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

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.

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

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 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.

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 evaluate impacts of acute hypervolemic hemodilution (AHH) and intra-operative cell salvage (ICS) with 6％ volume fraction of hydroxyethyl starch (HES) on hemodynamics,blood saving efficiency and renal function of orthopedic surgery patients.Methods A total of 58 patients from orthopedic surgery were involved and randomly divided into AHH + ICS group (30 cases) and control group (28 cases).Changes of hemodynamic indices (HR,MAP and CVP) and renal function indices (BUN,BCr,UCr and ALB) in both groups were compared before operation (T0),immediately after operation (T1) and at postoperative 4 hours (T2),1 day (T3) and 2 days (T4).CCr was counted and intraoperative blood conservation was observed at each time point as well.Results HR,MAP and CVP of the two groups had no significant differences.Both groups showed some drop of HR (P ＜ 0.05),but an increase of MAP and CVP at T1-T4 (P ＜ 0.05),in contrast with levels at TO.BUN,BCr and ALB also showed insignificant differences between groups or within group at each time point.CCr in the control group showed no significant difference at each time point.On the contrary,CCr in the AHH + ICS group had a fall at T1-T4 and was declined to the lowest level at T2.CCr in the AHH + ICS group showed a recovery at T3-T4 and its level at T4 was still lower than that at TO,with no significant difference.CCr in the two groups showed insignificant difference at TO,but its level in the AHH + ICS group was lower than that in the control group at T1-T4,at T2 in particular (P ＜0.01).Moreover,CCr in the two groups was still significantly different at T4 (P ＜ 0.05).Renal function indices of the two groups were all within normal range at each time point.Intraoperative blood loss and unrine volume of the two groups had no significant differences,but intraoperative fluid requirement,allogenic blood transfusion volume and transfusion rate of AHH + ICS group were notably lower than those of control group (P ＜ 0.05 or P ＜0.01).Conclusions AHH plus ICS using HES are safe,effective and promising integrated blood conservation measures,which significantly reduces intraoperative allogenic blood transfusion volume and transfusion rate and has few impacts on fundamental vital signs and renal function.However,prolonged use of large dose of HES may do harm to renal function and therefore should be carefully considered in treatment of patients with severe renal dysfunction.

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.

Objective To investigate the effect of curcumin on the expression of cannabinoid receptor 1 (CBR1) and NR2B subunit-containing NMDA receptor in spinal cord dorsal horn and dorsal root ganglion (DRG) neurons in a rat model of neuropathic pain.Methods Seventy-two male Sprague-Dawley rats,weighing 200-230 g,were randomly divided into 4 groups ( n =18 each):sham operation group (S group),chronic constrictive injury (CCI) group,curcumin group (Cur group) and solvent control group (SC group).Neuropathic pain was induced by CCI.The sciatic nerve was exposed and 4 loose ligatures were placed on the sciatic nerve at 1 mm intervals in groups CCI,Cur and SC.Curcumin was injected intraperitoneally at 100 mg· kg- 1· d-1 for 14 consecutive days after operation in Cur group,while the equal volume of dimethyl sulfoxide was given instead of curcumin in SC group.Mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were measured on 2 days before operation and on 1,3,7,10 and 14 days after operation.Six rats in each group were sacrificed on 3,7 and 14 days after operation and the lumbar segments of the spinal cord ( L4,5 ) and DRGs were removed to determine the expression of CBR1 and NR2B by immuno-histochemistry.Results Compared with S group,MWT was significantly decreased,TWL was significantly shortened,and the expression of CBR1 and NR2B in spinal cord dorsal horn and DRG neurons was up-regulated after operation in the other three groups (P ＜ 0.05 ).Compared with CCI group,MWT was significantly increased,TWL was significantly prolonged,the expression of CBR1 inspinal cord dorsal horn and DRG neurons was up-regulated,and the expression of NR2B in spinal cord dorsal horn and DRG neurons was down-regulated after operation in group Cur (P ＜ 0.05 ),and no significant change in the parameters mentioned above was found in group SC ( P ＞ 0.05 ).Conclusion Curcumin can alleviate neuropathic pain in rats,and up-regulation of CBR1 expression and down-regulation of NR2B expression in spinal cord dorsal horn and DRG neurons are involved in the mechanism.

Phase Ⅰ Clinical trial of new drugs is key for clinical drug development. In this article,we describe our experience and reflection on phase Ⅰ clinical trials of new drugs at the First Affiliated Hospital of Nanjing Medical University.