[Comparison of trends in congenital heart disease mortality from 1990 to 2017 between China and North America].

Objective: To compare trends in congenital heart disease (CHD) mortality between China and North America from 1990 to 2017. Methods: Using the data from the Global Burden Of Disease (GBD) study 2017, we analyzed the related indicators of CHD mortality in China and North America from 1990 to 2017, including standardized mortality, number of deaths, age distribution of death population and age-specific mortality of CHD in each birth cohort. Age-period-cohort model was used to calculate the annual percent change of age-standardized and age-specific mortality rates of CHD (% per year), period effect-adjusted age-specific mortality rates, and the relative risk of death among CHD population at different time periods (2000－2004 as reference period) and different birth cohorts (1970 as reference cohort). Results: In 2017, the age-standardized mortality rates for CHD in China and North America were 2.63/100 000 and 1.13/100 000 respectively, a decrease of 50.4% and 49.4% compared to 1990. Of all deaths from CHD in China, 76.8% were found in children under 5 years, which was higher than that in North America (51.7%). For population under 40 years, the period effect-adjusted age-specific mortality was higher in China (0.46-167.94 per 100 000 person-years) than in North America (0.68-22.47 per 100 000 person-years); whereas for population over 40 years, mortality was lower in China (0.13-0.34/100 000 person years) than in North America (0.43-0.72/100 000 person-years).From 1990 to 2017, CHD mortality in China decreased by 1.95% per year. The annual decrease of mortality ranged from 1.95% to 3.64% per year in population under 45 years, but the mortality showed increasing trends among those over 50 years. In 2015－2019, the relative risk of death from CHD decreased by 31% in China and 24% in North America. For 2015 birth cohort, the relative risk of death decreased by 84% in China and by 64% in North America. Conclusions: In the past 30 years, the risk of death from CHD in China has significantly decreased, and the survival gap with North America is dramatically narrowed. However, mortality is higher among younger populations in China than in North America, and the mortality in the elders shows increasing trends each year in China.

Hypoxia-responsive miR-346 promotes proliferation, migration, and invasion of renal cell carcinoma cells via targeting NDRG2.

Renal cell carcinoma (RCC) is the most common malignant tumor of the kidney. In this study, we investigated the role of miR-346 in RCC cells under hypoxia. OS-RC-2 and 786-O cells were cultured in 1% O2 or normal oxygen. Cell proliferation, migration, and invasion abilities were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, transwell migration, and invasion assays, respectively. Quantitative real-time PCR (qRT-PCR) was performed to detect the expression of miR-346 and N-myc downstream-regulated gene 2 (NDRG2). Then bioinformatics analysis, dual-luciferase reporter assay, and RNA immunoprecipitation were carried out to determine the relationship between miR-346 and NDRG2. The protein expression of NDRG2 was detected by western blot assay. Hypoxia promoted cell proliferation, migration, and invasion in OS-RC-2 and 786-O cells. Meanwhile, we found that miR-346 was upregulated in RCC cells under hypoxia as relative to normoxia. miR-346 deletion could decrease the viability, migration, and invasion abilities of RCC cells under hypoxia. Besides, our data demonstrated that NDRG2 was a target gene of miR-346. The expression of NDRG2 in OS-RC-2 and 786-O cells was lower under hypoxia than under normal oxygen conditions. Moreover, NDRG2 overexpression could inhibit cell proliferation, migration, and invasion in RCC cells under hypoxia. And NDRG2 silencing reversed the inhibitory effects of the miR-346 inhibitor on the viability, migration, and invasion abilities of RCC cells in hypoxia conditions. miR-346 promoted the viability, migration, and invasion of RCC cells under hypoxia by targeting NDRG2.

Analysis of Consistency between Forensic Psychiatry Identification Diagnosis and Clinical Diagnosis of 1 369 Cases.

