[Exploration on mechanisms of Xiaoyao Powder in treating atherosclerosis and depressive disorder with concept of "treating different diseases with same method" based on network pharmacology].

The aim of this paper was to explore the mechanism of Xiaoyao Powder in treating atherosclerosis and depressive disorder with concept of "treating different diseases with same method" based on network pharmacology. TCMSP(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform) and SymMap databases were used to search all the chemical components and targets related to Xiaoyao Powder. After preliminary screening, the network of "herbs-compounds-targets" was constructed. Through DisGeNET, CTD(Comparative Toxicogenomics Database) and TTD(Therapeutic Target Database), the targets of atherosclerosis and depressive disorder were obtained. The common targets were obtained by intersecting the herbal targets and disease targets. In order to screen the key common targets, STRING and Cytoscape were used to analyze the protein-protein interaction of common targets. BioGPS was used to obtain their distribution information in organs and tissues. Gene ontology(GO) and Kyoto encyclopedia of genes and genomes(KEGG) analysis were conducted through Metascape. About 1 355 compounds of Xiaoyao Powder were found by TCMSP and Symmap database; 161 active compounds were screened out according to standard of oral bioavailability≥30% and drug like index≥0.18; 274 herbal targets were obtained and the "herbs-compounds-targets" network was constructed. About 1 004 atherosclerosis targets and 578 depressive disorder targets were obtained, and 37 common targets were obtained after intersection with herbal targets. By using STRING and Cytoscape for protein-protein interaction analysis, 18 key targets were screened. BioGPS showed that the key common targets were mainly distributed in heart, amygdala, pineal, liver and smooth muscle. Metascape was used for GO enrichment analysis and the results showed that there were 929 biological processes, 25 cell components and 23 molecular functions. Enrichment ana-lysis of KEGG showed that there were 108 signal pathways such as AGE-RAGE, HIF-1, FoxO, Th17 cell differentiation and IL-17 signal pathways, which were mainly related to neuroendocrine system, metabolism, immune inflammation and oxidative stress. In conclusion, the main mechanism of Xiaoyao Powder in treating atherosclerosis and depressive disorder with concept of "treating different diseases with same method" was related to neuroendocrine system, metabolism, immune inflammation and oxidative stress-related signal pathway, providing reference for further experimental verification, potential pharmacological mechanism and clinical application.

[Study on syndrome differentiation of ancient arthralgia syndrome and drug use rules based on latent structure model].

In this study, the ancient doctors' concentrated prescriptions for arthralgia syndrome were collected and Lantern 3.2 software was used to construct and analyze the latent structure model of ancient arthralgia syndrome. A total of 10 syndrome types of ancient arthralgia syndrome were obtained, namely, cold-coagulation meridian syndrome(partial Yang deficiency), cold-dampness obstruction syndrome(cold weight), damp-heat obstruction syndrome, liver and kidney deficiency syndrome, essence deficiency syndrome, phlegm and blood stasis syndrome, spleen and stomach Qi deficiency syndrome, dampness evil accumulation syndrome, arthralgia meridian syndrome, cold-dampness binding lung syndrome, meridian obstruction syndrome, and wind-cold-dampness arthralgia syndrome(both wind-cold and dampness). This syndrome differentiation method further strengthened the thought on eight-class syndrome differentiation, and showed the characteristics of different syndrome differentiation types of the core drugs in the latent structure model, which was mainly reflected in paying attention to removing blood stasis and eliminating phlegm with use of pungent taste products having dispersing effect, emphasizing the effect of nourishing Yin and generating marrow to supplement congenital deficiency, invigorating spleen and replenishing Qi to regulate the spleen and stomach functions, nourishing vital energy and expelling evil. In addition, Python 3.7 software was used to mine the frequent item sets and association rules for the frequently used drugs for ancient arthralgia syndrome. The drug compatibility law is characterized by making good use of blood-nourishing drugs. Blood-activating drugs are often compatible with cold-dispelling drugs, liver and kidney tonifying drugs, and wind-dampness-dispelling drugs, following the compatibility principle of dispelling wind and dispelling cold as well as tonifying liver and kidney. The induction of syndrome differentiation types of ancient arthralgia syndrome and the discussion on the characteristics of drug use and compatibility law provide a new way for clinical syndrome differentiation and treatment, with certain reference value for the research and development of new Chinese patent medicines.

