Objective To explore a new peptide-based approach independent of HLA to generate antigen-specific CD+ CD8+T cells. Methods Peripheral blood mononuclear cells(PBMC) were stimula- ted for 6 h with IE-1 peptide pool. Then the activated IFN-γsecreting ceils were tested by immunomagnetic selection. And the selected cells were cultured with radio-inactivated PBMC in medium with 100 IU/ml IL-2 for 4 weeks. Results The generated T cell lines consisted of IE-1 specific CD4+ T (6.88%) and CD8+ T cells 92.99%, which demonstrated antigen-specific killing and cytokine secretion. Conclusion T ceils can be proliferated with this new procedure, and maintain its phenotype and antigen-specific function.

OBJECTIVE To discuss risk factors of hospital infection in patients who suffered from primary cancer of the liver in the medium and advanced stage and give the countermeasures.METHODS The liver cancer cases in medium and advanced stage in 2001-2006 were retrospectively investigated and analyzed.RESULTS The hospital infection rate was 33.92% for the liver cancer patients who were in the medium and advanced period;the more common sites of infection were lungs,abdominal cavity,intestinal tract,blood.The motality was 20.83% in the infection group and 10.16% in non-infection group and there were significant differences between two groups(P

Objective To investigate the efficacy of sequential add-on of pegylated interferon α-2a (PEGIFN-α-2a) for 48 weeks in chronic hepatitis B (CHB) patients with low serum hepatitis B e antigen (HBeAg) titer after long term adefovir dipivoxil (ADV) monotherapy.Methods This was a randomized,open and prospective clinical trial.Patients who had been treated with ADV for 72 to 144 weeks,with undetectable serum hepatitis B virus (HBV) DNA level,low HBeAg titer (5 S/CO＜ HBeAg＜50 S/CO) and serum hepatitis B surface antigen (HBsAg) ＜5000 IU/mL were included.The patients were categorized into ADV monotherapy group and ADV plus PEGIFN-a-2a combination therapy group by random number table.Patients in ADV group continued ADV monotherapy and patients in combination therapy group added PEGIFN-α-2a to ADV for 48 weeks.After the treatment,efficacy of the two therapies were assessed by comparing the reduction of serum HBeAg reduction,HBeAg loss rate,HBeAg seroconversion rate,and reduction of intrahepatic HBV DNA and HBV covalently closed circular DNA (cccDNA).Pre-and post-treatment results were compared by paired samples t test.Comparison between groups was performed using indepedent samples t test.Comparison of rates between groups was performed using x2 test.Results The trial enrolled 55 CHB patients,and there were 27 patients in ADV monotherapy group,28 patients in combination therapy group.Baseline characteristics including age distribution,sex ratio,alanine aminotransferase (ALT),aspartate aminotransferase (AST),total bilirubin (TBil),serum HBeAg and HBsAg,hepatic HBV DNA and HBV cccDNA were all comparable (all P＞0.05).Twenty-five patients in ADV monotherapy group and 26 patients in combination therapy group completed 48 weeks treatment.HBeAg loss rates and seroconversion rates of combination therapy group were higher than those of ADV monotherapy group (x2 =5.38 and 4.69,respectively,both P＜0.05).HBeAg titers of both groups were significantly lower than those of baseline (t=8.43 and 8.50,respectively,both P＜0.05).The HBeAg titer of combination therapy group was lower than that of monotherapy group (t=5.60,P＜ 0.01).HBV DNA and HBV cccDNA in liver tissue of combination therapy group was (6.934±0.52) lg IU/mg and (5.63±0.54) lg IU/mg post-treatment,respectively,which were both lower than baseline (t＝7.12.6.67,respectively,both P＜0.01).HBV DNA in liver tissue of monotherapy group was (7.09＝0.43) lg IU/mg post-treatment,which was lower than baseline (t=2.67,P=0.02).After treatment,HBV cccDNA in liver tissue of combination therapy group was lower than that of monotherapy group (t =2.87,P=0.00).Conclusions Compared with ADV monotherapy,sequential add-on of PEGIFN-a-2a in combination with ADV can achieve higher serum HBeAg loss rate and seroconversion rate and facilitate the clearance of hepatic HBV DNA and HBV cccDNA in CHB patients with low HBeAg titer after long-term ADV monotherapy.

Objective To investigate the clinical characteristics of postoperative spondylodiscitis after lumbar intervertebral disc herniation.Methods 9 cases with spondylodiscitis were reviewed,about their clinical symptoms,laboratory tests,bacteria culture,CT and MRI finging,etc.Results and Conclusion The clinical symptoms occurred 8~24 d after operation.Laboratory test and MRI were helpful for the early diagnosis.The outcome of both surgical and non-surgical treatment was satisfactory.

