: To investigate the therapeutic effect of Epothilone D on traumatic optic neuropathy (TON) in rats. METHODS: Forty-two SD rats were randomized to receive intraperitoneal injection of 1.0 mg/kg Epothilone D or DMSO (control) every 3 days until day 28, and rat models of TON were established on the second day after the first administration. On days 3, 7, and 28, examination of flash visual evoked potentials (FVEP), immunofluorescence staining and Western blotting were performed to examine the visual pathway features, number of retinal ganglion cells (RGCs), GAP43 expression level in damaged axons, and changes of Tau and pTau-396/404 in the retina and optic nerve. RESULTS: In Epothilone D treatment group, RGC loss rate was significantly decreased by 19.12% (P=0.032) on day 3 and by 22.67% (P=0.042) on day 28 as compared with the rats in the control group, but FVEP examination failed to show physiological improvement in the visual pathway on day 28 in terms of the relative latency of N2 wave (P=0.236) and relative amplitude attenuation of P2-N2 wave (P=0.441). The total Tau content in the retina of the treatment group was significantly increased compared with that in the control group on day 3 (P < 0.001), showing a consistent change with ptau-396/404 level. In the optic nerve axons, the total Tau level in the treatment group was significantly lower than that in the control group on day 7 (P=0.002), but the changes of the total Tau and pTau-396/404 level did not show an obvious correlation. Epothilone D induced persistent expression of GAP43 in the damaged axons, detectable even on day 28 of the experiment. CONCLUSION Epothilone D treatment can protect against TON in rats by promoting the survival of injured RGCs, enhancing Tau content in the surviving RGCs, reducing Tau accumulation in injured axons, and stimulating sustained regeneration of axons.
Animals ; Disease Models, Animal ; Epothilones ; Evoked Potentials, Visual ; Nerve Regeneration/physiology* ; Optic Nerve Injuries/metabolism* ; Rats ; Rats, Sprague-Dawley ; Retinal Ganglion Cells/physiology*

With the development of the computer simulation technology and the digital simulation technology, the traditional calculation method has been gradually replaced by the digital method to deal the road traffic accident scene and analyse the process. The PC-Crash software simulation system can reconstruct the traffic accidents within 32 vehicles, and the accuracy of reconstruction has been fully verified, which is widely used by the transport police department and the accreditation agency. In this paper, the research of road traffic accident reconstruction using PC-Crash software is reviewed, and the application of road traffic accident reconstruction technology based on PC-Crash software and some existing problems in forensic practice are discussed, which provides reference for the research and identification of road traffic accident simulation and reconstruction and theoretical basis for accident treatment.
Accidents, Traffic ; Computer Simulation ; Humans ; Models, Theoretical ; Police ; Software

Objective To investigate the effects of Tannic acid on the proliferation of human colon cancer SW 620 cell line and the mRNA and protein levels of TMEM16A .Methods Human colon cancer cell line SW620 were divided into the low dose(50 μmol/L) ,high dose(100 μmol/L) ,they were cultured for 48 h or 72 h separately .Control groups were cultured in the medium with DMSO .The proliferation of SW620 cell line was detected by the MTT assay at different time points (48 h or 72 h) .The cell cycle and apoptosis in the Tannic acid-treated groups were detected by flow cytometry .RT-PCR and Western blotting were used to de-termine the mRNA and protein levels of TMEM16A separately .All data were analyzed using the one-way analysis of variance (ANOVA) ,SNK test by the SPSS software .Results Compared with the control group ,the proliferation of SW620 cell line was significantly inhibited after the treatment by Tannic acid at the concentration of 50 μmol/L and 100 μmol/L for 48 h or 72 h(t=15 .35 ,P<0 .01 ;t=22 .32 ,P<0 .01) .Determined by flow cytometry ,after treated with Tannic acid ,the numbers of SW620 cells were inhibited in the G0/G1 phase and the S phase(F=6 782 .1 ,1 509 .3 ,P<0 .01) ,and the numbers of SW620 cells in G2/M phase were not varied in each group(F=1 .37 ,P>0 .05) ,and increased apoptotic rate when compared with control group (F=545 .3 ,P<0 .01) .The value of 3H-TdR and 3H-Leucine incorporation of SW620 cells treated with Tannic acid(100 μmol/L) 48 h and 72 h separately ,were obviously decreased as compared with that of control group (P<0 .05) .In the low dose treated groups (50 μmol/L) ,the mRNA levels in 48 h group and 72 h group were(0 .633 ± 0 .009) and(0 .621 ± 0 .011) ,and in the high dose treated groups (100 μmol/L) ,the mRNA levels in 48 h group(0 .64 ± 0 .15) and 72 h group(0 .63 ± 0 .11) ,were lower than the control group(F=7 .645 ,P< 0 .05) .After treating SW620 with Tannic acid for 48 h and 72 h ,in the low dose groups ,the protein expression of TMEM16A were(0 .68 ± 0 .14) and(0 .65 ± 0 .12) ,and in the high dose groups ,the protein expression of TMEM16A were(0 .64 ± 0 .15) and(0 .63 ± 0 .11) were decreased when compared with the control group (1 .28 ± 0 .06)(F=4 .508 ,P<0 .05) .Conclusion Tannic acid arrested SW620 at G1-S phase and decrease the mRNA and protein expression of TMEM16A .

