BACKGROUND: Embryonic pancreatic tissue is characterized by its abundance, potent in proliferation & differentiation, and minimal immunological rejection. It is widely considered as potential pancreatic endocrinological stem cells resource for treating diabetes mellitus.OBJECTIVE: To investigate the embryonic mouse pancreatic tissue isolation technique and observe the recipients' blood glucose regulatory effects of the grafted embryonic pancreas in an experimental diabetes mellitus mouse model.METHODS: Pancreatic tissue from C57B1/6 mouse embryos at embryonic days 11.5-16.5 was isolated under the stereomicroscope. C57BL/6 mouse models of streptozocin-induced diabetes mellitus were established and then randomly divided into two groups: transplantation group, in which, five pieces of pancreatic tissue of mice at embryonic 16.5 days were transplanted into mouse renal capsule, and sham-operated control group, in which, 0.05 mL RPMI1640 culture medium was injected into mouse renal capsule. When blood glucose level of the transplantation group mouse was≤ 11.2 mmol/L, the endocrine function of embryonic pancreatic tissue transplanted was detected by IPGTT and IPITT methods and then the transplanted graft was removed for observing the blood glucose relapse.RESULTS AND CONCLUSION: Nearly intact pancreatic tissue of mice at embryonic days 11.5-16.5 could be isolated through the use of stereomicroscope. Pancreatic tissue morphology and color of mice ≤ embryonic 12.5 days were difficultly distinguished from adjacent tissue and they could only be isolated carefully according to the relationship with adjacent organs. Pancreatic tissue of mice > embryonic 12.5 days exhibited initial endocrinological tissue morphology mimic white cauliflower. Histological and ELISA examinations showed that embryonic pancreatic tissue could express and secrete insulin and the insulin level was gradually increased with developmental time. Embryonic pancreatic tissue could grow beneath the recipient renal capsule. The insulin and glucagon expression in the post-transplantational pancreatic tissue graft was increased compared with prior to transplantation. These results suggest that pancreatic tissue is a potential stem cell resource for treating the diabetes mellitus.

0.05).Compared with control group,significant decrease in positive group was found in the interval from first evacuation of HM to resolution of serum ?-hCG level,(83?18) days versus(126?31)days(P0.05).Conclusions Once HM is diagnosed,evacuation should be performed as soon as possible,the later the evacuation begins,the higher the risks of lung metastasis and chemotherapy are.It is not necessary to worry about lung metastasis before evacuation of HM,the outcome of post- chemotherapy is very good.

Objective To discuss the feasibility on building lewis lung carcinoma mouse models through different methods and improve the methods. Methods The method of culture LLC cells in vitro, trypsin digestion method, Ⅳ collagenase method and homogenate method were compared to make the different dose of cell suspension injected into C57BL/6 mice. The feasibility of the improved method was determined through observing the cell count, the tumor formation ratio, the tumor formation time, tumor volume, weight and life habit. Results The method of culture LLC cells in vitro could get needed cells and its tumor formation ratio was 100 %. Trypsin digestion method and homogenate method could get less cells and its tumor formation ratio was about 80 %～90 % and 60 %～75 %. Whereas 1V collagenase method could get most cell count and its tumor formation ratio was 100 %. Conclusion IV collagenase method is a preferred method which is simple,high efficiency and make a strong base on the cancer experimental study.