Objective:To study the effect of OSAHS on adult nasal-cycle.Method:The nasal cycle of 20 healthy adults,18 patients of snoring and 22 patients of OSAHS were examined with acoustic rhinometry,which was performed every 30 minutes in 7 hours.Result:The ratio of nasal cycle in healthy adults was(19/20,95.0%), in snoring patients was(15/18,83.0%),in patients of OSAHS was(15/22,68.2%). The mean alteration amplitude of nasal cycle in healthy adults was significantly larger than that in patients with OSAHS (P<0.05).The distribution of the healthy adults and patients with OSAHS between the typical cycle categories was significantly different(P<0.05).Conclusion:The characteristics of nasal-cycle in of OSAHS patients were different with healthy adults,which maybe owing to change of physical function of nasal cavity.

Objective To investigate the effect of bone marrow mesenchymal stem cells (BMSCs) transplantation on angiogenesis and functional recovery after focal cerebral ischemia in rats.Methods BMSCs were isolated and cultured using the whole bone marrow adherent method,and conducted phenotypic identification using flow cytometry analysis of surface positive antigen of CD29,CD90 and the negative antigen of CD34,CD45.Rats were subjected to middle cerebral artery occlusion (MCAO) for 90 minutes,and divided into three groups randomly,the sham group,model group and BMSCs group.24 hours after cerebral ischemia,rats were injected with 1 ml BMSCs solution (1 × 106 cells/ml) or PBS via the tail vein.The modified neurological severity score(mNSS) test,the corner test and the adhesive tape test were used to evaluate sensorimotor function on the 1,7,14 and 28 days after ischemia.Infarcted volume was detected by toluidine blue staining,and the numbers of vWF positive microvessels and vascular endothelial growth factor (VEGF) positive cells in the ischemic boundary were determined by immunofluorescence.Results By flow cytometric analysis,the cell phenotype of passage 3 BMSCs showed that CD29,CD90,CD34 and CD45 were 98.3％,97.4％,0.2％ and 4.8％,respectively.Compared with the model group,BMSCs significantly reduced the score of mNSS(P＜0.01),the number of right turn of corner test(P＜0.05),latency of removal adhesive tape(P＜0.05) and the infarcted volume (P＜0.01).The numbers of vWF positive vesscls and the VEGF positive cells were (42.97±8.64)/mm2 and (54.83± 10.66)/mm2 at the boundary zone in model group 14 days after ischemia,respectively.BMSCs significantly increased the numbers of vWF positive vessels ((69.43± 7.29)/mm2) and VEGF positive cells ((78.70±6.16)/mm2,P＜0.01).Conclusion BMSCs can improve the functions of cerebral lesions after cerebral ischemia,which may be associated with the enhanced angiogenesis and VEGF expression in the ischemic boundary.

BACKGROUND:Studies have shown that umbilical cord blood stem cel transplantation promote the recovery of spinal cord injury, and electroacupuncture also can inhibit the proliferation of astrocytes to reduce damage to scar formation, suggesting that a combination of umbilical cord blood stem cel transplantation and electroacupuncture may play an important role in the treatment of acute spinal cord injuries. OBJECTIVE:To observe the influence of transplantation of human umbilical cord blood stem cels combined with electroacupuncture at theDu channel on expression of nerve growth factor and neurotrophin 3 in rats with spinal cord injuries. METHODS: Seventy-two female Sprague-Dawlay rats were randomly divided into control group, injury group, transplantation group and combined therapy group. In the control group, only an incision on the back was sutured;in the injury group, a piece of saline-infiltrated gelatin sponge, 1 mm×2 mm×2 mm, was placed into the transected spinal cord at T10 level; in the transplantation group and combined therapy group, a piece of gelatin sponged infiltrated in the suspension of human umbilical cord blood stem cels was placed into the transected spinal cord, respectively, and then, electroacupuncture stimulation at the Duchannel was performed in the combined therapy group at 1 hour after modeling. Specimens were taken at 7, 14, 28 days after modeling in each group, and then immunohistochemistry, western blot and real time-PCR methods were used to detect the expression of nerve growth factor and neurotrophin 3. RESULTS AND CONCLUSION:Compared with the transplantation group, the expression of nerve growth factor and neurotrophin 3 was lower in the injury group but higher in the combined therapy group at 7, 14, 28 days after modeling (P < 0.05). The results of western blot and real time-PCR were consistent with those of immunohistochemical detection. Findings show that human umbilical cord blood stem cel transplantation combined with electroacupuncture has a remarkable synergistic effect in the treatment of spinal cord injury that can significantly up-regulate the expression of nerve growth factor and neurotrophin 3, and contribute to injured spinal cord repair, regeneration and functional recovery after spinal cord injury.

