[Current status of liver transplantation for adult patients with acute-on-chronic liver failure].

Acute-on-chronic liver failure(ACLF) is the most severe form of acute decompensation that develops in patients with chronic liver disease or liver cirrhosis,and is always accompanied by one or more extrahepatic organ failure, and has an extremely poor short-term prognosis. The causes triggering ACLF are complex and diverse,and the clinical stage and the type and the definition of organ failure differ greatly from one another. Therefore, a universally accepted diagnostic criteria for ACLF is not to be defined, and the epidemiological data and patient outcomes on ACLF are not easy to predict and compare among different regions. Accumulating evidence has shown that liver transplantation(LT) plays a significant role in the surgical treatment of patients with ACLF,but its clinical value is still controversial. The specific management and treatment strategy after the admission of patients with ACLF has not yet formed a unified and standardized process or opinions, which includes the monitoring in the ICU,the support and maintenance of organ functions, the selection of the surgical indication and the timing for LT and so on. Moreover, there still exists many controversies concerning, for example, whether patients with ACLF should receive greater priority for organ allocation compared to other potential candidates on the waiting list. Besides, more prospective controlled studies are urgently needed to investigate the role of the artificial liver support system in the bridging therapy to LT. The aim of this article is to review the indication selection of patients with ACLF suitable for LT,the survival outcomes and prognostic factors after LT, the selection of timing, the organ allocation policy and the bridging therapy to LT, which intends to provide new direction for designing the future clinical studies on LT in patients with ACLF.

Prognostic value of peripheral blood markers in patients with myositis-associated interstitial lung diseases.

Objectives: The aim of this study was to investigate the association between survival of anti-MDA5 autoantibody-positive/negative patients with myositis-associated interstitial lung disease (MA-ILD) and neutrophil-lymphocyte ratio (NLR), platelet-lymphocyte ratio (PLR), monocyte-lymphocyte ratio (MLR), C-reactive protein-albumin ratio (CAR), and erythrocyte sedimentation rate-albumin ratio (EAR).Method: The study included 104 patients diagnosed with MA-ILD between January 2017 and February 2019 at the First Affiliated Hospital, University of Science and Technology of China. The clinical and laboratory results were compared between survivors and non-survivors in anti-MDA5 autoantibody-positive and anti-MDA5 autoantibody-negative patients. Cox proportional hazard models were used for univariable and multivariate analyses to determine survival-related factors. A logistic regression model was used to establish a joint diagnosis, and the feasibility of the combined diagnosis to evaluate the prognosis of MA-ILD was explored.Results: Among 47 anti-MDA5-positive patients with MA-ILD, EAR was an independent predictor of survival. When separated into high and low subgroups, high MLR (> 0.604) and EAR (> 1.458) were predictive of survival (p < 0.05). High MLR, high EAR, and age combined with lactate dehydrogenase were the highest (0.886) in predicting the prognosis of MA-ILD, and were higher than the area under the curve diagnosed separately. In 57 anti-MDA5-negative patients with MA-ILD, NLR and high EAR (> 0.872) were independent predictors of survival (p < 0.05).Conclusion: MLR and EAR are associated with prognosis in anti-MDA5-positive patients. NLR and EAR are associated with prognosis in anti-MDA5-negative patients. Using NLR, MLR, and EAR, inflammatory conditions of MA-ILD can be predicted and possible outcomes estimated.

[Correlation between the changes of innate lymphoid cells in peripheral blood of systemic lupus erythematosus and its clinical significance].

