In order to set up a base for stem cells to be widely used in clinical medicine, we tried to optimize, in this study, the technique that induces human mesenchymal stem cells (hMSCs) to differentiate into neural stem cells by using cerebrospinal fluid (CSF) from the different groups. After the induction, presence of neural stem cells was confirmed with microscope observation, flow cytometry analysis, immunohistochemistry and fluorescent immunohistochemistry. At the same time, we also compared and analysed the data of the number of stem cells when it totally met the requirements for clinical treatment and the days required. At last, we confirmed that hMSCs could be induced to differentiate into neural stem cells, and that the number of cells totally met the requirements for clinical treatment. But there were some differences both in the number of cells and the days required. Among the groups, the group that marrow mesenchymal stem cells from patients own induced by CSF from healthy volunteers used the shortest time and the quantity of the cells was significantly higher than those of the others.
Cell Differentiation ; Cerebrospinal Fluid ; chemistry ; Culture Media ; chemistry ; Flow Cytometry ; Humans ; Immunohistochemistry ; Mesenchymal Stromal Cells ; cytology ; Neural Stem Cells ; cytology

Objective To establish a molecular technique of internal transcribed spacer (ITS) sequencing to identify pathogenic fungi species from the fungal sinusitis tissues. Methods Total 270 sinusitis tissues samples were collected by endoscopic surgery from 2006 to 2008. The histopathology, organize spring clip culturation and ITS region (ITS region region of fungal rRNA, including ITS1-5. 8S rRNA-ITS2) sequencing were employed simultaneously. And then to evaluate the ITS sequencing as the tool for identification of pathogenic fungi directly from clinical samples. Results Of the 270 samples, histopathology positive rate was 80.0% (216/270) , organize spring clip positive rate was 80.0% (216/ 270), fungal culturation positive rate was 53.0% (143/270) , ITS region sequencing positive rate was 63. 0% [ (134 +28 +8)/270], There were 22 species and 6 genera identified by fungal culturation, and 32 species identified by ITS region sequencing. Conclusion ITS region sequencing will become a applicable tool in clinical laboratory in future.

ObjectiveTo study the imaging characteristics of SPN of adenocarcinoma (ASPNs) on 18F-FDG PET/CT.MethodsThe morphological and metabolic features of 35 ASPNs on FDG PET/CT were retrospectively reviewed.SUVmax (SUV) was measured and ΔSUVmax was calculated according to ΔSUVmax =(SUVmax on delay imaging - SUVmax on early imaging)/SUVmax on early imaging × 100％.Statistical analysis was performed by software SPSS 11.5 using t-test,analysis of variance and Fisher exact test.Results( 1 ) Fifteen ASPNs (42.86％,15/35) presented as nodular pattern on FDG PET imaging,while 20 (57.14％,20/35) as lamellar,cloudy or ill-defined patterns.The SUVmax of these ASPNs followed a descending order of nodular,lamellar,cloudy and ill:defined on both early and delay imaging (F =30.696 and 24.758,both P＜0.001).(2)There were 54.29％ (19/35) ASPNs with SUVmax ≥2.5 and 45.71％ (16/35) ASPNs with SUVmax ＜2.5.(3) Of 35 ASPNs,24(68.57％ ) were solid nodules and 11(31.43％) were ground glass nodules with SUVmax =4.54 ±2.69 and 1.30±0.87,respectively (t =-5.234,P ＜O.001 ).(4) The SUVmax of ASPNs on delay FDG imaging (4.22 ±3.52) was significantly higher than that on early imaging (3.49 ±2.72) (t =-4.021,P ＜0.001 ).However,SUVmax was dependent on SUVmax on the early imaging:when SUVmax ≥2.5,ΔSUVmax was positive in 94.74％ (18/19) of ASPNs; while SUVmax ＜2.5,ΔSUVmax was positive in 56.25％ (9/16) of ASPNs (P =0.013).(5) Of 31 ASPNs with cell differentiation data,there were 10/17 well-differentiated ASPNs and 13/14 poorly-differentiated ASPNs with positive ΔSUVmax ( P =0.045 ).The average SUVmax of well-differentiated ASPNs was significantly lower than that of poorly-differentiated ASPNs ( 1.70 ± 1.51 vs 4.91 ± 2.69,t =- 3.951,P ＜ 0.001 ).Conclusions The morphological and metabolic features of ASPNs are diversified.It is common for ASPN to present with SUVmax ＜ 2.5.ΔSUVmax may be helpful for differentiating malignant from benign SPNs.

