Objective To study the effects of antisense oligodeoxynucleotides(ODN) of hEST2 (AODN) on telomerase activity and proliferation in ovarian cancer cell lines SKOV3 and COC1 Methods Antisense and sense human telomerse catalytic sub unit (hEST2) phosphorothioate (SODN)and random ODN were designed, synthesized and transfected into SKOV3 and COC1 cells by lipofectamine The expression of hEST2 mRNA and telomerase activity in SKOV3 and COC1 were tested by reverse transcription polymerase chain reaction and telomeric repeat amplification protocol before and after transfection The proliferation and growth in SKOV3 and COC1 were also investigated by methyl thiazolyl tetrazolium and growth curve before and after transfection Results AODN could down regulate the expression of hEST2 mRNA, inhibit telomerase activity and proliferation of ovarian cell lines The efficiency depends on dose and period of administration At 48 h, 30 ?mol/L AODN had the highest activity The expression of hEST2 mRNA were declined 54 6% and 44 6% in SKOV3 and COC1 respectively And also the inhibition of telomerase activity were 47 9% and 42 7% respectively in the two cell lines Conclusions AODN of hEST2 clearly inhibited the proliferation of ovarian cancer cell lines hEST2 may thus be a new target of gene therapy in ovarian carcinoma

ObjectiveRecurrent rupture of Achilles tendon is a severe complication after primary repair.The optimal treatment remains controversial.This study was in aiming to explore the risk factors of recurrent ruptures of Achilles tendon,to evaluate the indication,outcome and complications of reconstructing Achilles tendon recurrent ruptures by V-Y advancement flap and Flexor Hallucis Longus tendon (FHL) transfer.MethodsWe retrospectively studied 16 cases who underwent reconstruction of Achilles tendon recurrent ruptures from Mar,2006 to Jan,2010.Among them there were 12 males and 4 females with the mean age of 50.9 years(35-72 years).The period of time between recurrent rupture and primary operation was 6-49 weeks(ave.21.8 weeks).The follow-up interval was 6-52 months (ave.27.5 months).During operatin,4 cases with a rupture gap less than 4 cm had received V-Y advancement flap and 12 cases with a rupture gap more than 4 cm had received FHL transfer.At the time of follow-up,all patients were assessed with the American Orthopaedic Foot and Ankle Society(AOFAS) Ankle-Hindfoot Scale and Leppilahti Achilles Tendon Repair score.ResultsThe average AOFAS score had increased from 70.2±8.5 preoperatively to 92.4±6.1postoperatvely.Leppilahti Achilles Tendon Repair score had increased from 74.8±6.2 preoperatively to 91.7±4.8 postoperatvely.MRI of Achilles tendon showed even signal without signs of tear.ConclusionV-Y advancement flap could repair a rupture gap within 4 cm,FHL transfer is suitable for rupture gap over 4 cm in reconstruction of Achilles tendon recurrent ruptures.

Objective To discuss the pathomechanism of medial ankle instability after medial ligament injury. Methods The study involved 15 patients including eight males and seven females, at average age of 40 years (range 22-58 years). There were two patients with acute ankle sprain, three with chronic ankle injury, three with congenital pes planus, five with posterior tibial tendon dysfunction, one with pronation external rotation ankle fracture and one with old avulsion medial malleolus. All patients were treated surgically with deltoid ligament repair, when eight patients further received calcaneal lengthing osteotomy, five received medial cuneiform close wedge osteotomy and one received medial shift calcaneal osteotomy. All patients were followed up for 7-56 months. American Orthopedic Foot and Ankle Society (AOFAS) ankle-hindfoot scale was used for pre- and post-operative evaluation of the function. All data were analyzed using t test. Results The clinical data of one patient was excluded from the statistical analysis because AOFAS ankle-hindfoot scale could not be applied for evaluating the acute ankle fracture. The AOFAS ankle-hindfoot scale was 42.4 + 10.6 before surgery and 89.8 +6.2 at the final followup for 14 patients (P＜0.05). Conclusions Deltoid ligament is an important anatomical structure.More attention should be paid to its repair under following aonditions: ( 1 ) the injury of the deltoid ligament involves tibiospring and tibionavicular ligament; (2) patients have congenital pes planus and posterior tibial tendon dysfunction; (3) patients present with ankle fractures and have history of repeated ankle sprain.

