Objective To clarify the role of claudin-4 in endometrial tumorigenesis and explore claudin-4 be as potentially useful agent in the treatment of endometrial carcinoma.Methods The expression of claudin-4 in 62 endonetrioid endometrial carcinoma (EEC),30 atypical hyperplasia endometrial tissue and 60 human normal endometrium was determined using immunohistochemistry and realtime PCR.Ninety female BALB/c mice were transplanted with Ishikawa endometrial cancer cells,which were divided into three groups with different intraperitoneal treatments with cisplatin,paclitaxel and saline solution.After the observation period,the tumors were extracted and stained with monoclonal antibody against claudin-4.The messenger RNA expression of claudin-4 was also detected using real-time PCR.Results Among the EEC samples,34％ (21/62) showed medium staining for claudin-4 and 66％ (41/62) showed intense staining.In atypical hyperplasia group,27％ (8/30) showed weak staining,53％ (16/30) showed medium staining and 20％ (6/30) showed intense staining for claudin-4.Of the normal endometrial tissue,47％ (28/60) showed weak staining and 53％ (32/60) showed no staining for claudin-4.According to real-time PCR,the relative quantity of claudin-4 was 170 ± 12 in EEC group,89 ± 15 in atypical hyperplasia group and 18 ± 3 in normal endometrium.Compared with those in atypical hyperplasia group and normal endometrium group,the protein and mRNA expression of claudin-4 were significantly increased in the group of EEC (all P ＜ 0.05).In the study of Ishikawa xenografts,no significant changes in tumor volume and claudin-4 expression were shown in paclitaxel group compared with that in the control group.Nevertheless,a significant reduction of the tumor growth and a significant decrease in claudin-4 expression were observed in cisplatin group.After cisplatin treatment,the tumor volume was significantly decreased [(0.51 ±0.21) versus (0.73 ±0.12) cm3],and the mRNA expression of claudin-4 was also significantly decreased (153 ± 35 versus 273 ± 27).Conclusion These results demonstrate that claudin-4 is strongly expressed in EEC,which may be a useful biomarker to monitor the effects of chemotherapy in patients with endometrial carcinoma.

Objective To construct a cell line with long time secreting endostatin. Methods Human endostatin cDNA containing interleukin-2 (IL-2) secreting peptide was cloned into eukaryotic expression plasmid pSNA2.0 to construct recombinant plasmid pSNA2.0/Endostatin. The plasmid pSNA2.0/Endostatin was transfected into HEK293 cells by cationic liposome. The positive cell clones were selected by G418 and named hED/293. The expression of endostatin protein was analyzed by Western-blot. Results Endostatin could be determined in the supernatant of hED/293 cells. Conclusion The recombinant eukaryotic expression vector is correctly constructed. The human endostatin protein can be expressed and secreted.

Objective To establish the model of glioma by hypodermic of C6 cells line into the nude mice and observe the growth characteristics. Methods The C6 gliocyte suspension was injected into the left subaxillary hypoderma of nude mice. The diameter and the volume of the tumor were measured and calculated, and the volume-time growth curve was plotted. The cells of tumor were observed under the HE and glial fibrillary acidic protein (GFAP) immono-histochemistry staining. Results The tumor appeared in inoculation area 7 d after injection, and presented nearly exponential development after about 20 d. The survival time of the nude mice is 22～48 d. The tumors were with sharp border, affluent blood vessel, and a great number of GFAP-positive cells. Conclusion The model of glioma can be well induced by hypodermic of the cultured C6 gliocyte into nude mouse.

OBJECTIVETo explore methods of treating middle and distal tibia nonunion with the treatment of advanced bone graft combined with locking compression plate.

METHODSFrom January 2011 to December 2012, 12 patients with middle and distal tibia nonunion were treated with advanced bone graft combined with locking compression plate. Among patients, there were 8 males and 4 females aged from 20 to 69 with an average of 47 years old. The time from first injuries to bone nonunion was from 9 months to 5 years, avergaed 19 months. Four cases were treated with external fixation, 6 cases were treated with plate fixation, 2 cases of 12 patients occurred broken of plate and nail. Eleven patients were non-infective bone nonunion and 1 patient was infective bone nonunion. Preoperative X-ray and CT showed all patients had sequestration and formation of ossified bone with different degrees. Operative time, blood loss, wound healing were observed, fracture healing time was evaluated by postoperative X-ray. Johner-Wruhs scoring standards was used to evaluate ankle joint function after operation at 10 months.

RESULTSOperative time ranged from 90 to 185 min with an average of (125.00±20.15) min; blood loss ranged from 225 to 750 ml with an average of (415.00±120.00) ml. All patients were followed up from 10 months to 2.5 years with an average of 1.5 years. Postoperative X-ray showed bone union was formed around fracture after operation at 4 months in all patients, 3 cases obtained bone healing within 6 months after operation, 9 cases obtained from 8 to 12 months. No infection, injury of nerve and vessles, and broken of plate and nail were ocurred. According to Johner-Wruhs scoring at 10 months after operation, 10 cases obtained excellent results, 1 good and 1 moderate.

CONCLUSIONAdvanced bone graft combined with locking compression plate, which can build fracture multi-point supporting based on full compression of bone nonunion to get effective fixation, is an effective method in treating middle and distal tibia nonunion.

Adult ; Aged ; Bone Plates ; Bone Transplantation ; Female ; Fracture Healing ; Fractures, Ununited ; surgery ; Humans ; Male ; Middle Aged ; Tibial Fractures ; surgery

Objective To observe the effect of APA-BCC(alginate-polylysine-alginate microencapsulated bovine adrenal medullary chromaffin cells)microcapsules transplantation into the subarachnoid space of cancer pain patients on endogenous opioid peptides and catecholamine concentration in cerebrospinal fluid(CSF).Methods The different doses of APA-BCC microcapsules were transplanted into the spinal subarachnoid space of cancer patients with moderate or serious pain.The concentrations of Leu-enkephalin(L-EK),β-endorphin(β-EP),dynorphin A(DynA),noradrenaline(NA)and adrenaline(AD)in CSF were tested.Results L-EK concentration in CSF increased remarkably after transplantation,and the increase was most remarkable when transplantation doses were 1.0×107and 1.25×107;there were no remarkable changes of β-EP,DynA,NA and AD after transplantation.Conclusion The analgetic effects of APA-BCC microcapsules tranplantation may correlate with the increase of L-EK in CSF of cancer pain patients;the dose of 1.0×107 and 1.25×107cells may be the most effective dose.

OBJECTIVETo explore morphological character and clinical significance of superior-posterior acetabular wall by anatomically measuring and quantitatively analyzing thickness of posterior acetabular wall, then provide a theoretical reference for clinical treatment of acetabular fracture.

METHODSFifteen adult formalin-preserved cadaveric pelvises (8 males and 7 females) were used for this investigation. Excess soft tissue was removed and the whole acetabular posterior walls were marked with "angle" sector method and the thickness was measured with caliper in different levels of the different split points. The measurement results were validated and analyzed statistically.

RESULTSAt 5 mm away from acetabular rim, the average thickness of superior-posterior acetablar wall fluctuated between (6.47±0.61) mm and (7.43±0.71) mm; the average thickness of inferior-posterior acetabuluar wall fluctuated between (5.62±0.51) mm and (6.33±0.61) mm; the average thickness of acetabular roof fluctuated between (7.71±0.74) mm and (8.27±0.99) mm. There was no statistical difference between average thickness of superior-posterior wall of acetabulum and inferior-posterior wall of acetabulum (P>0.05), but the average thickness of acetabular roof was significantly larger than superior-posterior acetabular wall (P<0.05). At 10 mm away from the acetabular rim, the average thickness of superior-posterior acetabular wall fluctuated between (8.81±0.67) mm and (13.35±0.89)mm; the average thickness of inferior-posterior acetabular wall fluctuated between (7.02±0.63) mm and (7.66±0.69) mm; the average thickness of acetabular roof fluctuated between (14.46±0.97) mm and (17.05±1.35) mm. Comparatively, the average thickness of superior-posterior acetabular wall was significantly larger than inferior-posterior wall of acetabulum (P<0.05), and the average thickness of acetabular roof was significantly larger than superior-posterior acetabular wall (P<0.01). At 15 mm away from the acetabular rim, the average thickness of superior-posterior acetabular wall fluctuated between (12.08±0.78) mm and (19.84±1.03) mm; the average thickness of inferior-posterior acetabular wall fluctuated between (10.17±0.76) mm and (11.12± 0.77) mm; the average thickness of acetabular roof fluctuated between (23.23±1.12) mm and (26.01±1.53) mm. Comparatively, the average thickness of superior-posterior wall of acetabulum was significantly larger than inferior-posterior acetabular wall (P<0.01), and the average thickness of acetabular roof was significantly larger than superior-posterior acetabular wall (P< 0.01).

CONCLUSIONThe thickness of entire acetabular posterior edge revealed an increasing tendency from inferior-posterior wall to the superior-posterior wall to acetabular roof. And this trend became more obvious with increasing distance away from acetabular rim. Therefore, the superior-posterior acetabular wall could not only maintain the stability of hip joint but also bear loading.

Acetabulum ; anatomy & histology ; injuries ; surgery ; Female ; Humans ; Male

OBJECTIVETo explore the clinical symptom and effect of arthroscopic treatment of symptomatic anterior cruciate ligament (ACL) cysts of the knee.

METHODSClinical data from 12 symptomatic ACL cysts patients from January 2005 to December 2010 were retrospectively analyzed,including 8 males and 4 females,with an average age of (33.7±9.5) years old (ranged, 19 to 53 years old). The locations were the left knee in 5 cases and the right knee in 7 cases. The disease duration ranged from 3 to 48 months,with a mean of (15.8±13.2) months. All cysts were arthroscopically resected. Range of motion was measured preoperatively and postoperatively, and Lysholm scoring system was used to evaluate the knee function.

RESULTSAll the incisions healed by first intention, and no complications occurred. Twelve patients were followed up for an average of (32.3±6.6) months(ranged, 24 to 48 months). The symptoms of arthralgia,swelling and interlocking of the affected knees disappeared. There was no recurrence during the follow-up. There were significant differences in the range of motion and Lysholm score between pre-operation and post-operation.

