Objective To investigate the impact of LY294002 on the proliferation of cancer stem cell-enriched spheroid cells from human hepatocellular carcinoma via regulating phosphatidylinositol-3-kinaseprotein kinase B(PI3K-Akt)signaling pathway. Methods The cancer stem cell-enriched spheroid cells were genera-ted by culturing HepG2 cells in serum-free medium. LY294002(10,20,30 μmol/ L),an inhibitor of PI3K-Akt signaling pathway,was used in the experimental groups,without used in the control group. The impact of LY294002 on the spheroid cells proliferation was confirmed by cell counting kit(CCK-8 kit). The expression of Akt was tested by Western blotting. The expression of PI3K-Akt signaling pathway downstream genes such as decoy receptor 3(DcR3),mammalian target of rapamycin(mTOR),B-cell lymphoma(Bcl)-2 and Cyclin D1 were tested by real-time PCR. Results 30 μmol/ L LY294002 could inhibit the proliferation of spheroid cells, and significant difference in the absorbance(A value)was observed between the experimental group and control group[(0. 14 ± 0. 03)vs(0. 56 ± 0. 01),t = - 8. 915,P = 0. 000]. The expression level of phosphorylated Akt protein increased[(0. 57 ± 0. 08)vs(0. 16 ± 0. 42),t = 6. 027,P = 0. 026]. The mRNA of DcR3 [(0. 38 ± 0. 08)vs 1,t = 13. 060,P = 0. 006],mTOR[(0. 37 ± 0. 04)vs 1,t = 30. 363,P = 0. 001],Bcl-2 [(0. 26 ± 0. 04)vs 1,t = 33. 554,P = 0. 001]and Cyclin D1[(0. 10 ± 0. 02)vs 1,t = 63. 528,P = 0. 000] decreased. Conclusion LY294002 could inhibit the proliferation of cancer stem cell-enriched spheroid cells from human hepatocellular carcinoma via inhibiting PI3K-Akt signaling pathway.

Objective To investigate the changes in the number of synapses and neurons in the spinal dorsal horn in a rat model of phantom limb pain. Methods Eleven healthy adult SD rats of both sexes weighing 209-300 g were randomly divided into 2 groups: sham operation group (group S, n = 5) and phantom limb pain group (group P, n = 6). Phantom limb pain was induced by resection of a 0.5 cm segment of unilateral sciatic nerve in group P. In group S unilateral sciatic nerve was exposed but not transected. The animals were observed for autotomy and scored (0 = no autotomy, 13 = the worst autotomy) after operation and were sacrificed on the 28th day after operation. The L3-6 segment of the spinal cord was removed for determination of the number of neurons (by Nissl's staining) and synapses (by synaptophysin immuno-histochemistry).Results In group S no animal developed autotomy. In group P autotomy started from the 2nd day after operation and the score reached 9-11. The number of the neurons in the spinal dorsal horn in all 4 segments and the number of synapses in L3 and 16 segments were comparable between the two sides and the 2 groups. The number of synapses in the spinal dorsal horn of L4and L5 segment was significantly larger in the operated side than in the contralateral side in group P. Conclusion The number of synapses in the spinal dorsal horn significantly increases in animals with plantom limb pain which induces no increase in the number of neurons in the spinal dorsal horn.

Objective To study the effect of the combined medial and lateral operative approach for the serious intra -articular fractures of calcaneus.Methods The lateral and medial approach for the treatment of the serious intra -articular fractures of calcaneus 22 cases (26 feet),fully exposed in the medial and lateral subtalar joint surface,reduct subtalar joint surface,drill into Kirschner nails for reduction of the calcaneus angle,then graft bones fully in the defect left after reduction,used calcaneus anatomic plate and screws to fix the fracture on the lateral. Results The patients were followed up for 10 -22 months.According to the American Orthopaedic Foot and Ankle association ankle scoring system,the averange score was 87 points.Normal union of 22 feet,4 feet delayed union,skin necrosis no incision,no infections,no cases of neurovascular injury.Conclusion The combined medial and lateral operative approach for the serious intra -articular fractures of calcaneus is an effective method,and can fully expose the lateral and medial fracture,and is conducive to reduction of the lateral and medial articular surface,and restoration of calcaneus shape,reduce malunion and the foot -pain with walking.

