Objective To explore the difference of immunosuppresive effect between the expanded Stro-1+ and Stro-1-subgroups of human mesenchymal stem cells in vitro. Methods Mononuclear cells (MNCs) were isolated from bone marrow (BM) samples and seeded in a T-75 cm2 tissue culture flask contained with Dexter medium. When 50% confluence was obtained, adherent cells were collected and incubated with anti-stro-1 antibody, and the Stro-1+ and Stro-1-MSCs were further seeded for expansion. The total culture time (median) was 15 days. Cells were then analyzed by flow cytometry. One-way mix lymphocyte reaction (MLR) (1?105 responding cells and an equal number of stimulating cells/well were co-cultured in 96-well plates) and nonspecific mitogenic stimuli phytohemagglutinin (PHA) plus interleukin-2 (IL-2) induced lymphocytes proliferation (PBL 1?105/well were mixed with PHA 10?g/ml and IL-2 500U/ml in 96-well culture plates) were established in vitro. 1?103-3?104 irradiated Stro-1+ MSCs and Stro-1-MSCs were added into the two systems at the beginning of reaction. Immunosuppressive actions of both Stro-1+ or Stro-1-MSCs were compared. Results Adherent cells contained a median of 9% (range 5%-26%) Stro-1+ cells, which expressed higher immunophenotype of MSCs. In both reaction systems, suppressive actions occurred in a dose-dependent fashion when whatever Stro-1+ MSC or Stro-1-MSC were added. However, that the addition of 1?103 Stro-1-cells enhanced rather than inhibited the lymphocyte proliferation in one-way MLR. In the presence of various concentrations of MSCs, Stro-1+ MSCs always showed a significantly increased inhibitory effects in comparison to Stro-1-MSCs (P

35), while the expression level of TGF-?1 in Stro-1-MSC was significant higher than that detected in Stro-1+MSC (2-??Ct=0.39, P

Objective: To reconstruct adenovirus vector of breast eukaryotic initiation factor 4E and to observe its effect on the metastasis ability of breast cancer cell line MCF-7. Methods: eIF-4E gene was constructed into adenovirus vector pAD-X by gene recombination technique, which was transformed into 293 packaged cell for high titer adenovirus. Real-time PCR was applied to detect eIF-4E gene expression. eIF-4E siRNA was applied and then transwell cabin assay was used to observe changes of invasion and motor ability of MCF-7 cells transfected with reconstruction adenovirus. Result:The finding of digestion was coincided with expected. eIF-4E gene over-expression was detected in transfected MCF-7 cells with real-time PCR. And the invasion and motor abilities of transfected MCF-7 cells were more significantly inhibited in transwell cabin assay (respectively p

Objective To investigate the expression of PCNA in gastric cancer and its relationship with telomerase activity of peritoneal washings and peritoneal dissemination, and to compare the efficacy of telomerase activity and cytology of peritoneal washings for prediction of peritoneal metastasis of gastric cancer. Methods Telomeric repeated amplification protocol (TRAP)-enzyme-linked immunosorbent assay (ELISA) was performed to measure the telomerase activity of peritoneal washings collected from 60 patients with gastric cancer. Exfoliate cytologic analysis of the corresponding samples was used for comparison.Expression of PCNA was measured with immunohistochemical staining.Their relationship with clinicopathologic features were evaluated. Results The positive rate of telomerase activity in peritoneal washing collected from patients with gastric cancer was 41.7%,which well related to serosal invasion, histology types, depth of infiltration and peritoneal metastasis of gastric cancer. The positive rate of telomerase activity increased with the increased depth of infiltration and serosal involvement areas (P

