[Evaluation of ocular surface status and function in primary Sjögren's syndrome with hypothyroidism].

OBJECTIVE: To explore the effect of hypothyroidism (HT) on the ocular surface status of patients with primary Sjögren's syndrome-related dry eye (pSS-DED). METHODS: The cross-sectional study included 36 patients with pSS-DED who were treated at the dry eye clinic of Peking University Third Hospital from December 2020 to June 2021, of whom 12 were pSS-DED patients combined with HT. In the same period, 24 patients with simple dry eye disease (DED) were served as a control group. All the patients filled out the Ocular Surface Disease Index (OSDI) questionnaire, and performed tear film break-up time (BUT), Schirmer test, tear meniscus height, corneal/conjunctival fluorescein staining, meibomian gland secretion capacity, meibum evaluation and confocal microscope examination. RESULTS: (1) Compared with pSS-DED and simple DED patients, pSS-DED +HT patients had lower average BUT [(2.7±0.8) s], Schirmer test [(4.9±4.8) mm] and tear meniscus height [(0.13±0.03) mm], and the difference was statistically significant (F=12.43, P < 0.01; F=6.96, P < 0.01; F=3.31, P < 0.05). (2) Compared with DED and pSS-DED patients, the meibomian gland secretion capacity and meibomian trait scores of pSS-DED+HT patients were mainly distributed in the high division. There were statistically significant differences in the distribution of secretion capacity of meibomian glands (χ2=10.72, P < 0.05) and meibomian trait assessment scores (χ2=8.34, P < 0.05) among the three groups. (3) Serum total thyroxine and serum free thyroxine levels in the pSS-DED+HT patients showed positive correlation (P < 0.05, P < 0.05) with their BUT (r=0.60, 0.60), Schirmer's test (r=0.64, 0.66) and tear river height (r=0.61, 0.62), independent of lid gland secretory capacity; no significant correlation was found between thyroid-stimulating hormone, anti-thyroglobulin antibody and lid gland secretory capacity. Thyroid hormone, anti-thyroglobulin antibody, and thyroid peroxidase antibody were not found to be significantly correlated with ocular surface status. (4) Compared with pSS-DED, the fiber density of the subbasal nerve plexus in pSS-DED+HT group decreased (t=2.06, P < 0.05), and the curvature score increased (t=2.13, P < 0.05). CONCLUSION: The ocular surface condition of pSS-DED patients with HT is worse than that of pSS-DED and DED patients. The main manifestations are that tear secretion, tear film stability, secretory function of the meibomian glands, meibum trait and fiber density of the subbasal nerve plexus decrease while the curvature increases. The mechanism might be related to the decrease in thyroid hormone production.

[Progress in the clinical diagnosis and treatment of hepatic vascular diseases].

The liver has a very special dual blood supply, including the portal vein (65%~75%) and hepatic artery (25%~35%). The hepatic veins returns blood to the systemic circulation via the portal vein, and hepatic artery after hepatic sinusoidal confluence. The lesions on the hepatic vein and its branches can cause ischemia and hypoxia or obstruction of the drainage system, portal hypertension, upper gastrointestinal variceal bleeding, hepatic encephalopathy, and so on. Clinically, hepatic vascular diseases are relatively rare, so the diagnosis and treatment are relatively difficult. Herein, we review the diseases related to the hepatic vascular system.

[Value of serum alpha-fetoprotein for the prognostic evaluation of hepatitis B virus-related acute-on-chronic liver failure treated with artificial liver].

Objective: To investigate the value of alpha-fetoprotein (AFP) level on survived hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF) patients treated with artificial liver. Methods: Clinical indicators of HBV-ACLF patients who were previously treated with plasma exchange-based artificial liver at our department were retrospectively collected. The difference of serum AFP level between the survival and the death group was compared at 30, 90 and 180 days after artificial liver treatment. The ROC curves of the subjects were plotted, and the sensitivity and specificity of AFP for the survival prediction of the patients at 30, 90 and 180 days after artificial liver surgery were calculated. AFP was divided into a high AFP group and a low AFP group using median value. AFP and postoperative survival predictive value at 30, 90, and 180 days were analyzed. Results: A total of 93 cases were included in this study. The AFP of the survival group at 30, 90, and 180 days was (231.0 ± 286.2) ng / ml, (237.69 ± 297) ng / ml, (229.44 ± 286.46) ng/ml, and the death group was (76.4 ± 104.7) ng/ml, (103.13 ± 116.99) ng / ml, (136.34 ± 2.9.29) ng/ml, respectively. AFP of the death group was significantly lower than the corresponding survival group (P < 0.05). Receiver operating characteristic (ROC) curve analyses indicated that the area under the curve (AUC) and its 95% confidence interval at 30, 90, and 180 days after artificial liver surgery were 0.739 (0.611 ~ 0.867), 0.675 (0.550 ~ 0.80), 0.653 (0.524 ~ 0.781), respectively. The median serum AFP value was 110 ng/ml, and the survival analysis showed that the survival time of the high AFP group was significantly higher than the low AFP group at 30 d (P = 0.01), 90 d (P = 0.04) and 180 d (P = 0.03) after artificial liver surgery. Conclusion: Serum AFP can be used as a predictor of survival for HBV-ACLF patients after artificial liver therapy and its clinical value needs to be further verified by the larger sample size.

