Objective To investigate the relationship between the expression of matrix metalloproteinase-9 (MMP-9) and intercellular adhesion molecule-1 (ICAM-1) in the perihematomal tissues in human hypertensive,intracerebral hemorrhage (ICH) and brain edema formation following ICH.Methods Paraffin-embedded brain tissues of 39 human fatal cases of ICH from the perihematomal tissues,1—3 cm away from the margin of the hemorrhagic lesion,as well as tissues from the corresponding area at the opposite side as controls,were stained with HE and immunohistochemistry staining.The expressions of MMP-9 and ICAM-1 in the pefihematomal tissues were analyzed with the SPSS 11.5 system.Results ①With MMP-9 immunohistochemical staining positive capillaries in the perihematomal tissues were identified at 2 h ((1.2? 0.8)/HP).The number of MMP-9 positive capillaries began to rise at 5—10 h ((4.1?0.8)/HP) reaching the peak at 45—48 h ((10.6?1.4)/HP,P

OBJECTIVEPosterolateral intertransverse process fusion was performed in aged and young adult female rabbits lumbar spine using recombinant human bone morphogenetic protein-2 (rhBMP-2) and autograft to reveal the function of rhBMP-2 on spinal fusion on aged animals.

METHODSA total of 24 female New Zealand white rabbits included 12 young adult of 6 months and 12 aged of 2-year-old, was divided into 4 groups: (1) young adult autologous iliac crest bone group [ICBG(Y), n=6); (2) young adult rhBMP-2/absorbable collagen sponge (ACS) group [BMP-2(Y), n=6]; (3) aged autologous iliac crest bone group [ICBG(O), n=6]; aged rhBMP-2/ACS group [BMP-2(O), n=6]. All were underwent posterolateral fusion in same day. rhBMP-2 and autologous iliac crest bone was implant bilateral LS-L6 intertransverse processes, respectively. Half of the rabbits were sacrificed at 3.6 weeks following surgery, respectively. The results were assessed by manual palpation, radiographs, computed tomographic scans (3D) and histology.

RESULTSSix weeks after surgery, radiography, computed tomography and histology indicated the different result in healing in the posterolateral fusion using rhBMP-2 compared to ICBG (P < 0.05). Aged BMP-2 group showed significantly higher fusion rates than Aged ICBG group.

CONCLUSIONThis study demonstrated rhBMP-2 can increase the posterolateral fusion rate and new bone quality in aged rabbitss than autograft, it may take the place of ICBG. But its role is effected by age.

Aging ; Animals ; Bone Morphogenetic Protein 2 ; pharmacology ; Female ; Humans ; Palpation ; Rabbits ; Recombinant Proteins ; pharmacology ; Spinal Cord ; drug effects ; pathology ; surgery ; transplantation ; Spinal Fusion ; Tomography, X-Ray Computed ; Transplantation, Autologous

OBJECTIVETo investigate the in vitro effect of caffeic acid phenethyl ester (CAPE), a NF-κB inhibitor, on the apoptosis of osteoarthritic (OA) chondrocytes and on the regulation of the gelatinases matrix metalloproteinase 2 (MMP-2) and matrix metalloproteinase 9 (MMP-9).

METHODSAnnexin V-FITC/propidium iodide (PI) labeling and western blotting were used to observe and determine the apoptosis in TNFα-stimulated primary cultured osteoarthritic chondrocytes. Also, gelatin zymography was applied to examine MMP-2 and MMP-9 activities in supernatants.

RESULTSIt was confirmed by both flow cytometry and western blotting that chondrocytes from OA patients have an apoptotic background. Use of CAPE in combination with 10 ng/mL of TNFα for 24 h facilitated the apoptosis. MMP-9 in the supernatant could be autoactivated (from proMMP-9 to active MMP-9), and the physiologic calcium concentration (2.5 mmol/L) could delay the autoactivation of MMP-9. The activities of MMP-2 and MMP-9 in the fresh supernatant increased significantly in response to stimulation by 10 ng/mL of TNFα for 24 h. The stimulatory effect of TNFα just on proMMP-9 was counteracted significantly by CAPE.

CONCLUSIONNF-κB could prevent chondrocytes apoptosis though its activation was attributed to the increase of proMMP-9 activity induced by TNFα (a pro-apoptotic factor). Therefore, therapeutic NF-κB inhibitor was a 'double-edged swords' to the apoptosis of chondrocytes and the secretion of MMP-9.

