[Post-transcriptional regulation mechanism and antiviral strategy of hepatitis B virus RNA].

Chronic hepatitis B virus (HBV) infection is one of the major public health issues of ongoing global concern. Due to inadequate understanding of the HBV life cycle, there is a lack of effective drugs to cure chronic hepatitis B. During HBV replication, covalently closed circular DNA (cccDNA) serves as the template for viral replication and can be transcribed to produce five viral RNAs of 3.5, 2.4, 2.1 kb and 0.7 kb in length, which are translated to produce HBeAg, core protein, polymerase (P) protein, HBsAg and HBx proteins, respectively. Among them, the 3.5 kb pregenomic RNA (pgRNA) is also the template for viral reverse transcription. Polymerase protein recognizes and binds to the capsid assembly signal on the pgRNA to initiate capsid assembly and reverse transcription. Recent studies have revealed that the processes of splicing, nuclear export, stability, translation, and pgRNA encapsidation of HBV RNAs are regulated by a post-transcriptional regulatory network within the host cell and depend on unique post-transcriptional regulatory elements in the HBV RNA structure. The aim of this review is to overview the post-transcriptional regulatory mechanisms of HBV RNA and their applications in the study of HBV antiviral therapeutics, with the aim of providing new ideas for the development of new drugs targeting HBV RNA.

[Effects of postoperative urine protein levels on predicting acute kidney injury in critically ill patients undergoing non-cardiac surgery].

Objective: To investigate the predictive value of postoperative urine protein level in critically ill patients undergoing non-cardiac surgery with acute kidney injury (AKI). Methods: A total of 661 critically ill patients undergoing non-cardiac surgery, who visited the Department of Critical Care Medicine of Peking University First Hospital from May 20, 2019 to November 24, 2020, were enrolled in this prospective study. The clinical data of the patient's age, gender, body mass index, laboratory examination, surgical status, etc. were collected. AKI diagnostic criteria of the 2012 KDIGO guidelines were used to diagnose the occurrence of AKI after surgery. The independent predictors of AKI were determined by multivariate logistic regression. Results: The age of this patient cohort was (69±15) years. The prevalence of AKI was 45.4% (300/661). Multivariate logistic regression showed that urine protein semi-quantitative ≥2+(OR=2.62, 95%CI: 1.05-6.56, P=0.039) was independent factor for postoperative AKI in critically ill patients undergoing non-cardiac surgery, other independent factors include higher age (OR=1.04, 95%CI: 1.02-1.06, P=0.001), higher body mass index (BMI) (OR=1.12, 95%CI: 1.04-1.21, P=0.004), lower plasma hemoglobin level (OR=0.98, 95%CI: 0.97-1.00, P=0.019), lower central venous pressure (OR=0.89, 95%CI: 0.83-0.97, P=0.005) and lower total hypotension time (OR=1.01, 95%CI: 1.00-1.01, P=0.041). Conclusions: Urine protein semi-quantitative ≥2+after surgery is an independent predictive factor for the occurrence of postoperative AKI in critically ill patients undergoing non-cardiac surgery. It is important to check urine routine immediately after surgery to detect and deal with high-risk patients.

A long non-coding RNA, ELFN1-AS1, sponges miR-1250 to upregulate MTA1 to promote cell proliferation, migration and invasion, and induce apoptosis in colorectal cancer.

OBJECTIVE: Long non-coding RNA (lncRNA), is essential for the development and progression of cancers. LncRNA regulates target gene expression by sponging the corresponding microRNA (miRNA) during tumorigenesis. This work aimed to explore the role of one lncRNA, ELFN1-AS1, in colorectal cancer (CRC) development and elucidate the pertinent signaling pathway. PATIENTS AND METHODS: First, we found that ELFN1-AS1 was highly abundant in the human CRC tissues and cell lines. Silence of ELFN1-AS1 expression reduced cell proliferation, colony formation, migration and invasion, while inducing apoptosis in vitro; moreover, knockdown of ELFN1-AS1 decreased the size and weight of tumor in vivo. RESULTS: Luciferase reporter assay revealed that ELFN1-AS1 interacted with miR-1205 and suppressed its expression. In addition, miR-1205 could bind to the 3' untranslated region (3'-UTR) of Metastasis Associated Protein1 (MTA1) and inhibited ELFN1-AS1 expression. More importantly, overexpression of MTA1 completely rescued the phenotype of ELFN1-AS1 knockdown. CONCLUSIONS: In sum, our study demonstrated that ELFN1-AS1 sponges miR-1205 to upregulate MTA1, which is essential for CRC cell proliferation, migration, and invasion as well as apoptosis induction.

LncRNA AOC4P affects biological behavior of gastric cancer cells through MAPK signaling pathway.

OBJECTIVE: Long non-coding RNA (lncRNA) is closely related to the occurrence and development of gastric cancer, but the mechanism and clinical significance of lncRNA AOC4P are still unclear. This study aimed to investigate the expression and function of lncRNA AOC4P in gastric cancer. PATIENTS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was used to detect the expression of lncRNA AOC4P in 80 gastric cancer tissues and adjacent normal tissues. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide), flow cytometry and transwell assays were used to study the effects of lncRNA AOC4P on the proliferation, apoptosis, migration and invasion of gastric cancer cells. Western blot was used to detect the related protein level of the mitogen-activated protein kinase (MAPK) signal pathway. RESULTS: The expression of lncRNA AOC4P in gastric cancer tissues was higher than that in adjacent tissues. OS or DFS time were significantly shortened in patients with gastric cancer with high expression of lncRNA AOC4P. Inhibition of lncRNA AOC4P expression can inhibit cell proliferation, migration and invasion, promoting cell apoptosis to some extent. Inhibition of lncRNA AOC4P expression also can result in the decreased expression levels of extracellular-signal-regulated kinase 1 (ERK1), c-Jun N-terminal kinases (JNK) and p38 proteins. CONCLUSIONS: High expression of lncRNA AOC4P in gastric cancer may be related to the occurrence, development and prognosis of gastric cancer. LncRNA AOC4P is expected to become a new diagnostic marker and therapeutic target for gastric cancer.

LncRNA CASC19 promotes the proliferation, migration and invasion of non-small cell lung carcinoma via regulating miRNA-130b-3p.

OBJECTIVE: To uncover the biological role of long non-coding RNA (lncRNA) CASC19 in the pathogenesis of non-small cell lung carcinoma (NSCLC) and the potential mechanism. PATIENTS AND METHODS: Expression pattern of lncRNA CASC19 in NSCLC tissues and cell lines was determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Survival analysis on the correlation between CASC19 level and prognosis of NSCLC patients was conducted by introducing for the Kaplan-Meier estimator. After the transfection of si-CASC19 in A549 and PC9 cells, changes in viability, migratory, and invasive capacities were evaluated. Dual-luciferase reporter gene assay was performed to explore the interaction between microRNA-130b-3p (miRNA-130b-3p) and CASC19/ZEB2. Their interactive effects on the progression of NSCLC were finally investigated through rescue experiments. RESULTS: LncRNA CASC19 was upregulated in NSCLC tissues and cell lines. NSCLC patients with high expression of CASC19 presented a worse survival. Knockdown of CASC19 attenuated proliferative, migratory, and invasive capacities of A549 and PC9 cells. CASC19 sponged miRNA-130b-3p and negatively regulated its level. ZEB2 was the direct target of miRNA-130b-3p. The knockdown of miRNA-130b-3p reversed the regulatory effects of CASC19 on A549 and PC9 cells. CONCLUSIONS: CASC19 sponges miRNA-130b-3p to regulate ZBR2 as a ceRNA, thus accelerating the progression of NSCLC by regulating proliferative, migratory, and invasive capacities of tumor cells.

[Molecular analysis of gene mutations in eight patients with Glanzmann's thrombasthenia].

Objective: To analyze the gene sequencing in eight patients with Glanzmann's thromboasthenia(GT), and combined with clinical manifestations and laboratory findings to investigate the molecular mechanism of GT. Methods: Eight patients who were diagnosed as GT based on platelet aggregation test and flow cytometry were enrolled, as well as 4 pedigrees. Next-generation sequencing was used to analyze all the exons and flanking sequences of αⅡ band ß3 gene and also platelet-type bleeding disorders related genes. Gene polymorphism was excluded by retrievaling HGMD and PubMed databases and relative literature. Mutations were confirmed by sanger sequencing. Results: All the eight patients had relatively normal platelet counts and coagulation profiles. But their platelet response to ADP was impaired, and their platelet response to ristocetin was relatively normal. Flow cytometry showed that of the 8 patients, platelet surface αⅡb/ß3 was lower than 5% of the normal value in 5 cases, and in 2 cases was 5% to 20% of normal value, and in 1 case there was no significant platelet surface αⅡb/ß3 reduction compared with normal level. Gene analysis revealed that five mutations in ITGA2B gene were identified, including c. 1750C>T(p.Arg584Ter), c.1882C>T(p.Arg628Ter), c.814G>C(p.Val272Leu), c.2333A>C(p.Gln778Pro), c.432G>A(p.Trp144Ter). Six mutations in ITGB3 gene, including c. 719G>A(p.Arg240Gln), c.2248C>T(p.Arg750Ter), c.1495T>C(p.Cys499Arg), c.1728delC(p.Ser577ProfsTer92), c.877C>T(p.Gln293Ter), c. 1260G>A were identified. In addition, mutations in genes such as RUNX1, HPS4, MYH9, ACTN1, HPS3 and SETBP1 were identified in patients with GT. Conclusions: Rather than homozygous mutations, heterozygous mutations, especially compound heterozygous mutations, are more common in patients with GT. The pathogenesis of GT may relate to gene mutations such as RUNX1 in addition to the ITGA2B gene and the ITGB3 gene.

Current views of pediatric asthma.

Childhood asthma is influenced by multiple factors including genetic, socioeconomic, socio demographic and environmental factors. The symptoms of childhood asthma are observed to be variable. Some studies reported that asthma prevalence is disproportionately high among socially disadvantaged children. On the other hand, some reports found weak or no association between social disadvantage and childhood asthma. Recent literature showed that growth of health-related quality of life (HRQOL) instruments in the management of childhood asthma. The present review article would discuss the current views and the latest developments in the field of pediatric asthma.

[Clinical Application Value of Peripheral Blood Diagnostic Report].

Objective: To explore the clinical application value of peripheral blood diagnostic report. Methods: 557 peripheral blood diagnostic reports were collected from Peking University First Hospital, YANDA LU DAOPEI Hospital and Beijing United Family Hospital. The results were analyzed and summarized according to different blood cell morphology character for the first time and review cases, respectively. Results: Two hundred and one samples from first time patients were found abnormal complete blood count or leukocyte differential count, they were summarized as anemia, anemia accompanied with leukopenia or thrombopenia, abnormal white blood cell count or leukocyte differential count and abnormal platelet count. Each condition was further distinguished on the basis of different morphology character. Initial diagnosis or further examination could be proposed if abnormal morphology was specific or typical, when blood cell morphology was atypical or normal, the morphology was described objectively. 22 review cases included many benign and malignant disorders such as acute leukemia, chronic leukemia, myelodysplastic syndrome, multiple myeloma, infectious mononucleosis and so on. Suggestion of therapeutic effect, progression of diseases or further examination could be present according to complete blood cell count and morphology character. Conclusion: Peripheral blood diagnostic report can provide more comprehensive and accurate information for clinic, and propose important advisory opinions for primary diagnosis, differential diagnosis, treatment monitoring and progression assessment.

[The preparation and evaluation of the quality control materials for detection of platelet membrane glycoproteins by flow cytometry].

Objective: To prepare the quality control material for detection of platelet membrane glycoproteins by flowcytometry and evaluate the appearance traits, homogeneity and stability of it. Methods: Fresh platelets from the blood group O donors were fixed by the certain concentration of aldehyde solution and then washed by the imidazole buffer. After that, adding certain concentration of lyophilized protection solution into the preparations. The preparations were dispensed to be lyophilized and then were kept refrigerated in 2-8 ℃.According to the protocol of control of lyophilized biological products, the quality indicator for monitoring the prepared process, containing the appearance traits, the residual water, the platelet recovery and the rehydration quality were evaluated. The homogeneity and stability of these preparations were evaluated according to the CNAS-GL03 Guidance on evaluating the homogeneity and stability of samples used for proficiency testing and the ISO Guide 35 Reference material-general and statistical principles for certification. Results: The appearance traits and the rehydration quality of the quality control materials meeted the requirements, with the residual water distributed between 3.96% to 4.04% and the platelet recovery rate ranged from 68% to 72%.The homogeneity evaluation showed that there was no significant difference among the groups(P>0.05). The stability test indicated that the positive rate of platelet membrane glycoproteins CD42b, CD41 and CD62P of the quality control material was -0.14%, -0.14% and 0.74%, respectively, at 16 weeks after storage. There was no linear trend between the percentage of positive platelets with membrane glycoproteins and time(P>0.05). Conclusions: The quality control material for detection of platelet membrane glycoproteins by flow cytometry prepared by us meets the needs of the appearance traits, the residual water, the rehydration quality, the homogeneity and the longtime stability.It is hopeful to be used as internal quality control of the assay in clinic laboratory, the external quality assessment and proficiency evaluation.

[Basic characteristics and survival analysis of patients with hepatocellular carcinoma].

Objective: To analyze the basic characteristics of patients with hepatocellular carcinoma (HCC), and further explore the major factors affecting the prognosis of HCC patients. Methods: A total of 800 HCC patients were randomly selected from the Cancer Hospital, Chinese Academy of Medical Sciences. Their clinical and follow-up information was obtained from medical record. Univariate analysis of variance, Kaplan-Meier method and Cox regression analysis were used to analyze the patients' age at diagnosis and survival time, etc. Results: The average age of diagnosis was 55.04 years among all the 800 HCC patients, and the sex ratio of male to female was 4.48. The infection rates of HBV and HCV were 78.6% (629/800) and 5.8% (46/800), respectively. The smoking rate was 41.0% (328/800) and the alcohol consumption rate was 38.5% (328/800). 259 (32.4%) patients underwent radical treatments with liver resection as major therapy, and 541 (67.6%) patients adopted non-radical treatments with transcatheter arterial chemoembolization (TACE) as major therapy. The 1-, 3- and 5-year survival rates of the HCC patients were 73.2%, 53.7% and 42.4%, respectively. The risk factors for prognosis included alcohol abuse and treatment methods. The HR of alcohol abuse was 1.326 (95%CI: 1.058 to 1.661) and HR of treatment methods was 3.301 (95% CI: 2.483 to 4.387). Conclusions: Men account for the majority of HCC patients, and most patients have a lower age at diagnosis and adopt non-radical treatments. The exposure rates of HBV infection and alcohol abuse of HCC patients are significantly higher than those of general population. The major risk factors affecting prognosis and survival are treatments and alcohol abuse. Alcohol abuse and HBV may have synergistic effects on the survival of HCC patients.

A High-sugar High-fat Diet Induced Metabolic Syndrome Shows some Symptoms of Alzheimer's Disease in Rats.

OBJECTIVES: Cases of sporadic Alzheimer's disease (SAD) are the predominant form of the age-related dementia. New evidence suggests that metabolic syndrome (MS), a metabolic disorder, is an initiating factor of some SAD cases. A high-sugar high-fat diet could cause MS, we aimed to investigate whether it could directly lead to SAD. MEASUREMENTS: The characteristic molecules of AD (hippocampus Aß and Tau) were tested by using ELISA and western blotting to confirm the happening hallmarks of AD in brain. MS and inflammation related biochemical indicators were measured using immunological method. Proteins associated with the insulin resistance signal pathway (JNK, PI-3K, AKT, GSK-3ß, GLUT3) were evaluated using western blotting method. The levels of reactive oxygen species (ROS) were measured by immunofluorescence method. RESULTS: Expressions of hippocampus Aß, phosphorylation-Tau (p-Tau), inflammatory factors and p-JNK, Gsk-3ßwere higher in the model rats than those in the control rats and expressions of p-PI3K, p-AKT and GLUT3 were reversed. CONCLUSIONS: The MS model animals, which can induce the characteristics symptoms of AD, and therefore it may be preliminarily considered that the AD pertains to the MS-related diseases.