Objective To detect the efficiency and function of NK cell differentiation from human umbilical cord he?matopoietic stem cells (HSCs) in vitro. Methods CD34+hematopoietic stem cells were isolated from human umbilical cord blood, and inoculated into SCGM medium containing 20 μg/L FMS like tyrosine kinase 3 ligand (Flt-3L), stem cell fac?tor (SCF), interleukin (IL)-7, IL-15 and IL-21. And CD34+HSCs were differentiated into NK cells in directional inducing. The growth state of cells was observed. The expressions of CD56, NKG2D, NKp46, CD3, CD19 and CD34 were detected by flow cytometry in the differentiation of 7, 14, 21 and 28 d. In the differentiation of 21 d and 28 d, the differentiation cells were used as effector cells, and K562 cells as target cells. The ratios of effector cells and target cells were 8∶1, 4∶1, 2∶1 and 1∶1. The killing activity of the differentiated cells was detected by lactate dehydrogenase (LDH) cell toxicity assay and 7AAD/CFSE labeling method. Results CD34+HSCs derived from human umbilical cord blood can proliferate in vitro under appropriate condition. There were no significant differences in the expression of CD3 and CD19 between different differentia?tion stages (7, 14, 21 and 28 d, P>0.05). The expressions of CD56, NKG2D and NKp46 were significantly different (P<0.05), and the ultimate expression amount was (72.57±1.60)%, (32.83±1.29)%and (29.53±2.40)%. The expression of CD34 decreased gradually, and the lowest was (12.13 ± 2.01)%. The maximum killing activity detected by LDH cell toxicity assay and 7AAD/CFSE labeling method reached(49.91±2.76)%and (40.87±1.12)%.The killing activity of NK cells was decreased in the order of 8∶1, 4∶1, 2∶1 and 1∶1 groups (P<0.05). There was no significant difference in the killing activity between NK cells of 28 d and 21 d. Conclusion Human umbilical cord hematopoietic stem cells can differentiate into NK cells un? der appropriate conditions in vitro, and the NK cells induced from differentiation are with killing activity.

OBJECTIVETo explore the effects of stromal interaction molecule 1 (STIM1) on the expression of apoptosis-related proteins in prostate cancer PC-3 cells.

METHODSWe transfected the lentivirus vector STIM1-pGCSIL-GFP carrying STIM shRNA into human hormone-independent prostate cancer PC-3 cells, and 3 days later observed the transfection efficiency by fluorescence microscopy. At 7 days after transfection, we determined the expression of STIM1 in the PC-3 cells by RT-PCR and Western blot and those of apoptosis-related proteins Bcl-2, Bax, survivin and activated Caspase-3 by Western blot.

RESULTSAt 3 days, inverted microscopy revealed a transfection efficiency of > 80%. At 7 days, the STIM1 expression was significantly inhibited at both mRNA and protein levels. The Bcl-2/Bax rate was remarkably decreased as compared with that of the control group (0. 31 vs 1.24 ) , and the survivin expression was markedly reduced, 0. 14 times that of the relative expression in the control. However, the Caspase-3 cleavage was significantly activated, 1.52 times that of the control (P <0.05).

CONCLUSIONSTIM1 can be regarded as an oncogene in prostate cancer PC-3 cells. Inhibition of its expression can induce PC-3 cell apoptosis by reducing the Bcl-2/Bax rate, decreasing the survivin expression, and activating the Caspase-3 pathway.

Apoptosis ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Male ; Membrane Proteins ; genetics ; Neoplasm Proteins ; genetics ; Prostatic Neoplasms ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Small Interfering ; genetics ; Stromal Interaction Molecule 1 ; Transfection ; bcl-2-Associated X Protein ; metabolism

Objective To investigate the significance of resistant gene detection combined with adenosine triphosphate-tumor chemosensitivity assay (ATP-TCA) in the second-line chemotherapy of lung squamous cell cancer, and to provide a reference for clinical treatment. Methods 150 patients with lung squamous cell cancer diagnosed by histopathology or cytology and with the disease progressed after NP regime chemotherapy were enrolled. The mRNA expressions of excision repair cross complementation 1 (ERCC1) and ribonucleotide reductase M1 (RRM1) were detected by RT-PCR, and ATP-TCA was carried out. After detected by RT-PCR and ATP-TCA, the patients who were sensitive to gemcitabine plus cisplatin (GP) accepted the second-line systemic chemotherapy with GP regimen, and the others who were not sensitive to GP regimen or whose results of gene detection and ATP-TCA were on the contrary took the second-line chemotherapy regimens with docetaxel plus cisplatin (DP). Both groups accepted 2-4 cycles of systemic chemotherapy. The chest CT was followed up, and the response rate (RR), progression-free survival (PFS) and median survival time (MST) were investigated. Results The RR of GP group was 36.2 % (17/47), while the DP group was 28.1 % (26/92), and the difference was statistically significant (χ2= 4.274, P< 0.05). The media PFS of GP group and DP group were 4.2 months (95%CI 3.485-5.348 months) and 3.6 months (95 %CI 2.685-4.648 months), respectively, and the difference was statistically significant (P<0.05). The MST of GP group and DP group were 9.6 months (95 %CI 8.283-10.637 months) and 8.9 months (95 %CI 7.384-9.648 months), respectively, and there was no statistically significant difference (P> 0.05). Conclusion The resistance gene detection combined with ATP-TCA have certain guiding significance on the second-line chemotherapy for advanced lung squamous cell cancer.