Objective: To explore the function and role of innate lymphoid cells in the pathogenesis of systemic lupus erythematosus (SLE) at different disease activity levels. Methods: From Nov 2017 to May 2018, 40 patients with SLE and 15 age-matched healthy non-immune-related diseases controls were enrolled from Anhui provincial hospital. According to the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI)-2K, the patients were divided into active group (n=20) and remission group (n=20). The frequency of ILCs, B cells, CD4+T and CD8+T cells from peripheral blood mononuclear cells (PBMCs) was detected by flow cytometry. The subsets of ILCs in each group were compared with the subsets of B cells and T cell respectively. The levels of IL-4, IL-33 and IFN-γ in each group were tested by ELISA. Result: Compared with the control group, ILC1 percentage was significantly increased in SLE active group [(22.33%±2.52%) vs (14.56%±1.28%), P=0.018 1]; ILC2 percentage was decreased significantly in both remission group [(19.67%±1.83%) vs (42.48%±3.46%), P<0.000 1] and active group [(8.67%±0.83%) vs (19.67%±1.83%), P<0.000 1]; ILC3 percentage was decreased significantly in active group [(5.72%±1.08%) vs (14.35%±2.40%), P=0.001 3]. SLEDAI score was negatively correlated with the percentage of ILC2 (P=0.023 9) in all patients. The percentage of ILCs in the remission group (P=0.046 2) and activity group (P=0.003 7) were both increased significantly. Moreover, the percentage of ILC2 in active group was negatively correlated with CD4+T cells (P=0.030 8), and the serum IgG was negatively correlated with ILC2% in all patients (P=0.013 8). Compared with control group or remission group, the levels of IFN-γ (F=10.91, P=0.000 1) and IL-4 (F=6.046, P=0.004 7) in active group were remarkable higher. However, IL-33 was significantly reduced in active group (F=6.645, P=0.002 7). The percentage of ILC2 (r=0.154 3, P=0.028 8) and ILC3 (r=0.313 6, P=0.001 1) in all patients with SLE were positively correlated with the level of IL-4. Conclusion: The percentage of ILCs is related to disease activity, and ILCs play a "double-edged" role in the pathogenesis of SLE. Its function and mechanism are worth further exploration.

Enhanced efficacy in anti-tumour activity by combined therapy of recombinant FGFR-1 related angiogenesis and low-dose cytotoxic agent.

Fibroblast growth factor receptor-1 (FGFR-1) has been used as a target for anti-angiogeneic therapy of cancer. The strategies of combining anti-angiogenic biotherapy with chemotherapeutic drugs show potential and promise for cancer therapy. In this study, we evaluated the anti-tumour efficacy of chicken FGFR-1 (cFR-1) vaccine combined with low-dose gemcitabine in two mice tumour models. We found that both the cFR-1 vaccine and low-dose gemcitabine can suppress tumour growth to some extent. Remarkably, the combination strategy produces an apparent decrease in tumour volume, microvessel density and tumour cell proliferation, and an increase of apoptosis without obvious side-effects compared with either therapy alone. Moreover, the combination strategy also demonstrated synergistic indices against tumour growth and angiogenesis. Furthermore, auto-antibodies against mouse FGFR-1 were identified. These findings support the idea that the combination strategy synergistically strengthens anti-tumour activity via suppression of tumour angiogenesis without overt toxicity in tumour-bearing mice.

Regulation of K3 keratin gene transcription by Sp1 and AP-2 in differentiating rabbit corneal epithelial cells.

Rabbit corneal epithelial cells cultured in the presence of 3T3 feeder cells undergo biochemical differentiation, as evidenced by their initial expression of K5 and K14 keratins characteristic of basal keratinocytes, followed by the subsequent expression of K3 and K12 keratin markers of corneal epithelial differentiation. Previous data established that mutations of an Sp1 site in a DNA element, E, that contains overlapping Sp1 and AP-2 motifs reduce K3 gene promoter activity by 70% in transfection assays. We show here that Sp1 activates while AP-2 represses the K3 promoter. Although undifferentiated corneal epithelial basal cells express equal amounts of Sp1 and AP-2 DNA-binding activities, the differentiated cells down-regulate their Sp1 activity slightly but their AP-2 activity drastically, thus resulting in a six- to sevenfold increase in the Sp1/AP-2 ratio. This change coincides with the activation and suppression of the differentiation-related K3 gene and the basal cell-related K14 keratin gene, respectively. In addition, we show that polyamines, which are present in a high concentration in proliferating basal keratinocytes, can inhibit the binding of Sp1 to its cognate binding motif but not that of AP-2. These results suggest that the relatively low Sp1/AP-2 ratio as well as the polyamine-mediated inhibition of Sp1 binding to the E motif may account, in part, for the suppression of the K3 gene in corneal epithelial basal cells, while the elevated Sp1/AP-2 ratio may be involved in activating the K3 gene in differentiated corneal epithelial cells. Coupled with the previous demonstration that AP-2 activates the K14 gene in basal cells, the switch of the Sp1/AP-2 ratio during corneal epithelial differentiation may play a role in the reciprocal expression of the K3 and K14 genes in the basal and suprabasal cell layers.

