OBJECTIVETo investigate the prevalence of JAK2V617F and MPLW515L/K mutation in patients with slightly elevated platelets (BPC) or hemoglobin (Hb) not meeting the criteria of polycythemia vera (PV) or essential thrombocythemia (ET).

METHODSGenomic DNA from bone marrow or blood mononuclear cells was screened with allele specific polymerase chain reaction (AS-PCR) for JAK2V617F and MPLW515L/K mutation. The history of thrombosis was assessed retrospectively by patients files.

RESULTSOf 30 patients, 14 (46.7%) were positive for the JAK2V617F mutation, none of them had the MPLW515L/ K. Five of these 14 patients had a history of thrombosis. Follow-up results were available in 22 patients. Among them, 12 patients with JAK2V617F mutation turned out to be MPD in 6-24 months; only 2 out of 10 patients without this mutation evolved to MPD.

CONCLUSIONJAK2V617F mutation could be one of the diagnosis criteria of early MPD. No MPLW515L/K expression was found in early MPD.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Early Diagnosis ; Female ; Follow-Up Studies ; Humans ; Janus Kinase 2 ; genetics ; metabolism ; Male ; Middle Aged ; Mutation ; Myeloproliferative Disorders ; diagnosis ; genetics ; metabolism ; Receptors, Thrombopoietin ; genetics ; metabolism ; Young Adult

OBJECTIVETo investigate the friction and wear behavior of different human enamels.

METHODS24 enamel samples selected from aged, young permanent and faded deciduous teeth were classified into 3 groups and slid against artificial porcelain teeth in the presence of artificial saliva on an oscillating friction and wear test rig. The wear volume loss, microhardness and toughness of each group of the enamel specimens were measured, the wear scars were observed with a scanning electron microscope, and the elemental compositions of Ca, P, and Si of the wear scar and wear debris were determined with an energy dispersion spectrometer.

RESULTSThe wear volume losses of aged, young permanent and deciduous tooth enamels are (2.40 +/- 1.10) x 10(-12) m(3), (3.50 +/- 1.83) x 10(-12) m(3) and (4.86 +/- 2.49) x 10(-12) m(3). The data of aged tooth enamels are statistically greater than that of deciduous tooth enamels (P < 0.05). There is no significant difference between the wear volume loss of aged and young permanent tooth enamels or between the young permanent and deciduous tooth enamels (P > 0.05). However, the friction and wear behavior of each group of enamel specimens is different from each other.

CONCLUSIONSIn the present testing condition, the wear scars of each kind of enamel specimens is characterized by ploughing and cracking. The different wear resistance of the three kinds of enamels is attributed to the different microstructure of the enamel, while the hardness and toughness of the enamels are not correlated with the wear resistance.

Adolescent ; Age Factors ; Aged, 80 and over ; Child ; Child, Preschool ; Dental Enamel ; pathology ; Dentition, Permanent ; Female ; Humans ; In Vitro Techniques ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Tooth Abrasion ; pathology ; Tooth, Deciduous

Background: Preterm birth is a common cause of death in newborns and may result from many determinants, but evidence for the socioeconomic and environmental determinants of preterm birth in Tibetan women of childbearing age is limited. The aim of this study was to understand the current status of preterm birth in native Tibetan women and investigate the socioeconomic and environmental determinants. Methods: Data were drawn from a cohort study which was conducted from August 2006 to August 2012 in rural Lhasa, Tibet, China. A total of 1419 Tibetan pregnant women were followed from 20 weeks' gestation until delivery; the loss to follow?up rate was 4.69%. The incidence of preterm birth was estimated to show the status of preterm births in Tibet. Logistic regression models for longitudinal data were established, and odds ratios (ORs) together with 95% confidence intervals (CIs) were used to evaluate the association between the occurrence of preterm birth and 16 selected potential determinants based on the hierarchical conceptual frame. Results: The incidence of preterm birth was 4.58% (95% CI = 3.55–5.80%). After adjusting for health?related variables of the mothers and newborns, socioeconomic and environmental determinants associated with preterm birth included season (spring: OR = 0.28, 95%CI = 0.09–0.84; autumn: OR = 0.21, 95% CI = 0.06–0.69; and winter: OR = 0.31, 95% CI = 0.12–0.82) and calendar year of delivery (2010: OR = 5.03, 95% CI = 1.24–20.35; 2009: OR = 6.62, 95% CI = 1.75–25.10; and 2007–2008: OR = 5.93, 95% CI = 1.47–23.90). Conclusions: The incidence of preterm birth among native Tibetan women was low and there was a decreasing trend in recent years;however, it is still essential to strengthen seasonal maternal care, extend the spacing between pregnancies, and reinforce adequate maternal nutrition.

