[Evaluation of the symmetry in nasolabial area of unilateral complete cleft lip with the method of rotation descent step by step].

PURPOSE: To evaluate the symmetry and stability of nasolabial area of unilateral complete cleft lip (UCCL) after primary repair with the method of rotation descent step by step. METHODS: Thirty patients with UCCL who were operated on were photographed before, 1 week after and 1 year after surgery, the distances from alare point (al), sub alare point (sba), christa phlitri point (cph) and chelion point (ch) to the facial vertical midline (VML) were measured and compared with the opposite side by paired t test. Statistical analysis was performed with SPSS 19.0 software package. RESULTS: There were significant differences in al, sba and cph between the cleft side and non-cleft side before(P<0.05) and 1 week after surgery; significant differences were found in al, sba and ch between the affected and unaffected side (P<0.05); but 1 year after surgery, there was no significant difference between the two sides except sba. After surgery, all the distances from VML were less than those before surgery. There was no significant difference in symmetry rate between 1 week and 1 year after surgery for all the points except sba. CONCLUSIONS: The results indicated that the method of rotation descent step by step is very helpful in reconstruction of lip symmetry, but primary repair can not achieve full recovery of nasal symmetry.

Regulation of hippocampal neuronal apoptosis and autophagy in mice with sepsis-associated encephalopathy by immunity-related GTPase M1.

AIMS: Sepsis-associated encephalopathy (SAE) is a common complication of severe sepsis. Our goal was to investigate the role of immunity-related GTPase M1 (IRGM1) in SAE and its underlying mechanism. METHODS: A mouse sepsis model was established by cecal ligation and perforation. SAE was diagnosed by behavior, electroencephalography, and somatosensory evoked potentials. Wild-type mice with SAE were treated with SB203580 to block the p38 mitogen-activated protein kinase (MAPK) signaling pathway. We assessed hippocampal histological changes and the expression of IRGM1, interferon-γ (IFN-γ), and p38 MAPK signaling pathway-related proteins. RESULTS: Immunity-related GTPase M1 and IFN-γ levels increased in the hippocampus, with apoptosis, autophagy, and the p38 MAPK signaling pathway activated in neurons. Administration of SB203580 to mice with SAE reduced apoptosis and autophagy. Relative to wild-type mice with SAE, the general condition of Irgm1-/- mice with SAE was worsened, the p38 MAPK signaling pathway was inhibited, and neuronal apoptosis and autophagy were reduced. The absence of IRGM1 exacerbated SAE, with higher p38 MAPK signaling pathway activity and increased apoptosis and autophagy. CONCLUSIONS: During SAE, IRGM1 can at least partially regulate apoptosis and autophagy in hippocampal neurons through the p38 MAPK signaling pathway.

[Role of p38 Mitogen-activated Protein Kinase Signaling Pathway in the Hippocampal Neurons Autophagy of Rats with Sepsis].

OBJECTIVE: To investigate the role of p38 mitogen-activated protein kinase (MAPK) signaling pathway in autophagy of neurons in hippocampus of sepsis rats. METHODS: A sepsis model was established by cecal ligation and puncture (CLP). SD rats were randomly divided into sham-operated group (sham group), model group (CLP group), vehicle-treated group (CLP+Veh group) and inhibitor-treated group (CLP+SB203580 group), and each group was divided into 3, 6, 12, 24 and 48 h subgroups. CLP+Veh group and CLP+SB203580 group were injected with 1% DMSO 5 µL and 0.1 mmol/L SB203580 5 µL respectively in the lateral ventricle, and CLP was established 30 min after injection. The sham group only turned over the cecum and closed the abdomen without other treatments. The vital signs of rats were monitored, including mean arterial pressure (MAP) and heart rate (HR). Neurobehavioral score was used to investigate the brain injury in rats. Histopathological changes in hippocampus of rats were observed by HE staining. The process of neuronal autophagy in hippocampal of rats was observed under transmission electron microscope (TEM). Western blot assay was performed to detect the expression of microtubule associated protein 1 light chain 3 (LC3)Ⅱ, LC3Ⅰ, selective autophagy adaptor protein p62/sequestosome-1 (p62/SQSTM1), MAPK-activated protein kinase 2 (MK-2) and phosphorylation MK-2 (p-MK-2) in the hippocampus. The expressions of LC3 and p62/SQSTM1 in hippocampal neurons of rats were observed by immunofluorescence. RESULTS: At different time points, MAP of CLP group was lower than sham group, while HR was higher than sham group, the change was most obvious at 12 h after molding; the neurobehavioral score of CLP group was the lowest; the histopathological changes in the hippocampus were obvious; and many autophagy vacuoles were observed under transmission electron microscope; compared with CLP group, the neurobehavioral score of CLP+SB203580 group increased; the pathological changes in the hippocampus improved; the inclusions in autophagy vacuoles were degraded under transmission electron microscopy; Western blot results showed:compared with sham group, expression of-LC3Ⅱ/LC3Ⅰ, p-MK-2/MK-2 increased, and p62/SQSTM1 decreased in hippocampal tissue of CLP group in rat, the former reaches its peak at 12 h, the latter bottomed out at 12 h. Compared with the other groups, at 12 h of modeling, the expression of LC3Ⅱ/LC3Ⅰ, p-MK-2/MK-2 was further increased, the expression of p62/SQSTM1 decreased further in hippocampal tissue of CLP+SB203580 group in rat (P < 0.05); immunofluorescence observation showed that localization and expression of LC3 and p62/SQSTM1 in NeuN were consistent with Western blot. CONCLUSION: Inhibition of p38 MAPK signaling pathway in sepsis rats can further activate autophagy and protect neurons in the hippocampus.

