[Discussion on the composition and implementation of diagnosis and treatment strategies for whole field pain management strategy].

Pain is the fifth major vital sign, and chronic pain is a large category of diseases that affects health seriously. At present, the incidence of chronic pain is high, but the overall treatment satisfaction is low. It is necessary to continuously optimize pain diagnosis and treatment strategies and improve the connotation of pain management. Based on the clinical practice of our pain center, combined with relevant literature, the article proposes a diagnosis and treatment strategy of "whole field pain management" should be carried out from the four dimensions of feeling, emotion, cognition, and behavior. Innovative digital pain diagnosis and treatment technologies such as VR/MR and brain-computer interface are used to regulate emotional, cognitive, and behavioral regulation, and combined with lifestyle changes, rehabilitation physiotherapy, drugs, and minimally invasive interventional therapy to constitute a " whole field pain management strategy" to explore the new development direction of further improving the management of chronic pain.

[Impact of different diagnostic criteria for assessing mild micro-hepatic encephalopathy in liver cirrhosis: an analysis based on a prospective, multicenter, real-world study].

Objective: To compare the differences in the prevalence of mild micro-hepatic encephalopathy (MHE) among patients with cirrhosis by using the psychometric hepatic encephalopathy score (PHES) and the Stroop smartphone application (Encephal App) test. Methods: This prospective, multi-center, real-world study was initiated by the National Clinical Medical Research Center for Infectious Diseases and the Portal Hypertension Alliance and registered with International ClinicalTrials.gov (NCT05140837). 354 cases of cirrhosis were enrolled in 19 hospitals across the country. PHES (including digital connection tests A and B, digital symbol tests, trajectory drawing tests, and serial management tests) and the Stroop test were conducted in all of them. PHES was differentiated using standard diagnostic criteria established by the two studies in China and South Korea. The Stroop test was evaluated based on the criteria of the research and development team. The impact of different diagnostic standards or methods on the incidence of MHE in patients with cirrhosis was analyzed. Data between groups were differentiated using the t-test, Mann-Whitney U test, and χ (2) test. A kappa test was used to compare the consistency between groups. Results: After PHES, the prevalence of MHE among 354 cases of cirrhosis was 78.53% and 15.25%, respectively, based on Chinese research standards and Korean research normal value standards. However, the prevalence of MHE was 56.78% based on the Stroop test, and the differences in pairwise comparisons among the three groups were statistically significant (kappa = -0.064, P < 0.001). Stratified analysis revealed that the MHE prevalence in three groups of patients with Child-Pugh classes A, B, and C was 74.14%, 83.33%, and 88.24%, respectively, according to the normal value standards of Chinese researchers, while the MHE prevalence rates in three groups of patients with Child-Pugh classes A, B, and C were 8.29%, 23.53%, and 38.24%, respectively, according to the normal value standards of Korean researchers. Furthermore, the prevalence rates of MHE in the three groups of patients with Child-Pugh grades A, B, and C were 52.68%, 58.82%, and 73.53%, respectively, according to the Stroop test standard. However, among the results of each diagnostic standard, the prevalence of MHE showed an increasing trend with an increasing Child-Pugh grade. Further comparison demonstrated that the scores obtained by the number connection test A and the number symbol test were consistent according to the normal value standards of the two studies in China and South Korea (Z = -0.982, -1.702; P = 0.326, 0.089), while the other three sub-tests had significant differences (P < 0.001). Conclusion: The prevalence rate of MHE in the cirrhotic population is high, but the prevalence of MHE obtained by using different diagnostic criteria or methods varies greatly. Therefore, in line with the current changes in demographics and disease spectrum, it is necessary to enroll a larger sample size of a healthy population as a control. Moreover, the establishment of more reliable diagnostic scoring criteria will serve as a basis for obtaining accurate MHE incidence and formulating diagnosis and treatment strategies in cirrhotic populations.

[Epidemiological distribution of genotypes and sub-genotypes of hepatitis B virus in 15 ethnic groups in China].

