Characteristics of phenotypic antibiotic resistance of Helicobacter pylori and its correlation with genotypic antibiotic resistance: A retrospective study in Ningxia.

BACKGROUND: Geographic differences exist in the antibiotic resistance patterns of Helicobacter pylori. Personalized treatment regimens based on local or individual resistance data are essential. We evaluated the current status of H. pylori resistance in Ningxia, analyzed resistance-related factors, and assessed the concordance of phenotypic and genotypic resistance. METHODS: Strains were isolated from the gastric mucosa of patients infected with H. pylori in Ningxia and relevant clinical information was collected. Phenotypic antibiotic susceptibility assays (Kirby-Bauer disk diffusion) and antibiotic resistance gene detection (Sanger sequencing) were performed. RESULTS: We isolated 1955 H. pylori strains. The resistance rates of H. pylori to amoxicillin, levofloxacin, clarithromycin, and metronidazole were 0.9%, 42.4%, 40.4%, and 94.2%, respectively. Only five tetracycline-resistant and one furazolidone-resistant strain were identified. Overall, 3.3% of the strains were sensitive to all six antibiotics. Multidrug-resistant strains accounted for 22.9%, of which less than 20% were from Wuzhong. Strains isolated from women and patients with nonulcerative disease had higher rates of resistance to levofloxacin and clarithromycin. Higher rates of resistance to metronidazole, levofloxacin, and clarithromycin were observed in the older age group than in the younger age group. The kappa coefficients of phenotypic resistance and genotypic resistance for levofloxacin and clarithromycin were 0.830 and 0.809, respectively, whereas the remaining antibiotics showed poor agreement. CONCLUSION: H. pylori antibiotic resistance is severe in Ningxia. Therefore, furazolidone, amoxicillin, and tetracycline are better choices for the empirical therapy of H. pylori infection in this region. Host sex, age, and the presence of ulcerative diseases may affect antibiotic resistance of the bacteria. Personalized therapy based on genetic testing for levofloxacin and clarithromycin resistance may be a future direction for the eradication therapy of H. pylori infection in Ningxia.

Overexpression of TaSIM provides increased drought stress tolerance in transgenic Arabidopsis.

Drought is the most serious meteorological disaster affecting wheat production. Members of the R2R3-MYB gene subfamily play a crucial role in the regulation of the wheat drought stress response. In this study, the function of polyethylene glycol (PEG)-induced expression of the wheat R2R3-MYB gene TaSIM in response to drought stress was characterized. ß-Glucuronidase (GUS) histochemical staining revealed that the TaSIM promoter can drive the expression of the GUS gene in the flowers, roots, stems and rosette leaves. Moreover, TaSIM was expressed in the stamens, pistils, roots, stems and leaves of wheat. The TaSIM promoter contains a known stress-responsive cis-acting element and is inducible by stress, PEG and abscisic acid (ABA). Under drought stress, compared with wild-type (WT) Arabidopsis, transgenic Arabidopsis overexpressing TaSIM presented significantly lower leaf water loss rates and increased survival. Moreover, the content of soluble sugars and proline and the expression of stress-related genes (RD29A and RD22) in transgenic Arabidopsis overexpressing TaSIM were higher than those in WT Arabidopsis under drought stress. Our results indicate that TaSIM plays a positive role in the drought stress response and can be used as a candidate gene for the genetic engineering of wheat drought resistance.

The wheat salinity-induced R2R3-MYB transcription factor TaSIM confers salt stress tolerance in Arabidopsis thaliana.

MYB transcription factors are a large family of proteins involved in plant development and responses to stress. In this study, the wheat salinity-induced R2R3-MYB transcription factor TaSIM was functionally characterized, with a focus on its role in salt stress tolerance. TaSIM protein enters the nucleus and binds to the MYB-binding site II motif. Expression analysis revealed that TaSIM was induced by drought, high salinity, low temperature, and abscisic acid treatment. Overexpression of TaSIM improved salt stress tolerance in transgenic plants. Furthermore, the transcript levels of genes involved in abscisic acid (ABA)-dependent (RD22) and ABA-independent (RD29A) signaling were higher in TaSIM-overexpressing plants than in the wild type. These results suggest that TaSIM positively modulates salt stress tolerance and has potential applications in molecular breeding to enhance salt tolerance in crops.

Pulmonary hypertension risk in patients with hemoglobin h disease: low incidence and absence of correlation with splenectomy.

Pulmonary hypertension (PHT) is a common complication for patients with ß thalassemia intermediate (TI), especially splenectomized patients. However, the frequency and risk factors of PHT in patients with hemoglobin H (HbH) disease is unknown. The purpose of this study was to identify the prevalence of PHT risk manifested as tricuspid regurgitant jet velocity (TRV) ≥2.5 m/s in patients with HbH disease and its correlation with splenectomy. One hundred and ninety-eight patients with HbH disease who visited the 303rd Hospital of the People's Liberation Army (Nanning, China) were investigated. Thirteen subjects (6.5%) were diagnosed as having a risk of PHT. Regression analyses showed that the prevalence of PHT risk was correlated only with age (r = 0.195, p = 0.006) and not with splenectomy. The risk of PHT in patients older than 35 years was 5.7 times (range 1.8-18.6) greater than that for patients younger than 35 years. For splenectomized patients compared to those with HbH disease, patients with TI had a higher frequency of PHT risk, higher nucleated red blood cell counts (46.03 ± 41.11 × 10(9)/l vs. 0.18 ± 1.19 × 10(9)/l, p < 0.001) and a higher platelet counts (837.6 ± 178.9 × 10(9)/l vs. 506.7 ± 146.2 × 10(9)/l, p < 0.001). PHT risk is low in patients with HbH disease and does not correlate with splenectomy. Patients older than 35 years should be monitored regularly.