Simulated microgravity reduces proliferation and reorganizes the cytoskeleton of human umbilical cord mesenchymal stem cells.

The cytoskeleton plays a key role in cellular proliferation, cell-shape maintenance and internal cellular organization. Cells are highly sensitive to changes in microgravity, which can induce alterations in the distribution of the cytoskeletal and cell proliferation. This study aimed to assess the effects of simulated microgravity (SMG) on the proliferation and expression of major cell cycle-related regulators and cytoskeletal proteins in human umbilical cord mesenchymal stem cells (hucMSCs). A WST-1 assay showed that the proliferation of SMG-exposed hucMSCs was lower than a control group. Furthermore, flow cytometry analysis demonstrated that the percentage of SMG-exposed hucMSCs in the G0/G1 phase was higher than the control group. A western blot analysis revealed there was a downregulation of cyclin A1 and A2 expression in SMG-exposed hucMSCs as well. The expression of cyclin-dependent kinase 4 (cdk4) and 6 (cdk6) were also observed to be reduced in the SMG-exposed hucMSCs. The total nuclear intensity of SMG-exposed hucMSCs was also lower than the control group. However, there were no differences in the nuclear area or nuclear-shape value of hucMSCs from the SMG and control groups. A western blot and quantitative RT-PCR analysis showed that SMG-exposed hucMSCs experienced a downregulation of bata-actin and alpha-tubulin compared to the control group. SMG generated the reorganization of microtubules and microfilaments in hucMSCs. Our study supports the idea that the downregulation of major cell cycle-related proteins and cytoskeletal proteins results in the remodeling of the cytoskeleton and the proliferation of hucMSCs.

CT imaging of inflatable penile prosthesis complications: a pictorial essay.

PURPOSE: Inflatable penile prostheses (IPPs) are widely used in the United States, for patients with erectile dysfunction refractory to other treatments. Complications subsequent to IPP insertion include infection, hematoma, fluid leak, component (cylinder/pump/reservoir) complications, and retained previous IPP components. Radiologists are often called upon to identify and characterize these complications utilizing CT prior to intervention. Our paper aims to provide a guide to familiarize radiologists with normal IPP component imaging and review the CT findings of commonly encountered complications. METHODS: In this study, we retrospectively reviewed CT reports with descriptions of IPPs from 108 patients. We collected CT images of normal IPP components as well as reevaluated the CT findings of 33 patients with an IPP complication and correlated with immediately subsequent operative report. RESULTS: The CT appearance of appropriately positioned normal IPP components in asymptomatic patients and each complication were described and compared to previous literature. CONCLUSIONS: CT is a very useful modality to assess an IPP-related complication. It is inexpensive, fast, and immediately available in emergent situations, e.g., infection, hematoma, and component erosion. Additionally, CT is very sensitive and makes it easier to diagnose a system leak. It can identify most cylinder complications and pump malposition and can be extremely helpful to the surgeon in preoperative planning if revision is needed. After this review, the radiologist should be able to identify normal IPP components and their complications.

MiR-130b functions as a tumor promoter in glioma via regulation of ERK/MAPK pathway.

OBJECTIVE: To investigate the miR-130b expression in patients with glioma and to analyze its role and underlying molecular mechanism on the carcinogenesis. PATIENTS AND METHODS: The expression levels of miR-130b were detected with quantitative Real-time PCR. The relationship between miR-130b expression and clinicopathologic characteristics were analyzed. MiR-130b inhibitor was transfected into glioma cell lines to investigate its role in HCC. MTT assays were conducted to explore the impact of miR-130b down-expression on the proliferation of human glioma cells. Cell cycle and cell apoptosis assays were performed using flow cytometry. Levels of ERK/MAPK pathway related proteins were evaluated by Western blotting. Data were analyzed using the 2-ΔΔCT method through student's t-test via the GraphPad Prism software (La Jolla, CA, USA). RESULTS: The expression of miR-130b was markedly upregulated in glioma cell lines and tissues, and high miR-130b expression was significantly associated with advanced WHO grade (p = 0.022) and low Karnofsky performance score (p = 0.001). In addition, downregulation of miR-130b inhibited the proliferation of glioma cells and induced cell-cycle arrest and cells apoptosis in vivo. Importantly, ERK/MAPK pathway was found to be inactivated in the glioma cell lines after miR-130b knockout experiment. CONCLUSIONS: The current data indicated that miR-130b may play a critical role in the progression of glioma via ERK/MAPK signaling cascades, suggesting that it may be a useful therapeutic agent in glioma patients.

