To explore the plasticity of human amniotic mesenchymal cells(hAMCs) into cardiomyocyte-like cells,hAMCs were isolated from human amnion with collagenase digestion.Phenotype of the isolated cells was analyzed by flow cytometry(FCM).hAMCs were treated with 5-azacytidine and basic fibroblast growth factor(bFGF) to investigate their ability of differentiation into cardiomyocytes.The induced differentiated cells were evaluated by immunofluorescence for desmin and ?-actin expression and by RT-PCR for Nkx2.5,GATA-4 and alpha-myosin heavy chain(?-MHC) mRNA expression.The results showed that,after primary culture,hAMCs could reach a confluence of 80% with swirl like growth at 6 days.The cells proliferated rapidly after passages with a 100% confluence at 3~4 d.hAMCs were positive expression of CD44 and vimentin,but negative for CK19.After induced differentiation at 8~10d,the differentiated cells have close-up arranged with long spindle-shape.At 2 weeks and 4 weeks,induced cells expressed ?-actinin and cardiac-specific transcription factor Nkx2.5.Expressions of GATA-4 and desmin can be detected but ?-MHC can not in the hAMCs both before and after the induction.In conclusion,hAMCs have the ability of differentiation into cardiomyocyte-like cells,which means that hAMCs can be regarded as candidate cells for cellular cardiomyoplasty(CCM).

Computational epidemiology is a fast-developing and interdisciplinary research area .Through comprehensive computation-analysis of multi-uncertain factors affecting the epidemic process , this method may add to our knowledge about epidemic patterns and help design reasonable response plans and emergency strategies .This article briefly summarizes the idea and theory of computational epidemiology based on related researches in the recent years , introduces the application of this method in case of smallpox bioterrorism and influenza pandemic , and predicts the development of this area .

BACKGROUNDThe technique of intersphincteric resection of tumors combined with coloanal anastomosis has been used to avoid permanent colostomy for patients with a rectal cancer located < 5 cm from the anal verge. This study aimed at assessing the preservation of continence function of the residual rectum and the clinical prognosis of patients with lower rectal cancer after intersphincteric resection using a prolapsing technique.

METHODSThis study included patients with the following inclusion criteria: (1) pathological evidence of rectal cancer and the tumors within distal margins located 5 cm or less from the anus by preoperative endoscopic examination; (2) no evidence by MRI of infiltration of either the external sphincter, puborectalis or the levator muscle; (3) the patients are eligible for intersphincteric resection and lower coloanal anastomosis with a preoperative biopsy showing the tumors with well-to-moderate differentiation. From January 2000 to June 2004, 23 patients with low rectal cancer were included in this study. We used the standard abdominoperineal approach to perform radical resection of tumors with excision of the mesorectum and total or part of the internal sphincters. The patients were followed for assessment of the function of the residual rectum and of cancer recurrence after the operations.

RESULTSThe median tumor distance from the anal margin was 4.5 (range 3.5 - 5.0) cm and the mean distal surgical margin 1.6 (range 1.0 - 2.0) cm. Cancer was classified into Stage I (30.4%), Stage II (47.8%), and Stage III (21.7%) according to the TNM classification. Two patients developed anastomotic fistula after the surgical resection and 2 patients (8.7%) developed later stages of anastomotic stricture at the site of coloanal anastomosis. The median follow-up period was 31.5 months (range 12 - 54) and 2 patients (8.7%) developed local recurrence. Three deaths were associated with distal organ metastasis. Twenty patients (87.0%) have maintained competence to control solid or liquid stool and the capacity of flatus continence after the surgery. Among these patients, 2 patients were able to control solid stool and occasionally lose continence of liquid stool. And only 1 patient (4.4%) has retained partial rectum function with good continence of solid stool but not liquid after the operations. Average times of defecation per day of 3, 6, 12, 24 and 36 months after the surgery were 13.1, 4.7, 3.1, 2.9, and 3.2 times/day. Anal manometer measurements showed a decrease of pressure during the resting time after intersphincteric resection and this change remained during the period of follow-up. The maximum squeeze pressure was improved after an initial decrease after the surgery.

CONCLUSIONSMore residual rectum function after the surgery may be preserved by intersphincteric resection of low rectum cancer. At the same time this technique is safe with few postoperative complication and low tumor recurrence after the surgery.

Digestive System Surgical Procedures ; methods ; Female ; Follow-Up Studies ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Postoperative Complications ; etiology ; Prognosis ; Rectal Neoplasms ; mortality ; pathology ; physiopathology ; surgery ; Rectum ; pathology

OBJECTIVETo investigate the characteristics of transurethral partial cystectomy with a 2 µm laser in diagnosis and treatment for the bladder submucosal lesions in adults.

