[Survey of Helicobacter pylori levofloxacin and clarithromycin resistance rates and drug resistance genes in Ningxia, 2020-2022].

Objective: To explore the rate of Helicobacter pylori (Hp) resistance to levofloxacin and clarithromycin and the common mutation patterns of resistance genes in Ningxia, and to assess the concordance between phenotypic resistance and genotypic resistance. Methods: Cross-sectional study. Patients diagnosed with Hp infection in 14 hospitals in Ningxia region from February 2020 to May 2022 were retrospectively selected. Hp strains were isolated from gastric biopsy specimens of Hp-infected patients and subjected to phenotypic drug sensitivity testing and detection of resistance genes to analyze the rate of Hp resistance to levofloxacin and clarithromycin and the common mutation patterns of resistance genes in Ningxia region; and the concordance rate and Kappa concordance test were used to assess the concordance between phenotypic resistance and genotypic resistance. Results: A total of 1 942 Hp strains were isolated and cultured, and among the infections, 1 069 cases (55.0%) were male and 873 cases (45.0%) were female, aged (50.0±12.5) years (15-86 years). The rates of Hp resistance to levofloxacin and clarithromycin in Ningxia were 42.1% (818/1 942) and 40.1% (779/1 942), respectively, and the rate of dual resistance to both was 22.8% (443/1 942). The rate of resistance to levofloxacin and clarithromycin of Hp strains from female patients was higher than in male patients (levofloxacin: 50.4%(440/873) vs 35.4%(378/1 069); clarithromycin: 44.4%(388/873) vs 36.6%(391/1 069), both P<0.001). Among the GyrA gene mutations associated with levofloxacin resistance, the differences in mutation rate of amino acid at positions 87 and 91 were statistically significant in both drug-resistant and sensitive strains(both P<0.001), except for Asn87Thr. Hp strains were statistically significant for levofloxacin (Kappa=0.834, P<0.001) and clarithromycin (Kappa=0.829, P<0.001) had good concordance in resistance at the phenotypic and genotypic levels. Conclusion: The resistance of Hp to levofloxacin and clarithromycin in Ningxia region is severe, and there is good consistency between genotypic and phenotypic resistance.

[Whole transcriptome analysis and critical gene regulatory network analysis during Schistosoma japonicum infection and praziquantel treatment in mice].

OBJECTIVE: To investigate long non-coding RNA (lncRNA)-microRNA (miRNA)-messenger RNA (mRNA) interactions and identify the critical gene regulatory network during Schistosoma japonicum infections and praziquantel treatment using whole transcriptome sequencing. METHODS: A total of 110 male C57BL/6 mice were randomly divided into the control group, the infection group and the treatment group. Mice in the infection treatment and the control group were infected with S. japonicum cercariae via the abdomen, and liver specimens were sampled from 10 mice 3, 6, 8 weeks post-infection. Praziquantel treatment was given to mice in the treatment group 8 weeks post-infection, and liver specimens were sampled from 10 mice 2, 4, 6, 8, 10 weeks post-treatment. Total RNA was isolated from mouse liver specimens, and the transcriptome library was constructed for highthroughput whole transcriptome sequencing. The significant differentially expressed genes were subjected to functional annotations, Gene Ontology (GO) terms enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Correlation analysis of liver specimens was performed using R Corrplot and Himsc functions, and the lncRNAmiRNA-mRNA interaction network analysis was performed using R MixOmics and Himsc functions. RESULTS: There were 1 176 differentially expressed miRNAs, 5 270 differentially expressed mRNAs, and 2 682 differentially expressed lncRNAs between the infection group and the control group, 1 289 differentially expressed miRNAs, 7 differentially expressed mRNAs, and 69 differentially expressed lncRNAs between the treatment group and the infection group, and 1 210 differentially expressed miRNAs, 4 456 differentially expressed mRNAs, and 2 016 differentially expressed lncRNAs between the treatment group and the control group. Correlation analysis showed a higher correlation of gene expression between the treatment group and the control group. Principal component analysis showed obvious separate clustering between the infection group and the treatment group. The differentially expressed genes with significant relevance were significantly enriched in 24 GO terms, including arachidonic acid metabolic process, xenobiotic catabolic process, unsaturated fatty acid metabolic process, xenobiotic metabolic process, long-chain fatty acid metabolic process, and 8 KEGG metabolic pathways, including cholesterol metabolism, tyrosine metabolism, linoleic acid metabolism, retinol metabolism, and steroid hormone biometabolism. CONCLUSIONS: There were 23 mRNAs including Cyp2b9 and 14 lncRNAs including Rmrpr in the core position of the gene regulatory network, which may play a critical role in S. japonicum infections and praziquantel treatment, and 9 miRNAs including miR-8105 may serve as potential molecular markers for diagnosis of S. japonicum infections.