Low-dose anisomycin sensitizes glucocorticoid-resistant T-acute lymphoblastic leukemia CEM-C1 cells to dexamethasone-induced apoptosis through activation of glucocorticoid receptor and p38-MAPK/JNK.

Glucocorticoid (GC) resistance in children with acute lymphoblastic leukemia (ALL) usually resulted in the failure of treatment. Exploring new agents to overcome GC resistance is important. Here we reported for the first time that low-dose anisomycin has the potential to sensitize GC-resistant T-ALL CEM-C1 cells to dexamethasone (DEX). Compared with the use of DEX or low-dose anisomycin alone, co-treatment with them resulted in a significant increase of growth inhibition, apoptosis and cell cycle arrest in CEM-C1 cells through induction of activated caspase-3 and up-regulation of Bim, p21and p27, and down-regulation of Mcl-1, Bcl-2, c-myc, cyclin A and cyclin D1. Furthermore, co-treatment remarkably activated glucocorticoid receptor (GR), p38-MAPK and JNK, and all of them were canceled only by the GR inhibitor RU486, indicating GR might be an at the upstream of GR-p38-MAPK/JNK pathway. We conclude that low-dose anisomycin sensitizes GC-resistant CEM-C1 cells to DEX and this effect is mediated, at least in part, by activation of the GR-p38-MAPK/JNK signaling pathway.

Differential expression of anthocyanin biosynthetic genes and transcription factor PcMYB10 in pears (Pyrus communis L.).

Anthocyanin biosynthesis in various plants is affected by environmental conditions and controlled by the transcription level of the corresponding genes. In pears (Pyrus communis cv. 'Wujiuxiang'), anthocyanin biosynthesis is significantly induced during low temperature storage compared with that at room temperature. We further examined the transcriptional levels of anthocyanin biosynthetic genes in 'Wujiuxiang' pears during developmental ripening and temperature-induced storage. The expression of genes that encode flavanone 3-hydroxylase, dihydroflavonol 4-reductase, anthocyanidin synthase, UDP-glucose: flavonoid 3-O-glucosyltransferase, and R2R3 MYB transcription factor (PcMYB10) was strongly positively correlated with anthocyanin accumulation in 'Wujiuxiang' pears in response to both developmental and cold-temperature induction. Hierarchical clustering analysis revealed the expression patterns of the set of target genes, of which PcMYB10 and most anthocyanin biosynthetic genes were related to the same cluster. The present work may help explore the molecular mechanism that regulates anthocyanin biosynthesis and its response to abiotic stress at the transcriptional level in plants.

[Risk factors of heart and lung failure in children with severe hand, foot and mouth disease and treatment experience].

OBJECTIVE: To study risk factors for severe hand, foot and mouth disease (HFMD) complicated by heart and lung failure and treatment experience. METHODS: A total of 198 children with severe HFMD between March and August in 2011 were enrolled. Univariate analysis and logistic regression model were used to analyze the risk factors severe HFMD complicated by heart and lung failure. The effects of combination therapy with immunoglobulin+dexamethasone+ribavirin were observed. RESULTS: Univariate analysis indicated that HFMD patients with heart and lung failure had higher proportions of consciousness, tachypnoea, abnormal hemodynamics, increased troponin and EV71 infection than HFMD patients without heart and lung failure (P<0.05).Multivariate logistic regression analysis indicated that tachypnoea, abnormal hemodynamics and EV71 infection were the main risk factors for heart and lung failure. Compared with combination therapy with dexamethasone+ribavirin, combination therapy with immunoglobulin+dexamethasone+ribavirin was more effective for preventing hemodynamic changes in children with severe HFMD (P<0.01). Compared with HFMD patients with heart and lung failure, the effect of the combination therapy with immunoglobulin+dexamethasone+ribavirin was better in HFMD patients without heart and lung failure (P<0.01). CONCLUSIONS: The main risk factors for heart and lung failure in children with severe HFMD include tachypnoea, abnormal hemodynamics and EV71 infection. Early combination therapy with immunoglobulin+dexamethasone+ribavirin can reduce the incidence of heart and lung failure in children with severe HFMD.