METHODSNine patients with suspected pathological diagnosed bladder submucosal lesions in out-patient department were diagnosed and treated transurethral with a 2 µm laser under sacral block between August 2009 and December 2010. The diameters of tumors were 1.5 - 2.5 cm. A 2 µm laser was used to incise the full-thickness bladder wall around the tumors. The entire bladder wall was peeled between the detrusor muscle layer and outer connective tissues. Tumors with bladder wall at the base were removed together and sent for pathological examination. The surgical procedures, intraoperative hemorrhage, intraoperative and postoperative complications were observed, pathological diagnosis and postoperative follow-up were performed.

RESULTSAll operations were successful. Mean operative time was 36.4 minutes (range 25 to 47 minutes), perioperative blood loss was minimal. There was no obturator nerve reflection and no hemorrhage detected after surgery. Postoperative pathological diagnosis included leiomyoma in 3 cases, pheochromocytoma in 3 cases, endometriosis in 1 case and metastatic bladder cancer in 2 cases.

CONCLUSIONSTransurethral partial cystectomy with a 2 µm laser can diagnose and treat bladder submucosal lesions. The procedures are effective and safe. Patients could get accurate pathological diagnosis without further painful and some bladder tumors can be treated by minimally invasive surgery.

Adult ; Aged ; Cystectomy ; methods ; Female ; Follow-Up Studies ; Humans ; Laser Therapy ; Male ; Middle Aged ; Treatment Outcome ; Urinary Bladder Neoplasms ; surgery

·AIM:To investigate and analyse the prevalence and risk factors associated with diabetic retinopathy severity in Qingdao. ·METHODS: This survey consisted of the 2 following parts: 2859 community residents aged >60 years old and 4275 patients with T2DM who were older than 30 years old in Qingdao. Ophthalmic examinations were performed on all patients. A questionnaire was used to obtain the patient's age and gender, the duration of diabetes mellitus(DM), glycaemic control and their knowledge of diabetic retinopathy ( DR ). Blood pressure and haemoglobin levels were recorded. All included patients underwent a comprehensive ophthalmic examination that included a fundus examination and retinal photographs and that assigned a grade for the severity of retinopathy according to the Early Treatment Diabetic Retinopathy Study (ETDRS) severity scale. Patients with severe non-proliferative or proliferative diabetic retinopathy and clinically significant macular edema ( CSME ) required ophthalmic therapy were assigned to the need-treatment group, while the remaining patients with DR were assigned to the need-observation group. Correlation and regression analyses were performed to determine the required-treatment rate and risk factors for DR. Logistic regression models were used to estimate odds ratios (OR) and 95% confidence intervals (CI) after adjustment for age, gender and the duration of diabetes. ·RESULTS: DR was present in 334 (11. 68% ) of the 2859 community residents aged > 60 years old and 1097 (25. 66% ) of the 4275 hospital patients with T2DM, and 48 (14. 81% ) of the residents and 172 ( 15. 68% ) of the hospital patients required ophthalmic therapy. In univariate and multivariate logistic analyses, factors including the age of the patients (51-60 years old: OR, 1. 68; 95% CI, 1. 21-1. 72; 61-70 years old: OR, 1. 55;95% CI, 1. 38-1. 76), the duration of diabetes (11-15 years:OR, 2. 61; 95% CI, 1. 51-4. 72; >15 years: OR, 4. 15; 95% CI, 2. 32-5. 77), glycaemic control (medium: OR, 2. 51;95%CI,1.98-3.92;poor:OR,4.69;95%CI,3.39-6.95), and knowledge of DR ( did not understand: OR, 1. 45;95%CI, 1. 21-1. 95) were significantly associated with the required-treatment rate in DR, while gender, low and advanced age ( 31-50 years old and >70 years old ), duration of disease (<10y), hypertension, and insulin treatment did not. ·CONCLUSION: The prevalence rate and the required-treatment rate in DR in Qingdao are relatively high. Being aged 51-70 years old and having a duration of diabetes>10y, poor glycaemic control and a lack of knowledge of DR were found to be potential risk factors that increased the rate of required ophthalmic therapy in patients with DR. In patients with T2DM who were aged 51-70 years old, we found that focusing on using science and education to strengthen the patients' knowledge of DR, establishing specifications for a community DR screening system, and effectively implementing early intervention in the community of DR - affected individuals were particularly important for preventing and controlling the high DR prevalence and the high rate of DR-associated blindness

BACKGROUNDHepatic alveolar echinococcosis (AE) is a parasitic disease in humans and caused by the Echinococcus multilocularis (Em). Orthotopic liver transplantation (OLT) may be the only effective treatment for end-stage hepatic AE. However, in some AE patients, extrahepatic Em can not be completely eliminated after OLT. We aimed to study whether the immunological changes caused by Em evasion may influence the rejective response.

