By using genetic engineering techniques,we expressed in E.coli fusion proteins which con-tained human endothelial cell-derived IL-8 (EDhIL-8)、MS2 protein and different length of ?- galactosidase segments,named MS2-hIL-8、lac-hIL-8, lac-T-hIL-8 respectively.The lac-T-hIL-8 has a synthesized thrombin recognition site.Because there is a natural thrombin recognition sitewithin the EDhIL-8,thrombin can hydrolyze lac-hIL-8 and MS2-hIL-8 into natural hIL-8 ex-hibited biological activity,but has no effect on lae-T-hIL-8 which contained two recognition sitesfor thrombin.These results here indicated that the recognition of thrombin dependents on notonly the amino acid sequences of the substrates,but also the conformation formed by these aminoacids.

Objective To investigate the relationship between cytochrome P450 1A2 (CYP1A2) gene polymorphism and susceptibility to chronic obstructive pulmonary disease (COPD). Methods CYP1A2 gene polymorphisms in 100 COPD cases and 100 healthy controls were tested with polymerase chain reaction- restriction fragment length polymorphism (PCR- RELP). Results Genotype frequencies of 4 SNPs in both the COPD and control groups were in accordance with Hardy-Weinberg equilibrium (P>0.05). There was significant difference between the COPD and control groups in genotype and allele frequencies of 1D and 1F (P<0.05), but not of 1C and 1E (P>0.05). Conclusion CYP1A2*1D and CYP1A2*1F polymorphisms may play an important role in the development of COPD.

Objective To explore the best time point for the ipsilateral hepatocellular apoptosis and the contralateraI hepatic hypertrophy after selective portal vein embolization(SPVE)in rabbit.Methods In a randomized study design,forty rabbits were divided into 5 groups with 8 rabbits per-group,including one as the control and the other 4 were treated with SPVE during open surgery.The rabbits were killed postoperatively,in 3,7,14,21 days respectively after the embolization.The hepatic lobes volume,the ipsilateral hepatocellular necrosis rates and apoptosis index,and liver functions were determined as well. Results In the treatment groups,the average amount of the right liver volumes decreased from 46.4 cm~3 preoperatively to 46.0,44.4,42.0,39.7 cm~3 in groups of 3,7,14,21 days postoperatively;meanwhile,the left liver volumes increased from 54.0 cm~3 preoperatively to 54.5,56.3,61.7,63.9 cm~3 respectively during 3, 7,14,21 days after the procedures.The rates of future remaining live volumes(FRLV)increased from 53.8% preoperatively to 54.2%,55.9%,59.0%,61.0% at 3,7,14,21 days postoperatively.The apoptosis indexes of hepatocells from group A to E were 8.1%,12.2%,19.4%,20.1%,14.2% respectively.Conclusions SPVE leads to atrophy of the ipsilateral hepatic lobe and hypertrophy of contralateral lobe,indicating that hepatocytes undergone apoptosis,rather than necrosis.The time point is 7 to 14 days.

OBJECTIVETo study the characteristics of cellular metabolism of mandibular condylar chondrocytes in repairing state of osteoarthrosis and investigate its role in the pathogenesis of the disease.

METHODSTemporomandibular joint osteoarthrosis model of rabbits was created by the partial resection of joint disc and confirmed with histological diagnosis. The chondrocytes were harvested from osteoarthritic condylar cartilage in the repairing state and cultured in vitro under the monolayer culture condition. The cellular expression of cartilaginous matrix protein, collagenase and growth factors between the osteoarthritic chondrocytes and the normal controls were measured with RT-PCR technique to outline the basic feature of the osteoarthritic cells.

RESULTSThe cultured cells were confirmed as chondrocytes with their ability of expression of collagen type II and Aggrecan. In the reactive repairing state of osteoarthrosis, the chondrocytes showed the imbalance of expression of ECM proteins, and increased expression of collagenase and endogenous growth factors such as IGF-1 and TGF-beta1.

CONCLUSIONSThis study found the active anabolism of the chondrocytes within the osteoarthritic condylar cartilage and the imbalance synthesis of cartilage matrix. These repairing attempts by the osteoarthritic chondrocytes may be impossible to restore the primary homeostasis within the condylar cartilage.

Animals ; Cartilage, Articular ; metabolism ; pathology ; Cells, Cultured ; Chondrocytes ; metabolism ; Extracellular Matrix ; genetics ; metabolism ; Male ; Mandibular Condyle ; metabolism ; pathology ; Osteoarthritis ; metabolism ; pathology ; RNA, Messenger ; genetics ; metabolism ; Rabbits ; Temporomandibular Joint Disc ; pathology ; Temporomandibular Joint Disorders ; metabolism ; pathology

Aged ; Anti-Inflammatory Agents ; therapeutic use ; Cyclophosphamide ; therapeutic use ; Female ; Humans ; Immunosuppressive Agents ; therapeutic use ; Methylprednisolone ; therapeutic use ; POEMS Syndrome ; complications ; diagnosis ; drug therapy ; pathology ; Plasma Cells ; pathology ; gamma-Globulins ; therapeutic use

This study was aimed to investigate the sensitizing effect of recombinant human PDCD5 (rhPDCD5) protein on chemotherapy of U937 cell line and its mechanism. The flow cytometry was performed to assess the changes of cell apoptosis and cell cycle influenced by rhPDCD5. Hochst 33258 staining was used to observe morphology of the apoptotic cells. The activity change of caspase-3 was detected to analyse the possible mechanisms of rhPDCD5-induced apoptosis. RT-PCR was performed to observe the expression level of drug-resistant genes. The results showed that the percentage of apoptotic cells and the activity of caspase-3 remarkably increased in U937 cells treated with rhPDCD5 combined with chemotherapeutic drug; the cell cycle arrest induced by anti-tumor drug was also enhanced when combined with rhPDCD5; meanwhile, the expression levels of drug-resistant genes were down-regulated in jointly treated U937 cells. It is concluded that the chemosensitizing mechanisms of rhPDCD5 are complex. rhPDCD5 may increase the cytotoxicity of anti-tumor drugs by promoting the caspase-3-related apoptosis, influencing cell cycle, decreasing the expression of drug-resistant genes and reversing drug-resistance.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; pharmacology ; Caspase 3 ; metabolism ; Cell Cycle ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Humans ; Neoplasm Proteins ; pharmacology ; Recombinant Proteins ; pharmacology ; U937 Cells

OBJECTIVESTo evaluate the relationship between Modic change and disc height together with lumbar hyperosteogeny and study the role of Modic change in lumbar degeneration.

METHODSThe imaging data of 150 elderly patients with chronic low back pain were analysed retrospectively. All patients underwent MRI and lumbar lateral X-ray examination. The lumbar disc from L1-L2 to L5-S1 were selected for this study, including 750 discs, vertebral and endplate close to disc in 150 patients. The incidence rate of lumbar endplate Modic change, disc height and the degree of vertebral bone hyperplasia were recorded. The ratio of disc height/lumbar intervertebral disc height < 50% was defined as disc collapse. The patients were divided into 4 groups in the basis of imaging changes. Group A1:disc collapse without severe lumbar hyperosteogeny; Group A2: disc collapse with severe lumbar hyperosteogeny; Group B1: Neither disc collapse nor severe lumbar hyperosteogeny; Group B2: severe lumbar hyperosteogeny without disc collapse. The incidence rates of Modic change were compared between the 4 groups by χ(2) test. Finally, the influence of disc height and vertebral bone hyperplasia on the incidence rate of Modic change was analysed.