Aged ; Apoptosis ; drug effects ; Caffeic Acids ; pharmacology ; therapeutic use ; Calcium ; physiology ; Cells, Cultured ; Chondrocytes ; drug effects ; enzymology ; secretion ; Drug Evaluation, Preclinical ; Female ; Humans ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Middle Aged ; NF-kappa B ; antagonists & inhibitors ; Osteoarthritis ; drug therapy ; enzymology ; Phenylethyl Alcohol ; analogs & derivatives ; pharmacology ; therapeutic use ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo establish a method for fingerprinting of Fuzhisan (FZS, a traditional Chinese medicinal preparation) using high-performance liquid chromatography with ultraviolet and evaporative light scattering detector (HPLC-UV/ELSD) to allow simultaneous determination of 5 major constituents in the preparation.

METHODSHPLC-UV/ELSD analysis was performed on water AlltechC18 column (5 microm, 4.6 mm x 250 mm) with a mixture of acetonitrile (A) and 0.1% acetice acid water (B) as the mobile phase. The solvent A gradient for elution was 0, 12%; 25, 20%; 30, 20%; 75, 30%; 105, 40%; 120, 80%; 130, 12%, with the flow rate of 1.0 ml/min; and the column temperature at 30 degrees . The detective wavelength was 335 nm, drift tube temperature was 80 degrees , pressure of nebulizer gas was 25 psi. The similarities between the HPLC-UV/ELSD fingerprints of the 12 extracts were calculated using similarity evaluation software.

RESULTSThe fingerprint of FZS was established and the 5 major constituents were identified. The complementarity between the fingerprints of UV and ELSD was analyzed, showing good correlation between 12 batches of FZS.

CONCLUSIONThe method for fingerprinting can simultaneously characterize the main chemical constituents in FZS and allows stable, effective and comprehensive quality control and evaluation of FZS for a single sample.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; standards ; Light ; Quality Control ; Scattering, Radiation ; Ultraviolet Rays

Objective To explore the characteristics and components of recurrent papillary thyroid cancer in patients with metabolic syndrome .Methods We retrospectively reviewed the information of the patients with recurrent papillary thyroid cancer after initial surgery during January 1st ,2010 and December 31st ,2015 in the First Affiliated Hospital of Zhengzhou University .We compared tumor size ,lymph node metastasis ,recurrence time and postoperative invasion rate in metabolic syndrome and non-metabolic syndrome groups . Results Totally 82 patients were included with 34 men and 48 women .There was no significant difference between patients with and those without metabolic syndrome grouped by the classic diagnosis approach .However ,the lymph node metastasis grade of recurrent papillary thyroid cancer patients who also suffered from at least two metabolic disorders ,was lower ,especially in women (P=0 .002) .Moreover ,patients with metabolic disorders had shorter recurrence time (Pdiabetes=0 .034 , Pdyslipidemia =0 .037 , PBMI =0 .004 , PMetS2 =0 .036) .Conclusion Papillary thyroid cancer patients with metabolic disorder ,especially with two or more components of metabolic syndrome and overweight and/or obesity ,may have an increasing risk of recurrence .

OBJECTIVETo clone the human tissue factor pathway inhibitor-2 (hTFPI-2) gene and express it by using prokaryotic expression system.

METHODSThe hTFPI-2 coding region was obtained by RT-PCR from human placenta total RNA. The coding fragment was then inserted into prokaryotic expression vector pET19b and expressed in E. coli BL21 by IPTG induction. The produced inclusion bodies were dissolved by denaturalizing chemicals, purified by ion exchange chromatograph, and refolded in air to form proper disulfide bonds. Chromogenic and gelatin zymography methods were used to evaluate the inhibiting effects of hTFPI-2 on trypsin, plasmin and MMPs individually. The inhibitory activity of hTFPI-2 on fabrisarcoma was investigated by matrigel.

RESULTSThe coding fragment of hTFPI-2 was cloned successfully and the protein was expressed as inclusion bodies which account for 20% - 30% of total host protein. The refolded hTFPI-2 could inhibit the invasive ability of fibrisarcoma HT-1080 as well as activity of plasmin, trypsin and MMPs.

CONCLUSIONSThe activated hTFPI-2 is obtained by using prokaryotic expressed system effectively.

Cloning, Molecular ; Escherichia coli ; metabolism ; Gene Expression ; Glycoproteins ; biosynthesis ; genetics ; Humans ; Placenta ; cytology ; RNA ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo explore the way to induce mesenchymal stem cells (MSCs) to differentiate into dopaminergic neurons in vitro.

METHODSMSCs were obtained from rat bone marrow, cultured and passaged. MSCs used in this experiment had multipotency, which was indirectly proved by being induced to differentiate into chondrocytes and adipocytes. MSCs were cultured in medium containing 0.5 mmol/L IBMX for 2 days. Then the medium was replaced with induction medium, which contained GDNF, IL-1beta, mesencephalic glial-cell-conditioned medium and flash-frozen mesencephalic membrane fragments. The surface markers of the differentiated neurons, such as NSE, nestin, MAP-2a, b and TH were detected by immunocytochemistry and Western blot after MSCs were cultured in induction medium for 7 days and 15 days.

