Background The hyperplasia of human Tenon capsule fibroblasts (HTFs) is a common cause of filtering surgery failure in glaucomous eye.Researches demonstrated that hydroxycamptothecin is a cell cycle arresting drug and induce apoptosis of cancer and fibroblasts.However,its mechanism is currently less understood.Objective This study was to investigate whether hydroxycamptothecin induce the apoptosis of HTFs and explore the possible mechanism.Methods Human Tenon capsule tissue was obtained from EyeBank of Jiangsu Province Hospital.HTFs were cultured using explant method in vivo and passaged in DMEM containing 10％ FBS.The cells were identified using vimentin and keratin by immunochemistry,and the cells of generation 3-6 in the logarithmic growth phase were used in the experiment.The cells were incubated with 0.01,0.05 or 0.10 g/L hydroxycamptothecin for 5 minutes respectively,and the cells without any hydroxycamptothecin were served as the control group.Cell viability then was assessed by cell counting kit-8 (CCK8) for the optimal inhibition concentration.The cells were treated by 0.10 g/L hydroxycamptothecin for 24 hours,and the apoptotic rate of the cells were assayed with annexin V/PI double-staining.Mitochondrial membrane potential of HTFs was assessed using JC-1 staining.The expressions of caspase-3,caspase-9 and cytochrome C (cyt C) in mitochondria and cytoplasm of HTFs were detected by Western blot.Results The proliferative value (A450) of the HTFs 0,0.01,0.05,0.10 g/L was 0.9716±0.0608,0.8035 ± 0.0346,0.7048 ±0.0446,0.6265 ±0.0286,with a significant difference (F =26.372,P =0.002).A450 of HTFs in the 0.01,0.05,0.10 g/L groups was significantly lower than the control group (P＜0.05),with the lowest A450 value in the 0.10 g/L group.The apoptotic percentage of HTFs was (18.72±1.41)％,in the 0.10 g/L hydroxycamptothecin group and that of the control group was (3.67 ±0.36)％,showing a significant difference between them (t =-10.374,P=0.001).The expression intensity of caspase-3 and caspase-9 protein in HTFs was higher in the 0.10 g/L hydroxycamptothecin group than that in the control group.JC-1 staining showed that the green fluorescence of the monomer JC-1 in cytoplasm was stronger in the 0.10 g/L hydroxycamptothecin group than that in the control group,but the red fluorescence of the polymer JC-1 in the 0.10 g/L hydroxycamptothecin group was weaker than that in the control group.The grey scale of cyt C protein in HTFs in mitochondrion was 0.0605±0.0022 in the 0.10 g/L hydroxycamptothecin group,showing a significant increase in comparison with 0.0301 ±0.0016 of the control group (t=4.865,P=0.014).However,the grey scale of cyt C protein in cytoplasm was declined in the 0.10 g/L hydroxycamptothecin group than that in the control group (0.0605 ±0.0022 vs.0.0301 ±0.0016) (t =-11.177,P =0.001).Conclusions Hydroxycamptothecin can induce the apoptosis of HTFs through activating the mitochondrial apoptosis pathway.

Objective To study the different effects between intermittent eliminating white blood cells and immunosuppressive agents pulse therapy in children′s anaphylatoid purpura nephritis on the base of hormone′s treatment.Methods Twelve cases of anaphylatoid purpura nephritis were enrolled in the study and divided into 2 groups.After failure in the hormone′s treatment,the observation group was applied with intermittent eliminating white blood cells(5 mL/kg,1-2 times/week),and the control group was treated with immunosuppressive agents[cyclophosphamide(CTX) 0.8-1.0 g/m~2,or methotrexate(MTX) 8-10 g/m~2].Results There were no significant differences between the 2 groups.The observation group had no obvious side effects,but the control group all had different side effects.Conclusion The treatment of intermittent eliminating white blood cells can have better effects without the side effect resulting from immunosuppressive agents.