Transcriptome analysis of the bursa of Fabricius and thymus of laying ducks reveals immune gene expression changes underlying the impacts of stocking densities.

1. The thymus and bursa of Fabricius are important immune organs in poultry as they play essential roles in sustaining the normal immune function to maintain health. The following trial investigated whether the stocking density affected gene expressions in immune organs.2. Jinding ducklings were raised in either low or high density (4 or 8 birds/m2) conditions from four to 14 weeks of age, and were then slaughtered and tissues removed. Samples were subjected to high-throughput sequencing to sequence RNA extraction. After filtering calculations with R software, a total of 508 (thymus) and 1,356 (bursa of Fabricius) differentially expressed genes (DEGs) were identified, suggesting that stocking density has an effect on gene expression in duck immune organs.3. Out of a total of 112 immune factor genes and 112 immune pattern receptor genes in ducks, four thymus and 18 bursa of Fabricius genes were differentially expressed in ducks, which indicated that the change of stocking density could affect the expression of immune genes in poultry.

Baicalein inhibits the proliferative activity of human prostate cancer cell line PC3 by downregulating Ezrin.

We aimed to elucidate the inhibitory effect of Baicalein on proliferative ability in Prostate cancer (PCa) through downregulating Ezrin. Relative level of Ezrin in PCa tissues and adjacent ones was detected. After PC3 cells were induced with 20 or 40 µM Baicalein, changes in viability, cell cycle progression and apoptosis were assessed. Relative levels of CyclinD1, CDK4, P53 and P21 were examined by quantitative real-time polymerase chain reaction (qRT-PCR). Regulatory effects of Ezrin and Baicalein treatment on PC3 cells were evaluated. Finally, in vivo effects of Ezrin and Baicalein treatment on nude mice bearing PCa were detected. Ezrin was upregulated in PCa tissues relative to adjacent normal ones. Baicalein treatment decreased viability, arrested cell cycle and stimulated apoptosis in PC3 cells. Meanwhile, Baicalein treatment downregulated CyclinD1 and CDK4, while upregulating P53 and P21. Moreover, Ezrin was downregulated in Baicalein-treated PC3 cells. Knockdown of Ezrin synergistically stimulated the effects of Baicalein on cellular phenotypes of PC3 cells. In nude mice bearing PCa, Baicalein treatment decreased tumor volume and tumor weight, which were much more pronounced in those with in vivo knockdown of Ezrin. Baicalein treatment suppresses proliferative ability, arrests cell cycle and stimulates apoptosis in PCa cells through downregulating Ezrin.

Artificial modulation-free Pound-Drever-Hall method for laser frequency stabilization.

