The voltage-gated potassium channels Kvl.5 is widely expressed in the plasma membranes of numerous tumor cells and it can contribute to a variety of cellular functions such as proliferation,ap optosis,and cell cycle.Several types of antineoplastic drugs can change the expression of Kv1.5 and then affect the biological processes.Kv1.5 is identified as a novel target for therapy in human cancer.The researches of Kv1.5 will contribute to gain a further understanding of the molecular mechanism of tumor and provide therapeutic opportunity for the prevention and treatment of cancer.

Objective To investigate the inhibition of Hedyotic diffusa injection on the proliferation of hu-man osteosarcoma cell line MG -63 in vitro.Methods The human osteosarcoma cell line MG -63 was subcultivated aseptically in vitro.Different concentration of Hedyotic diffusa injection (50μL/mL,100μL/mL,200μL/mL,400μL/mL) successively acted on such cell .A total three time points were selected to determine the activity and numbers of cells by MTT assay including 12h,24h and 48h.Results The cellular proliferation inhibition rates of human osteosarcoma cell line MG-63 in drug groups of 50μL/mL concentration holes were (2.87 ±2.22)%,(13.42 ±2.14)% and (30.80 ±3.67)%after 12 h,24 h and 48 h.The rates of 100μL/mL concentration holes were (22.25 ±1.58)%, (43.34 ±2.84)%and (66.46 ±2.64)%,after 12 h,24 h and 48 h.The rates gradually increased and had statis-tical meaning,t12h =12.319,t24h =14.573,t48h =12.319,P<0.05;the cell in drug groups of 200μL/ml and 400μL/ml concentration holes was totally dead , and pathological findings showed that there were circular and floating cells in which nucleoplasmic ratio decreased and small fragments of cells increased .Conclusion Hedyotic diffusa extract has a certain inhibition in vitro on the proliferation of human osteosarcoma cell line MG -63.Moreover,the strength of its inhibition is relevant probably with drug concentration ,which deserves a further research .

Objective To detect the expression of HERG (human ether-à-go-go-related gene ) potassium channel in human osteosarcoma,and explore the effects of silencing HERG by small interfering RNA (siRNA)on the proliferation and apoptosis of osteosarcoma cells and the mechanisms responsible for HERG regulation.Methods The expressions of HERG in osteosarcoma MG-63 cells and tissues were detected by reverse transcription polymerase chain reaction (RT-PCR),Western blotting and immunohistochemistry.Next, osteosarcoma cells were divided into three groups:HERG-siRNA group,control-siRNA group and blank group. CCK-8,colony formation,flow cytometry and Tunel assay were used to measure the proliferation and apoptosis of the osteosarcoma cells.Finally,Western blotting analysis was performed to detect the expression of nuclear factor-κB (NF-κB)pathway in osteosarcoma cells treated with HERG siRNA.Results Osteosarcoma cells and tissues were found to highly express HERG.Inhibition of HERG in the osteosarcoma cells significantly inhibited the cell proliferation and induced cell apoptosis.Compared to control-siRNA group or blank group,HERG-siRNA could inhibit the proliferation of MG-63 cells significantly [HERG-siRNA group:(75.34 ± 4.45)%;compared to control-siRNA group:(100.60 ±5.31)%;t =3.64,P =0.007;compared to blank group:(100.00 ±5.66)%;t =3.43,P =0.009].The similar results were obtained from colony formation assay (HERG-siRNA group:134.30 ±11.82;compared to control-siRNA group:225.30 ±11.56;t =5.51, P =0.002;compared to blank group:232.80 ±12.21;t =5.80,P =0.001).HERG-siRNA transfected MG-63 cells demonstrated a significant increase of apoptotic rate compared to control-siRNA transfected cells or untreated cells [HERG-siRNA group:(28.10 ±2.21 )%;compared to control-siRNA group:(9.36 ± 2.42)%;t =5.72,P =0.005;compared to blank group:(10.92 ±2.51)%;t =5.14,P =0.007].This resultwas further confirmed by Tunel assay.The cells transfected with HERG-siRNA (31.57 ±2.08)% dem-onstrated extensive apoptosis,compared with the control-siRNA group [(10.35 ±1.82)%;t =7.69,P =0.002)]or blank group [(7.96 ±0.88)%;t =10.48,P =0.001].Silencing HERG gene down-regulated the cIAP-1,XIAP,Bcl-2,Survivin,P-IκBαand NF-κB p65 expression,compared to the control groups. Conclusion HERG is highly expressed in osteosarcoma.HERG silencing can suppress osteosarcoma progres-sion through NF-κB pathway and suggest that HERG may be a novel molecular target for osteosarcoma therapy and diagnosis.

