OBJECTIVETo investigate whether bortezomib can enhance the sensitivity of human prostate cancer (PCa) cells to natural killer (NK) cell-mediated cytotoxicity, and whether it produces the same effect on different PCa cell lines.

METHODSWe treated androgen-dependent PCa LNCaP cells and androgen-independent PCa DU145 cells with bortezomib at the concentrations of 0, 5, 10, 15, 20 and 25 nmol/L for 24, 48 and 72 hours, and then detected the proliferation and apoptosis of the tumor cells by CCK-8 and Annexin V/PI, respectively.

RESULTSThe proliferation rates of the DU145 cells treated with 15, 20 and 25 nmol/L bortezomib were (82.79 +/-2.04)%, (73.59+/- 2.95)% and (74.16+/- 6. 16)% at 48 hours and (71.24+/- 5.30)%, (51.20+/- 2.91)% and (38.02+/- 2.67)% at 72 hours, and those of the LNCaP cells were (77.04+/- 7.74)% , (42.61 +/- 6.62)% and (23.85 +/-6.04)% at 48 hours and (36.45 +/-7.02)%, (14.94 +/-5.76)% and (11.65 +/-5. 87)% at 72 hours, both significantly inhibited as compared with the control group (P <0.05). At 24 hours, the apoptosis rates of the DU145 cells treated with 15, 20 and 25 nmol/L bortezomib were (14.41 +/- 1.32)% , (16.13 +/- 1.55)% and (14.48 +/- 1.42)% , and those of the LNCaP cells treated with 20 and 25 nmol/L bortezomib were (12.77 +/- 1.28)% and (14. 84 +/- 1.65)% , significantly higher than those of the control group (P <0.05) , and the DU145 cells showed an even higher sensitivity to bortezomib than the LNCaP cells. Bortezomib failed to sensitize these two cell lines to NK cell-mediated cytotoxicity in short-term assay, while long-term assay manifested that the apoptosis rates of DU145 and LNCaP cells after treated with 20 nmol/L bortezomib + NK cells were (41.83 +/- 5.06)% and (30.31 +/- 3.62)% , respectively, significantly higher

CONCLUSIONBortezomib enhances the sensitivity of than those after treated with either bortezomib or NK cells alone (P <0.05). PCa cells to NK cell-mediated cytotoxicity and adds to the effect of current cancer therapies, and it is more efficacious for androgen-independent prostate cancer.

Apoptosis ; drug effects ; Boronic Acids ; pharmacology ; Bortezomib ; Cell Line, Tumor ; Cytotoxicity, Immunologic ; drug effects ; Humans ; Killer Cells, Natural ; drug effects ; Male ; Prostatic Neoplasms ; pathology ; Pyrazines ; pharmacology

Objective To explore the clinical efficacy and the influence of one-port,three-port and four-port laparoscopic cholecystectomy on quality of life in patients with laparoscopic cholecystectomy.Methods The clinical data of 60 patients with laparoscopic cholecystectomy were reviewed,and to compare the general condition,the influence of postoperative recovery and complications,disease of digestive system and the quality of life index in patients treated by one-port laparoscopic cholecystectomy(one-port group),three-port laparoscopic cholecystectomy(three-port group) and four-port laparoscopic cholecystectomy(four-port group).Results Although the time of operation in oneport group was longer than the other two groups (t =3.82,P ＜ 0.05 ; t =4.04,P ＜ 0.05),there were significant differ ences in the volume of the inoperative blood loss,hospital stay time,hospital costs in one-port group compared with the other two groups (P ＜ 0.05).There was no difference among the three groups in the incidence of postoperative complications (P ＞ 0.05).And when leaving hospital,the score of all dimensionalitis in the scale of Gastrointestinal quality of life index (CIQLI),containing the state of physiological function,state of daily life and social activities,psychological emotional state and quality of life of the total score were all higher than those in the other two groups (t =2.22,1.87,2.31,2.43,all P ＜ 0.05),and there was no difference between three-port group and four-port group(P ＞ 0.05).Conclusion Compared with the three-port laparoscopic cholecystectomy and four-port laparoscopic cholecystectomy,one-port laparoscopic cholecystectomy can reduce the inoperative blood loss,shorten hospitalization time and reduce their fee in hospital,and it also can effectively improve patients'quality of life,restore the patients'normal work and life in the shortest possible time.