METHODSRat modles of AE were established by injecting the Em suspension into abdomen of Brown Norway (BN) rats. Three months later, in the experimental group, the liver was transplanted from Lewis (LEW) rats to Em-infected BN rats. In the control group, transplantation was from LEW rats to healthy BN rats. Liver tissue and peripheral blood (PB) samples were collected on days 1, 3, 5, and 7 after OLT. Liver tissue was analyzed after hematoxylin and eosin (H&E) staining; numbers of CD4, CD8, and CD28 on peripheral blood cells were detected by flow cytometry; and expression of the chemokine fractalkine (Fkn) was detected by reverse transcription PCR (RT-PCR). Interleukin-10 (IL-10) was measured in the serum by enzyme-linked immunosorbent assay (ELISA). In every group, eight BN rats were retained for observing survival time.

RESULTSThe survival times of recipients in the experimental group were prolonged compared with those in the control group. The rejective response occurred later and was milder in the experimental group. percentage of CD4, CD8, CD28 T-cells and Fkn mRNA expression were lower in the experimental group. While the serum IL-10 levels were higher in the experimental group than those in the control group.

CONCLUSIONSAcute rejective response after OLT was attenuated in the rats with Em infection, and the recipients` survival time was prolonged. Em may play a role in this process by elevating IL-10 secretion, decreasing the effector T cells, inhibiting the expression of Fkn, which lead to reduce the inflammatory cells infiltration into the liver.

Animals ; CD28 Antigens ; metabolism ; CD4-Positive T-Lymphocytes ; metabolism ; CD8-Positive T-Lymphocytes ; metabolism ; Echinococcosis, Hepatic ; mortality ; surgery ; therapy ; Echinococcus multilocularis ; pathogenicity ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Graft Rejection ; immunology ; Interleukin-10 ; blood ; Liver Transplantation ; adverse effects ; Rats ; Reverse Transcriptase Polymerase Chain Reaction

The aim of this study was to evaluate whether human placenta CD133(+) cells have an ability to reconstitute long-term hematopoiesis. Magnetic-activated cell sorting (MACS) was applied to enrich human placental CD133(+) cells. The isolated human placental CD133(+)cells of four different densities were established by limiting-dilution assay and primary fetal bone marrow stromal cells separated from bone marrow as feeder layer cells were co-cultured in long-term culture system so as to observe the incidence of long-term culture initiating-cells (LTC-IC) and their ability of proliferation and differentiation.The results showed that human placenta derived CD133(+) cells contained LTC-IC with frequency of 1/645 which have an ability to proliferate and differentiate into granulocyte/macrophage colony-forming units (CFU-GM) and mixed colony-forming units (CFU-Mix). In all LTC-IC positive wells, 71.43% form only CFU-GM and 28.57% display both CFU-GM and CFU-Mix formation. It is concluded that human placental CD133(+) cells possess LTC-IC with colony-forming capacity of hematopoietic early progenitor cells.
AC133 Antigen ; Antigens, CD ; immunology ; Cell Culture Techniques ; methods ; Cell Differentiation ; Cell Separation ; Colony-Forming Units Assay ; Female ; Glycoproteins ; immunology ; Hematopoietic Stem Cells ; cytology ; Humans ; Peptides ; immunology ; Placenta ; cytology ; immunology ; Pregnancy

OBJECTIVETo study whether human placenta contains hematopoietic stem/progenitor cells (HSPCs), and analyze phenotypes of lymphocyte subpopulations in the placenta.

METHODSNucleated cells from fresh human placenta were analyzed for phenotypes of HSPCs and lymphocyte subpopulations by flow cytometry (FCM). And CD(34)(+) cells were sorted from human placenta nucleated cells by FCM or MiniMACS.