Objective To investigate the clinical efficacy of our self-designed adjustable weight-bearing brace for AO type B tibial shaft fractures managed by interlocking intramedullary nail.Methods A total of 68 consecutive patients with AO type 42-B tibial shaft fracture who had been managed from April 2013 to March 2015 hy interlocking intramedul]ary nail were recruited into our study.They were randomized into 2 equal groups (n =34).Group A received conventional therapy after operation while group B received auxiliary mauagement with our self-designed adjustable weight-bearing brace after conventional postoperative therapy for one week.The 2 groups were compared at postoperative 1,3 and 6 months and at the final follow-up in terms of visual analogue scale (VAS),weight-bearing status of the affected limb,time for fracture union,Radiographic Union Score for Tibial Fractures (RUST) and Johner-Wruhs scale.Results Of this series,62 cases were followed up for 12 to 18 months (average,14.7 months),5 ones were lost to the follow-up and one withdrew.The mean VAS scores at 3-month and 6-month follow-ups for group B were 2.5 ± 0.8 and 0.9 ± 0.6 respectively,significantly lower than those for group A (3.0 ± 0.9 and 1.4 ± 0.8 respectively) (P ＜ 0.05).In group A at 1-month,3-month and 6-month follow-ups,the weight-bearing status was 44.1％ ± 17.5％,72.0％ ±17.4％ and 86.4％ ±12.5％ while the mean RUST scores were 5.4±1.4,8.7±1.1 and 10.3 ± 1.1,respectively.In group B at 1-month,3-month and 6-month follow-ups,the weight-bearing status was 53.8％ ± 11.0％,84.1％ ± 12.2％ and 94.4％ ± 10.6％ while the mean RUST scores were 6.5 ± 0.8,9.9 ± 0.9 and 11.3 ± 0.8,respectively.There were significant differences between the 2 groups in the above indexes (all P ＜ 0.05).Group B achieved clinical fracture union after an average of 3.3 ±0.7 months,significantly faster than group A (3.9 ± 1.0 months) (P ＜ 0.05).According to the Johner-Wruhs scoring,group A had 19 excellent cases and 12 good ones while group B had 27 excellent ones and 4 good ones,showing a significant difference between the 2 groups (P ＜ 0.05).Conclusions Early application of our self-designed adjustable weight-bearing brace for patients with AO type B tibial shaft fracture managed by interlocking intramedullary nail can reduce postoperative pain,accelerate callus growth,shorten bony healing time and achieve satisfactory functional recovery.

Objective To detect the influence of miRNA-34a (miR-34a) on the proliferation of osteosarcoma and the mechanisms responsible for miR-34a regulation.Methods The osteoblastic cell line MG-63 and Saos-2,human osteoblastic cell line hFOB 1.19,10 osteosarcoma tissues and 10 normal bone tissues were selected.The expression of miRNA-34a in osteosarcoma cells and tissues was detected by quantitative real-time polymerase chain reaction (qPCR).Next,a eukaryotic expression vector named pcDNA/miR-34a was constructed.Then,osteosarcoma cells were transfected with this eukaryotic expression vector and the effects of miR-34a overexpression on the proliferation and growth of osteosarcoma were measured using CCK-8,colony formation and xenograft model of nude mice.Finally,Western blot analysis was used to detect the expression of ether-à-go-go 1 (Eag1) gene in osteosarcoma cells after transfected with pcDNA/miR-34a or a miR-34a inhibitor miR-34a-2'-O-Methyl antisense oligoribonucleotide (miR-34a-2'-O-Me).Results Compared with normal bone tissues and osteoblastic cell line,miR-34a was down-regulated in osteosarcoma cell lines and tissues.Compared with the blank group and the control group,the cell survival rates of miR-34a group of the two cell lines were significantly lower [MG-63 72 h:blank group (40.05±4.82) ％,control group (36.88± 4.66) ％,miRNA-34a group (26.24±6.22) ％;MG-63 96 h:blank group (83.55±5.95) ％,control group (80.13± 4.48) ％,miRNA-34a group (30.21±7.26) ％;Saos-2 72 h:blank group (46.45±8.15) ％,control group (43.33± 6.89) ％,miRNA-34a group (26.81±3.17) ％;Saos-2 96 h:blank group (84.79±4.10) ％,control group (80.14± 3.11) ％,miRNA-34a group (31.77±5.17) ％].The similar results were obtained from colony formation assay (MG-63:blank group 83.40±3.29,control group 80.00±3.06,miR-34a group 24.40±2.71;Saos-2:blank group 85.00±3.32,control group 80.60±3.29,miR-34a group 30.40±4.94).The tumor volumes of osteosarcoma xenograft in the miR-34a group was significantly smaller than that in the blank group and control group after 21 days treatment (all P ＜ 0.001).Overexpression of miR-34a could decrease Eag1 expression in osteosarcoma cell lines while inhibition of miR-34a induced the of expression Eag1 (P ＜ 0.001).Conclusion MiR-34a plays a tumor suppressor role in osteosarcoma and could suppress the proliferation and growth of osteosarcoma through the regulation of Eag1.Moreover,it may be a novel target for osteosarcoma therapy.

