Cytomegalovirus Latency Exacerbated Small-for-size Liver Graft Injury Through Activation of CCL19/CCR7 in Hepatic Stellate Cells.

BACKGROUND: The interplay between cytomegalovirus (CMV) latency and graft malfunction after living donor liver transplantation remains poorly defined because of the complexity of clinical confounding factors. Here, we aimed to investigate the effects of CMV latency on small-for-size graft injury and to get further insight into the pathogenic role of hepatic stellate cells (HSCs) in this process. METHODS: Rat orthotopic liver transplantation with small-for-size grafts was performed in a CMV latent model developed in immunocompetent Sprague Dawley rats using Priscott strain. Posttransplant graft injury including hepatocyte damage, stellate cell activation, and fibrogenesis was evaluated. Differential gene expression of HSCs in response to CMV latency was screened by cDNA microarray. Clinical validation was further conducted in human biopsies. RESULTS: CMV latency aggravated hepatocyte apoptosis/necrosis in the early phase and enhanced HSC expansion and graft fibrosis during the middle-late phase in small-for-size liver grafts of the rat model. cDNA microarray mining revealed CCL19/CCR7 as one of the most noteworthy pathways bridging HSC activation and liver graft injury in the presence of CMV latency. Together with CCL19 upregulation, coherent overexpression of CCR7 in accumulated HSCs was confirmed in both rat and human CMV latent recipients. Moreover, addition of CCL19 in vitro promoted HSC migration by increasing the level of matrix metalloproteinase-2. CONCLUSIONS: Our data demonstrated that CMV latency aggravated early/late phase liver graft damage and fibrogenesis via CCL19/CCR7/HSCs axis. Blockade of CMV latency-related stellate cell activation may shed light on the strategy of graft protection clinically.

Plasma miRNA-122-5p and miRNA-151a-3p identified as potential biomarkers for liver injury among CHB patients with PNALT.

BACKGROUND: Plasma microRNA (miRNA) levels may be altered during pathological processes; therefore, they may potentially serve as biomarkers for the diagnosis and prognosis of human diseases. This study aimed to explore whether plasma miRNAs may serve as new biomarkers for liver injury among chronic hepatitis B (CHB) patients with normal or nearly normal alanine aminotransferase (ALT) levels. METHODS: Plasma miRNAs from each of three independent groups (no prominent liver injury and persistently normal ALT levels, NPNALT; significant liver injury with persistently normal ALT levels, SPNALT; and healthy) were profiled by miRNA microarray analysis. Differentially expressed miRNAs were then validated by a quantitative reverse transcription polymerase chain reaction (qRT-PCR) assay. The area under the receiver operating characteristic (AUC) curve was used to analyze the candidate miRNAs validated by qRT-PCR for diagnostic accuracy. RESULTS: Twenty differentially expressed miRNAs were identified by microarray analysis. Seven miRNAs with elevated serum levels were validated by qRT-PCR analysis, and four of them were significantly different between the SPNALT and NPNALT groups. The AUCs of hsa-miR-122-5p and hsa-miR-151-3p were 0.877 (cutoff value = 13.38; 95% CI 0.792-0.963; sensitivity = 83.3%, specificity = 80%) and 0.882 (cutoff value = 7.4; 95% CI 0.797-0.966; sensitivity = 83.3%, specificity = 73.3%), respectively, indicating early liver injury. However, there was no significant correlation of miRNAs with either necroinflammation or fibrosis. CONCLUSION: Serum hsa-miR-122-5p and hsa-miR-151-3p may function as new biomarkers for liver injury in SPNALT patients. With these two biomarkers, we may be able to identify a subset of patients who are experiencing liver injury but have normal ALT levels for further evaluation with a biopsy procedure.

Non-ALT biomarkers for markedly abnormal liver histology among Chinese persistently normal alanine aminotransferase-chronic hepatitis B patients.

AIM: To determine incidence and clinical biomarkers of marked necroinflammation and fibrosis characteristics among chronic hepatitis B (CHB) patients with persistently normal alanine aminotransferase (PNALT). METHODS: Liver biopsy was performed on 115 CHB patients with PNALT. Necroinflammation and fibrosis were graded by the Knodell histologic activity index and the Ishak fibrosis score, respectively. Correlations between the available clinical parameters and necroinflammation and fibrosis were analysed. RESULTS: Marked necroinflammation (Knodell activity index ≥ 7) and fibrosis (Ishak fibrosis score ≥ 3) were found in 36.5% and 15.5% of CHB patients with PNALT, respectively. Following a univariate logistic regression analysis, multiple logistic regression analysis indicated that aspartate transaminase (AST) (AUROC = 0.852, cut-off value = 22.5 U/L) serves as an independent predictor of notable liver inflammation, while platelet (PLT) count (AUROC = 0.905, cut-off value = 171.5 ×109/mL) and gamma-glutamyl transpeptidase (GGT) (AUROC = 0.909, cut-off value = 21.5 U/L) level serve as independent predictors of notable liver fibrosis. CONCLUSION: A considerable proportion of marked histological abnormalities existed in our cohort, who will benefit from optimal therapeutic strategies administered according to predictive indication by AST, PLT and GGT levels.

