OBJECTIVETo explore the prevalence, distribution and clinical significance of high-intensity zone (HIZ) in anterior annulus fibrosus (AF) in comparison with HIZ in posterior AF of lumbar disc.

METHODSAccording to the diagnosis and location of HIZ, 610 lumbar magnetic resonance images with entire clinical materials were divided into control group (without HIZ), anterior AF group (HIZ), posterior AF group (HIZ) and anterior & posterior (AP) AF group (HIZ). The incidence of HIZ was summarized. The clinical data, such as male/female ratio, age, and body weight, prevalence of low back pain (LBP) and distribution of HIZ, were compared and analyzed between the groups.

RESULTSThree hundred and fifteen cases shown no HIZ (51.6%), 95 cases presented HIZ in anterior AF (15.6%, 119 discs), 159 cases presented HIZ in posterior AF (26.1%, 189 discs) and 41 cases presented HIZs in both anterior and posterior AF (6.7%, 96 discs). There was significant difference between the prevalence of HIZ in anterior AF and that in posterior AF (P < 0.01). The male/female ratio and body weight of each groups showed no difference (P > 0.05), and the age was proved to be statistically different between four groups (P < 0.01, control group < posterior AF group < AP AF group < anterior AF group). HIZs in anterior AF often occurred at L(1,2)-L(4,5), whereas, they usually developed at L(3,4)-L(5)S(1) in posterior AF. The incidence of LBP in control group, anterior AF group, posterior AF group, AP AF group were 40.0%, 52.6%, 55.4% and 65.8%, respectively. The LBP prevalence of control group was lower than that of other three groups (P < 0.05), and the prevalence of last three groups showed no difference (P > 0.05).

CONCLUSIONSCompared with HIZ in posterior AF, the HIZ in anterior AF of lumbar disc has a lower prevalence, often develops in elder patients and in upper motion segments. It also indicates an obvious relation to LBP as the former.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Intervertebral Disc ; pathology ; Intervertebral Disc Displacement ; diagnosis ; pathology ; Lumbar Vertebrae ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Young Adult

OBJECTIVETo construct the delta-pIRES2-EGFP plasmid and investigate its expression in HEK293 cells.

METHODSFull length cDNA of rat delta opioid receptor gene amplified from rat brain tissues using reverse transcription and nested PCR was cloned into pMD20 T vector. The delta cDNA was inserted into pIRES2-EGFP plasmid to construct the recombinant eukaryotic plasmid delta-pIRES2-EGFP, which was transfected into HEK293 cells via Lipofectamine2000. The expression of delta was examined under fluorescence microscope.

RESULTSThe recombinant delta-pIRES2-EGFP plasmid was successfully constructed, and high expression of delta was detected in HEK293 cells transfected by the plasmid.

CONCLUSIONdelta-pIRES2-EGFP has been successfully cloned, which shows high expression of delta in HEK293 cells.

Animals ; DNA, Complementary ; genetics ; Gene Expression ; Genetic Vectors ; Green Fluorescent Proteins ; biosynthesis ; genetics ; HEK293 Cells ; Humans ; Plasmids ; Polymerase Chain Reaction ; Rats ; Rats, Sprague-Dawley ; Receptors, Opioid, delta ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transfection

OBJECTIVETo investigate the protective effect of mild hypothermia on rat astrocytes with traumatic or ischemic injury.

METHODSRat astrocytes in primary culture were subjected to scratching or hypoxic injury and exposed to normothermia (37 degrees celsius;) or hypothermia (34 or 32 degrees celsius;) for 24 h. The morphology of the astrocytes was evaluated by live/dead staining, and the cell injury was measured by lactate dehydrogenase (LDH) release assay.

RESULTSAs the temperature reduced the LDH release rate from the cells in hypoxic group decreased significantly, to (11.48 - or + 1.53)% at 34 degrees celsius; and (3.79 - or + 0.45)% at 32 degrees celsius; as compared to that in normothermia [(33.02 - or + 3.58)%] in the absence of rat white blood cells (WBC) (P<0.001). LDH release rate of the hypoxic cells further decreased in the presence of rat WBC to (51.14 - or + 2.17 )% at 37 degrees celsius;, (19.53 - or + 4.37)% at 34 degrees celsius; and (16.68 - or + 1.47)% at 32 degrees celsius; (P<0.001). In the scratched cells, with or without WBC, LDH release rate showed no significant variation between the 3 temperatures (P>0.05).

