Adult human bone marrow-derived mesenchymal stem cells (MSCs) were cultured on microcarriers in spinner flasks and were compared with those in conventional culture in 12-well plates. For the production of adherently growing MSCs, macroporous CultiSpher G gelatin microcarriers were used in concentration of 1 g/L. The cells were seeded in a density of 5 x 10(4) cells/ml in both spinner culture and conventional stationary culture. The result showed that after 7 days of cultivation a maximum viable cell concentration of 5.15 x 10(5) cells/ml was obtained in spinner culture. Whereas the cell density increased to a maximum of 1.675 x 10(5) cells/ml on day 5 in conventional stationary culture. Lactate was produced up to 12.06 mmol/L in spinner culture and up to 13.10 mmol/L in stationary culture, and glucose was consumed up to 7.38 mmol/L and 5.37 mmol/L respectively. The average lactate yield on glucose consumption in spinner culture was only 1.63, lower than that in stationary culture 2.44. This indicated that the energy metabolism in spinner culture was significantly more efficient than that in conventional culture. After spinner culture for 12 days, the MSCs maintain the characteristics of stem cells. It is concluded that the microcarrier culture system is a suitable way to expand the seeding cells for tissue engineering.
Adult ; Antigens, CD ; analysis ; Bone Marrow Cells ; cytology ; metabolism ; ultrastructure ; Cell Count ; Cell Culture Techniques ; instrumentation ; methods ; Cell Division ; Cell Survival ; Flow Cytometry ; Glucose ; metabolism ; HLA-DR Antigens ; analysis ; Humans ; Lactates ; metabolism ; Mesoderm ; cytology ; metabolism ; ultrastructure ; Microscopy, Electron ; Microscopy, Electron, Scanning ; Stem Cells ; cytology ; metabolism ; ultrastructure ; Time Factors

OBJECTIVETo investigate a new surgical method to treat unilateral limb lymphedem after radical mastectomy.

METHODS10 cases of upper limb lymphedema after radical mastectomy were treated using flap transfer (the lateral thoracic skin flap or latissimus dorsi musculocutaneous flap combined with liposuction).

RESULTSAfter the treatment, the upper limb perimeter reduced in varied degrees. Nuclear lymphatic radiography showed notable changes in lymphatic circulation. The effective results were steady during the follow-up of 3-18 months.

CONCLUSIONFlap transplantation combined with liposuction is a useful treatment for limb lymphedema from radical mastectomy.

Breast Neoplasms ; surgery ; Female ; Humans ; Lipectomy ; Lymphedema ; etiology ; surgery ; Mastectomy, Radical ; adverse effects ; Postoperative Complications ; surgery ; Surgical Flaps

OBJECTIVE: To establish a multiplex STR genotyping method for autosomal STR and Y-STR loci in forensic biological practice. METHODS: Widely used autosomal STR loci and Y-STR loci were selected. A set of PCR primers was designed, and a 5-dye fluorescent labeled STR multiplex PCR reagent kit was developed. RESULTS: A kit was developed which can simultaneously detect 15 autosomal STR loci, 10 Y-STR loci, and an Amelogenin. CONCLUSION The 15 autosomal STR plus 10 Y-STR kit in combination with capillary electrophoresis method was used to STR genotyping with accurate and reliable results. The new one-step testing kit can potentially be widely used in forensic cases and DNA databank in the future.
Amelogenin ; Chromosomes, Human, Y/genetics* ; DNA Primers ; Databases, Nucleic Acid ; Forensic Genetics/methods* ; Genotype ; Genotyping Techniques/instrumentation* ; Humans ; Indicators and Reagents ; Microsatellite Repeats ; Multiplex Polymerase Chain Reaction