Objective: To understand the distribution of genotypes and sub-genotypes of HBV in different ethnic groups in China. Methods: The HBsAg positive samples were selected by stratified multi-stage cluster sampling from the sample base of national HBV sero-epidemiological survey in 2020 for the amplification of S gene of HBV by nested PCR. A phylogeny tree was constructed to determine the genotypes and sub-genotypes of HBV. The distribution of genotypes and sub-genotypes of HBV were analyzed comprehensively by using laboratory data and demographic data. Results: A total of 1 539 positive samples from 15 ethnic groups were successfully amplified and analyzed, and 5 genotypes (B, C, D, I and C/D) were detected. The proportion of genotype B was higher in ethnic group of Han (74.52%, 623/836), Zhuang (49.28%, 34/69), Yi (53.19%, 25/47), Miao (94.12%, 32/34), Buyi (81.48%, 22/27). The proportions of genotype C were higher in ethnic groups of Yao (70.91%, 39/55). Genotype D was the predominant genotype in Uygur (83.78%, 31/37). Genotype C/D were detected in Tibetan (92.35%,326/353). In this study, 11 cases of genotype I were detected, 8 of which were distributed in Zhuang nationality. Except for Tibetan, sub-genotype B2 accounted for more than 80.00% in genotype B in all ethnic groups. The proportions of sub-genotype C2 were higher in 8 ethnic groups, i.e. Han, Tibetan, Yi, Uygur, Mongolian, Manchu, Hui and Miao. The proportions of sub-genotype C5 were higher in ethnic groups of Zhuang (55.56%, 15/27) and Yao (84.62%, 33/39). For genotype D, sub-genotype D3 was detected in Yi ethnic group and sub-genotype D1 was detected in both Uygur and Kazak. The proportions of sub-genotype C/D1 and C/D2 in Tibetan were 43.06% (152/353) and 49.29% (174/353). For all the 11 cases of genotype I infection, only sub-genotype I1 was detected. Conclusions: Five genotypes and 15 sub-genotypes of HBV were found in 15 ethnic groups. There were significant differences in the distribution of genotypes and sub-genotypes of HBV among different ethnic groups.

A prognostic model based on ferroptosis-related long non-coding RNA signatures and immunotherapy responses for non-small cell lung cancer.

OBJECTIVE: Non-small cell lung cancer (NSCLC) ranks high in the incidence of malignant tumors, with limited treatment options and poor prognosis. Ferroptosis is a newly discovered cell death mechanism based on iron and reactive oxygen species (ROS). The role of ferroptosis-related long non-coding RNAs (lncRNAs) and associated prognostic mechanisms in NSCLC require investigation. MATERIALS AND METHODS: We constructed a prognostic multi-lncRNA signature based on ferroptosis-related differentially expressed lncRNAs in NSCLC. The levels of ferroptosis-related lncRNA in normal lung cells and lung adenocarcinoma cells were verified by RT-PCR. RESULTS: We identified eight differentially expressed lncRNAs associated with NSCLC prognosis. The expression of AC125807.2, AL365181.3, AL606489.1, LINC02320, and AC099850.3 was upregulated, while SALRNA1, AC026355.1, and AP002360.1 were downregulated in NSCLC cell lines. Kaplan-Meier analysis showed that a high-risk patient group was associated with poor NSCLC prognosis. A risk assessment model based on ferroptosis-related lncRNAs was superior to NSCLC prognosis based on traditional clinicopathological features. Gene Set Enrichment Analysis (GSEA) identified immune- and tumor-related pathways in low-risk group patients. In addition, The Cancer Genome Atlas (TCGA) showed that T cell function during APC co-inhibition, APC co-stimulation, chemokine receptor (CCR), MHC class I, parainflammation, T cell co-inhibition, and check-point expression differed significantly between low- and high-risk groups. M6A-related mRNA comparisons between these groups also revealed significant differences in ZC3H13, RBM15, and METTL3 expression. CONCLUSIONS: Our new model of lncRNA-associated ferroptosis effectively predicted NSCLC prognoses.

Construction of a hypoxic immune microenvironment associated gene-based model for prognosis prediction of lung adenocarcinoma.

