FXR activation remodels hepatic and intestinal transcriptional landscapes in metabolic dysfunction-associated steatohepatitis.

The escalating obesity epidemic and aging population have propelled metabolic dysfunction-associated steatohepatitis (MASH) to the forefront of public health concerns. The activation of FXR shows promise to combat MASH and its detrimental consequences. However, the specific alterations within the MASH-related transcriptional network remain elusive, hindering the development of more precise and effective therapeutic strategies. Through a comprehensive analysis of liver RNA-seq data from human and mouse MASH samples, we identified central perturbations within the MASH-associated transcriptional network, including disrupted cellular metabolism and mitochondrial function, decreased tissue repair capability, and increased inflammation and fibrosis. By employing integrated transcriptome profiling of diverse FXR agonists-treated mice, FXR liver-specific knockout mice, and open-source human datasets, we determined that hepatic FXR activation effectively ameliorated MASH by reversing the dysregulated metabolic and inflammatory networks implicated in MASH pathogenesis. This mitigation encompassed resolving fibrosis and reducing immune infiltration. By understanding the core regulatory network of FXR, which is directly correlated with disease severity and treatment response, we identified approximately one-third of the patients who could potentially benefit from FXR agonist therapy. A similar analysis involving intestinal RNA-seq data from FXR agonists-treated mice and FXR intestine-specific knockout mice revealed that intestinal FXR activation attenuates intestinal inflammation, and has promise in attenuating hepatic inflammation and fibrosis. Collectively, our study uncovers the intricate pathophysiological features of MASH at a transcriptional level and highlights the complex interplay between FXR activation and both MASH progression and regression. These findings contribute to precise drug development, utilization, and efficacy evaluation, ultimately aiming to improve patient outcomes.

Bacterial Microbiome Differences between the Roots of Diseased and Healthy Chinese Hickory (Carya cathayensis) Trees.

Carya cathayensis is an important economic nut tree that is endemic to eastern China. As such, outbreaks of root rot disease in C. cathayensis result in reduced yields and serious economic losses. Moreover, while soil bacterial communities play a crucial role in plant health and are associated with plant disease outbreaks, their diversity and composition in C. cathayensis are not clearly understood. In this study, Proteobacteria, Acidobacteria, and Actinobacteria were found to be the most dominant bacterial communities (accounting for approximately 80.32% of the total) in the root tissue, rhizosphere soil, and bulk soil of healthy C. cathayensis specimens. Further analysis revealed the abundance of genera belonging to Proteobacteria, namely, Acidibacter, Bradyrhizobium, Paraburkholderia, Sphaerotilus, and Steroidobacter, was higher in the root tissues of healthy C. cathayensis specimens than in those of diseased and dead trees. In addition, the abundance of four genera belonging to Actinobacteria, namely, Actinoallomurus, Actinomadura, Actinocrinis, and Gaiella, was significantly higher in the root tissues of healthy C. cathayensis specimens than in those of diseased and dead trees. Altogether, these results suggest that disruption in the balance of these bacterial communities may be associated with the development of root rot in C. cathayensis, and further, our study provides theoretical guidance for the isolation and control of pathogens and diseases related to this important tree species.

Exogenous citrate restores the leaf metabolic profiles of navel orange plants under boron deficiency.

Boron (B) is an essential micronutrient for higher plants, and its deficiency causes a change in the citrate concentration in leaves of young navel orange plants. Although citrate has been implicated in the regulation of gene expression for many transcripts, it is unclear whether citrate can affect metabolic profiling under B deficiency and if so, how many metabolites are affected. In this study, GC-TOF-MS-based untargeted metabolite profiling was used to identify the physiological effects of exogenous citrate on recovery of metabolites in B-deficient orange plants. There were 31 increased and 24 decreased metabolites in the boron-deficient (BD) group leaves relative to those of the boron-adequate (BA) group. Boron deficiency-induced changes in many metabolites were restored to the level of BA (control) group leaves or showed a recovery tendency at 1 week after citrate supply (foliar application of citrate, BDFC), including 11 organic acids, 9 sugars and polyols, 10 amino acids, and 4 other compounds. To compare with the metabolic recovery effects of exogenous citrate on B deficiency, exogenous application of B (borate) was also performed under same conditions (BDFB), and similar effects on the regulation of metabolic homeostasis under B deficiency were observed. Both the results of principal component analysis (PCA) and hierarchical cluster analysis (HCA) showed that BA, BDFC, and BDFB were relatively similar and clustered close to each other. There are different responsive and regulatory mechanisms to the additions of exogenous citrate in navel orange leaves under B adequate and deficient conditions. Based on these results, we suggest that citrate is an important component of the B deficiency stress response, and exogenous application of citrate generally restores the leaf metabolic profiles of navel orange plants under boron deficiency, which might play a positive role in this stress tolerance.

Genome-Wide Identification of Long Non-coding RNA in Trifoliate Orange (Poncirus trifoliata (L.) Raf) Leaves in Response to Boron Deficiency.

Long non-coding RNAs (lncRNAs) play important roles in plant growth and stress responses. As a dominant abiotic stress factor in soil, boron (B) deficiency stress has impacted the growth and development of citrus in the red soil region of southern China. In the present work, we performed a genome-wide identification and characterization of lncRNAs in response to B deficiency stress in the leaves of trifoliate orange (Poncirus trifoliata), an important rootstock of citrus. A total of 2101 unique lncRNAs and 24,534 mRNAs were predicted. Quantitative real-time polymerase chain reaction (qRT-PCR) experiments were performed for a total of 16 random mRNAs and lncRNAs to validate their existence and expression patterns. Expression profiling of the leaves of trifoliate orange under B deficiency stress identified 729 up-regulated and 721 down-regulated lncRNAs, and 8419 up-regulated and 8395 down-regulated mRNAs. Further analysis showed that a total of 84 differentially expressed lncRNAs (DELs) were up-regulated and 31 were down-regulated, where the number of up-regulated DELs was 2.71-fold that of down-regulated. A similar trend was also observed in differentially expressed mRNAs (DEMs, 4.21-fold). Functional annotation of these DEMs was performed using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses, and the results demonstrated an enrichment of the categories of the biosynthesis of secondary metabolites (including phenylpropanoid biosynthesis/lignin biosynthesis), plant hormone signal transduction and the calcium signaling pathway. LncRNA target gene enrichment identified several target genes that were involved in plant hormones, and the expression of lncRNAs and their target genes was significantly influenced. Therefore, our results suggest that lncRNAs can regulate the metabolism and signal transduction of plant hormones, which play an important role in the responses of citrus plants to B deficiency stress. Co-expression network analysis indicated that 468 significantly differentially expressed genes may be potential targets of 90 lncRNAs, and a total of 838 matched lncRNA-mRNA pairs were identified. In summary, our data provides a rich resource of candidate lncRNAs and mRNAs, as well as their related pathways, thereby improving our understanding of the role of lncRNAs in response to B deficiency stress, and in symptom formation caused by B deficiency in the leaves of trifoliate orange.

Ligand-induced activation of ERK1/2 signaling by constitutively active Gs-coupled 5-HT receptors.

5-HT4R, 5-HT6R, and 5-HT7AR are three constitutively active Gs-coupled 5-HT receptors that have key roles in brain development, learning, memory, cognition, and other physiological processes in the central nervous system. In addition to Gs signaling cascade mediated by these three 5-HT receptors, the ERK1/2 signaling which is dependent on cyclic adenosine monophosphate (cAMP) production and protein kinase A (PKA) activation downstream of Gs signaling has also been widely studied. In this study, we investigated these two signaling pathways originating from the three Gs-coupled 5-HT receptors in AD293 cells. We found that the phosphorylation and activation of ERK1/2 are ligand-induced, in contrast to the constitutively active Gs signaling. This indicates that Gs signaling alone is not sufficient for ERK1/2 activation in these three 5-HT receptors. In addition to Gs, we found that ß-arrestin and Fyn are essential for the activation of ERK1/2. Together, these results put forth a novel mechanism for ERK1/2 activation involving the cooperative action of Gs, ß-arrestin, and Fyn.

Occurrence and distribution of phosphorus fractions in sediments of Liangzi Lake under typical hydrodynamic conditions.

Understanding the transformation and chronological accumulation of phosphorus (P) forms under typical hydrodynamic conditions of a lake is important for clarifying the process of lake evolution and eutrophication. The occurrence and distribution of sediment P fractions (total, TP; inorganic, IP; and organic, OP), phytate content, and phytase activity at different profile depths (0-8 m) and parent material ages (0.8-11 ka BP) were examined at different ecological locations (inlet, outlet, and center) of the freshwater Liangzi Lake in Hubei Province, China. Sediment P-forms at locations of different hydrodynamic conditions increased from the inlet to the outlet. IP constituted ∼40-71% of TP, whereas the OP content was generally lower in the sediment. The two forms of IP extracted by HCl and NaOH varied quantitatively with depth and location: HCl-P ≈ NaOH-P (above 0.8 m) or HCl-P > NaOH-P (below 0.8 m) at the inlet; HCl-P > NaOH-P (above 0.8 m) and HCl-P ≈ NaOH-P (below 0.8 m) at the outlet; and HCl-P < NaOH-P at the center of the lake. Compared with labile and moderately resistant OP, moderately labile OP exhibited substantial quantitative changes and occurred at high levels. The variation trend in the phytate content coincided with that of TP, whereas phytase activity varied inversely with location. Low levels of P forms occurred in the sediment below 4.5 m and before 8.6 ka BP, consistent with the oligotrophic period of the lake. During 2-4 ka BP, the P forms first increased rapidly and then stabilized thereafter. From that time period until modern times, TP and phytate increased, whereas IP and OP decreased significantly. The results indicate that the hydrodynamic conditions of the water bodies and the sediments of different ages strongly influenced the occurrence and distribution of sediment P forms, and the sediment TP and phytate contents would be candidate indices to reflect the P input and eutrophication history of freshwater lakes.

[Four alien medical mollusk found in Dapeng Peninsula, Shenzhen City].

OBJECTIVE: To investigate the alien snails in Dapeng Peninsula, Shenzhen City. METHODS: The survey on the snail diversity in Dapeng Peninsula was carried out from August 2012 to Jan 2013, and the species of the alien snails were identified according to the shell morphology. RESULTS: Four species of alien snails including Achatinafulica, Pomacea canaliculata, Physa acuta, and Biomphalaria straminea were found in Dapeng Peninsula, P. acuta was found in all of the collected sites, A. fulica and P. canaliculata were distributed in five regions except Yangmeikeng area, and B. straminea was just found in Dapeng Town. CONCLUSION: Four species of important alien snails invade widely in Dapeng Peninsula, Shenzhen City. As their potential risk to the disease transmission and agriculture production, the relative departments should strengthen the control and prevention.

Induced apoptosis of osteoblasts proliferating on polyhydroxyalkanoates.

The mechanism study on behaviors of cells influenced by biomaterial surface properties can provide profound guidances for functional tissue engineering scaffolds design. In this study, regulation of integrin-mediated cell-substrate interactions using rat osteoblasts incubated on PHA films was investigated. Compared with tissue culture plate (TCP), poly-3-hydroxybutyrate (PHB), copolymer of 3-hydroxybutyrate and 3-hydroxyvalerate (PHBV) and copolymer of 3-hydroxybutyrate and 3-hydroxyhexanoate (PHBHHx), osteoblasts inoculated on a terpolymer of 3-hydroxybutyrate, 3-hydroxyvalerate and 3-hydroxyhexanoate (PHBVHHx) were found to have higher apoptosis rates. Several integrin subunits in osteoblasts grown on PHBVHHx showed altered expressions. Simultaneously, extracellular matrics (ECM) were also remodeled on the material surface. Osteoblasts showed a higher expression of integrin subunit ß3 and αv on PHBVHHx films compared with that on TCP. On the other hand, less vitronectin, osteopontin and fibronectin, the main ligands for integrin ß3 were expressed and deposited in ECM. The unligated integrin ß3 could recruit caspase-8 to the membrane and activate its downstream signaling which was proven by the caspase-8 activation assay. It was therefore concluded that the induced apoptosis of osteoblasts on PHBVHHx was regulated by recruitment of caspase-8 to the unligated integrin ß3.

An assessment of the risks of carcinogenicity associated with polyhydroxyalkanoates through an analysis of DNA aneuploid and telomerase activity.

Polyhydroxyalkanoates (PHA) are aliphatic polyesters synthesized by many bacteria. Because of their flexible mechanical strengths, superior elastic property, biodegradability and biocompatibility, PHA have been developed for applications as medical implants, drug delivery matrices, and devices to support cell growth. Lots of studies showed that PHA matrices improved cell proliferation and tissue regeneration. However, the possibility of whether rapid cell proliferation on PHA matrices will induce tumor formation is unclear. Here we confirmed that proliferating rat osteoblasts grown on films of various PHA including PHB, PHBV, P3HB4HB, PHBHHx and PHBVHHx did not lead to cancer induction at least for p8th. Cell proliferation was evaluated by the incorporation of 5-bromodeoxyuridine (BrdU), the transcript expression of cancer related genes Ki67, p53 and c-Fos was monitored by quantitative Real-time PCR, the results showed the cells proliferating on the PHA films were under normal cell cycle regulation. Moreover, DNA aneuploid and telomerase activity were only detected in the positive control UMR-108 cells; compared with cells grown on films, UMR-108 cells had longer telomeres, further demonstrated the normal status of cells proliferating on the PHA films. It indicated that the above PHA family members could be used to support cell growth without indication of susceptibility to tumor induction. These results will be important for promoting the application of PHA as new members of biomaterials.

[Tissue-like cultures of rat hepatocytes in study of phase I and phase II drug metabolism].

OBJECTIVE: To investigate phase I and phase II enzyme activities in drug metabolism with tissue-like cultures of rat hepatocytes. METHODS: The gel entrapment and spheroid culture of hepatocytes were used as tissue-like cultures and the monolayer culture was used as a control. The metabolism of phenacetin and 7-hydroxycoumarin (7-HC) was evaluated as the activities of phase I and phase II enzymes after incubated in medium for a period of time. The metabolites were assayed by HPLC. The hepatocytes were exposed to beta-naphthoflavone (BNF, 50 micromol x L(-1)) before the phase I and phase II enzyme activities were analyzed. RESULT: In monolayer culture, phase I parameters decreased quickly and did not detected at d 5, and the phase II enzyme activities were not detected at d 7. In other two models of tissue-like cultures, the activities of phase I and phase II enzyme maintained at 32%-50% of the initial value at d 7. Paracetamol formation rates in spheroid culture maintained at 96% of that at d 1. The phase I enzyme activities of the spheroid culture were maintained from d 1 to d 3 at a level of 2.7-3.9-fold higher than the monolayer culture. After exposure to BNF the activities on phase I enzyme increased by about 2.5-fold (P <0.05) in all three culture models, while the increase in phase II enzyme was not significant. CONCLUSION: The gel entrapment culture and spheroid culture are superior to the monolayer culture in maintenance of drug metabolic enzyme activities.

Tissue-like cultures of rat hepatocytes in study of phase I and phase II drug metabolism / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate phase I and phase II enzyme activities in drug metabolism with tissue-like cultures of rat hepatocytes.</p><p><b>METHODS</b>The gel entrapment and spheroid culture of hepatocytes were used as tissue-like cultures and the monolayer culture was used as a control. The metabolism of phenacetin and 7-hydroxycoumarin (7-HC) was evaluated as the activities of phase I and phase II enzymes after incubated in medium for a period of time. The metabolites were assayed by HPLC. The hepatocytes were exposed to beta-naphthoflavone (BNF, 50 micromol x L(-1)) before the phase I and phase II enzyme activities were analyzed.</p><p><b>RESULT</b>In monolayer culture, phase I parameters decreased quickly and did not detected at d 5, and the phase II enzyme activities were not detected at d 7. In other two models of tissue-like cultures, the activities of phase I and phase II enzyme maintained at 32%-50% of the initial value at d 7. Paracetamol formation rates in spheroid culture maintained at 96% of that at d 1. The phase I enzyme activities of the spheroid culture were maintained from d 1 to d 3 at a level of 2.7-3.9-fold higher than the monolayer culture. After exposure to BNF the activities on phase I enzyme increased by about 2.5-fold (P <0.05) in all three culture models, while the increase in phase II enzyme was not significant.</p><p><b>CONCLUSION</b>The gel entrapment culture and spheroid culture are superior to the monolayer culture in maintenance of drug metabolic enzyme activities.</p>

[Adefovir dipivoxil in treatment of decompensated liver cirrhosis patients with YMDD mutation].

