Objective To observe the phosphorylation level and nuclear translocation of signal transducer and activator of transcription factor-3 (STAT3) in hippocampal neurons induced by oxygen and glucose deprivation in vitro and discuss the dynamic changes of STAT3 signal pathway in an in vitro cell model of brain hypoxia and ischemia.Methods Hippocampal neurons from newly born SD rats (within 24 hours from birth) were cultured with DMEM/F12 for nine days,and then were transferred to oxygen and glucose deprivation environment for four hours to establish experimental cell models.The distribution of phosphorylated STAT3 (p-STAT3) in the hippocampal neurons in different groups was observed under laser scanning confocal microscope after immunofluorescence staining.Expression intensity of p-STAT3 at different time points after oxygen and glucose deprivation in the hippocampal neurons was detected by Western blotting.Results Expression of p-STAT3 was unobvious in the nucleus of the control group,but it was observed in the nucleus of the model group one hour after modeling,and peaked at three hour.Expression levels of p-STAT3 in the hippocampal neurons at each time point between the two groups showed significant difference (P ＜ 0.05).Conclusion Oxygen and glucose deprivation induces noticeable up-regulation of p-STAT3 in the hippocampal neuronal nucleus,which indicates the overactivation of signal transduction pathway of STAT3.

BACKGROUND:Bryan cervical disc replacement and anterior cervical discectomy and fusion have a dispute in the treatment of cervical spondylosis.
OBJECTIVE:To evaluate the clinical effects of Bryan cervical disc replacement and anterior cervical discectomy and fusion by meta-analysis, thereby providing clinical evidence for treatment strategy of cervical spondylosis.
METHODS:The authors searched Medline, PubMed, EMBASE, OVID, CBM, CNKI and also searched manual y seven relevant Chinese orthopedic journals for articles pertinent to clinical research of Bryan cervical disc replacement and anterior cervical discectomy and fusion. Extracted data included the range of motion of the cervical spine, visual analog scale score, neck disability index, the Japanese Orthopaedic Association score. Meta-analysis and forest plots were conducted with RevMan4.2.2 Software.
RESULTS AND CONCLUSION:There are eight articles in the meta-analysis, including 883 patients (430 patients receiving Bryan cervical disc replacement, and 453 patients receiving anterior cervical discectomy and fusion). Meta-analysis did detect statistical y significant differences in the range of motion of the cervical spine at 3 months and 24 months postoperatively between the two groups, but did not detect statistical y significant differences in visual analog scale score, neck disability index, the Japanese Orthopaedic Association score between the two groups. These findings indicate that Bryan cervical disc replacement is superior to anterior cervical discectomy and fusion in the range of motion of the cervical spine. However, the current literature offers no evidence to support superiority of the Bryan cervical disc replacement over the anterior cervical discectomy and fusion.

Objective To study the construction of the lentiviral-mediated RNA interference(RNAi) vector targering rat suppressors of cytokine signaling 3(SOCS3) gene.Methods Three target sequences were selected by on-line designer software on Ambion according to rat SOCS3 mRNA sequence(NM053565),the complementary DNA contained both sense and antisense oligonucleotides were designed and synthesized.After annealing,these double strands DNA were cloned to pRNA-Lenti-green fluorescent protein(GFP),which contained U6 promoter and GFP.The resulting Lentiviral vector containing SOCS3 shRNA was named pRNA-Lenti-SOCS3-GFP.After the rat glioma cells(C6)were transduced with the constructed 1entiviral vectors,real-time polymerase chain reaction was used to evaluate the level of SOCS3 expression(including siRNA1 group,siRNA2 group,siRNA3 group,vacuity group and siRNA-Negative group).The pRNA-Lenti-SOCS3-GFP and Lentivector Pakaging plasmid mix were cotransfected into 293T to package Lentivirus particles.Culture supematant was harvested,then the virus titer was determined by serial dilution assay.Results The SOCS3 mRNA sequence was successfully cloned to pRNA-Lenti-GFP,which was proved by PCR and DNA sequence.Compared with control group,the SOCS3 mRNA expressions were obviously suppressed in all 3 experimental groups,especially the expression rate in siRNA1 group was reduced by 80%.The Lentiviral particle titer was determined by serial dilution assay with 1.0?1010 TU?L-1.Conclusion The lentiviral-mediated RNAi vector of rat SOCS3 gene has been constructed successfully,this may provide a potential tool for studying and treating SOCS3-related diseases.

Indoleamine 2,3-dioxygenase (IDO) is a kind of immune regulation enzyme.IDO is the only rate-limiting enzyme that can catalyze the oxidative clearage of indole ring in tryptophan and its catabolism along the kynurenine pathway.IDO1 is one of the important mechanisms in maternal immune tolerance,which participates in the regulation of maternal-fetal immune relationship.The newly discovered IDO2 also plays an important role in this relationship.The abnormal expression of IDO2 is associated with the male infertility.

