Clinical evidence of hematopoietic restoration with umbilical cord blood (UCB) grafts indicates the UCB can be a useful source of hematopoietic stem cells for routine bone marrow reconstitution. Considering (10 +/- 5) x 10(8) nucleared cells per cord blood unit, there is a potential limitation for the use of cord blood in adults, which, however, can be overcome by ex vivo expansion of cells. A prerequisite for expansion is the significantly higher recovery of MNC, CD34+ cells and colony-forming cells (CFC) by thawing cryopreserved MNC. Cooling rate always acts as a critical factor that can affect the recovery of cells. Although the rate of - 1 degrees C/min is adopted in most of the cryopreservations, no data has been reported about the detailed effects of different cooling rates. The aim of the study was to reveal the different effects of cooling rates on cryopreservation of hematopoietic stem cells from cord blood. UCB samples were collected, and cryopreserved as mononuclear cells (MNC) with different cooling rates of - 0.5 degrees C/min, - 1 degrees C/min, - 5 degrees C/min, and the recovery and viability of MNC and CD34+ cells, the clonogenic capacity and the ex vivo expansion potential of UCB progenitor cells were evaluated after thawing. With - 1 degrees C/min cooling rate, the recovery of MNC reached 93.3% +/- 1.8% , viability 95.0% +/- 3.9% , recovery of CD34+ cells 80.0% +/- 17.9% , and clonogenic recovery were 87.1% +/- 5.5%, 88.5% +/- 8.9%, 86.2% +/- 7.4% for BFU-E CFU-GM CFU-MK, respectively. After 14 days of liquid culture, no significant difference was detected in CFC expansion between fresh and cryopreserved MNC cells with - 1 degrees C/min cooling rate, but this was not the case with - 0.5 degreesC/min and - 5 degrees C/min. In conclusion, it was demonstrated that controlling the rate at - 1 degrees C/min is more suitable for cryopreservation of hematopoietic stem cells than - 0.5 degrees C/min and - 5 degrees C/min.
Cell Survival ; physiology ; Cells, Cultured ; Cryopreservation ; methods ; Erythroid Precursor Cells ; cytology ; Fetal Blood ; cytology ; Flow Cytometry ; Granulocyte-Macrophage Progenitor Cells ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans

OBJECTIVETo detect the DNA damage of mice endometrial cells induced by carbon disulfide with single cell gel electrophoresis (SCGE) and explore the mechanism of embryo implantation disorder.

METHODSEndometrial cells, obtained by mechanical scrape, were used to test cell viability by trypan blue. Single cell suspension was exposed to the different concentrations of carbon disulfide (CS(2)) at four dose groups (0, 500, 1000, 2500 micromol/L). DNA damage was detected by SCGE. The SCGE results were analyzed by CASP software.

RESULTSDifferent dosages of CS(2) concentration induced different varying degrees of damage, forming typical normal cell and comet cell images. Compared with the control group, HDNA% decreased by 7.49%, 12.19% and 24.36% respectively, and TDNA%, TL, OTM increased by 7.13, 11.60, 23.18, 3.68, 5.98, 9.62, and 9.16, 16.84, 39.32 times respectively, in the groups of 500, 1000, 2500 micromol/L CS(2) (P < 0.01). Compared with the group of 500, 1000 micromol/L CS(2), TDNA%, TL, CL, TM, OTM in the group of 2500 micromol/L CS(2) increased by 1.98, 0.92, 1.27, 0.52, 0.37 and 0.17, 5.31, 1.90, 2.97, 1.26 times respectively(P < 0.01). Compared with the group of 500 micromol/L CS(2), TDNA%, TL, CL, TM, OTM in the group of 1000 micromol/L CS(2) increased by 0.55, 0.49, 0.16, 1.18, 0.76 times respectively (P < 0.01). The result of regression analysis showed that regression coefficients between HDNA%, TDNA%, TL, TM, OTM and the doses were -13.78, 13.78, 0.05, 4.38 and 3.23 respectively.

CONCLUSIONSCS(2) exposure could induce DNA damage in the endometrial cells of mice at the phase of implantation. The degree of DNA damage increases with the increasing CS(2) concentration. CS(2) might affect the implantation of embryo by doing harm to the endometrial cells.

