Development of a diagnostic model for pre-washout screening of primary aldosteronism.

PURPOSE: Primary aldosteronism (PA) diagnosis is affected by antihypertensive drugs that are commonly taken by patients with suspected PA. In this study, we developed and validated a diagnostic model for screening PA without drug washout. METHODS: We retrospectively analyzed 1095 patients diagnosed with PA or essential hypertension. Patients were randomly grouped into training and validation sets at a 7:3 ratio. Baseline characteristics, plasma aldosterone concentration (PAC), and direct renin concentration (DRC) before and after drug washout were separately recorded, and the aldosterone-to-renin ratio (ARR) was calculated. RESULTS: PAC and ARR were higher and direct renin concentration was lower in patients with PA than in patients with essential hypertension. Furthermore, the differences in blood potassium and sodium concentrations and hypertension grades between the two groups were significant. Using the abbreviations potassium (P), ARR (A), PAC (P), sodium (S), and hypertension grade 3 (3), the model was named PAPS3. The PAPS3 model had a maximum score of 10, with the cutoff value assigned as 5.5; it showed high sensitivity and specificity for screening PA in patients who exhibit difficulty in tolerating drug washout. CONCLUSION: PA screening remains crucial, and standard guidelines should be followed for patients to tolerate washout. The PAPS3 model offers an alternative to minimize risks and enhance diagnostic efficiency in PA for those facing washout challenges. Despite its high accuracy, further validation of this model is warranted through large-scale clinical studies.

Bioinformatic analysis identified common pathogenetic processes between epilepsy and COVID-19.

OBJECTIVE: With the ongoing progression of SARS-CoV-2-induced COVID-19, the post-acute COVID-19 syndrome (long COVID) has garnered increasing attention as a novel multisystem disorder. Long COVID-19 has been shown to impact the nervous system, leading to various neurological manifestations, including epilepsy and seizures. Current studies have reported a significant increase in the prevalence and mortality rate of epilepsy in COVID-19 patients. Additionally, COVID-19 exacerbates seizures in patients with epilepsy. However, the mechanisms underlying the impact of COVID-19 on epilepsy remain elusive. This research focused on further identifying and elucidating the molecular mechanisms and biological processes underlying the induction of epilepsy by COVID-19 through bioinformatic methods. MATERIALS AND METHODS: We retrieved four gene expression datasets related to COVID-19 and epilepsy patients from the GEO and ArrayExpress databases. By crossing the major modules of weighted gene co-expression network analysis (WGCNA), the commonly expressed genes of epilepsy and COVID-19 were identified. By establishing the protein-protein interaction (PPI) network of the common genes, 20 hub genes were recognized through CytoHubba. Furthermore, functional enrichment and immune cell infiltration analyses were conducted to explore the potential mechanisms of COVID-19-related epilepsy. RESULTS: We identified a total of 373 common genes between the two diseases. The functional enrichment analysis revealed that the common genes were mainly involved in biological processes related to the immune response. Further analysis of the Hub genes revealed the important role of abnormal lipid metabolism in the crosstalk between COVID-19 and epilepsy. LASSO regression identified CD38 and PRKCA as the potential shared diagnostic candidates, which also exhibited excellent diagnostic value in the validation dataset. The immune infiltration analysis showed that activated dendritic cells (DCs) were positively correlated with the phenotypes of both diseases. CONCLUSIONS: This research revealed the potential mechanisms of COVID-19-related epilepsy, providing novel insights for the prevention, diagnosis, and clinical management strategies of COVID-19-related epilepsy.

H3K27M mutant diffuse midline glioma: a case report.

