Benzene ; toxicity ; Carcinogens ; toxicity ; Chromosome Aberrations ; Genetic Predisposition to Disease ; Humans ; Leukemia ; chemically induced ; genetics

BACKGROUND:Cauda equina syndrome often induces skin hypoesthesia in the perineal area, poor urine-stool control, and impairs male function. After peripheral nerve fiber injury, apoptosis of neurons appeared. This is associated with the nature of the injury, the types of neurons, the species of animals, the age, and the distance between neurons. OBJECTIVE:To explore the motor neuron apoptosis and expression of apoptosis-associated protein in the anterior horn of the spinal cord after acute cauda equina compression. METHODS:A total of 27 canines were randomly divided into three groups. In the compression and control groups, models of cauda equina compression were established. In the normal group, no models were established. Compression group received water sac compression for 4, 8, 12, 24, 48, 72 and 168 hours, with three models in each group. In the control group, only water sac was implanted, but water was not injected. Terminal deoxynucleotidyl transferase TdT-mediated biotin dUTP nick end-labeling assay was used to detect the apoptosis of neurons in the anterior horn of the spinal cord. Bcl-2, Bax and Caspase-3 protein expressions were measured by immunohistochemical staining (strept avidin-biotin complex). Gray values of positive cels of Bax, Bcl-2 and Caspase-3 protein expressions were detected using Qwin550Cw image colection and analysis system. RESULTS AND CONCLUSION:The apoptosis of motor neuron occurred in the compression groups. At 12 hours of compression, positive cels were detected, and the number of positive cels reached a peak at 72 hours. Bax and Bcl-2 protein expression was smal in the normal group. Caspase-3 protein expression was not detected in the normal and control groups. Bax and Bcl-2 protein expression was significantly increased at 8 hours, peaked at 72 hours and reduced to a normal level at 168 hours. The increased range of Bax protein expression was bigger than that of Bcl-2. Caspase-3 protein began to express at 12 hours, peaked at 72 hours and reduced to a low level at 168 hours. Bax and Caspase-3 protein expression peaked at 72 hours, and Bcl-2 protein expression was not obviously increased. These findings verified that after acute cauda equina compression, the apoptosis of neurons occurred in the anterior horn of the spinal cord. Bax and Bcl-2 protein expression showed an antagonistic action. In the Bax/Bcl-2 complex, Bax protein in a high expression promoted apoptosis, induced Caspase-3 protein expression, and neuronal apoptosis.

· AIM :To investigate the role of pericytes in growth of retinal microvascular endothelial cells with a co-culture system in order to understand some mechanism of angiogenesis in hypoxia induced retinal neovascular disorders.(RMECs) were isolated by a modified protocol using CD31 coated Dynabeads, and identified by immunocytochemical staining with anti-Factor Ⅷ and CD31 antibodies. Rat retinal pericytes were isolated and characterized by immunofluorescent staining with PDGFR-β; and desmin antibodies. Pericytes and RMECs were cultured in a contact co-culture system both under normoxia and hypoxia by Millicell chamber. RMECs proliferation was evaluated by MTT and cell cycle assay with flow cytometry. RT-PCR was used to detect the alteration of KDR/Flk-1 mRNA level in RMECs under normoxia or hypoxia in the co-culture system.harvested with the modified isolating method. The two cell types were identified by positive Factor Ⅷ, CD31 and PDGFR-β, desmin cytochemical staining respectively.RMECs proliferated significantly under hypoxia from 3 to 9d with a maximal rate on day 6 (24.9%, P ＜ 0.01) by MTT. In the co-culture system, the proliferation of RMECs was inhibited by pericytes. After 6d exposure to hypoxia,the fraction of S-phase RMECs number was greatly increased by 43.9% (P ＜ 0.01). In the co-culture system,RMECs proliferation was inhibited by pericytes through decreasing the fraction of S-phase cell number both under normoxia (3.6%, P＜0.05) and under hypoxia (15.1%,P＜0.01). KDR/Flk-1 mRNA level in single cultured RMECs was shown to increase approximately 1.3-fold when exposed to hypoxia. Compared with single cultured RMECs, co-culture with pericytes could decrease KDR/Flk-1 mRNA by 45.1% (P＜0.05) and 27.7% (P ＜ 0.05) under normoxia and hypoxia condition respectively.pericytes could inhibit proliferation of RMECs under both normoxia and hypoxia. The inhibition effects of pericytes maybe, at least in part, due to downregulation of KDR/Flk-1 of RMECs. These findings confirm that pericytes could be a potential inhibitor in the pathogenesis of retinal neovascularization.