ABSTRACT: Objective To explore the consistency between identification diagnosis and pre-identification clinical diagnosis of patients with mental disorder undergoing forensic psychiatry identification. Methods The identification data of 1 369 appraised individuals who underwent criminal responsibility identification carried out by the Forensic Institute of Second Affiliated Hospital of Jining Medical University from 2014 to 2017 were collected retrospectively using self-designed investigation data sorting table. A comparative analysis of the mental disorder diagnosis results of expert opinion and past clinical diagnosis results was made. Results Among 1 369 appraised individuals, 964 cases （70.4%） were identified and diagnosed with mental disorder and 405 cases （29.6%） without mental disorder. Among the former, 63.3% （610 cases） were clinically diagnosed, which was higher than 43.2% （175 cases, P<0.05） in the latter. Among the various mental disorders that had been identified and diagnosed, patients with hysteria, stress, and neurosis had the highest proportion of clinical diagnoses （86.7%）, while patients with mental retardation had the lowest proportion of clinical diagnoses （9.6%）. Schizophrenia had the highest overall consistency rate of identification diagnosis and clinical diagnosis （98.4%）, while personality and behavior disorder had the lowest （33.3%）. The overall consistency rate between clinical diagnosis and identification diagnosis of the mental disorder group was 84.1%, and the Kappa value was 0.759. Compared with clinical diagnosis, the consistency rate between inpatient diagnosis and identification diagnosis was higher （85.9%, P<0.05）. Conclusion In forensic psychiatry identification that provides data of past clinical diagnosis and treatment, a high consistency between identification diagnosis and clinical diagnosis of the appraised individual who is identified and diagnosed with mental disorder exists. Clinical diagnosis （especially the inpatient diagnosis） has a relatively good reference value for forensic psychiatry identification.

Global, regional, and national time trends in myocarditis-related mortality, 1990-2019: An age-period-cohort analysis.

OBJECTIVE: The aim of this study was to analyze the tendency of myocarditis mortality in 204 countries and areas during the last three decades and its connection with age, epoch, and birth cohort. MATERIALS AND METHODS: The Global Burden of Disease 2019 Study acquired a cause-specific myocarditis mortality estimate. The net drift, as well as the influence caused by age, period, and birth cohort, were evaluated by the age-period-cohort model. Additionally, we analyzed the tendency in research intensity and international collaboration across countries using 3,983 myocarditis-related publications from four periods during 1990-2019. RESULTS: During the last three decades, 101 of 204 countries and areas experienced an increase (net drifts ≥0.0%) or stagnant declines (≥-0.5%) in the death rate. In particular, increasing death rate was generally discovered in most countries whose Socio-demographic indexes (SDIs) are high and middle-high, such as the United States [net drift=2.11% (95% CI 1.71-2.51)] and Italy [2.65% (1.24-4.08)]. Countries with a higher number of deaths were more active in this field of study, such as the United States (237 publications), China (120), and Italy (73). The United States and Italy, whose total link strengths were 209 and 135, respectively, were more active in international collaborative studies. CONCLUSIONS: Despite the global decrease in myocarditis death rate during the last three decades, negative period and cohort effects and elevated mortality were discovered in numerous countries, especially in those whose SDIs were high, and the age distribution of deaths shifted from adolescent to middle-aged and older populations. We also observed a decline in myocarditis research in some countries with increased mortality.

MicroRNA-129-5p inhibits invasiveness and metastasis of lung cancer cells and tumor angiogenesis via targeting VEGF.

OBJECTIVE: This study was to find out the influence of microRNA-129-5p on proliferative ability, invasiveness, and metastasis of lung tumor cells and tumor angiogenesis. Besides, the effects of microRNA-129-5p on vascular endothelial growth factor (VEGF) level and potential regulatory mechanisms were also what we were interested in. PATIENTS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) assay was performed to detect the microRNA-129-5p level in tumor tissues and paracancerous tissues of 50 patients with LCa, and the interaction between microRNA-129-5p expression and LCa pathological parameters was analyzed. The untreated cell group (NC) and the transfected microRNA-129-5p overexpression group (microRNA-129-5p mimics) were established, and then, the transfection efficiency of microRNA-129-5p was further verified by qRT-PCR. In H1299 and SPC-A1, cell counting kit-8 (CCK-8), Tube-formation experiments, and transwell invasion and migration tests were performed to evaluate the influence of the microRNA on the biological function of LCa cells. Finally, the potential mechanism of action of VEGF, a downstream gene of microRNA-129-5p, was explored by bioinformatics analysis and recovery experiments. RESULTS: QRT-PCR results showed that the level of microRNA-129-5p in cancer tissues of LCa patients was notably lower than that in normal tissues, and the difference was statistically significant. Compared with patients with highly expressed microRNA-129-5p, patients with low level had higher rates of lymph node or distant metastasis, and the overall survival rate was lower. Compared with NC group, cell proliferation, invasiveness and migration ability, and tumor angiogenesis capacity were strikingly decreased in microRNA-129-5p mimics group. Subsequently, VEGF expression was validated conspicuously enhanced in LCa cell line and tissue and was negatively correlated with microRNA-129-5p. In addition, the recovery experiment demonstrated that overexpression of VEGF could counteract the impact of microRNA-129-5p mimics on tumor angiogenesis and the invasive and migratory capacity of LCa cells, which then together led to the malignant progression of LCa. CONCLUSIONS: The above studies demonstrated that microRNA-129-5p was strikingly correlated with LCa lymph node or distant metastasis and poor prognosis, and it can inhibit the malignant progression of this cancer. The investigation also demonstrated that microRNA-129-5p may inhibit proliferation capacity and invasiveness of LCa cells and tumor angiogenesis via regulating VEGF.