Association between oral leukoplakia and upper gastrointestinal cancers: a 28-year follow-up study in the Linxian General Population Trial.

BACKGROUND: Oral leukoplakia is a precancerous disorder that is common among residents in Linxian. However, the associations between oral leukoplakia and upper gastrointestinal cancers have not been reported. We investigated the relationships between oral leukoplakia and upper gastrointestinal cancers in the Linxian General Population Trial cohort. METHODS: The Linxian General Population Trial cohort, with 29,584 healthy adults enrolled in 1985 and followed through the end of 2012. With collected baseline data, hazard ratios (HR) and 95% confidence intervals (95% CI) for developing upper gastrointestinal cancers were estimated using Cox proportional hazard models. RESULTS: During 28 years of follow-up, we confirmed a total of 2924 incident esophageal squamous cell carcinoma (ESCC) cases, 1644 gastric cardia cancers and 590 gastric non-cardia cancers. Overall, participants with oral leukoplakia had significantly higher risk of developing ESCC (HR=1.18, 95% CI: 1.08, 1.29). Among individuals ⩽52 years old at baseline, oral leukoplakia was associated with elevated risk of ESCC (HR=1.31, 95%CI: 1.15, 1.49). No significant associations were observed for gastric cardia or non-cardia cancers in either all subjects or subgroups. CONCLUSIONS: Oral leukoplakia was associated with increased risk of ESCC, particularly in younger population. Future studies are needed to confirm these findings.

Body mass, tobacco smoking, alcohol drinking and risk of cancer of the small intestine--a pooled analysis of over 500,000 subjects in the Asia Cohort Consortium.

BACKGROUND: The evidence for a role of tobacco smoking, alcohol drinking, and body mass index (BMI) in the etiology of small intestine cancer is based mainly on case-control studies from Europe and United States. SUBJECTS AND METHODS: We harmonized the data across 12 cohort studies from mainland China, Japan, Korea, Singapore, and Taiwan, comprising over 500,000 subjects followed for an average of 10.6 years. We calculated hazard ratios (HRs) for BMI and (only among men) tobacco smoking and alcohol drinking. RESULTS: A total of 134 incident cases were observed (49 adenocarcinoma, 11 carcinoid, 46 other histologic types, and 28 of unknown histology). There was a statistically non-significant trend toward increased HR in subjects with high BMI [HR for BMI>27.5 kg/m2, compared with 22.6-25.0, 1.50; 95% confidence interval (CI) 0.76-2.96]. No association was suggested for tobacco smoking; men drinking>400 g of ethanol per week had an HR of 1.57 (95% CI 0.66-3.70), compared with abstainers. CONCLUSIONS: Our study supports the hypothesis that elevated BMI may be a risk factor for small intestine cancer. An etiologic role of alcohol drinking was suggested. Our results reinforce the existing evidence that the epidemiology of small intestine cancer resembles that of colorectal cancer.

Phyletic relationships of protein structures based on spatial preference of residues.

A structure-based scoring matrix MDPRE was derived from amino acid spatial preferences in protein structures. Sequence alignment and evolutionary studies by using MDPRE matrix gave similar results as those from ordinary sequence and structure alignments. It is interesting that a matrix derived from structure data solely could give comparable alignment results, strongly indicating the intimate connection between protein sequences and structures. The branch order and length from this approach were close to those obtained by a structure comparison method. Thus, by applying this structure-based matrix, the trees obtained should reflect evolutionary characteristics of protein structure. This approach takes advantage over a direct structure comparison in that (1) only a sequence and MDPRE matrix are needed, making it simple and widely applicable (especially in the absence of 3-dimensional protein structure data); (2) an established algorithm for sequence alignment and tree building could be employed, providing opportunities for direct comparison between matrices from different methodologies. One of the most striking features of this method is its capability to detect protein structure homologies when the sequence identities are low. This was well reflected in the given examples of the alignment of dinucleotide-binding domains.