Objective:To investigate the most effective strategy for mature induction of dendritic cells.Methods:Human monocyte-derived dendritic cells were induced in the presence of cytokines GM-CSF and IL-4. On day 6, the immature DCs were pulsed with each of CD40L, LPS, TNF-? or a cocktail of cytokines(TNF-?, IL-6, IL-1?, PGE2). DCs were harvested after 24 h induction. The surface markers for maturation CD80,CD83,CD86 and HLA-DR were detected by FCM. FITC-Dextra endocytic activity was measured by FCM. IL-12 production was detected by ELISA. The capacity of DCs for T cell activation was detected by MTT assay.Results:CD40L,LPS,TNF-? and the cocktail of cytokines all could induce DCs’ maturation. The most effective scheme for induction of maturation was the cocktail of cytokines, and the expression rate of CD83 was up to 66.91%(P

Objective To investigate the feasibility and safety of autologous tumor tissue lysate loading den-dritic cells(DC) for the treatment of hepatocellular carcinoma (HCC). Methods The monocytes-derived DC were induced and antigen loaded with tumor tissue lysate to produce DC vaccine. Vaccination and clinical observation were conducted in 12 HCC patients for 41 times. Results The average output of DC was 1.69×107(1.69×107±9.44×106>) from 90 ml peripheral blood. 63.41% (26/41)patients appeared to develop delayed-type hypersensitivity after intradermal injection. After an average of 9 months follow up, 1 patient out of 4 recurrence and metastasis pa- tients survived for 17 months. The other three patients progressed. Out of 8 patients undergoing immunotherapy post- operatively,6 patients had no signs of recurrence and the others were found to have liver rceurrence and progression. Conclusion DC based immunotherapy is safe and feasible,with no side effects,which can be applied in the immu- notherapy strategy of HCC patients.

To reduce the mismatching and non-matching in the protein two-dimension electrophoresis (2-DE) images, we proposed an auto-matching algorithm based on gray hierarchical and geometric blocking in this study. Firstly, protein spots in the gel images were divided into groups by gray level and geometric position, and then a method based on shape context and normalized correlation was used for coarse matching in protein spots. Secondly, matched pairs in coarse matching were set as feature points, and the precise matching in the rest of not matched protein spots was accomplished by the method of geometric correlation and similarity criterion. Finally, local affine transformation was used in the verification of matching results to remove non-matching and mis-matching points. The algorithm was applied to different 2-DE gel images. The results showed that the new matching algorithm could reduce the non-matching and mis-matching spots, and increase the matching accuracy.
Algorithms ; Electrophoresis, Gel, Two-Dimensional ; Image Processing, Computer-Assisted ; Proteins ; analysis