Specific tumor immunity induced with mitomycin C-treated syngeneic tumor cells (MCT). Effects of carrageenan and trypan blue on MCT-induced immunity in mice.

Mitomycin C treatment of 3-methylcholanthrene-induced syngeneic tumor cells in male mice of the highly inbred strain CBA/WEHI was a convenient and effective method for producing viable but nondividing tumor cells. A single sc injection of 1 x 10(6) mitomycin C-treated syngeneic tumor cells (MCT) induced a specific antitumor immune response that could mediate rejection of small subcutaneous tumor grafts and substantially inhibit growth of larger grafts in normal mice. The response was evident between 3 and 7 days after immunization and persisted for at least 120 days. Adoptive transfer experiments with peritoneal exudate cells from MCT-immunized mice indicated that a cellular basis for resistance was likely in the absence of a detectable humoral component. Administration of MCT to mice with established tumor grafts did not, however, affect tumor growth. Experiments with carrageenan and trypan blue showed that neither induction nor expression of cytotoxic effector cells in MCT-immunized mice was inhibited by these substances, though carrageenan and trypan blue augmented tumor growth in normal mice.

Uroplakin II gene is expressed in transitional cell carcinoma but not in bilharzial bladder squamous cell carcinoma: alternative pathways of bladder epithelial differentiation and tumor formation.

Uroplakins (UPs) are integral membrane proteins that are synthesized as the major differentiation products of mammalian urothelium. We have cloned the human UP-II gene and localized it on chromosome 11q23. A survey of 50 transitional cell carcinomas (TCCs) revealed a UP-II polymorphism but no tumor-specific mutations. Immunohistochemical staining using rabbit antisera against a synthetic peptide of UP-II and against total UPs showed UP reactivity in 39.5% (17 of 43 cases) of conventional TCCs, 12.8% (5 of 39) of bilharzial-related TCCs, and 2.7% (1 of 36) of bilharzial-related squamous cell carcinomas (SCCs). The finding that fewer bilharzial TCCs express UPs than conventional TCCs (12.8 versus 40%) raised the possibility that the former are heterogeneous, expressing SCC features to varying degrees. Our data strongly support the hypothesis that urothelium can undergo at least three pathways of differentiation: (a) urothelium-type pathway; (b) epidermis-type pathway; and (c) glandular-type pathway, characterized by the production of UPs, K1/K10 keratins, and secreted glycoproteins, respectively. Vitamin A deficiency and mesenchymal factors may play a role in determining the relative contributions of these pathways to urothelial differentiation as well as to the formation of TCC, SCC, and adenocarcinoma, or a mixture thereof.

Modulation of a slowly inactivating potassium current, I(D), by metabotropic glutamate receptor activation in cultured hippocampal pyramidal neurons.

I(D) is a slowly inactivating 4-aminopyridine (4-AP)-sensitive potassium current of hippocampal pyramidal neurons and other CNS neurons. Although I(D) exerts multifaceted influence on CNS excitability, whether I(D) is subject to modulation by neurotransmitters or neurohormones has not been clear. We report here that one prominent effect of metabotropic glutamate receptor (mGluR) activation by short (3 min) exposure to 1S, 3R-1-aminocyclopentane-1,3-dicarboxylic acid (1S,3R-ACPD) (100 microM) is suppression of I(D) by acceleration of its inactivation. I(D) was identified as a target of mGluR-mediated modulation because inactivation of a component of outward current sensitive to 100-200 microM 4-AP was accelerated by 1S,3R-ACPD, and because 4-AP occluded any further actions of 1S,3R-ACPD. Enhancement of I(D) inactivation was induced by the group I-preferring agonist RS-3, 5-dihydroxyphenylglycine (3,5-DHPG) and the group II-preferring agonist 2S,2'R,3'R)-2-(2',3'dicarboxycyclopropyl)-glycine (DCG-IV), but not by the group III-preferring agonist L(+)-2-amino-4-phosphonobutyric acid (L-AP4); it was blocked by the broadly acting mGluR antagonist S-alpha-methyl-4-carboxyphenylglycine (S-MCPG). Furthermore, inactivation of I(D) was enhanced by inclusion of GTPgammaS in the internal solution and blocked by inclusion of GDPbetaS. Metabotropic GluR-induced suppression of I(D) was manifest in three aspects of excitability previously linked to I(D) by their sensitivity to 4-AP: reduction in input conductance and enhanced excitability at voltages just positive to the resting potential, reduced delay to action potential firing during depolarizing current injections, and delayed action potential repolarization. We suggest that mGluR-induced suppression of I(D) could contribute to enhancement of hippocampal neuron excitability and synaptic connections.