Objective To examine the circulating glucose and body weight in the transgenic MKR mouse model who expressed dominant-negative IGF-1 receptor and insulin receptor in skeletal muscle leading to systemic insulin resistance and diabetes. Methods MKR mice were genotyped by PCR analysis of tail DNA.And in these mice we examined the circulating glucose and body weight once a week from 1 to 16 weeks of age, and the circulating insulin and glucose tolerance at age of two-month-old by using C57 mice as controls. Results The descendents of MKR mice kept hereditary feature. And these mice had hyperglycaemia from 3 weeks of age,and an increasing body weight slowly(P

[Objective] To study the inhibitory effect of bcl-2 antisense oligonucleotide(ASODN)combined with radiation on lung carcinoma cells NCI-H446.[Method]The cultured NCI-H446 lung carcinoma cells were divided into 5 groups:control,pure radiation,nonsense+radiation,lipofectin+radiation,ASODN+radiation.Every group except for control were assayed by MTT on the inhibition rates,24h,48h,72h after 10Gy irradiation respectively.[Results]The inhibitions of lung carcinoma cells NCI-H446 were observed in ASODN+radiation,lipofectin+radiation,nonsense+radiation and pure radiation groups.The differences between the ASODN+radiation group and the other three groups were significant(P

The significance and method of green chemistry teaching are expounded in detail during the course of medical chemistry experiment,and a new experiment system of green chemistry suiting with medicine is designed in the paper.

97 cases of jaw bone fractures were treated with rigid internal fixation.16 cases had malocclusion postoperation.The etiology was analyzed and the prevention methods were proposed.Intermaxillary elastic traction or fixation and craniomaxillofacial fixation or craniomentum elastic traction were performed to treat the malocclusion.By this way,14 cases were cured.1 case was operated again to resume the normal occlusion,1 case received occlusal adjustment.Incomplete reduction,incorrect operative performance,deficiency of postoperative intermaxillary fixation and incorrect intermaxillary fixation were the main causes to malocclusion.So anatomic in time reduction,correct operative performance,intermaxillary fixation,application of craniomaxillofacial fixation or craniomentum elastic traction instead of intermaxillary fixation for some special cases are effective methods to correct postoperative malocclusion.

Objective To establish the rapid purification of Coxsackievirus A16 using ultracentrifugation .And To prepare and i‐dentify the neutralizing monoclonal antibody against CA16 .Methods The CA16 culture supernatant was harvested and then con‐centrated by 100K capsule .The concentration of CA16 was purified by cesium chloride ultracentrifugation .Purification of CA16 were identified by transmission electron microscopy .BALB/c mice were immunized with inactivated CA16 .Spleen cells were harves‐ted and fused with SP2/0 myeloma cells ,hybridoma cell strain secreting mAb against CA16 were objected to screening .Character‐ization of the prepared mAb were analyzed by ELISA and microneutralization assay .Results The purified CA16 method of cesium chloride gradient ultracentrifugation was established ,TEM analysis was showed that CA16 particles have icosahedral structure ,the diameters of the viral particles were approximately 20-30 nm .Two hybridoma cell strains secreting mAb against CA16 were ob‐tained ,the subtypes of two mAbs were IgG2a ,the binding titers of Anti/CA16/5 and Anti/CA16/10 were 103 and 104 respectively . Neutralizing titer of the two mAbs were 1∶256 and 1∶1 024 respectively .Conclusion Establishment method of cesium chloride gradient ultracentrifugation was performed to purify CA16 ,the two mAbs with neutralizing ability to against CA16 may become ap‐plication of treatment and vaccine .

Objective To study whether ultrasound combined with microbubbles induces suppress invasion in androgen-independent prostate cancer cells PC-3 and to identify the probable mechanism.Methods Ultrasound with a frequency of 21 kHz and intensity of 46 mW/cm2 in continuous wave mode was used.Ultrasound combined with microbubbles (200 μl SonoVue) was used to treat androgen-independent human prostate cancer PC-3 cells for 30 seconds.PC-3 cells were divided into three groups:control group,ultrasound group and ultrasound combined with microbubbles group.Twelve hours after treatment,cell growth curve we drawn,and transwell chamber model was used to do cell invasion experiments in vitro.Real-time PCR was used to measure the expression matrix metalloproteinase-2 and matrix metalloproteinase-9 messenger ribonucleic acid.Results Twelve hours after treatment,cell growth was not significant difference among three groups (P ＞ 0.05).Twelve hours after treatment,ultrasound combined with microbubbles could significantly inhibit the invasion of human prostate cancer cells (P ＜0.05).Treatment with ultrasound combined with microbubbles down-regulated the expression of matrix metalloproteinase-2 and matrix metalloproteinase-9 messenger ribonucleic acid.Conclusions Ultrasound combined with microbubbles inhibited the invasion in human androgen-independent prostate cancer cells through down-regulation of matrix metalloproteinase-2 and matrix metalloproteinase-9.

