1.Role of Caspase 3 in neuronal apoptosis after acute brain injury.
Xinyu YANG ; Shuyuan YANG ; Jianning ZHANG ; Liang XUE ; Zhen HU
Chinese Journal of Traumatology 2002;5(4):250-253
OBJECTIVETo analyze the role of Caspase 3 in neuronal apoptosis after acute brain injury.
METHODSExperiments were carried out with rat diffuse brain trauma model. The neuronal DNA injury in cortex and hippocampus was observed by TUNEL stain. The mRNA and protein expressions and enzyme activation of Caspase 3 were observed by Northern blot, in situ hybridization, immunohistochemistry stain and Western blot, respectively. Special Caspase 3 enzyme inhibitor was used to observe the therapeutic effect.
RESULTSTUNEL positive neurons appeared 2 hours after severe trauma, peaked at 1 day and lasted for 7 days. Northern blot showed that the Caspase 3 mRNA expression was increased and peaked at 1 day, about twice higher than the control. In the area of cortex and hippocampus, positive mRNA staining neurons appeared most distinct on one day. With the antibody for Caspase 3 P20 subunit, the active Caspase 3 expression peaked at 1-3 days. The electrophoresis band of PARP degradation would be seen by Western blot. Caspase 3 enzyme inhibitor could reduce apoptotic neuronal death without any effect on Caspase 3 P20 subunit expression.
CONCLUSIONSAfter brain trauma, Caspase 3 mRNA and protein expressions and enzyme activation are enhanced in combination with neuronal apoptosis. Special Caspase 3 enzyme inhibitor can apparently decrease the neuronal apoptosis.
Acute Disease ; Animals ; Apoptosis ; physiology ; Brain Injuries ; enzymology ; physiopathology ; Caspase 3 ; Caspases ; metabolism ; Enzyme Activation ; Enzyme Inhibitors ; pharmacology ; Nervous System ; physiopathology ; Neurons ; enzymology ; physiology ; RNA, Messenger ; metabolism ; Rats ; Rats, Wistar
2.Eye model for inspecting the functional vision of IOLs
Ren-Feng, XU ; Zhen-Ping, HUANG ; Hua-Chun, WANG ; De-Gao, ZHAO ; Liang, DING ; Chun-Yan, XUE ; Yong, WU
International Eye Science 2006;6(6):1233-1236
· AIM: To inspect and compare the functional vision of an aspheric intraocular lens (Tecnis) with those of conventional monofocal silicone and acrylic intraocular lens and multifocal intraocular lens (Array).· METHODS: The IOLs were tested in the eye model, which was designed to be optically equivalent to the theoretical eye model. The eye model is a combination of a spherical photographic lens with 35mm focal length ( IOL put in a water cell)and a charge coupled device (CCD) camera. The images constructed by the lenses are observed on a monitor of personal computer and the contrasts of the images are analyzed by using commercial image processing software.· RESULTS: The modulation transfer function of the eye model equals the scale produced by the theoretical eye model. The images constructed by changing the diameter of aperture stop and IOL.· CONCLUSION: The proposed eye model is useful for testing functional vision and for inspecting differences of intraocular lens.
3.The loss-of-function mutations and down-regulated expression of ASB3 gene promote the growth and metastasis of colorectal cancer cells
Lu ZHEN-HAI ; Zhou YI ; Kuang CHUN-MEI ; Wu LIANG-XUE
Chinese Journal of Cancer 2017;36(1):11-25
Backcground:Ankyrin repeat and SOCS box protein 3 (ASB3) is a member of ASB family and contains ankyrin repeat sequence and SOCS box domain.Previous studies indicated that it mediates the ubiquitination and degradation of tumor necrosis factor receptor 2 and is likely involved in inflammatory responses.However,its effects on oncogenesis are unclear.This study aimed to investigate the effects of ASB3 on the growth and metastasis of colorectal cancer (CRC).Methods:We used next-generation sequencing or Sanger sequencing to detect ASB3 mutations in CRC specimens or cell lines,and used real-time quantitative polymerase chain reaction,Western blotting,and immunohistochemical or immunofluorescence assay to determine gene expression.We evaluated cell proliferation by MTT and colony formation assays,tested cell cycle distribution by flow cytometry,and assessed cell migration and invasion by transwell and wound healing assays.We also performed nude mouse experiments to evaluate tumorigenicity and hepatic metastasis potential of tumor cells.Results:We found that ASB3 gene was frequently mutated (5.3%) and down-regulated (70.4%) in CRC cases.Knockdown of endogenous ASB3 expression promoted CRC cell proliferation,migration,and invasion in vitro and facilitated tumorigenicity and hepatic metastasis in vivo.Conversely,the ectopic overexpression of wild-type ASB3,but not that of ASB3 mutants that occurred in clinical CRC tissues,inhibited tumor growth and metastasis.Further analysis showed that ASB3 inhibited CRC metastasis likely by retarding epithelial-mesenchymal transition,which was characterized by the up-regulation of β-catenin and E-cadherin and the down-regulation of transcription factor 8,N-cadherin,and vimentin.Conclusion:ASB3 dysfunction resulted from gene mutations or down-regulated expression frequently exists in CRC and likely plays a key role in the pathogenesis and progression of CRC.