Objective To investigate the prevention and rescuing measures of postoperative fatal bleeding induced by carotid blowout in head and neck tumors. Methods Seven cases with postoperative carotid bleeding treated from October 2003 to August 2009 were reviewed retrospectively. Of the patients, 6 were with common carotid blowout and one with internal carotid artery blowout. All patients underwent preor post-operative radiotherapy for primary head and neck tumours and 3 patients had neck defect repair with deltopectoral skin flap, frontal flap or free radial arm flap respectively. After carotid blowout bleeding, the patients were treated in time with X ray transcatheter intervention including transcatheter arterial embolization (TAE) and self-expanding covered stent implantation, followed by repairing the carotid region with appropriate myocutenous flaps. Results Of 7 patients with carotid blowout, 5 patients were successfully rescured with X ray transcatheter intervention, of them 2 with self-expanding covered stent implantation and 2 with TAE respectively, and other 2 patients died due to rapid bleeding. Of the successfully rescued patients, 2 patients were with the repair of carotid area by pectoralis major myocutaneous flap, one by submental flap and one by local flap, but another one not with flap repair. Follow-up showed the 3 patients rescued with self-expanding covered stent implantation were survival for 6,12,and 20 months, respectively, and the 2 patients rescued with TAE died of repeated carotid blowout in 2 and 13 months later, respectively. Conclusions The planned and timely X ray transcatheter intervention is an effective method to treat carotid blowout bleeding in the patients underwent head and neck tumour surgeries. Compared with TAE, selfexpanding covered stent implantation may be more reliable for restoring the blood supply of head and neck region, with less complications. One-stage repair of caratid region with myocutenous flap is of great importance to protect the carotid and to promote the wound healing.

OBJECTIVETo investigate the protective effect of Heme oxygenase-1 (HO-1) gene transfer on rat renal autograft against ischemia/reperfusion injury.

METHODSHO-1 recombinant adenovirus vectors were constructed and transduced into rat renal autograft by renal arterial perfusion. The renal autografts were transplanted orthotopically after store at 4 degrees C for 24 h, followed by contralateral native nephrectomy 5 d after transplantation. There were 25 rats in the control group. 5 h and 3 d after transplantation, reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemistry were used to detect the expression of HO-1 gene; enzyme-labeled immunosorbent (ELISA) was used to measure HO-1 protein content in the homogenate of renal autograft.

RESULTSThe intensity of HO-1mRNA expression at 3 h and 3 d after transplantation were 0.65 +/- 0.11, 0.86 +/- 0.17 in the experimental group and 0.09 +/- 0.01, 0.15 +/- 0.02 in the control group respectively. The differences between the two groups were significant (t = 14.38, 11.73, P < 0.05). HO-1 protein content at 3 h and 3 d after transplantation were significantly increased in the experimental group, as compared with the control group [(297 +/- 61) ng/g and (468 +/- 51) ng/g versus (98 +/- 30) ng/g and (155 +/- 31) ng/g; t = 8.27, 14.83, P < 0.05]. HO-1 transduced autografts had less renal ischemic injury and lower serum creatinine level compared with control animals (P < 0.05).

CONCLUSIONAdenoviral vector can successfully transduce rat kidneys with the HO-1cDNA, which can protect rat renal autografts from ischemia/reperfusion injury.

Adenoviridae ; genetics ; Animals ; Female ; Genetic Vectors ; Heme Oxygenase-1 ; biosynthesis ; genetics ; Kidney ; blood supply ; metabolism ; Kidney Transplantation ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; prevention & control ; Transfection ; Transplantation, Autologous

OBJECTIVETo study the protective effect of intensive insulin treatment on the myocardium of severely scalded rats, and to primarily explore its mechanism.

METHODSEighteen SD rats were divided into three groups, with 6 rats in each group. The rats in burn and intensive insulin group were inflicted with 30% TBSA full-thickness injury on the back. Isotonic saline containing 0.12 U/ml insulin solution, and 100 g/L glucose solution were infused into the rats in the intensive insulin group to keep plasma glucose at the level of 4.0 - 6.6 mmol/L (the total fluid amount was 2 ml x kg(-1) x 8h(-1)). In sham burn group,fluid was given according to physiological demand. The same amount of isotonic saline was infused into the rats in burn group. The venous blood was obtained for the detection of plasma glucose contents, and the left ventricular systolic pressure (LVSP) and left ventricular end-diastolic pressure (LVEDP) were recorded via aortic ventricle cannula before scald and at 1, 2, 3, 4, 5, 6 post-scald hours (PSH). The tissue of the left ventricle was harvested at 6 PSH for the detection of troponin T expression in myocardiocytes.

