β-arrestin 1 Promotes Senescence of Acute Lymphoblastic Leukemia Jurkat Cells / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of β-arrestin1 gene on senescence of T-ALL cells and its possible mechanism.@*METHODS@#The bone marrow specimens of T-ALL patients and controls were collected, the expression of β-arrestin1 and β-arrestin1 in the T-ALL patients was detected by RT-PCR and Western blot, respectively, and the relation of β-arrestin1 expression with the clinical pathologic characteristics and the prognosis of T-ALL patients was analyzed statistically. The stable Jurkat cell line with knocked down or overexpressed β-arrestin1 was constructed, the CCK method was used to detect the Jurkat cell number, the β-gal staining was used to analyze the effect of β-arrestin1 on senescence of Jurkat cells, the cross analysis of RNA-Seg data and KEGG data was performed for screening the possible signaling pathway, and Western blot was performed for varifying the key sites of signaling pathway.@*RESULTS@#The β-arrestin1 expression in specimens of T-ALL patients decreased (P＜0.01), moreover the β-arrestin1 expression negatively related with peripheral blood cell number (r=-0.601), the blasts in peripheral blood (r=-0.516) and extramedullary infiltration (r=-0.359), while positively related with the response to chemotherapy (r=0.393). The detection of stable Jurkat cell line with knocked-down and overexpressed β-arrestin1 found that the β-arrestin 1 could decrease the Jurkat cell number and accelarate the senescence of Jurkat cells (P＜0.05). The cross analysis of RNA-Seg data and KEGG data showed that the senescence of T-ALL cells may be regulated via RAS-P16-PRb-E2F1 by β-arrestin 1. Western bolt confirmed that β-arrestin1 promoted the expression of Ras and p16, and decreased the expression of pRB and E2F1 (P＜0.05).@*CONCLUSIONS@#β-arrestin1 accelerates the senescence of Jurkat cells via Ras-p16-pRb-E2F1, and delays the progression in T-ALL, which may provide a new hypothesis for the pathogenesis of T-ALL.

Screening and verification of key genes in T-cell acute lymphoblastic leukemia / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the key genes in T-cell acute lymphoblastic leukemia (T-ALL) using bioinformatics method to better understand the pathogenic mechanisms of T-ALL.</p><p><b>METHODS</b>The gene expression profiles of GSE14317 were obtained from Gene Expression Omnibus database. The differentially expressed genes (DEGs) in T-ALL were analyzed using R package Limma. The online analysis tool DAVID was used to perform the functional and pathway enrichment analysis. The protein-protein interaction network was constructed by STRING and visualized by Cytoscape. Based on the JASPAR database, the transcription factors (TFs) of the hub genes were obtained. RT-PCR was used to test the mRNA expression level of the key genes.</p><p><b>RESULTS</b>A total of 1443 DEGs were identified, including 800 up-regulated genes and 643 down-regulated genes. These DEGs were significantly enriched in the cell cycle, hematopoietic cell lineage, cytokine-cytokine receptor interaction and T cell receptor signaling pathway. The top 10 hub genes identified from the PPI networks included CDK1, PIK3R1, CCNB1, CCNA2, CDC20, JUN, GNG11, PLK1, PCNA and CCNB2, which were enriched in chemokine signaling pathway, ubiquition mediated proteolysis and cell cycle. In the TF-target gene network, 42 differentially expressed TFs were identified, among which ELF5, HIC2 and MEISI had binding sites with 9 of the candidate hub genes. RT-PCR showed that the mRNA expression level of all the candidate hub genes except for GNG11 were consistent with the gene expression profiles.</p><p><b>CONCLUSION</b>The hub genes CDK1, PIK3R1, CCNB1, CCNA2, CDC20, JUN, PLK1, PCNA, CCNB2, ELF5, HIC2 and MEISI participate in the occurrence of T-ALL. Our finding provides new insights into the pathogenesis of T-ALL.</p>

