Screening Biomarkers of Sudden Coronary Death Based on mRNA Expression Profile of Rat Myocardial Tissues / 法医学杂志

OBJECTIVES@#To explore the differential expression of messenger RNA (mRNA) in myocardial tissues of rats with sudden coronary death (SCD), and to provide ideas for the forensic identification of SCD.@*METHODS@#The rat SCD model was established, and the transcriptome sequencing was performed by next-generation sequencing technology. Differentially expressed genes (DEGs) in myocardial tissues of SCD rats were screened by using the R package limma. A protein-protein interaction (PPI) network was constructed by using the STRING database and Cytoscape 3.8.2 on DEG, and hub genes were screened based on cytoHubba plug-in. Finally, the R package clusterProfiler was used to analyze the biological function and signal pathway enrichment of the selected DEG.@*RESULTS@#A total of 177 DEGs were associated with SCD and were mainly involved in the renin-angiotensin system and PI3K-Akt signaling pathway. The genes including angiotensinogen (AGT), complement component 4a (C4a), Fos proto-oncogene (FOS) and others played key roles in the development of SCD.@*CONCLUSIONS@#Genes such as AGT, C4a, FOS and other genes are expected to be potential biomarkers for forensic identification of SCD. The study based on mRNA expression profile can provide a reference for forensic identification of SCD.

Gene Expression Profiles at Different Time Points after Acute Myocardial Infarction in Mice / 法医学杂志

OBJECTIVES@#To explore the mRNA differential expressions and the sequential change pattern in acute myocardial infarction (AMI) mice.@*METHODS@#The AMI mice relevant dataset GSE4648 was downloaded from Gene Expression Omnibus (GEO). In the dataset, 6 left ventricular myocardial tissue samples were selected at 0.25, 1, 4, 12, 24 and 48 h after operation in AMI group and sham control group, and 6 left ventricular myocardial tissue samples were selected in blank control group, a total of 78 samples were analyzed. Differentially expressed genes (DEGs) were analyzed by R/Bioconductor package limma, functional pathway enrichment analysis was performed by clusterProfiler, protein-protein interaction (PPI) network was constructed by STRING database and Cytoscape software, the key genes were identified by Degree topological algorithm, cluster sequential changes on DEGs were analyzed by Mfuzz.@*RESULTS@#A total of 1 320 DEGs were associated with the development of AMI. Functional enrichment results included cellular catabolic process, regulation of inflammatory response, development of muscle system and vasculature system, cell adhesion and signaling pathways mainly enriched in mitogen-activated protein kinase (MAPK) signaling pathway. The key genes of AMI included MYL7, TSC22D2, HSPA1A, BTG2, NR4A1, RYR2 were up-regulated or down-regulated at 0.25-48 h after the occurrence of AMI.@*CONCLUSIONS@#The functional signaling pathway of DEGs and the sequential expression of key genes in AMI may provide a reference for the forensic identification of AMI.

Feasibility of Hei Xiaoyaosan in Preventing and Treating Alzheimer's Disease Based on Theory of 'Correlation of Liver and Kidney' Based on Ca2+-CaM/CaMKⅡ-CREB Signal Pathway / 中国实验方剂学杂志

Alzheimer's disease（AD） is an incipient aging neurodegenerative disease， which increases rapidly along with the development trend of social aging and seriously threatens the health of the people. In the absence of effective preventive measures， it will have an enormous impact on the socio-economic and healthcare system. The study found that abnormal cell signal transduction is a key link in many diseases. Cell signal transduction theory has been widely used to clarify the essence of traditional Chinese medicine visceral image and the mechanism of traditional Chinese medicine. 'Correlation of Liver and Kidney' is one of the core plates of the theory of 'Correlation of Five Organs'， which is suitable for explaining the pathogenesis of complex diseases and the correlation of multiple syndromes， and guiding the prescription of clinical syndrome. Hei Xiaoyaosan， as the first choice compound for the prevention and treatment of AD based on the theory of "Correlation of Liver and Kidney' in our team， can play the effects of prevention and treatment by soothing liver and nourishing blood， strengthening spleen and tonifying kidney， and promoting brain collaterals and dredging viscerab spirit. Based on the theory of 'Correlation of Liver and Kidney'， this paper expounds the pathogenesis of AD from the perspective of traditional Chinese medicine， and puts forward the methods and ideas of the preventing and treating of AD from Ca2+-calcium/calmodulin dependent protein （CaM）/calcium/calmodulin dependent protein kinaseⅡ（CaMKⅡ）-cyclic adenosine phosphate reactive element binding protein （CREB） cell signal transduction pathway by consulting literatures and previous studies.

