Objective:To observe the changes in differentiation,development and function of DCs after mouse bone marrow-derived dendritic cells devouring autoantigen in vitro,to study its impact on the immune status.Methods:Mouse bone marrow cells were cultured in GM-CSF+IL-4 to generate DCs,and the purity of DCs was detected by fluorescence microscope and flow cytometry.It was identified whether DCs could swallow autoantigen by immunohistochemistry and transmission electron microscope;and the changes in CD86,MHCⅡ on DCs after swallowing autoantigen were observed by flow cytometry.The changes in proliferation after the mouse spleen lymphocytes were cultured with the same genetic background of DCs were measured by MTT.The amounts of IL-4,IFN-? secreted by the mouse spleen lymphocytes after stimulation with DCs were measured by ELISA.Results:The purity of DCs was 90.6%.DCs could swallow autoantigen,which induced differentiation dysplasia of the DCs in vitro.Swallowing autoantigen had negative effect on the maturation as well as the potentiation to stimulate proliferation of the mouse spleen lymphocytes with the same genetic background.Secretion of IFN-? by stimulated lymphocytes decreased,but secretion of IL-4 promoted.Conclusion:The DCs that swallow autoantigen could inhibit imDCs maturation,but promote the immune tolerance and humoral immunity.

Objective To study the expressions of PTTG and bFGF proteins and their relationship with microvessels density(MVD)in esophageal carcinoma.Methods Immunohistochemical SP method was used to detect the expression of PTTG and bFGF proteins in 48 esophageal carcinoma tissues and the same para-cancerous tissues.MVD was evaluated by immunohistochemieal staining with antibody CD34.Results The positive rate of PTTG and bFGF was 68.8%(33/48)and 70.8%(34/48)respectively.Rate of PTTG protein expression in esophageal carcinoma tissues was significantly higher than that in para-cancerous tissues(8.3%and 12.5%,P＜0.05).The positive rate 0f PTTG,bFGF and MVD was correlated with lymph node metastasis and TNM stage.There was no relationship with age,sex,tumor size MVD(P＜0.05).Conclusion PTTG and bFGF are over-expressed in esophageal carcinoma.Increased PTTG may play an important role in carcinogenesis and development of esophageal carcinoma by promoting the expression of bFGF protein which may induce an angiogenesis.

Aged ; Carcinoma, Hepatocellular ; diagnostic imaging ; pathology ; surgery ; Hepatectomy ; methods ; Humans ; Liver Neoplasms ; diagnostic imaging ; pathology ; surgery ; Male ; Middle Aged ; Tomography, X-Ray Computed

Erythromelalgia ; complications ; Female ; Humans ; Mercury Poisoning ; complications ; Middle Aged

Background Research showed that transforming growth factor-β2 (TGF-β2) promotes scar formation.But its mechanism in scarring after glaucoma filtration surgery is worthy of studying.Objective This study was to investigate the effect of TGF-β2 on myofibroblast transition of human Tenon fibroblasts (HTFs) and scarring after glaucoma filtration surgery.Methods Tenon capsular tissue was obtained from 3 patients with strabismus during the surgery and was incubated in DMEM with 10％ fetal bovine serum (FBS).The cells were collected and passaged in the free-serum medium for 24 hours,and then 1,2,5,10,20 μg/L TGF-β2 was added into the medium respectively,to induce the transformation of HTFs,and 2 μg/L or 5 μg/L TGF-β2 was used to treat the HTFs for 6,24,48 and 72 hours.The control group was not treated with TGF-β2.The expressions of α-smooth muscle actin (α-SMA) and phosphorylation of the signaling proteins (pSmad2) in HTFs were detected by Western blot assay.The expressions of α-SMA and F-actin were located by cell immunofluorescine technique under the confocal immunofluorescence microscopy.Cell contractility was determined by collagen gel contraction assays.This study was approved by Ethic Committee of Institute of Surgery Research of Daping Hospital,and informed consent was obtained from each patient or custodian initial of the study.Results The expression of α-SMA protein in the HTFs was increased significantly after the treatment of TGF-β2 in comparison with the control group and reached a peak at 24-48 hours.The α-SMA expression was gradually weakened in the 10 μg/L TGF-β2 groups.Little of α-SMA and F-actin were expressed in the control group.However,strong staining for α-SMA and F-actin were observed in the 1,2 and 5 μg/L TGF-β2 groups and then the staining weakened at the concentration of 10 μg/L.In addition,pSmad2 showed a stronger expression in the 2 μg/L TGF-β2 group than that in the PBS group and FBS group,with the strongest expression in 30 minutes through 2 hours.The untreated gel contracted (78.00±3.13)％ from its initial size,and contraction in the 1,2,5,10 μg/L TGF-β2 group were (63.88±1.78)％,(20.69±0.65)％,(19.49-±0.54)％,(16.24±0.84) ％,respectively,TGF-β2 increased HTFs contraction significantly (Fgroup =859.400,P =0.000).Conclusions TGF-β2 can induce transdifferentiation of Tenon fibroblast into myofibroblast and increase cell contractility,with a concentration-dependent and time-dependent pattern to an extent.It may be the mechanism of scar formation after glaucoma filter surgery.