We have proposed an artificial modulation-free Pound-Drever-Hall (PDH) method for laser frequency stabilization and demonstrated it via two-color polarization spectroscopy of Rydberg electromagnetically induced transparency (EIT) resonance in a room-temperature rubidium vapor. Due to the unique error signal profile, the conventional PDH method owns a large capture range in laser frequency locking. Here we manually construct a PDH error signal via a linear combination of polarization spectroscopies of the Rydberg EIT resonances without and with a magnetic field applied. The artificial modulation-free PDH error signal owns a subnatural linewidth dispersion curve as well as a large capture range with which we successfully stabilize the laser to an absolute atomic frequency reference in a long running time, immune to environmental fluctuation and even manmade impulse perturbation. This method can provide an absolute frequency reference based on atomic transition while keeping similar locking ability to provide corrections for frequency fluctuations over a broad bandwidth as the conventional PDH.

[Application of digital 3D imaging technique in flap design before maxillofacial soft tissue reconstruction].

Objective:Using digital technique to design the preoperative flap of oral and maxillofacial soft tissue defect, to instruct the operation and to provide the basis for making the individual femoral flap. Method:We chosed 7 patients with oral and maxillofacial malignancies to undergo thin-line CT scans of lower extremities and analyzed the data of CTA examination to measure the diameter, shape and length of vascular pedicle. Then selected the appropriate perforation as the source of blood supply for flap. The location of perforation was marked on the skin to simulate flap removal range.During the surgery according to the preoperative mark, draw lines and mark flap size. After flap preparation is completed, suture the donor area directly and build in negative pressure drainage tube. Result:All reconstruction of the anterolateral femoral artery descending perforator flap can show the skin, muscle, blood vessels and their adjacent relationship, and according to the size of the defect area, using Mimics 17.0 software simulation to cut flap which should be the same as the defect area. The main perforating branches and trunks of seven skin flaps were consistent with pre-operative examination and all skin flaps survived; the wound surface and the donor were all in stage Ⅰ healing. After operation, patients were followed up for 1-6 months and the form, quality and function of skin flaps all recovered well. Conclusion:The location of lateral femoral circumflex artery descending limb and perforating branch were accurately located by digital technique three-dimensional reconstruction technology, showing the spatial relationship among skin, muscle, blood vessels and bones. By combining with Mimics 17.0 software, it can achieve three-dimensional and accurate design of anterolateral thigh flap, reduce the donor injury and improve the successful rate of skin flap.

Molecular cloning and characterization of a proliferating cell nuclear antigen gene by chemically induced male sterility in wheat (Triticum aestivum L.).

Although a number of studies have shown that chemical hybridizing agents (CHAs) affect anther growth and regulate cell-cycle progression, little is known about the molecular and cellular mechanisms involved. Proliferating cell nuclear antigen (PCNA) is an essential factor in DNA replication, and in many other processes in eukaryotic cells. In this study, the open reading frame of TaPCNA, the PCNA in wheat (Triticum aestivum L.), was cloned by reverse transcription polymerase chain reaction (RT-PCR). Sequence analysis revealed that this gene was 792-bp long and encoded a protein with 234 amino acids. Alignment of the TaPCNA-predicted sequence revealed a high degree of identity with PCNAs from other plant species. A subcellular localization assay indicated that TaPCNA was localized in the nucleus. The TaPCNA was cloned into the prokaryotic expression plasmid pET32a, and the recombinant plasmid was transformed into BL21 (DE3). TaPCNA expression was induced by 0.5 mM isopropyl-beta-D-thiogalactopyranoside and verified using sodium dodecyl sulfate polyacrylamide gel electrophoresis and western blot assays, which indicated that the fusion protein was successfully expressed. The gene involved in the G1-to-S transition, Histone H4, was downregulated by 1376- CIMS, which is a chemically induced male sterility line. However, a semi-quantitative RT-PCR revealed that TaPCNA expression was upregulated in 1376-CIMS. Our results suggest that CHAs (SQ-1) induce DNA damage in wheat anthers. DNA damage results in either the delay or arrest of cell-cycle progression, which affects anther development. This study will help to elucidate the mechanisms of SQ-1-induced male sterility.

Relationship between male sterility and ß-1,3-glucanase activity and callose deposition-related gene expression in wheat (Triticum aestivum L.).

