Quantitative changes in mitochondrial DNA copy number in various tissues of pigs during growth.

Mitochondrial DNA (mtDNA) content is dependent on the energy requirements of tissues. To date, no comprehensive study has been conducted to examine mtDNA copy number variations in pigs. In the current study, quantitative polymerase chain reaction was performed to quantify the mtDNA copy number in 15 pig tissue types at 5 growth stages from embryo to adult. Observable differences in mtDNA content were detected in the tissues, including a 6-fold greater mtDNA content in the heart compared with the lung of 180-day-old samples. mtDNA content in the heart, longissimus dorsi muscle, psoas major muscle, kidney, brain, ovary, and subcutaneous adipose increased with growth. Expression of the replicative mitochondrial helicase (TWINKLE), which regulates mtDNA turnover, was significantly associated with changes in mtDNA copy numbers across tissues during growth (r = 0.33, P = 0.01). We demonstrated that the expression levels of mitochondrial genes were positively correlated with mtDNA copy number.

[Intermediate and long-term outcomes of transcatheter closure of congenital coronary cameral fistulas in 66 children].

Objective: To evaluate the intermediate and long-term outcomes and technical aspects of transcatheter closure (TCC) of coronary cameral fistulas (CCF) in pediatric patients. Methods: This was a case-control study. All pediatric patients with CCF who underwent TCC between January 2005 and December 2019 were retrospectively reviewed. Data was collected from medical records, including demographic characteristics, procedural details, intraoperative and postoperative serious adverse events, follow-up results and prognosis. Patients with serious adverse events and without serious adverse events were compared regarding their clinical features and CCF characteristics. Comparisons between groups were performed with independent sample t test, chi-square test or Fisher exact test. Results: A total of 66 CCF patients (34 boys, 32 girls, 3.9 (1.9, 6.2) years old, 15 (11, 20) kg) underwent attempted TCC. All of the CCF were all medium or large fistulas including 55 proximal fistulas (83%) and 11 distal fistulas (17%). The CCF originated more frequently from the right coronary artery (38 cases (58%)), followed by the left coronary artery (28 cases (42%)). The incidence of coronary artery aneurysms (CAA) was 61% (40/66).Procedural treatment was achieved in 64 patients and procedural success was achieved in 59 patients (92%). Six (9%) serious adverse events occurred in 5 patients during the perioperative period. Acute complications included procedure-related death in one patient and acute myocardial infarction in one patient. Periprocedural complications occurred in 3 patients at one day postoperatively including acute myocardial infarction (2 cases), occluder detachment (1 case), and tricuspid chordae tendinae rupture (1 case). Clinical follow-up data were available in 58 of the 62 patients who underwent initial successful TCC with a follow-up period of 9.3 (6.5, 13.4) years. Ten adverse events occurred in 9 patients including 5 complications consisted of aortic valve perforation (1 case), coronary thrombosis (1 case), progressive aneurysmal dilation after reintervention (1 case), and new-onset tricuspid valve prolapse with significant regurgitation (2 cases) and large residual shunts due to fistula recanalization (5 cases). Therefore, the incidence of intermediate and long-term adverse events was 17% (10/58). During the periprocedural and follow-up period, 16 adverse events occurred in 13 patients, whereas no adverse events occurred in 51 patients. Patients with seriovs adverse events presented with larger proportion of large CCF (11/13 vs. 39% (20/51), P=0.005), giant CAA (10/13 vs.14% (7/51), P=0.030), and higher mean pulmonary artery pressure ((20±9) vs.(16±6) mmHg, 1 mmHg=0.133 kPa, t=2.02, P=0.048) compared to patients without serious adverse events. Conclusions: TCC in CCF children appears to be effective with favorable intermediate and long-term outcomes. Strict indication of TCC is mandatory.

Symmetry-broken Josephson junctions and superconducting diodes in magic-angle twisted bilayer graphene.

The coexistence of gate-tunable superconducting, magnetic and topological orders in magic-angle twisted bilayer graphene provides opportunities for the creation of hybrid Josephson junctions. Here we report the fabrication of gate-defined symmetry-broken Josephson junctions in magic-angle twisted bilayer graphene, where the weak link is gate-tuned close to the correlated insulator state with a moiré filling factor of υ = -2. We observe a phase-shifted and asymmetric Fraunhofer pattern with a pronounced magnetic hysteresis. Our theoretical calculations of the junction weak link-with valley polarization and orbital magnetization-explain most of these unconventional features. The effects persist up to the critical temperature of 3.5 K, with magnetic hysteresis observed below 800 mK. We show how the combination of magnetization and its current-induced magnetization switching allows us to realise a programmable zero-field superconducting diode. Our results represent a major advance towards the creation of future superconducting quantum electronic devices.