OBJECTIVETo evaluate the effect of Tongnao Huoluo Acupuncture (THA) therapy on acute cerebral infarction (ACI).

METHODSAdopting multi-centered, randomized and controlled method, 397 ACI patients from 10 hospitals subjected to the study were treated according to the initiating time (IT) of disease during hospitalization: the 138 patients of stage-1 with IT < or =6 h, were randomly assigned to three groups, treated respectively with THA (Group A), thrombolysis with urokinase (Group B) and Batroxobin (Group C); the 140 patients of stage-2 with IT within 6-48 h, and 119 patients of stage-3 with IT within 48 h-14 d were randomly assigned to three groups, treated respectively with THA (Group D) body acupuncture (Group E), and conventional treatment (Group F). Therapeutic effect was evaluated by NIHSS scores estimated at the day 1, 3, 7, 14, 28 and 90 of treatment, and the Barthel Index (BI) measured at day 14, 28 and 90.

RESULTSEffect in Group A was insignificantly different from that in Group B (P > 0.05), but was different from that in Group C significantly (P < 0.01). At day 90, the percentage of patients with high BI (HBI%, patients with BI >95%) was insignificantly different in Group A vs. B (P > 0.05), but was significantly different in Group A vs. C (P < 0.01). Comparisons between Group D, E and F showed that the therapeutic effect in Group D was equivalent to that in Group E (P < 0.05), but better than that in Group F (P < 0.01), and HBI% in Group D was superior than that in the other two groups (P < 0.01).

CONCLUSIONSTHA therapy shows favorable effects in reducing the crippling rate and improving the living capacity of ACI patients.

Acupuncture Therapy ; methods ; Adult ; Aged ; Cerebral Infarction ; therapy ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome

Objective To analyze the local control rate and the desimetric patterns of local recurrence in nasopharyngeal carcinoma(NPC)patients having been treated with standardized conventional radiotherapy and to evaluate the delineation of rational target volume.Methods From Jan.2000 to Dec.2000,476 patients with untreated NPC were treated by standardized conventional radiotherapy alone at the Sun Yat-sen University Cancer Center.The radiation ports were designed on a X-ray simulator.The nasopharyngeal lesion demonstrated by CT scan and the subclinical spread regions adjacent to the nasopharynx were defined as the target volume.Kaplan- Meier method was used to calculate the cumulative local recurrence rate.For patients with locad recurrence,the primary and recurrent local tumor volumes(V_(nx),V_(recur))were delineated with three-dimensional treatment planning system(3DTPS),and the dataset of radiation ports and delivered prescription dose to the 3DTPS were transferred according to the first treatment.The dose of radiation received by V_(recur)was calculated and analyzed with dose- volume histogram(DVH).Local recurrence was classified as:1.“in-port”with 95% or mere of the recurrence volume((recur)_V_(95))was within the 95% isedase;2.“marginal”with 20% to95% of _(recur)V_(95)within the 95% isedese; 3.“outside”with only less than 20% of _(recur)_V_(95)within the 95% isodose curve.Results With the median follow- up of 42.5 months(range 8～54 months),52 patients developed local recurrence.The 1-,2-,3 and 4-year cumulative local failure rate was 0.6%,3.9%,8.7% and 11.5%,respectively.Among the 42 local recurrent patients who could be analyzed by 3DTPS,52% were in-port,40% were marginal and 7% were outside.For most of the marginal recurrence and all the outside recurrence patients,the main reason of recurrence were related to the unreasonable design of the radiation port and inaccuracy in the interpretation image findings.Conclusions The outcome of better local control rate and the dosimetric pattern of local recurrence show that the target volume is reasonable for NPC in Sun Yat-sen University Cancer Center.Enhancing the capability of correct interpretation of images,accurate design of the radiation pouts and making most useful molecular or functional imaging techniques to escalate the local radiation dose are promising ways to improve the local control further and better.