4.Etiological analysis and significance of anterior knee pain induced by gluteal muscles contracture.
Gang ZHAO ; Yu-jie LIU ; Jun-liang WANG ; Wei QI ; Feng QU ; Bang-tuo YUAN ; Jiang-tao WANG ; Xue-zhen SHEN ; Yang LIU ; Juan-li ZHU
China Journal of Orthopaedics and Traumatology 2014;27(12):1000-1002
OBJECTIVETo explore causes of gluteal muscle contracture induced anterior knee pain and curative effect of arthroscopic release.
METHODSFrom March 2002 to August 2013,36 patients with gluteal muscle contracture induced anterior knee pain were treated, including 15 males, 21 females, aged from 9 to 40 years old with an average (18.7±7.2) years old; the courses of diseases ranged from 4 to 30 years. The clinical manifestations involved limited to symmelia, positive Ober sign, buttocks touch contracture belts, knee and patella slide to lateral when doing squat activities. All patients were performed gluteal muscle contracture release under arthroscopic. Postoperative complications were observed, Kujala scoring before and after operation was used for compare curative effect.
RESULTSAll patients were followed up with an average of 29 months. The incision were healed well, and no complications were occurred. Postoperative Kujala score were improved more than preoperative.
CONCLUSIONGluteal muscle contracture release could alleviate hypertension of lateral patella, and palys an important role in preventing patellofemoral arthritis.
Adolescent ; Adult ; Buttocks ; Child ; Contracture ; physiopathology ; Female ; Humans ; Knee ; Male ; Pain ; etiology
5.Arthroscopic repair of meniscus injury with Fast-fix under local anesthesia.
Jiang-tao WANG ; Yu-jie LIU ; Jun-liang WANG ; Feng QU ; Bang-tuo YUAN ; Gang ZHAO ; Xue-zhen SHEN ; Juan-li ZHU ; Yang LIU
China Journal of Orthopaedics and Traumatology 2014;27(8):683-685
OBJECTIVETo evaluate the clinical outcome of arthroscopic repair method of meniscus injury with Fast-fix under local anesthesia.
METHODSFrom October 2005 to September 2012,106 patients with meniscus injuries admitted into our - hospital were studied, including 74 males and 32 females, ranging in age from 13 to 71 years old, averaged 27.6 years old. The duration of the disease ranged from 15 days to 5 years. The main clinical manifestations included knee joint pain after exercise, joint locking, pressing pain of knee joint and positive McMurray signs. The MRI showed meniscus tear or degeneration. Arthroscopic repairing surgeries were performed with Fast-fix under local anesthesia. Each patient was assessed with VAS pain evaluation and Lysholm knee-joint score system before and after operation. All the patients were followed up more than 1 year.
RESULTSOne hundred and 2 patients were followed up by recording subjective symptoms, clinical examinations and questions naires for an average of 2.6 years (ranged, 1.1 to 8 years), and 4 patients lost follow-up. All the 102 patients had no anesthetic complications. Ninety-six patients had normal subjective symptom and clinical examinations. Four patients had a mild ache with activities,2 patients had moderate pain after activities with joint space pressing pain. VAS pain evaluation and Lysholm knee-joint score after operation both were much better than that before operation.
