Objective To study the effect of TGF-?1 on the activation of ERK MAPK in human bronchial epithelial BEP2D cells. Methods Western blot was employed to examine the time-dependent activation of ERK MAPK by TGF-?1. BEP2D cells were harvested after treatment of human bronchial epithelial cells with 2 ng/ml TGF-?1 for 0, 10, 30, 60, 120, 240 and 480 min, respectively. Fluorescent dye staining and flow cytometry were employed to assess the apoptosis of BEP2D cells treated with vehicle, or with 2ng/ml TGF-?1, or co-treated with 2ng/ml TGF-?1 and 5?M U0126. Proliferation of BEP2D cells treated with vehicle, or with 2ng/ml TGF-?1 or 5?M U0126, or co-treated with 2ng/ml TGF-?1 and 5?M U0126 was assayed with colony-forming test, respectively. Morphological observation was performed to observe the morphological changes in BEP2D cells treated with vehicle, or with 5ng/ml TGF-?1 or 5?M U0126, or co-treated with 5ng/ml TGF-?1 and 5?M U0126, respectively. Results TGF-?1 activated ERK MAPK in BEP2D cell. The maximal activation of ERK MAPK took place at 60min after stimulation with 2ng/ml TGF-?1. TGF-?1 treatment effectively inhibited cell proliferation, and induced their apoptosis and epithelial-mesenchymal transition. Pretreatment with U0126, an inhibitor of ERK MAPK, significantly enhanced the TGF-?1-mediated anti-proliferation and apoptosis effects, and inhibited the effect of epithelial-mesenchymal transition of TGF-?1 in BEP2D cells. Conclusion TGF-?1-induced phosphorylation of ERK MAPK may participate in BEP2D cell proliferation and apoptosis regulation.

Objective To investigate the mechanism of malignant transformation in human bronchial epithelial cell line BEP2D exposed to α-particles.Methods The levels of intracellular ROS and malonaldehyde (MDA) in BEP2D,RH22 (passage 22 of α-particle-irradiated BEP2D cells) and BERP35T-1 cells (derived from nude mice bearing malignant transformed cells generated from the passage 35 of α-particle-irradiated BEP2D cells) were assayed with DCFH-DA and MDA kit,respectively.The expressions of 8-OH-dG and γ-H2AX in BEP2D,RH23 (passage 23 of α-particle-irradiated BEP2D cells)and BERP35T-1 cells were also measured with immunocytochemistry and immunofluorescence staining.Results Compared to BEP2D cells,the levels of ROS ( t =4.30 and 3.94,P ＜ 0.05 ) and MDA ( t =4.89 and 15.10,P ＜0.05) increased in RH22 and BERP35T-1 cells.The expressions of 8-OH-dG (t =3.80 and 2.92,P ＜ 0.05 ) and γ-H2AX ( t =7.61 and 12.67,P ＜ 0.05 ) in RH23 and BERP35T-1 cells were also higher than those in BEP2D cells.Conclusions Oxidative stress induces lipid peroxidation and DNA damage leading to genomic instability,which could contribute to cellular malignant transforming process in the human bronchial epithelial cell line BEP2D with α-particle exposure.

Objective To investigate the antioxidant ability and radiosensitivity in the malignant transformed human bronchial epithelial cell line BEP2D induced by α-particle exposure.Methods Glutathione Peroxidase (GPX),Catalase (CAT) and Glutathione (GSH) assay kits were employed to detect GPX and CAT enzyme abilities and the levels of GSH in BEP2D,RH21 ( passage 21 of α-particle-irradiated BEP2D cells),and BERP35T-1 cells (derived from nude mice bearing malignant transformed cells generated from cells of passage 35 of α-particle-irradiated BEP2D cells).MTT assay were used to test the growth rate of BEP2D,RH21 and BERP35T-1 cells treated with 0,30,60,90,120,and 150 μmoL/L H2O2.Colony-forming test and MTT assay were used to examine the cell survival fraction and the growth rate of BEP2D,RH21 and BERP35T-1 cells exposed to 0,2,4,and 8 Gy of γ-rays,respectively.Results GPX and CAT enzyme activities in RH21 and BERP35T-1 cells were obviously lower than in BEP2D( t = 5.75-67.92 ,P ＜ 0.05 ).CAT enzyme activity in BERP35T-1 was lower than that in RH21 cells (t =22.25 ,P ＜0.01 ).Compared to BEP2D cells,decreased level of GSH was detected in BERP35T-1 cells(t = 7.76,P ＜ 0.05 ),but there was no change in RH21.After treatment with 30,60,90,120,and 150 μmol/L H2O2,the growth rates of BEP2D were all higher than those of BERP35T-1 cells(t = 10.37-58.36,P ＜0.01 ).Meanwhile,the growth rates of BEP2D were all higher than those of RH21 cells after treatment with 60,90,120,and 150 μ mol/L H2O2 (t = 29.90-84.68,P ＜ 0.01 ).In addition,compared to BEP2D cells,decreased cell survival fraction and growth rate of BERP35T-1 cells were observed after irradiation with 2,4,and 8 Gy of y-rays ( t = 5.87-34.17,P ＜ 0.05 ).The cell survival fraction and growth rate of RH21 were all lower than those of BEP2D cells at 4 and 8 Gy post-irradition( t =6.33- 45.00,P ＜ 0.05 ).Conclusion The function of antioxidant system decreased in the α-particleinduced transformed cells,which could contribute to the acceleration of cellular malignant transforming process and radiosensitivity.