Mapping quantitative trait loci by genotyping haploid tissues.

Mapping strategies based on a half- or full-sib family design have been developed to map quantitative trait loci (QTL) for outcrossing species. However, these strategies are dependent on controlled crosses where marker-allelic frequency and linkage disequilibrium between the marker and QTL may limit their application. In this article, a maximum-likelihood method is developed to map QTL segregating in an open-pollinated progeny population using dominant markers derived from haploid tissues from single meiotic events. Results from the haploid-based mapping strategy are not influenced by the allelic frequencies of markers and their linkage disequilibria with QTL, because the probabilities of QTL genotypes conditional on marker genotypes of haploid tissues are independent of these population parameters. Parameter estimation and hypothesis testing are implemented via expectation/conditional maximization algorithm. Parameters estimated include the additive effect, the dominant effect, the population mean, the chromosomal location of the QTL in the interval, and the residual variance within the QTL genotypes, plus two population parameters, outcrossing rate and QTL-allelic frequency. Simulation experiments show that the accuracy and power of parameter estimates are affected by the magnitude of QTL effects, heritability levels of a trait, and sample sizes used. The application and limitation of the method are discussed.

Isolation and characterization of a mammalian homolog of the Drosophila white gene.

The Drosophila melanogaster white gene is a member of the ABC transporter superfamily of ATPase transmembrane proteins and is involved in the cellular uptake of guanine and tryptophan. We have cloned and sequenced human and mouse homologs of white which share 55-58% amino acid similarity with the Drosophila protein. Northern analysis reveals that the mammalian homolog is highly expressed in several tissues, including brain, spleen, lung and placenta. We have localized the gene to human chromosome 21q22.3 by means of fluorescence in situ hybridization and linkage analysis using a (CA)n polymorphism. The human homolog maps to the interval between D21S212 and D21S171, a region which includes loci for bipolar affective disorder and a recessive form of deafness. Since tryptophan is a precursor for the neurotransmitter serotonin and neurotoxic metabolites of the kynurenine pathway, we propose that the human homolog of white is a suitable candidate gene for these neurological disorders in humans.

In vitro growth and differentiation of rabbit bulbar, fornix, and palpebral conjunctival epithelia. Implications on conjunctival epithelial transdifferentiation and stem cells.

PURPOSE: The anterior surface of the eye is covered by several physically contiguous but histologically distinguishable epithelial overlying the cornea, limbus, bulbar conjunctiva, fornix conjunctiva, and palpebral conjunctiva. It is important to determine whether the different phenotypes of these epithelia are the result of intrinsic divergence, extrinsic modulation, or a combination of both. Based on keratin expression and cell kinetic criteria, the authors previously suggested that corneal epithelial stem cells may actually reside in the limbal basal layer. METHODS: In this article, the relationship between the corneal-limbal epithelial cells and conjunctival epithelial cells was analyzed by comparing their growth and differentiation properties in an identical cell culture environment. RESULTS: Using Dispase instead of trypsin to dissociate the cells, the authors were able to grow all five rabbit ocular surface epithelia in the presence of 3T3 feeder cells. They found that corneal and limbal cells synthesize identical keratins, including large amounts of the K3 and K12 markers of corneal-type differentiation. By contrast, all three conjunctival epithelia shared another keratin pattern, with large amounts of simple epithelial keratins but only minute amounts of K3/K12 keratins. CONCLUSIONS: This observation, coupled with previous findings that the "transdifferentiation" of conjunctival epithelial cells to corneal epithelium appears to be both incomplete and reversible, provides strong evidence that (1) the limbal-corneal epithelial cells form a lineage distinct from the conjunctival lineage and (2) conjunctival transdifferentiation actually represents a process of environmental modulation. In addition, of the three types of conjunctival epithelial cells, fornix cells were found to have a much greater proliferative potential than bulbar and palpebral cells. This observation, coupled with recent finding that fornix is enriched in slow-cycling (label-retaining) cells, raises the possibility that conjunctival epithelial stem cells may preferentially reside in the fornix.