Objective To investigate the influence of coxsackie B virus (CVB) and mycoplasma pneumoniae (MP) superinfection on immunological manifestation of recurrent respiratory tract inflammations(RRI) in children.Methods One hundred and thirty-two hospitalized children with RRI between Jan.to Dec.2005 were divided into negative control group,MP infection group,CVB infection group and superintection group by determining anti-MP IgM and anti-CVB IgM.Blood sedimentation,C-reactive protein,IgG,IgA,IgM and T lymphocyte subpopulation etc.were determined in four groups.The anti-MP IgM was determined by specificity immune agglutination test.Enzyme linked immunosorbent assay was used to detect anti-CVB IgM.The IgG,IgA,IgM were determined by simple agar diffusion method.T lymphocyte subpopulation was tested by flow cytometry.Results The percentage of CVB infection was 32.1%,mainly expressed in the 1-3 years old children;the percentage of MP infection was 22.7%,mainly expressed in children over 3 years old.In both CVB and MP infection group,the ten-dency of IgG increased and that of IgA decreased.IgM in the CVB and MP superinfection group was obviously higher than that in negative control group and MP infection group (Pa

Objective To investigate the dynamic changes of the spinal cord during neck flexion in Hirayama disease for diagnosis.Methods MRI examinations in neutral neck position and a fully flexed neck position were performed on 18 cases of Hirayama disease and 31 young normal control subjects.We measured an antero-posterior diameter(APD)and transverse diameter(TD)of the cervical cord at the superior margin of the C6 vertebral body for each position,and investigate the dynamic changes.The different in frequency of these findings between the control and patient groups was examined by means of the x~2 test.The group means were compared by independent-sample t-test.Significance was defined as P

Objective To analyze the impact of multiple factors on the incidence of pneumothorax associated with CT-guided transthoracic needle aspiration biopsy.Methods The sign of pneumothorax after 162 cases(lesion diameter from 1cm to 6cm)CT-guided transthoracic needle aspiration biopsy was observed and its relationship with multivariate factors were analyzed by multivariate logistic regression model.Results Thirty-two cases presented pneumothorax accounting for 19.8%.Single variate analysis showed that the sign of pneumothorax ralated to intercurrent COPD,distance from lesion and chest wall,needle dwelling time and lesion diameter.67 patients of intercurrent COPD with postoperative pneumothorax occurred in 22 cases (32.8%);With respect to those having lesions close to the chest wall(48 cases),and the cases with the distance between the chest wall and lesions less than 2cm(55 cases)and greater than 2cm(59 cases), the postoperative pneumothorax occurred in 0,14(25.5%),18(30.5%)cases respectively;For those patients with needle in the chest residence time of less than 10 minutes(82 cases),10—20 minutes (51 cases),more than 20 minutes(28 cases)after the occurrence of pneumothorax were 8(9.6%), 10(19.6%),14(50%)cases respectively;In contrast,those with lesion diameter less than 2 cm (65 cases),2—4cm(52 cases),more than 4cm(45 cases)were 19(29.2%),8(15.4%) and 5(11.1%)respectively.The multivariate logistic regression analysis showed that the prior three factors were risk factors of pneumothorax(OR=4.652,4.030,2.855 respectively).Conclusions To avoid the pneumothorax,caution must be taken with respect to CT-guided transthoracic needle aspiration biopsy, patients with intercurrent COPD,long distance between lesion and chest wall,and smaller lesion diameter. For operation the needle dwell time within thorax should be minimized.

Objective To evaluate the role of imaging diagnosis in surgical treatment of primary aldosteronism(PA). MethodsFrom Jan 1995 to Dec 2004,245 patients with PA were hospitalized in our hospital.Before the operations,all the patients underwent B-ultrasonography and CT scaning,240 received intravenous pyelography and 75 MRI.The preoperative imaging diagnosis were compared with the findings during the operations and postoperative pathologic results. Results Compared with the findings during the operations,the accuracy rates of localized diagnosis for PA with B-ultrasonography,CT scanning and MRI were 92.7%,98.2% and 90.4%,respectively.Compared with the postoperative pathologic results,the accuracy rates of qualitative diagnosis for aldosterone-producing adenoma(APA) with B-ultrasonography,CT scanning and MRI were 83.0%,90.7% and 72.2%,respectively. Conclusion The comprehensive imaging data are helpful in the localized diagnosis of PA.Correct preoperative qualitative diagnosis of APA is the key step for the surgical treatment for PA.