CONCLUSIONonclusion: Local anesthesia can provide nice circumstances for surgeries. Arthroscopic repair using Fast-fix is an idea method for meniscus injury, especially for the posterior horn tear of medial meniscus, which is simple and convenient with less complications, and satisfactory results.
Adolescent ; Adult ; Aged ; Anesthesia, Local ; methods ; Arthroscopy ; methods ; Female ; Humans ; Knee Injuries ; surgery ; Magnetic Resonance Imaging ; Male ; Menisci, Tibial ; surgery ; Middle Aged ; Tibial Meniscus Injuries ; Visual Analog Scale ; Young Adult
6.Method for drainage of lymph fluid and determining the change of active materials in lymph fluid after rat ischemia-reperfusion injury.
Xue-Feng CHEN ; Gui-Zhen HE ; Liang-Guang DONG ; Hong SHU ; Xiu-Rong WANG ; Feng-Yan GAO
Acta Academiae Medicinae Sinicae 2009;31(3):322-325
OBJECTIVETo set up a method for the drainage of lymph fluid and explore the change of active materials in lymph fluid and serum after rat ischemia-reperfusion injury.
METHODSThe method of the drainage of lymph fluid was well established. Sixteen healthy male rats of SPF grade were selected and randomly divided into 2 groups: intestinal ischemia-reperfusion + drainage group (I/R + drainage group) and drainage group. All the rats were subjected to superior mesenteric artery occlusion for 60 minutes, followed by 120 minutes of reperfusion. We compared the change of high mobility group box-1 (HMGB1) protein, endotoxin tumor necrosis factor alpha (TNF-alpha), interleukin (IL) -1 beta, IL-6, and soluble vascular cell adhesion molecular-1 (sICAM-1) by draining lymph fluid and collecting serum in 2 groups.
RESULTSThe drainage of lymph fluid was successfully performed. The HMGB1, endotoxin, and cytokines in serum and lymph fluid were significantly higher in ischemia-reperfusion group than in drainage group (P < 0. 05).
CONCLUSIONSThe method for drainage of lymph fluid is simple and feasible. Endotoxin, HMGB1, and some cytokines in serum and lymph fluid may mediate the ischemia-reperfusion injury.
Animals ; Disease Models, Animal ; Drainage ; methods ; Endotoxins ; metabolism ; HMGB1 Protein ; metabolism ; Interleukin-6 ; metabolism ; Intestines ; blood supply ; metabolism ; Lymph ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism
7.Identification and analysis of Corydalis boweri, Meconopsis horridula and their close related species of the same genus by using ITS2 DNA barcode.
Rong-kun DOU ; Zhen-fei BI ; Rui-xue BAI ; Yao-yao REN ; Rui TAN ; Liang-ke SONG ; Di-qiang LI ; Can-quan MAO
China Journal of Chinese Materia Medica 2015;40(8):1453-1458
The study is aimed to ensure the quality and safety of medicinal plants by using ITS2 DNA barcode technology to identify Corydalis boweri, Meconopsis horridula and their close related species. The DNA of 13 herb samples including C. boweri and M. horridula from Lhasa of Tibet was extracted, ITS PCR were amplified and sequenced. Both assembled and web downloaded 71 ITS2 sequences were removed of 5. 8S and 28S. Multiple sequence alignment was completed and the intraspecific and interspecific genetic distances were calculated by MEGA 5.0, while the neighbor-joining phylogenetic trees were constructed. We also predicted the ITS2 secondary structure of C. boweri, M. horridula and their close related species. The results showed that ITS2 as DNA barcode was able to identify C. boweri, M. horridula as well as well as their close related species effectively. The established based on ITS2 barcode method provides the regular and safe detection technology for identification of C. boweri, M. horridula and their close related species, adulterants and counterfeits, in order to ensure their quality control, safe medication, reasonable development and utilization.
Base Sequence
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China
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Corydalis
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chemistry
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classification
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genetics
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DNA Barcoding, Taxonomic
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methods
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DNA, Plant
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chemistry
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genetics
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DNA, Ribosomal Spacer
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chemistry
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genetics
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Molecular Sequence Data
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Nucleic Acid Conformation
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Papaveraceae
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chemistry
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classification
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genetics
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Phylogeny
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Plants, Medicinal
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chemistry
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classification
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genetics
8.NR2B-pERK1/2-pElk-1 signaling contributes to the avoidance learning and memory of rats.