Objective:To investigate whether the overexpression of Oct4B1 gene induces epithelial mesenchymal transition in human colorectal cancer SW480 cells and its possible mechanism.Methods: Experimental group(SW480-Oct4B1):Transfection of SW480 cell lines in colorectal cancer with Oct4B1 overexpression plasmid;Control group(SW480-Oct4B1):negative control plasmid with G418 resistance.Stably transfected cell lines were obtained by G418 culture medium.The two groups were compared with:①Detection of Oct4B1 gene expression in stably transfected cell lines by RT-qPCR;②Scratches and Transwell assays were used to estimate migration and invasion;③Detection of EMT related markers E-cadherin,N-cadherin and Vimentin protein expression by Western blot assay;④Detection of Twist gene and protein expression by RT-PCR and Western blot assays.Results: The transient transfection was confirmed by RT-qPCR and the stable transfected cell lines were obtained from two groups of cells transfected with G418 culture medium.Compared with the control group:①RT-qPCR revealed increased expression of Oct4B1 gene in the experimental group(P<0.01);②Cell migration and invasion were significantly increased(P<0.01);③Epithelial marker:the expression of E-cadherin protein was significantly decreased (P<0.01),interstitial marker:the expression of N-cadherin and Vimentin protein was significantly increased (P<0.01);④Twist mRNA and protein expression were significantly increased(P<0.01).Conclusion: Overexpression of Oct4B1 gene can induce epithelial mesenchymal transition in human colorectal cancer SW480 cells,its molecular mechanism may be related to the promotion of Twist expression.

Aim To compare the protective effects of gastrodin and melatonin on methamphetamine ( MA )-induced neurotoxicity of cortical neurons. Methods Cortical neuron cells were treated with different concentrations of methamphetamine for 24 h, then the cell viability was detected by CCK-8 kit to choose the optimal concentration of MA. The cells were treated with different concentrations of gastrodin or melatonin 2 h before the treatment of cells with optimal concentration of MA for 24 h. The optimal concentrations of gastrodin and melatonin were determined by CCK-8 kit as well. Cortical neuron cells were randomly divided into con trol group, MA group, gastrodin plus MA group, and melatonin plus MA group. Then the apoptotic rate of cells was detected by TUNEL method . The expression of Caspase-3 in cells was assessed by immunofluorescence. Results The optimal concentration of MA was 0. 5 mmol • L"1. After treating with MA, the numbers of cortical neurons decreased, the length of synapses became shorter, and the cell vacuolation, the number of dead cells increased. The optimal concentration of gastrodin was 25 mg • L'1, and the optimal concentration of melatonin was 10 jimol • L"1. Gastrodin treatment could reduce the apoptosis of cortical neurons induced by MA, and the expression of Caspase-3 decreased as well, and there was no significant difference compared with the intervention effect of gastrodin with melatonin. Conclusions MA has neurotoxic damage to cortical neurons. Gastrodin could attenuate MA-in-duced neurotoxicity via alleviating the apoptosis induced by MA.