Biomedical journals lack a consistent method to detect outcome reporting bias: a cross-sectional analysis.

WHAT IS KNOWN AND OBJECTIVE: An increasing amount of recently published literature has implicated outcome reporting bias (ORB) as a major contributor to skewing data in both randomized controlled trials and systematic reviews; however, little is known about the current methods in place to detect ORB. This study aims to gain insight into the detection and management of ORB by biomedical journals. METHODS: This was a cross-sectional analysis involving standardized questions via email or telephone with the top 30 biomedical journals (2012) ranked by impact factor. The Cochrane Database of Systematic Reviews was excluded leaving 29 journals in the sample. RESULTS: Of 29 journals, 24 (83%) responded to our initial inquiry of which 14 (58%) answered our questions and 10 (42%) declined participation. Five (36%) of the responding journals indicated they had a specific method to detect ORB, whereas 9 (64%) did not have a specific method in place. The prevalence of ORB in the review process seemed to differ with 4 (29%) journals indicating ORB was found commonly, whereas 7 (50%) indicated ORB was uncommon or never detected by their journal previously. The majority (n = 10/14, 72%) of journals were unwilling to report or make discrepancies found in manuscripts available to the public. Although the minority, there were some journals (n = 4/14, 29%) which described thorough methods to detect ORB. WHAT IS NEW AND CONCLUSION: Many journals seemed to lack a method with which to detect ORB and its estimated prevalence was much lower than that reported in literature suggesting inadequate detection. There exists a potential for overestimation of treatment effects of interventions and unclear risks. Fortunately, there are journals within this sample which appear to utilize comprehensive methods for detection of ORB, but overall, the data suggest improvements at the biomedical journal level for detecting and minimizing the effect of this bias are needed.

Disruption of ClC-2 expression is associated with progressive neurodegeneration in aging mice.

Heterozygous mutations in ClC-2 have been associated in rare cases with increased susceptibility to generalized, idiopathic epilepsy. Initially, it was hypothesized that mutations in ClC-2 may be associated with epilepsy due to a direct role for ClC-2 in the modification of hippocampal neuronal excitability. However, the absence of an overt seizure-susceptibility phenotype in young ClC-2 knockout (KO) mice rendered this hypothesis- implausible. A recent study of older ClC-2 KO mice (>6 months) revealed abnormalities in the myelin of central axons and a subtle defect in the neuronal function in the central auditory pathway. These findings prompted us to re-examine hippocampal neuron morphology and excitability in older ClC-2 KO mice. Interestingly, electrocorticographic recordings obtained in older mice revealed spontaneous interictal spikes which are a marker of perturbed hippocampal neurotransmission with a resultant increase in excitation. This electrophysiological defect was associated with astrocyte activation and evidence of neuronal degeneration in the CA3 region of the hippocampus of these older mice. Together, these findings raise the possibility that ClC-2 expression plays a subtle neuroprotective role in the aging hippocampus.

Antifouling coatings for optoelectronic tweezers.

Optoelectronic tweezers enables parallel manipulation of individual single cells using optical addressing and optically induced dielectrophoretic force. This provides a useful platform for performing a variety of biological functions, such as cell manipulation, cell sorting, and cell electroporation. However, in order to obtain more reliable cellular manipulation, especially of adherent mammalian cells, antifouling coatings need to be used to avoid non-specific cell adherence. Two antifouling coatings are discussed here, which can reduce the amount of non-specific adherence by as much as a factor of 30.

Developmental neurotoxicity role of cyclophosphamide on post-neural tube closure of rodents in vitro and in vivo.

