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 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.

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 immunogenicity and immunoprotective effect of heat shock protein 60 kDa (SjHSP60) of Schistosoma japonicum in mice after immunization and challenge infection, and explore the mechanism. Methods B cell/an?tibody?related databases and analysis tools were used to predict B?cell epitopes of SjHSP60. The mice were immunized with the recombinant SjHSP60 and challenged with S. japonicum cercariae. SjHSP60?specific antibodies in serum were detected by ELI?SA. The level of splenocyte proliferation was determined by 3H?TdR incorporation. Ex vivo suppression assay was performed to in?vestigate the effects of CD4 +CD25 + regulatory T cells (Tregs) induced by SjHSP60. Results SjHSP60 possessed multiple pre?dominant regions of B?cell epitopes. SjHSP60 induced a significant increase in both SjHSP60?specific IgG levels (P < 0.01) and splenocyte proliferation (P < 0.01) with a higher IFN?γ production (P < 0.01). However, the immunization with SjHSP60 resulted no significant reduction in adult worms (P > 0.05) and liver?accumulated eggs (P > 0.05) in S. japonicum?infected mice. Ex vivo assay showed that CD4+CD25+ Tregs from SjHSP60?immunized mice enhanced immunosuppressive activity. Conclusion SjH?SP60 has a dual role in host immune system, being involved in the induction of dominant humoral and cellular immune responses as well as in the enhancement of immunosuppression.

BACKGROUND:The main treatment of lumbar isthmic spondylolisthesis is the surgery, in a broader attempt to decompression, reduction, fixation and fusion of the lesioned segments. The golden standard of the treatment is biological fusion, while internal fixation is a reliable assistance for fusion therapy. ＜br> OBJECTIVE:To discuss the clinical value and curative effect of intervertebral fusion cage combined with pedicle screw systems for the treatment of lumbar isthmic spondylolisthesis. ＜br> METHODS:From March 2010 to March 2013, 21 cases of isthmic spondylolisthesis were treated with intervertebral fusion cage combined with pedicle screw systems, including 18 cases of spondylolisthesis of degree II and 3 cases of spondylolisthesis of degree III. Al patients were fol owed up regularly, taking JOA lumbago score and visual analog scale score as the objective evaluation criteria of pain in postoperative fol ow-ups. The curative effect was assessed by Macrab standard, and the functional recovery was evaluated based on indicators such as Prolo, and the spinal fusion rate was assessed according to Lenke criteria. Changes of slippage rate, slippage angle, sacral inclination angle and intervertebral space post height in preoperative and postoperative periods were evaluated by iconography data. ＜br> RESULTS AND CONCLUSION:Al the 21 patients with isthmic spondylolisthesis were fol owed up for 12-16 months. JOA lumbago score and vasual analog scale score of al patients were improved after treatment, and the difference was statistical y significant compared with before treatment (P=0.000). According to Macrab evaluation criteria, there were 17 excellent cases and 4 good cases. Each indicator evaluated by preoperative Prolo activities and symptom grading showed significant differences in preoperative and postoperative periods (P=0.003). Postoperative lumbar spondylolisthesis was basical y reset, the slippage angle was significantly reduced, the sacral inclination angle was increased, and the height of the intervertebral space was recovered basical y. Intervertebral fusion cage combined with pedicle screw systems was one of the effective strategies to treat lumbar isthmic spondylolisthesis.

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.