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.

Animals ; Humans ; Hypertension, Pulmonary ; complications ; Hypothermia ; complications ; Hypoxia ; etiology ; therapy ; Respiratory Distress Syndrome, Adult ; complications ; Shock, Septic ; complications

Objective　To study the response rate and toxic side effect of Vindesine, aclarubicin and mitomycin (VAM) as a new regime of second line chemotherapy for ovarian carcinoma refractory to platinum group of drugs.Methods　From June 1997 through August 1998, 25 cases of refractory ovarian carcinoma were treated with VAM regime. The response rate and the side reactions were analyzed. Results　The overall response rate was 32.0%, with 3 complete response (CR) and 5 partial response (PR). In the platinum refractory, platinum-resistant platinum-senstitive strata, the response rates were 18.2% 28.6% and 57.1% respectively. Myelosuppression was the main toxic effect. Grade 3 and grade 4 neutropenia occurred in 40.0% and thrombocytopenia in 28.0% of the patients. Alopecia occurred in 32% of cases. Gastrointastinal, cardiac and neurologic toxicities were mild. Conclusion　VAM regime is a practical and effective second-line chemotherapy for patients with ovarian carcinoma refractory to platinum based chemothrapy.

Objective: To explore the nature of voltage dependent ion channels and basic electrophysiological characteristics of cochlear spiral ganglion neurons of apical and basal turn by patch clamp techniques of whole cell configure on murine spiral ganglion neurons. Method: Different voltage dependent ionic currents were recorded with patch clamp techniques of whole cell configure on the condition of different internal electrode solution,blockers and stimulus protocol. Result: Inward sodium channel current(I_(Na)), hyperpolarization-activated inward cationic current(I_h) ,outward delay rectification potassium current (I_k) and outward transient potassium current (I_A)were recorded , significant difference of electrophysiological characteristics of I_A and I_k was found between apical and basal turns(P＜0.05). Conclusion: Various ionic currents are recorded,which shows that spiral ganglion neurons have the base of ionic channels to complete formation,conduction and modulation of action potential for auditory information transduction, the difference of electrophysiological characteristics between apical and basal turns contributes to the course of hearing formation.

A simple and sensitive fluorimetric method is proposed for the determination of Cr(Ⅵ).The method is based on the oxidation of Cr(Ⅵ) to the pyronine in the sulfuric acid medium.The detection limit for Cr(Ⅵ) is 2.2μg/L,the linear range of the determination is 8.0～88μg/L.The method has been used to determine Cr(Ⅵ) in electroplating waste,electroplating solution and alloy steel samples with satisfactory results.

Objective To evaluate the short term results of shoulder hemi-arthroplasty. Methods Twenty-one patients with comminuted intra-articular fracture of proximal humerus underwent Neer type (Link, Germany) humeral head replacement from August 2000 to January 2004, which involved 5 males and 16 females,with an average age of 70 years (range, 64-76 years). According to AO M?ller classification, 3 cases were of C1 type, 8 C2 type, and 10 C3 type. The important steps of the procedure included insertion of the prosthesis in 40 degree retroversion and reconstruction of the rotary cuff of shoulder. The rehabilitation training course was divided into three stages. The patients were trained passively in the first stage (1 day-6 weeks); The muscles around the shoulder undertook isometric contraction once the bone fusion of greater and lesser tuberosity were obtained on X-ray for the second stage (7 -12 weeks); The improvement of the muscle strength and ROM of the shoulder were targeted throughout the third stage (13-24 weeks). All cases were followed up with Neer's scoring system comprising the symptoms, function, range of motion (ROM) of shoulder, and position of the prosthesis. Results The follow-up was performed twice a month within one year postoperatively, and one time three months after that.The mean duration of follow-up was 34.5 months (range, 14 to 55 months). Only slight pain was complained occasionally in 4 cases. No difficulty in active daily life was found in 21 cases .The average ROM of shoulder was flexion 115?(range, 65?-165?),extension 42.5?(15?-70?),abduction 85?(50?-120?), adduction 32.5?(20?-45?), external rotation 40?(20?-60?),and internal rotation 50?(10?-90?). No dislocation or subluxation of shoulder occurred in all cases. Assessed by Neer's scoring system, 8 cases were rated as excellent, 9 as good, 3 as fair and 1 as poor. The excellent and good rate was 80.9%. The patients' satisfactory rate was 90%. Conclusion The shoulder hemi-arthroplasty show good short term results for comminuted intra-articular fractures of the proximal humerus. The key points to improve the postoperative results are presented as follows: less invasive technique, appropriate retroversion angle for humeral head prosthesis, very solid reconstruction for rotary cuff of shoulder, and correct consecutive physical therapy after surgery.