To evaluate the possible developmental role of cyclophosphamide on post-neural tube closure of rodents, the 46 pregnant rats of 2.5-3 months old were randomly divided into five groups. On the 13th day of gestation, the rats in control and experimental groups were treated with physiological saline and cyclophosphamide (7.5, 10.5, 12.5 and 15.0 mg/kg bw), respectively. On the 14th day of gestation, three rats each selected randomly from control and experimental (12.5 mg/kg bw) group, respectively, were executed and the embryo brains were checked using both light microscope and transmission electron microscope. Neurocytotoxic effects of cyclophosphamide were determined using tetrazolium dye (MTT) assay, single cell gel electrophoresis, flow cytometry and scanning electron microscope. Compared with control group, the malformation incidence of the experimental groups (except the group 7.5 mg/kg bw) was significantly higher, especially in 12.5 mg/kg bw group (malformation incidence 98%, lethality 0). Neural tube serial section of fetus showed the apoptotic morphological features after 24 h cyclophosphamide administrated. Cyclophosphamide (8 microg/ml) decreased the growth and viability of neurons, damaged nuclear DNA and induced early apoptotic morphological changes. Our study shows that the teratogenesis of cyclophosphamide after neural tube closure are involved in cyclophosphamide-induced neuronal apoptosis and DNA damage.

Subcellular localization, expression patterns, SNPs and association analyses of the porcine HUMMLC2B gene.

Myosin regulatory light chain (MLC) regulates myofilament activation via phosphorylation by Ca(2+) dependant myosin light chain kinase. In order to further understand the functions of the porcine fast myosin regulatory light chain gene (HUMMLC2B) in muscle, the subcellular localization, the temporal and spatial distributions of its gene product were analyzed, and the association between the presence of specific polymorphisms and commercial meat traits in pig was also examined. HUMMLC2B was demonstrated to localize both in the cytoplasm and the nucleus by confocal fluorescence microscopy. Real-time PCR further revealed HUMMLC2B expression variation in a waveform manner in the skeletal muscle of both Chinese Tongcheng and Western Landrace pig breeds at days 33, 65 and 90 post coitum (pc). After birth, the expression levels of HUMMLC2B were also found to decrease gradually with age. Our spatial expression analysis showed that HUMMLC2B was highly expressed in the semitendinosus, gastrocnemius, biceps femoris and longissimus dorsi muscles. In contrast, only low levels of expression of this gene were evident in fat, and no expression was detectable in brain, heart, kidney, lung, liver, lymph node, spleen, stomach, or in either large or small intestine. A total of 23 potential polymorphisms, comprising 3 exonic and 20 intronic, were detectable in the porcine HUMMLC2B gene and the G1094A, T1513C, G1876A and T2005G polymorphisms were further analyzed. The significant associations between the T1513C, G1876A and T2005G polymorphisms with marbling score, dressing percent and meat color, respectively, were identified (P < 0.05). Associations with the percentage of leaf fat could also be demonstrated by analysis of haplotypes harboring these three polymorphisms. Our current results thus shed further light on the roles and functions of the HUMMLC2B gene in muscle.

Sodium selenite induces apoptosis in cultured cortical neurons with special concomitant changes in expression of the apoptosis-related genes.

Sodium selenite was used to examine whether selenium compound is able to trigger apoptotic degeneration in cultured cortical neurons in vitro and to explore the detailed changes in expression of the related genes during the apoptotic processes using molecular biological and flow cytometric examinations. The results indicated that: (1) cortical neurons treated with sodium selenite with different dosages (0.0008, 0.004, 0.0200, 0.1000, and 0.5000 microM) and different exposure times (2, 4, 24, and 48 h) exhibited dose- and time-dependent apoptotic processes as revealed by typical DNA ladder formation detected by agarose gel electrophoresis; (2) the internucleosomal DNA fragmentation detected by flow cytometric examination showed a prominent peak of hypodiploid DNA contents as early as 4h after exposure of 0.1 microM sodium selenite; (3) the DNA fragmentation induced by sodium selenite as revealed by the above two examinations could be blocked by aurintricarboxylic acid; (4) the transcriptions of mRNAs related to bcl-2, bax, c-fos, p53, and acetylcholinesterase (AChE) genes, as detected by RT-PCR assays, showed down-regulation for bcl-2 and up-regulation for bax, c-fos, p53, and AChE genes after exposure of sodium selenite. This study suggests that the sodium selenite is effective for inducing apoptosis in cultured cortical neurons and that relevant changes in expression of several apoptosis-related genes might further our understanding of the mechanism(s) that initiates and maintains the apoptotic processes.