Objective To investigate the changes in the number of microglias and astrocytes in the spinal dorsal born in a rat model of phantom limb pain.Methods Eleven healthy adult SD rats of both sexes weighing 290-300 g were randomly divided into 2 groups:sham operation group (group S,n =5 ) and unilateral sciatic nerve transection group (group SNT,n =6).Phantom limb pain model was induced by resection of a 0.5 cm segment of unilateral sciatic nerve in group SNT.In group S unilateral sciatic nerve was exposed but not transected.The animals were observed for autotomy and scored (0 =no autotomy,13 =the worst autotomy) after operation and were sacrificed on the 28th day after operation.The L5 segment of the spinal cord was removed for determination of the number of microglials (by iba-1 immuno-histochemistry) and astrocytes (by GFAP immuno-histochemistry).Results In group S no animal developed autotomy.In group SNT autotomy started from the 2nd day after operation and the score reached 9-11.Compared with group S,the number of the microglias and astrocytes in the spinal dorsal horn was significantly decreased in the operated side in group SNT ( P ＜ 0.05 ).Conclusion The number of microglias and astrocytes in the spinal dorsal horn is decreased in animals with phantom limb pain.

Objective To investigate the effect of different bone cement volume injected after vertebroplasty on stress distribution of the upper and lower endplates of the injured vertebra in the flexion and extension positions by finite element analysis.Methods Two patients with osteoporotic compression fractures of L2 vertebral body were chosen.Bone cement volumes of 25％ and 50％ were respectively injected into the fractured vertebral body bilaterally.Pre-and post-operative finite element models of fractured and adjacent vertebral bodies were developed after extracting the CT data.Stress changes at L2 superior-and inferior-vertebral endplates were observed in the vertical,flexion,and extension loading.Results Stress at the upper and lower endplates of the fractured vertebra increased after bone cement injection and more increase was observed in 50％ bone cement group than in 25％ bone cement group.In the neutral position,the stress mainly concentrated at posterior vertebral body,with stress of the upper endplate improving from 0.91 × 106 N · mm to 0.93 × 106 N · mm and stress of the lower endplate from 0.98 × 106 N ·mm to 1.03 × 106 N · mm.In the anterior flexion,the stress mainly concentrated at anterior vertebral body,with stress of the upper endplate improving from 4.44 × 106 N · mm to 5.55 × 106 N · mm.In the posterior extension,the stress mainly concentrated at medial vertebral body,with stress of the upper endplate improving from 21.95 × 106 N · mm to 22.10 × 106 N · mm and stress of the lower endplate from 30.60 ×106 N · mm to 30.88 × 106 N · mm.The finite element model revealed stress at the upper and lower endplates was higher in the anterior flexion and posterior extension than in the neutral position,while in the anterior flexion the stress was comparatively higher.Conclusions A small amount of implanted bone cement act no evident effect on stress distribution of the upper and lower vertebral endplates.However,the stress changes are significant as cement volume reached up to 50％,especially in the flexion position,as may suggest an enhanced risk of adjacent intervertebral disc degeneration and adjacent vertebral body fractures.

To determine the effect of straight-leg-raising (SLR) movement on epidural fibrosis after laminectomy, 40 adult New Zealand rabbits were selected as laminectomy models in the study. They were divided into 2 groups: a SLR group (group S) and a control group (group C) randomly, with each group having 20 animals. All rabbits were subjected to total laminectomy in the site of S1. Every 5 rabbits in each group selected randomly were killed at the 1st, 2nd, 4th, and 8th week after the surgery. Segments of spines from L7 to S2 were removed en bloc. After gross evaluation, specimens were sliced up. The slices were stained by HE and Masson's trichrome methods respectively for histological examination. Our results showed that formation process of scar in group S was retarded as compared with that of group C at the time of the 2nd-week, but there was no statistical difference between groups in the adhesion degree (P> or =0.05). At the 4th and 8th week, the epidural fibrosis of group S was more serious than that of group C. Since the 2nd-week, the area of scar in group S was larger than that of group C. The number of fibroblasts and inflammatory cells in group S were larger than those of group C at early stage. But in later stage, there was no statistical significance between the two groups. It is concluded that SLR movement after laminectomy may promote the formation of epidural fibrosis and retard the maturity of scar. SLR movement can also aggravate scar adhesion.