Hyaluronic acid reagent functional chitosan-PEI conjugate with AQP2-siRNA suppressed endometriotic lesion formation.

To identify a new drug candidate for treating endometriosis which has fewer side effects, a new polymeric nanoparticle gene delivery system consisting of polyethylenimine-grafted chitosan oligosaccharide (CSO-PEI) with hyaluronic acid (HA) and small interfering RNA (siRNA) was designed. There was no obvious difference in sizes observed between (CSO-PEI/siRNA)HA and CSO-PEI/siRNA, but the fluorescence accumulation in the endometriotic lesion was more significant for (CSO-PEI/siRNA)HA compared with CSO-PEI/siRNA due to the specific binding of HA to CD44. In addition, the (CSO-PEI/siRNA)HA nanoparticle gene therapy significantly decreased the endometriotic lesion sizes with atrophy and degeneration of the ectopic endometrium. The epithelial cells of ectopic endometrium from rat models of endometriosis showed a significantly lower CD44 expression than control after treatment with (CSO-PEI/siRNA)HA. Furthermore, observation under an electron microscope showed no obvious toxic effect on the reproductive organs. Therefore, (CSO-PEI/siRNA)HA gene delivery system can be used as an effective method for the treatment of endometriosis.

[Expression of hTERT and Mad1 in lung cancer in Gejiu and Xuanwei of Yunnan Province].

OBJECTIVE: To study the expression of hTERT mRNA and Mad1 protein in lung cancer of Gejiu and Xuanwei and normal lung tissue and to investigate their correlations with lung cancer. METHODS: Mad1 protein was detected by immunohistochemistry S-P method, and hTERT message RNA (mRNA) was detected by in situ hybridization (ISH) in 40 specimens of lung cancer of Gejiu Tin miners and 20 specimens of lung cancer of Xuanwei peasants and 20 specimens of normal lung tissue. The positive signals were quantitatively analyzed by HPIAS-100. RESULTS: The positive unit (PU) of Mad1 protein was 16.77 +/- 6.01 in Gejiu Tin Miners lung cancer group, and 19.36 +/- 4.54 in Xuanwei peasant lung cancer group, compared with the normal lung tissue (46.05 +/- 7.26). The difference was highly significant (P < 0.01); The PU of hTERT mRNA was 72.10 +/- 13.07 in Gejiu Tin miners lung cancer group, and 74.20 +/- 15.17 in Xuanwei peasant lung cancer group, which was higher than that in normal tissue group (10.70 +/- 2.21). The difference was significant (P < 0.01). The expression of Mad1 protein was negatively correlated with the expression hTERT mRNA (P < 0.05, r = 0.9881, r = -0.999). CONCLUSION: Reduced hTERT mRNA expression may play an important role in the occurrence of lung cancer. The expression of hTERT mRNA and deletion of Mad1 protein are closely related to pathogenesis of lung cancer.

[Loss of exons of FHIT gene and FHIT protein expression in Yunnan tin miners with lung cancer].

BACKGROUND & OBJECTIVE: Yunnan tin miners have an extremely high incidence of lung cancer. Abnormality of the fragile histidine triad (FHIT) gene has been proved to closely relate to lung cancer development. This study was to explore the loss of exons 3, 4, 5 and 8 of FHIT gene and FHIT protein expression in Yunnan Tin miners with lung cancer. METHODS: The exons 3, 4, 5, and 8 of FHIT gene in lung cancer cell line YTMCC of a Yunnan Tin miner, and 30 specimens of lung cancer from Yunnan Tin miners in Gejiu district and 22 specimens of lung cancer from non-miners in other regions were detected by polymerase chain reaction (PCR). The expression of FHIT protein in 90 specimens of human lung cancer and 43 specimens of lung cancer developed by the intratracheal instillation of Yunnan Tin mineral dust in F344 rats was detected by immunohistochemistry. RESULTS: Losses of exon 3 and exon 8 were detected in YTMCC cells. Lung cancer samples from Yunnan Tin miners and non-miners exhibited heterozygous loss of FHIT gene among exons 3, 4, 5 and 8. The percentages of FHIT gene deletion and loss of FHIT protein expression in Yunnan Tin miners with lung cancer were 68.2% and 70.6%, respectively. The percentages of FHIT gene heterozygous loss was significantly higher in lung cancer tissue than in normal lung tissue (P < 0.01). Loss of FHIT protein expression was detected in the early stage of F344 rat lung canceration after treatment of Yunnan tin Mineral dust:the percentage was 100% in 8 specimens of squamous dysplasia, carcinoma in situ, and early stage of squamous cell carcinoma. CONCLUSIONS: Deletion and abnormal expression of FHIT gene are common in Yunnan tin miners and non-miners with lung cancer. The high rate of loss of FHIT expression in precancerous lesions and lung cancer at early stage indicates that FHIT could be an early screening target of lung cancer.