Peripherally inserted central catheters and the incidence of candidal sepsis in VLBW and ELBW infants: is sepsis increased?

BACKGROUND: Peripherally inserted central catheters (PICCs) have been widely used in neonatal clinics. However, the complications such as infection after PICC treatment are also confronting neonatologists especially in developing countries. This study was undertaken to investigate whether PICCs is a safe treatment for very low birth weight (VLBW) infants and extremely low birth weight (ELBW) infants. METHODS: Fifty-nine VLBW and ELBW infants receiving PICCs and 89 VLBW and ELBW infants receiving peripheral intravenous catheters (PIVCs) were included in this study. The incidence of sepsis and mortality were compared retrospectively between the two groups. RESULTS: There was no difference in the total sepsis incidence and mortality between the PICCs and PIVCs groups (P=0.11 and P=0.61 respectively). However, the candidal sepsis incidence was higher in the PICCs group than in the PIVCs group [6/59 (10.2%) vs 2/89 (2.2%); P=0.044 (Exat Sig. 1-sided), OR=4.93, 95% CI 0.96-25.3]. CONCLUSION: Placement and indwelling of PICCs are a potential risk factor for candidal sepsis among VLBW and ELBW infants.

[hTERT gene expression in children with beta-thalassemia major].

OBJECTIVE: Human telomerase reverse transcriptase (hTERT) is a rate-limiting enzyme which dictates the activity of human telomerase and thus decides the life span of cells. The aim of this study was to explore the expression of hTERT in bone marrow from children with beta-thalassemia major and the relationship between the expression of hTERT and hemoglobin levels. METHODS: Multiple allele specific polymerase chain reaction (MASPCR) was used for targeted DNA amplification and gene mutation analysis of beta-thalassemia. hTERT mRNA expression in bone marrow was examined using real-time reverse transcription polymerase chain reaction (RT-PCR) analysis in 29 children with beta-thalassemia major, in 10 children with agranulocytosis and in K562 cell line. The hemoglobin levels in peripheral blood were measured. The relationship between hTERT expression and hemoglobin levels was evaluated by the Spearman test in the beta-thalassemia major group. RESULTS: hTERT mRNA expression significantly increased in bone marrow from children with beta-thalassemia major compared with that from children with agranulocytosis (0.2928+/- 0.0838 vs 0.0993+/- 0.0336; P<0.01), but was significantly lower than that in K562 cell line (0.8291+/- 0.0908) (P<0.01). A significantly inverse correlation was found between hTERT mRNA expression and hemoglobin levels (r=-0.841, P<0.01). CONCLUSIONS: A low hemoglobin concentration might contribute to the up-regulation of marrow hTERT expression in children with beta-thalassemia major.

[Administration of low-dose heparin in total nutrient admixture prevents central venous catheter-related infections in neonates].

OBJECTIVE: Peripherally inserted central catheter (PICC) is widely used to provide a long-term access for the administration of total parenteral nutrition and medications. Catheter-related infections (CRI) are common complications of PICC. The purpose of this retrospective study was to investigate the role of low-dose heparin added to the total nutrient admixture (CTNA) in the prevention of CRI. METHODS: Eighty-three neonates who underwent PICC received TNA with (heparin group, n=43) or without heparin (0.5 U/mL) (control group, n=40). The incidence of CRI was compared between the two groups. RESULTS: The incidences of catheter obstruction (5% vs 20%) and the catheter-tip colonization (2% vs 18%) in the heparin group were significantly lower than those in the control group (p<0.05). None of the neonates in the heparin group had clinical evidence of catheter-related bloodstream infection, but 5 cases in the control group (p<0.05). CONCLUSIONS: The administration of low-dose heparin in TNA may decrease the incidences of catheter obstruction and CRI.

[Comparative study on clinical features between TEL-AML1 positive and negative childhood acute lymphoblastic leukemia].