OBJECTIVE: Diffuse midline glioma with H3K27M mutation is a new tumor type of WHO central nervous system tumor classification. It often occurs in the midline structure and usually has a poor prognosis. CASE REPORT: A 38-year-old male patient presented with 2 years history of right limb with facial numbness, tumors in the left thalamic region and lateral ventricle was detected by imaging. The patient underwent the first surgery. RESULTS: The pathological examination results: Glioblastoma. He recovered well after surgery and received a total of 30 times of radiotherapy and temozolomide for one year. Fourteen months later, tumours were observed in the left thalamic region and left parieto-occipital lobe, the patient underwent the second operation. Immunohistochemistry showed: H3K27M(+). He experienced limitation of right limb movement after surgery and started taking oral apatinib 250 mg qd. After one-year, multiple tumors were found in the left brainstem, bilateral ventricles, bilateral basal ganglia, etc. The patient was given radiotherapy 7 times and then took apatinib 250 mg qd. Now the patient is still alive. CONCLUSIONS: H3K27M mutant diffuse midline glioma is characterized by diffuse infiltrative growth. Its pathological classification is diverse, imaging features lack specificity, and prognostic factors are complex. Traditional radiochemotherapy has limited effects, molecular targeted therapy, especially intervention of epigenetic regulation is being explored.

Dietary Fat Intake and Cognitive Function among Older Populations: A Systematic Review and Meta-Analysis.

OBJECTIVE: The associations between dietary fat intake and cognitive function are inconsistent and inconclusive. This study aimed to provide a quantitative synthesis of prospective cohort studies on the relationship between dietary fat intake and cognitive function among older adults. METHODS: PubMed, EMBASE, PsycINFO and Web of Science databases were searched for prospective cohort studies published in English before March 2018 reporting cognitive outcomes in relation to dietary fat intake. Four binary incident outcomes included were mild cognitive impairment (MCI), dementia, Alzheimer disease (AD) and cognitive impairment. The categories of dietary fat intake were based on fat consumption or the percentage of energy from fat consumption, including dichotomies, tertiles, quartiles and quintiles. The relative risk (RR) with the corresponding 95% confidence intervals (CIs) was pooled using a random effects model. RESULTS: Nine studies covering a total of 23,402 participants were included. Compared with the lowest category of consumption, the highest category of saturated fat intake was associated with an increased risk of cognitive impairment (RR = 1.40; 95% CI: 1.02-1.91) and AD (RR: 1.87, 95% CI: 1.09-3.20). The total and unsaturated fat intake was not statistically associated with cognitive outcomes with significant between-study heterogeneity. CONCLUSION: This study reported a detrimental association between saturated fat intake and cognitive impairment and mixed results between unsaturated fat intake and selected cognitive outcomes. Given the substantial heterogeneity in the sample size and methodology used across studies, the evidence presented here should be interpreted with caution.

Drug efficacies on bone mineral density and fracture rate for the treatment of postmenopausal osteoporosis: a network meta-analysis.

OBJECTIVE: Globally, a great number of elderly suffer from osteoporosis, especially postmenopausal women. Osteoporosis results in low bone mineral density (BMD) and high risk of fragility fracture. However, there is no defined strategy to select the most suitable anti-osteoporotic drugs for osteoporosis patients. Therefore, this study aims to select the most effective anti-osteoporotic drug for postmenopausal women with osteoporosis. MATERIALS AND METHODS: Literature search was conducted in PubMed, EMBASE, and the Cochrane Library. Raw data from the related randomized clinical trials were extracted. A pairwise and network meta-analysis model was utilized to assess the efficacy of ten drugs on the percentage change of BMD in the lumbar spine and total hip from baseline to one year of treatment. Risks of vertebral fracture and non-vertebral fracture were evaluated as well. We reported the effect size with a weighted mean difference (WMD) for continuous outcomes and odds ratio (OR) for dichotomous outcomes. All the drugs were ranked based on the surface under the cumulative ranking curve (SUCRA) value. Furthermore, the heterogeneity, consistency and publication bias of enrolled literature were assessed. RESULTS: With regard to lumbar spine BMD, the ten selected drugs all showed significant efficacy compared with placebo. In regard to total hip BMD and vertebral fracture, with the exception of calcitonin, the remaining nine drugs all showed significant efficacy compared with placebo. Six drugs - abaloparatide, alendronate, risedronate, strontium ranelate, teriparatide, and zoledronate - were significantly more effective compared with placebo for the treatment of non-vertebral fractures. As the SUCRA values indicated, abaloparatide performed the best on improving lumbar spine BMD, vertebral fracture and non-vertebral fracture, while denosumab was the best choice to improve total hip BMD. CONCLUSIONS: To sum up, abaloparatide, denosumab, and teriparatide showed the best efficacy for the treatment of postmenopausal osteoporosis, especially abaloparatide.