OBJECTIVE: Lung adenocarcinoma (LUAD) accounts for the majority of cancer deaths worldwide, with a high incidence rate and mortality. It is highly important to develop biomarker model to accurately predict the prognosis. MATERIALS AND METHODS: RNA-Seq data and clinical follow-up data of LUAD were downloaded from The Cancer Genome Atlas (TCGA) database. Hypoxia-related gene sets were collected from the Gene Set Enrichment Analysis (GSEA) website. A gene signature model was established using the Limma package in the R software, univariate and multivariate survival analyses, and least absolute shrinkage and selection operator (LASSO) algorithms. RESULTS: Two hypoxia subtypes (C1 and C2) were classified according to the expressions of 55 prognostic hypoxic-related genes. Differentially expressed genes (DEGs) between two hypoxia subtypes and immune group were analyzed. Then, 390 DEGs related to hypoxic immune microenvironment were filtered. According to hypoxia type and immune type, the samples were classified into hypoxia-high & immune-low group, hypoxia-low & immune-high group. Based on these differentially expressed genes (DEGs), a 5-genes signature model, which showed a stable prediction performance on datasets of different platforms and immunotherapy datasets, was finally developed. Meanwhile, it demonstrated a better performance compared with other existing models. The AUC of the 5-gene signature was high in both the training dataset and 4 independent validation datasets and was confirmed as a clinical feature-independent prognostic model. CONCLUSIONS: This study developed a hypoxic immune microenvironment associated gene-based model for prognostic prediction of LUAD, providing clinicians with a reliable prognostic assessment tool and facilitating clinical treatment decision-making.

[Effect of China Children's Asthma Action Plan on exercise in school-age children with bronchial asthma].

Objective: To investigate the effect of China Children's Asthma Action Plan (CCAAP) on the exercise status of school-age children with asthma. Methods: We included 400 school-age asthmatic children as research objects from CCAAP asthma management platform of the Affiliated Hospital of Qingdao University during March 1, 2018 to February 28, 2021 by simple random sampling method. The questionnaires of basic information and international physical activity were applied through WeChat or face to face investigation to collect the basic information and exercise status of the object. There were 346 valid questionnaires included in the study to compare the differences in exercise status and incidence of exercise-related asthma-like symptoms between the good and poor CCAAP application groups. Results: There were 232 (67.05%) and 114 (32.95%) cases in good and poor CCAAP application group, respectively. Age, female proportion and BMI of good CCAAP application group were (8±2) years, 47.0% (109/232) and (19.79±2.32) kg/m2, respectively, no statistic difference comparing to poor CCAAP application group [(8±2) years, 46.5% (53/114) and (19.87±2.43) kg/m2, respectively] (all P values>0.05). In good CCAAP application group, 30.18% (70/232) achieved the standard of moderate (high) intensity exercise per day, no statistic difference comparing to poor CCAAP application group [29.82% (34/112)] (P=0.947); 31.90% (74/232) participated in high-intensity exercise per week, higher than that of poor CCAAP application group [17.54% (20/112)] (P=0.005); incidence of exercise-related asthma-like symptoms was 19.83% (46/232), lower than that of poor CCAAP application group [29.82% (34/112)] (P=0.038). Conclusion: CCAAP promotes the exercise of school-age children with asthma.

Development of a prognostic model based on an immunogenomic landscape analysis of colorectal cancer.

Screening and therapeutic programs for colorectal cancer (CRC) are invasive or not effective and unable to meet patient needs. Major advances in immunogenomics may change this status but need more exploration. Differentially expressed genes and immune-related genes (IRGs) were identified by computational methods. A prognostic model was established and validated based on survival-related IRGs via stepwise multivariate Cox regression analysis. Nine IRGs were selected and identified as survival-related genes. A 7-gene prognostic model could offer a preliminary and valid determination of risk in CRC patients. The area under the curve of the receiver operating characteristic was 0.672. The 7-gene prognostic model might be used as a novel prognostic tool in CRC patients.

[A follow-up study of abnormal mutation in neonatal deafness gene screening].