OBJECTIVETo investigate whether the deletion of PTEN affects the expression of Cu/Zn SOD and the related biology.

METHODSProtein and mRNA expression levels of PTEN, P-Akt, Cu/Zn SOD in the control immortalized wild type mouse embryonic fibroblast cells (PTEN+/+) and PTEN-null cells (PTEN-/-) were evaluated by Western blot and Northern blot respectively. The level of superoxide anions were detected using fluorescent probes. The DNA damage was documented by single cell alkalescence gel assay. MTT was used to study the effect of H2O2 on the proliferation of cells.

RESULTSThe expression of Cu/Zn SOD was down regulated at both protein and mRNA levels, and the level of superoxide anions increased in the PTEN-null cells (PTEN-/-). The phosphorylation level of Akt kinase was up-regulated and the antiproliferative effect of H2O2 decreased in PTEN-/- cells. Furthermore, DNA damage was observed significantly severer in both the blank control and H2O2 treated groups than that in the PTEN+/+ cells.

CONCLUSIONSDeletion of PTEN affects the expression of Cu/Zn SOD. As a result, reactive oxygen species (ROS) keep at a high level, along with decrease of accumulated oxidative damage and the antiproliferative effect of ROS.

Animals ; Copper ; chemistry ; Down-Regulation ; Fibroblasts ; pathology ; Gene Expression Regulation ; genetics ; Hydrogen Peroxide ; metabolism ; Mice ; Mice, Knockout ; PTEN Phosphohydrolase ; deficiency ; genetics ; Phosphorylation ; Reactive Oxygen Species ; Sequence Deletion ; Superoxide Dismutase ; genetics ; metabolism ; Zinc ; chemistry

Objective To study the dose-effect relationship and time-effect relationship of T cell receptor (TCR) gene mutation induced by γ-rays in lymphocytes of human peripheral blood.Methods Samples of peripheral blood were collected from 10 healthy adults and lymphocytes were separated.Four samples from males used to fit time-effect curve were exposed to γ-rays at the doses of 0,0.5,1.0,1.5,2.0,2.5,3.0,3.5,4.0,and 5.0 Gy,respectively,and 6 samples from 3 males and 3 females used to fit dose-effect curves were exposed to γ-rays of the dose of 2 Gy.Flow cytometry was used to detect the mutation frequency of TCR gene (TCR MF).Radiation dose-effect curves and time-effect curves were fitted and optimal mathematical models were selected respectively.Results The optimal mathematical model for radiation dose-effect was quadratic equation model:TCR MF = 92.14 + 22.61D2 (R2adj = 0.65).The optimal mathematical model for radiation time-effect was quadratic polynomial equation model:TCR MF = 3.74 + 743.66T + 308.64T2 (R2adj = 0.79).Conclusions TCR MF is increased as the γ-rayirradiation dose increases within the range of 0-5 Gy,and TCR MF is increased with the lapse of time within the range of 4 days after γ-ray radiation.

Objective To observe the effect and the mechanisms of ferulic acid on radiationinduced damage of mice peripheral blood and bone marrow hematopoietic function.Methods Ninety-six mice were randomly divided into sham irradiation group,irradiation group,positive drug group and 10,30,90 mg·kg-1 ·d-1 ferulic acid group,16 mice per group.Mice were exposed to 3.5 Gy γ-rays 24 h after first drug taken.Then,mice were given drugs for 7 d after irradiation.White blood cells in peripheral blood of 10 mice per group were counted 2 d before irradiation and 3,7,10,15 and 22 days after irradiation.The bone narrow of the other six mice was taken to detect the micronuclei frequency of polychromatic erythrocyte,the hematopoietic progenitor cell colony formation capacity,Thbd and HMGB1 protein expressions in mice bone marrow on the seventh day after irradiation.Results Compared with the irradiation alone group,the treatment of mice with ferulic acid 90 mg· kg-1 · d-1 increased the number of white blood cells in peripheral blood at 3,10,15 and 22 d after irradiation (t =2.267,2.399,1.945,2.828,P ＜ 0.05).Treatment with mice with ferulic acid 90 mg· kg-1 · d 1 decreased the micronuclei rate of erythrocytes in irradiated bone marrow (t =4.013,P ＜ 0.05),increased the clone numbers of CFU-E,BFU-E and CFU-GM of hematopoietic progenitor cells (t =2.366,2.953,3.115,P ＜0.05),improved the relative expression of the Thbd protein in bone marrow and the HMGB1 protein in nuclear (t =17.75,23.39,P ＜ 0.01).Conclusions Ferulilc acid could protect the bone marrow hematopoietic of mice exposed to irradiation by regulating the expressions of Thbd and HMGB1 protein,and then accelerate the peripheral cells recovery.

OBJECTIVETo study the chemical constituents of an active fraction (DSS-A-N-30) from Danggui Shaoyao San.

METHODDSS-A-N30 was prepared by macroporous resin chromatography, the compound was isolated by column chromatography on silica gel and RPC-18, the structure was elucidated by spectroscopic methods.

RESULTA new monoterpene glycoside was isolated and identified from DSS-A-N-30.

CONCLUSIONThe new monoterpene glycoside was identified as 4"-hydroxyl-albiflorin.

Bridged-Ring Compounds ; analysis ; isolation & purification ; Chromatography, Gel ; Drugs, Chinese Herbal ; chemistry ; Magnetic Resonance Spectroscopy ; Monoterpenes ; analysis ; isolation & purification

OBJECTIVETo explore the operative method for the treatment of syndesmosis injury in ankle fractures.

METHODSA retrospective study was done on 21 ankles of 20 patients included male 11 and female 9;the range of age were from 27 to 52 years with an average of 36 years) with syndesmosis injury in closed ankle fractures from September 2005 to December 2007. All patients with ankle fractures and syndesmosis injury were diagnosed by the history, physical examination and radiology, then treated with open reduction, internal fixation, and syndesmotic stabilization with a three-cortices syndesmotic screw according to the Lauge-Hansen classification system. Radiological evaluation comprised tibiofibular overlap, total clear space and medial clear space. The clinical effects were evaluated according to modified Baird-Jackson standard.

RESULTSAll patients were followed up from 1.0 to 2.2 years with an average of 1.3 years. Radiographic measurements were detailed as follows: tibiofibular overlap averaged (0.46 +/- 3.56) mm in preoperative and (7.14 +/- 0.62) mm in postoperative; mean total clear space (5.69 +/-0.88) mm in preoperative and (3.28 +/- 0.39) mm in postoperative; medial clear space averaged (5.67 +/- 1.23) mm in preoperative and (3.12 +/- 0.33) mm in postoperative; tibiofibular overlap in mortise view averaged (-0.87 +/- 0.96) mm in preoperative and (2.91 +/- 0.30) mm in postoperative. There was significant difference above data between preoperative and postoperative (P < 0.01). Four cases were confirmed minor tibiofibular diastasis through CT scans during postoperative. The modified Baird-Jackson scoring was from 62 to 98 scores with an average of (86.24 +/- 13.26) score at the final review. Of them, 13 ankles had not pain; 16 ankles reported no instability complaints; 11 ankles gained normal walking ability; 8 ankles could run normally; 11 ankles could return work without any restrictions. Activity of ankle in dorsiflexion, plantar flexion, inversion and eversion were respectively (21.05 +/- 5.00) degrees, (33.57 +/- 5.76) degrees, (19.48 +/- 4.57) degrees and (24.05 +/- 4.86) degrees. Three cases had radiological and clinical manifestations of osteoarthritis, but no breakage of syndesmotic screw in all cases. There were excellent results in 12 cases, good in 2, fair in 4, poor in 3.

CONCLUSIONThe treatment for the syndesmosis diastasis with a three-cortices screw fixation in ankle fractures is effective. Good functional outcome can be obtained with anatomical restoration of the tibiofibular syndesmosis. The repair of deltoid ligament is important for stability of the lower tibiofibular syndesmosis. Removal of the screw before weight loading should be performed to avoid possible screw breakage.

Adult ; Bone Screws ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; instrumentation ; Fractures, Bone ; diagnostic imaging ; physiopathology ; surgery ; therapy ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Tarsal Bones ; diagnostic imaging ; injuries ; physiopathology ; surgery ; Tomography, X-Ray Computed

OBJECTIVETo explore the clinical results of arthroscopy-assisted open reduction and internal fixation for the treatment of acute Lauge-Hansen stage IV ankle fractures.

METHODSA prospective cohort of study was done in 42 patients of acute ankle fractures treated with arthroscopy-assisted open reduction and internal fixation from January 2008 to December 2009. According to Lauge-Hansen classification, there were 26 supination-eversion stage IV fractures, including 11 females and 15 males; with an average age of (36.8 +/- 11.7) years; and there were 16 pronation-eversion stage IV fractures, including 6 females and 10 males, with an average age of (37.6 +/- 11.2) years. All the patients were treated with arthroscopy-assisted open reduction and internal fixation,and intra-articular disorders were observed. Syndesmotic diastasis was treated with three-cortices syndesmotic screw, and osteochondral injuries were disposed by Cheng-Ferkel staging system. The clinical effects were evaluated according to AOFAS scores.

RESULTSAll the patients were followed up for 1 year. At the final review,the mean AOFAS score was (92.00 +/- 9.32). Twenty-eight patients got an excellent result, 11 good, 3 fair. Lesions of the cartilage were found in 31 cases. The AOFAS scores of patients with osteochondral lesions were higher than those of patients without osteochondral lesions. The AOFAS scores of patients with osteochondral lesions below C grade were higher than those of patients with osteochondral lesions above C grade. The AOFAS scores of patients with osteochondral lesions were higher than those of patients with osteochondral lesions above C grade. The AOFAS scores of patients with syndesmosis fixation were lower than those of patients without syndesmosis fixation.

CONCLUSIONIntra-articular disorders are common in Lauge-Hansen stage IV ankle fractures, and osteochondral lesion and sydesmosis diastasis are the most important prognostic factor that leads to unsatisfactory ankle functions. Arthroscopy-assisted open reduction and internal fixation for the treatment of acute ankle fractures is not only helpful to anatomical reduction of the articular surface, but also can improve the clinical results with effective diagnosis and treatment of combined intra-articular disorders.

Adolescent ; Adult ; Aged ; Ankle Joint ; surgery ; Arthroscopy ; methods ; Female ; Fracture Fixation, Internal ; methods ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Young Adult

OBJECTIVETo analyze the characteristics of cervical minimal deviation adenocarcinoma (MDA) and the methods of diagnosis and treatment.

METHODSA retrospective study was carried out to evaluate the clinical and pathological data of 15 patients with MDA treated from 1992 to 2007.

RESULTSThe average age of the 15 patients was 42.3 years. The main symptoms were increased discharge and irregular vaginal bleeding. Preoperative Pap smears showed adenocarcinoma in 3 cases (27.3%). The diagnosis of MDA was confirmed in 8 cases by cervical punch biopsies (53.3%) and 2 cases by conization. Several cysts were noted in sections of the endocervix. Microscopic examination showed glands irregular in size and shape. However, the deviation of tumor cells was minimal. Immunohistochemistry revealed positive expression of CEA and alpha-SMA. The mean follow-up time was 51.0 months. The overall 5-year survival rate was 85.7%. Four cases experienced recurrence in the vagina and pelvis at 2 years after operation. Three cases died of the disease relapse with an average survival time of 36.3 months.

CONCLUSIONCervical minimal deviation adenocarcinoma is rare, with minimal deviation of cell shape from the normal cervical cells and difficult in diagnosis. A deep biopsy or conization is necessary when punch biopsy is not sufficient for diagnosis. Immunohistochemistry is helpful to make an accurate diagnosis. Surgery is the first choice for cervical minimal deviation adenocarcinoma. Radiotherapy and/or chemotherapy should be given if needed. The prognosis can be improved if a proper treatment plan is carried out.

Actins ; metabolism ; Adenocarcinoma ; diagnosis ; pathology ; therapy ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoembryonic Antigen ; metabolism ; Cervix Uteri ; pathology ; Chemotherapy, Adjuvant ; Cisplatin ; administration & dosage ; Conization ; Epirubicin ; administration & dosage ; Female ; Fluorouracil ; administration & dosage ; Follow-Up Studies ; Humans ; Hysterectomy ; methods ; Middle Aged ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Papanicolaou Test ; Radiotherapy, Adjuvant ; Retrospective Studies ; Survival Rate ; Uterine Cervical Neoplasms ; diagnosis ; pathology ; therapy ; Vaginal Smears