BACKGROUND: Adipose tissue-derived stem cells receive a high attention in tissue engineering research. Adipose tissue-derived stem cells lack of specific surface marker, there is no effective purified method. Purified adipose is a simple method to elevate purify of stem cells. OBJECTIVE:To analyze how to purify adipose tissues before primary culture of adipose tissue-derived stem cells. DESIGN, TIME AND SETTING: The controlled animal experiment was performed at the Shanghai Animal Center of Experimental Medicine of Shanghai Sixth People’s Hospital between December 2007 and March 2008. MATERIALS: Four-week old Sprague Dawley rats were used for obtaining adipose tissues from the inguinal groove. METHODS: Adipose tissues from rat inguinal groove were dissected to educe superficial blood vessel and blood vessel branches. Both blood vessel inside and elliptic nodal tissues surrounding blood vessels were excised. MAIN OUTCOME MEASURES: Stained elliptic nodal tissues stained by Hematoxylin-Eosin were observed with a microscope to make sure what kind of tissues they are. The purified adipose tissues and unpurified adipose tissues were stained by Hematoxylin-Eosin. The differences in their tissue construction were observed using the microscope. RESULTS: Elliptic nodal tissues stained by Hematoxylin-Eosin were proved to be lymphatic tissues. The tissue construction of purified adipose tissues was pure, and the cellular component was simple. Conversely, the tissue construction of unpurified adipose tissues was complicated, and cells were various with complicated components. CONCLUSION: The component of adipose tissues used to primary cultured adipose tissue-derived stem cells is complicated. As resection of superficial blood vessel, skin and muscle tissues, blood vessel inside tissues and lymphatic tissues should also be excised.

OBJECTIVETo analyze the efficacy and medication principles of Professor Xu Fu-songs traditional Chinese medicine (TCM) protocols for male diseases.

METHODSWe reviewed and descriptively analyzed the unpublished complete medical records of 100 male cases treated by Professor Xu Fu-song with his TCM protocols from 1978 to 1992.

RESULTSThe 100 cases involved 32 male diseases, most of which were difficult and complicated cases. The drug compliance was 95%. Each prescription was made up of 14 traditional Chinese drugs on average. The cure rate was 32% , and the effective rate was 85%. Professor Xu Fu-song advanced and proved some new theories and therapeutic methods.

CONCLUSIONProfessor Xu Fu-song's TCM protocols can be applied to a wide range of male diseases, mostly complicated, and are characterized by accurate differentiation of symptoms and signs, high drug compliance, and excellent therapeutic efficacy.

ObjectiveTo assess the feasibility of seeding adipose-derived stem cells (ADSCs) onto bladder acellular matrix grafts (BAMGs) for bladder reconstruction in a rabbit model.MethodsAutologous ADSCs were isolated,expanded and identified by flow cytometry.In the experimental group,ADSCs were seeded onto BAMGS for reconstructing bladder defects in 12 male rabbits.Unseeded BAMGs were used for bladder reconstruction in the control group of 12 rabbits.Cystography was performed at 24 weeks after grafts implantation.Following cystography,the animals were scarified and grafts were harvested； H&E and immunohistochemical staining were performed with cytokeratin AE1/AE3,smooth muscle α-actin and S-100 markers.ResultsFlow cytometry demonstrated that the ADSCs expressed CD90,CD44,CD105,CD166 and CD34,but not CD45 or CD106.The cells demonstrated good biocompatibility with BAMGs.At 24 weeks,in the experimental group,the reconstructed bladders reached a mean volume of (94.68 ± 3.31 )％ of the precystectomy bladder capacity.Complete regeneration of smooth muscle and nerve tissue was evident.Regenerated SMCs,urothelium and nerve cells stained positively for α-smooth muscle actin,AE1/AE3 and S100.In the control group,the mean bladder volume was (69.33 ± 5.05 )％ of the pre-cystectomy volume.Histologically,the control group was characterized by multi-layered urothelium without evidence for organized muscle or nerve tissue.Conclusion The tissue engineering bladder constructed by ADSCs and BAMG can be used as an ideal biomaterial to replace and repair the bladder.

Stress urinary incontinence is one of the most common diseases in urinary system.At present,the major methods for treating stress urinary incontinence include medication,physico-behavior therapy and operation.However,for various reasons,the current methods do not yield satisfactory results.As a newly emerging technique,tissue engineering provides a new concept and method to treat stress urinary incontinence.The application of tissue engineering in the treatment of stress urinary incontinence is reviewed in this article.