Objective To investigate the effect of serum CYFRA21-1 and SCCAg levels on the prognosis of patients with cervical cancer radiotherapy.Methods 100 cases of cervical cancer were diagnosed by pathological examination in our hospital from October 2008 and October 2011,Using radioimmunoassay(RIA) in detection of serum CYFRA21-1 and SCCAg level,Pearson and Spearman correlation is used to study the relationship between serum CYFRA21-1 and SCCAg level and cervical cancer patients with clinical features,using the Cox regression model of CYFRA21-1 and SCCAg level and uterine cervix cancer death or recurrence and metastasis of the relationship.Results 100 cases of patients with cervical cancer,endpoint events in 21 cases,LNR4 patients appear higher risk of endpoint events about 44.3%,CYFRA21-1 and SCCAg levels were with cervical cancer patients with CA-125 values,tumor staging,depth of tumor invasion into positive correlation,and the age of patients,no significant correlation,tumor differentiation(P=0.013),serum CYFRA21-1 levels(P=0.02),serum SCCAg levels(P=0.03),depth of tumor invasion(P=0.04) is cervical cancer patients with endpoint events independent risk factors.Conclusion CYFRA21-1 and SCCAg levels were closely related to the prognosis of patients with cervical cancer radiotherapy.

Objective To discusses the clinical application value and safety of endoscopic minimally invasive cholecystolithotomy in gallstones treatment .Methods To retrospectively analyzes the clinical data of 94 patients with gallbladder stone from Feb .2010 to Feb .2013 ,and divided into endoscopic minimally invasive cholecystolithotomy (EMIC) group (46 cases) and laparoscopic chole-cystectomy (LC) group (48 cases) .Observed two groups of operation time ,intraoperative blood loss ,intraoperative bile duct inju-ry ,anal exhaust time ,postoperative hospitalization time ,bile reflux gastritis ,abdominal distention ,diarrhea ,common bile duct calculi and hospitalization expenses ,etc .Results Two groups of operation time ,hospitalization expenses ,length of hospital stay ,and intra-operative blood loss have no obvious difference(P>0 .05) ,in EMIC set ,the exhaust time ,postoperative bile duct calculi incidence , intraoperative bile duct injury ,bile reflux gastritis incidence and the incidence of abdominal distension ,diarrhea are significantly less than LC group (P<0 .05) .Conclusion The endoscopic minimally invasive cholecystolithotomy compared with laparoscopic chole-cystectomy had high security ,light pain ,quicker recovery ,less complications advantages .It has already achieved the purpose of min-imally invasive treatment ,while maintaining the integrity of bile duct and gallbladder function ,thus it is worthy of promoting .

Objective To discusses the feasibility of selective mini-cholecystolithotomy in treatment of gallstones .Methods To retrospectively analyzes the clinical data of the function good gallstones patients who treatmented by mini-cholecystolithotomy .Re-sults 4 cases treatmented by Laparoscopic cholecystectomy because of the severe cholecystitis ,1 case treatmented by laparoscopic cholecystectomy because of the severe adhesion around the gallbladder .51 cases treatmented by mini-cholecystolithotomy .In the fol-low-up of 1-3 years ,1 case of recurrence ,and the recurrence rate was 1 .96% .Conclusion The mini-cholecystolithotomy is a safe , effective ,feasible ,minimally invasive treatment method in treatmented gallstones ,but should be strictly grasp the surgical indica-tions .

Objective To determine the diagnostic value of double-phase enhancement and virtual endoscopy with multi-slice spiral CT (MSCT) on clinical staging of preoperative bladder cancer.Methods Seventy-five patients with bladder cancer diagnosed by fibercystoscope or operation. All of them were examined by double-phase enhancement and virtual endoscopy with MSCT. The images were analyzed and clinical staging were obtained. The findings of MSCT (71 cases)were compared with the post-operative histopathological results. Results There were 94 lesions to be found. The staging of MSCT: T1 26 cases,T2, 27 cases, T2b 13 cases, T3 12 cases, T4 16 cases. Histopathological results: pT1 28 cases, pT2a 24 cases, pT2b 14 cases, pT3 12 cases, pT4 16 cases. The sensitivity of preoperative staging on bladder cancer was 89.4 % (84/94) by double-phase enhancement of MSCT;the sensitivity of virtual endoscopy was 96.6% (84/87)for polyploidy tumors and 90. 9 % (10/11) for sessile lesions. When double-phase enhancement and virtual images were evaluated together, the sensitivity rate increased to 94.5%. When the tumors were confined within the bladder wall (≤T2b), the diagnostic accuracy of double-phase enhancement and virtual images was 91.2% (51/56). When the tumors had invaded the tissues and organs beyond the bladder wall (≥T3), the accuracy was 100% (28/28). Conclusion Double-phase enhancement and virtual endoscopy of MSCT is of great value in clinical staging of bladder cancer.