BACKGROUND:Subcutaneous fat of human body is a rich reservoir of adipose derived stromal stem cells(ADSCs).ADSCs can proliferate rapidly when being cultured in vitro,and has the capacity of multi-directional differentiation.ADSCs attracted much attention in research of tissue engineered seed cells.OBJECTIVE:To isolate and culture stromal vascular fraction(SVF)cells from rabbit subcutaneous fat in vitro,and to testify whether it has multiple differentiation capacity.METHODS:SVF cells were isolated in vitro from rabbits,and cultured under standard condition.Cellular surface antigens CD44,CD45 and CD29 of passage 3 SVF cells were examined using flow cytometry.Passage 3 SVF calls were induced to differentiate into osteoblasts,chondrocytes,and lipocytes.Oil red staining was used to examine lipocyte induction.Alkaline phosphatase(ALP)staining,alizarin red staining and von Kossa staining were used to examine osteoblast induction.Type Ⅱ collagen immunohistochemical staining and type Ⅱ collagen mRNA RT-PCR were used to examine chondrocyte differentiation.RESULTS AND CONCLUSION:Primary SVF cells were multi-angular or short spindle-shaped.Passage 3 SVF calls were long spindle-shaped.Flow cytometry showed CD44+,CD29+,CD45.Oil red staining exhibited positive reaction in lipocyte induction group.ALP staining,alizarin red staining and Von kossa staining demonstrated positive reactions in osteoblast induction group.Type Ⅱ Collagen immunohistochemical staining and alcian blue staining have suggested positive reactions at 14 days of chondrogenic induction group.RT-PCR of type Ⅱ collagen mRNA test showed that the product band had strong signal at 14 days of chondrogenic induction group compared with that before induction.Above mentioned results have indicated that SVF cells isolated from rabbit subcutaneous fat have identical surface makers of stem cells,and have the ability to differentiate into lipocytes,ostsoblasts and chondrocytes in vitro by induction,and it could be concluded that the SVF cells were ADSCs.

BACKGROUND: Small intestinal submucosa (SIS) has good compatibility with cells and tissues, and has good degradabUity. It is an ideal scaffold for tissue engineering. Inducing adipose derived mesenchymal stem cells (ADSCs) seeded on SIS can construct target tissues, which has the potential to be used in clinical treatment.OBJECTIVE: To prepare decellularized porcina SIS matrix, and testify its biocompatibility with rabbit ADSCs cultured in vitro. METHODS: SIS was processed by enzyme digestion-hypertonic saline decellularization, lyophilized at low temperature, and sterilized by gamma radiation. Paraffin sections were used to observe the effect of decellularization of SIS, and the surface structures of SIS were observed by scanning electron microscope (SEM). Rabbit ADSCs were isolated and cultured, and passage 3 ADSCs were seeded onto one side or both sides of SIS. After one weak of co-culture, the cell-scaffold composites were observed.RESULTS AND CONCLUSION: SIS was white and semi-transparent film. Paraffin sections showed no cells on SIS matrix; electron microscopy showed loose weave structure of serosal surface and dense packing structure of mucosal surface. After one week of co-cultivation, plenty of ADSCs were observed on the surface of SIS. In ADSCs seeded onto one side of SIS group, a large number of cells grew on the superior surface, and few even no cells were observed on inferior surface of SIS. When ADSCs were seeded onto both sides of SIS, cells adhered to SIS in paraffin sections. Results show that enzyme digestion-hypartonic saline decellulariation can decellularize SIS completely, and SIS can support ADSCs growth.