Objective To explore the expression of ether à go-go 1 ( Eag1) in human osteosarcoma and its molecular mechanisms of regulating the malignant phenotype of osteosarcoma .Methods The expression levels of Eag 1 in osteosarcoma cell lines and human osteosarcoma tissues were detected by reverse transcription polymerase chain reaction ( RT-PCR), western blot analysis and immunohistochemistry .The small interfering RNA ( siRNA) was used to inhibit the expression of Eag 1.The abilities of proliferation and invasion in osteosarcoma cells transfected with Eag 1 siRNA were determined by CCK-8, colony formation assay, transwell assay and wound healing assay .The osteosarcoma xenograft model of nude mouse was established and tumor growth curve was drawn .Western blot analysis was performed to detect the expression of vascular endothelial growth factor ( VEGF) and signal transducer and activator of transcription 3 ( STAT3) in osteosarcoma cells transfected with Eag 1 siRNAs.Results Eag1 was overexpressed in the osteosarcoma cells and tissues .Compared with the scrambled siRNA group , the cell survival rates of Eag 1 siRNA1 and Eag1 siRNA2 groups of the two cell lines were significantly lower .[ MG-63 cells: scrambled siRNA group (100.0±4.65)%, Eag1 siRNA1 group (63.57±3.89)%, and Eag1 siRNA2 group (54.13±3.70)%;Saos2-cells:scrambled siRNA group (100.00±5.46)%, Eag1 siRNA1 group (56.70±5.34)%, and Eag1 siRNA2 group (40.27±5.28)%(P<0.001 for all)].Similar results were obtained from colony formation assay .The colony formation rates of MG-63 cells: the scrambled siRNA group was (92.00±3.46)%, Eag1 siRNA1 group (60.00±3.06)%, and Eag1 siRNA2 group (53.67±2.40)%; the colony formation rates of Saos-2 cells: the scrambled siRNA group was (92.00±5.57)%, Eag1 siRNA1 group (52.33±5.13)%, and Eag1 siRNA2 group (41.67±27.3 )%.Compared with the scrambled siRNA group , P<0.001 for all.The tumor volumes of osteosarcoma xenograft in the Eag 1 siRNA1 and Eag1 siRNA2 groups were significantly smaller than that in the scrambled siRNA group after 10 days treatment (P<0.01 for all).The invasion assay data showed that MG-63 and Saos-2 cells transfected with Eag 1siRNAs exhibited the ability of cell invasion , when compared with the cells transfected with scrambled siRNA .( Invasive cell number of MG-63 cells:the scrambled siRNA group was 134.00 ±3.61, Eag1 siRNA1 group 105.20 ±2.52, and Eag1 siRNA2 group 91.00±3.01; Invasive cell number of Saos-2 cells: the scrambled siRNA group was 132.30±3.23, Eag1 siRNA1 group 114.30±3.48, and Eag1 siRNA2 group 82.67±6.33.Compared with the scrambled siRNA group, P<0.01 for all.The migration rates were (62.48±1.83)%, (35.98±1.23)%and (32.30±1.20)%in the three groups of MG-63 cells, and (70.15±1.42)%, (41.38±1.34)%and (32.40±1.92)%in the three groups of Saos-2 cells, respectively.Compared with the scrambled siRNA group , P<0.001 for all.Notably, the expression levels of VEGF decreased evidently after Eag 1 siRNAs transfection , paralleled with reduction in the expression levels of STAT 3.Conclusions Eag1 may promote osteosarcoma cell proliferation and invasion by targeting STAT 3-VEGF pathway and may be a potential therapeutic target for osteosarcoma .