Age-related changes in the synaptic plasticity of rat superior cervical ganglia.

A few seconds of tetanic preganglionic stimulation of rat sympathetic ganglia results in potentiation of cholinergic synaptic transmission lasting several hours. However, ganglia from aged (28-32 months) rats did not develop as much potentiation as did ganglia from young (3-6 months) rats. This potentiation appears to decay exponentially but in two phases. The early component decays in 15 minutes, a time course consistent with the phenomenon of post-tetanic potentiation (PTP). The later component decays over several hours and is a form of long-term potentiation (LTP). A double exponential decay model was employed to quantitatively resolve the decay of potentiation and allow quantitative comparison of the decay parameters in both aged and young rats. It is clear that the magnitude and duration of PTP was the same in both age groups. However both the magnitude and decay time-constant for LTP were 30-50% smaller in the aged group of rats. Several agents which mimic cyclic AMP or stimulate cyclic AMP production in the ganglion enhance nicotinic transmission for several hours. However, these agents, 8-bromo cAMP, forskolin, isoproterenol, and secretin were equally effective in potentiating transmission in ganglia from both young and aged rats. Previous studies demonstrated that stimulus induced LTP and cyclic AMP induced potentiation was the result of enhanced release of acetylcholine (ACh) and not from increased postsynaptic responsiveness. Presumably these agents act through the same pathway and by the same mechanism that generates LTP following preganglionic tetany. These observations demonstrate that there is a selective age-dependent decline in the capacity for sympathetic ganglia to generate long-term changes in synaptic efficacy.(ABSTRACT TRUNCATED AT 250 WORDS)

The profiles of human and primate [3H]Nalpha-methylhistamine binding differ from that of rodents.

Characterization of the histamine H3 receptor in rodent species has been extensive but limited characterization has been done with primate or human tissue. We have characterized the binding of [3H]Nalpha-methylhistamine to cynomolgus monkey and human brain membranes to determine whether there are any significant differences among species' pharmacology. In monkey, [3H]Nalpha-methylhistamine bound, in a guanine nucleotide-sensitive fashion, to an apparently homogeneous class of sites at equilibrium (K(D) = 1.4 nM, Bmax = 34 fmol/mg protein). The profile of binding was broadly similar to that of rodents, with a couple of significant differences. Most notably, the potency of the histamine H3-receptor-specific antagonist thioperamide (Ki = 240 nM) was substantially less than reported for rodents and under assay conditions that yield a two-site curve fit in rodents only a single class of thioperamide binding sites was detected in monkey. Burimamide, however, yielded a two-site curve fit (KiH = 6.7 nM, KiL = 1100 nM) independent of the presence of sodium in the assay, as it does in rodents. Characterization of the human brain histamine H3 receptor showed that it was similar to the monkey and not rodent receptor. Our findings indicate that differences between primate and rodent histamine H3 receptors of potentially serious importance for the discovery of antagonists active in humans do exist.

alpha-Tocopherol in the developing rat retina: a high pressure liquid chromatographic analysis.