Objective To evaluate the long-term efficacy of recombinant human endostatin (rh-Endo) combined with FOLFOX4 as an adjuvant treatment for patients of stage Ⅱ and Ⅲ colorectal cancer.Methods Eligible patients were randomly assigned to receive FOLFOX4 or FOLFOX4 plus rh-Endo regimen in which patients receiving 7.5 mg/m2 Ⅳ on day 1-7,repeated every 2 weeks,to a total of 12 cycles in 6 months.Results A total of 197 eligible patients were accrued in this research with 105 patients in the control group and 92 patients in the experimental arm.Median follow-up period was 42 months.The baseline characteristics distributed were balanced by treatment.Rh-Endo combined with FOLFOX4 regimen resulted in significant improvement on DFS compared to FOLFOX4 regimen for patients with stage Ⅲ colon cancer (HR =0.19,95％ CI0.05-0.75,P =0.0124),and with a 34％ improvement on 3-year DFS and 81％ reduced recurrence.Although rh-Endo combined with FOLFOX4 regimen failed to make significant difference on DFS in the whole (HR =0.75,95％ CI 0.31-1.83,P =0.5589),it was also observed a 17％ improveiment on 3-year DFS.No statistical significant difference on DFS was observed in patients with stage Ⅱ disease.Conclusions Rh-Endo combined with FOLFOX4 regimen significantly improved the disease-free survival for patients with stage Ⅲ colorectal cancer,indicating that patients with stage Ⅲ disease,but not stage Ⅱ disease,can benefit from FOLFOX4 plus rh-Endo regimen in adjuvant treatment.

The IL-8 and MMP-7 genes participate in the carcinogenesis of many malignancies, but the role of both genes in cervical cancer is not fully elucidated. The aim of this study was to determine the frequency of IL-8 and MMP-7 gene mutations and to assess their effects on the risk of early stage cervical cancer and lymph node metastasis. The clinical stage and histological grade of cervical cancer were also studied. The peripheral blood from the patients with early stage cervical cancers and normal controls was collected and the DNA was extracted. The incidence of IL-8 and MMP-7 gene mutations was assessed by using tetra-primer amplification refractory mutation system polymerase chain reaction (ARMS PCR) and restriction fragment length polymorphism (RFLP). The data were statistically analyzed by x2 test. The results showed that: (1) The genotype frequency of IL-8 -251AT and TT was significantly higher in the cervical cancer group than in the normal control group (OR=2.290 and 2.619 respectively, P=0.001), and it was also higher in the lymphatic metastasis group than that without metastasis (OR=2.917, P=0.035); (2) The frequency of MMP-7 -181G/G genotype was significantly higher in the cervical cancer group and in the lymphatic metastasis group (P<0.05); (3) The incidence of IL-8 mutation was two times higher in IIa cervical cancer group than in Ib1 and Ib2 cervical cancer group (P=0.006). For the MMP-7 gene, there was statistically significant difference in the incidence of mutation between the Ib1, Ib2 and the IIa (P=0.000); (4) Different histological types and different grades of cervical cancer had different incidence of mutations, statistically. It was suggested that there was significant difference in the genotype of IL-8 -251TT and MMP-7 -181GG polymorphism between the cervical cancer group and the lymph node metastasis group. Moreover, individuals with IL-8 T allele or MMP-7 G allele carriers were at significantly higher risk of cervical cancer, particularly the early (IIa) and medium, poorly differentiated cervical cancer (G2+G3).

Background Our previous study demonstrated that the aptamer S58 specifically targeted transforming growth factor-β receptor Ⅱ (TβRⅡ) and inhibited the transdifferentiation of human Tenon capsule fibroblasts (HTFs) mediated by transforming growth factor-β (TGF-β).Chitosan-nanoparticles (CS-NP) are good drug carriers,but the efficacy and safety of CS-NP/aptamer complexes deserve attention.Objective The aim of this study was to synthesize a novel CS-NP/aptamer complex called CS (S58)-NP and investigate its properties and applicability.Methods Human Tenon capsule tissue was obtained from patients during strabismus surgery,and HTFs were cultured and passaged using the explant culture method.The fourth to tenth generations of cells were used in the experiment.Different concentrations of CS-NP were used to prepare the CS(S58)-NP by the ionic cross-linking method with a surface charge rate (N/P) for S58 of 10,20,30 or 40.The particle size and Zeta potential were measured by the Zeta analyzer.The shape and distribution of CS (S58)-NP particles were examined under the scanning electron microscope.The binding of CS-NP with S58 and resistance of CS (S58)-NP to DNase Ⅰ were examined by agarose gel eletrophoresis.The release rate of S58 from CS (S58)-NP in PBS was quantitatively analyzed by a ultraviolet spectrophotometer.The cytotoxicity of CS(S58)-NP to HTFs was evaluated by detecting the production of lactate dehydrogenase (LDH).Results The Zeta analyzer showed that the particle size of CS (S58)-NP was 130-270 nm and its electric potential ranged from + 16 to +28 mV.The CS (S58)-NP particles appeared spherical with an even distribution under the scanning electron microscope.The mean encapsulation efficiency of CS(S58)-NP was 88.9％,89.3％,91.7％ or 90.5％,respectively,when the N/P was 10,20,30 or 40.After being encapsuled by CS-NP,S58 could resist the degradation from DNase I.Its total releasing level in PBS increased with the lapse of time,with a maximum releasing speed at 24 to 36 hours.The total releasing level reached 100％ at 96 hours.With increaseing concentrations of CS(S58)-NP,the relative releasing level of LDH in HTFs suspension gradually elevated with a significant difference among the groups (F =588.018,P =0.000),with the highest released LDH level at 50 nmol/L of CS(S58)-NP (12.853％ ±0.375％).Conclusions CS-NP provides a protective and slow-releasing effect on the S58 aptamer.CS (S58)-NP shows a good biocompatibility with HTFs with a low cytotoxicity at a concentration of ＜50 nmol/L.CS(S58)-NP could be used to inhibit TGF-β induced transdifferentiation of HTFs in the future.

BACKGROUND:Percutaneous pedicle screw fixation has been applied in the treatment of thoracolumbar fracture,and has achieved satisfactory clinical efficacy.Injectable calcium sulfate holds good biocompatibility,degradability and fast curing.OBJECTIVE:To explore the clinical efficacy of percutaneous pedicle screw fixation combined with calcium sulfate cement for single-level thoracolumbar fracture.METHODS:Clinical data of 40 patients with single-level thoracolumbar fracture without nerve injury were analyzed retrospectively.All fractured vertebrae were compressed more than 30％ and at least one pedicle was complete.All patients were treated with percutaneous pedicle screw fixation combined with calcium sulfate cement.The Visual Analog Scale and Oswestry Disability Index scores were recorded at baseline,3 months postoperatively and last follow-up;the height of the fractured vertebra body and sagittal Cobb angle were measured on X-ray;the patient's satisfaction and healing rate were recorded.RESULTS AND CONCLUSION:(1) The Oswestry Disability Index at 3 months postoperatively and last follow-up was 16.3％ and 4.4％,respectively.Compared with baseline,the Cobb angle and Visual Analog Scale scores were significantly reduced,and height of the fractured vertebra body was significantly increased after surgery (P ＜ 0.01).(3) The healing rate at last follow-up was 95％,nonunion was not found,and the patients' satisfaction reached 95％.(4) The loss of vertebral height and Cobb angle was found at last follow-up compared with 3 months postoperatively,but had no significant difference (P ＞ 0.05).(5) These results indicate that percutaneous pedicle screw fixation combined with calcium sulfate cement is safe and reliable for single-level thoraclumbar fracture,which not only restores the vertebral height and relieves pain,but also has satisfactory long-term curative efficacy and high healing rate.

Objective To explore the effect of Okam on airway inflammation in asthmatic mouse.Methods Thirty-two SPF grade Kunming Strain mice were randomly divided into positive control group,glucocorticoid inhalation group,Okam group and negative control group with 8 mice in each group.The mice were sensitized and repeatedly challenged with ovalbumin(OVA) to establish the models of chronic asthma.The glucocorticoid group were given Budesonide(200 ?g) and saline everyday by inhalation,the Okam group were given 50 mg/kg Okam by gavage,and the positive group had saline at the same time,the negative control group received saline at all stages.The inflammation of the lung tissue were scored underwent HE staining.Bronchoalveolar lavage fluid(BALF) cell count and differential were studied,and interferon-?(IFN-?),interleukin-4(IL-4) in BALF were determined by enzyme-linked immunosorbent assay(ELISA).Results There were no inflammatory cell infiltrate of bronchiole in the negative control group.Inflammatory infiltration of lung tissue were obvious in the positive control group.Inflammatory infiltration of lung tissue lightened obviously in the Budesonide and Okam groups.The total cell number,Eosinophils(EOS) and IL-4 level in BALF,and the score of the lung tissue in Okam group were all markedly lower than those in positive control group(t=5.942,7.089,7.078 Pa0.05),IFN-? lower(t=4.275 P