In previous studies, we first isolated one different protein ß-1,3-glucanase using two-dimensional electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry from normal wheat (Triticum aestivum L.) and chemical hybridization agent-induced male sterility (CIMS) wheat. In this experiment, ß-1,3-glucanase activity and the expression of a callose deposition-related gene, UDP-glucose phosphorylase (UGPase), were determinate in normal, CIMS, and genetic male sterility (GS) wheat. ß-1,3-glucanase activity was significantly different between the fertile and sterile lines during callose synthesis and degradation, but there was no difference between CIMS and GS wheat. The UGPase gene of callose deposition was highly expressed in the meiophase and sharply decreased in the tetrad stage. However, the expression of the UGPase gene was significantly different between the fertile and sterile lines. These data indicated that ß-1,3-glucanase activity and the expression of the UGPase gene play important roles in the male sterility of wheat. Consequently, pollen mother cells (PMCs) might degenerate at the early meiosis stage, and differences in UGPase gene expression and ß-1,3-glucanase activity might eventually result in complete pollen collapse. In addition, the critical period of anther abortion might be the meiosis stage to the tetrad stage rather than what we previously thought, the mononuclear period.

Homocysteine induces blood vessel global hypomethylation mediated by LOX-1.

Homocysteine (Hcy) is an independent risk factor of atherosclerosis through its involvement with the methionine cycle. In this study, we aimed to determine the blood vessel global methylation rate in Hcy-induced atherosclerosis in apolipoprotein-E-deficient (ApoE-/-) mice, and to explore the possible mechanism of this change in endothelial cells. ApoE-/- mice were divided into a hyperlipidemia (HLP) group, a hyperhomocysteinemia (HHcy) group, and an HHcy + folate + vitamin B12 (HHcy+FA+VB) group. Wild-type C57BL/6J mice were prepared as controls. Total Hcy, lipids, S-adenosylmethionine (SAM), and S-adenosylhomocysteine (SAH) contents in serum were measured with an automatic biochemistry analyzer and high-performance liquid chromatography. Methylation of B1 repetitive elements in blood vessels was tested using nested methylation-specific-polymerase chain reaction (nMS-PCR). Endothelial cells (ECs) were pretreated with Hcy or by adding FA and VB. Lectin-like oxidized LDL receptor-1 (LOX-1) expressions were determined by quantitative PCR, Western blot, and nMS-PCR. The HHcy group displayed severe HLP and HHcy. SAM and SAH contents were also elevated in the HHcy group compared with other groups. Methylation of B1 repetitive elements was significantly increased in the HHcy group (0.5050 ± 0.0182) compared to the HLP (0.5158 ± 0.0163) and control (0.5589 ± 0.0236) groups. mRNA and protein expressions of LOX-1 increased (0.2877 ± 0.0341, 0.6090 ± 0.0547), whereas methylation expression decreased (0.5527 ± 0.0148) after 100 µM Hcy stimulation in ECs. In conclusion, Hcy-induced atherosclerosis was closely associated with induced hypomethylation status in the blood vessel, and this process was partially mediated by LOX-1 DNA methylation.

Tranexamic acid is associated with less blood transfusion in off-pump coronary artery bypass graft surgery: a systematic review and meta-analysis.