ObjectiveTo evaluate effect of periarticular osteotomy in the treatment of asymmetrical ankle arthritis.MethodsSixty-five patients with asymmetrical ankle arthritis were treated with periarticular osteotomy between February 2005 and May 2011,including 43 females and 22 males,aged from 35 to 74 years (mean,55.5 years).According to the Takakura classification of ankle arthritis,there were 29 patients in grade 2,32 in grade 3 and 4 in grade 4.Supramalleolar tibial osteotomy were performed in 20 patients,supramalleolar tibial and fibular osteotomy in 12,supramalleolar osteotomy combined with calcaneal osteotomy in 30 patients,and supramalleolar tibial and fibular osteotomy combined with calcaneal osteotomy in 3patients.Forty three patients underwent ligament reconstruction procedures.Based on radiographs,the tibial anterior surface angle (TAS) and tibial lateral surface angle (TLS) were compared before and after operation.AOFAS-AH score was used to evaluate the function of the ankle.ResultsFifty-nine patients were followed up for an average of 35.7 months (range,7 to 94 months).Bone healing was observed in all patients,and the average healing time was 7 to 8 weeks.The average AOFAS-AH score was improved from 49.7 points preoperatively to 78.6 points 12 months postoperatively.Tbe average TAS and TLS was improved from 86.1° and 70.0° preoperatively to 93.9° and 81.5° 6 months postoperatively,respectively.Delayed wound healing occurred in 5 patients,which was resovled after nursing care.Forty-two patients felt excellent about results,15 felt good and 2 fell fair.ConclusionPerarticular osteotomy is a sound method in the treatment of asymmetrical ankle arthritis,based on chosing proper patients.The procedure can decrease the contact pressures on the degenerated cartilage and prolong the life span of the ankle.

Objective To investigate the expression and clinical significance of myeloid cell leukemia-1 (MCL-1) and F-box and WD repeat domain-containing 7 (FBW7) in breast cancer polyploid induced by spindle poisons. Methods (1) Nocodazole spindle poison was used to treat breast cancer cell MDA-MB-231. The morphological changes of cells were ob?served under microscope, and cells were harvested in 0, 6, 12, 24, 48 and 72 h. The cell cycle and DNA-ploidy changes were examined by flow cytometry. The expressions of FBW7 and MCL-1 proteins were detected by Western blot assay. (2) A multikinase inhibitor (Sorafenib) with Nocodazole or Taxol was used to treat MDA-MB-231 cells. MCL-1 protein expression was detected by Western blot assay after 48 h treatment. The cell cycle and DNA-ploidy changes were examined by flow cy?tometry after 48 h treatment. MTT method was used to observe cell proliferation after 48 and 72 h treatment. Results (1)Af?ter treatment by Nocodazole, polyploid characteristics of large cell size and nucleus were appeared. The percentages of octa?ploid were (0.8±0.2)%, (8.5±2.3)%, (7.8±2.0)%, (9.9±0.9)%, (28.2±0.8)%and (35.1±4.9)%after 0, 6, 12, 24, 48 and 72 h treatment, showing the increasing trend in turn (P<0.001). The number of polyploidy (tetraploid and octaploid) cells was as high as (97.6±0.7)%after 48 h treatment. The expression level of FBW7 protein was decreased significantly but the expres?sion of MCL-1 protein was increased significantly after 48 h treatment. (2) After 48 h treatment, the expression level of MCL-1 protein, polyploidy percentage and cell proliferation decreased significantly in Nocodazole+Sorafenib group and Taxol+Sorafenib group compared with those of Nocodazole group and Taxol group (P<0.05). Conclusion The lower expression of FBW7 protein and over-expression of MCL-1 protein are correlated with the formation of breast cancer polyploidy. Sorafenib can reduce polyploid tumor cells by inhibiting MCL-1 protein expression.

AIM:To observe the effect of altitude hypoxia on ?-aminobutyric (GABA) content and prepro-somatostatin mRNA (PPS-mRNA) in the rat hypothalamus. METHODS: Using altitude hypoxia model,in situ hybridization and amino acid analyzer, the number of PPS-mRNA and GABA content in rat hypothalamus was determined. RESULTS:After altitude hypoxia, the contents of GABA in hypothalamus and the number of PPS-mRNA neurons in periventricular nucleus (PeVN), paraventricular nucleus (PaVN) and arcuate nucleus (ArcN) increased significiantly. Bicuculline, a GABA receptor antagonits, could enhance PPS-mRNA expression evoked by altitude hypoxia, but had no effect on GABA content. CONCLUSION: Altitude hypoxia can induce neurotransmitter imbalance of hypothalamus.