Objective To investigate the methods of X-ray diagnosis of various displacement of unstable pelvic fracture in three-dimensional space and its instructive significance in closed reduction.Methods A normal adult pelvic specimen was selected and fixed in a wood-frame at supine position after soft tissue rejection and ligament preservation.With the breakage at the unilateral anterior-posterior ring,models of hemipelvic rotation in the transverse and sagittal planes and hemipelvic vertically upward displacement were induced.Anteroposterior radiographs of the pelvic specimen were made and picture archiving and communication system (PACS) was used to measure width of iliac wing,suprainferior diameter of hemipelvis,vertical displacement of iliac crest,acetabulum roof,pubic tubercle and sciatic tuber and area of obturator foramen.Methods of X-ray diagnosis of various displacements of hemipelvis were concluded and applied in treatment of 43 patients with unstable pelvic fractures.Operation time and intraoperative blood loss were recorded.Postoperative images were evaluated by Matta standard.Results Hemipelvic rotation in transverse plane included eversion and inversion.Width of iliac crest was enlarged and area of obturator foramen was shrunk while extroversion ; on the contrary,an opposite result was observed while inversion.Sagittal rotation included pronation and supination.Pubic tubercle had obvious downward shift,iliac crest presented no change or slight upward shift,suprainferior diameter of hemipelvis was lengthened,iliac roof and sciatic tuber remained their position unchanged and area of obturator foramen was shrunk while pronation; on the contrary,pubic tubercle had obvious upward shift,iliac crest presented no change or slight downward shift,suprainferior diameter of hemipelvis was shortened,iliac roof and sciatic tuber remained their position unchanged and area of obturator foramen was enlarged while supination; iliac crest,acetabular roof,pubic tubercle and sciatic tuber presented equidistant upward shift while the hemipelvis displaced upward vertically.Average operation time was 55 minutes (range,15-85 minutes) and intraoperative blood loss was 26 ml (range,10-50 ml).According to Matta standard,pelvic radiography evaluation at postoperative 3 days was excellent in 31 cases and good in 12 cases,with excellent and good rate of 100％.Conclusion Pelvis X-ray films are able to diagnose various three-dimensional displacement of unstable pelvic fractures and guide closed reduction timely and rapidly to achieve satisfactory result.

Objective To study the effect of interleukin 2(IL-2)of clinical dose range,on the proliferation of human peripheral blood T cells,with special emphasis on the number and functional phenotype of γδT cells.Methods Human peripheral blood mononuelear cells(PBMCs)were isolated by density gradient centrifugation and cultured for 2 weeks at different IL-2 concentrations.Ratio and phenotype of different T cell subpopulations before and after in vitro expansion were explored by immunofluorescence staining.Cell number was estimated by trypan blue staining and cell counting.Results Within the four functional phenotypes of Vδ1 as well as Vδ2 γδT cells,CD27~+cells(including CD27~+CD45RA~+and CD27~+CD45RA~-subsets)expressed lymphoid tissue homing receptor CCR7,whereas CD27~-cells(including CD27~-CD45RA~+and CD27~-CD45RA~-subsets)had the peripheral tissue homing potential.All the studied γδT functional subsets showed the expression of activity related receptors,and the ability of a rapid production of various amount of cytotoxicity related effectors following mitogen stimulation.Although IL-2 at high concentration suppressed the proliferation of CD4 T cells,it may promote the proliferation of γδT cells.The proliferated γδT cells were mainly CD27~-CD45RA~-effector cells.Conclusion IL-2 of the clinical dose range may promote proliferation of human peripheral blood γδT cells,which might have important biological significance in IL-2 based anti-tumor therapy.