Background Corneal transplantation is a primary method for the treatment of serious corneal diseases, but its application is limited because of the shortage of corneal donor.The study on tissue engineering corneal epithelium provides a new approach to corneal transplantation, and the biological scaffold materials for tissue engineering corneal epithelium is an issue of increasing concern.Bacterial cellulose membrane has been used in medical field,but its application in tissue engineering corneal epithelium deserves more researching.Objective This study was to evaluate the biocompatibility of bacterial cellulose membrane as a biological scaffold of tissue engineering corneal epithelium.Methods Corneal epithelium was isolated from 1 month-old New Zealand White rabbit.Corneal epithelial cells were cultured using explant method and identified by detecting the CK-3 expression using immunofluorescence technique.The second generation ceils were inoculated on bacterial cellulose membrane and culture plate, respectively, and the growth status of the cells were examined and compared under the optical microscope.The cell activity/toxicity test was performed by LIVE/DEAD cell staining kit at the third day after inoculation to evaluate the survival rate.The ultrastructure of the cell surface was examined under the scanning electron microscope.The study was performed in accordance with the ARVO Statement.Results Rabbit corneal epithelial cells grew well 1 week after primarily cultured with a cobblestone-like appearance and positive response for CK3 antibody.The cells on the bacterial cellulose membrane presented a round shape and regular arrangement and showed the green fluorescence for LIVE/DEAD test,with the survival rate 100％.Abundant leafy protrusion, microvilli and intercellular junction were seen under the scanning electron microscope.In addition, mitosis phase of cells and many filopodia between the cells and bacterial cellulose membrane were also exhibited.Conclusions Rabbit corneal epithelial cells can grow well in bacterial cellulose membrane.Bacterial cellulose membrane has good biocompatibility, indicating that bacterial cellulose membrane can be used as new biological material for tissue engineering corneal epithelium.

Objective To illustrate the expressional distribution of heat shock protein 70(HSP 70) in the kidney of acute renal injury rats caused by gentamicin.Methods One hundred and fifty rats were randomly divided into 3 groups: group A (normal group), group B [80 mg/(kg?d) gentamicin to be injected], group C [160 mg/(kg?d) gentamicin to be given]. The variations of renal pathology were observed at different time phasess by light scope and electromicroscope. Simultaneously, the expression of HSP 70 in kidney was evaluated by immunohistochemistry.Results HSP 70 distributed in epithelial cells of renal tubular in acute injury rats. The expression of HSP 70 increased markedly from the 6th hour after injection, peaked at the 12th hour and lasted for 48 hours. The expression of HSP 70 in group C was higher than that in group B(P

ObjectiveTo investigate the mechanism of cerebral white matter damage in premature rats induced by intrauterine infection.Methods30 mature pregnant Wistar-Imamichi rats were divided into the experimental group (n=18) and control group (n=12). The rats of the experimental group were injected with lipopolysaccharide (LPS) 0.2 mg/kg in abdomen on 15th and 16th day after gestation. Those of the control group were injected with distilled water in equal volume. 45 premature rats born in the experimental group and 45 full-term rats born in the control group were tested with RIA for interleukin-6 (IL-6). The brain tissues of another 47 premature rats and 41 full-term rats were stained with HE method. Finally water content in brain tissue were tested in 50 premature rats and 50 full-term rats.ResultsIL-6 concentration of brain tissue of the premature rats in the experimental group was significantly higher than that in the control group ( P<0.01); and water content of the premature rats' brain was also higher than that of the full-term rats ( P<0.01). The edema of periventricular white matter, loose neuropil, decreased cell numbers, broaden intercellular space, and cell swelling and rupture were found in the brain tissue of the premature rats, no abnormal form and structure were found in the control group.ConclusionWhite matter damage of premature rats can be caused by endotoxin, and accompanied by IL-6 and water contents increasing.