Animals ; Carbon Disulfide ; toxicity ; Comet Assay ; DNA Damage ; Embryo Implantation ; drug effects ; Endometrium ; cytology ; drug effects ; Female ; Mice ; Mice, Inbred Strains

Objective To develop a method for the determination of total cholesterol in serum by isotope dilution liquid chromatography tandem mass spectrometry (ID-LC/MS/MS).Methods Serum samples were supplemented by addition of [3,4-~(13)C_2]-cholesterol,hydrolyzed with alcoholic sodium hydroxide and oxidized into cholest-4-ene-3,6-dione by chromic acid.The oxidation products were analyzed by LC/MS/MS using atmospheric pressure chemical ionization (APCI) source and detection modes of multiple reaction monitoring (MRM) and single ion recording (SIR).Signals (peak areas) of the internal standard were corrected for the contributions of cholesterol and the signal ratios of cholesterol to internal standard for the calibrations were linearly regressed against cholesterol concentrations.The resulted regression equation was used for the calculation of serum cholesterol concentrations.Results The correlation coefficients between the peak area ratios and cholesterol concentrations were 0.999 9 and higher.Under MRM mode,the average within-run CV of the results obtained on 3 serum samples was 0.95% (ranged from 0.92% to 0.99%) and the total CVwas 0.86% (0.82% to 0.89%),and under SIR mode,the within-run CV was 0.64% (from 0.54% to 0.77%) and the total CVwas O.69% (0.62% to 0.81%),respectively. Results on certified reference materials (SRM 1951 a Level Ⅰ and Level Ⅱ;GBW 09145 and GBW 09147) showed an average bias of 0.23% (0.14% to 1.00%) under MRM mode,and 0.24% (0.07% to 1.27%) under SIR mode.Conclusions An ID-LC/MS/MS method for serum cholesterol has been developed.It is specific and precise and may be used as a candidate reference method.

Objective To investigate the clinical efficacy of transcranial Doppler(TCD)-assisted interventional intra-arterial thrombosis in patients with acute ischemic stroke.Methods This clinical trial enrolled 22 patients with acute ischemic stroke(13 with occlusion of the middle cerebral artery within 6 h of symptom onset; 9 with occlusion of the base artery within 12 h of symptom onset); occlusive arterial cannula was performed and recombinant tissue plasminogen activator(rt-PA,20 mg)was injected to perform interventional intra-arterial thrombosis; and TCD ultrasonography of low frequency(2 MHz)and low intensity(0.25 W/cm2)was performed to assist the thrombosis.Cranial CT was performed again right after the operation and 24 h after the operation to observe the recanalization rate and intracranial hemorrhage; NIHSS scores and Barthel index scores were compared before and after the operation.Results The percentage ofrecanalization was 77.27％(17/22),including 22.73％(5/22)complete recanalization and 54.55％(12/22)partial recanalization; non-symptomatic intracerebral hemorrhage occurred in 3 patients and no symptomatic intracerebral hemorrhage was noted.NIHSS and Barthel index scores after the operation were obviously higher than those before the operation,indicating that favorable outcomes were achieved after thrombosis.Conclusion TCD-assited interventional intra-arterial thrombosis with rt-PA,significantly improving the recanalization rate of occlusive artery and remarkably reducing the happening ofintracerebral hemorrhage,can improve the prognosis and enjoy good clinical efficacy and safety in treatment of acute ischemic stroke.

OBJECTIVEThe main treatment method used for thoracolumbar fractures is open reduction and internal fixation. Commonly there are three surgical approaches: anterior, posterior and paraspinal. We attempt to compare the three approaches based on our clinical data analysis.

METHODSA group of 94 patients with Denis type A or B thoracolumbar burst fracture between March 2008 and September 2010 were recruited in this study. These patients were treated by anterior-, posterior- or paraspinal-approach reduction with or without decompression. The fracture was fixed with titanium mesh and Z-plate via anterior approach (24 patients), screw and rod system via posterior approach (38 patients) or paraspinal approach (32 patients). Clinical evaluations included operation duration, blood loss, incision length, preoperative and postoperative Oswestry disability index (ODI).

RESULTSThe average operation duration (94.1 min +/- 13.7 min), blood loss (86.7 ml +/-0.0 ml), length of incision (9.3 mm +/- 0.7 mm) and postoperative ODI (6 +/- 0.5) were significantly lower (P less than 0.05) in paraspinal approach group than in traditional posterior approach group (operation duration 94.1 min +/- 13.7 min, blood loss 143.3 ml +/-28.3 ml, length of incision 15.4 cm +/- 2.1 cm and ODI 12 +/- 0.7) and anterior approach group (operation duration 176.3 min +/- 20.7 min, blood loss 255.1 ml +/- 38.4 ml, length of incision 18.6 cm +/- 2.4 cm and ODI 13 +/- 2.4). There was not statistical difference in terms of Cobb angle on radiographs among the three approaches.