Objective To explore the expression of ether à go-go 1 ( Eag1) in human osteosarcoma and its molecular mechanisms of regulating the malignant phenotype of osteosarcoma .Methods The expression levels of Eag 1 in osteosarcoma cell lines and human osteosarcoma tissues were detected by reverse transcription polymerase chain reaction ( RT-PCR), western blot analysis and immunohistochemistry .The small interfering RNA ( siRNA) was used to inhibit the expression of Eag 1.The abilities of proliferation and invasion in osteosarcoma cells transfected with Eag 1 siRNA were determined by CCK-8, colony formation assay, transwell assay and wound healing assay .The osteosarcoma xenograft model of nude mouse was established and tumor growth curve was drawn .Western blot analysis was performed to detect the expression of vascular endothelial growth factor ( VEGF) and signal transducer and activator of transcription 3 ( STAT3) in osteosarcoma cells transfected with Eag 1 siRNAs.Results Eag1 was overexpressed in the osteosarcoma cells and tissues .Compared with the scrambled siRNA group , the cell survival rates of Eag 1 siRNA1 and Eag1 siRNA2 groups of the two cell lines were significantly lower .[ MG-63 cells: scrambled siRNA group (100.0±4.65)%, Eag1 siRNA1 group (63.57±3.89)%, and Eag1 siRNA2 group (54.13±3.70)%;Saos2-cells:scrambled siRNA group (100.00±5.46)%, Eag1 siRNA1 group (56.70±5.34)%, and Eag1 siRNA2 group (40.27±5.28)%(P<0.001 for all)].Similar results were obtained from colony formation assay .The colony formation rates of MG-63 cells: the scrambled siRNA group was (92.00±3.46)%, Eag1 siRNA1 group (60.00±3.06)%, and Eag1 siRNA2 group (53.67±2.40)%; the colony formation rates of Saos-2 cells: the scrambled siRNA group was (92.00±5.57)%, Eag1 siRNA1 group (52.33±5.13)%, and Eag1 siRNA2 group (41.67±27.3 )%.Compared with the scrambled siRNA group , P<0.001 for all.The tumor volumes of osteosarcoma xenograft in the Eag 1 siRNA1 and Eag1 siRNA2 groups were significantly smaller than that in the scrambled siRNA group after 10 days treatment (P<0.01 for all).The invasion assay data showed that MG-63 and Saos-2 cells transfected with Eag 1siRNAs exhibited the ability of cell invasion , when compared with the cells transfected with scrambled siRNA .( Invasive cell number of MG-63 cells:the scrambled siRNA group was 134.00 ±3.61, Eag1 siRNA1 group 105.20 ±2.52, and Eag1 siRNA2 group 91.00±3.01; Invasive cell number of Saos-2 cells: the scrambled siRNA group was 132.30±3.23, Eag1 siRNA1 group 114.30±3.48, and Eag1 siRNA2 group 82.67±6.33.Compared with the scrambled siRNA group, P<0.01 for all.The migration rates were (62.48±1.83)%, (35.98±1.23)%and (32.30±1.20)%in the three groups of MG-63 cells, and (70.15±1.42)%, (41.38±1.34)%and (32.40±1.92)%in the three groups of Saos-2 cells, respectively.Compared with the scrambled siRNA group , P<0.001 for all.Notably, the expression levels of VEGF decreased evidently after Eag 1 siRNAs transfection , paralleled with reduction in the expression levels of STAT 3.Conclusions Eag1 may promote osteosarcoma cell proliferation and invasion by targeting STAT 3-VEGF pathway and may be a potential therapeutic target for osteosarcoma .