G-quadruplex guanosine gels and single walled carbon nanotubes.

Solubilization of single walled carbon nanotubes (SWNTs) in aqueous gel phases formed by reversible, G-quadruplex self-assembly of guanosine monophosphate (GMP) alone or with guanosine (Guo) is described. Unlike other media and methods for aqueous solubilization of SWNTs, the guanosine gels ("G-gels") are found to readily disperse high (>mg/mL) concentrations of individual rather than bundled SWNTs. SWNT dispersions in GMP alone precipitate in several hours and re-form upon shaking; however, dispersions in the binary GMP/Guo gels are indefinitely stable. Increasing GMP or KCl concentration in the binary gels increased the relative abundance of large diameter and semi-conducting SWNTs. Different gel compositions also displayed different selectivities toward SWNTs of different chiralities. These results indicate a strong connection between the self-assembled G-gels and the dimensions and structures of the SWNTs that they solubilize.

The Gma-miR394a/GmFBX176 module is involved in regulating the soybean (Glycine max L.) response to drought stress.

Drought seriously affects the yield and quality of soybean. Previous studies have shown that the gma-miR394a/GmFBX176 module regulates the response of Arabidopsis to drought stress. However, whether the gma-miR394a/GmFBX176 module is involved in the regulation of the soybean drought stress response remains unclear. Here, the function of the gma-miR394a/GmFBX176 module in the soybean drought stress response was evaluated. In soybean hairy roots, drought stress induced the transcription of gma-miR394a and inhibited the transcription of GmFBX176. GUS histochemical staining showed that transgenic GmFBX176p:GUS soybean hairy root staining was weak and that GUS transcript levels decreased under drought stress. A transient expression experiment in tobacco showed that gma-miR394a inhibited GmFBX176 transcription. Under drought stress, composite soybean plants overexpressing gma-miR394a showed increased drought resistance compared with control K599 composite soybean plants (K599); their survival rate and peroxidase activity were higher than those of K599, and their malondialdehyde content was lower. In contrast, composite soybean plants overexpressing GmFBX176m3 (gma-miR394a complement site mutation) presented lower drought resistance than K599 plants. Transcriptomic sequencing showed that the gma-miR394a/GmFBX176 module affected the transcript levels of stress response genes and transcription factors. These results indicate that the gma-miR394a/GmFBX176 module can be used to improve the drought resistance of soybean.

Endoscopic Ruler for varix size measurement: A multicenter pilot study.

BACKGROUND: We invented Endoscopic Ruler, a new endoscopic device to measure the size of varices in patients with cirrhosis and portal hypertension. AIM: To assess the feasibility and safety of Endoscopic Ruler, and evaluate the agreement on identifying large oesophageal varices (OV) between Endoscopic Ruler and the endoscopists, as well as the interobserver agreement on diagnosing large OV using Endoscopic Ruler. METHODS: We prospectively and consecutively enrolled patients with cirrhosis from 11 hospitals, all of whom got esophagogastroduodenoscopy (EGD) with Endoscopic Ruler. The primary study outcome was a successful measurement of the size of varices using Endoscopic Ruler. The secondary outcomes included adverse events, operation time, the agreement of identifying large OV between the objective measurement of Endoscopic Ruler and the empirical reading of endoscopists, together with the interobserver agreement on diagnosing large OV by Endoscopic Ruler. RESULTS: From November 2020 to April 2022, a total of 120 eligible patients with cirrhosis were recruited and all of them underwent EGD examinations with Endoscopic Ruler successfully without any adverse event. The median operation time of Endoscopic Ruler was 3.00 min [interquartile range (IQR): 3.00 min]. The kappa value between Endoscopic Ruler and the endoscopists while detecting large OV was 0.52, demonstrating a moderate agreement. The kappa value for diagnosing large OV using Endoscopic Ruler among the six independent observers was 0.77, demonstrating a substantial agreement. CONCLUSION: The data demonstrates that Endoscopic Ruler is feasible and safe for measuring the size of varices in patients with cirrhosis and portal hypertension. Endoscopic Ruler is potential to promote the clinical practice of the two-grade classification system of OV.

Enzymatic conversion of CO(2) to bicarbonate in functionalized mesoporous silica.