OBJECTIVE: To evaluate the efficacy and safety of adefovir dipivoxil in treatment of decompensated liver cirrhosis patients with YMDD motif mutation during lamivudine therapy. METHODS: The disease relapsed in 14 hepatitis B patients with decompensated liver cirrhosis during lamivudine treatment due to the YMDD motif mutation. All 14 patients had positive HVBDNA and active hepatitis, and were evaluated as Child-Pugh Score C (CPS-C). The patients were treated with lamivudine 50 mg/d and adefovir dipivoxil 10 mg/d for 6 months. RESULTS: One patient signed off due to non-hypoxemic hyperlactacidemia; other 13 patients showed decreased serum HBVDNA. All patients had serum HBVDNA < or =10(5) copies/ml and 7 patients had HBVDNA < or =10(4) copies/ml. Six patients regained normal serum ALT level and Child-Pugh scores decreased in all patients. CONCLUSION: Adefovir dipivoxil has satisfied efficacy and safety in treatment of decompensated liver cirrhosis patients with YMDD motif mutation during lamivudine treatment.

Adefovir dipivoxil in treatment of decompensated liver cirrhosis patients with YMDD mutation / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of adefovir dipivoxil in treatment of decompensated liver cirrhosis patients with YMDD motif mutation during lamivudine therapy.</p><p><b>METHODS</b>The disease relapsed in 14 hepatitis B patients with decompensated liver cirrhosis during lamivudine treatment due to the YMDD motif mutation. All 14 patients had positive HVBDNA and active hepatitis, and were evaluated as Child-Pugh Score C (CPS-C). The patients were treated with lamivudine 50 mg/d and adefovir dipivoxil 10 mg/d for 6 months.</p><p><b>RESULTS</b>One patient signed off due to non-hypoxemic hyperlactacidemia; other 13 patients showed decreased serum HBVDNA. All patients had serum HBVDNA < or =10(5) copies/ml and 7 patients had HBVDNA < or =10(4) copies/ml. Six patients regained normal serum ALT level and Child-Pugh scores decreased in all patients.</p><p><b>CONCLUSION</b>Adefovir dipivoxil has satisfied efficacy and safety in treatment of decompensated liver cirrhosis patients with YMDD motif mutation during lamivudine treatment.</p>

Efficacy and safety of lamivudine treatment for chronic hepatitis B in pregnancy.

AIM: To evaluate the efficacy and safety of lamivudine treatment of chronic hepatitis B disease in pregnancy. METHODS: The study group was comprised of 38 chronic HBV patients who were diagnosed pregnant during lamivudine treatment and voluntary to continue the same therapy. The control group was from documented patient data in the literatures. We compared the following parameters with those of a control group: anti-HBV efficacy, complications of pregnancy (abortion, preterm birth, neonatal asphyxia, fetal death, and congenital anomaly), incidence of HBV-positive babies and developmental anomalies in pregnant women treated with lamivudine. RESULTS: The blocking rate of lamivudine treatment was significantly higher than that of active vaccine immunization for babies with double-positive (HBsAg/HBeAg) mothers with 30-30-10 mug doses of vaccine (74.07%) and with 30-20-10 mug (64.87%). The natural vertical HBV transmission from mother to infant of "double-positive" mothers was 100% (10/10). No pregnancy complication was noted during the observation period, but in the control group the incidences of pregnancy complication were 16.67% (abortion), 43.02%(preterm), 15.62% (neonatal asphyxia), and 4.49% (fetal death), 10.0% (congenital anomaly). No HBV-positive newborn was detected and no developmental anomaly was found in the study group. CONCLUSION: Lamivudine is helpful to prevent maternal-infant HBV transmission and may reduce the complications of HBV-infected pregnant patients.

[Distribution of lamivudine- resistant variants in hepatitis B virus].