Site-directed mutagenesis of the hinge region of nisinZ and properties of nisinZ mutants.

To study the role of the hinge region in nisin and to obtain mutants that exhibit altered or new biological activities and functional properties, we changed certain amino acids in the hinge region by performing site-directed mutagenesis with the nisinZ structural gene ( nisZ). The results showed that the nisinZ mutants had decreased antimicrobial activities against Micrococcus flavus NCIB8166 and Streptococcus thermophilus. Interestingly, compared with wild nisinZ, mutant N20K nisinZ and M21K nisinZ displayed antimicrobial activity against gram-negative Shigella, Pseudomonas and Salmonella; and they had a higher solubility than wild-type nisinZ. At pH 8, the solubilities of N20K nisinZ and M21K nisinZ were, respectively, three-fold higher and five-fold higher than that of nisinZ. Mutant N20Q nisinZ and M21G nisinZ were considerably more stable than nisinZ at higher temperatures and neutral or alkaline pH. These mutants provided information that the central hinge region in nisinZ plays an important role in providing the conformational flexibility required for the antimicrobial activity on the membrane. Our finding documented that it may well be worth considering the construction of the new nisin mutants with changed inhibitory activity against a wide range of gram-negative bacteria and the improvement of functional properties by site-directed mutagenesis.

[cDNA cloning and sequence analysis of human lactoferrin].

Human lactoferrin (hLF) cDNA was amplified by RT-PCR from normal human mammary tissue obtained from Daxing County of Beijing City of China, and then subcloned into pGEM-T vector. hLF cDNA sequence was determined, which consists of 2136 bp. Comparison with five other hLF cDNA sequences registered in GenBank shows 99% homology in DNA sequence. However, there are two base substitutions (nucleotide 1740 G-->C, nucleotide 1756 T-->C), one of which subsequently leads to an amino acid change (residue 580 Glu-->Asp).

A monomer is the minimum functional unit required for channel and ATPase activity of the cystic fibrosis transmembrane conductance regulator.

The cystic fibrosis transmembrane conductance regulator (CFTR) normally functions as a phosphorylation-regulated chloride channel on the apical surface of epithelial cells, and lack of this function is the primary cause for the fatal disease cystic fibrosis (CF). Previous studies showed that purified, reconstituted CFTR can function as a chloride channel and, further, that its intrinsic ATPase activity is required to regulate opening and closing of the channel gate. However, these previous studies did not identify the quaternary structure required to mediate conduction and catalysis. Our present studies show that CFTR molecules may self-associate in CHO and Sf9 membranes, as complexes close to the predicted size of CFTR dimers can be captured by chemical cross-linking reagents and detected using nondissociative PAGE. However, CFTR function does not require a multimeric complex for function as we determined that purified, reconstituted CFTR monomers are sufficient to mediate regulated chloride conduction and ATPase activity.

[Fusion expression of a peptide antibiotic-apidaecin gene in Lactococcus lactis].

The ubiquitin fusion of apidaecin was expressed in Lactococcus lactis, using a novel nisin-inducible expression system. After induction, a specific band could be detected in the extracts of the host strain by Tricine-SDS-PAGE and Western blotting. Production of the fusion was up to 7.2% of the total soluble protein of the host strain. While the fusion was cut by ubiquitin specific protease-UBP1, the product had distinct antibacterial activity.

[Cloning and expression of nisZ gene in Lactococcus lactis].

The gene encoding the precursor of nisin was amplified by PCR using the lambda HJ-3 DNA as the template, which contained the entire nisin biosynthesis gene cluster from Lactococcus lactis AL2 with high yield of nisin, and was cloned into pMG36e. The recombinant plasmid pHJ201 was introduced into Lactococcus lactis NZ9800 by electroporation. pHJ201 is very stable in L. lactis NZ9800. Antimicrobial activity test and Tricine-SDS-PAGE analysis revealed that L. lactis NZ9800 harbouring pHJ201 restored ability of nisin production, but the production level was markedly lower than L. lactis AL2. The result of DNA sequence analysis indicated that Nisin Z is produced by L. lactis AL2.