CONCLUSIONMild hypothermia offers obvious protective effects on rat astrocytes against ischemic damage but not against mechanical injury.

Animals ; Animals, Newborn ; Astrocytes ; enzymology ; pathology ; Brain Injuries ; therapy ; Brain Ischemia ; therapy ; Cell Hypoxia ; Cells, Cultured ; Cold Temperature ; L-Lactate Dehydrogenase ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the effect of quercetin on the proliferation of neural stem cells in the subventricular zone (SVZ) of rats after focal cerebral ischemia.

METHODSAn adult rat model of middle cerebral artery occlusion (MCAO) model was established by placement of an intraluminal filament at the origin of the MCA. Quercetin was administered intraperitoneally in the rats at a dose of 50 mg/kg every 3 days starting at 6 h after MCAO, and BrdU (50 mg/kg daily) was also injected intraperitoneally starting at 4 h after MCAO. BrdU-positive cells in the SVZ were counted at 7, 14 and 21 days after MCAO.

RESULTSCompared with the sham-operated group, the rats in the ischemic model group showed significantly increased BrdU-positive cells in the ipsilateral SVZ 7 days after MCAO, reaching the peak level on day 14 and beginning to decrease on day 21 (P<0.05). The number of ipsilateral BrdU-positive cells in quercetin group was significantly greater than that in the model group on days 7, 14 and 21 (P<0.05), and maintained the high level on day 21.

CONCLUSIONQuercetin can maintain a high level of neural stem cell proliferation in the SVZ after focal cerebral ischemia in adult rats.

Animals ; Brain Ischemia ; pathology ; physiopathology ; Cell Proliferation ; drug effects ; Cerebral Ventricles ; pathology ; Infarction, Middle Cerebral Artery ; pathology ; physiopathology ; Male ; Neural Stem Cells ; cytology ; Quercetin ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; pathology

Objective To investigate the feasibility of microsurgery therapy with Toth water dissection technique for patients with hypertensive intracerebral hemorrhage via lateral fissure approach.Methods Forty-eight patients with hypertension basal ganglia cerebral hemorrhage,admitted to our hospital from January 2009 to January 2012,were selected in our study; they were divided into two groups according to different treatment methods:the observation group (n=24) chose Toth water dissection technique via lateral fissure approach and the control group (n=24) adopted surgery via lateral fissure approach.The operation time,postoperative coma time,hematoma clear rate,postoperative bleeding and activity of daily living (ADL) were compared between the two groups.Results Better results of operating time,postoperative coma time and hematoma clear rate in the observation group were noted as compared with those in the control group with significant differences (P＜0.05).All patients were followed up for 3 months; significant difference on ADL grading was noted between the observation group and the control group (Z=-1.998,P=0.046).The mean rank in the observation group (20.67) was obviously smaller than that in the control group (28.33),indicating that the prognosis in the observation group was better than that in the control group.No death was noted in the observation group while two in the control group.Conclusion Toth water dissection technique has better effect,enjoying smaller trauma,simpler operation,less time consuming and better protective effects on frontal and temporal lobes,in patients with hypertensive intracerebral hemorrhage as compared with traditional lateral fissure technology.

Objective To study the effect of quercetin on gene expression in glucose-oxygen deprived astrocytes using large-scale oligo microarray technology.Methods Astrocytes were primarily cultured in vitro and divided into ischemia and hypoxia group and ischemia and hypoxia plus quercetin treatment(50 μmol/L)group; ischemia and hypoxia cells from these 2 groups were induced by anaerobic culture for 4 h,and then,the nutrient solutions were added into each group,respectively.Twenty-four h after that,cDNA microarray was employed to select the differentially expressed genes in the 2 groups,and these genes were confirmed by quantitative RT-PCR.Results The cDNA microarray indicated that the expressions of 180 genes had significant changes at the mRNA level between ischernia and hypoxia group and ischemia and hypoxia plus quercetin treatment group,of which 49 genes were up-regulated and 131 were down-regulated.One hundred and forty-eight differentially expressed genes were confirmed by RT-PCR,including 34 up-regulated genes and 114 down-regulated genes,which showed that 82.2％(148/180)genes that matched with the results of cDNA microarray.Conclusion Gene expression profiling by large-scale oligo microarray provides good understanding of the molecular mechanism of quercetin in glucose-oxygen deprived astrocytes,and laids the foundation for investigating the influence of quercetin and astrocytes in hypoxic-ischemic brain damage.