MiR-181b regulates atherosclerotic inflammation and vascular endothelial function through Notch1 signaling pathway.

OBJECTIVE: To explore the influences of micro ribonucleic acid (miR)-181b on the inflammation and vascular endothelial function in atherosclerosis (AS), and its specific molecular regulatory mechanism. MATERIALS AND METHODS: 44 apolipoprotein E (ApoE)-/- 7 weeks old male rats were randomly divided into normal diet group (NC group) and AS model group (high-fat diet feeding). Rat aorta was dissected and the serum sample was collected in both groups. The serum levels of inflammatory factors in both groups were detected via enzyme-linked immunosorbent assay (ELISA). The mRNA levels of miR-18b and Notch1 were detected via Reverse Transcription-Polymerase Chain Reaction (RT-PCR). Moreover, aortic endothelial cells were extracted from AS rats. The mir-18b binding target gene was analyzed via bioinformatics and further verified by the Luciferase reporter gene assay. The protein expressions of miR-18b and Notch1 in endothelial cells transfected with miR-181b mimic or inhibitor were detected. Influence of miR-181b on vascular endothelial indexes was also detected. RESULTS: Compared with those in the NC group, the serum levels of interleukin-6 (IL-6), IL-10, IL-13 and tumor necrosis factor-α (TNF-α) in the AS group significantly increased (p<0.05). The mRNA level of miR-18b in AS plaques was significantly lower than that in NC arterial tissues. Conversely, Notch1 level in AS plaques was markedly higher than that in the NC arterial tissues (p<0.05), with the mean difference of 2.12 and 2.82 folds (p<0.05). According to the Pearson correlation analysis, there was a significant negative correlation between mRNA levels of miR-181b and Notch1 in AS tissues (r=-0.65, p=0.014). The bioinformatics analysis showed that there were complementary binding sites between miR-181b and Notch1. The Luciferase reporter gene assay confirmed the presence of direct binding sites between miR-181b and Notch1. Western blotting revealed that the overexpression of miR-181b downregulated Notch1 and hs-CRP, but upregulated BNP (p<0.05). Opposite trends were obtained by miR-181b knockdown (p<0.05). CONCLUSIONS: The decline in the miR-181b expression may be an important factor in AS plaque formation and vascular endothelial injury by regulating Notch1.

Risk of systemic lupus erythematosus in patients with human papillomavirus infection: a population-based retrospective cohort study.

BACKGROUND: Viral infection contributing to systemic lupus erythematosus (SLE) development has been largely reported. However, the SLE risk in patients with human papillomavirus (HPV) infection is unknown. METHODS: Data were retrieved from the Longitudinal Health Insurance Database (2000) in Taiwan. We identified 43,567 patients with HPV infection and 174,268 age- and sex-matched uninfected controls from 2002 to 2012. Individuals were followed up from index date (first date of diagnosis with HPV) until the occurrence of SLE, at the end of the study (December 2013), or when they were withdrawn from the insurance program. The incidence rate ratio (IRR) was calculated using the univariate Poisson regression. The adjusted hazard ratios (aHRs) were calculated, and sensitive and subgroups analyses were also conducted. RESULTS: Compared with the non-HPV controls, the IRR of SLE in HPV patients was 1.52 (95% confidence interval (CI): 1.09-2.12). The risk of SLE in HPV-infected individuals was significantly high (aHR: 1.48, 95% CI: 1.06-2.06) after adjusting for age, sex, and comorbidities. Men aged between 16 and 45 years were more susceptible to developing SLE (aHR: 21.57, 95% CI: 2.52-184.60, p = 0.0051). CONCLUSION: Our study showed a significantly higher risk of SLE among HPV-infected patients, especially in men aged between 16 and 45 years.

High expression of GAS5 promotes neuronal death after cerebral infarction by regulating miR-365a-3p.

OBJECTIVE: To investigate whether growth arrest specific5 (GAS5) could regulate the expression of DCDC2 through competitive binding to miR-365a-3p, thus leading to increased neuronal death in cerebral cortex. MATERIALS AND METHODS: The expression levels of GAS5 and DCDC2 in cerebral cortical neurons of mice with mouse middle cerebral artery occlusion (MCAO) cerebral infarction and in control mice were detected by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). Meanwhile, the expression levels of GAS5 and DCDC2 in primary neurons cultured in vitro were also detected by qRT-PCR. The effects of GAS5 and DCDC2 on neuronal death were evaluated by calculating the cerebral infarct area by Tunel assay and TTC staining. Dual luciferase reporter assays were performed to detect the binding of miR-365a-3p to GAS5 and DCDC2. Western blot was applied to detect the protein expression of DCDC2, Bcl-2 and Bax after overexpression and knockdown of GAS5. RESULTS: The expression of GAS5 and DCDC2 were significantly higher in cerebral cortical neurons and primary cultured neurons in vitro than those in control mice, respectively. Inhibition of GAS5 and DCDC2 in primary neurons decreased the neuronal cells death rate, while overexpression of GAS5 and DCDC2 increased the cell death rate. The dual luciferase reporter gene results showed that GAS5 regulated the expression of DCDC2 through competitive binding of miR-365a-3p thus forming a GAS5/miR-365a-3p/DCDC2 regulatory network. In addition, GAS5 inhibited Bcl-2 while promoting the expression of Bax. CONCLUSIONS: High expression of GAS5 could promote neuronal death after cerebral infarction in mice, possibly through competitive binding to miR-365a-3p and promoting the expression of DCDC2.

STEEL participates in fracture healing through upregulating angiogenesis-related genes by recruiting PARP 1.

OBJECTIVE: To explore the effect of STEEL on fracture healing and its underlying mechanism. PATIENTS AND METHODS: A total of 31 patients with long bone fracture and who received reoperation because of bone nonunion, delayed union or healing disorder in the Wuxi Nine Hospital Affiliated to Soochow University from July 2016 to February 2018 were selected. The bone callus at the fracture site was collected from each patient during the reoperation. QRT-PCR (Quantitative Real-Time Polymerase Chain Reaction) was used to detect STEEL expression in the callus tissues of the treatment group (bone nonunion or delayed union) and the control group. In addition, we measured the number of blood vessels in the fracture tissues by immunohistochemistry. After the construction of tibial fracture model in mice, STEEL expression and the total number of blood vessels in the treatment group (sawing treatment) and the control group (sham operation) were detected, respectively. For in vitro experiments, CCK-8 (cell counting kit-8) assay was performed to detect cell proliferation after knockdown or overexpression of STEEL in the vascular endothelial cells. The binding condition of STEEL and its interacting proteins were detected by RIP (RNA binding protein immunoprecipitation), and the binding of PARP 1 [poly (ADP-ribose) polymerase 1] with gene promoter was observed by ChIP (chromatin immunoprecipitation assay). Western blot was used to detect the expression level of VEGF (vascular endothelial growth factor). RESULTS: STEEL expression and the vascular density in the callus tissues of the treatment group were significantly lower than those of the control group. Downregulated STEEL remarkably decreased the proliferation ability of HUVEC cells. Meanwhile, the vascular density was also significantly decreased in mice with a tibial fracture. Overexpressed STEEL obtained the opposite results. STEEL could interact with PARP 1 to regulate expressions of downstream genes. Moreover, STEEL could also promote angiogenesis by elevating VEGF expression. CONCLUSIONS: We showed that STEEL expression could partly represent the angiogenesis of fracture sites. Moreover, it promoted angiogenesis by elevating VEGF expression.

Correlations of MC4R and MSH2 expression with obesity in colon cancer patients.

OBJECTIVE: To investigate the correlations of MC4R and MSH2 with adult obesity, a total of 46 patients with early-stage colon cancer were treated in our hospital between February 2008 and February 2009 and were enrolled. PATIENTS AND METHODS: Venous blood was regularly drawn from subjects of the observation group and 48 healthy subjects for 6 years. Expression levels of MC4R and MSH2 genes were tested using quantitative polymerase chain reaction analyses, and the ensuing proteins were determined using enzyme-linked immunosorbent assays and Western blotting and immunohistochemical analyses. Finally, correlations with body mass index (BMI) and the presence of colon cancer were identified using multivariate analyses. RESULTS: Compared with the control group, MSH2 mRNA and protein expression increased significantly over time (p < 0.05) in patients with colon cancer. Moreover, MSH2 expression was correlated with colon cancer progression, and MC4R mRNA and protein expression increase concurrently in comparison with the control group (p < 0.05). Also, the mean BMI among patients with colon cancer was 30.8, whereas that among control subjects was only 21.4. These data indicate a relationship between BMI and colon cancer. CONCLUSIONS: Expression of MSH2 in patients with colon cancer may promote the expression of the obesity gene MC4R, potentially contributing to body weight gains.