OBJECTIVE: To determine the incidence of TEL-AML1 fusion gene in childhood acute lymphoblastic leukemia (ALL) and to compare the clinical features between TEL-AML1 positive and negative patients. METHODS: Samples of bone marrow or peripheral blood were collected from 95 newly diagnosed ALL children and the TEL-AML1 fusion gene was detected using nested reverse transcription-polymerase chain reaction (RT-PCR). The ALL patients were stratified into TEL-AML1 positive and negative groups and the clinical features were compared. RESULTS: Among 95 patients, 20 (21.05%) were TEL-AML1 positive. The median age of TEL-AML1 positive patients was 5.9 years old and M/F ratio was 1.22:1. There were significant differences between TEL-AML1 positive and negative patients in hepatomegaly (2.75 cm vs. 4 cm below costal arch, P=0.006), splenomegaly (0 cm vs. 3 cm below costal arch, P < 0.001), initial white blood cell count (median 7.40 x 10(9)/L vs.18.70 x 10(9)/L, P=0.011), initial peripheral blood blast (median 2.45 x 10(9)/L vs.11.66 x 10(9)/L, P=0.013), hemoglobin level [(61.45 +/- 13.46) g/L vs. (75.89 +/- 23.11) g/L, P=0.003] and serum lactate dehydrogenase [(621.47 +/- 335.85) U/L vs.(1566.64 +/- 1720.45) U/L, P=0.020], while no differences were found between two groups in age, gender ratio, initial platelet count, percentage of blast in bone marrow, immunophenotypes and the expression of myeloid antigen CD13, CD33 and CD34. The prednisone sensitivity test showed that all 12 TEL-AML1 positive patients were good responders, while there were 11 prednisone poor responders among 40 negative patients (27.50%, P < 0.05). Bone marrow examination on day 15 showed no difference in the rate of complete remission between TEL-AML1 positive and negative patients. CONCLUSION: The incidence of TEL-AML1 fusion gene in cases of ALL is 21.05%. The load of leukemia cells in TEL-AML1 positive patients is significantly smaller than its counterparts, and the blast cells in TEL-AML1 positive patients are more sensitive to prednisone, indicating childhood ALL with TEL-AML1 fusion gene has a favorable prognosis.

[Genetic study on 27 children with beta-thalassemia major and their parents in Sichuan area].

OBJECTIVE: This genetic analysis on 27 children with beta-thalassemia major and their parents was performed in an attempt to elucidate the characteristics of gene mutations, and to improve the early diagnosis and prevention of this disease in Sichuan area. METHODS: The hematologic studies of beta-thalassemia included the osmotic fragility of erythrocyte determined by brine osmosis method, the HbF qualitatively determined by one minute anti-alkaline method, the HbA2 assayed by acetyl-cellulose membrane electrophoresis and eluate photometric method, and the total hemoglobin measured by the ferric-cynade method. DNA was extracted from peripheral white blood cell with standard method (phenol-chloroform extract). Multiple allele specific polymerase chain reaction (MASPCR) was used for targeted DNA amplification and gene mutation analysis. RESULTS: It was found that the most common mutants in Sichuan area were CD17 (A-->T), CD41-42 (-TTCT) and IVS II -654 (C-->T). Their percentages were 43.64%, 36.36% and 14.54%, respectively. CONCLUSION: MASPCR method is a simple, effective and inexpensive method for genetic diagnosis of beta-thalassemia major. We identified 3 most common mutants in Sichuan by using MASPCR.

[Preliminary analysis on human telomerase reverse transcriptase gene expression in acute leukemia].

OBJECTIVE: To explore the relationship between gene expression of human telomerase reverse transcriptase (hTERT) and its clinical characteristics in leukemia. METHODS: The protocol of RT-PCR was used to detect the hTERTmRNA expressing levels in peripheral blood samples from leukemic patients under primary treatment(n=42), in complete remission(n=21), with recurrent leukemia (n=4); and from normal subjects (n=5), respectively. RESULTS: The positive percentage of hTERTmRNA expression was 73.81% for the primary treatment cases, and 19.05% for the complete remission cases. All of the recurrent cases gave positive results. One of the normal controls presented low level of hTERTmRNA expression. The expressing level of hTERTmRNA in primary treatment cases was 0.64+/-0.21, in complete remission leukemia 0.31+/-0.16, in recurrent cases 0.84+/-0.09, and in normal controls 0.10. CONCLUSION: The activation of telomerase may be an essential factor in the development of leukemia and usually be the late event in its progression. As an indicator of leukemia cell, the detection of hTERT mRNA may be used in clinical analysis, disease monitoring and prognosis judgement.