Variable selection using the optimal ROC curve: an application to a traditional Chinese medicine study on osteoporosis disease.

In biomedical studies, there are multiple sources of information available of which only a small number of them are associated with the diseases. It is of importance to select and combine these factors that are associated with the disease in order to predict the disease status of a new subject. The receiving operating characteristic (ROC) technique has been widely used in disease classification, and the classification accuracy can be measured with area under the ROC curve (AUC). In this article, we combine recent variable selection methods with AUC methods to optimize diagnostic accuracy of multiple risk factors. We first describe one new and some recent AUC-based methods for effectively combining multiple risk factors for disease classification. We then apply them to analyze the data from a new clinical study, investigating whether a combination of traditional Chinese medicine symptoms and standard Western medicine risk factors can increase discriminative accuracy in diagnosing osteoporosis (OP). Based on the results, we conclude that we can make a better diagnosis of primary OP by combining traditional Chinese medicine symptoms with Western medicine risk factors.

Building a disease risk model of osteoporosis based on traditional Chinese medicine symptoms and western medicine risk factors.

In the Traditional Chinese Medicine (TCM) cross-sectional survey conducted by our team, we were interested in determining the risk factors of osteoporosis. To analyze this TCM study, we had to deal with three statistical problems: (1) a very large number of potential risk factors, (2) interactions among potential risk factors, and (3) nonlinear effects of some continuous-scale risk factors. To address these analytic issues, we used two data mining methods, support vector machine recursive feature elimination and random forest; to deal with the curse of high-dimensional risk factors, we applied another data mining technique of association rule learning to discover the potential associations among risk factors. Finally, we employed the generalized partial linear model (GPLM) to determine nonlinear effects of an important continuous-scale risk factor. The final GPLM model shows that TCM symptoms play an important role in assessing the risk of osteoporosis. The GPLM also reveals a nonlinear effect of the important risk factor, menopause years, which might be missed by the generalized linear model.

Single-cell transcriptomics reveals the heterogeneity of the decidual endothelial cells that participate in labor onset.

OBJECTIVE: To investigate the heterogeneity of decidual endothelial cells and their changes during delivery. PATIENTS AND METHODS: Single-cell RNA sequencing was used to characterize the transcriptomes of decidual endothelial cells before and after the onset of labor. RESULTS: Decidual endothelial cells (9748 cells) were divided into five subgroups with different functions according to differences in the transcriptome. The functions of cluster 5 were enriched in vascular development and response to growth factors. After the onset of labor, the activities of each cluster were different, including the interleukin 17 pathway and regulation of ERK1 and ERK2 cascade. The downregulated genes were related to scavenger receptor (cluster 5), which may reflect the process of endothelial activation. In terms of genetic changes, cluster 5 may be more actively involved in labor than the other clusters. CONCLUSIONS: Peripartum decidual endothelial cells are heterogeneous and participate in labor to varying degrees. One of the five subtypes of endothelial cells may be more actively involved in labor onset. Our findings may enable the assessment of decidual endothelial cells and labor onset.

The appropriate hybrid surgical strategy in three-level cervical degenerative disc disease: a finite element analysis.

OBJECTIVE: The purpose of this FE study was to analyze the biomechanical characteristics of different HS strategies used in the treatment of three-level CDDD (one-level CDA and two-level ACDF). METHODS: We validated the FE model of an intact cervical spine established by transferring the data, collected by 3D CT scan, to the FE software ABAQUS and comparing these data with the data from published studies. Then, the FE model of hybrid surgery was reconstructed to analyze the range of motion (ROM), facet joint force, and stress distribution on an ultrahigh molecular weight polyethylene (UHMWPE) core. RESULTS: The current cervical FE model was able to measure the biomechanical changes in a follow-up hybrid surgery simulation. The total ROM of the cervical HS models was substantially decreased compared with the total ROM of the intact group, and the M2 (C3/4 ACDF, C4/5 CDA, and C5/6 ACDF) model had the closest total ROM to the intact group, but the facet joint force adjacent to the treatment levels showed very little difference among them. The stress distribution showed noticeable similarity: two flanks were observed in the center core, but the inlay of M2 was more vulnerable. CONCLUSIONS: Through the comparison of ROM, the facet joint force after CDA, and the stress distribution of the prosthesis, we find that M2 model has a better theoretical outcome, especially in preserving the maximum total ROM.

In vitro hepatic differentiation of mesenchymal stem cells from human fetal bone marrow.