Objective To evaluate the lymphoscintigraphic imaging characteristics for the patients with lower limb lymphedema and to establish a novel grading system for the injury to lower limb lymphatic system. Methods One hundred and sixty six consecutive patients (332 lower limbs) with lower limb lymphedema after surgical and(or) radiotherapy treatment for gynecological cancer were recruited into this retrospective study. The lymphoscintigraphy studies were performed after subcutaneous injection of 111～185 MBq (0. 1～0. 15 ml) of 99Tcm-DX into the webbed space between the first and second toes of both feet. Based on the integrity of lymphatic vessel and the extension of dermal diffusion on lymphoscintigram,the lymphatic injury to the lower limb was graded as 0, 1,2 and 3 respectively. The lymphedema of the limb was staged as 0, Ⅰ , Ⅱ a, Ⅱ b, Ⅲ by the standard of Consensus Document of the International Society of Lymphology (ISL). Chi square test was carried out to validate the established grading system for the assessment of the injury to the lower limb lymphatic system. Results The lymphoscintigraphic imaging characteristics included lymphatic blockage, dermal backflow, no visualization of lymphatic or lymph node, lymphocele and lymph fistula in the lower limb, pelvis and abdomen. There were 65 (19.6%), 71 (21.4%),131 (39.5%), 62 (18.7%) and 3 (0.9%) limbs staged as 0, Ⅰ , Ⅱa, Ⅱb, and Ⅲ for lymphedema while 36(10.8%), 79(23.8%), 116(34.9%) and 101 (30.4%) limbs graded as 0, 1, 2, and 3 for lymphatic injury. There was a statistically significant correlation between the grading methods (χ2 =313.483, P <0.001). The patients who underwent radiotherapy had a higher incidence rate of grade 2 and 3 (70.5%, 158/224) than those who underwent surgery (53.6%, 59/108) (χ2 = 9.662, P = 0.022).The patients with erysipelas had a higher incidence rate of grade 3(73.1%, 38/52) than those without erysipelas (43.9%, 50/114) (χ2= 12.238, P<0.001). The incidence rate of grade 3 increased with the duration of lymphedema after treatment: 36.6% (34/93) for less than 1.5 years, 72.3% (34/47) for between 1.5 to 5 years, and 76.9% (20/26) for more than 5 years (χ2 = 23.123, P<0.001). The grade of lymphatic injury showed no significant difference among 3 types of gynecological cancers (χ2 = 4.000, P =0.676), or between the patients with and without chemotherapy (χ2 =0.411, P=0.938). Conclusions Lymphoscintigraphy is a reliable modality to diagnose lower limb lymphedema after treatment for gynecological cancer. The injury grading system could provide objective assessment of the lymphatic damage.

OBJECTIVETo evaluate the performance of a new highly efficient and environment-friendly tissue cell fixatives for preserving the morphologies and properties of pleural and peritoneal effusions.

METHODSFifty-six specimens of tissue cells from pleural and peritoneal effusions were preserved using the new preservative or 95% ethanol. HE staining and Western blotting were employed to detect the morphologies and protein expression levels of CK, CEA and P53 of the cells after fixation.

RESULTSThe new preservative well preserved the morphologies of the cells from the pleural and peritoneal effusions, and the nuclei and cytoplasm were intact with little debris. The conventional preservative (95% ethanol) caused noticeable structural damage of the tissue cells, especially the cytoplasm where obvious debris were seen after fixation. CK, CEA and P53 protein expression levels in the cells were 91%, 86% and 88% after fixation with the new preservative, significantly higher than those (46%, 38% and 31%, respectively) in cells fixed with 95% ethanol (P<0.05).

CONCLUSIONThe new preservative is efficient and environment-friendly for preserving the morphologies as well as the proteins of tissue cells from pleural and peritoneal effusions well, demonstrating its potential in tissue cell fixation and preservation.

Ascitic Fluid ; cytology ; Biocompatible Materials ; Cytoprotection ; Humans ; Materials Testing ; Tissue Fixation ; methods

OBJECTIVETo investigate the effects of intensive insulin therapy on inflammatory response and prognosis of patients with severe trauma.