Objective To explore the impact of blockade of SDF-1/CXCR4 signaling pathway by T140 on degradation of typeⅡ collagen in human articular cartilage and to define the mechanism of action of T140. Methods 144 pieces of cartilage (Mankin score of 0 or 1) were obtained from osteoarthritis patients undergoing total knee replacement ( OA cartilage group) and 144 pieces of cartilage (Mankin score of 0 or 1) were obtained from patients receiving traumatic amputation (normal cartilage group). Each group was treated with SDF-1 of 100 ng/mL, then divided into three subgroups A, B, and C. The cartilage tissue in each group was cultured in the nutrient solution containing of T140, MAB310, or SDF-1 (1 000 nmol/L) for 48 or 96 hours. RT-PCR was used to detect expression of typeⅡcollagen mRNA in the cartilage tissue. Results Level of type Ⅱcollagen mRNA was markedly higher in subgroup A than in subgroup B and subgroup C at the same group and the same time (P <0.05). The expression level of type Ⅱcollagen mRNA at the same time and in the same subgroup of OA cartilage group were lower than that in normal cartilage group (P < 0.05). Conclusions SDF-1 induces degradation of typeⅡcollagen in human articular cartilage thruogh the SDF-1/CXCR4 signaling pathway. T140 can block the SDF-1/CXCR4 signaling pathway and reduce the degradation of type II collagen.

AIM:To establish a method for evaluating the principle components of Herba Artemisiae Annuae from different habitats.METHODS:Using Fourier transform infrared spectrometry(FTIR),the characteristic peaks of fingerprints of samples from 12 habitats were recognized and compared.RESULTS:The samples from 12 habitats were obviourly different in number,frequency and intensity of peaks.CONCLUSION:FTIR is applied to analysis of principle components of Herba Artemisiae Annuae from different habitats and an operable method of quality control and discrimination for them is provided.

OBJECTIVE:To observe the efficacy of small dose of uremic clearance granule combined with colon enema on aged patients with chronic renal failure(CRF) . METHODS:60 aged CRF patients were randomly divided into treatment group(n=30) and control group(n=30) . Two groups were given essential treatment. Control group were treated with medicinal carbon additionally. Treatment group were additionally treated with uremic clearance granule(p.o.) combined with colon enema. The course of treatment was 15 days and curative effects were compared between two groups after 30 days of treatment. RESULTS:30 days later clinical symptom of patients in treatment group was obviously improved and the levels of blood urea nitrogen,serum creatinine,blood uric acid and serum potassium were lower than before treatment with significant difference. CONCLUSION:Small dose of uremic clearance granule combined with colon enema show satisfactory effect on CRF in aged patients as well as simple,practical and good compliance. This therapy is worth of spreading and putting into application.

The research in bone tissue engineering is abundant and its development is rapid,however,there has been no ideal scaffold materials.We review the recent articles on bone tissue engineering,including ceramics materials,polymerized materials,materials deriver from natural biological organism and their compound materials

BACKGROUND: Studies demonstrated that small intestinal submucosa (SIS) had no immunogenicity, which can not lead to rejection following transplantation, thus, this is an ideal skin substitutes for natural skin.OBJECTIVE: Basic fibroblast growth factor (bFGF) gene was transfected into bone marrow mesenchymal stem cells (BMSCs),and combined the cells with SIS to construct tissue-engineered skin.METHODS: BMSCs were obtained from Japanese big-earad rabbits, and in vitro cultured. Then the subculturad BMSCs were transfected by pCDNA3.1 plasmid, followed by incubation on swine SIS to construct the tissue-engineered skin. The growth of cells and phenotype of BMSCs were detected by flow cytometry. In addition, the result of transfecting BMSCs with pCDNA-bFGF vector was measured by Western blot, and the structure of tissue-engineered skin was observed.RESULTS AND CONCLUSION: After passaged, BMSCs were grown quickly with Iong-fusiform shape. The cells were positive expressed CD90 and CD44, but negative expressed CD45. bFGF had been transfected into BMSCs, and stable expressed. The transfected BMSCs grew well in SIS. By this method, tissue-engineered skin can be constructed in vitro.