RESULTS(1) CD(34)(+) cells, CD(34)(+)/CD(38)(+) cells, and CD(34)(+)/CD(38)(-) cells from a human placenta were 8.8, 4.6 and 11.9 times higher than those from umbilical cord blood (UCB), respectively. (2) The yields and purity of CD(34)(+) cells isolated from human placenta by FCM sorting system were (63.05 +/- 10.14)% and (86.39 +/- 11.27)%, respectively. (3) Lymphocytes, T cells (CD(3)(+)/CD(2)(+)), B cells (CD(19)(+)), Th cells (CD(3)(+), CD(4)(+)), and Th/Ts ratio in the placenta tissue were apparently lower than those in the UCB, while the CD(8)(+)/CD(28)(-) T suppressor cells were higher in the placenta than in the UCB.

CONCLUSIONSHuman placenta is rich in HSPCs, and has important hematopoietic function in ontogeny. It is probable that human placenta would be graft resource for HSPCs transplantation. CD(8)(+)/CD(28)(-) T suppressor cells might play an important role in feto-maternal immunologic tolerance.

Cells, Cultured ; Female ; Hematopoietic Stem Cells ; cytology ; immunology ; Humans ; Lymphocyte Count ; Lymphocyte Subsets ; cytology ; immunology ; Male ; Placenta ; cytology ; immunology ; Pregnancy

The aim of this study was to establish the standard protocols for isolating and enriching hematopoietic stem/progenitor cells (HSPC) from human placenta tissue (PT). Single-cell suspension from of human PT was prepared by mechanical method combined with collagenase digestion. Mononucleated cells (MNC) derived from PT were separated by hydroxyethyl starch (6% HES), then the three cell subsets of different immunophenotypes (CD34(-), CD34(+)CD38(-), CD34(+)CD38(+)) contained in MNC were isolated by Magnetic Activated Cell Sorting (MACS). The cell immunophenotype of each sorting steps was analyzed by flow cytometer (FCM). The cell enrichment and recovery rate of each sorting step were calculated. The results showed that MNC could be harvested up to (12.30 +/- 3.51) x 10(8) from a single-cell suspension of human PT by mechanical method and collagenase digestion, no significant difference existed as compared with umbilical cord blood (UCB) initial sample [(8.86 +/- 5.38) x 10(8)], but the percentage of CD34(+) cells in MNC of human PT was (3.93 +/- 2.31)%, much higher than that in UCB [(0.44 +/- 0.29)%] (P < 0.001). recovery rate of MNC and CD34(+) cells from PT after separation with 6% HES were (45.3 +/- 11.7)% and (51.1 +/- 9.8)%, respectively. After MNC being sorted by MACS, the enrichment and recovery rate of CD34(+) cells in CD34(+) group were (73.4 +/- 14.1)% and (52.7 +/- 11.7)% respectively. It is concluded that the protocols established here for isolating and enriching hematopoietic stem/progenitor cells from human placenta can acquire HSPC with high abundance, enrichment and viability and may be a useful reference of isolating methods for future related study.
Antigens, CD34 ; analysis ; Cell Proliferation ; Cell Separation ; methods ; Hematopoietic Stem Cells ; cytology ; immunology ; Humans ; Immunophenotyping ; Placenta ; cytology

OBJECTIVETo study the expansion potential of megakaryocyte progenitor cells (MPC) from human placenta tissue CD133+ (PT-CD133+) cells.

METHODSPT-CD133+ cells were purified from mononuclear cells (MNC) by magnetic activated cell sorting (MACS) and seeded in serum-free liquid culture medium supplemented with thrombopoietin (TPO), interleukin-3 (IL-3), and stem cell factor (SCF) to expand MPC. At day 7, 10 and 14, the total cell number was counted and the dynamic changes of CD133, CD34, and CD41 antigens expression during ex-vivo expansion were analyzed by flow cytometry (FCM). PT-CD133+ cells at different expansion time were collected and cultured in collagen semisolid medium for colony forming units-megakaryocyte (CFU-MK) assay.

RESULTSPT-CD133+ cells could be optimally expanded at day 7 by 13 +/- 2 fold increase in serum-free liquid culture systems and the total cell number was expanded by 160 fold at day 14. With the expansion time going on, the expression of CD133, CD34 decreased and that of CD41 increased. The expanded megakaryocytes were immature and no sign of platelet formation. Both PT-CD133+ cells before and after expansion could form CFU-MK, the total number of CFU-MK peaked at day 10 of expansion by 54 +/- 10 fold increase.

CONCLUSIONHuman PT-CD133+ cells have a high capacity of MPC expansion, 10 days culture could give rise to the maximum number of CFU-MK.

AC133 Antigen ; Antigens, CD ; Cell Differentiation ; Cells, Cultured ; Female ; Glycoproteins ; Humans ; Megakaryocyte Progenitor Cells ; cytology ; Peptides ; Placenta ; cytology ; Pregnancy