Xu-Hong CHEN ; Si-Yun SHU ; Zhen-Jiang LIANG ; Xin-Min BAO ; Li-Xue CHEN ; Yong-Ming WU
Chinese Journal of Applied Physiology 2007;23(1):121-125
AIMTo investigate whether NR2B-pERK1/2-pElk-1 signaling contributes to the Y-maze learning and memory of rat brain.
METHODS45 adult male SD rats were divided into 4 groups: (1) Ifenprodil peritoneal injection group (Ifenprodil ip, n = 14); (2) DMSO peritoneal injection group(DMSO ip, n = 15); (3) Ifenprodil cerebral ventricle injection group (Ifenprodil ic, n = 8); (4) DMSO cerebral ventricle injection group(DMSO ic, n = 8). Y-maze training and test were used as an learning and memory enhancing stimulus. Immunohistochemical and Western blotting methods were used for detecting pERK1/2 and pElk-1 expression intensity of different brain regions.
RESULTSCompared with the DMSO ip group, the ifenprodil ip group showed no change on the Y-maze learning score (P > 0.05), but its Y-maze memory score tested 24 after learning decreased (P < 0.05). Ifenprodil peritoneal injection made brain pERK1/2 and pElk-1 expression decreased generally. In hippocampus, marginal division of striatum(MrD), amygdala,these changes were more significant (P < 0.05). Compared with the DMSO ic group, the reconsolidation of Y-maze memory tested 6 hours after ifenprodil injection was impaired in ifenprodil ic group (P < 0.05). The OD value of pERK1/2 and pElk-1 positive bands in ifenprodil ic group attenuated generally. The pElk-1 positive bands of caudate putamen and MrD almost disappeared in ifenprodil ic group.
CONCLUSIONNR2B is essential for the formation of long-term memory, reconsolidation of Y-maze memory. The deactivation of NR2B by ifenprodil will impair these courses. Meanwhile, the deactivation of NR2B attenuates pERK1/2 and pElk-1 expression of learning and memory related regions after Y-maze learning and memory reconsolidation test. In MrD and caudate putamen, the pElk-1 expression are completely blocked by ifenprodil after memory reconsolidation test.
Animals ; Avoidance Learning ; physiology ; Dimethyl Sulfoxide ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Male ; Maze Learning ; physiology ; Memory ; physiology ; Piperidines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; metabolism ; ets-Domain Protein Elk-1 ; metabolism
9.The characteristics of proximal deep vein thrombosis after total knee and hip arthroplasty
Yao YAO ; Zhen RONG ; Long XUE ; Liang QIAO ; Xianfeng YANG ; Xingquan XU ; Kai SONG ; Xiaoyu DAI ; Yeshuai SHEN ; Dongyang CHEN ; Zhihong XU ; Dongquan SHI ; Jin DAI ; Qing JIANG
Chinese Journal of Orthopaedics 2017;37(6):360-367
Objective To analyze the characteristics of proximal deep vein thrombosis (DVT) after total joint arthroplasty.Methods 40 cases of proximal DVT were collected from May 2007 to October 2016 which were all diagnosed by lower limb venography or sonography.There were 9 males and 31 females aged from 38 to 86 years,average 67 years,and body mass index was from 16.94 to 31.25kg/m2,with an average of 24.3 kg/m2.29 cases of THA patients and 11 cases of TKA patients.The anatomical distribution,size,radiological performance,local physical findings were recorded and multivariable analysis was performed to evaluate the relationship between distribution of proximal DVT and related parameters including age,gender (male/female),side of leg(left/right),surgery type(knee and hip),preoperative diagnosis(femoral neck fracture/other disease),time of diagnosis with DVT (during hospitalization/after discharge).Results Among 40 proximal DVT cases,31 cases diagnosed by phlebography and 9 cases were diagnosed by ultrasound.2 cases were isolated proximal DVT while 38 cases connected with distal DVTs.9 cases of blood clots in the proximal and distal thrombosis connected discontinuously.There was no isolated proximal DVT in TKA group and 10 cases (90.9%) of blood clots in the proximal and distal thrombosis connected continuously while there were 2 cases (6.9%) isolated proximal DVTs