The study of microanatomy of intestinal epithelium in the Chinese soft-shelled turtle (Pelodiscus sinensis).

The microanatomy of the intestinal epithelium in the Chinese soft-shelled turtle (CST) was studied by light and transmission electron microscopy (TEM). The small intestinal epithelium (SIE) was single layered or pseudostratified. The enterocytes contained mitochondria or mitochondria and lipid droplets. The enterocytes were arranged tightly in the apical parts of epithelium and connected by desmosomes and interdigitations. The large intestinal epithelium (LIE) was pseudostratified and the enterocytes did not contain lipid droplets. Enterocytes were arranged compactly in the apical part, forming spaces in the middle and basal parts of epithelium. Numerous mucous cells were scattered in the epithelium and there were intraepithelial lymphocytes (IELs) with their pseudopodia extended into the intestinal lumen. This study provides detailed features of intestinal epithelium in the Pelodiscus sinensis that could be related to function. In addition, these findings are discussed in relation to other vertebrates.

Berbamine inhibited the growth of prostate cancer cells in vivo and in vitro via triggering intrinsic pathway of apoptosis.

BACKGROUND: Berbamine (BBM) has been reported with antitumor activities. BBM inhibited the growth of prostate cancer (PCa) cells and caused vacuolization of mitochondria in preliminary study. We hypothesized BBM could enhance apoptosis of PCa cells through mitochondrial pathway. METHODS: Growth of PC-3 and LNCaP cells treated by BBM was determined by cell viability assay. The morphology of mitochondria was observed by electron microscopy. Apoptosis was quantified by flow cytometry. Expressions of bax/bcl-2, active caspase-9 and active caspase-3 were examined by western blot and real-time PCR. Methazolamide, an inhibitor of cytochrome c, was added to examine its blockade effect on BBM. PC-3 cells were injected subcutaneously into athymic mice, and BBM or saline was administrated intravenously. Diameters of induced tumors were compared, and ratios of apoptotic cells in tumor tissues were calculated. RESULTS: BBM inhibited viability of cultured PC-3 and LNCaP cells in dose- and time-dependent manners. Flow cytometry showed BBM induced apoptosis in cultured cells. Mitochondria showed swelling, vacuolization and fused mitochondrial cristae. Western blot and real-time PCR analysis both showed increases of bax/bcl-2, active caspase-9 and active caspase-3. Methazolamide impeded effect of BBM on inducing apoptosis of cultured cells, and activations of caspase-9 and caspase-3 were also inhibited. Growth of tumors by grafted PC-3 cells was significantly slower in BBM group. Ratios of apoptotic cells in tumors were significantly higher in BBM group. Expressions of active caspase-9 and active caspase-3 in tumors were significantly upregulated by BBM. CONCLUSIONS: BBM exhibited strong anti-PCa activities in vitro and in vivo relying on activating caspase-3 via intrinsic pathway of apoptosis, holding a great promise to develop new strategies for PCa.

Purification, cDNA cloning and northern blot analysis of trehalase of pupal midgut of the silkworm, Bombyx mori.

Trehalase (alpha-glucoside-1-glucohydrolase, EC 3.2.1.28) was purified from silkworm pupal midgut to homogeneity by DEAE-Sepharose CL-6B and hydroxyapatite chromatography, and native gel electrophoresis. The enzyme had a molecular mass of 70 kDa. The N-terminal amino-acid sequence of the intact trehalase and its three fragments by V8 proteinase digestion was determined. Based on the amino-acid sequence, degenerate oligonucleotides were synthesized and used as primers in a polymerase chain reaction (PCR). Using a 0.8 kb PCR product as a hybridization probe, trehalase clones were isolated from the pupal midgut cDNA library. Sequence analysis revealed that the isolated trehalase cDNA contains 3103 nucleotides and comprises 579 amino acids, including a cleavable signal sequence and five potential N-glycosylation sites. Northern blot analysis clearly showed a 3.0 kb transcript in midgut, and Malpighian tubule, but not in fat body, silk gland, ovary, trachea, brain and suboesophageal ganglion.

Molecular characterization of ovary trehalase of the silkworm, Bombyx mori and its transcriptional activation by diapause hormone.