OBJECTIVE: Tranexamic acid reduces blood loss and transfusion in on-pump coronary artery bypass graft (CABG) surgery. Compared with on-pump, off-pump surgery is associated with less blood loss and transfusion. Therefore, tranexamic acid may be less effective for off-pump surgery, and its safety profile may be different in this setting. The aim of this study was to determine the efficacy and safety of tranexamic acid for off-pump CABG surgery. DESIGN: Systematic review and meta-analysis. SETTING: University of Edinburgh. INTERVENTIONS: The administration of tranexamic acid. METHODS: A systematic review of randomized controlled trials administering tranexamic acid to patients undergoing off-pump CABG surgery. A meta-analysis of 24-hour blood loss, postoperative allogeneic transfusion, and thromboembolic events. MEASUREMENTS AND MAIN RESULTS: Eight trials were identified. The lack of appropriate data limited the meta-analysis on blood loss. Tranexamic acid significantly reduced the overall risk of allogeneic blood component transfusion (risk ratio = 0.47; 95% confidence intervals, 0.33-0.66; p < 0.0001) and packed red blood cell transfusions (risk ratio = 0.51; 95% CI, 0.36-0.71; p = 0.0001). No association was found between tranexamic acid and myocardial infarction, stroke, or pulmonary embolism. Population sizes of meta-analyses ranged from 466 to 544. CONCLUSIONS: Tranexamic acid reduces blood transfusion after off-pump surgery. Although no association with adverse events was found, the population sample size was too small to detect rare but clinically significant adverse events. A well-designed randomized controlled trial with an appropriate sample size is required to confirm tranexamic acid effectiveness and safety in off-pump CABG surgery.

Clinical trial simulation: a review.

Modeling and simulation in general, and specifically clinical trial simulation (CTS), have been recognized by the (larger) pharmaceutical companies and regulatory authorities as being pivotal to improving the efficiency of the drug development process. This includes the use of CTS to learn about drug effectiveness and safety and to optimize trial designs at the various stages of development. By reviewing papers published during the period January 2000-January 2010, this paper discusses recent developments with respect to methodology, applications, and lessons learned in the use of CTS in the development and clinical use of specific drugs. It is expected that future CTS experiments will be aided by the hybridization of optimal design methods with computationally intensive stochastic simulations. This will take advantage of optimizing the experimental design and leave the task of evaluating the probable real-world performance of a limited number of candidate trial designs and analysis procedures.

A new glucoside from Selaginella sinensis.

A new glucoside, selaginoside (1), together with two known compounds, hinokiflavone (2) and 2,3-dihydroamentoflavone (3), were isolated from the whole plants of Selaginella sinensis. Their structures were elucidated by means of spectroscopic methods.

In vitro evaluation of secoiridoid glucosides from the fruits of Ligustrum lucidum as antiviral agents.

Six secoiridoid glucosides, lucidumoside C (1), oleoside dimethylester (2), neonuezhenide (3), oleuropein (4), ligustroside (5) and lucidumoside A (6), isolated from the fruits of Ligustrum lucidum (Oleaceae), were examined in vitro for their activities against four strains of pathogenic viruses, namely herpes simplex type I virus (HSV-1), influenza type A virus (Flu A), respiratory syncytial virus (RSV) and parainfluenza type 3 virus (Para 3). Antiviral activities were evaluated by the cytopathic effect (CPE) inhibitory assay. The purpose was to check if the antioxidative potency of these glucosides correlated with their antiviral potency. Results showed that none of the glucosides had any significant activity against HSV-1 and Flu A. Oleuropein, however, showed significant antiviral activities against RSV and Para 3 with IC50 value of 23.4 and 11.7 microg/ml, respectively. Lucidumoside C, oleoside dimethylester and ligustroside showed potent or moderate antiviral activities against Para 3 with IC50 values of 15.6-20.8 microg/ml. These results also documented that the anti-oxidative potency of these secoiriodoid glucosides was not directly related to their antiviral effects.

New antiviral cassane furanoditerpenes from Caesalpinia minax.

A bioassay-guided study led to the isolation of five new cassane furanoditerpenes, designated as caesalmin C (1), D (2), E (3), F (4), and G (5), along with stigmasterol (6) from the seeds of Caesalpinia minax. The (1)H and (13)C NMR spectra were completely assigned by using a combination of 2D NMR analyses. The structures of all five furanoditerpenes were confirmed by X-ray analyses. The structure of 6 was verified by X-ray analysis for the first time. The bioassay results showed that the anti-Para3 virus activity of tetracyclic furanoditerpenoids 1-4 is more potent than that of the furanoditerpenoid lactone 5, which is in turn better than 6. As the major components of the plant possess significant potent activity, it may be feasible to develop new antiviral agents from this source.

A novel 1:1 complex of potassium mikanin-3-O-sulfate with methanol.

Mikanin-3-O-sulfate (1), in the form of its potassium salt, together with mikanin (2) and alpinetin (3) were isolated from Mikania micrantha. The crystal structures of K(1) x CH3OH, 2 and 3 x H2O were established by X-ray crystallography. The potassium ions in K(1) x CHO3H are bridged by O5, O7 and O8 to form a chain of face-sharing KO8 coordination polyhedra, from which the aglycon units are outstretched to form a polymeric molecular column. Adjacent molecular columns are linked by pi-pi stacking between parallel, intercalating aglycon units to form layers matching the (101) family of planes, which are further interconnected into a three-dimensional supramolecular assembly. Sulfation at 3-OH induced better co-planarity and conjugation of the rings.

Furanoditerpenoid lactones from the seeds of Caesalpinia minax Hance.

Two furanoditerpenoid lactones, were isolated from the seeds of Caesalpinia minax Hance. Their structures and stereochemistry have been established by spectral data and single crystal X-ray analysis.

Antiviral amentoflavone from Selaginella sinensis.

Amentoflavone and three other flavonoids were isolated from the ethanol extract of Selaginella sinensis. Amentoflavone showed potent antiviral activity against respiratory syncytial virus (RSV), with an IC50 of 5.5 microg/ml. The contents of amentoflavone in nine species of Selaginella were determined by reversed-phase HPLC. S. sinensis showed a higher content of 1.13%.

[The development of a original multifunctional peritoneal biopsy needle].

A multifunctional peritoneal biopsy needle is introduced in this paper, which can be used for peritoneal biopsy, brush biopsy and routine, biochemical and cytological examination. The clinical applications show that it could raise markedly the diagnosis rate of ascites, and especially it has important clinic diagnosis value for tuberculous and cancer ascites, and primary peritonitis complicated by hepatocirrhosis. It is simple, safe and practical to operate this needle.

Performance of a sulfide-oxidizing expanded-bed reactor supplied with dissolved oxygen.

The performance of a new sulfide-oxidizing, expanded-bed bioreactor is described. To stimulate the formation of well-settleable sulfur sludge, which comprises active sulfide-oxidizing bacterial biomass and elemental sulfur, the aeration of the liquid phase and the oxidation of sulfide to elemental sulfur are spatially separated. The liquid phase is aerated in a vessel and subsequently recirculated to the sulfide-oxidizing bioreactor. In this manner, turbulencies due to aeration of the liquid phase in the bioreactor are avoided. It appeared that, under autotrophic conditions, almost all biomass present in the reactor will be immobilized within the sulfur sludge which consists mainly of elemental sulfur (92%) and biomass (2.5%). The particles formed have a diameter of up to 3 mm and can easily be grinded down. Within time, the sulfur sludge obtained excellent settling properties; e.g., after 50 days of operation, 90% of the sludge settles down at a velocity above 25 m h(-1) while 10% of the sludge had a sedimentation velocity higher than 108 m h(-1). Because the biomass is retained in the reactor, higher sulfide loading rates may be applied than to a conventional "free-cell" suspension. The maximum sulfide-loading rate reached was 14 g HS(-) L(-1) d(-1), whereas for a free-cell suspension a maximum loading rate of 6 g HS(-) L(-1) d(-1) was found. At higher loading rates, the upward velocities of the aerated suspension became too high so that sulfur sludge accumulated in the settling zone on top of the reactor. When the influent was supplemented with volatile fatty acids, heterotrophic sulfur and sulfate reducing bacteria, and possibly also (facultatively) heterotrophic Thiobacilli, accumulated within the sludge. This led to a serious deterioration of the system; i.e., the sulfur formed was increasingly reduced to sulfide, and also the formation rate of sulfur sludge declined. (c) 1997 John Wiley & Sons, Inc.