Heart Rate Variability in Patients with Acute Ischemic Stroke at Different Stages of Renal Dysfunction: A Cross-sectional Observational Study / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Renal function is associated with mortality and functional disabilities in stroke patients, and impaired autonomic function is common in stroke, but little is known regarding its effects on stroke patients with renal dysfunction. This study sought to evaluate the association between autonomic function and stroke in patients with renal dysfunction.</p><p><b>METHODS</b>This study comprised 232 patients with acute ischemic stroke consecutively enrolled from February 2013 to November 2014 at Guangdong Provincial Hospital of Chinese Medicine in China. All patients recruited underwent laboratory evaluation and 24 h Holter electrocardiography (ECG). Autonomic function was measured based on the heart rate variability (HRV) using 24 h Holter ECG. Renal damage was assessed through the estimated glomerular filtration rate (eGFR), and stroke severity was rated according to the National Institutes of Health Stroke Scale (NIHSS). The Barthel index and modified Rankin score were also determined following admission. All the clinical covariates that could potentially affect autonomic outcome variables were adjusted with linear regression.</p><p><b>RESULTS</b>In the patients with a mild or moderate decreased eGFR, the values for the standard deviation of the averaged normal-to-normal RR interval (SDANN) index (P = 0.022), very low frequency (VLF) (P = 0.043), low frequency (LF) (P = 0.023), and ratio of low-to-high frequency power (LF/HF) (P = 0.001) were significantly lower than those in the patients with a normal eGFR. A multinomial linear regression indicated that eGFR (t = 2.47, P = 0.014), gender (t = -3.60, P < 0.001), and a history of hypertension (t = -2.65, P = 0.008) were the risk factors of LF/HF; the NIHSS score (SDANN index: t = -3.83, P < 0.001; VLF: t = -3.07, P = 0.002; LF: t = -2.79, P = 0.006) and a history of diabetes (SDANN index: t = -3.58, P < 0.001; VLF: t = -2.54, P = 0.012; LF: t = -2.87, P = 0.004) were independent factors for the SDANN index, VLF, and LF; the Oxfordshire Community Stroke Project (t = -2.38, P = 0.018) was related to the SDANN index.</p><p><b>CONCLUSIONS</b>Autonomic dysfunction is aggravated with the progression of eGFR stage in patients with acute ischemic stroke; the eGFR is an independent factor of LF/HF in the adjusted models. Stroke severity and a history of diabetes are more significantly associated with HRV in patients with acute ischemic stroke at different stages of renal dysfunction.</p>

TGF-β1-induced synthesis of collagen fibers in skeletal muscle-derived stem cells / 华中科技大学学报（医学）（英德文版）

The aim of this study was to investigate the mechanism of deposition of extracellular matrix induced by TGF-β1 in skeletal muscle-derived stem cells (MDSCs). Rat skeletal MDSCs were obtained by using preplate technique, and divided into four groups: group A (control group), group B (treated with TGF-β1, 10 ng/mL), group C (treated with TGF-β1 and anti-connective tissue growth factor (CTGF), both in 10 ng/mL), and group D (treated with anti-CTGF, 10 ng/mL). The expression of CTGF, collagen type-I (COL-I) and collagen type-III (COL-III) in MDSCs was examined by using RT-PCR, Western blot and immunofluorescent stain. It was found that one day after TGF-β1 treatment, the expression of CTGF, COL-I and COL-III was increased dramatically. CTGF expression reached the peak on the day 2, and then decreased rapidly to a level of control group on the day 5. COL-I and COL-III mRNA levels were overexpresed on the day 2 and 3 respectively, while their protein expression levels were up-regulated on the day 2 and reached the peak on the day 7. In group C, anti-CTGF could partly suppress the overexpression of COL-I and COL-II induced by TGF-β1 one day after adding CTGF antibody. It was concluded that TGF-β1 could induce MDSCs to express CTGF, and promote the production of COL-I and COL-III. In contrast, CTGF antibody could partially inhibit the effect of TGF-β1 on the MDSCs by reducing the expression of COL-I and COL-III. Taken together, we demonstrated that TGF-β1-CTGF signaling played a crucial role in MDSCs synthesizing collagen proteins in vitro, which provided theoretical basis for exploring the methods postponing skeletal muscle fibrosis after nerve injury.