RESULTSFour groups of patients observed a total of 750 discs. The number of intervertebral discs in the group A1 was 208, the incidence rate was 54.3%. The number of intervertebral discs in the group A2 was 135, the incidence rate of group A2 was 34.8%. The number of intervertebral discs in the B1 group was 225, the incidence rate of group B1 was 16.9%. The number of intervertebral discs in the B2 group was 182, the incidence rate of group B2 was 29.7%. There was significant difference of lumbar endplate Modic change incidence rate among the 4 groups(χ(2) = 69.565, P < 0.05). The results of post hoc test showed that the incidence rate of Modic change in group A1 was higher than group A2, B1 and B2 (χ(2) = 12.524, 66.701 and 24.102, P < 0.00714). There was significant difference of Modic change incidence rate between group A2 and B1(χ(2) = 15.032, P < 0.00714), but there was no significant difference of Modic change incidence rate between group A2 and B2 (χ(2) = 0.945, P > 0.00714) . There was significant difference of Modic change incidence rate between group B2 and group B1 (χ(2) = 9.395, P < 0.00714).

CONCLUSIONSThe incidence rate of Modic change with disc collapse but without severe lumbar hyperosteogeny is high in elderly patients with chronic low back pain. There is no significant difference of Modic change incidence between patients with both disc collapse and severe lumbar hyperosteogeny and patients with severe lumbar hyperosteogeny but without disc collapse.

Aged ; Aged, 80 and over ; Female ; Humans ; Intervertebral Disc ; pathology ; Intervertebral Disc Degeneration ; pathology ; Low Back Pain ; pathology ; Lumbar Vertebrae ; pathology ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Retrospective Studies

OBJECTIVETo establish a multiplex RT-PCR-based reverse dot blot hybridization technique to detect influenza viruses.

METHODSObtain the HA nucleotide sequences of seasonal influenza H1N1, seasonal influenza H3N2, influenza H1N1 and human avian influenza H5N1 from GenBank. Design primers in conservative district and probes t in high variable region respectively, after analyzing the HA nucleotide sequences of influenza virus through the Vector NTI 9.0. Establish and optimize multiple RT-PCR system by comparing amplification efficiency and specificity at different primer concentrations. Establish the reverse dot hybridization system after optimizing the concentration of probes. To compare the sensitivity and specificity of this technique and the general RT-PCR Method through extracting the viral RNA of the mentioned influenza virus which are to be the reference substance.

RESULTSSuccessfully establish a multiplex RT-PCR-based reverse dot blot hybridization technique for detecting influenza viruses. This technique is 100-1000 times more sensitive than gel electrophoresis method, and it has a good specificity.

CONCLUSIONSuccessfully established multiplex RT-PCR-based reverse dot blot hybridization technique for detecting influenza viruses.

Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; isolation & purification ; Influenza A Virus, H3N2 Subtype ; genetics ; isolation & purification ; Influenza A Virus, H5N1 Subtype ; genetics ; isolation & purification ; Influenza, Human ; diagnosis ; virology ; Nucleic Acid Hybridization ; methods ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

OBJECTIVETo study the etiology and clinical significance of axial symptoms after posterior operative procedures for ossification of the posterior longitudinal ligament (OPLL).

METHODSFrom February 2005 to February 2010, 76 patients with OPLL treated were retrospectively experienced. There were 34 male and 42 female with average of 52.1 years (range from 37 to 74 years), the average duration of the disease was 32.1 months (range from 11 to 56 months). Nineteen patients underwent traditional laminectomy in group A, 33 patients received open-door laminoplasty in group B and 24 patients underwent lateral mass screw fixation in group C. All patients underwent X-ray examination pre- and post operative, computed tomography were used for diagnosis of OPLL, the recovery rate was calculated using pre- and postoperative Japanese Orthopedic Association (JOA) scores for each patient. Pre- and postoperative cervical curvature index and axial symptoms were measured and compared. χ(2) test and SNK test were used as statistical methods.