in THA group and 21 (77.8%) of blood clots in the proximal and distal thrombosis connected discontinuously,and there was no statistic significant difference between the two groups.The average length of proximal was 8.85±9.3 1cm (range from 2-35cm) with 5.0±2.05 cm in TKA group and 10.31 ± 10.55 cmin THA group.A significant difference was found between the 2 groups (P=0.014).13 cases located in femoral veins and upper venous region which were all from THA group.There was a significant difference in the distribution between surgery type and preoperative diagnosis.However,there was no significant difference between other potential factors and distribution of proximal DVT.25 cases accepted the evaluation of symptom of DVT before scanning.However,no significant difference were found in edema,VAS score,Homans sign,Neuof sign between 9 TKA and 16 THA cases.Conclusion Proximal DVTs after knee and hip arthroplasty are more preferred to connect with distal DVT continuously.Surgery type and preoperative diagnosis have a significant effect on the distribution of proximal DVT.Proximal DVTs in THA and patients with preoperative femoral neck fracture could be more easily involving femoral veins and upper parts.
10.DADLE suppresses the proliferation of human liver cancer HepG2 cells by activation of PKC pathway and elevates the sensitivity to cis-diammine dichloridoplatium.
Bo TANG ; Jian DU ; Zhen-ming GAO ; Rui LIANG ; De-guang SUN ; Xue-li JIN ; Li-ming WANG
Chinese Journal of Oncology 2012;34(6):425-429
OBJECTIVETo investigate the effect of DADLE, a δ-opioid receptor agonist, on the proliferation of human liver cancer HepG2 cells and explore the mechanism involving PKC pathway.
METHODSHepG2 cells were treated with DADLE at different doses (0.01, 0.1, 1.0 and 10 µmol/L). Cell viability was determined using methyl thiazolyl terazolium (MTT) assay. The expression of PKC mRNA and p-PKC protein were examined by RT-PCR and Western blot assay. After treated separately with DADLE plusing NAL or PMA, the cell cycle of HepG2 cells was analyzed by flow cytometer. MTT was used to detect their proliferation capacity and Western blot was used to examine the p-PKC expression. The growth inhibitory rate of HepG2 cells treated with DADLE and cis-diammine dichloridoplatinum (CDDP) was analyzed.
RESULTSDADLE at different concentrations showed an inhibitory effect on the proliferation of HepG2 cells though inhibiting the expression of PKC mRNA and p-PKC protein. The results of flow cytometry showed that compared with the control group, the percentage of S + G(2)/M cells in DADLE-treated group was lowered by 3.94% (P < 0.01). Meanwhile, after treated with NAL and PMA, the percentage was elevated by 3.22% and 3.63%, respectively (P < 0.01). The MTT and Western blot assays showed that compared with the control group, the values of A570 and p-PKC protein levels in the HepG2 cells of DADLE-treated group were significantly decreased (P < 0.01). After treatment with NAL and PMA, the values of A570 and p-PKC protein levels were elevated significantly (P < 0.01). The growth inhibitory rate of DADLE + CDDP group was 79.9%, significantly lower than 25.2% and 43.2% of the DADLE and CDDP groups, respectively.
CONCLUSIONSActivation of δ-opioid receptor by DADLE inhibits the apoptosis of human liver cancer HepG2 cells. The underlying mechanism may be correlated with PKC pathway. DADLE can enhance the chemosensitivity of HepG2 cells to CDDP.
Antineoplastic Agents ; pharmacology ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cisplatin ; pharmacology ; Dose-Response Relationship, Drug ; Drug Resistance, Neoplasm ; Enkephalin, Leucine-2-Alanine ; administration & dosage ; pharmacology ; Hep G2 Cells ; Humans ; Naltrexone ; analogs & derivatives ; pharmacology ; Phosphorylation ; Protein Kinase C ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Receptors, Opioid, delta ; agonists ; Signal Transduction ; Tetradecanoylphorbol Acetate ; analogs & derivatives ; pharmacology