Objective This study is to investigate the set-up accuracy of thermoplastic mask used for immobilization of nasopharyngeal carcinoma (NPC) patients being treated by Intensity Modulated Radia- tion Therapy (IMRT).Methods Nineteen patients with early stage nasopharyngeal carcinoma (T1- T2N0M0),treated by fractionated intensity-modulated radiation therapy,underwent repeated CTs during their 6-week treatment course.We evaluated their anatomic landmark coordinates in a total of 85 repeated CT data sets and respective x,y and z shifts relative to their position in the 19 treatment-planning reference CTs.Results The translation in x,y,and z-axes with their mean value derived from both positive and negative set-up errors was-0.84 mm(x-axis),+0.65 mm(y-axis) and +0.01 mm(z-axis).Mean target isocenter translation was (0.89?0.69) mm,(0.82?0.79) mm,(0.95?1.24) mm in x,y and z-direc- tions,respectively.The mean total magnitude vector and 95% CI of isocenter motion were 1.87 mm and 4.65 mm.The data measured over the 6-week fractionated course of treatment revealed a slight deterioration of isocenter accuracy.The 95% CI,considered by us to be a valuable parameter for characterizing the sys- tem,of 4.17 mm for measurement within the first 3 weeks increased to 5.12 mm in the last 3 weeks of treat- ment.Conclusions The sequential CT scanning is a pronounced valuable method of evaluating the quality of departmental specific patient positioning procedures.The thermoplastic mask is eyed as well suited tool for immobilization and repositioning of NPC patients receiving intensity-modulated radiation therapy.

OBJECTIVETo explore the association of the glycophorin A(GPA) gene mutation in peripheral erythrocytes and chronic benzene poisoning.

METHODSSixty-three patients with chronic benzene poisonings and 45 benzene-exposed workers who were engaged in the same job title were investigated. Fluorescence immunolabeling technique and flow cytometry were used to detect GPA mutation frequency in peripheral read blood cell.

RESULTSA significant decrease in WBC count and neutrophil count was found in patients with chronic benzene poisoning compared with control individuals (P<0.01). The WBC count and neutrophil count both decreased along with the GPA-NN frequency, and the trends were significant(P<0.05).Both WBC counts and neutrophil counts decreased as the frequency, and trends were significant(P<0.05). GPA-NN frequency increased along with the accumulative exposure score, and the trend was significant (P = 0.0026). There was no significant trend between the GPA-Nphi frequency and the accumulative exposure score (P = 0.2037).

CONCLUSIONA decrease in WBC count and neutrophil count is found in patients with chronic benzene poisoning, which can arise from genetic damage in bone marrow stem cells, namely gene-duplicating mutations (NN) at the GPA locus in bone marrow cells of MN-heterozygous subjects, GPA-NN mutagens contributed to the pathogenesis of chronic benzene poisoning.

Adult ; Benzene ; poisoning ; Bone Marrow Cells ; pathology ; Case-Control Studies ; Erythrocytes ; pathology ; Female ; Genetic Variation ; Glycophorin ; genetics ; Humans ; Leukocyte Count ; Male ; Middle Aged ; Mutation Rate ; Neutrophils ; pathology

AIMTo study the direct effect and kinetics of sodium quercetin-7,4'-disulphate (SQDS) on recombinant human protein kinase CK2 holoenzyme.

METHODSThe recombinant human CK2 holoenzyme activity was assayed by detecting incorporation of 32P of [gamma-32P] ATP into the substrate in various conditions.

RESULTSThe recombinant human CK2 was a second messenger (Ca2+, cAMP and cGMP) independent protein kinase. The characterization and function of the reconstituted holoenzyme were consistent with those of native CK2. SQDS was shown to strongly inhibit the holoenzyme activity of recombinant human protein kinase CK2 with an IC50 of 4.4 mumol.L-1, which was more effective than DRB and A3, known CK2 special inhibitors. Kinetic studies of SQDS on recombinant human CK2 showed: the inhibition was competitive with ATP and noncompetitive with casein.

CONCLUSIONSQDS is a potent inhibitor of protein kinase CK2. This study provide experimental basis for the development of more effective inhibitors of CK2 and for clinical application of SQDS in the future.