Influenza virus activates toll-like receptor 7/nuclear factor-κB signaling pathway to regulate airway inflammatory response in patients with acute exacerbation of chronic obstructive pulmonary diseases / 中华内科杂志

Objective:To explore how influenza A virus (IAV) regulates airway inflammation via activating Toll-like receptor 7(TLR7)/nuclear factor of κB (NF-κB) signaling pathway in patients with acute exacerbation of chronic obstructive pulmonary disease (COPD).Methods:Primary bronchial epithelial cells were isolated and cultured from normal controls and COPD patients. Samples were divided into 6 groups according to different in vitro treatment, including normal epithelial cell group (A), normal cells+IAV group (B), COPD epithelial cell group (C), COPD cells+IAV group (D), normal cells+TLR7 small interference RNA (si-RNA) group (E), COPD cells+TLR7 siRNA group (F). Protein expressions of TLR7 and NF-κB were detected by Western blot after 24h co-culture with IAV and TLR7 siRNA. Interleukin-6 (IL-6) and tumor necrosis factor α (TNF α) were detected by enzyme-linked immunosorbent assay (ELISA).Results:(1) Compared with group A [0.350±0.075 and 0.470±0.034, (53.000±6.532)pg/ml and (17.000±1.625)pg/ml],TLR7, NF-κB protein expression and IL-6, TNF α levels were significantly increased in group B[0.950±0.075 and 1.090±0.078,(185.000±7.874)pg/ml and (32.000±0.838)pg/ml], group C[0.780±0.056 and 0.910±0.045,(138.000±5.100)pg/ml and 29.000±1.323)pg/ml) and group D[1.280±0.031 and 1.540±0.051,(432.000±5.734)pg/ml and (52.000±3.453)pg/ml] (all P<0.01). Compared with group C TLR7, NF-κB protein expression and IL-6, TNF α levels were significantly increased in group D ( P<0.01). (2) Compared with the group A[0.530±0.023 and 0.800±0.046,(51.000±0.327)pg/ml and (14.000±0.314)pg/ml], TLR7, NF-κB protein expression and IL-6, TNF α levels were significantly decreased in the group E[0.350±0.047 and 0.510±0.067,(26.000±1.081)pg/ml and(8.000±0.526)pg/ml] ( P<0.05). Compared with group C[1.080±0.078 and 1.280±0.034,(125.000±2.249)pg/ml and (28.000±1.010)pg/ml], TLR7, NF-κB protein expression and IL-6, TNF α levels decreased in the group F[0.880±0.056 and 1.040±0.029,(83.000±1.125)pg/ml and (21.000±0.429)pg/ml] ( P<0.05). Conclusion:Influenza viruses activate TLR7/NF-κB signaling pathway to regulate airway inflammation storms in patients with acute exacerbation of COPD. New therapeutic targets of acute exacerbation COPD may be studied based on these inflammation responses to influenza viruses.