CONCLUSIONThe anterior approach surgery is convenient for resection of the vertebrae and reconstruction of vertebral height, but it is more complicated and traumatic. Hence it is mostly used for severe Denis type B fracture. The posterior approach is commonly applied to most thoracolumbar fractures and has fewer complications compared with the anterior approach, but it has some shortcomings as well. The paraspinal approach has great advantages compared with the other two approaches. It is in accordance with the concept of minimally invasive surgery and can replace most posterior approach operations.

Adolescent ; Adult ; Aged ; Female ; Fracture Fixation ; methods ; Humans ; Lumbar Vertebrae ; injuries ; Male ; Middle Aged ; Spinal Fractures ; surgery ; Thoracic Vertebrae ; injuries

BACKGROUNDRapid eye movement (REM) sleep behavior disorder (RBD) may be a risk factor for cognitive impairment in patients with Parkinson's disease (PD). However, little is known regarding the relation between the severity of RBD and the different domains of cognitive impairment. The aim of this study was: (1) to investigate the domains of cognitive impairment in patients with PD and RBD, and (2) to explore risk factors for PD-mild cognitive impairment (PD-MCI) and the relationship between RBD severity and impairment in different cognitive domains in PD.

METHODSThe participants were grouped as follows: PD without RBD (PD-RBD; n = 42), PD with RBD (PD + RBD; n = 32), idiopathic RBD (iRBD; n = 15), and healthy controls (HCs; n = 36). All participants completed a battery of neuropsychological assessment of attention and working memory, executive function, language, memory, and visuospatial function. The information of basic demographics, diseases and medication history, and motor and nonmotor manifestations was obtained and compared between PD-RBD and PD + RBD groups. Particular attention was paid to the severity of RBD assessed by the RBD Questionnaire-Hong Kong (RBDQ-HK) and the RBD Screening Questionnaire (RBDSQ), then we further examined associations between the severity of RBD symptoms and cognitive levels via correlation analysis.

RESULTSCompared to PD-RBD subjects, PD + RBD patients were more likely to have olfactory dysfunction and their Epworth Sleepiness Scale scores were higher (P < 0.05). During neuropsychological testing, PD + RBD patients performed worse than PD-RBD patients, including delayed memory function, especially. The MCI rates were 33%, 63%, 33%, and 8% for PD-RBD, PD + RBD, iRBD, and HC groups, respectively. RBD was an important factor for the PD-MCI variance (odds ratio = 5.204, P = 0.018). During correlation analysis, higher RBDSQ and RBDQ-HK scores were significantly associated with poorer performance on the Trail Making Test-B (errors) and Auditory Verbal Learning Test (delayed recall) and higher RBD-HK scores were also associated with Rey-Osterrieth complex figure (copy) results.

CONCLUSIONSWhen PD-RBD and PD + RBD patients have equivalent motor symptoms, PD + RBD patients still have more olfactory dysfunction and worse daytime somnolence. RBD is an important risk factor for MCI, including delayed memory. Deficits in executive function, verbal delayed memory, and visuospatial function were consistently associated with more severe RBD symptoms.

Aged ; Aged, 80 and over ; Cognitive Dysfunction ; etiology ; Humans ; Logistic Models ; Middle Aged ; Parkinson Disease ; complications ; REM Sleep Behavior Disorder ; complications