OBJECTIVETo evaluate the effect of Zhuanggu Jianxi Decoction (, ZGJXD) on interleukin-1 β (IL-1 β)-induced degeneration of chondrocytes (CDs) as well as the activation of caveolin-p38 mitogen-activated protein kinase (MAPK) signal pathway, investigating the possible molecular mechanism that ZGJXD treats osteoarthritis.

METHODSSerum pharmacology was applied in the present study, where ZGJXD was orally administrated to New Zealand rabbits and then ZGJXD containing serum (ZGJXD-S) was collected for following in vitro experiments. CDs were isolated aseptically from New Zealand rabbits and then cultured in vitro. Upon IL-1 β stimulation, the degeneration of CDs was verified by inverted microscope, toluidine blue stain and type II collagen immunocytochemistry. After IL-1 β-stimulated CDs were intervened with blank control serum, ZGJXD-S, together with or without SB203580 (a specific inhibitor of p38 MAPK) for 48 h, caveolin-1 protein expression and the phosphorylation level of p38 were determined by Western blotting, and the mRNA expression of IL-1 β, tumor necrosis factor α (TNF-α), matrix metalloproteinase 3 (MMP-3) and MMP-13 were examined by real-time polymerase chain reaction.

RESULTSIL-1 β stimulation induced degeneration of CDs, increased caveolin-1 expression and p38 phosphorylation, up-regulated the mRNA level of IL-1 β, TNF-α, MMP-3 and MMP-13. However, the IL-1 β-induced activation of caveolin-p38 signaling and alteration in the expression of p38 downstream target genes were suppressed by ZGJXD-S and/or SB203580 in CDs.

CONCLUSIONZGJXD can prevent CDs degeneration via inhibition of caveolin-p38 MAPK signal pathway, which might be one of the mechanisms that ZGJXD treats osteoarthritis.

Animals ; Base Sequence ; Blotting, Western ; Caveolins ; metabolism ; Chondrocytes ; drug effects ; enzymology ; metabolism ; DNA Primers ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Profiling ; Interleukin-1beta ; physiology ; MAP Kinase Signaling System ; Male ; RNA, Messenger ; genetics ; Rabbits ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism

OBJECTIVETo analysis the clinic and genotype in two Chinese patients with Dyskeratosis congenita (DC).

METHODSThe two patients were characterized by mucocutaneous abnormalities (abnormal nails, lacey reticular pigmentation, and oral leukoplakia), bone marrow failure. They were diagnosed with DC. DC genes were amplified by polymerase chain reaction (PCR), including DKC1, TERT, TERC, TINF2, NOP10, NHP2, then DNA sequencing was performed for abnormal exons.

RESULTSAn abnormal peak was found in exon 6 of TINF2 gene of the two patients. DNA sequencing showed a 845G→A transition in TINF2 gene in the two patients.

CONCLUSIONWe should think about DC if the young patients with mucocutaneous abnormalities and marrow failure. TINF2 c.845G→A(R282H) does exist in the two patients. It is reported in China for the first time.

Base Sequence ; Child, Preschool ; DNA Mutational Analysis ; Dyskeratosis Congenita ; diagnosis ; genetics ; Exons ; Female ; Humans ; Infant ; Male ; Telomere-Binding Proteins ; genetics

OBJECTIVETo establish a novel model of lumbar disc degeneration on the early stage in the rhesus monkey using percutaneous needle puncture guided by CT.

METHODS(1) Thirteen rhesus monkeys aged from 4 to 7 years, female 7 and male 6 were selected for establishing a model of the early stage of lumbar disc degeneration. (2)13 monkeys, 91 discs were divided into 3 groups: 64 discs from L1/2 to L5/6 were percutaneous punctured with a needle 20G as experimental group and 1 disc with a needle 15G as puncture control group and 26 discs were not be punctured from L6,7 to L7-S1 as control group. (3) Lumbar disc localization for needle puncture was guided by CT. All discs were examined by MRI, the HE, Masson's trichrome, Safranine-O and immunohistochemical staining of type II collagen before disc puncture and after puncture at 4, 8 and 12 weeks.