[Umbilical cord mesenchymal stem cells and their association with liver fibrosis].

At present, the most effective therapeutic method for end-stage liver fibrosis is liver transplantation. However, the application of liver transplantation is limited by a shortage of liver donors, a high incidence rate of surgical complications, graft-versus-host disease, and high medical costs. Umbilical cord mesenchymal stem cell (UC-MSC) transplantation may become a promising method for the treatment of liver diseases. UC-MSCs are adult stem cells which exhibit multipotential differentiation and can differentiate into hepatic parenchymal cells. Due to their functions including immune regulation and secretion of trophic factors, UC-MSCs can inhibit immune response, promote hepatocyte regeneration, alleviate the progression of liver fibrosis, and improve liver function. In addition, compared with bone marrow mesenchymal stem cells, UC-MSCs have abundant sources, noninvasive collection, and high safety and thus they are attracting more and more attention. This article reviews the characteristics of UC-MSCs and their mechanism of action in the treatment of liver fibrosis, as well as risks of UC-MSCs therapy.

Clinical therapeutic effects of human umbilical cord-derived mesenchymal stem cells transplantation in the treatment of end-stage liver disease.

We aimed to evaluate clinical therapeutic effects of human umbilical cord-derived mesenchymal stem cell (UCMSC) transplantation in the treatment of end-stage liver diseases. The human UCMSCs were cultured and prepared, and then transplanted into the hepatic tissues of 50 patients with decompensated cirrhosis. The liver function, thrombin function, Model for End-Stage Liver Disease (MELD) score, and hemodynamic index value were detected during a 24-week follow-up period, with the addition of hepatoprotective, antiviral, and other conventional treatments. No complications or serious side effects were observed. In the first 2-3 weeks after surgery, symptoms including abdominal distension, oliguria, edema, and others decreased significantly, with increased appetite compared with before surgery. In the 24-week follow-up period, the levels of serum albumin and prealbumin increased significantly compared with the preoperative levels; the decrease of coagulation indicators was not significant. The MELD scores were also markedly increased. Alpha-fetoprotein levels increased without significance after treatment. There was no significant difference in the hemodynamic changes in the portal and splenic veins according to ultrasound. Moreover, no significant differences in the liver and thrombin functions between the hepatitis B virus group and the other-etiology group were observed.

Intracellular ASIC1a regulates mitochondrial permeability transition-dependent neuronal death.

Acid-sensing ion channel 1a (ASIC1a) is the key proton receptor in nervous systems, mediating acidosis-induced neuronal injury in many neurological disorders, such as ischemic stroke. Up to now, functional ASIC1a has been found exclusively on the plasma membrane. Here, we show that ASIC1a proteins are also present in mitochondria of mouse cortical neurons where they are physically associated with adenine nucleotide translocase. Moreover, purified mitochondria from ASIC1a(-/-) mice exhibit significantly enhanced Ca(2+) retention capacity and accelerated Ca(2+) uptake rate. When challenged with hydrogen peroxide (H2O2), ASIC1a(-/-) neurons are resistant to cytochrome c release and inner mitochondrial membrane depolarization, suggesting an impairment of mitochondrial permeability transition (MPT) due to ASIC1a deletion. Consistently, H2O2-induced neuronal death, which is MPT dependent, is reduced in ASIC1a(-/-) neurons. Additionally, significant increases in mitochondrial size and oxidative stress levels are detected in ASIC1a(-/-) mouse brain, which also displays marked changes (>2-fold) in the expression of mitochondrial proteins closely related to reactive oxygen species signal pathways, as revealed by two-dimensional difference gel electrophoresis followed by mass spectrometry analysis. Our data suggest that mitochondrial ASIC1a may serve as an important regulator of MPT pores, which contributes to oxidative neuronal cell death.

Mood stabilizers regulate cytoprotective and mRNA-binding proteins in the brain: long-term effects on cell survival and transcript stability.

