[Research advancement of long non-coding RNAs in liver fibrosis].

Hepatic fibrosis is a wound healing and scar repair reaction after liver injury, and is a common pathway for various chronic liver diseases. Activation and proliferation of hepatic stellate cells are the key links in the occurrence and development of hepatic fibrosis. Recent studies have shown that long non-coding RNAs are involved in regulating the activation, proliferation and apoptosis of hepatic stellate cells. Thus, probing its mechanism of action will provide a new strategy for the diagnosis, treatment and prognosis in liver fibrosis.

Diagnostic value of joint detection of homocysteine and RDW CV on acute myocardial infarction.

The article "Diagnostic value of joint detection of homocysteine and RDW CV on acute myocardial infarction" by G.-X. Hu, J. Zhang, Y.-G. Tian, Y.-H. Li, L. Mou, L.-J. Qiao, published in Eur Rev Med Pharmacol Sci 2016; 20 (19): 4124-4128 has been withdrawn.

Diagnostic value of joint detection of homocysteine and RDW CV on acute myocardial infarction. Retracted.

OBJECTIVE: We discussed the diagnostic value of joint detection of homocysteine (HCY) and red blood cell volume distribution width variable coefficient on acute myocardial infarction (AMI). PATIENTS AND METHODS: We collected 300 coronary heart disease cases, among which there were 121 cases of stenocardia, 65 cases of ischemic heart failure, and 114 cases of AMI at the Department of Cardiology of our hospital during the period from January 2012 to June 2013. At the same time, we took 100 normal physical examinees as the control group, used the full-automatic cell-analyzer and the immunization to measure HCY and red blood cell distribution width (RDW) CV respectively and analyze their value in diagnosing AMI. RESULTS: The differences among the four groups of HCY and RDW CV were statistically significant (p < 0.05). The HCY and RDW CV level in the AMI group were significantly higher than those of the other three groups (p < 0.05); the differences between the positive diagnosis rate of HCY, the RDW CV and their joint diagnosis in the AMI group were statistically significant (p < 0.05) while the differences between the positive diagnosis rate of HCY, the RDW CV and their joint diagnosis in the control group were not statistically significant (p > 0.05). The detection sensitivity and specificity of HCY alone were respectively 68.42% and 86.00% with those of the RDW CV alone being 64.91% and 84.00%. The joint detection sensitivity and specificity were 83.33% and 93.00%, statistically different (p < 0.05). The concordance rate, the positive predictive value and the negative predictive value were 87.85%, 93.14% and 83.04%, respectively. CONCLUSIONS: The HCY and RDW CV joint diagnosis of AMI had relatively high sensitivity, specificity, concordance rate, positive predictive value and negative predictive value.

Bovine Mx1 enables resistance against foot-and-mouth disease virus in naturally susceptible cells by inhibiting the replication of viral RNA.

Innate immunity, especially the anti-viral genes, exerts an important barrier function in preventing viral infections. Myxovirus-resistant (Mx) gene take an anti-viral role, whereas its effects on foot-and-mouth disease virus (FMDV) in naturally susceptible cells are still unclear. The bovine primary fetal tracheal epithelial cell line BPTE-siMx1, in which bovine Mx1 gene was silenced, was established and treated with IFN alpha for 6 hr before FMDV infection. The copy numbers of the negative and positive strand viral RNA were determined by strand-specific real-time fluorescence quantitative RT-PCR. The TCID50 of BPTE-siMx1 cells increased at least 17-fold as compared to control cells BPTE-LacZ at 8 hr post infection, thus silencing of bovine Mx1 could promote the replication of FMDV. The amount of both the negative and positive strand viral RNA in BPTE-siMx1 cells significantly increased as compared to BPTE-LacZ cells, indicating that the replication levels of viral RNA were promoted by silencing bovine Mx1. The bovine Mx1 gene could provide resistance against FMDV in the bovine primary fetal tracheal epithelial cells via suppressing the replication of viral RNA.

Protective effects against and potential mechanisms underlying the effect of magnesium isoglycyrrhizinate in hypoxia-reoxygenation injury in rat liver cells.

We examined the protective effects of magnesium isoglycyrrhizinate (MgIG) on hypoxia-reoxygenation injury in rat liver cells. Rat liver cells in the logarithmic growth phase were divided into the hypoxia-reoxygenation injury model group and MgIG pretreatment group (0.01, 0.1, 1, 10, 100 mg/mL). After 24-h pretreatment, we detected the effects of MgIG on liver cell viability using the methyl thiazolyl tetrazolium (MTT) assay at 6-h hypoxia and 4-h reoxygenation. After 24-h pretreatment, liver cells were randomly divided into the hypoxia-reoxygenation injury model group and low-, moderate-, and high-MgIG-concentration groups (0.1, 1, 10 mg/mL, respectively), and hypoxia and reoxygenation were simulated for 6 and 4 h, respectively. Cell morphology was observed by light microscopy. Nuclear factor-kB gene expression was analyzed by quantitative reverse transcription-polymerase chain reaction. MTT results showed that MgIG (0.1, 1, 10 mg/mL) improved the A-value of anoxia-reoxygenation injury in liver cells (P < 0.01) compared with that of the model group. Cells did not survive when the MgIG concentration was 100 mg/mL. At an MgIG concentration lower than 0.01 mg/mL, the A-value of the MTT group was higher than that of the model group (P > 0.05). Nuclear factor-kB mRNA expression (0.597 ± 0.062, 0.248 ± 0.067, 0.141 ± 0.029) in the low-, moderate-, and high-concentration groups was lower than that in the model group (P < 0.01). MgIG reduced hypoxia-reoxygenation injury of liver cells, indicating that it improved hepatic cell activity, inhibited lipid peroxidation and inflammatory reactions, and decreased nuclear factor-kB mRNA expression.

Effects of hydroxycamptothecin on the expression of matrix metalloproteinase-1 (MMP-1), tissue inhibitor of MMP-1, and type I collagen in rats with pulmonary fibrosis.

The aim of this study was to investigate the effects of hydroxycamptothecin (HCPT) on the expression of matrix metalloproteinase-1 (MMP-1), tissue inhibitor of MMP-1 (TIMP-1), and type I collagen in the lung tissue of rats with pulmonary fibrosis induced by bleomycin A5. We used hematoxylin eosin staining to observe the degree of pulmonary fibrosis in rats; Masson staining, reverse transcription polymerase chain reaction, and immunohistochemistry were used to observe the expression of collagen, MMP-1, and TIMP-1, and type I collagen. The expression of MMP-1 in the model group decreased significantly, while the expression of TIMP-1 and type I collagen significantly increased. After treatment with HCPT, the degree of pulmonary fibrosis and the expression of TIMP-1 and type I collagen decreased in all treatment groups. However, the expression of MMP-1 increased in a dose-dependent manner. Our results showed that HCPT decreased the pulmonary fibrosis induced by bleomycin A5 in rats, and an increase in MMP-1 expression and decrease in the TIMP-1 and type I collagen expression may be the mechanism that regulates the metabolism of the extracellular matrix.

293FT is a highly suitable mammalian cell line for the in vitro enzymatic activity analysis of typical P450 proteins.

Mammalian cells have been widely used for the in vitro evaluation of the functional effect of allelic variants of cytochrome P450 (CYP). The aim of this study was to determine the most suitable mammalian cell line for the in vitro drug metabolism analysis of CYP variants. Three reported cell lines (COS-7, HepG2, 293T) and one fast-growing variant of the 293 cell line 293FT were transfected with vectors expressing green fluorescent protein or typical variants of CYP2C9, CYP2C19 or CYP2D6 to investigate the protein expression levels and the catalytic activity of expressed CYP allelic variants. The transfected 293FT cells had the highest protein expression level and exhibited the highest enzymatic activity, while HepG2 cells showed the lowest activity among the four tested cell lines. Simultaneously, 293FT cells still maintained the similar relative enzymatic ratio among three typical CYP2C9 variants to that of the commonly used COS-7 cells. In addition, 293FT cells could also be used for the in vitro functional evaluation of two other typical P450 proteins, CYP2C19 and CYP2D6. Therefore, the 293FT cell line is more suitable for the in vitro enzymatic activity analysis of typical P450 proteins than any other reported mammalian cell lines.

CYP2C9 polymorphism analysis in Han Chinese populations: building the largest allele frequency database.