Objective To evaluate the efficacy of living donor liver transplantation in the treatment of infants with end-stage liver diseases. Methods The clinical data of 33 infants who received living donor liver transplantation at the Renji Hospital of Shanghai Jiaotong University from October 2006 to September 2009 were retrospectively analyzed. The median age of the infants was 10.9 months, and the mean body weight was 8.2 kg.All of the grafts were left lateral lobes. Tacrolimus (or cyclosporine A) + steroid or tacrolimus (or cyclosporine A)+ steroid + mycophenolate mofeti] were applied to the infants to suppress the immune reaction. Operative techniques, perioperative management and results of follow-up were analyzed. Results The mean operation time,blood loss and blood transfusion of the donors were (384±108)minutes, (183±35) ml and O, and the three indexes of the recipients were (500± 103) minutes, (296±163) ml and (292 ± 159) ml , respectively. The cold preservation time of the grafts was (64 ±23)minutes, the mean weight of the grafts was (249 ±52)g, and the mean graft to recipient weight ratio was 2.1% ± 0.4%. All donors recovered smoothly and no complication occurred. Of the recipients, three were complicated with hepatic artery thrombosis, two with portal vein thrombosis,nine with biliary complications, 11 with infection, two with acute rejection and five infants died perioperatively.The one-year cumulative survival rate of the infants was 85% (28/33). Conclusions Infants with end-stage liver diseases could be treated by living donor liver transplantation. The development of surgical techniques and perioperative managements improves the success rate of operation and the long-term survival rate.

Objective To assess the symptoms of patients due to indwelling double ureteral stent and their impact on quality of life. Methods Sixty patients were assessed by the specific questionnaire. Average time of indwelling stent was 21.4 d. All patients finished the special questionnaire when the stent was removed by cystoscope, including specific urinary symptoms, IPSS (international prostate symptom score), VAS (visual analogic scale) and QOL (quality of life). To assess the pain feeling, the patients were divided into 3 groups by varied methods of anesthesia, including local anesthesia, balance anesthesia and general anesthesia. Results Insertion or removal of ureteral stents with local anesthesia provoked pain in 11 of 12 (91%) patients and 8 of 10 (80%) patients in balance anesthesia group. But 38 patients of general anesthesia group did not feel any pain when inserting stents. In 54 of 60 (90%) patients, the indwelling catheter provoked one or several urinary symptoms: nocturia (70%), frequency (65%), urgency (60%), tenesmus (58%), dysuria (52%), hematuria (35 % ) and incontinence (30 %). 75 % of the patients experienced pain, in the flank and lower abdomen. 29 (48 %) patients were found to be unsatisfied with their quality of life due to the indwelling stent. Among them, 18 (62%) patients demonstrated that the provoked overactive bladder symptoms were the main influence factor of QOL, compared with 5(17 %) patients complaining pain feeling. Conclusions Urinary symptoms and pain associated with indwelling double J ureteral stents could interfere the daily activities and 50 % of patients were found reduced quality of life. Overactive bladder symptoms were the main influence of QOL during the time of indwelling stent.

Objective To compare T and N staging of esophageal and cardia cancer by endoscopic ultrasonography (EUS) and computed tomography (CT) before operation, and to evaluate clinical value of EUS.Methods Twenty-eight patients received examination of EUS and CT preoperatively, and the T and N staging was determined. The accuracy rates of both T, N staging were compared by the postoperative pathological results, at the same time, the accuracy rates of lymph node metastasis were compared, and the value of application of EUS was investigated. Results In the twenty-eight cases, the accuracy rates of EUS were higher than that of CT by 89.3% ( 25/28 ) to 46.4% ( 13/28 ) in T staging. And the difference had statistical significance( P = 0. 004 ,P ＜ 0.01 ). The N stagings of both were 82.1% (23/28) and 50.0% ( 14/28), and the difference had statistical significance ( P = 0. 035, P ＜ 0. 05 ). The accuracy rates of lymph node metastasis of both were 88.7% and 72.2% respectively, and the difference had statistical significance (χ2 = 7.031,P = 0.008, P ＜ 0. 01 ). Conclusions EUS has very important value in preoperative staging for esophageal cancer and cardia cancer, and the T staging of EUS is better than that of CT. The short axis and the ratio of short to long axis (S/L) combined with the lymph node ultrasonography image can improve the accuracy rates of lymph node metatsasis and N staging.