Aim To observe the effect of epidurally application of osthole on the model of nucleus pulposusinduced inflammatory radicular pain and the expression of p38 MAPK signaling related pathway in the spinal dorsal horn of rats.Methods The model of radicular pain was generated by putting nucleus pulposus to the L5 dorsal root ganglion (DRG).50％ MWT was measured using Von Frey filaments to calculate mechanical pain threshold before and after operation.50 μL of 20 g · L-1 osthole was administered epidurally in group Ost and 50 μL of 100 mL · L-1 DMSO in group DMSO at postoperative day (POD).The expression of phosphorylated p38 (p-p38),IL-18 and IL-18R in the lumbar spinal dorsal horn was detected by Western blot.IL-18 mRNA was assessed by real-time PCR.Results The mechanical pain threshold significantly decreased after operation (P ＜ 0.05),while the expression of protein p-p38 MAPK,IL-18,IL-18R and IL-18 mRNA was significantly different.Compared with DMSO group,50％ MWT was significantly increased and accompanied with the decrease of protein p-p38,IL-18,IL-lgR and IL-18 mRNA in Ost group after drug administration (P ＜ 0.05).The correlation analysis between protein concentration of p38 MAPK and IL-18 mRNA showed that the Spearman correlation coefficient was 0.9 (P ＜ 0.05).Conclusion p-p38 and IL-18 of spinal dorsal horn participate in the rat model with inflammatory radicular pain induced by nucleus pulposus,and IL-18R plays a role in maintenance of the pain.Osthole administered epidurally in the early stage of pain could alleviate the pain for a long time,which may be related with inhibiting p38 MAPK signaling related pathways.

OBJECTIVETo examine the long-term outcomes of total colonic aganglionosis (TCA) and to evaluate their nutritional status.

METHODSEleven pediatric patients treated for TCA between January 1999 and December 2010 were included in the study and followed up. Physical measurements including height, weight and laboratory tests were assessed. Anorectal functions were evaluated with Kelly score and quality of life(QOL) using questionnaire.

RESULTSThe length of follow-up ranged from 8 to 147 months. The children had satisfactory anorectal function (Kelly score, 5-6). One child had a Kelly score of 3. The children who were followed up less than 48 months had significant higher Kelly scores compared with those with more than 48 months follow-up(P<0.05). QOL was good in nine patients (QOL score, 9-10) and moderate (score, 7-8) in 2 patients. Weight-for-age was normal in 2 patients, mild malnutrition in 6 patients, and moderate malnutrition in 3 patients. Height-for-age was normal in 6 patients, mild malnutrition in 3 patients, and moderate malnutrition in 2 patients. The serum albumin was(49.0±2.7) g/L in children with well-educated parents, significantly higher than those with poorly-educated parents(44.3±1.9) g/L(P<0.05).

CONCLUSIONSLong-term outcomes of children with TCA are satisfactory with good anorectal function and quality of life. Low body weight is more common than low height. Children with well-educated parents have better nutrition status.

Follow-Up Studies ; Hirschsprung Disease ; surgery ; Humans ; Infant ; Male ; Nutritional Status ; Treatment Outcome

OBJECTIVETo investigate the effect of sumoylation of alpha-synuclein by SUMO-1 on the mitochondria subcellular localization of alpha-synuclein and its degradation via ubiquitin-proteasome system.