Genetic polymorphisms of CYP2C9 significantly influence the pharmacokinetics and pharmacodynamics of some drugs, which might result in adverse drug effects and therapeutic failure. Several studies have been performed on CYP2C9 genetic polymorphisms in Han Chinese populations. However, these studies only focused on two commonly investigated alleles, *2 and *3, in relatively small sample sizes. To scale up the gene-scanning region and determine relatively precise data on the genetic distribution pattern in Chinese populations, unrelated healthy Han Chinese volunteers from Zhejiang Province (n=1127) and Hebei (n=1000) Province were recruited as subjects for the direct sequencing of all exons of CYP2C9. As a result, 14 previously reported alleles were detected in this work, and 8 of these alleles (*14, *16, *19, *23, *27, *29, *33 and *34) were described for the first time in Chinese populations. In addition, 37 novel mutations were also detected, of which 22 variants were non-synonymous, and 21 new alleles, *36-*56, were designated by the Human CYP Allele Nomenclature Committee. In vitro functional analysis of these 22 novel CYP2C9 variants revealed that 17 mutations had a significant influence on the protein's catalytic activity. Our study provides the most accurate data on CYP2C9 polymorphisms in Han Chinese populations and detects the largest number of novel allelic variants existing to date. These new alleles will greatly enrich the current knowledge of naturally occurring CYP2C9 variants in Chinese populations.

Effects of methoxychlor and 2,2-bis(p-hydroxyphenyl)-1,1,1-trichloroethane on 3ß-hydroxysteroid dehydrogenase and 17ß-hydroxysteroid dehydrogenase-3 activities in human and rat testes.

Human and rat testis microsomes were used to investigate direct inhibitory activities of methoxychlor (MXC) and its metabolite 2,2-bis(p-hydroxyphenyl)-1,1,1-trichloroethane (HPTE) on 3ß-hydroxysteroid dehydrogenase (3ß-HSD) and 17ß-hydroxysteroid dehydrogenase type 3 (17ß-HSD3). The 3ß-HSD and 17ß-HSD3 enzymes are involved in the reactions that culminate in androgen biosynthesis in Leydig cells. The results demonstrated that MXC and HPTE inhibited human 3ß-HSD activity at a concentration of 10 nm. The half maximal inhibitory concentration (IC(50) ) for MXC inhibition of 3ß-HSD was 53.21 ± 15.52 µm (human) and 46.15 ± 17.94 µm (rat), and for HPTE, it was 8.29 ± 2.49 µm (human) and 13.82 ± 2.26 µm (rat). At the higher concentration of 100 µm, MXC did not affect human and rat 17ß-HSD3 activity. However, the IC(50) for HPTE inhibition of 17ß-HSD3 was 12.1 ± 1.9 µm (human) and 32 .0 ± 8.6 µm (rat). The mode of action of MXC and HPTE on 3ß-HSD activity was non-competitive with the substrate pregnenolone, but was competitive with the cofactor NAD(+) . The mode of HPTE inhibition of 17ß-HSD3 was non-competitive with the substrate androstenedione, but was competitive with the cofactor NADPH. In summary, our results showed that HPTE, which is the biologically active metabolite of MXC, has the capacity for direct inhibition of 3ß-HSD and 17ß-HSD3 enzyme activity. Inhibition of enzyme activity is presumably associated with suppression of steroidogenesis in gonadal tissues and has implications for testis function.

Effect of sodium dimercaptopropanesulfonate on antagonism of tetramethylenedisulphotetramine to GABA receptor.