OBJECTIVETo explore the etiopathogenesis, therapy and incidence of pulmonary infection in kidney transplantation recipients taking new immunosuppressant.

METHODSThe clinical data from 752 kidney transplant recipients were retrospectively analyzed, who were divided into 3 groups according to the immunosuppressants administered, namely group A (CsA+MMF+Pred, n=226), group B (FK506+MMF+Pred, n=386) and group C (FK506+Rap+Pred, n=140). The incidence and mortality of pulmonary infection were recorded and the analysis of etiopathogenesis, diagnosis and therapy of pulmonary infection were carried out in the 3 groups.

RESULTSFifty-three patients acquired post-transplant pulmonary infection. The incidence of pulmonary infection was 7.08% (16/226) in group A, 7.25% (28/386) in group B and 6.43% (9/140) in group C. One patient died in group A and 2 in group B. Among the 53 patients, 24 had simple bacterial infection, 9 had cytomegalovirus infection, 1 had mycotic infection, 17 had combined infection, and 2 had unidentified pathogen infection. Of the pathogenic bacteria detected, 68.35% were Gram-negative.

CONCLUSIONGram-negative bacteria are most likely responsible for pulmonary infection after kidney transplantation, which most possibly occurs within 6 months after kidney transplantation. Early diagnosis and early treatment are critical for decreasing the mortality of severe pneumonia and for improving the survival rate of the patients and grafts.

Adolescent ; Adult ; Aged ; Cyclosporine ; administration & dosage ; adverse effects ; Cytomegalovirus Infections ; chemically induced ; diagnosis ; therapy ; Female ; Gram-Positive Bacterial Infections ; chemically induced ; diagnosis ; therapy ; Humans ; Immunosuppressive Agents ; administration & dosage ; adverse effects ; Kidney Transplantation ; Lung Diseases ; chemically induced ; diagnosis ; therapy ; Male ; Middle Aged ; Tacrolimus ; administration & dosage ; adverse effects

Ligustrazine, one of the major effective components of the Chinese traditional medicinal herb Ligusticum Chuanxiong Hort, has been reported plenty of biological activities, such as protect cardiovascular and cerebrovascular, neuroprotection and anti-tumor, et al. Because of its remarkable effects, studies on structural modification of ligustrazine have attracted much attention. Ligustrazine synthetic derivatives reported in recent decades are mainly derived from four primary intermediates (TMP-COOH, TMP-OH, TMP-NH2, HO-TMP-OH). To explore the neuroprotection activitiy of ligustrazine intermediates, six ligustrazine intermediates (2, 5, 8, 11, 12, 13) were synthesized and their protective effects against CoCl2-induced neurotoxicity in differentiated PC12 cells were studied. The target compounds were prepared via different chemical methods, including oxidation, substitution, esterification and amidation without changing the structure nucleus of ligustrazine. Compared with TMP (EC50 = 56.03 micromol x L(-1)), four compounds (2, 5, 12 and 13) exhibited higher activity (EC50 < 50 micromol x L(-1)) respectively, of which, compound 2 displayed the highest protective effect against the damaged PC12 cells (EC50 = 32.86 micromol x L(-1)), but target compounds 8 and 11 appeared lower activity (EC50 > 70 micromol x L(-1)). By structure-activity relationships analysis, the introduction of carboxyl, amino to the side chain of ligustrazine and appropriately increase the proportion of ligustrazine may contribute to enhance its neuroprotective activity, which provides a reference for the design, synthesis and activity screening of relevant series of ligustrazine derivatives in the future.
Animals ; Cell Differentiation ; drug effects ; Chemistry Techniques, Synthetic ; Cobalt ; toxicity ; Drugs, Chinese Herbal ; chemistry ; Neuroprotective Agents ; chemical synthesis ; chemistry ; pharmacology ; Neurotoxins ; toxicity ; PC12 Cells ; Pyrazines ; chemical synthesis ; chemistry ; pharmacology ; Rats

BACKGROUNDCerebral venous thrombosis (CVT) may lead to serious neurological disorders; however, little is known about the risk factors for recurrent CVT. Our aim was to determine the association between elevated fibrinogen and decreased high-density lipoprotein cholesterol (HDL-C) levels with recurrent CVT.