RESULTSMRI: (1) Experimental group: Pfirmann's Grade I was shown at postoperation 4, 8 and 12 weeks; (2) Puncture control group: Grade III was shown at postoperation 4 weeks and Grade IV at 8 weeks; (3) CONTROL GROUP: Grade I was shown at postoperation 4, 8 and 12 weeks. Histological examination: (1) In experimental group, there was no any change at postoperation 4 weeks, and the cell population of the nucleus was decreased at 8 weeks and more decreased at 12 weeks in HE. (2) There was no any change at postoperation 4 weeks, the clefts among the lamellae of the annulus fibrosus (AF) were shown at 8 weeks and more wider of the clefts of AF at 12 weeks in Masson's trichrome. (3) No any change was shown at postoperation 4 weeks, proteoglycan were progressively decreased at 8 and 12 weeks in Safranine-O. (4) No statistically significant difference in positive rate was observed at 4 and 8 weeks compared with control group in immunohistochemical staining of type II collagen. There was statistical difference at 12 weeks compared with control group (P<0.05). In puncture control group postoperation 8 weeks, the morphology of cell of nucleus pulposus was not clear in HE. The wider clefts of lamellae of the AF were shown in Masson's trichrome. The proteoglycan was obviously decreased in Safranine-O. Immunohistochemical staining collagen II synthesized was decreased. In normal control group, no any change was shown at 4, 8 and 12 weeks.

CONCLUSIONSThe degeneration of lumbar intervertebral disc on the early stage could be induced by the percutaneous needle puncture (20G) to the annulus fibrosus. The assessment of disc degeneration on early stage is not shown on MRI and only confirmed by histological examination.

Animals ; Disease Models, Animal ; Female ; Intervertebral Disc ; metabolism ; pathology ; surgery ; Intervertebral Disc Displacement ; etiology ; metabolism ; pathology ; Lumbar Vertebrae ; surgery ; Macaca mulatta ; Male ; Minimally Invasive Surgical Procedures ; Random Allocation

OBJECTIVETo investigate the role of allograft inflammatory factor-1 (AIF-1) in colorectal cancer (CRC) progression and explore the possible mechanism.

METHODSThe expression levels of AIF-1 in 70 CRC tissues and paired adjacent tissues were detected using immunohistochemistry and Western blotting, and the correlation of AIF-1 expression with the clinicopathological features of the patients was analyzed. In the CRC cell line SW480, the functional role of AIF-1 in regulating tumor progression was investigated by transfecting the cells with an AIF-1-overexpressing plasmid (AIF-1) and a negative control plasmid (NC). EdU proliferation assay and flow cytometry were used to assess the cell proliferation and cell cycle changes; Transwell migration assay and Annexin V-APC/7-AAD apoptosis assay kit were used to analyze the cell migration and apoptosis. The changes in the biological behaviors of the cells were observed after application of SB203580 to block the p38 MAPK pathway. The expression levels of CDK4, cyclin D1, P21, P27, MMP2, MMP9, Bax, Bcl2, Bcl-xl, p38 and p-p38 were detected using Western blotting.

RESULTSAIF-1 was down-regulated in CRC tissues compared with the adjacent normal tissues, and its expression level was positively correlated with lymph node metastasis (P=0.008), TNM stage (P=0.003) and tumor size (P=0.023). Overexpression of AIF-1 in SW480 cells significantly reduced EdU-positive cells and caused obvious cell cycle arrest in G1 phase (P<0.05). AIF-1 overexpression resulted in significantly lowered protein expressions of CDK4 and cyclin D1, enhanced expressions of P21 and P27, attenuated cell migration ability (P<0.001), and decreased protein levels of MMP2 and MMP9. AIF-1 overexpression also induced obvious apoptosis of SW480 cells (P<0.01), significantly increased the protein levels of Bax and p-p38, and decreased the protein levels of Bcl-2 and Bcl-xl; SB203580 significantly attenuated the apoptosis-inducing effect of AIF-1 overexpression.

CONCLUSIONAIF-1 plays the role of a tumor suppressor in CRC by inhibiting cell proliferation, suppressing cell migration and inducing cell apoptosis. AIF-1 overexpression promotes the apoptosis of CRC cells by activating the p38 MAPK pathway.