METHODSEighty severely injured patients were divided into intensive insulin therapy group (n = 40, IT) and routine therapy group (n = 40, RT) in random pair. At the time of admission, a continuous infusion of insulin (2-4 U/h) was pumped into the patients of IT group to maintain blood glucose level at 6-8 mmol/L. Patients in RT group were given routine treatment without administration of insulin. Fever, organ injury, and mortality of patients in 2 groups were recorded. Venous blood was drawn from patients of 2 groups on the morning of post treatment day (PTD) 1, 3, 5, and 7. Values of TNF-alpha, C-reactive protein (CRP), IL-2, and IL-10 in plasma were assayed.

RESULTSHigh fever appeared in 9 patients in IT group, and WBC exceeded 10.0 x 10(9) for more than 3 days in 17 patients in this group, versus 20 and 29 patients respectively in RT group. Dysfunction of 1 organ appeared in 31 patients in IT group and 30 patients in RT group. Dysfunction of 3 organs appeared in 10 patients in IT group and 19 patients in RT group. Dysfunction of 4 organs appeared in 7 patients in IT group and 12 patients in RT group. In IT group, 4 patients died within 3 post-injury day (PID), and 1 patient died after PID 3 (total case fatality: 12.5%). In RT group, 5 patients died within 3 PID, and 4 patient died after PID 3 (total case fatality: 22.5%). Plasma levels of TNF-alpha and CRP of patients in IT group were significantly lower than those of patients in RT group on PID 3-7 ( P < 0.05 or P < 0.01), while levels of IL-2 and IL-10 of patients in IT group were significantly higher than those of patients in RT group (P < 0.05 or P < 0.01). Plasma levels of TNF-alpha (1.3 +/- 0.6 microg/L) and CRP (55 +/- 16 mg/L) of patients in IT group on PTD 7 were lowered to the trough level, and they were significantly lower than those of patients in RT group (3.0 +/- 0.8 microg/L, 89 +/- 20 mg/L, respectively, P < 0.01).

CONCLUSIONSIntensive insulin therapy can mitigate systemic inflammatory response and improve prognosis of patients with severe trauma.

Adult ; Aged ; Female ; Humans ; Inflammation ; Insulin ; administration & dosage ; Interleukin-2 ; blood ; Male ; Middle Aged ; Prognosis ; Wounds and Injuries ; therapy ; Young Adult

OBJECTIVETo analyze the status and trend of injury deaths in Chinese people, and provide basic evidence for injury interventions.

METHODSData came from 2004-2005 the 3rd national retrospective sampling survey of death cause and covered 31 province-level regions and 160 surveillance spots in the interior of China, Total 142 660 482 person years were investigated. To describe the status of injury deaths, the crude death rate, years of potential life lost (YPLL), working years of potential life lost (WYPLL) and the standardized death rate were calculated. The population used for standardization was from census in 2000 and each five-year was counted as an age group; To analyze the trend of injury deaths, the constitution of the death causes were calculated based on the data of 1991-2000 national disease surveillance system which covered more than 10 000 000 population and 145 surveillance spots.

RESULTSThe total number of residents in survey districts died of injury between 2004 and 2005 was 87 753 (male 59 664, female 28 089, urban 23 308, rural 64 445); the crude death rate of injury in China 2004-2005 was 61.51/100 000 and accounting for 10.10% of all deaths; the standardized death rate was 58.45/100 000, ranking the fourth among the main cause of death for Chinese people. The YPLL of injury was 1579.61 person years per 100 000 and the WYPLL was 1721.41 person years per 100 000. The crude death rate of injury was 81.76/100 000 in male and 40.31/100 000 in female; the standardized death rates were 79.96/100 000 and 36.25/100 000, respectively. Injury mortality in male was two times higher than that in female. The crude death rates of injury were 48.66/100 000 in urban area and 68.01/100 000 in rural area; the standardized death rate were 44.08/100 000 and 66.25/100 000, respectively; the mortality in rural area was 1.4 times higher than that in urban area. The mortality for the aged 15 - 44 was 48.94/100 000(35 497/72 531 671) and accounting for 40% of all deaths, injury was the first cause of death for the aged 15 - 44. During 2004-2005, the top five causes of death related to injury were traffic accidents, suicide, falls, drowning and poisoning; the cases were 29 669, 18 678, 10 901, 7752, 4857 respectively in survey districts; the crude death rate were 20.80/100 000, 13.09/100 000, 7.64/100 000, 5.43/100 000, 3.40/100 000 respectively. From 1991 to 2005, the proportion of all injury deaths due to traffic accident increased from 15.00% (1551/10 338) to 33.79% (14 792/43 774) which showed a rising trend, the proportion of all injury deaths due to suicide decreased from 26.66% (2756/10 338) to 20.46% (8599/43 774) and the proportion of all injury deaths due to fall increased from 5.15% (532/10 338) to 12.87% (5630/43 774).