Casein Kinase II ; Dichlororibofuranosylbenzimidazole ; pharmacology ; Enzyme Inhibitors ; pharmacology ; Humans ; Kinetics ; Protein-Serine-Threonine Kinases ; antagonists & inhibitors ; metabolism ; Quercetin ; analogs & derivatives ; pharmacology ; Recombinant Proteins ; antagonists & inhibitors ; metabolism

OBJECTIVETo study a new method of venous flap that is improved on its persistence and quality.

METHODSNew Zealand white rabbits were subdivided randomly into 4 groups. All rabbits were operated by harvesting a flap from the latero-abdominal wall and then sutured it in the original position. Group A: the superficial epigastric vein in the pedicle was left open (only one inflow vein remained). Group B: the pedicle vein of the proximal and distant end were left open (keeping an inflow vein and a principal out). Group C: the pedicle vein and a tributary vein were left open (keeping an inflow vein and a tributary outflow vein). Group D: the pedicle vein and two tributary veins were left open (keeping an inflow vein and two tributary outflow veins). Survival rate, MDA of the tissue, histology and ultra-microstructure were examined.

RESULTSSurvival rate of A, B, C, D were improved in order. Statistic difference is significant (P < 0.05) between group and group other than C and D. The content of MDA was heightened with statistically significant differences (A > B > C > D) among the four groups 8 hours postoperatively, but fell back to the normal level in group D and C and kept a high level in group A and B at 72 hour postoperatively. Histology and ultra-microstructure exam showed that degeneration of collagen fiber and karyopyknosis of cell is more obvious in Group A and Group B than Group C and Group D.

CONCLUSIONSThe higher survival rate of venous flap is possible by designing the more reasonable venous flap outputs pedicles which can alleviate the high tension dropsy and maintain the valid equilibrium of pour with flow in the venous flap.

Animals ; Graft Survival ; Rabbits ; Surgical Flaps ; blood supply ; Veins ; transplantation

Objective To analyse the preliminary clinical results of intensity modulated radiation therapy (IMRT) for 122 untreated nasopharyngeal carcinoma (NPC)pafients.Methods 122 NPC pa- tients received IMRT alone from Feb.2001 to Jun.2004,with 31 females and 91 males,and a median age of 45 years(range 25-66).According to the Fuzhou Stage Classification,there were StageⅠ11 patients, StageⅡ34,StageⅢ62,and StageⅣa 15.IMRT was carried out using an inverse planning system (COR- VUS 5.0,Peacock plan) developed by the NOMOS Corp.The treatment was given with the Multi-leaf Inten- sity Modulating Collimator (MIMIC) using a slice-by-slice arc rotation approach.The prescription dose was 68 Gy/30f to the nasopharynx gross tumor volume (GTV_(nx)),60-66 Gy/30f to positive neck lymph nodes (GTV_(nd)),60 Gy/30f to the first clinical target volume (CTV_1) and 54 Gy/30f to the second clinical target volume (CTV_2).Kaplan-Meier method was used to calculate the overall survival rate (OS),distant metas- tasis-free survival rates (DMFS),and local-regional control rates from the last date of therapy.Log-rank test was used to detect the difference between groups.Results The median follow-up time was 20 months ( range 6 to46 months).The 1-,2-,and 3-year OS was 95.2%,91.4%,85.1%,DMFS was 91.9%, 88.6%,85.6%,and the local-regional control rates was 96.5%,93.2%,93.2%,respectively.Statistics of the local control rate was insignificant either for advanced T(T3+T4) stage or early T(T1+T2) stage diseases(P=0.148).The 2-year regional control rate was insignificant either for patients with N(+) or N (-),but the 2-year DMFS was significant both for patients with N(+) and N(-)lesions(P=0.004).For 17 patients who failed,there were two with residual disease and one with recurrence at the primary site (17.6%),three patients in the neck (17.6%),twelve patients (70.6%) in distant metastases.Conclu- sions Intensity modulated radiation therapy does provide excellent local-regional control for untreated NPC, especially in patients with advanced T stage or N(+) lesion.Distant metastasis is the main cause of failure. N (+) is significantly correlated with distant metastasis.