METHODSPrimers of wild-type, A53T pathogenic mutant and K96R mutant of human alpha-synuclein were designed to amplify the corresponding cDNAs without stop codon. The cDNAs were cloned into pGEM T-easy vector, analyzed by using enzyme mapping and DNA sequencing, and subcloned into pEGFP-N1 vector. The recombinant plasmids of pEGFP-alpha-synuclein-WT, pEGFP-alpha-synuclein-A53T and pEGFP-alpha-synuclein-K96R were transfected into HEK293 cells by lipofectamine method. The expression of the alpha-synuclein protein was measured by immunofluorescence and confocal microscope. Then mitochondria staining as well as immunofluorescence were utilized to investigate the effect of wild-type, A53T mutant and sumoylation of alpha-synuclein on mitochondria subcellular localization of alpha-synuclein. The effect of sumoylation of alpha-synuclein on its degradation via the ubiquitin-proteasome system in the cells was assayed by Western-blot.

RESULTSThe enzyme mapping suggested that the eukaryotic expression plasmids for human wild-type, A53T and K96R mutants of the alpha-synuclein gene were constructed successfully. By immunofluorescence and confocal microscope, it was observed that alpha-synuclein-WT and alpha-synuclein-A53T proteins aggregated in cytoplasm, and alpha-synuclein-K96R protein aggregation was decreased in cytoplasm of cultured cells. The alpha-synuclein proteins of wild-type, A53T and K96R mutants were co-localized with mitochondria. Western-blot analysis revealed that both wild-type and A53T mutant affected the amount of the ubiquitinated proteins.

CONCLUSIONNeither overexpression of wild-type and A53T pathogenic mutant alpha-synuclein, nor sumoylation of alpha-synuclein, affected the subcellular localization in the mitochondria. However, overexpression of wild-type and A53T mutant alpha-synuclein affected the amount of the ubiquitinated proteins.

Blotting, Western ; Cell Line ; Humans ; Mitochondria ; metabolism ; Proteasome Endopeptidase Complex ; metabolism ; SUMO-1 Protein ; metabolism ; Ubiquitin ; metabolism ; alpha-Synuclein ; metabolism

Adolescent ; Adult ; Female ; Humans ; Male ; Mercury ; urine ; Middle Aged ; Reference Values ; Spectrophotometry, Atomic

The objective of this study was to identify the frequency and types of JAK2V617F mutation in chinese patients with essential thrombocythemia (ET), to quantitatively detect the level of mutation transcripts and to investigate its clinical significance. The frequency and types of JAK2V617F mutation were detected by amplification-refractory mutation sequencing polymerase chain reaction (ARMS-PCR), the transcript level of JAK2V617F mutation was determined by using capillary electrophoresis. The results indicated that the JAK2V617F mutation was detected in 59 out of 98 patient with ET, 18 of whom were homozygous mutation. The mean age of patients with homozygous and heterozygous mutation was higher than that of patients with wild type mutation (p < 0.05). The quantitative assay using capillary electrophoresis showed that the transcript level of JAK2V617F mutation in patients with homozygous mutation was (89.9 +/- 6.7)%, which was higher than that in patients with heterozygous mutation (57.1 +/- 6.7)% (p < 0.05); the transcript level of JAK2V617F mutation in patients with age < 60 years was (62.3 +/- 16.5)%, which was lower than that in patients with age > 60 years (72.4% +/- 15.8)% (p < 0.05). The rate of thrombotic complications in patients with JAK2V617F-positive was higher than that in patients with JAK2V617F-negative in which the rate of thrombotic complication in patients with homozygous mutation was higher than that in patients with heterozygous mutation (p < 0.05). Compared with patients without thrombotic events, there were higher level of transcripts of JAK2V617F mutation in patients with thrombotic events. It is concluded that the JAK2V617F positive and negative patients with ET display the different clinical features, therefore, the analysis of mutation types and detection of transcript levels not only helps to identify the disease status and progression, but also guides the treatment of ET patients.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Genotype ; Humans ; Janus Kinase 2 ; genetics ; Male ; Middle Aged ; Mutation ; Thrombocythemia, Essential ; genetics ; pathology ; Young Adult

OBJECTIVETo investigate the frequency and mutational status of JAK2 mutation in Chinese patients with chronic myeloproliferative neoplasms (MPN) and study the relative quantitation and clinical implications of mutated JAK2 transcript.