Sumoylation in Lens Differentiation and Pathogenesis.

Sumoylation, a post-translational modification discovered over a decade ago, turns out to be a very important regulatory mechanism mediating multiple cellular processes. Recent studies from our laboratory and others also revealed that it plays a crucial role in regulating both differentiation and pathogenesis of the ocular lens. This review will summarize these progresses.

Molecular Cloning and Developmental Expression Patterns of the Striatin Gene Encoding A Member of the Regulatory Subunits for the Protein Serine/Threonine Phosphatase-2A in Fish.

PURPOSE: The protein phosphatase-2A (PP-2A) is one of the most important serine/threonine phosphatases in eukaryotes. The holoenzyme of PP-2A consists of three subunits: a scaffold A subunit, a catalytic C subunit and a regulatory B subunit. While both A and C subunits are coded by two different genes, the B subunits exist in 26 or more isoforms which are encoded by at least 15 different genes. Previous studies have shown that besides regulating specific PP-2A activity, various B subunits may have other functions. To explore the possible roles of the regulatory subunits of PP-2A in vertebrate development, we have cloned the gene encoding goldfish striatin, a member of the B'" family regulatory subunits for PP-2A, and determined their tissue-specific and temporal expression patterns. METHODS: The cDNA cloning was conducted with RT-PCR-based RACE. The mRNA expression levels for the goldfish striatin were analyzed with RT-PCR. The expression levels of the striatin protein from goldfish were determined with Western blot analysis. The semi-quantitation of the mRNA and protein expression levels was conducted with the software of U-scanning. RESULTS: Our study revealed that the full length cDNA for striatin consists of 2965 bp coding for a deduced protein of 769 amino acids, which bears a very high level of amino acid sequence identity with the homolog protein from other species. The striatin mRNA is highly expressed in the kidney, to a less degree in brain, fin, muscle, liver, ovary and gill, and the lowest in testis and heart. Similar pattern of protein expression is detected in the above 9 tissues. During the development of goldfish, the striatin mRNA maintains a relatively high level at the 2-cell, multiple cell and blastula stages. Then, it drops down substantially at gastrula stage and fluctuates around this level in the next 8 different stages. At the protein level, the striatin maintained higher level from 2-cell to gastrula stages, then decreased at neurula and optic vesicle stages, and gradually increased again to peak at eye pigmentation stage, then slightly decreased in the next few stages of development. CONCLUSIONS: Our results suggest that the striatin may play an important role in regulating goldfish development and adult tissue homeostasis. While the former function may or may not occur through PP- 2A functions, the later function appears to occur via PP-2A activity.

Regulation of ATRIP protein abundance by RAD9 in the DNA damage repair pathway.

Genotoxic stress activates checkpoint signaling pathways that activate the checkpoint kinases ATM and ATR, halt cell cycle progression, and promote DNA repair. A number of proteins act in concert with ATR to phosphorylate Chk1, including RAD17, the RAD9-RAD1-HUS1 complex, ATR/ATRIP and TopBp1. However, how these proteins involved act in concert with one another to propagate and maintain the checkpoint response is not well understood. Here, we reported that upregulation of RAD9 protein increased the quantity of ATRIP, suggesting that RAD9 activation will induce more efficient accumulation of ATRIP in vivo. Furthermore, the DNA damage-induced ATRIP foci formation was faster in the mRad9-/- ES cells. Also, ATRIP interacts specifically with RAD9, but not HUS1 and RAD1. Taken together, we suggested that RAD9 could affect both the ATRIP protein levels and DNA damage-induced ATRIP foci formation. Thus, we propose a role of RAD9 in the ATR-Chk1 pathway that is necessary for successful formation of the damage-sensing complex and DNA damage checkpoint signaling.