Intracranial atherosclerotic plaque enhancement associated with ischemic stroke: a study with three-dimensional high-resolution magnetic resonance vessel wall imaging / 复旦学报（医学版）

Objective To evaluate the reproducibility of three-dimensional high-resolution magnetic resonance imaging (HR-MRI) for vessel wall in demonstration of intracranial atherosclerotic plaque enhancement and to explore the relationship between plaque enhancement and ischemic stroke.Methods Fifty-two patients with ischemic stroke underwent traditional head MRI,three-dimensional time of flight magnetic resonance angiography and HR-MRI on a 3.0 T MRI scanner.Each identified intracranial plaque was classified as either culprit (the only or most stenotic lesion upstream from a stroke) or non-culprit (not the most stenotic lesion upstream from a stroke or not within the vascular territory of a stroke).The degree of plaque enhancement was graded by two independent radiologists.The degree of plaque enhancement and luminal stenosis were compared between the culprit group and the non-culprit group by using Mann-Whitney U test.Binary logistic regression analysis was performed to assess the relation between the degree of plaque enhancement and culprit plaques.Results Total 118 plaques were identified in 52 patients with ischemic stroke (52 culprit plaques and 66 non-culprit plaques).The degree of enhancement was rated as strong,moderate and none in 40,9 and 3 culprit plaques,and in 4,24 and 38 non-culprit plaques.Both intra-observer and inter-observer agreement were high for identification of plaque enhancement (kappa＞ 0.75).For culprit plaques group,the degree of plaque enhancement(Z =-7.787,P＜0.01) and luminal stenosis (Z =-5.327,P＜0.01) were significantly higher than those in the non-culprit group.Binary logistic regression analysis revealed that strong enhancement of plaques was independently associated with culprit plaques (OR:74.3,95％CI:15.0-367.1,P＜0.01).Conclusions Three-dimensional HR-MRI detects enhancement of intracranial plaques with high reproducibility.Enhancement is more common in culprit plaques and is associated with the likelihood of ischemic stroke.

Taraxerol induces cell apoptosis through a mitochondria-mediated pathway in HeLa cells

Objective: Taraxerol acetate has potent anti-cancer effects via the induction of apoptosis, autophagy, cell cycle arrest, and inhibition of cell migration. However, whether taraxerol induced apoptosis and its underlying mechanisms of action is not clear. In the present study, we assess the effects of taraxerol on the mitochondrial apoptotic pathway and determine the release of cytochrome c to the cytosol and activation of caspases

Materials and Methods: In this experimental study, we mainly investigated the effect of taraxerol on HeLa cells. We tested cell viability by the MTT assay and morphologic changes, analyzed apoptosis by DAPI staining and flow cytometry. We also determined reactive oxygen species [ROS] and mitochondrial membrane potential [MMP] using a Microplate Reader. In addition, the apoptotic proteins were tested by Western blot

Results: Taraxerol enhanced ROS levels and attenuated the MMP [delta psi m] in HeLa cells. Taraxerol induced apoptosis mainly via the mitochondrial pathway including the release of cytochrome c to the cytosol and activation of caspases 9 and 3, and anti-poly [ADP- ribose] polymerase [PARP]. Taraxerol could induce the down-regulation of the anti-apoptotic protein Bcl-2 and up-regulation of pro-apoptotic protein Bax. It suppressed the PI3K/ Akt signaling pathway

Conclusion: These results demonstrated that taraxerol induced cell apoptosis through a mitochondria-mediated pathway in HeLa cells. Thus, taraxerol might be a potential anticervical cancer candidate

Protein and mRNA expression of CTGF, CYR61, VEGF-C and VEGFR-2 in bone marrow of leukemia patients and its correlation with clinical features / 中国实验血液学杂志