Ethanol can be produced from lignocellulose by first hydrolysing the material to sugars,including hexose,and pentose,and then fermenting the hydrolysate to ethanol.Hydrolysis using dilute-acid has advantages over other methods.However,compounds which inhibit fermentation are generated during this kind of hydrolysis.Therefore,it is important to focus on microorganisms metabolizing xylose and tolerating/decomposing inhibitors,on detoxification methods of hydroly- sates with low-cost and facilitated to scale-up,and different fermentation modes in ethanol production from hydrolysate.This review summarized the advance in above aspects.

Objective To explore the feasibility of culturing dermal papillae cells (DPC) of hu- man hair in a serum-flee medium,and to observe the growth characteristics of these cells.Methods Cell culture flasks (plates) were pretreated with fibronectin,and DPC (2nd passage) were incubated with Williams E serum-flee medium supplemented with insulin-transferrin-selenite (ITS).Cells were observed by an inverted phase-contrast microscope.Proliferation of DPC was evaluated with 3-(4,5-dimethylthia- zol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and by their growth curve.Results In a serum-free medium,2nd passage DPC adhered to the flask surface within two to four hours of incubation; two to three days later,confluence,of the cells was observed,without noticeable proliferation.Four days later,cell connection was interrupted,isolated cells or cell clusters were seen,and detachment of some cells from the flask surface was observed.One to two weeks later,most cells had died.After incubation with 4% bovine serum for ten hours,cell proliferation was observed surrounding the remaining viable cell colonies. DPC growth curve showed stagnant phase and slow growth phase;however,log growth phase was not ob- served.Conclusion DPC could be successfully cultured in serum-free medium.However,the culture con- dition needs to be further optimized.

Objective To investigate the effect of rehabilitation stroke unit on walking in stroke patients with hemiplegia. Methods 86 patients with walking impairment after stroke were randomly divided into control group (n=43), who accepted conventional treatment, and experimental group (n=43), who were incorporated into the rehabilitation stroke unit. They were assessed with Berg Balance Scale (BBS),10 m Maximum Walking Speed (MWS), Fugl-Meyer Assessment (FMA) and modified Barthel Index (MBI) before and 8 weeks after treatment.Results The scores of BBS, MWS, FMA and MBI improved after treatment in both groups (P<0.01), and improved more in the experimental group than in the control group (P<0.05). Conclusion Rehabilitation stroke unit is effective on walking ability in stroke patients with hemiplegia.

OBJECTIVETo construct and identify the eukaryotic expression plasmids encoding two short hairpin RNA (shRNA) of survivin for the purpose of paving the way for the studies of targeted gene therapy for prostatic carcinoma (PCa).

METHODSTwo shRNA of survivin were designed and synthesized respectively, and then both were cloned into plasmids. Finally, the recombinant plasmids were confirmed by sequencing and agarose gel electrophoresis after restriction digestion.

RESULTSThe recombinant plasmids encoding two survivin shRNA were constructed and the aim sequence obtained.

CONCLUSIONSuccessful construction of the recombinant provides a sound basis for the research of targeted gene therapy for PCa.

Cloning, Molecular ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; Neoplasm Proteins ; biosynthesis ; genetics ; Plasmids ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection