MiR-155-5p affects Wilms' tumor cell proliferation and apoptosis via targeting CREB1.

The article "MiR-155-5p affects Wilms' tumor cell proliferation and apoptosis via targeting CREB1", by X.-S. Zhao, B. Han, J.-X. Zhao, N. Tao, C.-Y. Dong, published in Eur Rev Med Pharmacol Sci 2019; 23 (3): 1030-1037-DOI: 10.26355/eurrev_201902_16990-PMID: 30779069, has been retracted by the authors due to a slight deviation in the data. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/16990.

Long noncoding RNA MIAT acts as an oncogene in Wilms' tumor through regulation of DGCR8.

The article "Long noncoding RNA MIAT acts as an oncogene in Wilms' tumor through regulation of DGCR8, by X.-S. Zhao, N. Tao, C. Zhang, C.-M. Gong, C.-Y. Dong, published in Eur Rev Med Pharmacol Sci 2019; 23 (23): 10257-10263-DOI: 10.26355/eurrev_201912_19663-PMID: 31841180" has been withdrawn from the authors due to inaccuracies in the data. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/19663#:~:text=CONCLUSIONS%3A%20The%20above%20results%20suggested,and%20therapy%20of%20Wilms'%20tumor.

Long noncoding RNA SNHG16 acts as an oncogene in Wilms' tumor through sponging miR-200a-3p.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "Long noncoding RNA SNHG16 acts as an oncogene in Wilms' tumor through sponging miR-200a-3p, by X.-S. Zhao, N. Tao, C. Zhang, C.-M. Gong, C.-Y. Dong, published in Eur Rev Med Pharmacol Sci 2020; 24 (8): 4145-4151-DOI: 10.26355/eurrev_202004_20994-PMID: 32373950" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/20994.

Long noncoding RNA SNHG16 acts as an oncogene in Wilms' tumor through sponging miR-200a-3p.

OBJECTIVE: Recently, the role of long noncoding RNA (lncRNAs) in tumor progression has attracted much attention. The aim of this study was to investigate the role of lncRNA SNHG16 in the development of Wilms' tumor, and to explore the underlying mechanism. PATIENTS AND METHODS: Real Time-quantitative Polymerase Chain Reaction (RT-qPCR) was used to detect SNHG16 expression in Wilms' tumor patients' tissues. Function assays, including wound healing assay, and transwell assay, were conducted to detect the changes of biological behaviors in Wilms' tumor cells due to gain or loss of SNHG16. Besides, the luciferase reporter gene assay was performed to explore the underlying mechanism. RESULTS: The expression level of SNHG16 was significantly up-regulated in Wilms' tumor tissues when compared with adjacent tissues. Cell migration and invasion abilities were significantly repressed via down-regulation of SNHG16. However, opposite results were obtained after up-regulation of SNHG16 in vitro. After the down-regulation of SNHG16, the expression of miR-200a-3p increased significantly. However, the expression of miR-200a-3p was remarkably reduced via up-regulation of SNHG16 in vitro. Furthermore, SNHG16 acted as a competing endogenous RNA via sponging miR-200a-3p in Wilms' tumor. CONCLUSIONS: SNHG16 induced the metastasis of Wilms' tumor via sponging miR-200a-3p. Our findings might provide a new prospect for the diagnosis and therapy of Wilms' tumor.

Long noncoding RNA MIAT acts as an oncogene in Wilms' tumor through regulation of DGCR8.

OBJECTIVE: Recent researches have proved that long noncoding RNAs (lncRNAs) cover an important role in malignant tumors. Our study showed how lncRNA myocardial infarction-associated transcript (MIAT) functions in the development of Wilms' tumor. PATIENTS AND METHODS: Real Time-quantitative Polymerase Chain Reaction (RT-qPCR) was utilized to detect the MIAT expression in Wilms' tumor patients. The MIAT expression level and the patients' overall survival time were analyzed. Then, we conducted functional experiments to identify the changes in the biological behaviors of Wilms' tumor cells due to the loss of MIAT. Moreover, further experiments were performed to explore the potential mechanism. RESULTS: By comparing with MIAT expression in adjacent tissues, the MIAT expression level was significantly higher in Wilms' tumor samples. Moreover, the cell growth ability of Wilms' tumor cells was inhibited due to the loss of MIAT. The migrated and invaded ability of the Wilms' tumor cells was inhibited due to the loss of MIAT. Furthermore, the expression of DGCR8 was downregulated due to the loss of MIAT. In addition, it was found that the DGCR8 expression was positively correlated to MIAT expression in Wilms' tumor tissues. CONCLUSIONS: The above results suggested that MIAT could promote the cell proliferation and the metastasis of Wilms' tumor by upregulating DGCR8, which indicated that MIAT might be a potential target for the diagnosis and therapy of Wilms' tumor.

MiR-155-5p affects Wilms' tumor cell proliferation and apoptosis via targeting CREB1.

OBJECTIVE: The aim of this study was to explore the role of microRNA-155-5p (miR-155-5p) in regulating the proliferation and apoptosis of Wilm's tumor (WT), and to investigate the possible underlying mechanism. PATIENTS AND METHODS: The expression levels of miR-155-5p in 37 pairs of WT clinical samples, as well as WT cell line (G401), were detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay and flow cytometry assay were used to detect the effects of miR-155-5p on cell proliferation, cycle and apoptosis. Target gene prediction software was applied to screen the potential downstream target gene of miR-155-5p. QRT-PCR, Western blot (WB) and luciferase reporter gene assay proved that cAMP-response element binding protein 1 (CREB1) was the target gene of miR-155-5p. Besides, rescue experiment was conducted to further explore the effect of CREB1 on WT cells. RESULTS: The expression levels of miR-155-5p in WT tissues and cells were both significantly down-regulated. Importantly, miR-155-5p was found to be involved in the malignant behavior of WT cells. MTT assay and flow cytometry assay demonstrated that miR-155-5p significantly inhibited the proliferation, caused stagnation of cells in G0/G1 phase, and promoted cell apoptosis. CREB1 was verified as a functional target gene of miR-155-5p, which was negatively regulated by miR-155-5p. Rescue experiments indicated that restoring the expression of CREB1 could interfere with the effects of miR-155-5p on WT cells. CONCLUSIONS: MiR-155-5p could regulate the proliferation, cell cycle and apoptosis of WT cells. These effects were achieved by regulating the expression of CREB1. Furthermore, our study might provide a new theoretical basis for the basic research of WT.

[Effects of electrospun collagen nanofibrous matrix on the biological behavior of human dental pulp cells].

OBJECTIVE: To compare cell adhesion, proliferation and odontoblastic differentiation of human dental pulp cells (hDPCs) on electrospun collagen nanofibrous matrix (Col_NFM) with that on collagen flat film (Col-FF), to investigate the biological effect of collagen nanofibrous matrix on hDPCs. METHODS: The surface morphology of the two different collagen scaffold was analyzed by scanning electron microscopy (SEM), and the contact angle and the swelling ratio were also measured. Then hDPCs were implanted on the two different collagen scaffolds, the cell morphology was observed using SEM and laser scanning microscope (LSM), and cell proliferation was evaluated by the CCK-8 assay. After hDPCs cultured on the two different collagen scaffold with odontoblastic medium for 14 days, the expression of odontoblastic differentiation related genes was detected by real-time PCR, and alizarin red staining was used to test the formation of mineralized nodules. RESULTS: From the SEM figures, the fibers' diameter of Col_NFM was (884±159) nm, and there were abundant three dimensional connected pore structures between the fibers of Col_NFM, while the surface of Col_FF was completely flat without pore structure. The contact angle at 0 s of Col_NFM was 85.03°±4.45°, and that of Col_FF was 98.98°±5.81°. The swelling ratio of Col_NFM was approximately 3 folds compared with dry weight sample, while that of Col_FF was just 1 fold. Thus Col_NFM indicated better hydrophilicity and swelling property. SEM and LSM showed that hDPCs on Col_NFM presented an irregular and highly branched phenotype, and could penetrate into the nanofibrous scaffold. In contrast, the cells were spread only on the surface of Col_FF with a spindle-shaped morphology. CCK-8 assays showed that hDPCs on Col_NFM showed higher proliferation rate than on Col_FF. After hDPCs were cultured on the two different collagen scaffolds with odontoblastic medium for 14 days, more expressions of odontoblastic differentiation related genes, such as dentin sialophosphoprotein (DSPP) and dentin matrix proten-1 (DMP1) were determined in Col_NFM group (P<0.05), and more mineralization depositions were also observed in Col_NFM group according to the results of alizarin red staining. CONCLUSION: Col_NFM with nanoscale microstructure achieves better hydrophilic and swelling properties than Col_FF, and hDPCs cultured with Col_NFM present higher activity on cell adhesion, proliferation and odontoblastic differentiation.

[Analysis on influencing factors of deaths from severe heat stroke in Shanghai, 2013-2017].

Objective: To explore the related factors of death from severe heat stroke in Shanghai from 2013 to 2017. Methods: The data of 1 152 patients with severe heat stroke who were divided into survival (n=1 037) and death (n=115) groups including gender, age and heat stroke type (heat cramp, heat exhaustion, heat apoplexy and the mixed type) were collected from meteorological bureau and case report system for high temperature heat stroke in Shanghai from 2013 to 2017. Meanwhile, the meteorological data of the onset date of severe heat stroke cases were collected, including maximum temperature, minimum temperature, daily temperature, relative humidity, air pressure, precipitation and wind speed. The differences of individual and meteorological factors between the two groups were compared, and multivariate logistic regression model was used to analyze the related factors of death from severe heat stroke. Results: Among 1 152 cases, the mean±SD of age was (56.29±18.95) years old, 843 (73.18%)were male, 962 (83.51%) were in the heat wave period; 322 cases (27.95%) were heat cramp, 170 cases (14.76%) were heat exhaustion, 533 cases (46.27%) were heat apoplexy and 114 cases (9.90%) were the mixed type. Daily average temperature ((32.81±1.99) ℃), daily maximum and minimum temperatures ((38.20±2.24) ℃ and (29.22±1.94) ℃) in survival group were lower than those in death group (all P values<0.001), which were (33.76±1.17) ℃, (39.19±1.31) ℃ and (29.72±1.66) ℃. Daily average relative humidity ((60.36±9.75)%) and daily minimum relative humidity ((41.26±9.71)%) in survival group were higher than those in death group(allP values <0.05), which were (54.59±6.89)% and (35.60±7.24)%. The results of logistic regression analysis suggested that compared with the cases with daily average humidity <60% and a mixed type heat stroke, the death OR (95%CI) values of cases with daily average humidity >60%, heat cramp, heat exhaustion and heat apoplexy were 0.31 (0.18,0.54), 0.13 (0.05,0.34), 0.68 (0.58,2.30) and 0.87 (0.48,1.58). Conclusion: The temperature, relative humidity and the type of heat stroke were the main related factors affecting the prognosis of severe heat stroke.

[A multicenter study of reference intervals for 15 laboratory parameters in Chinese children].

Objective: To establish comprehensive laboratory reference intervals for Chinese children. Methods: This was a cross-sectional multicenter study. From June 2013 to December 2014, eligible healthy children aged from 6-month to 17-year were enrolled from 20 medical centers with informed consent. They were assessed by physical examination, questionnaire survey and abdominal ultrasound for eligibility. Fasting blood samples were collected and delivered to central laboratory. Measurements of 15 clinical laboratory parameters were performed, including estradiol (E2), testosterone(T), luteinizing hormone(LH), follicle-stimulating hormone(FSH), alanine transaminase(ALT), serum creatinine(Scr), cystatin C, immunoglobulin A(IgA), immunoglobulin G(IgG), immunoglobulin M(IgM), complement (C3, C4), alkaline phosphatase(ALP), uric acid(UA) and creatine kinase(CK). Reference intervals were established according to central 95% confidence intervals for reference population, stratified by age and sex. Results: In total, 2 259 children were enrolled. Finally, 1 648 children were eligible for this study, including 830 boys and 818 girls, at a mean age of 7.4 years. Age- and sex- specific reference intervals have been established for the parameters. Reference intervals of sex hormones increased gradually with age. Concentrations of ALT, cystatin C, ALP and CK were higher in children under 2 years old. Serum levels of sex hormones, creatinine, immunoglobin, CK, ALP and urea increased rapidly in adolescence, with significant sex difference. In addition, reference intervals were variable depending on assay methods. Concentrations of ALT detected by reagents with pyridoxal 5'-phosphate(PLP) were higher than those detected by reagents without PLP. Compared with enzymatic method, Jaffe assay always got higher results of serum creatinine, especially in children younger than 9 years old. Conclusion: This study established age- and sex- specific reference intervals, for 15 clinical laboratory parameters based on defined healthy children.

[Study on the current situation of China's First List of Rare Diseases based on 15 million hospitalizations].

Objective: To investigate and analyze the distribution of 121 diseases of China's First List of Rare Diseases based on hospitalized patients of tertiary hospitals and to explore the current situation of rare diseases in China. Methods: Based on previous data of study from Beijing Society of Rare Diseases, a comparison between China's First List of Rare Diseases and the survey list from the pre-study was performed. Descriptive analysis was carried out on the current situation of rare diseases on hospitalizations in 96 tertiary hospitals from year of 2014 to 2015. Results: Nineteen out of 121 diseases on China's First List of Rare Diseases were not included in the rare diseases survey list of Beijing Society of Rare Diseases. The total number of other 102 rare disease cases was 54 468, accounting for 0.35% of the inpatients during the same period. The top ten most and least cases with rare disease were demonstrated in this study. The number of the top ten most cases was 37 977, accounting for 0.25% of the inpatients during the same period. The number of the top ten least cases was 24, accounting for 0.000 16% of the inpatients during the same period. The top most five types of rare diseases counted on the provinces and municipalities were Beijing, Hunan, Shanghai, Shandong and Guangdong. The top five most cases of rare diseases counted on the provinces and municipalities were Beijing, Shanghai, Guangdong, Shandong and Hubei. The age distribution showed that the cases with rare diseases aged 25-64 years accounted for 45.8%, and the cases in children aged 0-14 accounted for 28.6%. The top ten readmission rate ranged from 28.42% to 64.88%. Conclusions: This study preliminarily investigates the number, type, province and municipality distribution, age distribution, and readmission rate of 121 rare diseases from China's First List of Rare Diseases in the hospitalized patients of tertiary hospitals, which provides important data for registration study, medical and drug policy making and other relevant work on rare diseases in China in the future.

[Acute effect of ambient air pollution on small airway lung functions among school children in Shanghai].

Objective: To study the acute effects of compound ambient air pollution on small airway lung functions among school children in Shanghai. Method: A longitudinal survey on lung functions was conducted among 233 school-children from three schools (A, B and C, located in innerring, mid-ring and outer-ring areas). Lung function test was performed once a week for 3 times respectively, among children in school A and B in Dec. 2013 and in school C in Dec. 2014. The fourth lung function test was tested in Jun. 2014 and May 2015 in the respective schools. Results: from the lung function would include items as: forced mid-expiratory flow at 25% of forced vital capacity (MEF(25%)), mid-expiratory flow at 50% of forced vital capacity (MEF(50%)), mid-expiratory flow at 75% of forced vital capacity (MEF(75%)) and mid-expiratory flow between 25% and 75% of the forced vital capacity (FEF(25%-75%)). Data regarding the daily air quality real-time of PM(2.5), PM(10), SO(2) and NO(2) in Dec. 2013, Dec. 2014, Jun. 2014 and May. 2015 from the three environmental monitoring spots and meteorological data from the Shanghai Meteorological Service system which were physically close to the three schools, were collected simultaneously. Linear mixed effect model was used to examine the levels of correlation between lung function indicators and ambient air pollutants. Results When confounding factors on meteorology and individuals were controlled, the lag effects and accumulated lag effects were found to have existed between the internal quarter rang (IQR) concentration of PM(2.5) and PM(10) in lag2 day and lag02 days, IQR concentration of SO(2) in lag02 day and IQR concentration of NO(2) lag0 day, when small airway lung functions like MEF(25%), MEF(50%), MEF(75%) and FEF(25%-75%)(P<0.05) were inspected. Results from the two air pollutants model analysis showed that SO(2) and NO(2) presenting interactive effects with PM(2.5), PM(10) and lag effects more significant than the individual SO(2) and NO(2), respectively (P<0.05). Conclusion: Contents on the ambient air pollutants as PM(2.5), PM(10), SO(2) and NO(2) were negatively associated with the lung functions in the small airways of children, in Shanghai.

Meta-analysis of TAFI polymorphisms and risk of cardiovascular and cerebrovascular diseases.

Cardiovascular and cerebrovascular diseases (CCVDs) are common and have high rates of morbidity, mortality, and recurrence. Thrombin-activatable fibrinolysis inhibitor (TAFI) is also known as carboxypeptidase B2 and is encoded by the CPB2 gene; CPB2 polymorphisms have been explored in a variety of studies, but their correlation to the risk of CCVDs remains ambiguous. We examined the hypothesized associations between CPB2 mutations and CCVDs in a general population. We searched, Embase, the Cumulative Index to Nursing and Allied Health Literature, the Science Citation Index, and several Chinese databases without applying any language restrictions. Nine case-control studies were analyzed in the current meta-analysis, and odds ratios (ORs) with their 95% confidence intervals were calculated. The pooled ORs indicated that the CPB2 rs3742264 G>A polymorphism was associated with an increased risk of CCVDs in the allele model (all P values < 0.05). A similar result for the CPB2 rs1926447 C>T polymorphism and CCVDs risk was detected in the allele model (P < 0.05). Ethnicity subgroup analysis implied that the rs3742264 G>A polymorphism was more likely to lead to the development of cerebrovascular disease in Asians (all P values < 0.05), whereas rs1926447 C>T was associated with cardiovascular disease among Africans (all P values < 0.05). These data suggest that the polymorphisms investigated, especially rs3742264 G>A and rs1926447 C>T, have a modest effect on susceptibility to CCVDs.

Role of IL-10 polymorphisms in susceptibility to hepatitis B virus-related hepatocellular carcinoma.

We conducted a case-control study to investigate the role of three common single nucleotide polymorphisms of IL-10 (-592G/A, -819T/C, and -1082A/C) in the development of hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC). The study included 173 HBV-related HCC patients and 182 healthy controls. A polymerase chain reaction-restriction fragment length polymorphism assay was applied to assess the sequence variants of interest. Compared with control subjects, HCC patients were more likely to be older (t = 1.94, P = 0.03), have a family history of cancer (chi square = 17.86, P < 0.001), and exhibit higher alanine transaminase (t = 13.32, P < 0.001) and aspartate transaminase (t = 12.63, P < 0.001) levels. Using unconditional logistic regression analyses, we found that the GG genotype of -592G/A was associated with increased risk of HCC [odds ratio (OR) = 2.20, 95% confidence interval (CI) = 1.12-4.38], compared to the AA genotype. Under a dominant model, the AG+GG genotype correlated with HBV-related HCC susceptibility compared to the AA genotype, with an OR (95%CI) of 1.56 (1.02-2.48). Moreover, a recessive model showed the GG genotype to be associated with elevated risk of HCC compared to the AA+AG genotype (OR = 1.85, 95%CI = 1.01-3.47). However, no significant association between the -819T/C and -1082A/C variants and development of HBV-related HCC was observed under codominant, dominant, and recessive models. We conclude that the IL-10 -592G/A polymorphism does play a role in susceptibility to HBV-related HCC under codominant, dominant, and recessive models.

Alterations in microRNA expression profile in rabies virus-infected mouse neurons.

Rabies virus (RABV) is known to cause a fatal infection in many mammalian species, yet its pathogenesis remains poorly understood. This study was performed to analyze the microRNA (miRNA) expression profiles in RABV-infected primary neurons of mice. A total of 53 miRNAs were found to be differentially expressed in RABV-infected samples compared with mock samples in a time-dependent manner. Among them, the expression of ten miRNAs was validated by real-time RT-PCR. Potential target genes of differentially expressed miRNAs were predicted by TargetScan. Further bioinformatics analysis indicated that these predicted targets were overrepresented in neuronal function-related Gene Ontology (GO) terms and biological pathways. The results of this study suggest that RABV may cause neuronal dysfunction by regulating cellular miRNA expression.

Photo-induced processes in collagen-hypericin system revealed by fluorescence spectroscopy and multiphoton microscopy.

Collagen is the main structural protein and the key determinant of mechanical and functional properties of tissues and organs. Proper balance between synthesis and degradation of collagen molecules is critical for maintaining normal physiological functions. In addition, collagen influences tumor development and drug delivery, which makes it a potential cancer therapy target. Using second harmonic generation, two-photon excited fluorescence microscopy, and spectrofluorimetry, we show that the natural pigment hypericin induces photosensitized destruction of collagen-based tissues. We demonstrate that hypericin-mediated processes in collagen fibers are irreversible and may be used for the treatment of cancer and collagen-related disorders.

Pharmacokinetic studies of bergapten in dog plasma by using a LC-MS/MS method studies.

A sensitive LC-MS/MS method was developed for the determination of bergapten in dog plasma. The chromatographic separation was carried out on a Hypersil ODS column with a mobile phase consisting of methanol-water. The plasma sample was precipitated with methanol and prepare for injecting onto the LC-MS/MS system. The detection was performed on a triple quadrupole tandem mass spectrometer by MRM via electro spray ionization source. The standard curve for bergapten was linear over the concentration range of 0.5-500 ng/mL with a lower limit of quantification of 0.5 ng/mL. The inter-day and intra-day precision (R.S.D.%) for bergapten varied between 3.4 and 11.5. The corresponding inter-day and intra-day accuracy (Bias%) ranged between -3.8 and 6.9. For the pharmacokinetic analysis of serum, the mean (SD) values obtained for the bergapten were as follows: Cmax, 228.5 (14.3) ng/ml; Tmax, 4.2 (0.4) h; t1/2, 6.9 (2.3) h; AUC0-t h, 2507.2 (168.5) ng · h/mL and AUC0-∞, 3 219.2 (211.4) ng · h/mL, respectively.

A simple and sensitive HPLC-UV method for the determination of swainsonine in rat plasma and its application in a pharmacokinetic study.

A simple, rapid, selective and sensitive HPLC-UV method was developed and validated for the determination of swainsonine (SWSN) in rat plasma. The analyte was extracted from rat plasma with methanol as the extraction solvent. The LC separation was performed on a Diamonsil® C18 (250×4.6 mm, 5 µm) analytical column with a mobile phase consisting of acetonitrile-potassium dihydrogen phosphate (25 mmol/l, pH=7.5) at a flow rate of 1.0 ml/min. There was a good linearity over the range of 10-500 ng/ml (r=0.9995) with a weighted (1/C2) least square method. The lower limit of quantification was proved to be 10 ng/ml. The accuracy was within 4.8% in terms of relative error and the intra- and inter-day precisions were less than 9.0% in terms of relative standard deviation. After validation, the method was successfully applied to characterize the pharmacokinetics of SWSN in rats.

Tumor-targeted gene therapy using Adv-AFP-HRPC/IAA prodrug system suppresses growth of hepatoma xenografted in mice.

Clinical efficacy of current therapies for hepatocellular carcinoma (HCC) treatment is limited. Indole-3-acetic acid (IAA) is non-toxic for mammalian cells. Oxidative decarboxylation of IAA by horseradish peroxidase (HRP) leads to toxic effects of IAA. The purpose of this study was to investigate the effects of a novel gene-targeted enzyme prodrug therapy with IAA on hepatoma growth in vitro and in vivo mouse hepatoma models. We generated a plasmid using adenovirus to express HRP isoenzyme C (HRPC) with the HCC marker, alpha-fetoprotein (AFP), as the promoter (pAdv-AFP-HRPC). Hepatocellular cells were infected with pAdv-AFP-HRPC and treated with IAA. Cell death was detected using MTT assay. Hepatoma xenografts were developed in mice by injection of mouse hepatoma cells. The size and weight of tumors and organs were evaluated. Cell death in tumors was assessed using hematoxylin and eosin-stained tissue sections. HRPC expression in tissues was detected using Reverse Transcriptase-Polymerase Chain Reaction. IAA stimulated death of hepatocellular cells infected with pAdv-AFP-HRPC, in a dose- and time-dependent manner, but not in control cells. Growth of hepatoma xenografts, including the size and weight, was inhibited in mice treated with pAdv-AFP-HRPC and IAA, compared with that in control group. pAdv-AFP-HRPC/IAA treatment induced cell death in hepatoma xenografts in mice. HRPC gene expressed only in hepatoma, but not in other normal organs of mice. pAdv-AFP-HRPC/IAA treatment did not cause any side effects on normal organs. These findings suggest that pAdv-AFP-HRPC/IAA enzyme/prodrug system may serve as a strategy for HCC therapy.

Apical membrane rupture and backward bile flooding in acetaminophen-induced hepatocyte necrosis.

Morphological changes of hepatocyte death have so far only been described on cells in culture or in tissue sections. Using a high-resolution and high-magnification multiphoton microscopic system, we recorded in living mice serial changes of acetaminophen (APAP)-induced hepatocyte necrosis in relevance to metabolism of a fluorogenic bile solute. Initial changes of hepatocyte injury included basal membrane disruption and loss of mitochondrial membrane potential. An overwhelming event of rupture at adjacent apical membrane resulting in flooding of bile into these hepatocytes might ensue. Belbs formed on basal membrane and then dislodged into the sinusoid circulation. Transmission electron microscopy disclosed a necrotic hepatocyte depicting well the changes after apical membrane rupture and bile flooding. Administration of the antidote N-acetylcysteine dramatically reduced the occurrence of apical membrane rupture. The present results demonstrated a hidden but critical step of apical membrane rupture leading to irreversible APAP-induced hepatocyte injury.

Differential expression profiling of ovarian genes in prelaying and laying geese.

The identification of the differential expression of genes in the ovaries of egg-laying and prelaying Zi geese is required to improve the laying performance of the geese. In the present study, suppression subtractive hybridization and reverse dot-blot were employed to identify such genes, using the ovary as a model. Furthermore, expression profiling of estrogen receptor 1, estrogen receptor 2, follicle stimulating hormone receptor, prolactin receptor, ferritin H chain, and ovary differentially expressed unknown gene 08 in ovaries from geese was performed by quantitative real-time PCR. Total RNA from the ovaries of laying and prelaying Zi geese was pooled and the mRNA was isolated. The cDNA that was reverse-transcribed from the ovarian mRNA of the prelaying geese was subtracted from the cDNA isolated from the laying geese. Four hundred sixty-five clones containing putative differentially expressed gene fragments were further identified by reverse dot-blot. Ninety-seven clones were subjected to sequencing and further analysis. Sequence analysis showed that the expression of 18 known (including a mitochondrial gene) and 8 unknown gene fragments was higher in the ovaries of laying geese compared with prelaying geese. Seventeen of the known genes encode proteins that belong to groups involved with binding, catalytic activity, enzyme regulatory activity, signal transducer activity, structural molecule, and transporter activity. The results of the quantitative real-time PCR showed that the expression of estrogen receptor 1, estrogen receptor 2, follicle stimulating hormone receptor, prolactin receptor, ferritin H chain, and ovary differentially expressed unknown gene 08 was higher in the ovaries of the laying geese than in those of the prelaying geese (P<0.05). These differentially expressed genes may be relevant to the progression of prelaying geese to the egg-laying stage. Further study is required to elucidate the molecular mechanism that controls egg-laying in geese, to improve the productivity of laying geese.