Objective To investigate and analyze the situation of urban and rural neglected children aged 3-6,in China,so as to provide basis for the analysis and comparison on relevant risk factors.Methods 1163 urban children aged 3-6 (with 49.6％ males and 4.5％ with minority ethnicity) were investigated from 25 cities of 14 provinces,autonomous regions and municipalities in the whole country.Multi-stage stratified cluster sampling method was used.Again,using the same sampling method,4096 rural children (of whom 50.6％ were males with 6.2％ as minorities) were chosen from 26 cities of 10 provinces or municipalities.Identification of children being neglected was based on “Child Neglect Evaluation Norms of Children Aged 3-6 Years in Urban/Rural China”.SPSS-Windows 13.0 was employed for data analysis.Scores,frequency/degrees,age,sex and types (physical,emotional,educational,safety,medical and social) of children under negligence on every group of the regions,were calculated.x2 test (Chi-Square) and Analysis of variance (ANOVA) were processed to determine the significance of their differences.Results The overall frequencies of negligence were 28.0％ and 53.7％ respectively among the urban and rural children aged 3-6,while the total degrees of negligence were 42.2 and 44.4 respectively.Significant difference was found between children from the urban and the rural areas (P＜0.05).Significant difference was also found between urban and rural children on every age group (P＜0.05).The frequencies of negligence among males were 32.6％ and 55.9％ respectively in urban and rural areas while among females,the figures appeared to be 23.7％ and 51.6％ respectively.The degrees of negligence were 42.7 and 44.6 among male while 41.8 and 44.3 among female children,in the urban or rural areas.Significant differences were found on male or female between urban and rural groups (P＜0.05).Frequencies of negligence in urban children aged 3-6 for the six types were from 5.1％ to 12.9％,with the frequency in rural areas as 13.1％-26.6％.Significant difference was found between urban and rural group for any other type (P＜0.05),in addition to the safety type.The degrees of negligence in urban children aged 3-6 for the different type were between 39.4 and 43.4,while in the rural areas as from 36.5 to 48.2,with significant difference for every type (P＜0.05).The degrees of negligence related to education,emotion,or physical strength were more serious on children from the urban than from the rural areas.The highest frequency of child negligence was seen in the single-parent families on both urban and rural groups (42.9％ and 60.0％ respectively),with no significant difference found (P＞0.05).The urban and rural children aged 3-6 were mainly involved in single item of negligence,with incidence rates as 16.5％ and 22.7％ and proportions as 58.9％ and 45.1％ respectively,despite the factors as age or sex.Conclusion There were large differences on the situation of negligence between the urban and rural children aged 3-6.The frequencies and degrees of negligence in every age group and different sex for children living in the rural areas were higher than those urban children.The frequency of negligence among boys was higher than girls for both urban and rural areas.The rural children had suffered more serious negligence than the urban children at any other type,in addition to the ‘ safety'.Both urban and rural children had the highest frequency of negligence in single-parent family,and were mainly suffered from single item of negligence.

OBJECTIVETo evaluate the effects of nasal mometasone furoate on moderate to severe allergic rhinitis.

METHODSPatients with moderate to severe allergic rhinitis were enrolled and received mometasone furoate nasal spray 200 microg once daily for four weeks. Four hundreds and sixty-three patients completed the study. We weekly interviewed the patients to evaluate the symptoms, and the affection of disease on night sleep and daily life. Mini Rhinoconjunctivitis Quality of Life Questionnaire (Mini-RQLQ) and Nocturnal Rhinoconjunctivitis Quality of Life Questionnaire (NRQLQ) were used to evaluate the quality of life.

RESULTSThe individual and total symptomatic scores of week 1, week 2 and week 4 decreased compared with baseline. The scores of Mini-RQLQ and NRQLQ of week 1, week 2 and week 4 decreased compared with baseline too. The scores of the next visit were significantly lower than those of the previous visit (P < 0.01).

CONCLUSIONSA four-week administration of mometasone furoate nasal spray can effectively treat allergic rhinitis.

Administration, Intranasal ; Adolescent ; Adult ; Aged ; Anti-Allergic Agents ; administration & dosage ; therapeutic use ; Child ; Female ; Humans ; Male ; Middle Aged ; Mometasone Furoate ; Pregnadienediols ; administration & dosage ; therapeutic use ; Quality of Life ; Rhinitis, Allergic, Perennial ; drug therapy ; Surveys and Questionnaires ; Young Adult

OBJECTIVETo explore the status of child neglect among urban areas children and adolescents aged 3 - 17 years in China.

METHODSUsing multi-stage stratified cluster sampling method, 8001 children and adolescents were recruited from 30 cities of 14 provinces in December 2008. The investigation was carried based on The 3 - 6 years old children Neglect Norms of city in China and The 6 - 17 years old children Neglect Norms of city in China. SPSS 13.0 was employed for analyzing neglect rate and degree for group of areas, age, sex and neglect type (including neglect of physical, emotional, educational, medical, safety and social).