High pressure liquid chromatography was used to measure alpha-tocopherol in the retinas of rats reared in a cyclic light or dark environment. These measurements were performed on extracts of whole retinas during the developmental period, 18-60 days, and on isolated ROS from adult animals. Similar alpha-tocopherol determinations were performed on retinas and isolated ROS following exposure of rats to intense visible light for 24 hr periods. The results show that alpha-tocopherol is chromatographically separated from the vitamin A derivatives found in the retina and is pure, as judged by mass spectrometry. In the retinas of cyclic light and dark reared rats, alpha-tocopherol accumulates in an age dependent fashion, so that at 60 days the level is nearly double that of animals at 18-20 days of age (P less than 0.001). Because the age dependent accumulation of rhodopsin is greater in dark reared rats, the average molar ratio of rhodopsin to alpha- tocopherol in the retina of dark reared animals is 25% higher than in cyclic light rats. Following exposure of rats to intense visible light for 24 hr periods, alpha-tocopherol concentrations in the retina were unchanged from the levels in control animals. In adult animals the concentration of alpha-tocopherol in ROS is 2.5-3.5 times higher than in whole retina. ROS from adult cyclic light reared rats also contain an average of 43% more alpha-tocopherol per mg protein than ROS from dark maintained animals (P less than 0.005).(ABSTRACT TRUNCATED AT 250 WORDS)

Expression of K12 keratin in alkali-burned rabbit corneas.

The healing of alkali-injured corneas is characterized by the persistence of polymorphonuclear leukocytes (PMN) in tissues and recurrent corneal epithelial defects. It has been suggested that the proteolytic enzymes secreted by PMN may account in part for the recurrent epithelial defects in the alkali-burned corneas. Cytoplasmic keratins, which form intracellular intermediate filaments, participate in the formation of hemidesmosomes and play a key role in the focal adhesion of epithelial cells to the basement membranes. The K3/K12 keratin pair is a major constituent of differentiated and stratified corneal epithelium. We have recently cloned the cDNA encoding the rabbit K12 keratin. In the present study we examined the expression of K12 keratin during the healing of alkali-burned rabbit corneas by slot-blot and in situ hybridization. Our results indicate that in normal cornea K12 keratin is equally expressed in all cell layers of stratified corneal epithelium and suprabasal layers of limbal epithelium, but not in bulbar conjunctival and other epithelia, i.e., lens, iris, and retinal pigment epithelium. The basal cells of the detached regenerating epithelium of the injured cornea express a very low level of K12 keratin. These observations are consistent with the notion that defective expression of K3/K12 keratins may play a role in the abnormal attachment of the regenerating epithelium to the basement membrane.

Diamond-like carbon as biological compatible material for cell culture and medical application.

Ion beam assisted diamond-like carbon (DLC) films have been used for growing the human hematopoietic myeloblastic ML-1 cells and human embryo kidney 293 cells in the control environment. DLC films were directly deposited onto the P-35 plastic dishes by impacting the high kinetic energy (1000 eV) of methane ions at room temperature. The present results showed that both ML-1 and HEK 293 cells continuously grow with and without DLC films. It has demonstrated that human cells proliferated on DLC film with very high viability and DLC material had no toxicity to cultured human ML-1 and HEK 293 cells. We conclude that DLC film is a biological compatible material for potential cell culture matrix and bio-medical applications.

[Significance of ultrastructure of peripheral nerve tumors in pathological and differential diagnoses].

Biopsy material fixed with formalin in 10 cases of peripheral nerve tumors and 5 soft tissue tumors were studied with special reference to the significance of the ultrastructure of peripheral nerve tumors in histopathological and differential diagnosis. The results demonstrated that electron microscopic examination of formalin-fixed tissue may still reveal their ultrastructural characteristics. This is of certain practical value in pathological and differential diagnosis in some difficult peripheral nerve tumors.

[Morphology, diagnosis and differential diagnosis of mesothelioma].

Forty cases of mesothelioma biopsied and autopsied from 1961 to 1985 were studied and analysed as to their morphology, diagnosis and differential diagnosis. Mesothelioma had complex histologic types. The tumor cells varied in size and shape and were characterized morphologically by the coexistence of biphasic differentiation, adenoid and microvilli formation. The pathologic diagnosis is made according to the feature of biphasic differentiation, or coexistence of epithelioid and spindle tumor cells. It should be emphasized that the tissue sections are taken from multiple areas. Usually, the correct diagnosis is made in correlation to the clinical manifestations. For difficult cases of the diffuse mesothelioma, electron microscopy is needed to differentiate from metastatic adenocarcinoma and spindle cell sarcoma etc.