We examined whether human fetal mesenchymal stem cells (FMSCs) derived from fetal bone marrow were able to differentiate into functional hepatocyte-like cells in vitro. The surface phenotype of FMSCs was characterized by flow cytometry. To induce hepatic differentiation of FMSCs, we added hepatocyte growth factor, basic fibroblast growth factor and oncostatin M into the cell culture medium. After 21 days of hepatocyte induction, FMSCs expressed the hepatocyte-specific markers, alpha-fetoprotein and cytokeratin 18, as demonstrated by immunofluorescence staining. Differentiated FMSCs also demonstrated in vitro functions characteristic of liver cells, including albumin production, urea secretion and glycogen storage. In conclusion, fetal bone marrow-derived FMSCs are able to differentiate into functional hepatocytelike cells and may serve as a source of cells for liver disease therapy.

Inhibiting effect of antisense oligonucleotides phosphorthioate on gene expression of TIMP-1 in rat liver fibrosis.

AIM: To observe the inhibition of antisense oligonucleotides (asON) phosphorthioate to the tissue inhibitors metalloproteinase-1 (TIMP-1) gene and protein expression in the liver tissue of immunologically induced hepatic fibrosis rats. The possibility of reversing hepatic fibrosis through gene therapy was observed. METHODS: Human serum albumin (HSA) was used to attack rats, as hepatic fibrosis model, in which asONs were used to block the gene and protein expressing TIMP-1. According to the analysis of modulator, structure protein, coding series of TIMP-1 genome, we designed four different asONs. These asONs were injected into the hepatic fibrosis models through coccygeal vein. The results was observed by RT-PCR for measuring TIMP-1 mRNA expression, immunohistochemistry and in situ hybridization for collagen I, II, special staining of collagen fiber, and electron microscopic examination. RESULTS: Hepatic fibrosis could last within 363 days in our modified model. The expressing level of TIMP-1 was high during hepatic fibrosis process. It has been proved by the immunohistochemical and the electron microscopic examination that the asON phosphorthioate of TIMP-1 could exactly express in vivo. The effect of colchicine was demonstrated to inhibit the expressing level of mRNA and the content of collagen I, III in the liver of experimental hepatic fibrosis rats. However, the electron microscopy research and the pathologic grading of hepatic fibrosis showed that there was no significant difference between the treatment group and the model group (P> 0.05). CONCLUSION: The experimental rat model of hepatic fibrosis is one of the preferable models to estimate the curative effect of anti-hepatic fibrosis drugs. The asON phosphorthioate of TIMP-1 could block the gene and protein expression of TIMP-1 in the liver of experimental hepatic fibrosis rats at the mRNA level. It is possible to reverse hepatic fibrosis, and it is expected to study a new drug of antihepatic fibrosis on the genetic level. Colchicine has very limited therapeutic effect on hepatic fibrosis, furthermore, its toxicity and side effects are obvious.

Effect of dietary supplemental levels of vitamin A on the egg production and immune responses of heat-stressed laying hens.

Two experiments were conducted to evaluate the effect of vitamin A supplementation of a commercial layer diet on the laying performance and immune function of heat-stressed hens. In Experiment 1, two different levels of vitamin A supplementation (3,000 and 9,000 IU/kg) were used to investigate the laying performance and antibody titer against Newcastle disease virus (NDV) of heat-stressed hens. Results showed that the high level of vitamin A supplementation (9,000 IU/kg) had a beneficial effect on the feed intake and laying rate of heat-stressed hens (P < 0.05), compared with the control group (3,000 IU/kg). The antibody titers were not influenced by the level of vitamin A (P > 0.05). In Experiment 2, the effect of four levels of vitamin A (3,000, 6,000, 9,000, and 12,000 IU/kg) on the antibody titer to NDV and T lymphocyte proportion was studied. The experimental birds were exposed to a high temperature (31.5 C) 15 d after NDV vaccination (Treatment 1) or immediately (Treatment 2). The results showed that the egg weight was increased (P < 0.01) by the high levels of vitamin A supplementation (6,000 and 9,000 IU/kg), but feed intake, laying rate, and body weight loss were not (P > 0.05). In Treatment 1, vitamin A had no significant effect on antibody titers against NDV in normal or hot environments but increased (P < 0.01) the proportion of alpha-naphthyl acetate esterase (ANAE)-positive cells. Vitamin A supplementation had a significant effect on NDV antibody titer and ANAE-positive cell proportion in Treatment 2 (P < 0.01). The results of the present study suggested that vitamin A supplementation in commercial layer diets to layer chickens under heat stress was beneficial to laying performance and immune function.