Objective: To screen, diagnose and follow up the abnormal mutation in the gene screening of neonatal deafness. Methods: A total of 24161 newborns born in Zhuhai Maternal and Child Health Hospital from February 1, 2015 to January 31, 2008 were screened for hearing and deafness genes, and audiological screening, diagnosis and 1-3 years follow-up were carried out for the newborns with positive gene screening. Results: There were 991 cases of deafness gene mutation (533 males and 458 females), and the rate of abnormal mutation was 4.10%(991/24 161). Among them, 921 cases were single heterozygous mutation, 130 cases were failed in primary hearing screening, 11 cases were failed in secondary hearing screening, 8 cases were abnormal in audiological diagnosis finally. In these 8 cases, 3 were diagnosed as otitis media and passed audiological follow-up after cure, 2 cases of single ear sensorineural injury caused by high-risk factors, passed after close audiological follow-up, and the other 3 cases were closely audiological follow-up while none of them were successfully sequenced. All of them were moderate to severe sensorineural deafness, 1 case was heterozygous mutation at 3 loci of GJB2(c.235delC,c.408C>A,c.134G>A), 1 case was heterozygous mutation at 2 loci of GJB2(c.235delC, c.109G>A), and 1 case was single heterozygous mutation of GJB2(c.235delC). The remaining 913 cases who passed the primary screening, secondary screening or hearing diagnosis were followed up for 1 to 3 years. Three cases of multiple heterozygous mutation were found in gene screening(2 cases were SLC26A4 2168A>G, IVS7-2A>G, 1 case was GJB2 c.176_191del 16bp, c.299_300del AT), all of them passed both primary and secondary hearing screening. In these 3 cases, the final audiological diagnosis was moderate sensorineural deafness in both ears, with no improvement in the follow-up of 1-3 years. There were 9 monogenic homozygous mutations, 7 failed in primary hearing screening, 3 failed in secondary hearing screening and also failed in audiological diagnosis and 1-3 years' audiological follow-up, all of whom were GJB2 c.235 del C homozygous mutations, and one of whom had a definite family history of deafness. The remaining 6 cases of homozygous mutation diagnosed by primary screening, secondary screening or hearing diagnosis were GJB2 c109G>A homozygous mutation, and passed the 1-3 years' hearing follow-up. 58 children with mtDNA mutations, including 2 with 12S rRNA 1494C>T homozygous mutation, 47 with 1555A>G homozygous mutation, and 9 with 1555A>G heterozygous mutation, all passed the primary or secondary hearing screening, and were instructed to ban ototoxic drugs for the whole life, and passed the 1-3 years' hearing follow-up. Conclusions: The audiological follow-up of children with monogenic heterozygous mutations in deafness gene screening is generally normal. In case of abnormality, the influencing factors such as otitis media should be excluded at first. In case of unexplained moderate to severe sensorineural deafness, the whole-gene sequencing should be performed to find possible pathogenic factors. The children with homozygous mutation or compound heterozygous mutation in gene screening, most of whom show different degrees of hearing loss, should be followed up for a long time, and provide parents with scientific and reasonable genetic counseling according to the mutation genes and loci,. The hearing of drug-induced deafness gene carriers is normal after birth. Parents should be advised to strengthen prevention and follow-up is generally enough.

[Congenital ear deformity screening and non-invasive correction effect analysis].

Objective:To synchronously perform external auricle examination during neonatal hearing screening, follow up auricle deformity with neonatal disease screening system, and calculate the incidence of auricle deformity, self-healing rate, correction rate, incidence of complications and the relationship with hearing loss in Zhuhai area. Method:According to the diagnostic criteria of auricle deformity, the newborns in Zhuhai Maternal and Child Health Hospital were examined on the spot within 2 months. The deformity auricle was registered and uploaded into the newborn hearing screening system. The newborns were followed up by short message notification 7 days after birth, and then compared with the photo uploading system again. At 14 days, the ears of those who could not self-heal were went on non-invasive correction, and collect of relevant data for summary analysis. Result:Among the 1 073 newborns(2 146 ears), 26(37 ears) with malformed ears were treated with auricular pattern correction.The corrective rates of newborns less than 14 days, 14-30 days and 31-60 days were 95%, 90% and 87% respectively, and the incidence of complications were 50%, 58% and 69%, respectively. Conclusion:The incidence of auricular deformities in neonates is high. The earlier correction the better. The ear deformity can be detected at the earliest stage and missed diagnosis can be avoided by simultaneous hearing screening and ear deformity screening. During the window period of 7-14 d, by monitoring the self-healing rate of the affected ear excessive medical correction can be avoided. By hearing screening system statistics, ear shape malformation is not directly related to hearing loss.

[Hearing assessment and follow-up study of aeonatal deafness gene screening homozygous mutation infants].

