Objective To study the effect of silencing Nek2 via RNAi on cell cycle of ovarian cancer SKOV 3 cells and the re‐lated molecular mechanism .Methods The Nek2‐siRNA was transfected into the ovarian cancer SKOV3 cells .The change of cell cy‐cle of SKOV3 cells at 48 h after transfection was examined by the flow cytometry technique ;Western blot assay was used to deter‐mine the change of level of the cell cycle related factors cyclinB1 ,CDK1 ,P27 and the phosphorylation level of the ERK1/2 after Nek2‐siRNA transfection for 48 h .Results The flow cytometry detection results showed that the proportion of the cells in G 2/M stage in the blank control group ,negative control group and RNAi group was 13 .72% ,12 .27% and 1 .56% respectively .Compared with the control group ,the number of the cells in G2/M stage in the transfected group was reduced obviously (P< 0 .05) .The Western blot detection results showed that compared with the control group ,the expression of cyclinB1 and CDK1 protein in SK‐OV3 cells was significantly reduced ,the expression of P27 was increased after silencing Nek2 and the phosphorylation level of ERK1/2 in SKOV3 cells was significantly reduced after silencing Nek2 gene(P< 0 .05) .Conclusion Silencing Nek2 gene might block the ovarian cancer cell line SKOV 3 initiating mitosis ,thus inhibit their proliferation .

Objective To investigate the effect of recombinant human erythropoietin on immune function in mice with multiple myeloma.Methods Multiple myeloma model mice were divided randomly into control group and treatment groups (low,middle and high doses).The rats in treatment groups were injected with recombinant human erythropoietin 100 μL (2.5,5.0 and 10 mg/kg)via the tail vein for 30 days,and the rats in control group were injected isotonic saline solution of the same volume instead.After the experiment,the colorimetric MTT assay,neutral red method and ELISA kit were used to evaluate the proliferation of B and T lymphocytes,the phagocytic ability of macrophages and the content of serum TNF-α.Results Compared with those in control group,different doses of recombinant human erythropoietin could significantly increase the proliferation of lymphocytes,increase the phagocytosis of macrophages and improve the production of serum TNF-α(P < 0.05 ). Conclusion Recombinant human erythropoietin can significantly improve immunoregulation of mice with multiple myeloma.

Objective To explore the neutral grain cell gelatinase associated lipid lipocalin(NGAL ) and β2 microglobulin (β2‐MG) predictive value of diabetic nephropathy(DN) .Methods From January 2015 to April 2016 in the hospital for 78 cases of DN patients recorded as the observation group ,and then 50 cases of healthy volunteers were choosen as the control group at the same period .NGAL ,β2‐MG ,BUN ,SCr and GFR levels were compared in the two groups .And the positive rate and correlation were ana‐lyzed .Results The levels of NGAL and β2‐MG in the observation group were(473 .21 ± 127 .09)ng/mL ,(2 .76 ± 0 .45)mg/L , which were significantly higher than that in the control group(61 .36 ± 14 .230)ng/mL ,(0 .81 ± 0 .14)mg / ,respectively .The posi‐tive rate of NGAL and β2‐MG in the observation group were 96 .15% (75/78) ,91 .03% (71/78) ,which were significantly higher than that of the control group of 2% (1/50) ,6% (3/50) .While the BUN ,SCr in observation groups were significantly higher than that in the control group ,while the level of eGFR was significantly lower than that of the control group ,the differences were statis‐tically significant(both P< 0 .05) .NGAL ,β2‐MG and BUN ,SCr showed positive correlation(R = 0 .812 ,P= 0 .000 ;R = 0 .728 ,P=0 .001 ;R = 0 .748 ,P= 0 .001 ;R = 0 .685 ,P = 0 .021 ) ,and eGFR showed a negative correlation (R = - 0 .415 ,P = 0 .000 ;R=- 0 .371 ,P= 0 .000) .Conclusion The level of NGAL and β2‐MG in DN patients is significantly increased ,NGAL ,β2‐MG levels are significantly correlated with BUN .

Objective To investigate the effect of ulinastatin on renal function and oxygen metabolism in brain injury patients . Methods 50patientswithcraniocerebralinjuryinthehospitalwereenrolledinthestudy,andwererandomlydividedintocontrol group(received conventional craniocerebral injury treatment ,25 cases) and observation group(received conventional craniocerebral injury treatment and ulinastatin treatment ,25 cases) .Then the renal function and oxygen metabolism related indicators of the two groups before the treatment and at third ,seventh and fourteenth day after the treatment were compared .Results The renal function and oxygen metabolism related indicators of observation group at third ,seventh and fourteenth day after the treatment were all ob‐viously better than those of control group ,the differences of the two groups after the treatment were significant(P<0 .05) .Conclu‐sion The effect of ulinastatin on the renal function and oxygen metabolism of patients with craniocerebral injury are great ,and it has positive clinical significance for the improvement of patients′body function state .

Objective To test whether feeder cells derived from mouse embryonic stem cells(mESCs) could support the growth of mESCs themselves. Methods mESCs were induced to form mouse embryoid bodies(EB),and then fibroblast-like cells were derived from further differentiated mEB(mEB-dF),which served as feeder cells.The undifferentiation of mESCs grown on mEB-dF was confirmed by morphological analysis,colony efficiency and cell differentiation rate of mESCs,immunocytochemistry,alkaline phosphatase staining and RT-PCR.The pluripotency of mESCs grown on mEB-dF was examined by RT-PCR,inducing their differentiation in vivo and in vitro. Results(Forty-eight) fibroblast-like cells lines were derived from the same EB at three periods(d 10,d 15 and d 20),and five of them,mostly derived from d15 EB,were able to maintain mESCs in undifferentiated status and pluripotential ability over 10 passages.mESCs cultured on these feeder cell lines expressed alkaline phosphatase and specific mESCs markers,including SSEA-1,OCT-4,NANOG,and formed EB in vitro and teratomas in vivo.However,the majority of mEB-dF lines(43/48) has no such ability. Conclusion This study not only provides a novel feeder system for mESCs culture,avoiding lot of disadvantages of mouse embryo fibroblasts used as the feeder,but also indicates that fibroblast-like cells derived from mESCs take on different functions.The molecular mechanism of different function of these fibroblast cells is worthy of further investigations.