[Effect of huancongdan capsule on lipoprotein, apolipoprotein and serum immunoglobulin in vascular dementia patients].

OBJECTIVE: To probe into the effect of Huancongdan capsule (HCDC) on the vascular dementia (VD) related factors, including lipoprotein, apolipoprotein (apo) and serum immunoglobulin. METHODS: Fifty-two patients of VD were divided into the HCDC group (27 patients, treated with HCDC) and the control group (25 patients, treated with Naofukang). Changes of related parameters were observed. RESULTS: The effects in lowering triglyceride and total cholesterol and elevating high density lipoprotein cholesterol in the HCDC group were superior to those in the control group (P < 0.01 or P < 0.05), but the effect in lowering low density lipoprotein cholesterol was similar in the two groups (P > 0.05). Compared with data before treatment, in the HCDC group after treatment, the levels of serum apoA1 increased, and serum apoB100, IgA, IgG and circulating immune complex decreased significantly (P < 0.05 or P < 0.01), but the change of apoE level was insignificant (P > 0.05). CONCLUSION: Through adjusting lipid metabolism, HCDC could improve blood rheologic characteristics, promote cerebral blood flow, improve the oxygen carrying capacity of blood to prevent and treat VD. One of its mechanisms is increasing apoA1 and decreasing apoB100 levels, preventing and improving atherosclerosis and reducing damage of vascular endothelial cells, the another may be its action in improving immune function.

[Clinical and laboratory studies of effect of longevity-antihypertensive-mixture on elderly hypertension with kidney deficiency].

UNLABELLED: Longevity-Antihypertensive-Mixture (LAM) is a Chinese herbal drug, which could tonify Kidney in treating the hypertension with Kidney Deficiency in aged patients. RESULTS: 302 patients were randomly divided into three group: The treated group was subdivided into group I (107 Kidney Yin Deficiency patients) and group II (96 non-Kidney Yin Deficiency cases) and group III (99, control group). After treatment of LAM for 8 weeks, the rate of normalizing the blood pressure was 92.52% in group I while in group II, it was 84.43%, the symptoms were also significantly alleviated, and in group III, the rate was 70.71% only. The aorta compliance in group I significantly improved after LAM treatment (P < 0.025), the quality index of left ventricle reduced in treated group, and there was significant difference between treated and control groups. After treatment, LAM could shorten the length of extracorporal thrombus in group. The study showed that LAM could treat hypertension through LAM, administration with good effect.

[Clinical study of bugu shengsui capsule in treating primary osteoporosis with kidney-yang deficiency syndrome].

OBJECTIVE: To assess the effect of Bugu Shengsui capsule (BGSSC) in treating primary osteoporosis. METHODS: Using double energy X-ray bone densimeter and radioimmunoassay (RIA), the effect of BGSSC in treating primary osteoporosis (OP) with Kidney-Yang Deficiency syndrome. RESULTS: The clinical markedly effective and total effective rates of BGSSC were 46% and 82%, respectively. The rate of TCM group is significantly higher than that of the control group (Vitamin D plus calcium). BGSSC could improve bone mineral density by the verification of double energy X-ray bone dansity (DEXA). It could also increase the level of serum calcitonin, luteinizing hormone and calcium, decrease the activity of parathyroid hormone. CONCLUSIONS: The effects of BGSSC on primary OP included increasing patient's bone mineral density, depressing bone resorption and decreasing bone loss.

Human mesenchymal stromal cells ameliorate the phenotype of SOD1-G93A ALS mice.

BACKGROUND: Amyotrophic lateral sclerosis (ALS) is a progressive, lethal, neurodegenerative disease, currently without any effective therapy. Multiple advantages make mesenchymal stromal cells (MSC) a good candidate for cellular therapy in many intractable diseases such as stroke and brain injury. Until now, no irrefutable evidence exists regarding the outcome of MSC transplantation in the mouse model of ALS. The present study was designed to investigate the therapeutic potential of human MSC (hMSC) in the mouse model of ALS (SOD1-G93A mice). METHODS: hMSC were isolated from iliac crest aspirates from healthy donors and kept in cell cultures. hMSC of the fifth passage were delivered intravenously into irradiated pre-symptomatic SOD1-G93A mice. Therapeutic effects were analyzed by survival analysis, rotarod test, motor neuron count in spinal cord and electrophysiology. The engraftment and in vivo differentiation of hMSC were examined in the brain and spinal cord of hMSC-transplanted mice. RESULTS: After intravenous injection into irradiated pre-symptomatic SOD1-G93A mice, hMSC survived more than 20 weeks in recipient mice, migrated into the parenchyma of brain and spinal cord and showed neuroglia differentiation. Moreover, hMSC-transplanted mice showed significantly delayed disease onset (14 days), increased lifespan (18 days) and delayed disease progression compared with untreated mice. DISCUSSION: Our data document the positive effects of hMSC transplantation in the mouse model of ALS. It may signify the potential use of hMSC in treatment of ALS.