Background As one of the most common microvascular complication of diabetes in eyes,diabetic retinopathy (DR) is one of the most important cause of blindness.Nuclear factor-kappa B (NF-κB) is involved in the occurrence and development of the disease through the activation of a series of inflammatory cytokines.Objective The present study was to investigate the effects of peroxisome proliferator-activated receptor-gamma (PPAR-γ) excitomotor,rosiglitazone,on NF-κB expression and apoptosis of the retinal ganglion cells (RGCs) in the retina with diabetes mellitus. Methods Ninety SPF male Wistar rats were randomized into normal control group,diabetic control group and rosiglitazone group.Diabetes mellitus was induced by intraperitoneal injection of 50 mg/kg streptozotocin(STZ).Then 3 mg/kg rosiglitazone was intragastricly administered once per day in the rosiglitazonegroup,and the same volume of saline solution was used at the same way in the normal control group and diabetic control group from 3 days after modeling.The rats were sacrificed and the eye cups specimen was made at 4,8 and 12 weeks after usage of drugs.Retinal histopathological examination was performed by hematine-eosin staining,and expression of NF-κB p65 protein in retina and apoptotic index(AI) of RGCs were detected by immunohistochemistry and TUNEL assay,respectively in different time points mentioned above.The use of the animals complied with the Regulations for the Administration of Affairs Concerning Experimental Animals by State and Technology Commission.Results The blood glucose level was significantly elevated at various time points in the diabetic control group and rosiglitazone group compared with normal control group (P＜0.01 ),and that of the rosiglitazone group was significantly declined in comparison to the diabetic control group (q =0.81,0.82,1.23,P＞ 0.05 ).Normal retinal structure was seen in the normal control group,and edema retinal cell and disorder of retinal layers were exhibited in the diabetic control group.Retinal structure was almost normal in the rosiglitazone group.The NF-κB p65 was expressed weakly in the retina of normal control group,but the expression of NF-κB p65 was significantly elevated in the diabetic control group and rosiglitazone group compared with the normal control group(P＜0.01 ).However,the expression of NF-κB p65(A value)was significantly decreased in the rosiglitazone group compared with diabetic control group at 8 weeks and 12 weeks( q=17.77,15.30,P＜0.01 ).There were a few apoptotic cells in rat retina of the normal control group.Compared with the normal control group,the AI of the diabetic control group and rosiglitazone group was significantly reduced(P＜0.01 ).However,the AI of RGCs in the rosiglitazone group was significantly lower than that of diabetic control group in various time points (q =19.28,27.39,49.92,P＜0.01 ). Conclusions As one of the PPAR-γexcitomotors,rosiglitazone can inhibit apoptosis of RGCs through downregulating the expression of NF-κB in rat retina with diabetes mellitus,indicating a protective effect of rosiglitazone on retina in diabetic rat.

Objective To study the expressions of Toll-like receptor 9 protein (TLR9) in peripheral B and T lymphocytes in newly diagnosed, untreated patients with systemic lupus erythematosus (SLE) and their relationship with clinical parameters. Methods Blood samples were obtained from 35 newly diag-nosed, untreated patients with SLE and 16 healthy human controls. B, T lymphocytes and TLR9 protein were labeled with fluorescent antibodies, and the expressions of TLR9 protein were detected by flow cytometry in peripheral B and T lymphocytes. The relationship between TLR9 expression and clinical parameters was assessed. Results The proportions of B and T lymphocytes expressing TLR9 in newly diagnosed, untreated patients were (53.94±17.95)% and (49.33 ± 23.30)%, respectively, compared to (29.40 ± 10.54)% and (29.18 ± 14.78)%, respectively, in healthy controls (t = 6.11,3.73, respectively, both P < 0.01). Additionally,the proportion of B lymphocytes expressing TLR9 correlated negatively with SLE disease activity index (SLEDAI)(r = -0.39, P < 0.05), but positively with the level of serum IgA antibody (r = 0.74, P < 0.01).Condnsions The expression of TLR9 is elevated in peripheral T and B lymphocytes from patients with newly diagnosed, untreated SLE, and the proportion of TLR9-expressing B lymphocytes negatively correlates with SLEDAI, but positively correlates with the serum level of IgA antibody.

Objective To observe the effects of early rehabilitation on the serum neuron specific enolase (NSE) levels of patients with acute cerebral infarction.MethodsSixty patients with acute cerebral infarction were randomly divided into a rehabilitation group and a control group. All received routine treatment at the acute stage, including anti-platelet aggregation medication, drugs for improving microcirculation, neurotrophic agents and prompt treatment of any complications. Patients in the rehabilitation group also received systemic rehabilitation training beginning immediately after their vital signs had been stabilized. NSE in serum was assayed before treatment and after 3, 7and 14 days. National Institutes of Health stroke scale (NIHSS) scores were evaluated at each time point, and the two groups were compared.ResultsThere was no significant difference in serum NSE or NIHSS scores between the two groups pre-treatment. Both groups improved to a certain extent, but the improvements in the rehabilitation group were significantly better than in the control group, as their NSE levels at 7 days and NIHSS scores at 14 days were both significantly better.ConclusionsEarly rehabilitation intervention contributes to reducing serum NSE levels after acute cerebral infarction, lessening brain injury, and thereby promoting the recovery of damaged neural function.That may be one of the mechanisms by which early rehabilitation promotes functional recovery in patients with acute cerebral infarction.