Objective:To analyze the hearing assessment characteristics and follow-up of some deafness gene screening homozygous infants in Zhuhai. Method:The clinical data of 28 newborns with homozygous mutations transferred to Zhuhai Maternal and Child Health Hospital from Feb. 1, 2015 to Oct. 25, 2018 in hospitals of Zhuhai City were retrospectively analyzed. All the children were screened for hearing. The hearing characteristics and long-term follow-up results of homozygous mutations at different gene sites were analyzed by auditory diagnosis and behavioral follow-up from 1 to 3 years. Result:Fourteen cases of GJB2 c.109G>A with a homozygous mutation, 11 cases passed the hearing screening, the audiological diagnosis was normal, and the behavior test and follow-up were normal from 1 to 3 years. Hearing screening was not passed in 3 newborns, mild to moderate abnormalities of single or bilateral ears were diagnosed by audiology, 1 000 Hz without positive, and middle ear lesions were diagnosed. Eight cases of GJB2 c.235del C homozygous mutation were followed up by behavioral audiometry and follow-up from 1 to 3 years after cure. Among them, 5 cases were diagnosed as severe hearing impairment of bilateral ears and 3 cases as mild and moderate hearing impairment. One case of GJB3 547G>A homozygous mutation was followed up for 1-3 years, and all of them failed to pass the follow-up of behavioral audiometry and follow-up. Four cases of SLC26A4 IVS7-2A>G, 1 case of SLC26A4 1229C>T homozygous mutation, all of them failed to pass the neonatal hearing screening. All the patients were diagnosed as severe hearing impairment of binaural hearing, and the follow-up of 1-3 years' follow-up did not pass the follow-up tests. Conclusion:GJB2 C.235del C, SLC26A4 IVS7-2A>G locus homozygous mutation infant hearing impairment was mainly severe hearing impairment in bilateral ears, and there was no change in 1-3 years follow-up. GJB2 C.109G A homozygous mutant infants had normal hearing, and it was suggested that they should be followed up closely. It is very important to give correct and reasonable genetic counseling to parents with GJB2 C.109G A homozygous mutation without unnecessary panic.

LINC00324 exerts tumor-promoting functions in lung adenocarcinoma via targeting miR-615-5p/AKT1 axis.

OBJECTIVE: The underlying mechanism of long non-coding RNA (lncRNA) in lung adenocarcinoma (LAC) has not been fully understood yet. Hence, this study aimed to determine the biological function of LINC00324 in LAC and to provide a novel diagnostic and therapeutic target for it. PATIENTS AND METHODS: The expression level of LINC00324 in 87 paired LAC tumor tissues and matched para-tumor tissues was detected using quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Cell counting kit-8 (CCK-8) assay was employed to analyze the cell proliferative ability, whereas flow cytometry was performed to detect cell apoptotic rate. Cell metastasis change was measured using wound-healing assay and transwell assay. Luciferase reporter gene assay and Western blotting analysis were utilized to investigate the underlying mechanism of LINC00324 in LAC. RESULTS: LINC00324 was highly expressed in LAC tissues compared with the para-tumor samples. Identically, the expression level of LINC00324 was significantly higher in LAC cell lines. The overexpression of LINC00324 promoted cell proliferation and inhibited cell apoptosis of LAC cells, while knockdown of LINC00324 presented the opposite effect. Up-regulation of LINC00324 accelerated cell migration and invasion, but down-regulation of LINC0324 decreased cell metastasis of LAC cells. Furthermore, miR-615-5p was found to be regulated by LINC00324 and inhibited AKT1 expression, indicating that LINC00324 promoted cell progression via affecting the miR-615-5p/AKT1 pathway. CONCLUSIONS: LINC00324 was significantly over-expressed in LAC tissues and cells. It promoted proliferation and metastasis but inhibited cell apoptosis of LAC cells via sponging miR-615-5p to promote AKT1 expression. Our results demonstrated LINC00324 as a novel diagnostic and therapeutic target for LAC.

[Efficacy of Yisaipu tapering in the treatment of ankylosing spondylitis].

Objective: To assess the efficacy of Yisaipu tapering in patients with ankylosing spondylitis (AS). Methods: A total of 87 cases of AS patients from Guangdong Second Provincial General Hospital who were treated with Yisaipu and celecoxib were retrospectively analyzed from February 2013 to April 2017.All patients received full dose Yisaipu and celecoxib in the initial 12 weeks.After that, the patients in the full dose group maintained Yisaipu (50 mg/w) treatment from the 13(rd) to 24(th) week, while tapering group received Yisaipu 50 mg subcutaneous injection once every other week.By using AS disease activity score (ASDAS), Bath AS functional index (BASFI) and magnetic resonance (MR) score of sacroiliac joint (SIJ) plus recording adverse events, differences of efficacy and safety between groups were compared. Results: ASDAS and BASFI of tapering group were 1.1±0.7 and 1.3±1.1, while those of full dose group were 1.0±0.7 and 1.1±1.0, respectively.No significant difference of ASDAS or BASFI was found between groups.Besides, the MR scores of tapering and full dose groups were 8±7 and 8±6 respectively before therapy, while they were significantly lower in the 24(th) week (4±4 and 4±3, P<0.05). However, changes of MR score between groups were similar (P>0.05). Conclusion: Dose tapering of Yisaipu subcutaneous injection might be effective for keeping stable of disease activity and function in patients with AS.Its efficacy is similar to those of full dose Yisaipu.

Functional and mutational analyses of an omega-class glutathione S-transferase (GSTO2) that is required for reducing oxidative damage in Apis cerana cerana.

Glutathione S-transferases perform a variety of vital functions, particularly in reducing oxidative damage. Here, we investigated the expression patterns of Apis cerana cerana omega-class glutathione S-transferase 2 (AccGSTO2) under various stresses and explored its connection with antioxidant defences. We found that AccGSTO2 knockdown by RNA interference triggered increased mortality in Ap. cerana cerana, and immunohistochemistry revealed significantly decreased AccGSTO2 expression, particularly in the midgut and fat body. Further analyses indicated that AccGSTO2 knockdown resulted in decreases in catalase and glutathione reductase activities, ascorbate content and the ratio of reduced to oxidized glutathione, and increases in H2 O2 , malondialdehyde and carbonyl contents. We also analysed the transcripts of other antioxidant genes and found that many genes were down-regulated in the AccGSTO2 knockdown samples, revealing that AccGSTO2 may be indispensable for attaining a normal lifespan by enhancing cellular oxidative resistance. In addition, the roles of cysteine residues in AccGSTO2 were explored using site-directed mutagenesis. Mutants of Cys(28) and Cys(124) significantly affected the enzyme and antioxidant activities of AccGSTO2, which may be attributed to the changes in the spatial structures of mutants as determined by homology modelling. In summary, these observations provide novel insight into the structural and functional characteristics of GSTOs.

Potential hippocampal genes and pathways involved in Alzheimer's disease: a bioinformatic analysis.

Alzheimer's disease (AD) is a neurodegenerative disor-der and the most common cause of dementia in elderly people. Nu-merous studies have focused on the dysregulated genes in AD, but the pathogenesis is still unknown. In this study, we explored critical hippocampal genes and pathways that might potentially be involved in the pathogenesis of AD. Four transcriptome datasets for the hip-pocampus of patients with AD were downloaded from ArrayExpress, and the gene signature was identified by integrated analysis of mul-tiple transcriptomes using novel genome-wide relative significance and genome-wide global significance models. A protein-protein interaction network was constructed, and five clusters were selected. The biologi-cal functions and pathways were identified by Gene Ontology and Kyo-to Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. A total of 6994 genes were screened, and the top 300 genes were subjected to further analysis. Four significant KEGG pathways were identified, including oxidative phosphorylation and Parkinson's disease, Huntington's disease, and Alzheimer's disease pathways. The hub network of cluster 1 with the highest average rank value was de-fined. The genes (NDUFB3, NDUFA9, NDUFV1, NDUFV2, NDUFS3, NDUFA10, COX7B, and UQCR1) were considered critical with high degree in cluster 1 as well as being shared by the four significant path-ways. The oxidative phosphorylation process was also involved in the other three pathways and is considered to be relevant to energy-related AD pathology in the hippocampus. This research provides a perspec-tive from which to explore critical genes and pathways for potential AD therapies.

14-3-3 gene expression in regenerating rat liver after 2/3 partial hepatectomy.

14-3-3 Proteins are a ubiquitous family of molecules that participate in protein kinase signaling pathways in all eukaryotic cells. Functioning as phosphoserine/phosphothreonine-binding modules, 14-3-3 proteins participate in the phosphorylation-dependent protein-protein interactions that control progression through the cell cycle, initiation and maintenance of DNA damage checkpoints, activation of MAP kinases, prevention of apoptosis, and coordination of integrin signaling and cytoskeletal dynamics. During liver regeneration after partial hepatectomy, normally quiescent hepatocytes undergo hypertrophy and proliferation to restore the liver mass. In this study, we investigated the expression patterns of 14-3-3 mRNAs in regenerating rat liver after 2/3 partial hepatectomy using real-time quantitative reverse transcription-polymerase chain reaction. All mRNAs of the 14-3-3 7 isotypes were expressed at 10 time points. Upregulation of 14-3-3x mRNA expression and downregulation of 14-3-3s mRNA expression from 0 to 6 h may play important roles in the entry into S-phase. Downregulation of 14-3-3b, g, s, h, and t mRNA expression from 24 to 30 h, when compared to 0 h, was closely related to entry into mitosis.