This study was aimed to investigate the mRNA and protein expression of CTGF, CYR61, VEGF-C and VEGFR-2 in bone marrow of patients with leukemia, and to analyze the role and clinical significance of these 4 factors in genesis and development of leukemia, infiltration and metastasis of leukemic cells. A total of 100 cases of newly diagnosed leukemia, 26 cases of acute leukemia in complete remission and 30 controls were enrolled in this study. The mononuclear cells of bone marrow were collected, the mRNA and protein expression levels of CTGF, CYR61, VEGF-C, VEGFR-2 in leukemia patients and controls were detected by real time PCR and Western blot, respectively. The results showed that the mRNA and protein expression levels of above mentioned 4 factors were significantly higher than those in control (P < 0.05), only CTGF mRNA expression in AL patients after complete remission showed statistical difference as compared with control (P < 0.05), but the expression of CTGF mRNA showed statistical significance in different bone marrow hyperplasia of acute leukemia (P < 0.05). The expression level of CTGF protein showed difference in different chromosome karyotypes of leukemia (P < 0.05). The expression levels of CYR61 and VEGF-C proteins showed statistical difference in different bone marrow hyperplasia of acute leukemia (P < 0.05). The expression level of CTGF, CYR61, VEGF-C mRNA and protein in CML group were higher than that in control group. The expression levels of CTGF and CYR61 protein were higher than that in control. The mRNA and protein expression levels of above-mentioned 4 factors in sex and infiltration lf leukemic cells did not show statistical significance(P < 0.05). In correlative analysis, the mRNA expressions of above mentioned 4 factors were positively correlated with bone marrow blast count(P < 0.05), the protein expression of CTGF, CYR61 and VEGF-C were positively correlated with bone marrow blast count. It is concluded that the CTGF, CYR61, VEGF-C and VEGFR-2 mRNA and protein play a role in acute leukemia. In acute leukemia (AML/ALL), the expression of above mentioned factor was high, but except VEGFR-2. Most of them were positively correlated with bone marrow blast count. Joint block of these angiogenesis-related factors is likely to play an important role in targeting treatment of leukemia.

Study of Relationship of GM-CSF and Its Receptor with Early Spontaneous Abortion / 昆明医科大学学报

Objective To investigate the relationship of Maternal-fetal interface local cytokine GM-CSF and its receptor with the occurrence of early spontaneous abortion. Methods From August 2009 to December 2011,we collected 30 villi tissue samples with artificial abortion and 30 villi tissue samples with spontaneous abortion. At the same time, we collected 30 villi tissue samples with artificial abortion and 30 decidua tissue samples with spontaneous abortion,and 30 decidua tissue samples with spontaneous abortion. The human chorionic gonadotropin ( HCG) was detected by radioimmunoassay in every group. The expressions of granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte-macrophage colony-stimulating factor receptor (GM-CSFR ) were detected by Immunohistochemical staining and Western Blot in the villi tissue and decidua tissue in every group. Results The concentration of HCG in the spontaneous abortion group was lower than that in the artificial abortion group ( <0.05) . The protein expressions of GM-CSF and GM-CSFR were found in villus and deciduas tissues in both groups. The protein expression levels of GM-CSFR in the villus tissues were higher in spontaneous abortion group than those in artificial abortion group ( <0.05), the protein expression of GM-CSF was upregulated, but there was no statistically significant difference between two groups. In deciduas tissues, the protein expressions of GM-CSF and GM-CSFR were upregualted in spontaneous abortion group ( <0.05) . Conclusions The suitable concentrations of GM-CSF and GM-CSFR in decidua tissue maintain the pregnancy continued. However,the higher concentrations of GM-CSF and GM-CSFR in the decidua tissue may be one of reasons of spontaneous abortion.

Analysis of related factors influencing nurses'self-efficacy for palliative care / 中国实用护理杂志

Objective To explore the nurses'self-efficacy for palliative care and its related factors.Methods Data were collected via a self-constructed questionnaire,using the purposive sampling method.Subjects were nurses from two “grade-A” general hospitals in Henan province.Results The nurses' self-efficacy for palliative care stayed at a disequilibrium state,mean score of self-efficacy for physical care was 4.00,followed by family care self-efficacy 3.85,then psychological and spiritual care self-efficacy 3.70.Significant difference existed in self-efficacy for palliative care in nurses having different attitudes toward death.The cognitive level for palliative care,past experience of caring for end-stage patients were positively correlated with self-efficacy.Conclusions The main factors of the nurses' self-efficacy for palliative care related to attitudes toward death,past experience of caring for end-stage patients and the cognition level of nurses to palliative care.