Problem-based and student-centered learning patten brings new challenges to the both sides of teaching and learning.Clinical probation is the first step of medical students.The problem-based learning patten is used to clinical probation and achieves good results.It helps to improve medical students' quality and should be applied in China medical education.

Objective To explore the impact of maximal movement on the stabilization of the four corner arthrodesis.Methods Flexion,extension,ulnar deviation and radial deviation movements were simulated on the wrists of 21 fresh cadaver fore-arm specimens.Maximal movements were performed in each of the four directions. Firstly,anteroposterior view and lateral view photos were taken before and after movement in each direction.Sec- ondly,capito-lunate angle,radio-lunate angle,captio-lunate posteroantefior angle,the height and width of the syzygium were measured before and after movement in each direction.The wrist before each kind of movement was used as the control for itself after the movement in the test.Results Compared with the control data,the differ- ences in capito-lunate angle,radio-lunate angle and the height of the syzygium were statistical significant before and after movement in the direction of flexion 40?and extension 30?(P＜0.05).Moreover,the differences in the captio-lunate posteroanterior angle and the width of the syzygium were also statistical significant before and after movement in radial deviation 15?and ulnar deviation 20?(P＜0.01).Maximal movements at other positions had little impact on the stability of the arthrodesis.Conclusion It is safe to perform high intensity movements to the four-comer syzygium when the range of motion is not beyond flexion 40?,extension 30?,ulnar deviation 20?and radial deviation 15?.Cireumduction should be avoided.

0.05). In bone defects PeroGlas granales bond to bone and new bone formation were observed 4 weeks after operation. The bone defects were repa ired by new bone 12 weeks after PerioGlas implantation. Conclusion: PerioGlas may induce new bone formation in osseous defects.

Objective To study the effects of knock down midkine(MK)by siRNA on chemosinsitivity in bladder cancer cells. Methods Three MK siRNAs were designed and constructed. After transfected with MK siRNA or scrambled siRNA for different time, cultured cells were harvested to carry on the next experiments. Expression of MK was determined by real-time RT-PCR and western blot, and apoptosis were evaluated by caspase-3 activity and TUNEL assay. MTT was performed to examine the inhibition effect of Paclitaxel (PTX) on cells. Results MK siRNA could down-regulate the MK expression in a dose- and time-dependent manner. The MTT results showed that the inhibit rates were (18.21±0.36)%, (18.19±0.29)%, (17.89±0.33)%, (1.86±0.52)%, (32.56±0.53) %, (53. 83±0.38) % and (78. 95±0.55) % in different groups PTX alone(0.2μmol/L), ConA +PTX(0.2 μmol/L), Con-B +PTX(0.2 μmol/L), siRNA alone(12. 50 nmol/L), siRNA(3. 125nmol/L) + PTX (0. 2 μmol/L), siRNA (6. 25 nmol/L) + PTX (0. 2 μmol/L) and siRNA (12. 50nmol/L)+PTX(0.2 μmol/L), respectively. The TUNEL results showed that apoptosis index was (1.81 ±0. 36)%, (1. 89±0. 38)%, (5. 56±0. 58)%, (9. 68±0.55)% and (15. 25±0.56)% in different groups (Con-A, Con-B, siRNA (3. 125 nmol/L), siRNA (6. 25 nmol/L) and siRNA ( 12. 5nmol/L), respectively. The activity of caspase-3 increased significantly in transefected cells with a dose-and time-dependent manner. Conclusion MK siRNA could sensitize human bladder cancer cells to chemotherapy which might be through the apotosis.

Objective To evaluate the clinical results of self-designed anterior cervical low-profile plate system (ACLPS). Methods The anterior cervical plate system, composed of one plate, two locking caps and two types of screws. The system was made of medical titanium alloy possessing the following characteristics: low profile, instant locking, versatile use and unicortical screw fixation. 37 patients with cervical disorders were fixed with ACLPS after anterior spinal decompression and interbody fusion. The diagnosis included spondylosis in 8 cases, cervical intervertebral disk protrusion in 12, fracture and /or dislocation with paralysis in 14 and metastatic tumor in 3. The fusion were performed with autologous iliac crest bone graft for 29 patients while the others with cages. The fixed levels ranged from C3 to T1. Results 37 cases were followed-up for an average of 9.6 months, ranging from 6 to 12 months. Postoperatively all patients felt comfortable in neck without any foreign body sensation. No obvious complications such as wound infection, hematoma, asphyxia, nerve injury, laceration of spinal cord dura, leakage of cerebrospinal fluid, bleeding and nonhealing of the wound were observed. Solid interbody fusion were achieved in all cases. According to JOA scores, the total effective rate was 94.6% while the good rate was 83.8%. The radiograph confirmed that there was no dislocation or subsidence of graft, no obvious loss of intervertebral height, no loosening or failure of implant. Conclusion In regard to the high rigidity, good bony purchasing and easy handling, ACLPS is able to provide sufficient biomechanical stability and is an optional choice for clinical use.