RESULTSPlasma glucose level was increased to (7.6 +/- 1.7) mmol/L - (8.4 +/- 4.7) mmol/L in burn group during 1-6 PSH, which was significantly higher than that in intensive insulin group (4.5 +/- 0.9) mmol/L - (5.2 +/- 1.3) mmol/L, P < 0.01). Compared with the intensive insulin group, LVSP was markedly decreased in the burn group (60 +/- 11 mm Hg vs 72 +/- 8 mm Hg, P < 0.05) at 1 PSH,whereas LVEDP was increased significantly (21.3 +/- 11.3 mmHg vs 11.7 +/- 5.2 mmHg, P < 0.05). Intensive insulin treatment could significantly inhibit the loss of troponin T protein in myofilaments of myocardium.

CONCLUSIONIntensive insulin treatment possesses a protective effect on myocardia function after severe burns, and it may be related to its preventive effect on the loss of contractile protein in cardiocytes.

Animals ; Blood Glucose ; metabolism ; Burns ; drug therapy ; metabolism ; Insulin ; administration & dosage ; therapeutic use ; Male ; Myocardial Contraction ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley ; Troponin T ; metabolism

OBJECTIVETo investigate of severe acute respiratory syndrome (SARS) convalescent stool shedding by RT-PCR.

METHODSOne hundred and three stool samples from 46 SARS patients were collected on May 16th, 20th, and 23rd, 2003. For each sample, RNA was extracted using commercial kit and 7 Nest RT-PCR using a 14-pair different SARS-associated coronavirus (SARS-CoV) special primers were carried out simultaneously.

RESULTSAmong these 46 SARS patients, 17 cases (37.0%) were stool SARS-CoV RT-PCR negative, and 29 cases (63.0%) were SARS-CoV RT-PCR positive. The duration of positive cases lasted (31.76 +/- 10.78) d (12-64 d). The longest stool shedding case in this study lasted 64 days. Two serial stool samples and for each sample 2 RT-PCR tests using different primers were positive in this case.

CONCLUSIONSOur study observed longest stool shedding of SARS patients to be 64 days after initial onset of SARS. The average stool shedding was 32 days. Hence it is important to think highly of SARS convalescent patient stool sterilization.

Adolescent ; Adult ; Feces ; virology ; Female ; Humans ; Male ; Middle Aged ; RNA, Viral ; analysis ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; SARS Virus ; isolation & purification ; Severe Acute Respiratory Syndrome ; transmission ; virology ; Time Factors

Objective To investigate the correlation between Chlamydia pneumonia (Cpn)infection and chronic obstructive pulmonary disease (COPD).Methods 82 patients with acute exacerbation COPD (AE-COPD) or stabilized COPD patients at outpatient visits,in the People' s Hospital of Jiangyin city from Aug.2010 to May 2012,together with 46 cases having stationary phase COPD and 38 healthy volunteers as control group,were involved in this study.Patients were bled 2 ml,on the next day of hospitalization while patients at emergency room were bled 2 ml immediately,but bled again on the 15th day.Serum was separated through cryopreservation and the Cpn antibodies (IgG,IgM and IgA antibodies) were detected,under micro-immunofluorescence.Results In terms of IgG in the three groups,the positive rates did not show significant differences (P＞0.05) but the GMT of the IgG in the AE-COPD group was significantly higher (P＜0.01) than that in the control group.IgA positive rate among the three groups; AE-COPD appeared the highest.There was no significant difference between the AE-COPD group and stationary phase COPD group (P＞0.05),however,there were significant differences between the AE-COPD group,the stationary phase COPD group and the control group (P＜0.01).In terms of GMT of IgA in the three groups,there was significant difference between the AE-COPD group and stationary phase COPD group (P＞0.05),but with significant difference between the AE-COPD group and the control group (P＞0.01).There was significant difference between stationary phase COPD group and the control group (P＞0.05).When comparing both the rates of acute infection and chronic infection on the AE-COPD groups with the control group,there appeared significant differences (P＜0.05,P＜0.01).When comparing the acute and chronic infection between the stationary phase COPD group and the control group,the rate of acute infection did not show significant difference (P＞0.05) while the chronic infection rate appear to have had significant difference (P＜0.01).Conclusion Cpn infection seemed to be closely related to the development of COPD.