Temporal lobe epilepsy with hypothalamic hamartoma: a rare case / 中华医学杂志(英文版)

Refractory gelastic seizure is one of the most common clinical manifestations in patients with hypothalamic hamartoma (HH) and HH is usually regarded as the epileptogenic focus. A young female patient with a small HH and refractory seizures is reported here. However, both the seizure semiology and results of electroencephalogram monitoring indicated the right temporal region was the epileptogenic focus. Thus a standard right anterior temporal lobectomy was performed while the hamartoma preserved. There was a marked improvement in both seizure frequency and quality of life during a 13-month follow-up. The outcome supported the concept that independent epileptogenic focus outside of the hypothalamus might occur in patients with HH.

Expression of heparanase and its coagulation proteins on the surface of leukemic cells / 白血病·淋巴瘤

Objective To explore whether the expression level of heparanase (HPA) and its coagulation proteins on leukemic blast membrane could determine the hemostatic balance on the surface of leukemia cells.Methods Forty patients of leukemia were studied,and 20 patients with iron dificient anemia as the control group.Expression of tissue factor (TF),heparanase (HPA),tissue factor pathway inhibitor (TFPI),and urokinase plasminogen activator receptor (UPAR) on leukemic blast surfaces were analyzed by flowcytometry.Results The expression of TF,UPAR,and HPA in AML,ALL,CML,CLL and CRAL groups were significantly higher compared with the control group (t =.3.289,3.507,2.701,P ＜0.05; t =2.498,0.802,3.090,P ＜0.05; t =2.642,3.308,2.696,P ＜0.05; t =3.417,3.434,2.382,P ＜0.05; t =2.193,2.272,2.263,P ＜0.05).There were no significantly differences between the leukemic cell expression of TFPI and the control group (P ＞0.05).Expression of TF,UPAR,HPA in AML patients were significantly higher than ALL,CML and CLL groups (t =2.463,2.179,2.276,P ＜0.05; t =2.637,2.402,2.095,P ＜0.05; t =2.548,2.425,2.412,P ＜0.05).The levels of TF,UPAR and HPA in M3,M4 and M5 patients were higher than that of M1,M2 groups (P ＜0.05).There were no significantly differences among M3,M4 and M5 (P ＞0.05).Conclusions These results suggest that TF,UPAR and HPA are predominately expressed on leukemic blast surface,particularly in M3and M4,5 subtypes.The expression of coagulation proteins on blast membrane might determine the hemostatic balance on the surface of leukemia cells.

Cut-off points of fasting fingertip capillary blood glucose for detecting both undiagnosed diabetes and pre-diabetes / 中华流行病学杂志