Perturbation of the pore of the cystic fibrosis transmembrane conductance regulator (CFTR) inhibits its atpase activity.

Mutations in the cystic fibrosis gene coding for the cystic fibrosis transmembrane conductance regulator (CFTR) lead to altered chloride (Cl(-)) flux in affected epithelial tissues. CFTR is a Cl(-) channel that is regulated by phosphorylation, nucleotide binding, and hydrolysis. However, the molecular basis for the functional regulation of wild type and mutant CFTR remains poorly understood. CFTR possesses two nucleotide binding domains, a phosphorylation-dependent regulatory domain, and two transmembrane domains that comprise the pore through which Cl(-) permeates. Mutations of residues lining the channel pore (e.g. R347D) are typically thought to cause disease by altering the interaction of Cl(-) with the pore. However, in the present study we show that the R347D mutation and diphenylamine-2-carboxylate (an open pore inhibitor) also inhibit CFTR ATPase activity, revealing a novel mechanism for cross-talk from the pore to the catalytic domains. In both cases, the reduction in ATPase correlates with a decrease in nucleotide turnover rather than affinity. Finally, we demonstrate that glutathione (GSH) inhibits CFTR ATPase and that this inhibition is altered in the CFTR-R347D variant. These findings suggest that cross-talk between the pore and nucleotide binding domains of CFTR may be important in the in vivo regulation of CFTR in health and disease.

[The construction and application of promoter-signal peptide probe vector of lactic acid bacteria].

A promoter-signal peptide probe vector of lactic acid bacteria, designated pZLB based on pMG36e has been constructed. The vector contained a reporterd gene encoding for beta-lactamase, which lacked a promoter and a functional signal sequence. Sau3A restriction fragments (80-400 bp) from L. lactis ATCC 7962 total DNA were cloned into BamHI site of pZLB. A lot of different size framents were selected. These fragments promoted transcription and secretion of beta-lactamase in L. lactis. Sequence analysis revealed that inserted fragment in pZLBe7 contained typical promoter, SD sequence and atypical signal sequence.

[Isolation and characterization of a plasmid pTS50, which encodes nisin resistance determinant in Lactococcus lactis TS1640].

Three nisin-resistant Lactococcus lactis strains were screened from 197 samples of fresh milk on a selective medium (M17) supplemented with nisin, lactose and bromocresol purple, and were confirmed to have the nisin resistance determinant (nsr) by PCR amplification. Physiological and biochemical tests as well as Lactococcus lactis specific 16S rDNA sequence analysis revealed that the three strains all belong to Lactococcus lactis subsp. lactis. A large plasmid, pTS50, was identified in L. lactis subsp. lactis TS1640 which encodes resistance to nisin following electroporation of the total plasmids DNA into L. lactis MG1363. The molecular weight of plasmid pTS50 was estimated to be 47 kb by restriction analysis of BamHI, EcoRI, HindIII, NcoI, PstI and nsr was localized on a 1.9 kb EcoRI fragment by Southern hybridization.

[Isolation and identification of the broad spectrum antimicrobial peptide AP311 produced by Lactobacillus acidophilus].

The heat stable antimicrobial peptide (AP311) produced by Lactobacillus acidophilus was isolated and identified. The AP311 has broad spectrum of inhibition including many Gram-positive and Gram-negative bacteria. The inhibition activity of AP311 was lost upon treatment with trypsin, subtilisin, proteanase K, chymotrypsin and pepsin. Inhibition activity of AP311 was decreased with pH increasing. The AP311 is very stable at acidic condition (pH2-4) even if heating at 100 degrees C for 30 min. But it was inactivated at basic pH (pH12) and the activity was restored completely upon reversion to acidic. The AP311 was not precipitated and inactivated by various organic solvents, except n-butanol. When the precipitate was redissolved in 0.02 mol/L HC1, its activity was restored. Based on its proteinaceous nature, broad spectrum of inhibition, we propose that AP311 should be considered the broad spectrum antimicrobial peptide.