RESULTSAll patients were followed up for 14 - 35 months, average (21 ± 5) months. Loss of cervical curvature index was 4.2% ± 1.7% in group A, 2.9% ± 2.2% in group B and 2.3% ± 1.9% in group C. The difference was significant in loss of cervical curvature indice between group A and B (q = 2.94, P < 0.01), group A and C (q = 4.23, P < 0.01). The average JOA recovery rate was 58.3% for group A, 64.3% for group B and 66.7% for group C. There was no significant difference in JOA recovery rate among the three groups (P > 0.05). The rate of early evident axial symptoms was 7/19 in group A, 30.3% in group B and 33.3% in group C and the difference was not statistically significant (P > 0.05). The incidence of late evident axial symptoms was 5/19 in group A, 12.1% in group B and 8.3% in group C, the difference was not significant between group B and C (χ(2) = 13.762, P < 0.01), but of statistical difference between group A and B(χ(2) = 6.368, P < 0.01), group A and C (χ(2) = 11.481, P < 0.01). No kyphotic deformity in the group A, no "Close Door" phenomenon in group B and no internal failure in group C.

CONCLUSIONThe incidence of early axial symptoms are of no significant difference among the three groups, but late axial symptoms are higher in the laminectomy than other groups, which may be associated with loss of cervical lordosis.

Adult ; Decompression, Surgical ; adverse effects ; methods ; Female ; Follow-Up Studies ; Humans ; Laminectomy ; adverse effects ; methods ; Male ; Middle Aged ; Ossification of Posterior Longitudinal Ligament ; surgery ; Postoperative Complications ; Retrospective Studies

OBJECTIVETo explore the impact and related mechanisms of stromal cell-derived factor-1α (SDF-1α) on serum deprivation-induced apoptosis of cardiac stem cells (CSCs).

METHODSCSCs were isolated from adult mouse heart tissue and cultured in vitro. Obtained cells were purified using magnetic-activated cell sorting (MACS) with c-kit magnetic beads. C-kit(+)CSCs were divided into five groups: normal control group, serum deprivation group, serum deprivation+SDF-1α group, serum deprivation+SDF-1α+AMD3100 group, serum deprivation+SDF-1α+LY294002 group. Cell apoptosis was assessed using the DeadEnd Colorimetric TUNEL System and flow cytometry analyses with an Annexin V-FITC Apoptosis Detection Kit. The viability of CSCs was assessed by CCK-8. The protein expression of Bcl-2 and phosphorylated Akt were detected by Western blot. The caspase-3 activity was determined using caspase-3 Colorimetric Assay Kit.

RESULTSAfter magnetic separation, more than 85% of cardiosphere derived cells were positive for c-kit expression. Compared with the normal control group, the apoptosis rate of serum deprivation group was significantly increased[(27.03 ± 0.80)% vs. (1.51 ± 0.54)%, P < 0.01], which could be significantly reduced by SDF-1α in a concentration dependent manner and peak effect was seen with 100 ng/ml SDF-1α[(10.67 ± 1.06)% vs. (27.03 ± 0.80)%, P < 0.01]. The expressions of p-Akt and Bcl-2 were significantly increased and the activity of caspase-3 was significantly decreased in serum deprivation+SDF-1α group compared to serum deprivation group (P < 0.01). Further more, the expression of p-Akt and Bcl-2 were significantly decreased and the activity of caspase-3 was increased in both serum deprivation+SDF-1α+AMD3100 group and serum deprivation+SDF-1α+LY294002 group compared to serum deprivation+SDF-1α group (P < 0.01).

CONCLUSIONSSDF-1α reduces serum deprivation induced CSCs apoptosis via modulating PI3K/Akt signaling pathway.

Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cells, Cultured ; Chemokine CXCL12 ; pharmacology ; Culture Media ; chemistry ; Mice ; Myocardium ; cytology ; Proto-Oncogene Proteins c-akt ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Signal Transduction ; Stem Cells ; drug effects