METHODSJAK2 mutation and the mutational status were screened with amplification-refractory mutation sequencing polymerase chain reaction (ARMS-PCR), the relative quantity of mutated JAK2 mRNA by using capillary electrophoresis.

RESULTSJAK2V617F mutation was detected in 95 of 135 MPN patients, including 37 (97.4%) of 38 polycythemia vera (PV), 56 (59.6%) of 94 essential thrombocythemia (ET) and 2 of 3 idiopathic myelofibrosis (IMF) patients; the difference between the mutations in PV and ET was significant (P<0.05). Of 95 JAK2V617F patients examined, 18/38 PV patients (47.3%) and 17/94 (18.1%) ET patients and 1 IMF patient were homozygotes, and ET patients showed lower prevalence of homozygote (P<0.05). In 95 MPN patients, the mutated mRNA ratio was higher in homozygote than in heterozygote patients. PV heterozygote patients showed higher levels of mutated JAK2 mRNA than ET heterozygote patients (P<0.05). The levels of JAK2V617F mRNA in patients over 60 years of age were significantly higher than that in those less than 60 years of age (P<0.001). Higher leukocyte counts were observed in PV and ET patients with higher levels of mutated JAK2 mRNA (P<0.05). The presence of JAK2V617F was found to be significantly associated with higher hemoglobin level in ET patients. Cytogenetic analysis was performed in 101 of the 135 patients, the association between abnormal karyotype and JAK2V617F was not found.

CONCLUSIONThe ARMS-PCR technique can be used to detect the frequency and mutational status of JAK2V617F mutation, and along with capillary electrophoresis, the estimation of minimal residual disease becomes possible.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; DNA Mutational Analysis ; Electrophoresis, Capillary ; Female ; Humans ; Janus Kinase 2 ; genetics ; Male ; Middle Aged ; Mutation ; Myeloproliferative Disorders ; genetics ; Polymerase Chain Reaction ; methods ; RNA, Messenger ; genetics ; Young Adult

OBJECTIVETo study the emergency management principles of severe trauma in hospital (injury severity score larger than or equal to 16).

METHODSWe used "ATP principle" to manage severe traumatic patients. The ATP principle is composed of: 1) attending surgeons offering initial management (A); 2) teamwork commencement immediately after patients admitted to hospital (T); 3) parallel principle, ie, emergency resuscitation, evaluation and laboratory test performed simultaneously (P). Clinical effects before and after applying ATP principle were retrospectively analyzed and compared.

RESULTSDuring January 1, 2002 to December 31, 2003, 338 patients were treated without applying ATP principle, in which ISS was 25.9+/-6.4, 152 cases died with the mortality being 39.2%, and the time stayed in emergency department and the time to operation room after admission were (102.8+/-16.7) min, (140.3+/-20.6) min, respectively. During January 1, 2004 to December 31, 2005, 438 patients were treated based on ATP principle, in which ISS was 28.6+/-7.8, 87 cases died with the mortality being 19.9%, and the time in emergency department and the time to operation room after admission were (69.5+/-11.5) min, (89.6+/-9.3) min, respectively. ISS showed no significant difference between the two groups (P larger than 0.05) but the mortality, the time stayed in emergency department and the time to operation room after admission were greatly reduced and showed significant difference between the two groups (P less than 0.05).

CONCLUSIONSApplying ATP principle to treat severe traumatic patients can shorten emergency treatment time in hospital and decrease mortality.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; China ; Emergency Service, Hospital ; organization & administration ; Female ; Humans ; Injury Severity Score ; Male ; Middle Aged ; Patient Care Team ; Retrospective Studies ; Triage ; Wounds and Injuries ; classification ; mortality ; therapy