METHODSThis retrospective cohort study included participants if they had a first episode of objectively defined CVT and were admitted to Xuan Wu Hospital, Capital Medical University from August 2005 to September 2009. Demographic and clinical variables were collected, as well as laboratory parameters, including plasma fibrinogen and HDL-C. Patients with CVT were followed for recurrent symptomatic CVT. Follow-up was through the end of September 2010. Potential predictors of recurrence were analyzed using Cox survival analysis.

RESULTSAt the end of the follow-up, 95 patients were eligible for the study. Twelve of 95 patients (12.6%) had recurred CVT. The median time of recurrence was 7 months (range: 1-39 months). Eight of these 12 (66.7%) experienced recurrence within the first 12 months after their initial CVT. The recurrence rate of CVT was 2.76 per 100 patient-years. Multivariate Cox regression analysis demonstrated that the coexistence of high fibrinogen (>4.00 g/L) and low HDL-C (<1.08 mmol/L) levels at baseline was the only independent predictor for recurrent CVT (hazard ratio: 4.69; 95% confidence interval: 1.10-20.11; P < 0.05). Of the twelve patients with recurrent CVT in our study, 7 (58.3%) had high fibrinogen plus low HDL-C levels. All 7 of these patients took warfarin for 3-12 months, and 6 of 7 had recurrent CVT after the discontinuation of anticoagulant treatment.

CONCLUSIONSConcomitant high fibrinogen and low HDL-C levels may be associated with recurrence of CVT. The effect of potential risk factors related to atherothrombosis on recurrent CVT should be closely monitored.

Adolescent ; Adult ; Aged ; Cholesterol, HDL ; metabolism ; Cohort Studies ; Female ; Fibrinogen ; metabolism ; Humans ; Intracranial Thrombosis ; metabolism ; pathology ; Male ; Middle Aged ; Recurrence ; Retrospective Studies ; Young Adult

OBJECTIVETo investigate the inhibition mechanisms of BAY11-7082 (IkappaB-alpha phosphorylation inhibitor) and Lactacystein (proteosome inhibitor) in CD154-induced NF-kappaB activation.

METHODSWe used recombinant CD154 to stimulate EBV/LMP1 negative Ramos B cell and observed the effects of BAY11-7082 and Lactacystein in CD154-induced NF-kappaB luciferase activation, phosphorylation and degradation of IkappaB-alpha, phosphorylation of p65, and nuclear translocation of NF-kappaB subunits upon CD154 stimulation.

RESULTSBoth BAY11-7082 and Lactacystein abrogated CD154-induced NF-kappaB luciferase activation in Ramos cells. While CD154-induced phosphorylation of p65, phosphorylation and degradation of IkappaB-alpha, and nuclear translocation of p50, p65, and c-Rel were all blocked by BAY11-7082; Lactacystein only inhibited degradation of IkappaB-alpha and p65 nuclear translocation.

CONCLUSIONBAY11-7082 and Lactacystein inhibit CD154-induced NF-kappaB activation through different mechanisms.

Acetylcysteine ; analogs & derivatives ; pharmacology ; Apoptosis ; drug effects ; Burkitt Lymphoma ; pathology ; CD40 Ligand ; pharmacology ; Cysteine Proteinase Inhibitors ; pharmacology ; Enzyme Activation ; drug effects ; Humans ; NF-kappa B ; antagonists & inhibitors ; metabolism ; Nitriles ; pharmacology ; Sulfones ; pharmacology ; Tumor Cells, Cultured