Objective To explore the method of culture of rat adipose-derived stem cells(ADSCs) and differentiation induction into myoblasts. Methods Adipose tissues were obtained from SD rats,and were isolated by enzyme digestion and cultured into ADSCs.The expression of surface antigen CD90,CD105 and CD34 was detected by immunofluorescence and flow cytometry.ADSCs of the second passage with logarithmic growth were obtained,and culture media containing 5-azacytidine(5-aza) and basic culture media were employed for cells in induction group and control group,respectively.The induction lasted for 7 d,14 d,21 d,28 d and 35 d,respectively.Cell growth and cell morphology were observed by inverted phase contrast microscope,and immunofluorescence and flow cytometry were utilized to detect the expression of myoblast specific antigens desmin and myosin. Results ADSCs were successfully isolated and cultured,and were identified to be stem cells.On the 28th day of induction,cells in induction group displayed "swirl" morpholgy,and multinucleation was observed.It was revealed by immunofluorescence and flow cytometry that the highest expression rates of desmin and myosin were 52.57% and 50.04%,respectively on the 28th day of induction,while there was no expression before induction and in control group. Conclusion ADSCs can be isolated and cultured from rat adipose tissues,and can further differentiate into myoblasts after induction by culture media with 5-aza.The expression of myoblast specific antigen is the highest on the 28th day of induction.

Objective To analyze the clinical characters,treatment effects and influential factors of gastrointestinal stromal tumor patients with hepatic metastases,and to explore the reasonable clinical treatment protocols.Methods The clinical data of 24 gastrointestinal stromal tumor patients with hepatic metastases who were admitted to Peoples Liberation Army Hospital from April 2007 to April 2010 were retrospectively analyzed.Results The 1-,2-,3-and 4-year cumulative survival rates of hepatic metastases patients with gastrointestinal stromal tumors were 91％,72 ％,22 ％ and 6 ％,respectively.The 1-,2-and 3-year survival rates of the patients treated with imatinib after operation were 91％,70 ％,28 ％,respectively,the median survival time was 26 months,the mortality rate was 27.3 ％ (3/11),and 9 cases got the tumor complete elimination.The 1-,2-and 3-year survival rates of the patients treated without imatinib after operation were 90 ％,68 ％,18 ％,respectively,the median survival time was 24 months,the mortality rate was 14.3 ％ (2/13),and 12 cases got the tumor complete elimination.The recurrence rate after complete resection was 28.6 ％(4/14),and the mortality rate was 7.1％ (1/14).The mortality rate after incomplete resection was 40 ％ (4/10).Conclusion Surgical treatment of eliminating the primary and metastatic lesion,and adjuvant therapy of imtinib could prolong the survival time,reduce postoperative recurrence rate and improve the quality of life,which are the effective methods for the treatment of the gastrointestinal stromal tumor patients with hepatic metastases.

Objective To evaluate the clinical application of the fistulography of multi-slice spiral CT(MSCT)in the diagnosis of anal fistula.Methods A total of 28 patients who were verified operatively of fistula in ano underwent preoperative fistulography of MSCT.The multi-planar reformation(MPR)and surface shadow display(SSD)images were generated and analyzed retrorespectively.Results There were 8 cases with perianal abscess and 20 cases with anal fistula in 28 patients.Complex fistula was diagnosed in 16 cases,and simple inter-sphincteric fistula was found in 4 cases;the inner orificiums of anal fistulas were revealed accurately with MPR images in 18 cases.Conclusion The fistulography of MSCT is valuable for preoperative assessment of anal fistula,in particular for investigation of the inner orificiums of anal fistulas with MPR.

Objective:To express and purify recombinant human collagen type Ⅲ and evaluate its properties.Methods:The recombinant genetic engineering strain pET30a(+)-1880/pACYCDuet-hy726/bL21(DE3) was constructed to stably co-express recombinant human type Ⅲ collagen (rhCol) and prolyl hydroxylase. rhCol was prepared and purified by E. coli high-density fermentation, salting out and column chromatography protein purification technology. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to determine the purity of rhCol. The N-terminal amino acid sequences of rhCol were determined by automatic protein polypeptide sequencing instrument. The hydroxyproline content of rhCol was determined by ultraviolet spectrophotometry. The cellular compatibility of rhCol was evaluated by MTT assay. Results:The final wet weight of high-density fermentation was about 200 g/L. The expression level was about 3 g/L. The purity of rhCol by affinity chromatography was over 95%. The results showed that the hydroxyproline content of rhCol was 11.44%, and the rhCol products have good water solubility and cell compatibility.Conclusions:RhCol can be widely applied to the field of skin care and biomedicine as an excellent biological material.

Objective To conduct a pilot study on genome-wide in vivo protein-RNA interactions in E.coli.Methods Bacterial lysate was treated with RNase before the RNA fragments protected by proteins were extracted from treated lysate and used to construct cDNA library that was applied to high-throughput sequencing .Finally, the transcripts bound by proteins were obtained by bioinformatics analysis .Results A total of 3193 transcripts were obtained , including 2234 mRNAs, 47 sRNAs, 39 tRNAs, 11 rRNAs, and 862 intergenic regions .Conclusion Some information of transcripts interacting with proteins in E.coli is acquired , which will facilitate further studies of protein-RNA interactions .

Adenocarcinoma ; pathology ; Adenocarcinoma, Papillary ; pathology ; Aged, 80 and over ; Carcinoma, Renal Cell ; pathology ; Duodenal Neoplasms ; pathology ; Humans ; Kidney Neoplasms ; pathology ; Lung Neoplasms ; pathology ; Male ; Neoplasms, Multiple Primary ; pathology ; Sarcoma ; pathology ; Stomach Neoplasms ; pathology