CONCLUSIONArthroscopic surgery, showing its advantages of minimal invasion and rapid recovery,is an effective measure in the treatment of ACL cysts.

Adult ; Anterior Cruciate Ligament ; physiopathology ; surgery ; Arthroscopy ; methods ; Cysts ; physiopathology ; surgery ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Range of Motion, Articular

Objective: To observe the time-effect relationship of moxibustion for primary dysmenorrhea (PD) due to stagnation and congelation of cold-damp, thus explore the optimal choice of moxibustion duration, and provide evidence for achieving satisfactory efficacy in moxibustion treatment. Methods: A total of 90 patients with PD due to stagnatin and congelation of cold-damp were divided into three groups by the random number table method, with 30 cases in each group. All the patients in the three groups were given moxibustion treatment at Guanyuan (CV 4), 20 min in group A, 40 min in group B and 60 min in group C. The changes in the pain measurement score in the three groups were observed after treatment. Results: After treatment, there were significant differences in the clinical efficacy among the three groups (P<0.05); the clinical efficacy was better in group B and group C than that in group A (P<0.05), and that in group B was better than that in group C (P<0.05). Besides, the pain measurement score changed significantly after treatment in the three groups (all P<0.05), and the between-group differences were also statistically significant (P<0.05); the pain measurement scores in group B and group C were lower than that in group A (P<0.05), and that in group B was lower than that in group C (P<0.05). Conclusion: Given the same stimulating frequency and intervention time of moxibustion, 40-minute duration demonstrates relatively better efficacy for PD due to stagnation and congelation of cold-damp.

This study was purposed to investigate the effect of phycocyanin at different concentration on proliferation of K562 cells, to detect the changes of integrin beta1 expression and intracellular focal adhesion kinase (FAK) gene expression on the surface K562 cells treated with phycocyanin, and to explore the possible mechanism of integrin beta1 effect on phycocyanin inhibiting proliferation of K562 cells. The expression level of integrin beta1 on the surface of K562 cells was evaluated by flow cytometry (FCM); the growth of K562 cells treated with phycocyanin was measured by MTT assay; the expression level of FAK mRNA was analyzed by relatively quantitative RT-PCR after four-day culture of K562 cells with phycocyanin of 40 microg/ml, 80 microg/ml and 160 microg/ml, respectively. The results showed that integrin beta1 expression on the surface of K562 cells was significantly higher than that in bone marrow mononuclear cells (BMMNC) from normal subjects. Phycocyanin could not change the level of integrin beta1 expression. Phycocyanin could increase the expression of FAK gene on K562 cells and inhibit the proliferation of K562 cells. It is concluded that phycocyanin can inhibit the proliferation of K562 cells through enhancing the conjunction of cell stroma with integrin beta1 on K562 cell surface, up-regulating the expression level of FAK gene in K562 cells, restoring the signaling pathway of proliferation inhibition mediated by integrin beta1. The possible mechanism of phycocyanin in the proliferation inhibition of K562 cells is to increase the expression of FAK gene. The phycocyanin may be considered as a potential agent for inhibition of cancer cell proliferation.
Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Proliferation ; drug effects ; Focal Adhesion Kinase 1 ; biosynthesis ; genetics ; Humans ; Integrin beta1 ; biosynthesis ; genetics ; K562 Cells ; Phycocyanin ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics

BACKGROUNDSnake venom contains a number of components with different pharmacological and biological activities, especially in cancer therapy, and has increasingly become a research focus. This study was designed to isolate and purify a novel anti-clotting protein component from the venom of Agkistrodon acutus, and to explore its physico-chemical properties and biological activity.

METHODSThe venom of Agkistrodon was isolated and purified by ion-exchange chromatography on diethylaminoethyl (DEAE)-Sepharose Fast Flow, molecular sieve filtration through Sephadex G75, SP-Sepharose Fast Flow and molecular sieve filtration through Sephadex G50. We detected the activated partial thromboplastin time (APTT) of the eluant to select the anti-clotting protein component of interest. The molecular weight was determined by sodium dodecyl sulfate-polyacrylamid gel electrphoresis (SDS-PAGE) and liquid chromatography. Its protein content was detected by bicinchoninic acid (BCA).

RESULTSSDS-PAGE vertical gel electrophoresis showed that the anticoagulant factor is a tripolymer composed of three proteins whose molecular weights are 25 KDa, 30 KDa and 50 KDa. The factor contains about 65% percent protein.

CONCLUSIONSA novel anti-clotting protein component was purified by ion-exchange chromatography and molecular sieve filtration from the venom of Agkistrodon acutus and was found to be composed of three kinds of proteins.

Agkistrodon ; metabolism ; Animals ; Anticoagulants ; chemistry ; isolation & purification ; Chromatography, High Pressure Liquid ; Chromatography, Ion Exchange ; Crotalid Venoms ; chemistry ; Electrophoresis, Polyacrylamide Gel ; Proteins ; chemistry ; isolation & purification