Regulation of Eye Development by Protein Serine/Threonine Phosphatases-1 and -2A.

The protein serine/threonine phosphatases-1 and -2A are major cellular phosphatases, playing a fundamental role in organisms from prokaryotes to eukaryotes. They contribute to 90% dephosphorylation in eukaryote proteins. In the eye, both phosphatases are highly expressed and display important functions in regulating normal eye development. Moreover, they are implicated in pathogenesis through modulation of stress-induced apoptosis. Here we review the recent progresses on these aspects.

The Role of αA-Crystallin in Experimental Autoimmune Uveitis.

Uveitis refers to a group of ocular inflammatory diseases that can lead to blindness. For years, researchers have been trying to decipher the underlying mechanisms and develop therapeutic strategies using the model of experimental autoimmune uveitis (EAU). Recently, αA-crystallin has been found to be upregulated in EAU and can even ameliorate its severity through different mechanisms, suggesting its use as a potent therapeutic factor against uveitis. Here we review the protective role of αA-crystallin and discuss its functional mechanisms in EAU.

Clinical and experimental observation of the length of time of respiratory phase: a preliminary study.

OBJECTIVE: Our goal was to observe the duration of inhaling and exhaling as well as the inhalation to exhalation conversion in patients with lung cancer and experimental dogs. MATERIALS AND METHODS: We recorded lung and tumor respiratory motion with X-ray camera, in five patients with lung cancer as well as in five experimental dogs. We made random observation of breathing cycle inhalation duration, exhalation duration, and inhalation to exhalation conversion within each lung cancer patients and within each of the five animals. RESULTS: Respiratory inhalation duration of each dog and human > exhalation duration > exhale to inhale conversion length > inhalation to exhalation conversion length. During the four breathing cycles, the total respiratory duration differs, and the length of the same breathing phase is inconsistent. CONCLUSIONS: The measurement of early stage breathing duration cannot be representative of breathing duration of the late stage. Radiation treatment planning system based on the pre-computed tomography scanning on the basis of early stage, there will be some radiation dose errors.

Prognostic value of clinical and immunological markers in acute phase of SFTS virus infection.

SFTS virus (SFTSV) is a novel bunyavirus that causes severe fever with thrombocytopenia syndrome (SFTS), an emerging infectious disease that occurred in China in recent years, with an average case fatality rate of 10-12%. Intervention in the early clinical stage is the most effective measure to reduce the mortality rate of disease. To elucidate the natural course of and immune mechanisms associated with the pathogenesis of SFTSV, 59 laboratory-confirmed SFTS patients in the acute phase, who were hospitalized between October 2010 and September 2011, were enrolled in this study, and the patients sera were dynamically collected and tested for SFTSV viral RNA load, 34 cytokines or chemokines and other related laboratory parameters. All clinical diagnostic factors in the acute phase of SFTS were evaluated and assessed. The study showed that the severity of the disease in 11 (18.6%) patients was associated with abdominal pain (p 0.007; OR = 21.95; 95% CI, 2.32-208.11) and gingival bleeding (p 0.001; OR=122.11; 95% CI, 6.41-2328). The IP-10, TNF-α, IL-6, IL-10, granzyme B and HSP70 levels were higher over the 7-8 days in severe cases, accompanied by altered AST, CK and LDH levels. HSP70 (p 0.012; OR=8.29; 95% CI, 1.58-43.40) was independently correlated with the severity of the early acute phase of SFTSV infection. The severity of SFTS can be predicted based on the presence of symptoms such as abdominal pain and gingival bleeding and on the level of HSP70 in the acute phase of the disease.

ERK signaling pathway regulates embryonic survival and eye development in goldfish, Carassius auratus.