OBJECTIVE: To observe the distribution of HBV variants resistant to lamivudine and their relation to clinical manifestations of chronic hepatitis. METHODS: Using direct sequencing, YMDD (tyrosine-methionine-aspartate-aspartate) variants in patients with chronic HBV were detected before and during treatment with lamivudine. A statistical analysis of the distribution of HBV strains resistant to lamivudine was performed. RESULT: Four variant strains existed in patients before lamivudine treatment, 128 variant resistant strains were noted after 6 mouths of lamivudine treatment including 42 YVDD (valine) variants, 20 YIDD (isoleusine) variants and 66 non-YMDD variants. According to the hepatitis severity, 8 patients were mild, 108 moderate and 12 severe. Viral loading was higher and clinical types were more severe in no-YMDD variants. CONCLUSION: Variant strains including strains resistant to lamivudine exist naturally before lamivudine treatment, but lamivudine-resistant ones become more dominant after treatment. Liver inflammation is more severe in non-YMDD group.

Distribution of lamivudine- resistant variants in hepatitis B virus / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To observe the distribution of HBV variants resistant to lamivudine and their relation to clinical manifestations of chronic hepatitis.</p><p><b>METHODS</b>Using direct sequencing, YMDD (tyrosine-methionine-aspartate-aspartate) variants in patients with chronic HBV were detected before and during treatment with lamivudine. A statistical analysis of the distribution of HBV strains resistant to lamivudine was performed.</p><p><b>RESULT</b>Four variant strains existed in patients before lamivudine treatment, 128 variant resistant strains were noted after 6 mouths of lamivudine treatment including 42 YVDD (valine) variants, 20 YIDD (isoleusine) variants and 66 non-YMDD variants. According to the hepatitis severity, 8 patients were mild, 108 moderate and 12 severe. Viral loading was higher and clinical types were more severe in no-YMDD variants.</p><p><b>CONCLUSION</b>Variant strains including strains resistant to lamivudine exist naturally before lamivudine treatment, but lamivudine-resistant ones become more dominant after treatment. Liver inflammation is more severe in non-YMDD group.</p>

[Preliminary observation on the efficiency and safety of Lamivudine used in decompensative hepatic cirrhosis (B) accompanied by hypersplenism]

OBJECTIVE: To study the efficacy and safety in patients with decompensative hepatic cirrhosis treated with Lamivudine. METHODS: Eighteen decompensative hepatic cirrhosis (B) (active phage) patients accompanied with hypeersplenism were treated with Lamivudine 100mg po. per day. The total course of treatment was 3 months to 6 months when HBVDNA became negative and HBeAg seroconversion occurred in these patients after Lamivudine treatment. The efficacy and safety in patients were evaluated as follows: HBVDNA were negative, HBeAg seroconversion occurred and hepatic cirrhosis child-stageing changed. The efficacy and safety between treated group and contrast group were compared during treatment with Lamifudine for 1 year and follow-up foe 1 year after completing treatment. RESULTS: The total efficacy of treated group was 27.7% and 71.43% respectively during the phase II trial and the safety was good in these patients. CONCLUSION: The efficacy and safety of Lamivudine are good while it is used in non-registered adaptation of decompensative hepatic cirrhosis with hypersplenism.

Preliminary observation on the efficiency and safety of Lamivudine used in decompensative hepatic cirrhosis (B) accompanied by hypersplenism / 浙江大学学报·医学版

OBJECTIVE: To study the efficacy and safety in patients with decompensative hepatic cirrhosis treated with Lamivudine. METHODS: Eighteen decompensative hepatic cirrhosis (B) (active phage) patients accompanied with hypeersplenism were treated with Lamivudine 100mg po. per day. The total course of treatment was 3 months to 6 months when HBVDNA became negative and HBeAg seroconversion occurred in these patients after Lamivudine treatment. The efficacy and safety in patients were evaluated as follows: HBVDNA were negative, HBeAg seroconversion occurred and hepatic cirrhosis child-stageing changed. The efficacy and safety between treated group and contrast group were compared during treatment with Lamifudine for 1 year and follow-up foe 1 year after completing treatment. RESULTS: The total efficacy of treated group was 27.7% and 71.43% respectively during the phase II trial and the safety was good in these patients. CONCLUSION: The efficacy and safety of Lamivudine are good while it is used in non-registered adaptation of decompensative hepatic cirrhosis with hypersplenism.