CONCLUSIONInjury is the primary cause of death resulting in premature death among Chinese people, traffic accident is the first cause of injury death. Since 1990s, the pattern of injury mortality of Chinese people has changed.

Accidents ; mortality ; statistics & numerical data ; Adolescent ; Adult ; Aged ; Cause of Death ; trends ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Young Adult

OBJECTIVETo identify the localization of hair follicles stem cell (HFSC) in different stages of hair and explore the differentiating capacity of HFSC into epidermis in vitro.

METHODSHFSC were detected by K19 immunostaining in normal human skin. Then, the isolated HFSC through enzyme digestion were seeded on dermal equivalent (DE) and cultured between the air-liquid interfaces for 14 days. Skin-equivalents was harvested and used for evaluation.

RESULTSHFSC mainly located in outer root sheet in hair follicle and human anagen hair follicles containing two distinct reservoirs for K19-positive cells located in the bulge and bulb of the follicle. These two reservoirs fused in line of outer root sheets during the catagen-telogen transition phase and individualized again in the newly forming anagen hair follicle. Based on DE, growing HFSC built a multilayered and confined epidermis.

CONCLUSIONHFSC located in outer root sheets can promote hair cycle and differentiate into epidermis in vitro.

Cell Differentiation ; physiology ; Cells, Cultured ; Epidermis ; cytology ; Hair Follicle ; cytology ; Humans ; Stem Cells ; cytology

PURPOSE: To investigate the efficacy and safety of umbilical cord blood (UCB) infusion (UCBI) plus immunosuppressive therapy (IST) treatment in comparison to IST treatment, as well as predictive factors for clinical responses, in severe aplastic anemia (SAA) patients. MATERIALS AND METHODS: Totally, 93 patients with SAA were enrolled in this cohort study. In the IST group, rabbit antithymocyte globulin (r-ATG) combined with cyclosporine A (CsA) was administered, while in the IST+UBCI group, r-ATG, CsA, and UCB were used. RESULTS: After 6 months of treatment, UCBI+IST achieved a higher complete response (CR) rate (p=0.002) and an elevated overall response rate (ORR) (p=0.004), compared to IST. Regarding hematopoietic recovery at month 6, platelet responses in the UCBI+IST group were better than those in the IST group (p=0.002), and UCBI+IST treatment facilitated increasing trends in absolute neutrophil count (ANC) response (p=0.056). Kaplan-Meier curves illuminated UCBI+IST achieved faster ANC response (p < 0.001) and platelet response (p < 0.001), compared with IST therapy. There was no difference in overall survival (OS) between the two groups (p=0.620). Furthermore, logistic regression analysis demonstrated that UCBI+IST was an independent predicting factor for both CR (p=0.001) and ORR (p < 0.001), compared to IST; meanwhile, very severe aplastic anemia (VSAA) and ANC could predict clinical responses as well. However, Cox proportional hazard regression indicated that VSAA (p=0.003), but not UCBI+IST, affected OS. Safety profiles showed that UCBI+IST therapy did not elevate adverse events, compared with IST treatment. CONCLUSION: UCBI+IST achieved better clinical responses and hematopoietic recovery than IST, and was well tolerated in SAA patients.
Anemia, Aplastic* ; Antilymphocyte Serum ; Blood Platelets ; Cohort Studies ; Cyclosporine ; Fetal Blood* ; Humans ; Logistic Models ; Neutrophils ; Umbilical Cord*

methods had some limitations. There was big difference between the results when using SEG and GGB, suggesting that we should try to combine GGB and SEG methods to get the better results.