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 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 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.

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.

Objective To investigate the possibility of interventional treatment of placenta accreta. Methods One patient with placenta accreta was treated with uterine arterial infusion of cef and MTX followed by embolization. Results Her placenta discharged through vagina at the 27th day after uterine aterial embolizayiea. No remnant was found in the uterine by Doppler imaging. The patient uneventfully recovered.Conclusions Intervetional therapy with uterine arterial embolization is a new and reliable method for treatment of placenta accheta.

Neuropathic pain(NP)is directly caused by lesions or diseases of the somatosensory system.It can be attributed to a variety of causes and has a complex pathological mechanism.Purinergic P2X receptors play a critical role in NP.P2X receptors will be activated by the extracellular adenosine triphosphate(ATP)to mediate the pain when the tissue is damaged.In vertebrates,there are 7 subtypes of P2X receptors(P2X1-7)encoded by genes,among which the receptors of P2X3,P2X4,and P2X7 are closely related to NP.Acupuncture produces good therapeutic efficacy on NP.To explore the mechanism of purinergic signaling involved in acupuncture analgesia,a review of the actions of P2X3,P2X4,and P2X7 receptors in acupuncture for NP was conducted.The P2X3 receptor is specifically expressed in small-and moderate-sized neurons in the dorsal root ganglion of the primary afferent nerve and trigeminal ganglion.Acupuncture can directly mediate analgesia by down-regulating the expression of P2X3 receptor in neurons.P2X4 and P2X7 receptors mainly exist in the spinal microglia.Acupuncture can inhibit the nociceptive transmission of substantia gelatinosa neurons by regulating P2X4 receptor in spinal microglia.Moreover,acupuncture may alleviate NP by inhibiting the secretion of inflammatory factors by reducing P2X7 receptor in the spinal microglia.

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.

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.