TGF-β1-induced synthesis of collagen fibers in skeletal muscle-derived stem cells / 华中科技大学学报（医学）（英德文版）

The aim of this study was to investigate the mechanism of deposition of extracellular matrix induced by TGF-β1 in skeletal muscle-derived stem cells (MDSCs). Rat skeletal MDSCs were obtained by using preplate technique, and divided into four groups: group A (control group), group B (treated with TGF-β1, 10 ng/mL), group C (treated with TGF-β1 and anti-connective tissue growth factor (CTGF), both in 10 ng/mL), and group D (treated with anti-CTGF, 10 ng/mL). The expression of CTGF, collagen type-I (COL-I) and collagen type-III (COL-III) in MDSCs was examined by using RT-PCR, Western blot and immunofluorescent stain. It was found that one day after TGF-β1 treatment, the expression of CTGF, COL-I and COL-III was increased dramatically. CTGF expression reached the peak on the day 2, and then decreased rapidly to a level of control group on the day 5. COL-I and COL-III mRNA levels were overexpresed on the day 2 and 3 respectively, while their protein expression levels were up-regulated on the day 2 and reached the peak on the day 7. In group C, anti-CTGF could partly suppress the overexpression of COL-I and COL-II induced by TGF-β1 one day after adding CTGF antibody. It was concluded that TGF-β1 could induce MDSCs to express CTGF, and promote the production of COL-I and COL-III. In contrast, CTGF antibody could partially inhibit the effect of TGF-β1 on the MDSCs by reducing the expression of COL-I and COL-III. Taken together, we demonstrated that TGF-β1-CTGF signaling played a crucial role in MDSCs synthesizing collagen proteins in vitro, which provided theoretical basis for exploring the methods postponing skeletal muscle fibrosis after nerve injury.

Brain arousal dysfunction in severe craniocerebral injury treated with acupuncture / 中国针灸

<p><b>OBJECTIVE</b>To explore the early rehabilitation effect of acupuncture on brain arousal in severe craniocerebral injury.</p><p><b>METHODS</b>One hundred and two cases of severe craniocerebral injury were randomly divided into an observation group and a control group, 51 cases in each one. Based on the conventional nursing care in neurological external medicine, in observation group, acupuncture was applied at Shuigou (GV 26), Neiguan (PC 6) and Sanyinjiao (SP 6) mainly. In control group, functional electric stimulation was applied at stimulate the affected muscles of the upper limbs. Thirty days later, the lucid rate from coma, lucid interval and clinical efficacy were compared between two groups. RESULTS; The lucid rate from coma was 82.4% (42/51) in observation group, which was higher than 56.9% (29/51) in control group (P < 0.01). The lucid interval in observation group was shortened remarkably as compared with control group (P < 0.01), and the clinical efficacy was superior apparently to that in control group (P < 0.01).</p><p><b>CONCLUSION</b>On the basis of conventional treatment, acupuncture intervention at early stage can accelerate the recovery of brain arousal function in patients with severe craniocerebral injury.</p>

Effect of intrauterine hepatitis B virus infection on peripheral blood mononuclear cells interferon-gamma and interleukin-4 in newborns / 中华儿科杂志

<p><b>OBJECTIVE</b>To observe the effect of intrauterine hepatitis B virus (HBV) infection on peripheral blood mononuclear cells function of secreting interferon-gamma and interleukin-4.</p><p><b>METHODS</b>Pregnant women were systematically screened for HBsAg and HBeAg when attending the antenatal clinic at the Qinhuangdao Maternal and Child Health Hospital. Totally 67 pairs of mothers and infants were enrolled into this study after obtaining the women's consent. Venous blood samples were collected from the infants within 6 hours after birth and before HBIG injection and HBVac immunization. Blood sample was taken from the mother at or after the time when the infant was born. HBV DNA in plasma and PBMC from mothers and their newborns were examined using polymerase chain reaction (PCR). According to HBV DNA in PBMC of newborns, they were divided into two groups. The PBMCs isolated from newborn were cultured with purified HBsAg or phytohemagglutinin (PHA). The supernatant interleukin-4 and interferon-gamma level was measured by using enzyme linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>In 19 newborns PBMC was positive for HBV DNA. Maternal PBMC HBV DNA positivity was associated with high rate of intrauterine HBV infection in the infants (chi2 = 7.58, P < 0.01). Compared with the infants whose PBMC HBV DNA was negative, the infants with PBMC positive for HBV DNA expressed a lower level interferon-gamma secretion after purified HBsAg stimulation (t = 4.71, P < 0.01), however, no significant difference was seen after PHA stimulation (t = 1.21, P > 0.05). The supernatant IL-4 level detected after stimulation with purified HBsAg was higher in the newborns whose PBMC HBV DNA was positive as compared with those negative for PBMC HBV DNA (t = -8.51, P < 0.05). The level of IL-4 did not show any significant difference after stimulation with PHA between the PBMC HBV DNA negative and positive groups (t = -2.40, P > 0.05).</p><p><b>CONCLUSION</b>Infection with HBV of maternal PBMC is responsible for perinatal newborn's PBMC HBV infection and it may be an important route of HBV vertical transmission. Infants whose mothers were positive for HBsAg, HBeAg and HBV DNA were at extraordinarily high risk for hepatitis B virus infection. PBMC infected with HBV could influence the status of humoral and cellular immunity resulting in persistent HBV infection and recurrent mother to infant transmission of HBV. Low responses of interferon-gamma and high interleukin-4 transcription upon specific stimulation exist in infants whose PBMC were positive for HBV DNA in uterus may contribute to immune tolerance to HBV.</p>