RESULTSThe degrees of child neglect for urban children aged 3 - 6 (children aged 6 were not in school), 6 - 8 (children aged 6 were school children), 9 - 11, 12 - 14 and 15 - 17 were 42.2 ± 6.9, 42.8 ± 8.4, 42.1 ± 9.1, 46.4 ± 9.2 and 49.7 ± 8.4(P < 0.05), the neglect rates were 28.0% (326/1163), 28.8% (431/1496), 27.2% (543/1962), 22.4% (373/1664) and 32.8% (563/1716) (P < 0.05), respectively. There were differences in six neglect types for each group; the neglect degrees and rates were high in the group aged 12 - 14 and 15 - 17 (46.4 ± 9.2 and 49.1 ± 8.4, 22.4% (373/1664) and 32.8% (563/1716), respectively); for each neglect type, the neglect degree of children aged 15 - 17 were highest (52.8 ± 8.0, 47.3 ± 11.6, 49.5 ± 10.8, 42.4 ± 10.3, 52.2 ± 16.0, 56.0 ± 10.1). The degree and rate of neglect in West were highest, Middle was next, the East was lowest (the neglect degrees were 45.4 ± 9.1, 45.0 ± 9.3 and 44.0 ± 8.8, P < 0.05;the rates were 30.7% (1340/4361), 25.0% (756/3024) and 21.2% (131/616), P < 0.05). Degree and rate of child neglect for children without siblings were lower than children with siblings (the neglect degrees were 44.2 ± 9.0 and 47.6 ± 9.1, P < 0.01; the rates were 26.1% (1572/6017) and 33.0% (655/1984), P < 0.01). Neglect degree of school children aged 6 years old (43.1 ± 7.3) was higher than children not in school (40.7 ± 7.8) (P < 0.01), but neglect rate was lower (the rates were 35.4% (60/169) and 21.3% (84/394), P < 0.01).

CONCLUSIONNeglect degree and rate of urban children and adolescents aged 3 - 17 years in China were both high, and neglect deserved more attentions.

Adolescent ; Child ; Child Abuse ; prevention & control ; statistics & numerical data ; Child, Preschool ; China ; Cross-Sectional Studies ; Female ; Humans ; Male ; Urban Population

BACKGROUND:Our preliminary findings have shown that stem cells have a certain effect on early formation of renal fibrosis and delay the occurrence of renal interstitial fibrosis.OBJECTIVE:To investigate the effect of exogenous adipose-derived stem cell transplantation on the formation of renal interstitial fibrosis in rats,and to explore the dose-effect relationship.METHODS:Fifty Sprague-Dawley rats were randomly divided into five groups.Rats in sham operation group were injected with PBS;and those in the other four groups were ligated to establish the model of renal fibrosis.After successful modeling,the model group was injected with PBS,while low dose group,middle dose group and high dose group were injected with 1 ×107/L,2x107/L,3x107/L adipose-derived stem cell suspension (0.1 mL),respectively.Rats were killed 14 days after injection.Hematoxylin-eosin staining and Masson staining were used to observe the degree of renal tubular interstitial injury and the relative area of the renal interstitium.Expression of alpha smooth muscle actin and transforming growth factor beta1 was detected by immunohistochemistry.RESULTS AND CONCLUSION:(1) Hematoxylin eosin staining:The model group showed typical glomerular and tubulointerstitial changes;the low,middle and high dose groups had a certain degree of glomerular and tubulointerstitial changes,but the severity of injury in these three groups,especially in the high dose group,was significantly milder than that in the model group.(2) Masson staining:In the model group,the renal interstitium was widened and the collagen fibers were deposited;in the low,middle and high dose groups,the degree of renal interstitial expansion was lower than that in the model group,and moreover,the degree of renal interstitial expansion in the middle and high dose groups was lower than that in the low dose group (P ＜ 0.05).(3) Immunohistochemical staining:Compared with the sham operation group,the expression of alpha smooth muscle actin and transforming growth factor beta1 in the model group was higher than that in the sham operation group (P ＜ 0.05).Compared with the model group,significantly decreased expression of alphasmooth muscle actin was found in the high dose group (P ＜ 0.05) and significantly reduced transforming growth factor beta1 expression was observed in the high,middle,and low dose groups (P ＜ 0.05),especially in the middle and high dose groups.To conclude,exogenous adipose-derived stem cell transplantation can delay the formation of early renal fibrosis in a dose-effect manner,which may be realized by changing the signal pathway state induced by transforming growth factor beta1.