AIM:To investigate whether Smad pathway participates the process of extracellular signal regulated kinase (ERK) induced the proliferation of vascular smooth muscle cells (VSMCs). METHODS: Human umbilical artery smooth muscle cells (hUASMCs) were divided into four groups: control group, PDGF (platelet derived growth factor) group, ERK blocking agent group and PDGF+ERK blocking agent group. MTT assay was used to detect the proliferation of hUASMCs (A value). Immunohistochemical technique was used to detect the expression of PCNA, phosphorylated ERK (p-ERK) and phosphorylated Smad2/3 (p-Smad2/3) protein in hUASMCs. The expression of Smad2/3 mRNA in hUASMCs was detected by RT-PCR. RESULTS: The proliferation of hUASMCs and the expression of PCNA, p-ERK and p-Smad2/3 proteins in hUASMCs in PDGF group were increased obviously than those in other groups (P

ObjectiveTo explore the relationship between chronic kidney disease (CKD) and cerebral small vessel disease (SVD) in elderly patients. MethodsOne hundred and fifty-two elderly male CKD patients for experimental group and 158 elderly male for control group were recruited. Demographic data and vascular risk factors were recorded. White matter lesion (WML) was semi-quantitatively assessed by cerebral magnetic resonance imaging (MRI), and lacunar infarction (LI) was also calculated. Results(1) The prevalenees of hypertentsion and diabetes mellitus were higher in elderly CKD patients than those in control group (30. 9% vs. 19.0%, 23.7%vs. 14.6%;both P～0. 05). (2) The percentages of grade 2 and grade 3 WMLs were higher in elderly CKD patients than those in control group (34.9% vs. 24.1%, 25.7% vs. 16.5%;both P<0.05). Prevalence of LI was higher in elderly CKD patients than that in control group (45.4% vs.25.3% ,X2= 13. 70, P<0. 05). The similar Resultswere also obtained except for control subjects with hypertension and diabetes mellitus. (3) The logistic regression analysis showed that age, hypertension and low glomerular filtration rate (GFR) were closely associated with SVD in elderly CKD patients. ConclusionsHypertention and diabetes mellitus are risk factors for CKD in elderly patients. SVD is associated with CKD, and age, hypertension and low GFR may be risk factors for SVD in elderly CKD patients.

Objective To explore the strategy and outcomes of surgical treatment of thoracic ossification of ligamentum flavum(OLF),especially combined with ossification of posterior longitudinal ligament,thoracic kyphosis and epidural adhesion.Methods Fifty-three cases of thoracic OLF from January 2003 to December 2009 were reviewed retrospectively.All patients were treated by the methods of en bloc resection of semi-facet and lamina.All patients were followed up for more than half an year,including 32 males and 21 females,aged from 43 to 73 years(average 54.7 years).The lesions located in upper thoracic for 18 patients,and in thoracolumbar for 35 patients.For multi-level or jumping OLF patients,the responsible levels were determined by combination of images and clinical symptoms.For multi-level OLF with ossification of posterior longitudinal ligament(OPLL)or thoracic kyphosis(＞50°),multi-level pedicle screw fixation and correction of kyphosis were performed.For dural adhesion patients,part of cerebrospinal fluid was released with a caudal incision of dural sac resulting in collapse and epidural arachnoid separation.Ossific and adhesion dura mater were removed with integrity of arachnoid.The surgical outcomes were evaluated with preoperative and postoperative thoracic Japanese Orthopaedic Association(JOA)score,Nurick grade and neurologic functional recovery ratio.Results Fifty-three cases were followed up for 6 months to 6 years,with an average of 18 months.The average preoperative JOA score was 4.3±2.3,which significantly increased to 8.3±1.8 after operation.Postoperative neurologic functional recovery rates were 11% to 80%(average 65.8%),including excellent in 18 cases,good in 20,fair in 10,and poor in 5.The excellent or good rate was 71.7%.The mean preoperative Nurick grade was 3.7(2-5 grade)and decreased to 2.3 grade after operation.Conclusion En bloc resection of semi-facet and lamina is a safe and effective method for treatment of thoracic OLF.For the patients with OPLL or kyphosis,pedicle screws fixation and kyphosis correction was beneficial for recovery of neurologic function of thoracic OLF patients.