We have isolated a cDNA encoding ovary trehalase of the silkworm, Bombyx mori. Sequence analyses revealed that the isolated cDNA contains 3143 nucleotides and comprises 579 amino acids, including a cleavable signal sequence and five potential N-glycosylation sites. Northern blot analysis showed a 3.0 kb transcript in developing ovaries carrying membrane-bound trehalase. A single copy of trehalase gene was present in the haploid genome of the silkworm. The effect of diapause hormone on the accumulation of trehalase mRNA was examined on developing ovaries in in vivo and in vitro conditions. The synthetic diapause hormone brought about a 6-fold increase in trehalase mRNA content in ovaries 4 h after injection. The similar increase was found in ovaries which were incubated in vitro with diapause hormone. Coincubation of ovaries with diapause hormone and actinomycin D could not increase the mRNA level in ovaries, and maintained a basal level which was found in ovaries incubated without diapause hormone. These results indicate that diapause hormone stimulates transcription of the trehalase gene in developing ovaries of the silkworm.

Differentiation within the genus Leptocarabus (excl. L. kurilensis) in the Japanese Islands as deduced from mitochondrial ND5 gene sequences (Coleoptera, Carabidae).

The phylogenetic trees have been constructed for the mitochondrial ND5 gene sequences from the Japanese Leptocarabus ground beetles, which contain 101 specimens collected from nearly the complete distribution ranges of them consisting of five morphological species, i.e., Leptocarabus procerulus, L. kumagaii, L. hiurai, L. kyushuensis and L. arboreus. On the trees, there are recognized two major lineages, each of which is further divided into two or more sublineages. The phylogenetic lines are geographically linked. Two or more species occur in a single lineage, and the same species appear in different lines. We suggest that transformation from one type of morphology to another took place in parallel in various periods of evolution of the Japanese Leptocarabus. From the phylogenetic tree and the dating from the nucleotide substitution rate and the geohistorical data it is inferred that the ancestry of all the Japanese Leptocarabus species was derived from a protoform of L. kyushuensis inhabited the ancient Japan area, followed by separation into two lineages after split of the Japanese Islands from the Eurasian Continent. They then propagated distribution to occupy their own habitat ranges, during which the morphological transformation took place in some lineages.

High-resolution fiber optic temperature sensors using nonlinear spectral curve fitting technique.

A generic new data processing method is developed to accurately calculate the absolute optical path difference of a low-finesse Fabry-Perot cavity from its broadband interference fringes. The method combines Fast Fourier Transformation with nonlinear curve fitting of the entire spectrum. Modular functions of LabVIEW are employed for fast implementation of the data processing algorithm. The advantages of this technique are demonstrated through high performance fiber optic temperature sensors consisting of an infrared superluminescent diode and an infrared spectrometer. A high resolution of 0.01 °C is achieved over a large dynamic range from room temperature to 800 °C, limited only by the silica fiber used for the sensor.

Cloning and expression analysis of takeout/JHBP family genes of silkworm, Bombyx mori.

A Bombyx EST cDNA database was searched using the Drosophila takeout gene and nine cDNAs were obtained. The homology search suggested that these genes are widespread in insects and organize a large gene family, and that they have hydrophobic ligands. A phylogenetic tree indicated that the genes are first divided into two large groups, juvenile hormone binding protein and other protein genes, and the latter group diversified within a short time at an early stage. The expression study of five Bombyx genes indicated that they are expressed in various tissues and are regulated by development and feeding conditions. The Bombyx genes might have roles related to the regulation of metabolism, growth or development related to nutritional conditions.

Trehalase from male accessory gland of an insect, Tenebrio molitor. cDNA sequencing and developmental profile of the gene expression.

A cDNA of alpha alpha-trehalase (EC 3.2.1.28) from a cDNA library of male bean-shaped accessory gland of the mealworm beetle, Tenebrio molitor, has been isolated by the homology screening approach. Sequence analysis of the cDNA (1830 bp) revealed that the cDNA encoded a protein of 555 amino acids with a calculated M(r) of 64457. The deduced amino acid sequence had significant similarities to rabbit small intestine and Escherichia coli trehalases. Northern blotting and semi-quantitative PCR analyses revealed that a trehalase transcript with about 2.0 kb was abundant in bean-shaped accessory glands. In the glands, the amount of trehalase transcript increased from 1 to 2 days after adult ecdysis. These tissue- and stage-specific gene expressions of trehalase corresponded to the tissue- and stage-specificity of trehalase activity.