Identification of a novel NPR1-like gene from Nicotiana glutinosa and its role in resistance to fungal, bacterial and viral pathogens.

The NPR1 or NPR1-like genes play a pivotal role in systemic acquired resistance in plants. Here, we isolated and identified a novel tobacco (Nicotiana glutinosa) NPR1-like gene (designated as NgNPR3). The full-length cDNA is 2049 bp in length with a 1767 bp open reading frame which encodes a 588 amino acids protein with an estimated molecular mass of 66 kDa and a calculated pI of 7.14. Homology analysis suggested that the NgNPR3 protein shares significant similarity to AtNPR3 of Arabidopsis. Transient expression assay of NgNPR3-GFP fusion gene in onion epidermal cells revealed that the NgNPR3 protein was localized to the cytoplasm and moved into the nucleus after redox change. RT-PCR results indicated that NgNPR3 was up-regulated after treatment with SA, INA, H(2)O(2,) and MeJA, which play important roles in various resistance responses in tobacco. Transcriptional level of NgNPR3 was also up-regulated after inoculation with Rhizoctonia solani, Phytophthora parasitica, Alternaria alternata, Pseudomonas solanacearum, and potato virus Y (PVY), respectively. When NgNPR3 was overexpressed in N. tabacum cv. Samsun plants, the transgenic plants showed enhanced resistance to the pathogens A. alternate, P. solanacearum and PVY. Furthermore, NgNPR3-mediated disease resistance is dosage-dependent. Our results suggest that NgNPR3 could be a putative NPR1-like gene, and might play an important role in resistance to a broad range of pathogens in tobacco.

Human primary corneal fibroblasts synthesize and deposit proteoglycans in long-term 3-D cultures.

Our goal was to develop a 3-D multi-cellular construct using primary human corneal fibroblasts cultured on a disorganized collagen substrate in a scaffold-free environment and to use it to determine the regulation of proteoglycans over an extended period of time (11 weeks). Electron micrographs revealed multi-layered constructs with cells present in between alternating parallel and perpendicular arrays of fibrils. Type I collagen increased 2-4-fold. Stromal proteoglycans including lumican, syndecan4, decorin, biglycan, mimecan, and perlecan were expressed. The presence of glycosaminoglycan chains was demonstrated for a subset of the core proteins (lumican, biglycan, and decorin) using lyase digestion. Cuprolinic blue-stained cultures showed that sulfated proteoglycans were present throughout the construct and most prominent in its mid-region. The size of the Cuprolinic-positive filaments resembled those previously reported in a human corneal stroma. Under the current culture conditions, the cells mimic a development or nonfibrotic repair phenotype.

DNA methylation and loss of protein expression in esophageal squamous cell carcinogenesis of high-risk area.

DNA methylation is an important mechanism for gene silence. The purpose of this study was to investigate aberrant promoter methylation of the p16 and FHIT genes in tissues and plasma and loss of protein expression in esophageal precancerous conditions (EPC) and esophageal squamous cell carcinoma (ESCC) of high-risk area. Methylation-specific PCR(MSP) was employed to examine the DNA methylation in the plasma and tissues of 95 patients of EPC, ESCC and 10 chronic esophagitis (CE). Loss of protein expression of p16 and FHIT was detected immunohistochemically. In total 95 lesion tissues of EPC and ESCC, p16 methylation was found in 53 (55.79%) cases, and 41 of 53 (77.36%) cases were demonstrated deletion of the p16 protein immunohistochemically. FHIT methylation was found in 49 (51.58%) cases, and 40 of 49 (81.63%) were demonstrated deletion of the FHIT protein. Only 1 (10%) case of 10 CE p16 methylation was found in the tissues. In the plasma of total 105 samples, 2 of 23 high grade intraepithelial neoplasia (HGIN) and 12 of 37 ESCC were detected p16 methylation, and 2 of 23 HGIN and 14 of 37 ESCC were detected FHIT methylation. These results indicate that p16 and FHIT methylation may be one of the earliest events and an important mechanism for gene silencing in esophageal squamous cell carcinogenesis. This study may be helpful for screening the candidate molecular markers for early diagnosis of ESCC in high-risk area.