AIM: To study effects of sodium dimercaptopropanesulfonate (DMPS) on the antagonism of tetramethylenedisulphotetramine (TETS) to gamma-aminobutyric acid (GABA) receptor. METHODS: Acute toxicity experiments were conducted to observe the effects of DMPS and TETS on mice. Contents of free amino acids in mouse brain were determined with automatic analyzer for amino acids. Autoradiography was used to observe the [3H]GABA bindings in the rat brain slices under different conditions. RESULTS: After icv and ip DMPS, the number of mice experiencing convulsions reduced from 20 in control group to 4 and 2 respectively in TETS poisoned mice. The content of GABA was altered in DMPS control group and TETS control group compared with DMPS protection group and NS control group [micromol/g: (2.09 +/- 0.05) and (2.67 +/- 0.15) vs (2.40 +/- 0.10 (micromol/g)) and (2.41 +/- 0.21)]; the content of glutamic acid was (12.3 +/- 1.2), (12.0 +/- 0.8), (10.2 +/- 0.6), and (11.8 +/- 1.0) micromol/g in NS control group, DMPS control group, TETS control group, and DMPS protection group, respectively. The OD value of autoradiograms decreased in TETS group compared with buffer control group in cortex, hippocampus, diencephalon, and brainstem [(0.084 +/- 0.008), (0.081 +/- 0.009), (0.094 +/- 0.006) and (0.081 +/- 0.006), vs (0.102 +/- 0.003), (0.109 +/- 0.005), (0.128 +/- 0.007), and (0.125 +/- 0.008), respectively]. OD value was maintained or higher than the normal level in DMPS+TETS group in the four brain areas [(0.116 +/- 0.008), (0.125 +/- 0.011), (0.129 +/- 0.005), and (0.128 +/- 0.010) vs (0.102 +/- 0.003), (0.109 +/- 0.005), (0.128 +/- 0.007), and (0.125 +/- 0.008), respectively]. CONCLUSION: The inhibitory effects of DMPS on the antagonism of TETS to GABA receptor are due to the increase in the GABA binding to its receptors in brain caused by DMPS

Insight into hepatocellular carcinogenesis at transcriptome level by comparing gene expression profiles of hepatocellular carcinoma with those of corresponding noncancerous liver.

Human hepatocellular carcinoma (HCC) is one of the most common cancers worldwide. In this work, we report on a comprehensive characterization of gene expression profiles of hepatitis B virus-positive HCC through the generation of a large set of 5'-read expressed sequence tag (EST) clusters (11,065 in total) from HCC and noncancerous liver samples, which then were applied to a cDNA microarray system containing 12,393 genes/ESTs and to comparison with a public database. The commercial cDNA microarray, which contains 1,176 known genes related to oncogenesis, was used also for profiling gene expression. Integrated data from the above approaches identified 2,253 genes/ESTs as candidates with differential expression. A number of genes related to oncogenesis and hepatic function/differentiation were selected for further semiquantitative reverse transcriptase-PCR analysis in 29 paired HCC/noncancerous liver samples. Many genes involved in cell cycle regulation such as cyclins, cyclin-dependent kinases, and cell cycle negative regulators were deregulated in most patients with HCC. Aberrant expression of the Wnt-beta-catenin pathway and enzymes for DNA replication also could contribute to the pathogenesis of HCC. The alteration of transcription levels was noted in a large number of genes implicated in metabolism, whereas a profile change of others might represent a status of dedifferentiation of the malignant hepatocytes, both considered as potential markers of diagnostic value. Notably, the altered transcriptome profiles in HCC could be correlated to a number of chromosome regions with amplification or loss of heterozygosity, providing one of the underlying causes of the transcription anomaly of HCC.

Cloning and functional analysis of cDNAs with open reading frames for 300 previously undefined genes expressed in CD34+ hematopoietic stem/progenitor cells.

Three hundred cDNAs containing putatively entire open reading frames (ORFs) for previously undefined genes were obtained from CD34+ hematopoietic stem/progenitor cells (HSPCs), based on EST cataloging, clone sequencing, in silico cloning, and rapid amplification of cDNA ends (RACE). The cDNA sizes ranged from 360 to 3496 bp and their ORFs coded for peptides of 58-752 amino acids. Public database search indicated that 225 cDNAs exhibited sequence similarities to genes identified across a variety of species. Homology analysis led to the recognition of 50 basic structural motifs/domains among these cDNAs. Genomic exon-intron organization could be established in 243 genes by integration of cDNA data with genome sequence information. Interestingly, a new gene named as HSPC070 on 3p was found to share a sequence of 105bp in 3' UTR with RAF gene in reversed transcription orientation. Chromosomal localizations were obtained using electronic mapping for 192 genes and with radiation hybrid (RH) for 38 genes. Macroarray technique was applied to screen the gene expression patterns in five hematopoietic cell lines (NB4, HL60, U937, K562, and Jurkat) and a number of genes with differential expression were found. The resource work has provided a wide range of information useful not only for expression genomics and annotation of genomic DNA sequence, but also for further research on the function of genes involved in hematopoietic development and differentiation.