Objective To determine the efficient cut-off points of fasting fingertip blood glucose test for undiagnosed diabetes mellitus(DM), impaired glucose tolerance(IGT), and impaired fasting glucose(IFG)in community-based residents aged above 45 years old. Methods A cluster-randomized study was conducted from May 2008 to January 2009. A total of 3250 subjects aged above 45 years in two communities of Baoding city received questionnaire investigation and tested for fingertip blood glucose. Those subjects whose capillary blood glucose level ≥5.1 mmol/L were subjected to 75 g oral glucose tolerance test. Undiagnosed diabetes mellitus and pre- diabetes were identified by fasting plasma glucose and OGTT. In this study, the cut-off points of fasting capillary blood glucose for detecting undiagnosed diabetes and pre-diabetes were evaluated, using receiver operator characteristic curve(ROC). Results Of 1351 subjects that having had oral glucose tolerance test, 230 cases were diagnosed as diabetes mellitus(7.3%), 166 cases(5.2%)as IFG, and 204(6.7%)as IGT under fasting capillary blood glucose as test variable and state variables according to the following criteria.(1)FPG≥7.0 mmol/L or/and 2hPG≥11.1 mmol/L(2)FPG<5.6 mmol/L (3)FPG<7.0 mmol/L and 7.8 mmol/L≤2hPG≤ 11.1 mmol/L, areas under three ROC curves were 0.905, 0.633 and 0.719, respectively. The cut-offvalues of screening for undiagnosed DM, IGT and IFG were 6.0 mmol/L, 5.7 mmol/L, and 5.7 mmol/L, respectively. When cut-off value of screening for undiagnosed DM was 6.0 mmol/L, the maximal sensitivity was 78.0% and specificity was 89.3%.But there were both lower sensitivity and specificity in screening for IFG and IGT according to the best predicting value(5.7 mmol/L)from the ROC curves(50.3% and 28.0% vs. 60.8% and 28.0%). Conclusion Fasting capillary blood glucose with the lower cut-point of 6.0 mmol/L in screening for undiagnosed diabetes mellitus alone, was relatively reliable, whereas for both IFG and IGT the fasting fingertip blood glucose tests were fallible. It was convenient and could be used in screening the DM at the community level.

Expression of human annexin V in different fetal tissues / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of human annexin-V (HA-V) in relation to HBV infection in different fetal tissues.</p><p><b>METHODS</b>Immunohistochemistry was employed to detect the expression and distribution of HA-V in the liver, kidney, ovary, heart, fallopian tube, spleen, and thymus gland of human fetus.</p><p><b>RESULTS</b>HA-V expression was detected in different tissues including the ovary, liver, intrahepatic bile duct, heart, kidney, lymphocytic cells in the thymus gland, epithelial cells of the fallopian, and cortical and medullary cells of the spleen. HA-V was distributed mainly in the cytoplasm of the cells. The liver tissues exhibited greater gray scale for HA-V expression than in the other tissues (P<0.05) and no significant difference was observed in the other tissues than the liver (P>0.05) in image analysis with Photoshop 7.0.</p><p><b>CONCLUSION</b>HA-V is an inherent protein in fetal tissues with possible relation to HBV infection of different tissues as a HBV receptor. Greater amount of HA-V in the liver may account for the vulnerability of the liver to HBV infection.</p>

Clinical significance of detecting neonatal peripheral blood mononuclear cells infected by HBV / 中华儿科杂志

<p><b>OBJECTIVE</b>To understand the HBV infection rate of peripheral blood mononuclear cells (PBMCs) from fetuses of HBsAg positive mothers, associated risk factors and to explore the clinical significance of detecting HBV infected PBMCs.</p><p><b>METHODS</b>Sixty eight pregnant women who were delivered at the First Hospital of Xi'an Jiaotong University, China from August 1995 to February 1997, and their newborns were studied. They were divided into two groups according to their status of HBV serological markers. The study group included 50 cases who were serum HBsAg positive and 18 cases without any HBV serum markers served as control group. All these cases had no symptoms of hepatitis, high risk premature labor, premature delivery and hypertensive disorder complicating pregnancy. Age and gestational age were matched in two groups. Blood samples (5 mL) were taken from the peripheral vein of pregnant women before delivery and from newborns within 24 h after birth, before inoculation of HBV vaccine (HBVac) and injection of hepatitis B immunoglobulin (HBIG). PBMCs were isolated. The sera and PBMCs were stored at -80 degrees C. HBV-DNA in serum and PBMCs were detected with nested polymerase chain reaction (n-PCR). Two pairs of oligonucleotide primers, the outer primer pair for first PCR and inner primer pair for second PCR, designed according to region S of HBV genome were synthesized by Shanghai Cell Biology Institute of Chinese Academy of Science.</p><p><b>RESULTS</b>The detection rate of HBV-DNA in serum and PBMCs from HBsAg positive pregnant women was 60.0% (30/50) and 40.0% (20/50), respectively. The detection rate of HBV-DNA in serum and PBMCs from newborns of HBsAg positive pregnant women was 46.0% (23/50) and 30.0% (15/50), respectively. Ten newborns were HBV-DNA positive in serum only, 2 were positive in PBMCs only and 13 were positive in both serum and PBMCs. In the control group, HBV-DNA was not detected in PBMC nor in serum. The positive rate of HBV-DNA in PBMCs of newborns was significantly higher in the group of mothers who were HBV-DNA or HBeAg positive in serum (P < 0.05, P < 0.01); the positive rate was significantly higher in the group of mothers who were HBV-DNA positive in both serum and PBMC than that in the group of mothers who were serum HBV-DNA positive only (P < 0.01); and it was markedly higher in the group of mothers who were PBMC HBV-DNA positive than that in group of mothers who were HBV-DNA negative in PBMCs (P < 0.01). The positive rate of HBV-DNA in PBMCs of newborns was significantly higher in the group of newborns who were HBV-DNA positive in serum than that in the group of newborns who were HBV-DNA negative in serum (P < 0.05).</p><p><b>CONCLUSIONS</b>The positive rate of HBV-DNA in PBMCs from newborns of HBsAg positive pregnant women was 30.0% (15/30). It was related to HBV viremia level and HBV-DNA status in PBMCs of mothers and newborns. Detection of HBV-DNA in PBMCs may be an important supplementary method to determine intrauterine HBV infection, and can predict the response to HBV vaccine.</p>