The extracellular signal-regulated kinase (ERK) is one of the three major types of mitogen-activated protein kinases. Previous studies showed that ERKs mediate various signaling pathways for cell proliferation, differentiation, survival and transformation in mammals. In the present study, we use goldfish as a model system and demonstrate that ERK kinases play important roles in promoting embryonic survival and regulate development of eye and trunk in vertebrates. ERKs are highly expressed in multiple tissues including lens epithelial cells, lens fiber cells, retina, brain, muscle and heart of adult goldfish. Injection of the dominant negative ERK mutant (DNM-ERK) into the fertilized eggs of goldfish significantly inhibited ERK activity at blastula stage, and completely blocked ERK activity at gastrula and later stages. As a result, the blastula cells were induced into apoptosis, and majority of the injected embryos were lethal at embryonic stages. At the molecular level, inhibition of ERK activity by DNM-ERKs suppressed phosphorylation of Bad at Ser-112 to promote apoptosis. Similar results were observed when MEK activity was inhibited by U0126 treatment. The survived embryos display significant abnormality in the phenotypes of both eye and trunk. Associated with the abnormality in the eye development, phosphorylation in Pax-6 and expression of HSF4 were significantly decreased and expression of the ß-crystallin gene was also downregulated. These results provide novel information regarding the roles of ERKs in regulating vertebrate development.

p53 directly regulates αA- and ßA3/A1-crystallin genes to modulate lens differentiation.

It is well established that the tumor suppressor p53 plays major roles in regulating apoptosis and cell cycle progression. In addition, recent studies have demonstrated that p53 is actively involved in regulating cell differentiation in muscle, the circulatory system and various carcinoma tissues. We have recently shown that p53 also controls lens differentiation. Regarding the mechanism, we reveal that p53 directly regulates c-Maf and Prox1, two important transcription factors to control cell differentiation in the ocular lens. In the present study, we present further evidence to show that p53 can regulate lens differentiation by controlling expression of the differentiation genes coding for the lens crystallins. First, the αA and ßA3/A1 gene promoters or introns all contain putative p53 binding sites. Second, gel mobility shifting assays revealed that the p53 protein in nuclear extracts from lens epithelial cells directly binds to the p53 binding sites found in these crystallin gene promoters or introns. Third, exogenous wild type p53 induces dose-dependent expression of the luciferase reporter gene driven by different crystallin gene promoters and the exogenous dominant negative mutant p53 causes dose-dependent inhibition of the same crystallin genes. Fourth, ChIP assays revealed that p53 binds to crystallin gene promoters in vivo. Finally, in the p53 knockout mouse lenses, expression levels of various crystallins were found down-regulated in comparison with those from the wild type mouse lenses. Together, our results reveal that p53 directly regulates expression of different sets of genes to control lens differentiation.

Rapid downregulation of programmed death-1 and interferon-γ-inducible protein-10 expression is associated with favourable outcome during antiviral treatment of chronic hepatitis B.

The dynamics of programmed death-1 (PD-1) as well as cytokine/chemokine expression and its correlation with virological response in patients with chronic hepatitis B (CHB) is unclear. This study was conducted in 29 treatment-naïve patients undergoing telbivudine treatment for 52 weeks. The results showed that PD-1 expression on both CD4+ and CD8+ T cells was positively correlated with hepatitis B virus (HBV) DNA levels (r = 0.621, P < 0.0001; r = 0.548, P = 0.002, respectively), and in virological responders, this decrease was directly correlated with a decrease in HBV DNA levels (r = 0.664, P = 0.002; r = 0.572, P = 0.01, respectively). Furthermore, at the end of 52 weeks, in virological responders, the decreased rate in the frequency of PD-1+ CD8+ T cells was significantly higher than in non-virological responders (58.3% vs 25.7%, P = 0.001), and at weeks 24 and 52, in virological responders, PD-1 expression on CD4+ and CD8+ T cells was lower than in non-virological responders (P = 0.01 and P = 0.035; P < 0.0001 and P < 0.0001, respectively). In 34 cytokines/chemokines detected in serum, IP-10 expression was positively correlated with viral load, level of ALT and PD-1 expression on CD8+ and CD4+ T cells at baseline (r = 0.36, P = 0.055, r = 0.635, P < 0.0001, r = 0.414, P = 0.026, and r = 0.402, P = 0.030, respectively). Moreover, the decrease in IP-10 in serum directly correlated with a decrease in ALT levels (r = 0.751, P < 0.0001). At weeks 24 and 25, IP-10 expression was significantly lower than baseline in virological responders (both P = 0.005); however, this was not observed in nonresponders. Based on the above findings, PD-1 and IP-10 may be used as predictors for virological response, and blockade of their pathway may improve the outcome of patients with CHB.