Gene expression networks underlying retinoic acid-induced differentiation of acute promyelocytic leukemia cells.

To elucidate the molecular mechanism of all-trans-retinoic acid (ATRA)-induced differentiation of acute promyelocytic leukemia (APL) cells, the gene expression patterns in the APL cell line NB(4) before and after ATRA treatment were analyzed using complementary DNA array, suppression-subtractive hybridization, and differential-display-polymerase chain reaction. A total of 169 genes, including 8 novel ones, were modulated by ATRA. The ATRA-induced gene expression profiles were in high accord with the differentiation and proliferation status of the NB(4) cells. The time courses of their modulation were interesting. Among the 100 up-regulated genes, the induction of expression occurred most frequently 12-48 hours after ATRA treatment, while 59 of 69 down-regulated genes found their expression suppressed within 8 hours. The transcriptional regulation of 8 induced and 24 repressed genes was not blocked by cycloheximide, which suggests that these genes may be direct targets of the ATRA signaling pathway. A balanced functional network seemed to emerge, and it formed the foundation of decreased cellular proliferation, maintenance of cell viability, increased protein modulation, and promotion of granulocytic maturation. Several cytosolic signaling pathways, including JAKs/STAT and MAPK, may also be implicated in the symphony of differentiation. (Blood. 2000;96:1496-1504)

Nuclear pseudogenes of mitochondrial DNA as a variable part of the human genome.

Novel pseudogenes homologous to the mitochondrial (mt) 16S rRNA gene were detected via different approaches. Eight pseudogenes were sequenced. Copy number polymorphism of the mtDNA pseudogenes was observed among randomly chosen individuals, and even among siblings. A mtDNA pseudogene in the Y-chromosome was observed in a YAC clone carrying only repetitive sequence tag site (STS). PCR screening of human yeast artificial chromosome (YAC) libraries showed that there were at least 5.7 x 10(5) bp of the mtDNA pseudogenes in each haploid nuclear genome. Possible involvement of the mtDNA pseudogenes in the variable part of the human nuclear genome is discussed.

[Antidotal effects of 2,3-dimercaptopropane-1-sulfonate sodium (DMPS) and combined with diazepam on acute poisoning caused by sodium ammonium dimethyl-2-propano-1,3-dithiosulfate monohydrate (SCD)].

In mice, DMPS (250 mg/kg, i.v.) combined with diazepam (1.25 mg/kg, i.p.) could increase LD50 of p. o. SCD 5.3 times. DMPS (62.5 mg/kg, i.v.) antagonized completely the respiratory depression and neuromuscular blockade caused by SCD(7.5 mg/kg, i.v.) in rabbits. SCD (15 mg/kg, i.v.) caused tremor, tonic convulsion and the abnormal paroxysmal discharges in EEG in rabbits. DMPS (0.5 mg/kg, i.c.v) could not eliminate the abnormal paroxysmal discharges in EEG of rabbits. DMPS (62.5 mg/kg, i.v.) combined with diazepam (5 mg/kg, i.v.) completely and rapidly antagonize these toxic symptoms and the abnormal changes in EEG.

Studies on the calcium antagonistic action of tetrandrine: X VI. Hemodynamic effects of tetrandrine on conscious rats.

Hemodynamic study of tetrandrine (Tet) in conscious rats showed that 15 mg/kg i.v. lowered BP, LVSP, +/- dp/dtmax, and (dP/dt)P-1. The degree of diminutions was nearly equal to that of BP during the initial period, while the LVEDP was elevated. But all these parameters (except -dp/dtmax) recovered gradually and earlier than BP, and LVEDP decreased to a level slightly lower than that of control. These results indicate that the hypotensive action of Tet is mainly due to its inhibition of cardiac contractility at the early period, but due to vasodilatation in the later stage. The HR was slowed down abruptly followed by a reflex acceleration, and than a gradual but sustained decrease in HR supervened with i.v. Tet. When large dose (40 mg/kg) of Tet i.v. caused a cardiac standstill with the R wave of ECG persisting for a few minutes, it means that an excitation-contraction decoupling occurred as that found on isolated myocardial preparation treated with verapamil and Tet.