Screening and cloning of hepatitis C virus non-structural protein 4B interacting protein gene in hepatocytes / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate biological functions of non-structural protein 4B (NS4B) of hepatitis C virus (HCV), yeast-two hybrid technique was performed to seek proteins in hepatocytes interacting with HCV NS4B.</p><p><b>METHODS</b>HCV NS4B bait plasmid was constructed by ligating the NS4B gene with carrier plasmid pGBKT7 and transformed into yeast cells AH109 (type alpha). The transformed yeast cells were amplified and mated with yeast cells Y187 (alpha type) containing liver cDNA library plasmid pACT2 in 2 x YPDA medium. Diploid yeast cells were plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) and synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-alpha-gal for selecting two times. After extracting plasmid from blue colonies, plasmid DNA was transformed into competent Escherichia coli and analysed by DNA sequencing and bioinformatics.</p><p><b>RESULTS</b>Five genes in eight positive colonies were obtained. There were one NADH dehydrogenase subunit 3, one cytochrome c oxidase subunit III, one retinol binding protein 4, one reticulon 3-A (RTN3) and one fibrinogen gamma polypeptide (FGG).</p><p><b>CONCLUSION</b>Genes of HCV NS4B interacting proteins in hepatocytes were successfully cloned and the results paved the way for studying the biological functions of NS4B and associated proteins.</p>

Screening and cloning of hepatitis C virus non-structural protein 4A interacting protein gene in hepatocytes / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate biological functions of hepatitis C virus (HCV) non-structural protein 4A (NS4A).</p><p><b>METHODS</b>Yeast-two hybrid technique was performed to seek proteins in hepatocytes interacting with HCV NS4A. HCV NS4A bait plasmid was constructed by ligating the NS4A gene with carrier plasmid pGBKT7, then it was transformed into yeast AH109 (alpha type). The transformed yeast cells were amplified and mated with yeast cells Y187 (alpha type) containing liver cDNA library plasmid pACT2 in 2 x YPDA medium. Diploid yeast cells were plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) and synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing X-alpha-gal for selection two times. After extracting plasmid from blue colonies, plasmid DNA was transformed into competent E.coli and analyzed by DNA sequencing and bioinformatics methods.</p><p><b>RESULTS</b>Among twenty-two positive colonies there were eleven positive for metallothionein 2A, three for eukaryotic translation elongation factor 1 alpha 1, two for albumin, two for RNA binding motif protein 21, two for myomesin, one for cytochrome C oxidase II, and one for ATPase.</p><p><b>CONCLUSIONS</b>Genes of HCV NS4A interacting proteins in hepatocytes were successfully cloned and the results pave the way for studying the biological functions of NS4A and associated proteins.</p>