Manic depressive illness (MDI) is a common, severe, chronic and often life-threatening illness. Despite well-established genetic diatheses and extensive research, the biochemical abnormalities underlying the predisposition to, and the pathophysiology of, these disorders remain to be clearly established. Despite formidable obstacles in our attempts to understand the underlying neurobiology of this illness, there is currently considerable excitement about the progress that is being made using novel strategies to identify changes in gene expression that may have therapeutic relevance in the long-term treatment of MDI. In this paper, we describe our recent research endeavours utilizing newer technologies, including a concerted series of mRNA RT-PCR studies, which has led to the identification of novel, hitherto completely unexpected targets for the long-term actions of mood stabilizers - the major cytoprotective protein bcl-2, a human mRNA binding (and stabilizing) protein, AUH, and a Rho kinase. These results add to the growing body of data suggesting that mood stabilizers may bring about some of their long-term benefits by enhancing neuroplasticity and cellular resilience. These results are noteworthy since recent morphometric brain imaging and post-mortem studies have demonstrated that MDI is associated with the atrophy and/or loss of neurons and glia. The development of novel treatments which more directly target molecules involved in critical CNS cell survival and cell death pathways have the potential to enhance neuroplasticity and cellular resilience, and thereby modulate the long-term course and trajectory of these devastating illnesses.

Phosphatidylinositol 4,5-bisphosphate and intracellular pH regulate the ROMK1 potassium channel via separate but interrelated mechanisms.

ROMK channels are responsible for K(+) secretion in kidney. The activity of ROMK is regulated by intracellular pH (pH(i)) with acidification causing channel closure (effective pK(a) approximately 6.9). Recently, we and others reported that a direct interaction of the channels with phosphatidyl-4,5-bisphosphate (PIP(2)) is critical for opening of the inwardly rectifying K(+) channels. Here, we investigate the relationship between the mechanisms for regulation of ROMK by PIP(2) and by pH(i). We find that disruption of PIP(2)-ROMK1 interaction not only decreases single-channel open probability (P(o)) but gives rise to a ROMK1 subconductance state. This state has an increased sensitivity to intracellular protons (effective pK(a) shifted to pH approximately 7.8), such that the subconductance channels are relatively quiescent at physiological pH(i). Open probability for the subconductance channels can then be increased by intracellular alkalinization to supra-physiological pH. This increase in P(o) for the subconductance channels by alkalinization is not associated with an increase in PIP(2)-channel interaction. Thus, direct interaction with PIP(2) is critical for ROMK1 to open at full conductance. Disruption of this interaction increases pH(i) sensitivity for the channels via emergence of the subconductance state. The control of open probability of ROMK1 by pH(i) occurs via a mechanism distinct from the regulation by PIP(2).

The mood-stabilizing agents lithium and valproate robustly increase the levels of the neuroprotective protein bcl-2 in the CNS.

Differential display of mRNA was used to identify concordant changes in gene expression induced by two mood-stabilizing agents, lithium and valproate (VPA). Both treatments, on chronic administration, increased mRNA levels of the transcription factor polyomavirus enhancer-binding protein (PEBP) 2beta in frontal cortex (FCx). Both treatments also increased the DNA binding activity of PEBP2 alphabeta and robustly increased the levels of bcl-2 (known to be transcriptionally regulated by PEBP2) in FCx. Immunohistochemical studies revealed a marked increase in the number of bcl-2-immunoreactive cells in layers 2 and 3 of FCx. These novel findings represent the first report of medication-induced increases in CNS bcl-2 levels and may have implications not only for mood disorders, but also for long-term treatment of various neurodegenerative disorders.

[Detection of hepatitis C virus RNA in cells with hepatocellular carcinoma and its practical significance by in situ polymerase chain reaction].