Objective:To investigate the antibacterial effect of iodophor on Staphylococcus aureus biofilm (BBF).Methods:Staphylococcus aureus were cultured in vitro and 480 pieces of titanium alloy plates were selected. On the surface of titanium plates, in vitro models of Staphylococcus aureus biofilms were established at days 7, 14, 21 and 28 respectively with 120 pieces of titanium plates at each time points. The biofilms at each time point were assigned to no iodophor immersion (PBS group), 5 g/L iodophor immersion for 5 minutes (5-min group) and 5 g/L iodophor immersion for 10 minutes (10-min group), according to the random number table method. FITC-ConA, propidium iodide (PI) and SYT09 were used to dye Staphylococcus aureus in PBS group. After dyeing, confocal laser scanning microscopy and scanning electron microscopy were used to observe the morphological structure of bacterial biofilms, and the Colony forming unit (CFU) was counted by the viable count method. In the other two groups, PI and SYT09 were applied to dye Staphylococcus aureus, and then confocal laser scanning microscopy and scanning electron microscopy were used to observe the changes of biofilms and bacterial viability after iodophor immersion. The antibacterial effect of iodophor was evaluated by the viable count method.Results:After dyeing Staphylococcus aureus with FITC-ConA and PI in PBS group, confocal laser scanning microscopy showed that the extracellular polymers of the bacteria increased gradually with the extension of culture time. The space structure of biofilm was gradually mature, changed significantly at day 21 and became mature at day 28. After staining Staphylococcus aureus with PI and SYT09 in PBS group, confocal laser scanning microscopy showed that the number of bacteria increased, and had a mountain-like shape. Scanning electron microscopy showed that the number of bacterial extracellular polymers increased gradually with the extension of culture time and a structured microenvironment was formed and gradually matured. In 5-min and 10-min groups, all bacteria were killed at days 7 and 14 [0(0, 0)CFU/ml], the antibacterial effect was weakened at 21 days, but the antibacterial effect of iodophor immersion in 10-min group [100 (100, 125)CFU/ml] was better than that in 5-min group [300 (275, 425)CFU/ml] ( P<0.05). There was no significant difference in iodophor immersion in 5-min group [500 (375, 700)CFU/ml] and 10-min group [250 (175, 400)CFU/ml] at 28 days ( P>0.05). Conclusions:The maturation of biofilm is the overall maturation of bacteria and bacterial extracellular polymers and the formation of a spatialized microenvironment. Bounded by the 21st day, biofilms are divided into young biofilms and mature biofilms. The main difference between them lies in the maturation of extracellular polymers and microenvironment. For the bacterial biofilm with culture time less than 21 days, the antibacterial effect of the iodophor immersion for 10 min is better than that of 5 min. However, for the bacterial biofilm with culture time greater than 21 days, there is no significant difference in the antibacterial effect of the bacterial biofilm of prolonged iodophor immersion time.

We report the diagnosis and treatment of a rare case of epidural analgesia failure followed by postpartum subdural hematoma. The patient underwent vaginal delivery under epidural analgesia at 32 +6 gestational weeks due to threatened premature labor, during which an unexpected dural rupture occurred. She gave no history of headache and there was no obvious abnormality during the pregnancy. However, on postpartum day 4, the patient complained of headache that could not be relieved when supine, but without any other neurological symptoms. A prompt cranial CT examination showed a left frontotemporal subdural hematoma. After conservative management with intravenous drip of mannitol, re-examination of cranial CT showed that the left frontotemporal subdural hematoma was mostly absorbed and the patient was discharged on postpartum day 18. The patient was healthy during follow up. Intracranial subdural hematoma after dural puncture is a rare and serious complication that requires early recognition and treatment.

We report the induced labor of conjoined twins in the second trimester in a woman with a history of two previous cesarean sections, the last one of which was performed in 2017. This 25-year-old patient was found to have thoracolumbar conjoined fetuses with one heart and polyhydramnios through the routine ultrasound examination at 19 +5 gestational weeks and was admitted at 20 +1 gestational weeks. After a full assessment of the fetal and maternal condition through multidisciplinary consultation, it was determined to attempt a vaginal delivery as no absolute contraindication for induction of labor. The patient was given 300 mg mifepristone orally plus an amniotic cavity injection of 100 mg ethacridine lactate. Regular contractions occurred 28 hours after medication. The patient delivered a pair of dead female conjoined twins at 20 +6 gestational weeks following successful induction of labor, with an assisted vaginal breech delivery. There was no soft tissue damage in the birth canal, and the estimated blood loss was 150 ml. Pathological examination and autopsy showed thoracolumbar conjoined deformity twins with a common heart and liver. Adequate prenatal evaluation, a detailed understanding of the indications for induction of labor and vaginal delivery, closed monitoring during labor, and preparation for emergency cesarean section, are essential safety measures for induced labor of conjoined twins in women with a scarred uterus in the second trimester.