AIM: To observe the effect of altitude hypoxia on glutamate, aspartate and nitric oxide synthase (NOS) in the rat hypothalamus. METHODS: Using altitude hypoxia model, amino acid analysis system and the NADPH-d histochemistry, we determined the content of glutamate, aspartate and the number of NADPH-d neurons in the rat hypothalamus. RESULTS: After altitude hypoxia, the contents of glutamate, aspartate in the hypothalamus of rats were increased significantly, densely and deeply stained NADPH-d neurons were seen in hypothalamic paraventricular nucleus (PVN)and supraoptic nucleus(SON). If rat were pretreated with the NMDA receptor blockers Ketamine (ip,40 mg/Kg)or AP-V(i.c.v, 10 ?g) , the number of NADPH-d neurons in the rat hypothalamic PVN and SON was markedly less than that in corresponding altitude hypoxia group. CONCLUSION: NMDA receptor may take part in the expression of hypothalamic NOS induced by altitude hypoxia.

Objective To study on the different results of a minimally invasive method to reconstruct the ligaments of the lateral ankle using semitendinosus tendon autograft and tendon allograft.Methods Data of 68 patients with chronic ankle instability who had undergone lateral ligament reconstruction from September 2006 to June 2011 were retrospectively analyzed.In the group of semitendinosus autograft,there were 32 patients (19 males,13 females) with an average age of 32.4 years old.Semitendinosus was harvested through 2 small knee incisions.While in the group of tendon allograft,there were 36 patients (22 males,14 females) with an average age of 34.2 years old.For the ankle reconstruction,4 small incisions of 5 mm each were made at the medial and lateral side of the fibular tip,the talar neck,and the middle of the calcaneus.Anatomical reconstruction of the anterior talofibular ligament and calcaneofibular ligament was then performed through these small incisions.AOFAS questionnaires were used to measure clinical outcomes,donor site morbidity and patients' satisfaction.Preoperative and postoperative stress tests were performed and radiographic parameters were measured.Results The average operation time of the autograft group 85.5 ± 11.5 min was significantly longer than that in the allograft group 58.1±10.2 min,but the fever days in the latter 5.5±1.5 d was significantly longer than in the former 2.5±1.2 d.In autograft group,23 patients got followed up,and the mean period of follow-up was 33.5±6.7 months.The mean AOFAS score increased from 62.3±8.2 to 95.1 ±7.5.In allograft group,26 patients got followed up,and the mean period of follow-up was 28.5±6.7 months.The mean AOFAS score increased from 60.2±8.4 to 94.8±5.5.There were 5 patients (3 of autugraft group and 2 of allograft group) with residual instability on uneven ground.One case of the autologous group had instability in daily life.No patient presented weakness or disability in the donor site.The satisfaction level of the autograft group was excellent in 16,good in 5 and bad in 2.Significant improvement in stress radiographic parameters was noted for the talar tilt angle,with reduction from a mean of 14.0° to 3.8°; anterior talar displacement reduced from a mean of 12.3 to 4.6 mm.The satisfaction level of the allograft group was excellent in 17,good in 5 and bad in 4.The talar tilt angle reduced from 13.0° to 3.6°; anterior talar displacement reduced from 11.5 to 4.3 mm.Conclusion There is no differences in efficacy could we found in using these 2 kinds of materials.The process of healing and rehabilitation of tendon autograft is slightly faster than the tendon allograft,but the allograft tendon has many advantages such as limited injury,simple procedure andavoiding of donor site morbidity.

Objective To evaluate the curative effect and related factors of the treatment for the osteochondral lesion of talus (OLT) by the osteochondral autografts harvested from medial femoral condyle of the ipsilateral knee.Methods The data of 15 OLT patients was retrospectively analyzed who received the operation during July 2009 to November 2012.There were 8 males,and 7 females,with an average age of 49.6±17.2 years (range,19-73 years).International Knee Documentation Committee (IKDC) and Lysholm score were used to evaluate the knee function preoperatively and postoperatively respectively.The ankle functions and pain were assessed according to American Orthopaedic Foot and Ankle Society (AOFAS) ankle and hind-foot score and visual analogue scale (VAS) preoperatively and postoperatively respectively.Correlations between age or follow-up duration and all the score increments were further analyzed by Spearman rank correlation test.Results Twelve patients complete the follow up,with an average duration of 21.8±10.4 months.The average IKDC,Lysholm,AOFAS,and VAS were 90.91±6.44,95.33±8.00,63.58±18.50,and 7.25±1.54 respectively before operation,and 85.63±11.89,90.75±11.83,90.33±4.98,2.17±1.19 respectively after operation.Correlation coefficients of age between AOFAS,Lysholm,and IKDC score increments were -0.74,-0.63,and-0.76,respectively.There were 4 cases which received excellent effect (33％),5 cases (42％) good and 3 (25％) fair.Conclusion The ankle joint function of patients with OLT recovered well by treated with osteochondral autografts harvested from medial femoral condyle of the ipsilateral knee operation.A correlation was found between patients' age and postoperative functional recovery of ankle and knee joint.