Association of Q576R polymorphism in the interleukin-4 receptor gene with serum IgE levels in children with asthma / 中国当代儿科杂志

<p><b>OBJECTIVE</b>Interleukin-4 plays a key role in the development of asthma. Overseas studies have shown that Q576R polymorphism in the interleukin-4 receptor (IL-4R) gene is related to asthma as well as increased serum IgE levels. This study was designed to investigate the association of Q576R polymorphism in IL-4R gene with childhood asthma and serum IgE levels.</p><p><b>METHODS</b>The polymorphism of IL-4R Q576R was determined by PCR/RFLP and serum total IgE level was measured using ELISA in 94 children with asthma. Sixty-eight healthy children served as controls.</p><p><b>RESULTS</b>The distribution frequency of heterozygous genotype Q576R (41%) and mutant allele R576 (26%) was significantly higher in children with asthma than that of controls (16% each) (P < 0.01; P < 0.05). The total serum IgE level between patients with genotype Q576R and Q576Q was not significantly different (225.78 +/- 51.43 IU/mL vs 163.24 +/- 31.32 IU/mL, P> 0.05).</p><p><b>CONCLUSIONS</b>The mutant R576 allele of IL-4R may be one of the candidate genes for susceptibility to asthma. Allele R576 of IL-4R is related to asthma but is irrelevant to the total serum IgE level in children with asthma.</p>

Level of vitamin K-dependent coagulation factors in premature infants and the influence of maternal antenatal administration of vitamin K1 on their activity / 中华儿科杂志

<p><b>OBJECTIVE</b>Infants less than 35 weeks of gestational age are susceptible to peri-/intraventricular hemorrhage (PIVH). This may be due in part to low concentrations of vitamin K-dependent coagulation factors. This study was conducted to determine the umbilical cord blood activities of vitamin K-dependent coagulation factors II, VII, IX and X in premature infants to understand whether preterm infants have absence status of these factors the changes of theses factors' activities in premature infants' umbilical blood after vitamin K(1) was given to mothers antenatally and the preventing effectiveness of PIVH after maternal antenatal supplement of vitamin K(1).</p><p><b>METHODS</b>Pregnant women in preterm labor at less than 35 weeks of gestational age were randomly selected to receive antenatal vitamin K(1) intramuscular or intravenous injections 10 mg per day for 2 to 7 days (vitamin K(1) group), or no vitamin K(1) treatment (control group). Dexamethone was antenatally given to both groups of pregnant women routinely. Vitamin K(1) group had 44 infants and the control group had 133 infants. During the same period, thirty full-term neonates' cord blood samples were obtained to determine theses factors to compare with those from the premature infants. The cranial ultrasound was performed by a same physician to understand whether the neonates were complicated with PIVH and its severity.</p><p><b>RESULTS</b>The levels of vitamin K-dependent coagulation factors in umbilical blood in control group were significantly lower than those in full-term infants' cord blood (P < 0.05). However, in vitamin K(1) group, supplement of vitamin K(1) antenatally could significantly increase activities of factors II, VII and X in preterm infants' cord blood (P < 0.05). The total occurrence rates of PIVH in vitamin K(1) group and control group were 31.8% and 52.6%, respectively, (P = 0.017), and the frequency of severe PIVH in vitamin K(1) group and control group was 2.3% and 12.0%, respectively (P = 0.057).</p><p><b>CONCLUSION</b>Preterm infants have absence status of vitamin K-dependent coagulation factors. Administration of vitamin K(1) to pregnant women at less than 35 weeks of gestational age resulted in significantly improved activities of vitamin K-dependent coagulation factors II, VII, and X, and a significantly decreased frequency of PIVH and less severe hemorrhage in preterm infants.</p>