Persistent infection with hepatitis C virus (HCV) is associated with chronic hepatitis and cirrhosis which may eventually develop into hepatocellular carcinoma (HCC). As far, the pathogenesis mechanism of HCC is unclear and nothing is known of the distribution, frequency or type of infected cells in HCC. One-step reverse transcription in situ polymerase chain reaction (ORT-PCRIS) to detect HCV RNA in HCC samples was developed in our laboratory. Liver tissues from 27 patients with HCC were investigated by this technique for frozen or paraffin-embedded sections. Meanwhile, HCV RNA in sera and extracts of specimens with HCC were assayed by RT-PCR. The positive rate of HCV RNA by ORT-PCRIS was 81.5% (22/27) in the peritumor liver tissues and 77.8% (21/27) in the tumor tissues, significantly higher than 29.6% (8/27) in sera and 37.0% (10/27) in the extreats of HCC by RP-PCR (P < 0.01). HCV RNA positive signals were found mainly in the nuclei of tumor cell, and mainly both the nuclei and cytoplasms in peritumor cells and mainly both the nuclei and cytoplasms in peritumor cells (P < 0.05). Positive granules of HCV RNA were much more in peritumor cells than in the tumor cells. Positive cells were scattered mainly on the point-type in the cancer tissues and on the diffusion-type in the peritumor tissues. Our findings suggest that ORT-PCRIS for detecting HCV RNA in the cell with HCC remarkably prior to traditional RT-PCR. HCV infection plays a relatively important role in determination of HCC development and perhaps HCV replication and its genomic RNA integration with hepatocyte DNA are involved in the course of the malignant transformation of hepatocytes.

Effects of tetrandrine on free intracellular Ca2+ in isolated rat brain cells.

Using Ca(2+)-sensitive fluorescent indicator, Fura-2/AM, intracellular free Ca2+ ([Ca2+]i) was measured. Resting [Ca2+]i was 221 +/- 18 nmol.L-1 in the presence of Ca2+ 1.3 mmol.L-1 in Hank's solution. Tetrandrine (Tet) 30 mumol.L-1 had no effect on the resting [Ca2+]i when the extracellular Ca2+ were 0-2 mmol.L-1. In the presence of extracellular Ca2+ 1.3 mmol.L-1, Tet (1-100 mumol.L-1) concentration-dependently inhibited the high extracellular K(+)-induced [Ca2+]i elevation, with an IC50 value of 8.2 mumol.L-1 (95% confidence limits: 1.9-32.9 mumol.L-1). Low concentrations of Tet (1-10 mumol.L-1) did not alter the norepinephrine-induced [Ca2+]i elevation. Tet 30 mumol.L-1 depressed norepinephrine 10 mumol.L-1 induced [Ca2+]i elevation by 42%. The results suggested that Tet inhibited the Ca2+ influx through voltage-dependent ionic channels and, at high concentrations, through receptor-operated ionic channels in the brain cells.

[Clinical evaluation of HBV, HCV and HDV serum markers in patients with liver cirrhosis (LC) and hepatocellular carcinoma (HCC)].

11 kinds of HBV, HCV and HDV serum markers were investigated in 46 patients with HCC, 48 patients with LC, and 52 controls without liver disease. The prevalence of one out of HBV or HCV or HDV markers (M) in HCC and LC was 91.3% and 95.8% respectively, significantly higher than that in controls (17.3%). Positivity of HBV-M in HCC and LC was remarkably higher than those of HCV-M and HDV-M (P < 0.05). Prevalence of HCV-M in HBV-M negative HCC and LC was 66.7% and 75.0% respectively, significantly higher than that in HBV-M positive cases (P < 0.05). Frequency of viral replication in HCC and LC was significantly higher than that in controls (P < 0.01). The co-occurrence of two or three kinds of viral markers in HCC was more prevalent than that in LC (38% vs 14%, P < 0.05). Patients with coinfection from both HBV and HDV had a significantly younger age than those infected by HBV alone or infected by HCV (more than 10 years earlier). Among HCC and LC, 36% of HBV seronegative cases had HBV DNA detectable in their serum. Our data suggest that HCC and LC have a close association with the infection of HBV HCV and HDV especially HBV. Active viral replication and coinfection of several kinds of virus play on important role in the determination of HCC or LC development, and HDV appears to provide an additional risk for HCC and LC. In HBV-M negative cases, HCV infection may be more important for HCC and LC development than HBV.

Effects of tetrandrine on action potentials and afterhyperpolarization potentials in toad dorsal root ganglia.

Intracellular recordings were obtained from the cytosome of type A primary afferents in the isolated toad dorsal root ganglia (DRG) preparations. Bath application of Ca2+ 8.0 mmol.L-1 led to prolong action potential duration of repolarization 100% (APD100) by 23%, afterhyperpolarization potential duration of depolarization 50% (AHPD50) by 46%, and increase afterhyperpolarization potential amplitude (AHPA) by 40%. Verapamil 3 mumol.L-1 shortened the APD100 by 7% and the AHPD50 by 13%, and reduced the AHPA by 17%. The effects of Tet 3-100 mumol.L-1 consisted of APD and AHPD50 shortening and AHPA reduction in concentration-dependent manner. Tet 100 mumol.L-1, APD100 was shortened by 16%, AHPD50 by 18%, and AHPA was reduced by 20%. The results suggested that the effects of Tet may be related to its Ca2+ channel blockade in DRG.