RESULTSMSCs differentiated into neural progenitors and expressed nestin after MSCs were incubated with medium containing IBMX for 2 d. After the medium was replaced with induction medium containing many inducing agents, MSCs differentiated into neuron-like cells and dopaminergic neuron-like cells and expressed NSE, MAP-2a, b and TH. The percentage of NSE-positive cells, MAP-2a, b-positive cells and TH-positive cells was 30.032 +/- 2.489%, 41.580 +/- 5.101% and 34.958 +/- 5.534%, respectively after MSCs were induced in medium containing GDNF, IL-1beta, mesencephalic glial-cell-conditioned medium and flash-frozen mesencephalic membrane fragments for 15 days.

CONCLUSIONMSCs can differentiate into dopaminergic neuron-like cells and are a new cell source for the treatment of neurodegeneration diseases and have a great potential for wide application.

Adipocytes ; cytology ; Animals ; Blotting, Western ; Bone Marrow Cells ; Carboxylesterase ; analysis ; Cell Differentiation ; Cells, Cultured ; Chondrocytes ; cytology ; Culture Media, Conditioned ; Dopamine ; analysis ; Intermediate Filament Proteins ; analysis ; Mesencephalon ; cytology ; Mesenchymal Stromal Cells ; cytology ; Nerve Tissue Proteins ; analysis ; Nestin ; Neurons ; cytology ; metabolism ; Phosphoprotein Phosphatases ; analysis ; Rats ; Rats, Wistar

Objective To analyze the prognosis of pediatric burns and its influencing factors. Methods Clinical data of 1 737 children with burns from January 2013 to December 2017 in the First Affiliated Hospital of Anhui Medical University was analyzed by retrospective method. The demographic, clinical features, and related factors affecting prognosis . Results Log-binominal regression model showed that the care rate was higher in children aged 1- and 3- compared with children aged 7-12 (all P<0.05); Boiling water burns had a higher care rate than electric shock and flame burns (including chemical burn) (all P<0.05); Moderate and severe burns had a higher care rate than heavy severe burns (all P<0.05); The unhealed rate of pediatric burns in summer was higher than burned in winter (RR=0.861,95% CI:0.690-1.074); Children without complications had a higher care rate (P<0.05); Children lived in rural areas have a higher unhealed rate than lived in urban areas (RR=0.713，95% CI:0.618-0.824). Conclusions The care rate of pediatric burns was 51.1%. Major influencing factors included children aged 7-12, burned by electric and flame (including chemical burns), burned severe extraordinarily, burned in summer, and with complications, lived in rural.

Objective To study the expressions of Piwil2 protein and mRNA in papillary thyroid carcinoma(PTC) and the relationship between Piwil2 and the invasion and metastasis of PTC. Methods Immunohistochemistry and in situ hybridization were used to detect the expression of Piwil2 protein and mRNA in 60 cases of PTC with the matched adjacent non-cancerous epithelium (NCE). Results The positive rates of Piwil2 protein expression in PTC and NCE were 88.3% (53/60) and 10.0% (6/60)respectively, with significant difference ( x2 = 73. 654,P ＜ 0.01 ). The positive rates of Piwil2 mRNA expression in PTC and NCE were 85.0% (51/60) and 6. 7% (4/60) respectively, also with significant difference (x2 =74. 148 ,P ＜0.01 ). Up-regulated expressions of Piwil2 protein and mRNA were related to the invasion and metastasis of PTC (P ＜ 0.05 ). Conclusion Piwil2 may play a role in the invasion and metastasis of PTC.

Objective To study the relationship between status of methylation of human runt-related transcription factor 3 ( RUNX3 ) gene promoter in papillary thyroid carcinoma ( PTC). Methods Methylation-specific PCR and immunohistochemical SP technique were used to detect the methylation of RUNX3 gene promoter and expression of its protein in 56 cases of PTC and their matched adjacent non-carceious epithelium(NCE). Results In NCE, there was no methylation of RUNX3 gene promoter, while in PTC the methylation rate was 35.1% (20/56), which was related to the tumor TNM stage, pathological grade and lymph node metastasis (P<0. 05). The positive rates of RUNX3 protein expression in NCE and PTC were 100. 0% and 60. 7% , respectively, with a significant difference ( χ2 = 27. 378, P < 0. 05). In PTC, the positive rates of RUNX3 protein expression in grade I and grade II were 70.0% and 37.5% , respectively ( P < 0. 05 ); the rates were 46.7% and 76.9% in lymph node metastasis group and no metastasis group, respectively ( P < 0. 05 ). Moreover, there was a distinct correlation between methylation of RUNX3 gene promoter and expression of its protein (χ 2= 21. 62, P < 0. 01). Conclusions Methylation of promoter might be one of the important factors of inactivation of RUNX3 gene, and might play an important role in carcinogenesis and progression of PTC.