Objective To evaluate abdominal imaging in AIDS.Methods The imaging examinations(including US,CT and MR)of 6 patients with AIDS associated abdominal foci were analysed retrospectively.All the cases were performed US,and CT scan,of which 4 performed enhanced CT scan and 1 with MR.Results Abdominal tuberculosis were found in 4 patients,including abdominal lymph nodes tuberculosis(3 cases)and pancreatic tuberculosis(1 case).The imaging of lymph nodes tuberculosis typically showed enlarged peripheral rim enhancement with central low-attenuation on contrast-enhanced CT. Pancreatic tuberculosis demonstrated low-attenuation area in pancreatic head and slightly peripheral enhancement.Disseminated Kaposi's sarcoma was seen in 1 case:CT and MRI scan demonstrated tumour infiltrated along hepatic portal vein and bronchovascular bundles.Pelvic tumor was observed in 1 case:CT scan showed large mass with thick and irregular wall and central low attenuation liquefacient necrotic area in the pelvic cavity.Conclusion The imaging findings of AIDS with abdominal foci is extraordinarily helpful to the diagnosis of such disease.Tissue biopsy is needed to confirm the diagnosis.

Objective To assess the indication and safety of surgical resection of the pregnancy by hysterotomy (SRPH) and hysterectomy for cesarean scar pregnancy (CSP). Methods A retrospective study of women with CSP was conducted at the Women′s Hospital, School of Medicine, Zhejiang University, from Jan. 2003 to Mar. 2016. The women underwent SRPH (SRPH group, n=35) and hysterectomy (Hysterectomy group, n=14) were included. The gestational age (GA), size of gestational mass(GM), level of serum β-hCG, previous treatments and clinical outcomes were analyzed. Results The median GA, the mean size of GM, median serum β-hCG level, median amount of blood loss, rate ot blood transfusion, rate of persistent CSP, and rate of motal status in SRPH group versus Hysterectomy group were 66 versus 84 days, (65 ± 22) versus (92±36) mm, 23755 versus 802 U/L, 400 versus 650 ml, 11%(4/35) versus 13/14, 49%(17/35) versus 12/14, 20% (7/35) versus 14/14, respectively (all P<0.05). In SRPH group, median amount of blood loss was 500 ml in patients with GA≥10 weeks versus 300 ml in patients with GA<10 weeks (P<0.05). Serious complication occurred in 7 patients: severe pelvic inflammation in 1 patient and hematomas in the uterine isthmus in 1 patient in SRPH group; severe pelvic inflammation in 2 patients and hemorrhagic shock and DIC in 3 patients in Hysterectomy group. No blaader damage occurred. Conclusions SRPH is effective and safe for patients with CSP with GA of 9-10 weeks, a diameter of 60-90 mm and stable hemodynamics. Hysterectomy is an alternative to SRPH for patiens in motal status with advanced GA more than 12 weeks.

Objective:To evaluate the expression of CXCR4 and the migration rate of bone marrow stromal CD34+cells in differ-ent risk groups with myelodysplastic syndromes (MDS) using correlation analysis. Methods: Forty MDS patients were divided into low-and high-risk groups based on the International Prognosis Scoring System (IPSS). The former was composed of 20 patients with IPSS<1.5, whereas the latter was composed of 20 patients with IPSS≥1.5. Bone marrow (BM) samples of these patients and 10 nor-mal controls were collected. CD34+cells were separated and purified. The expression of CXCR4 was determined by flow cytometry. The migration rate of CD34+cells on the chemotactic effect of SDF-1αand on the effect of bone marrow stromal cells were measured. Results:The expression rate of CXCR4 was higher in the high-risk MDS group than in the low-risk and control groups (P<0.000 1). No significant differences existed between the low-risk and the control groups (P>0.05). The migration rate of CD34+cells on the ef-fects of SDF-1αand marrow stromal cells were significantly increased in the high-risk MDS group compared with those in the low-risk and control groups (P<0.000 1). Migration rate of CD34+cells on the effect of marrow stromal cells was positively correlated with CX-CR4 expression (P=0.000 1). Conclusion:The CXCR4 expression and migration rates of CD34+cells on the effect of marrow stromal cells are significantly higher in the high-risk MDS group than in the low-risk group. Migration rate has a positive correlation with the CXCR4 expression, which further indicates that MDS is a heterogeneous group of hematopoietic stem cell malignancies. The expres-sion and function of SDF-1 and its receptor CXCR4 differ within each group with various risks. SDF-1 and CXCR4 may be involved in MDS pathogenesis.