Evolution of rhythms during periodic stimulation of embryonic chick heart cell aggregates.

During periodic stimulation of spontaneously beating chick heart cell aggregates, there is often an evolution of coupling patterns between the stimulator and the aggregate action potential. For example, at rapid stimulation frequencies, a rhythm that is initially 1:1 (stimulus frequency:aggregate frequency) can evolve to other rhythms such as 5:4 and 4:3. Time-dependent effects generated during periodic stimulation are characterized by three types of experiments to determine 1) the effect of periodic stimulation on the intrinsic cardiac beat rate (overdrive suppression), 2) the effect of periodic stimulation on the phase resetting properties of the aggregate, and 3) the time-dependent changes in the coupling patterns between the stimulator and the aggregate during periodic stimulation. The protocols involved variations in the duration and rate of periodic stimulation. A mathematical model is developed in the form of a two-dimensional finite difference equation based on the data from experiments 1 and 2. The model is used to predict the data generated by experiment 3. There is good correspondence with the experiments in that the theory reproduces complex transitions between various rhythms and displays irregular rhythms similar to those observed experimentally. These results have implications for the evolution of cardiac arrhythmias such as atrioventricular heart block and modulated parasystole.

Effects of benzyltetrahydropalmatine on delayed after depolarization and triggered activity and on His-bundle electrogram and ECG in animal experiment.

Standard microelectrode techniques were used to study the effects of benzyltetrahydropalmatine (BTHP) on ouabain-induced delayed after depolarization (DAD) and triggered activity in isolated guinea papillary muscles. The results indicate that ouabain-induced DAD and triggered activity were abolished by BTHP 100 mumol/L. In anesthetized rabbit ECG heart rate was reduced in a dose-dependent manner from control value of 288 +/- 14 to 261 +/- 14 (BTHP 5 mg/kg) and 226 +/- 36 bpm (BTHP 10 mg/kg). P-R interval was prolonged. In His-bundle electrogram, H-V interval and V duration were not affected, but A-H interval was prolonged from 41 +/- 3 to 45 +/- 5 ms.

Theoretical computation of phase locking in embryonic atrial heart cell aggregates.

The effects of periodic stimulation of spontaneously beating aggregates of chick atrial heart cells are considered. Provided the effects of a single stimulus do not change the properties of the oscillation, and that the oscillation is re-established rapidly following a stimulus, this system can be modeled by one-dimensional finite difference equations. These equations employ experimentally generated phase resetting data that describe the effects of a single isolated stimulus at different phases of the oscillation. A complete analysis of the predicted dynamics is given over a broad range of stimulation frequencies and amplitudes. Prominent features of the dynamics include phase locking, bistability, chaos, and disappearance of Arnold tongues at large stimulation amplitudes. The fine details of the bifurcations are sensitive to properties of the phase resetting curves, and consequently, the observed bifurcations are not expected to be "universal" for larger stimulation amplitudes. Experimental traces show many correspondences with theoretical computations.

[Effects of benzyltetrahydropalmatine on action potentials of myocardium and transmembrane K+ and Ca2+ currents in Purkinje fibers].

Standard microelectrode and two-microelectrode voltage clamp techniques were used to study the effects of benzyltetrahydropalmatine (BTHP) on action potentials of isolated myocardium and transmembrane K+ and Ca2+ currents in Purkinje fibers. The effect of BTHP 3-100 mumol/L consisted of prolongation of the action potential duration and reduction of delayed rectifier current (Ik) in concentration-dependent manner. At concentration above 200 mumol/L, the contractile force of the isolated myocardium was depressed and in voltage clamp experiments the slow inward current (Isi) was reduced. These results suggest that the inhibition of Ik induced by BTHP was in relation to its anti-arrhythmic action.

[Effects of benzyltetrahydropalmatine on delayed after depolarization and triggered activity and rabbit His-bundle electrogram].

Standard microelectrode techniques were used to study the effects of benzyltetrahydropalmatine (BTHP) on ouabain-induced delayed after depolarization (DAD) and triggered activity in isolated guinea pig papillary muscles. The results indicate that ouabain-induced DAD and triggered activity were abolished by BTHP 100 mumols/L. In anesthetized rabbit ECG heart rates were reduced in a dose-dependent manner from its control value of 288 +/- 14 to 261 +/- 14 (5 mg/kg) and 226 +/- 36 bpm (BTHP 10 mg/kg). P-R interval was prolonged. In His-bundle electrogram, H-V interval and V duration were not affected, but A-H interval was prolonged from 41 +/- 3 to 45 +/- 5 ms.