Objective To observe the phosphorylation of extraceUular signal-regulated kinase (p-ERK1/2) and its nuclear translocation at different time points after the hippocampal neurons were cul-tured in the magnesium-free medium, and discuss the changes of ERK1/2 signal pathway after epileptic injury of hippocampal neurons. Methods Hippocampal neurons from newly-born Wistar rats were cul-tured with NB medium and B-27 for 9 days, and then were transferred to the magnesium-free medium to induce epileptic injury to the hippocampal neurons. The distribution of p-ERK1/2 in the hippocampal neurons before and after the epileptic injury was observed under laser scanning confocal microscope, and the expression of p-ERK1/2 at different time points after culturing the hippocampal neurons in the magne-sium-free medium was detected by Western blot. Results Before the epileptic injury of hippocampal neurons, p-ERK1/2 mainly expressed in the cytoplasm and axoplasm of the neurons. While after the epi-leptic injury, the expression of p-ERK1/2 was detected in the cytoplasm, axoplasm and nucleus of the neurons. The expression of p-ERK1/2 was increased one hour after the epileptic injury, and peaked at hour 3 (p-ERK1:2.2838±0.1 186; p-ERK2:4.1 273±0.0 927). There was significant difference in the expression of p-ERK1/2 between the hippocampal neurons cultured with or without magnesium-free medium (P < 0.05). Conclusion Epileptic injury may induce increased expression of p-ERK1/2 in hippocampal neurons, and the activated ERK1/2 signal pathway may be associated with the epileptic dis-charge in neurons.

Objective To discuss the clinical application of noninvasive positive pressure ventila-tion for patients with acute respiratory distress syndrome (ARDS) as a result of cytomegalovirus (CMV) interstitial pneumonia after renaltransplantation. Methods There were 371 renal transplan-tation from March 2003 to October 2006, 27 patients were diagnosed as CMV pneumonia postopera-tion. Ten patients were treated with noninvasive positive pressure ventilation within the 11 patients who aggravated to ARDS. The clinical data of before and after mechanical ventilation were reviewed. Results Among patients received noninvasive positive pressure ventilation, 1 died of complication. Seven patients were cured by noninvasive positive pressure ventilation. Significant difference of the physiological index presented between the 7 patients cured with noninvasive positive pressure ventila-tion before and after the use of ventilation(P<0.05), and significant difference of the renal function also existed(P<0.05). Conclusion The major value of noninvasive positive pressure ventilation is to correct the hypoxemia.

Objective To evaluate the safety and validity of an early steroid withdrawal protocol including cyclosporine (CsA) and mycophenolate mofetil (MMF) in middle aged and elderly renal transplant patients. Methods Between September 2000 and April 2008, the prospective, randomized study design was used in 80 middle aged and elderly renal transplant patients. Steroid withdrawal group (n=39) with primary cadaveric kidney transplants received a protocol consisting of CsA 4～6 mg·kg~(-1)·d~(-1) beginning at postoperative day 3, MMF 0. 75 g twice a day from the next postoperative day, and methylprednisolone (MP) 500 mg daily from day 0 to 3. Then prednisone (Pred) 20 mg daily was gradually tapered and withdrawn after postoperative day 30. Conventional steroid treatment group (control group, n=41) received a regimen consisting of CsA, MMF and MP, and Pred 20 mg daily. Pred was tapered to 5 mg daily over a period of 6 months, then maintained thereafter. Outcome parameters were patient and graft survival rates, renal function, acute rejection ( AR), arterial hypertension, hyperlipidemia or diabetes mellitus, weight gain and infection. Results The incidence of AR in the steroid withdrawal group was similar to the control group (23. 1% vs. 19. 5%, χ~2=0. 15,P>0. 05). Patient survival rates at 12, 24, 36 months were 97. 4%, 94. 8% and 88.0% in the steroid withdrawal group and were 97.6%, 97.6 and 87.8% in the control group, respectively (χ~2=0. 17, P>0. 05). And graft survival rates were 94. 9%, 88. 6% and 83. 7% in the steroid withdrawal group and were 95. 1%, 91. 5% and 79. 5% in control group, respectively (χ~2 = 0.07, P>0. 05). Conclusions In middle aged and elderly renal transplant patients, early steroid withdrawal is feasible and may not significantly increase the risk of acute rejection episodes.

OBJECTIVEThe purpose of the present study was to investigate the in vitro effects of baicalin on induction of apoptosis in human prostate cancer cell line DU145.

METHODHuman prostate cancer cell line DU145 was treated with different concentration of baicalin in vitro. The apoptosis rate was determined by FACS analysis, cell cycle distribution was detected by flow cytometry, morphological changes and protein analysis were determined by means of electron microscope techniqueand immunohistochemical techniquerespectively.

