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 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

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.

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.

Autophagy is an important homeostatic cellular recycling mechanism responsible for degrading injured or dysfunctional cellular organelles and proteins in all living cells. Aging is a universal phenomenon characterized by progressive deterioration of cells and organs due to accumulation of macromolecular and organelle damage. Growing evidences indicate that the rate of autophagosome formation and maturation and the efficiency of autophagosome/lysosome fusion decline with age. Dysfunctional autophagy has also been observed in age-related diseases. Autophagy disruption resulted accumulation of mutated or misfolded proteins is the essential feature of neurodegenerative disorders. However, in cancers, fibroproliferative diseases or cardiovascular diseases, autophagy can play either a protective or destructive role in different types of disease, and even in different stages of the same disease. The review will discuss the cellular and molecular mechanisms of autophagy and its important role in the pathogenesis of aging and age-related diseases, and the ongoing drug discovery strategies for therapeutic intervention.
Aging ; Autophagy ; Drug Discovery ; Humans ; Lysosomes ; metabolism ; Neurodegenerative Diseases ; Phagosomes ; metabolism ; Protein Folding

Objective To establish an expression system of TCR?9/?2-Fc protein by baculovirus vector expression system and identify biological function of expressed TCR?9/?2-Fc protein.Methods ?9Fc and ?2(OT3)Fc gene fragments were amplified by overlap PCR and inserted into expression vector pBACp10ph.The recombinant plasmid pBACp10ph-?9/?2(OT3)-Fc and the baculovirus DNA were co-transfected into sf9 cells.The expression position of TCR?9/?2(OT3)-Fc was identified by Western blot and the expression efficiency of ?9Fc and ?2(OT3)Fc was tested by flow cytometry(FCM).Furthermore,the binding activity of TCR?9/?2(OT3)-Fc protein with SKOV3 cells and MNS protein was evaluated with laser scanning confocal microslopy and surface plasmon resonance(SPR).Results The recombinant vector pBACp10ph-?9/?2(OT3)-Fc was constructed and TCR?9/?2(OT3)-Fc protein was expressed in sf9 cells.However,the expression efficiency of ?9Fc and ?2(OT3)Fc was quite different.It was proved that purified TCR?9/?2(OT3)-Fc protein can bind with SKOV3 cell and MNS protein.Conclusion TCR?9/?2-Fc protein is successfully expressed in baculovirus vector expression system and TCR?9/?2-Fc protein can simulate the binding activity of TCR in vitro.

Objective To detect amniotic fluid ABH blood group substances and ABO blood group genotype by the polymerase chain reaction with sequence-specific primers(PCR-SSP) to increase the prenatal diagnosis of fetal ABO blood group .Methods 53 pregnant women with gestational age 16 -25 weeks were selected .Amniotic fluid was extracted for detecting ABH blood group substances by the serological indirect agglutinating reaction ;the amniotic fluid cells were separated for extracting DNA .Then the PCR-SSP technique was adopted to analyze the ABO blood group genotypes .Results 16 specimens of amniotic fluid were non-se-creting type phenotype(30 .2% ) and 37 specimens of amniotic fluid were secreting type phenotype (69 .8% );48 specimens of amni-otic fluid were detected out the ABO blood group genotype by the PCR-SSP method .ABO blood group of fetal amniotic fluid cells by the gene identification was consistent to the detection results of amniotic fluid secreting type ABH blood group substances .Con-clusion The PCR-SSP technique can accurately detect the fetal amniotic fluid cells ABO blood group .

Objective To evaluate the effect of combination of surgery and transcatheter hepatic arterial chemotherapic embolism(THACE) with or without placement of drug deliery system(DDS) of portal vein for hepatocellular carcinoma with portal vein tumor thrombus (PVTT). Methods Fifteen cases of advanced HCC with PVTT were treated by hepatectomy, thrombectomy of portal vein with THACE,and 5 of the 15 cases received postoperative chemotherapy via portal vein. Results All patients survived after operation. The survival rate of 6,12,18 months was 15,12 and 9 cases respectively. Conclusions Operation is still an effective method for advanced HCC , postoperative comprehensive treatment can improve the survival rate.

BACKGROUND: Mesenchymal stem cells are few in human bone marrow, and their number will decrease with aging or body weakening, so a large amount of amplification is necessary. However, the biological characteristics of human mesenchymal stem cells of each passage remain poorly understood.OBJECTIVE: To analyze and compare the biological characteristics of each passage of bone marrow-derived mesenchymal stem cells (MSCs) so as to provide a basis for clinical demands of tissue engineering.DESIGN,TIME AND SETTING: Cytological observation in vitro. The experiment was performed at the Department of Orthopedics and Central Laboratory, Dongzhimen Hospital, Beijing University of Chinese Medicine from March to October 2008.MATERIALS: From bone marrow of patients with non-hematopoietic disease, MSCs were provided by Department of Orthopedics, Dongzhimen Hospital, Beijing University of Chinese Medicine.METHODS: Bone marrow was collected form posterior superior iliac spine of patient, MSCs were isolated and cultured by Percoll method. When the cells were confluent at 90%, they were trypsinized and observed by inverted miscroscopy. The second passage of cells were collected for index detection.MAIN OUTCOME MEASURES: Cell morphological characteristics and immunophenotype; cell activity was detected by MTT; cell division and apoptosis in the proportion of necrosis were analyzed by flow cytometry analysis.RESULTS: The passaged MSCs exhibited uniform appearance in fusiform shape, and their growth was slowed down after 9 passages, exhibiting cytoplasm vacuolization and body enlarging. The second passage of MSCs was positive for CD44, CD106,and CD105, but negative for CD34 and CD45. MTT values peaked at passage 9, and gradually decreased since passage 10. At passage 11, the number of MSCs at division stage was increased, but from the sixth passage, the number of apoptotic cells increased significantly, reaching more than 60% at passage 8.CONCLUSION: According to biological characteristics analysis of MSCs at each passage, the third to the sixth passage cells are recommend for clinical therapy.