FDG-positron emission tomography and EEG:The comparison in localizing in refractory epilepsy / 中国康复理论与实践

@#ObjectiveTo study the values of EEG and [18F]2-deoxyglucose(FDG) positron emission tomography in localizing the epiletogenic cortex,and evaluate their relation.MethodsVideo-EEG(VEEG) and FDG-PET scans were performed in 44 patients with refractory epilepsy.Electrocorticography(ECoG) in surgery and patholopy were performed in 38 patients who had undergone neurosurgical therapy.Congruence among them were studied.Results43 patients(98%) had FDG-PET hypometabolism.42 patients(95%) had epileptiform wave.There were 22 patients(50%) whose PET and EEG were in complete congruence,whereas 10 patients(23%) in part congruence.12 patients who had undergone operation had controversial results in PET and EEG,8 cases had agreement between ECoG and PET,and 2 cases had agreement between ECoG and VEEG. ConclusionFDG-PET is a effective,sensitive and non-invasive investigation.It can provide valuable supplemental data in patients with unlocalized surface EEG.

Clinical analysis of retinoic acid syndrome developed in 11 patients with acute promyelocytic leukemia / 中国实验血液学杂志

To explore the clinical features, risk factors an d treatment of retinoic acid syndrome (RAS) in patients with acute promyelocytic leukemia (APL) treated with retinoic acid, the clinical and laboratory data of 11 APL patients with RAS were retrospectively analysed. The results showed that earlier and more common symptoms of RAS were successively dyspnea (11/11), fever (10/11) and hydrothorax (6/11). Higher WBC count (> or = 15.0 x 10(9)/L) in the course of treatment of all-trans retinoic acid susceptible to develop RAS (9/11). The RAS patients were treated with dexamethasone without discontinuing the treatment of retinoic acid, complete remission was achieved in 10 cases and one patient died from disseminated intravascular coagulation. It is concluded that the identification and dexamethasone treatment of RAS in earlier period are extremely important for obtaining better clinical curative effect, and it does not influence therapeutic effect of continuing application of retinoic acid.

Screening of differentially expressed genes in placentas with hepatitis B virus infection by suppression subtractive hybridization technique / 中华妇产科杂志

Objective To screen differentially expressed genes in placentas with hepatitis B virus (HBV)infection and to discuss the molecular mechanism of HBV intrauterine infection.Methods Thirty placenta tissue specimens from HBsAg and HBV DNA positive pregnant women were used as the study group and 30 placenta tissue specimens from normal pregnant women with HBsAg and HBV DNA negativity were served as the control group.The suppression subtractive hybridization(SSH)technique was used.Total RNAs of placenta tissue of the study group were mixed as the tester,and total RNAs of placenta tissue of the control group were mixed as the driver.A subtractive cDNA library was constructed by PCR-selective cDNA subtraction systems.Amplifications of the library were carried out with E.coil strain DH5? by reverse spot hybridization.RT-PCR confirmed that phosphatidylinositol 3-kinase(PI3K)was up-regulated in placenta tissue with HBV infection.Results Colony PCR showed that the clones contained 200-1000 bp inserts. Thirty five clones were confirmed by reverse spot hybridization and analyzed by sequencing and bioinformatics.Thirty three known genes and 2 genes with unknown function were obtained.RT-PCR preliminarily confirmed that PI3K gene was up-regulated in HBV infected placenta.Conclusions The differentially expressed genes in placentas with hepatitis B virus(HBV)infection using SSH technique has been screened out successfully.These differentially expressed genes encoding proteins participating in cell vital metabolism and malformation,and signal conduction-antiapoptosis pathway.This finding brings some new clues for studying the mechanisms of HBV intrauterine infection.