Origin and diversification of hindwingless Damaster ground beetles within the Japanese islands as deduced from mitochondrial ND5 gene sequences (Coleoptera, Carabidae).

Genealogical trees have been constructed using mitochondrial ND5 gene sequences of 78 Damaster (s. str.) specimens from all over the Japanese Islands. Eight lineages (called races in this paper) have been recognized. The races are tightly linked to geography with sharp distribution boundaries between them. The races and their distribution ranges do not coincide with those of classical morphology. Based on the observed distribution of the mitochondrial DNA haplotypes and the geohistorical data, we propose a diversification scenario of Damaster.

Sympatric convergence of the color pattern in the Chilean Ceroglossus ground beetles inferred from sequence comparisons of the mitochondrial ND5 gene.

The phylogenetic trees have been constructed using the mitochondrial ND5 gene sequences for 66 specimens of the Chilean Ceroglossus ground beetles collected from various localities of Chile. The trees show that the Ceroglossus specimens examined are composed of four phylogenetic lineages that have diverged 25-30 MYA. The results are consistent with the classification proposed by Jiroux (1996) based on morphology, in which Ceroglossus is divided into four species groups. Despite a remarkable color polymorphism revealed by these ground beetles, the color pattern is geographically linked and is not species-specific, suggesting that some ecological or environmental factors are involved in determining it.

Evolutionary discontinuity of the Carabine ground beetles.

The Carabine ground beetles are mostly hind wing-less and cannot fly, so that there is more chance of diversification by geographic isolation compared with winged insects. The relationships between morphological diversification and phylogeny of the ground beetles of the world have been inferred mainly by comparisons of mitochondrial ND5 gene sequences. Based on dating by a mitochondrial DNA "clock," it has been deduced that an explosive radiation of the major carabine groups took place 50-40 MYA. This was followed by occasional radiations on various scales, sometimes accompanied by parallel morphological changes. There are also a good number of examples showing that the fundamental morphology has remained unchanged for a long time among geographically isolated populations within the same species. Thus, carabid evolution would have proceeded discontinuously, with phases of rapid morphological change alternating with silent phases.

Morphological changes in locus ceruleus of albino rats in relation to aging.

Comparative studies were carried out under light, fluorescence- and electron-microscopic observations between young, adult and aged groups of albino rats with the following results: the neuronal packing density of locus ceruleus decreases with advanced age. In addition to the senescent loss of nerve cells, the surviving ones showed some shrinkage of perikarya and reduction or loss of Nissl substance. Some of them presented pyknosis, chromatolysis and loss of RNA in cytoplasm. Glial cells proliferated around the degenerating cells. Senescent cells showed an increase of heterochromatin and decrease of euchromatin. Changes in cytoplasmic organelles included the decrease of ribosomes, alterations in mitochondria and increase of lipofuscins. These senescent changes were thought to affect the absorption of nutrition, message exchange, protein synthesis, supply of energy and transport of materials by the nerve cells.

Phylogenetic relationships and evolution of the Japanese Carabinae ground beetles based on mitochondrial ND5 gene sequences.

The phylogenetic relationships of the Japanese Carabinae ground beetles were analyzed by comparing 1,069 nucleotide sequences in the mitochondrial gene encoding NADH dehydrogenase subunit 5 (ND5). The ND5 phylogenetic tree revealed that the hind-wingless Carabina and the hind-winged Calosoma/Campalita (Calosomina) diverged from the common ancestor, and Cychrus (Cychrini) is the outgroup of them. Five distinct clusters (groups) can be recognized in the Carabina, i.e., CARABUS, HEMICARABUS, LEPTOCARABUS, APOTOMOPTERUS, and PROCRUSTES/DAMASTER. The ancestors of these lineages diverged almost at the same time more than 10 Myr ago. The Carabus cluster includes two subclusters, Carabus and Ohomopterus. Two species of Carabus examined are phylogenetically rather remote, while five species among Ohomopterus are closely related to each other. The results suggest that diversification of Carabus started much earlier than that of Ohomopterus, presumably in the Eurasian continent, and that of Ohomopterus in the Japanese archipelago. The branching order in the LEPTOCARABUS lineage was established, Authenocarabus/Pentacarabus being their outgroup. In the DAMASTER/PROCRUSTES lineage, Procrustes is placed as the outgroup of Damaster, with the branching order of Coptolabrus and Acoptolabrus/Damaster. The diversification of the Damaster subspecies appeared to have occurred in the Japanese archipelago earlier than Ohomopterus, and its phylogeny reflects their geographic distribution in the archipelago rather than the morphological characters.