ObjectiveTo observe the effect of lappaconitine (LA) on brain water content and serum IL-I,IL-2 and TNF-α levels in rats with traumatic brain injury (TBI) as well as its cerebral protective function.MethodsA total of 24 male SD rats were involved in the study and randomly divided into control group,TBI group and TBI +LA group,with eight rats per group.The rats in the TBI group and TBI + LA group were inflicted with fluid percussion injury ( FPI ).The rats in the TBI + LA group were treated with LA (4 mg/Kg/d,ip,for 10 consecutive days).The neurological score,brain water content and serum IL-I,IL-2 and TNF -α concentrations were detected at time points including TO ( before FPI ),T1 (one day after FPI),T2 (five days after FPI) andT3 (10 days after FPI).Results At each time point after FPI,the neurological dysfunction was observed in both the TBI group and TBI + LA group.The neurological dysfunction was gradually alleviated from TI to T3 in the FPI + LA group,which showed significant lower neurological score as compared with the TBI group (P ＜0.05 or 0.01 ).The brain water content in the TBI group and TBI + LA group was significantly higher than that in the control group at each time point after FPI.Meanwhile,the water content of the TBI + LA group was significantly lower than that of the TBI group ( P ＜ 0.01 ).The serum IL-1,IL-2 and TNF-alpha concentrations in the TBI group and TBI + LA group were significantly higher than those in the control group at each time point after FPI,and the serum IL-I,IL-2 and TNF-αt concentrations of the TBI + LA group were significantly lower than those of the TBI group ( P ＜ 0.01 ).Conclusions LA exerts cerebral protective effects of TBI rats by relieving the neurological dysfunction and cerebral edema and reducing the serum IL-1,IL-2 and TNF-α concentrations.

BACKGROUND:Open fracture of lower limb with severe soft tissue and bone defects also accompanies anterior tibial soft tissue defects and exposure of sclerotin and steel plate, which can be crucial y treated with strong fixation and use of skin flap to block the wound.
OBJECTIVE:To explore the clinical efficacy of a large area of soft tissue defects in the anterior tibia using vacuum sealing drainage combined with groin free flap.
METHODS:A total of 24 patients with a large area of soft tissue defects in the anterior tibia were included in this study and then divided into two groups, with 12 cases in each group. In vacuum sealing drainage group, the scope of soft tissue defects was ranged from 10 cm×15 cm to 15 cm×20 cm. After the debridement, the fracture was fixed with external fixation scaffold and the wound was covered with the vacuum sealing drainage dressing. The blood clot was rinsed with normal saline via T-tube, and 7-10 days later the vacuum sealing drainage was given. According to the growth of granulation tissue, the wound was secondarily sutured, fol owed by groin free skin flap of superficial iliac circumflex artery with medial knee arteriovenous anastomosis transplantation. In the non-vacuum sealing drainage group, the wound size was ranged from 10 cm×5 cm to 30 cm×20 cm, the period from injury to admission was 1-24 hours. They were given conventional debridement and secondary fixation or skin flap transplantation.
RESULTS AND CONCLUSION:The length of preoperative hospital stay and the skin flap are in vacuum sealing drainage group were significantly better than those in non-vacuum sealing drainage group (P<0.05). There was no significant difference in the length of postoperative stay and total length of hospital stay between the two groups (P>0.05). The wound infection rate was 0 in vacuum sealing drainage group and 75%in non-vacuum sealing drainage group at 8-14 days after treatment. The wound and donor area incision were healed at I stage, the skin grafts survived. Al the involved patients in two groups were fol owed up, for 6-36 months. During the fol ow-up process, the fracture healing time in non-vacuum sealing drainage group was significantly longer than that in vacuum sealing drainage group. The skin flap in two groups was similar to surrounding skin in color and texture, the flap exhibited no vessels, no ulceration, and no clumsy. The vacuum sealing drainage combined with groin free flap can timely control a large area of soft tissue defects post-trauma, improve wound blood supply, shorten preoperative preparation time, early close the wound, significantly promote the healing of wound and fracture. The skin flap is soft, flexible, wel-looking, and active functional, it significantly shortens the course of treatment and maximizes the recovery of limb function.