Photorealistic virtual anatomy based on Chinese Visible Human data.

Virtual reality based learning of human anatomy is feasible when a database of 3D organ models is available for the learner to explore, visualize, and dissect in virtual space interactively. In this article, we present our latest work on photorealistic virtual anatomy applications based on the Chinese Visible Human (CVH) data. We have focused on the development of state-of-the-art virtual environments that feature interactive photo-realistic visualization and dissection of virtual anatomical models constructed from ultra-high resolution CVH datasets. We also outline our latest progress in applying these highly accurate virtual and functional organ models to generate realistic look and feel to advanced surgical simulators.

Role of fascin in the proliferation and invasiveness of esophageal carcinoma cells.

Fascin, an actin-bundling protein, induces membrane protrusions and increases cell motility in various transformed cells. The overexpression of fascin in esophageal squamous cell carcinoma (ESCC) has been described only recently, but the roles and mechanism still remained unclear. Here, by using RNA interference (RNAi), we have stably silenced the expression of the fascin in EC109 cells, an ESCC cell line. Down-regulation of fascin resulted in a suppression of cell proliferation and as well as a decrease in cell invasiveness. Furthermore, we revealed that fascin might have functions in regulating tumor growth in vivo. The effect of fascin on cell invasiveness correlated with the activation of matrix metalloproteases such as MMP-2 and MMP-9. We examined that fascin down-expression also led to a decrease of c-erbB-2 and beta-catenin at the protein level. These results suggested that fascin might play crucial roles in regulating neoplasm progression of ESCC.

Optical folding-flash analog-to-digital converter with analog encoding.

We describe an optically assisted folding-flash analog-to-digital converter. The periodic transfer function of the Mach-Zehnder interferometer is used to perform analog folding on the electronic signal to be quantized. A novel analog encoding scheme for efficient generation of gray code digital data is proposed. The new encoding scheme eliminates the requirement for interferometers with ultralow V(pi), which, so far, has hindered the development of such systems. The encoding concept is experimentally demonstrated through the use of LiNbO(3) modulators.

The internal methionine codons of human T-cell leukemia virus type II rex gene are not required for p24rex production or virus replication and transformation.

Human T-cell leukemia virus types I (HTLV-I) and II (HTLV-II) have two nonstructural trans-acting regulatory genes, tax and rex, located in the 3' region of the viral genome. The tax gene product (HTLV-I p40tax and HTLV-II p37tax) is the transcriptional activator of the viral long terminal repeat. The rex gene encodes two protein products, p27rex/p21rex and p26rex/p24rex in HTLV-I and HTLV-II, respectively. Rex acts posttranscriptionally to facilitate accumulation of full-length gag/pol and singly spliced env mRNA in the cytoplasm of HTLV-infected cells. Previous studies showed that the first ATG of the rex gene is critical for Rex production and function. The importance of the internal ATGs to Rex function is not known. However, in vitro mutagenesis of the HTLV-I rex gene has provided indirect evidence which suggests that p21rex, and by analogy HTLV-II p24rex, results from initiation at an internal AUG of the tax/rex mRNA. By using an infectious molecular clone of HTLV-II, we investigated the importance of the internal ATGs of the rex gene on Rex protein production and function. Our results indicate that p24rex of HTLV-II is not initiated at an internal AUG and that the internal methionine codons are not crucial to the function of the rex gene and, ultimately, the transforming properties of the virus.

The Rex proteins of human T-cell leukemia virus type II differ by serine phosphorylation.

The Rex proteins of human T-cell leukemia virus types I and II (HTLV-I and HTLV-II) induce cytoplasmic expression of unspliced gag-pol mRNA and singly spliced env mRNA and are critical for virus replication. Two rex gene products, p27rex and p21rex of HTLV-I and p26rex and p24rex of HTLV-II, have been detected in HTLV-infected cells; however, the structural and biological relationship of the proteins has not been clearly elucidated. Endoproteinase digestion and phosphoamino acid analysis of HTLV-II Rex indicated that p24rex has the same amino acid backbone as p26rex and that the larger apparent molecular size of p26rex is attributable to serine phosphorylation.