OBJECTIVETo use a mathematical model and computer simulation to study transmission dynamics and control of schistosomiasis in mountainous regions of Sichuan.

METHODSBased on studies of schistosomiasis japonica transmission in 20 villages in mountainous regions of Sichuan, a mathematical model was developed to characterize the impact of local environmental factors on transmission intensity. The model integrated site-specific factors and was calibrated to field epidemiological data from 3 subset villages. The dichotomic method was then used to predict different control measures.

RESULTSThe study showed high variations in prevalence of infection and infection intensity across villages, ranging between 3%-73%, 0.1-100 epg (eggs per gram stool), respectively. Important factors including occupation of local residents, exposure to contaminated water, microclimatic characteristics were integrated in the model. The predictions of dichotomic models showed that continuing chemotherapy (coverage between 50%-60%) could reduce infection intensities to 30%-80%, but could not change local transmission potential; therefore, the termination of chemotherapy would be followed by bouncing back of transmission. Sustaining targeted environmental interventions through snail and parasite oval control at certain coverage (30%-50%, respectively) could reduce the transmission to relatively stable levels. The model predictions showed that an integrated control (e.g., including both chemotherapy and environmental interventions) could suppress the transmission to an undetectable level even interruption of transmission between 5-10 years.

CONCLUSIONThe study demonstrated the feasibility of using a dynamic model, calibrated to local data, to gain insights into complicated processes underlying the transmission and informing site-specific control strategies.

Adolescent ; Adult ; Animals ; Child ; Child, Preschool ; China ; Environment ; Humans ; Middle Aged ; Models, Theoretical ; Schistosoma japonicum ; Schistosomiasis japonica ; epidemiology ; prevention & control ; transmission ; Snails ; parasitology

OBJECTIVETo investigate the effects of leucine-rich repeats and immunoglobulin-like domains 3 (LRIG3) on the biological activity of glioblastoma cell line GL15.

METHODSGlioblastoma GL15 cells were cultured and transfected with LRIG3-EGFP plasmid. The location of LRIG3 in GL15 cells was observed with confocal microscopy. The proliferation and invasiveness of GL15 cells were detected with methyl thiazolyl tetrazolium (MTT) and Transwell methods respectively; the expression of epidermal growth factor receptor (EGFR) and LRIG3 mRNA and protein were detected with reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot respectively.

RESULTAfter transfection with the plasmid LRIG-EGFP, LRIG3 fusion protein was found in cytoplasm of GL15 cells and cell proliferative and invasiveness were reduced. The expression of EGFR and LRIG3 varied with the duration of EGF treatment (100 ng/ml): the expression of EGFR decreased while the expression of LRIG3 increased as time prolonged.

CONCLUSIONLRIG3 can inhibit the proliferation and invasiveness of glioblastoma cells and may be used as a target gene in gene therapy of glioblastoma.

Brain Neoplasms ; pathology ; Cell Proliferation ; Epidermal Growth Factor ; genetics ; Glioblastoma ; pathology ; Humans ; Membrane Proteins ; genetics ; metabolism ; Neoplasm Invasiveness ; Plasmids ; genetics ; RNA, Messenger ; genetics ; metabolism ; Receptor, Epidermal Growth Factor ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection ; Tumor Cells, Cultured