Objective To investigate the effect of curcumin on apoptosis in hippocampal neurons and the expression of c-Jun N-terminal kinase 3 (JNK3) and postsynaptic density protein 95 (PSD95) in hippocampus during cerebral ischemia-reperfusion (I/R) in rats with spontaneous hypertension (SH) .Methods One hundred and thirty-five male rats (homologous with WKY) with SH and 90 male normotensive WKY rats, weighing 275-325 g,were used in this study. The WKY rats were randomized into 2 groups ( n = 45 each) : sham operation group (WS group) and cerebral I/R group (W-I/R group) . The rats with SH were randomly divided into 3 groups ( n = 45each) : sham operation group (S-S group), cerebral I/R group (S-I/R group) and curcumin group (S-C group) .Global cerebral ischemia was produced by 4 vessel-occlusion method. The bilateral common carotid arteries were only exposed but not ligated in W-S and S-S groups. Intraperitoneal corn oil 10 ml/kg was injected at 30 min of reperfusion in W-I/R and S-I/R groups. Intraperitoneal curcumin 100 mg/kg was injected at 30 min of reperfusion in S-C group. Three animals in each group were sacrificed at 2 h, 6 h, 1 d, 3 d and 7 d of reperfusion and their brains were harvested for determination of apoptosis in hippocampal neurons and the expression of JNK3 and PSD95in hippocampus. Results The number of apoptotic neurons was significantly increased in S-S group compared with W-S group ( P < 0.05) . The number of apoptotic neurons was significantly increased and the expression of JNK3was up-regulated in S-I/R group compared with S-S group ( P < 0.05) . The number of apoptotic neurons was significantly decreased and the expression of JNK3 was down-regulated in S-C group compared with S-I/R group (P <0.05) . There was no significant difference in the expression of PSD95 among all the groups ( P > 0.05) . Conclusion Curcumin can inhibit apoptosis in hippocampal neurons and the mechanism is related to down-regulation of the expression of JNK3 in hippocampus. The mechanism by which curcumin down-regulates the expression of JNK3in hippocampus may not be related to PSD95 pathway.

Objective: To investigate the impact and its possible mechanism of hydrogen sulfide (H2S) on myocardial collagen remodeling in experimental rats with diabetic mellitus (DM).
Methods: Rat’s DM model was established by intraperitoneal injection of STZ at 40 mg/kg. A total of 40 SD rats were randomly divided into 4 groups:Control group, DM group, DM+NaHS group, in which NaHS worked as exogenous donor of H2S and NaHS control group. n=10 in each group, all animals were treated for 8 weeks. The cardiac collagen deposition was observed by Masson staining, protein expressions of cardiac collagen types I, III, IV and transforming growth factorβ1 (TGF-β1), connective tissue growth factor (CTGF) were examined by Western blot analysis.
Results: Compared with Control group, DM group showed increased protein expressions of cardiac collagen types I and III, up-regulated expressions of TGF-β1 and CTGF, P<0.05;while the expressions of collagen type IV were similar between 2 groups. Compared with DM group, DM+NaHS group presented reduced cardiac collagen expression, decreased expression of collagen types I and III, down-regulated expressions of TGF-β1 and CTGF, P<0.05;while the expressions of collagen type IV were similar between 2 groups.
Conclusion: H2S may improve the myocardial collagen remodeling in experimental DM rats, the mechanism might be related to the down-regulation of TGF-β1 and CTGF expression.