RESULT50micromol x L(-1) and 125 micromol x L(-1) of baicalin dose-dependently induced apoptosis and inhibited the proliferation of prostate cancer cell DU145 in a dose and time-dependent manner. DNA flow cytometric analysis indicated that baicalin induced a arrest in G1 phase, showing a typical apoptosis peak. Electron microscopy detected a characteristic appearance of the apoptotic cells morphology. Immunohistochemical analysis revealed that induction of apoptosis by ways of inhibition of the bcl-2, loss of the Bax, and upregulation of Fas.

CONCLUSIONThe results indicate that baicalin may induce apoptosis and inhibit proliferation of prostate cancer cells, and has direct anti-tumor effects on human prostate cancer cells.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Flavonoids ; isolation & purification ; pharmacology ; G1 Phase ; Humans ; Male ; Plants, Medicinal ; chemistry ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Scutellaria ; chemistry ; bcl-2-Associated X Protein ; fas Receptor ; metabolism

Objective:To compare the difference between 20 cases of subcutaneous panniculitis-like T-cell lymphoma (SPTL) and 19 cases of cutaneous extra-nodal nasal-type NK/T-cell lymphoma (cutaneous NK/T-cell lymphoma). Methods:The two types of lymphoma were compared in clinical pathology, immunological marker, Epstein-Barr (EB) virus infection, and T-cell receptor (TCR) gene rearrangement. Results:Differentiated diagnosis of the two types of lymphomas was not easy based on their clinical manifestations,but the cutaneous NK/T-cell lymphoma was always followed by extracutaneous dissemination and had a poor prognosis. Histopathologically, SPTL was usually limited in subcutaneous fatty tissues while the cutaneous NK/T-cell lymphoma showed diffused infiltration around the dermis and it often infiltrated into the subcutaneous fat tissues. Coagulation necrosis, angiocentric infiltration and epidermotropism were often observed in cutaneous NK/T-cell lymphoma. When compared with immunophenotypes, SPTL often expressed βF1, membrane CD3 and CD8 but did not express CD4 and CD56, while most of the cutaneous NK/T-cell lymphomas expressed CD56 and cytoplasm CD3ε and only a few cases expressed CD3 and CD8. The differences in the expression of CD56, CD3, CD8, and βF1 were significant between the two types of lymphomas(P<0.05). The positive rate of EBER1/2 was 25% (5/20) in SPTL while it was 100% in cutaneous NK/T-cell lymphoma. The difference was statistically significant (P<0.05). Monoclonal TCR-γ gene rearrangement was found in 16 out of 20 cases of SPTL (80%) but only in 4 of 18 cases in the cutaneous NK/T-cell lymphoma (22.2%). The difference was significant(P<0.05). Conclusion:The key points to distinguish the two lymphomas are (1) extracutaneous dissemination, coagulation necrosis, angiocentric infiltration and epidermotropism; (2) the expressions of CD56, CD3, CD3ε, CD8, and βF1; (3) the positivity of in situ hybridization of EB virus; (4) detection of the monoclonal TCR-γ gene rearrangement. To make an acute differentiated diagnosis of the two lymphomas, comprehensive analysis is necessary to integrate the results of clinical manifestation, histopathology, immunophenotype, infection of EB virus and gene rearrangement.

Objective To investigate the relationship between telomerase reverse transcriptase (TERT) gene expression and astrocyte activation.Methods Twenty neonatal 3-day-old male SD rats were used for culture of the astrocytes.The astrocytes were divided into Group A (activated,non-transfected astrocytes),Group B (activated,transfected astrocytes),Group C (unactivated astrocytes) and Group D (activated,empty plasmid transfected astrocytes) according to the random number table,with 5 rats per group.The cell proliferation rate in each group was detected by cell counting kit-8 (CCK-8) ;TERT expression by immunocytochemical method; expressions of TERT and glial fibrillary acidic protein (GFAP) genes by RT-PCR assay.Results Astrocytic proliferation ability in Group B lowered significantly as compared with that in Groups A,D and C (F =43.418,P ＜ 0.01).Expressions of TERT and GFAP mRNAs in Groups A and D were significantly higher than those in Group B and C,and no significant difference was found between Groups A and D.Besides,there was a linear correlation between mRNAs expressions of both genes in Groups A and D (r =0.701,0.704,P ＜ 0.01),while no significant linear correlation was observed in Groups B and C (r =0.260,P ＞ 0.05).Expressions of TERT and GFAP proteins in Groups A and D were markedly higher than those in Groups B and C and no significant difference was found between Groups A and D.Conclusion TERT genes are involved in the activation of astrocytes and exert effect on promoting the activation of astrocytes.