We report here a concept converting carbon dioxide to biocarbonate in a biomimetic nanoconfiguration. Carbonic anhydrase (CA), the fastest enzyme that can covert carbon dioxide to bicarbonate, can be spontaneously entrapped in carboxylic acid group-functionalized mesoporous silica (HOOC-FMS) with super-high loading density (up to 0.5 mg of protein/mg of FMS) in sharp contrast to normal porous silica. The binding of CA to HOOC-FMS resulted in a partial conformational change comparing to the enzyme free in solution, but it can be overcome with increased protein loading density. The higher the protein loading density, the less conformational change, hence the higher enzymatic activity and the higher enzyme immobilization efficiency (up to >60%). The released enzyme still displayed the native conformational structure and the same high enzymatic activity as that prior to the enzyme entrapment, indicating that the conformational change resulted from the electrostatic interaction of CA with HOOC-FMS was not permanent. This work may provide a new approach converting carbon dioxide to biocarbonate that can be integrated with the other part of biosynthesis process for the assimilation of carbon dioxide.

The sea-island cotton GbTCP4 transcription factor positively regulates drought and salt stress responses.

TCP transcription factors play important regulatory roles in plant growth and development; however, their function in response to salt and drought stress in sea-island cotton (Gossypium barbadense) is unknown. Here, GbTCP4 expression was induced by abscisic acid (ABA), drought, and NaCl treatments. Under drought stress, compared to wild-type (WT) Arabidopsis, transgenic GbTCP4-overexpressing Arabidopsis showed increased seed germination rate, root length and survival rate; additionally, it was ABA-insensitive at the germination stage but ABA-sensitive at the seedling stage, showing reduced stomatal opening and ABA enrichment. Under salt stress, compared to WT Arabidopsis, transgenic GbTCP4-overexpressing Arabidopsis showed greater root length, survival rate, and SPAD value and lower malondialdehyde (MDA) content. Conversely, under drought or salt stress, virus-induced gene-silenced GbTCP4 cotton showed decreased root length, area and volume and increased MDA content and sensitivity to drought and salt stress compared with control cotton. RNA-seq and quantitative real-time PCR analyses showed that GbTCP4 affected the transcription levels of genes across multiple abiotic stress-related metabolic pathways. Furthermore, GbTCP4 activated the transcription of GbUVR8 and GbbHLH130 by binding to their promoters. These results suggest that GbTCP4 positively regulates drought and salt stress responses and is a suitable candidate gene for improving plant drought and salt tolerance.

Study on population structure of kiwifruit and GWAS for hairiness character.

BACKGROUND: A total of 74,936 SNPs were employed to carry out population structure and genome-wide association studies and post-GWAS for hairiness character of the fifty-six samples including thirty-six Actinidia chinensis var. deliciosa, eighteen A. chinensis var. chinensis, and two A. polygama in the light of morphological observations. RESULTS: The percentage of heterozygous sites of A. chinensis var. deliciosa is higher than that of A. chinensis var. chinensis, which could be one of the reasons for A. chinensis var. deliciosa high disease resistance. Fifty-six samples were divided into two subgroups, in which the genetic distance, ranged from 0.17 to 0.99, according to their genetic divergence. Analysis of molecular variance shows that the frequency of genetic variations within the population is 83.53% and 16.47% between populations. Fst between the two populations is 0.14, and Nm is 1.60. Set at α ≤ 0.05, a total of 327 SNPs and 260 haplotypes were related to the hairiness character. A total of 246 proteins were annotated using GO and KEGG analyses, which indicated the membrane-related genes and stress-resistant metabolic pathways are related to the hairiness character of leaves, stems, and peels of kiwifruit. Protein interaction analysis showed that DNA-directed RNA polymerase was an important node protein that interacted with many proteins. CONCLUSIONS: The genetic basic in the fifty-six genotypes was rich. The results of clustering and morphological observations are not completely consistent, indicating the hairiness character play an important role in the classification of kiwifruit, in which two A. polygama were clustered together with those of A. chinensis var. chinensis. Phylogeny and haplotype analysis showed that the evolution of A. chinensis var. chinensis is later than that of A. chinensis var. deliciosa in A. chinesis. The loss of hairiness character on leaves, stems and peels of A. chinensis var. chinensis compare with A. chinensis var. deliciosa, which is also the result of its poor resistance.

Heterologous overexpression of the GbTCP5 gene increased root hair length, root hair and stem trichome density, and lignin content in transgenic Arabidopsis.

Teosinte branched1/cycloidea/proliferating cell factor1 (TCP) is a plant-specific protein family member involved in plant growth and development. However, the functions of most members of the cotton TCP family are unknown. In this study, the GbTCP5 gene encodes a sea-island cotton class II TCP CIN subclass transcription factor. The GbTCP5 transcription factor is located in the nucleus, has transcriptional activation activity, and can bind to TCP II cis-acting elements. GbTCP5 was widely expressed in tissues with the highest transcript level in the calyx. GbTCP5 is expressed at different developmental stages of the fiber and has significantly high transcriptional level expression in the fibers at 20, 30 and 35 days post anthesis (DPA). Heterologous overexpression of the GbTCP5 gene increased root hair length, root hair and stem trichome density, and stem lignin content in transgenic Arabidopsis compared to the wild type (WT). GbTCP5 binds the promoters of the GL3, EGL3, CPC, MYB46, LBD30, CesA4, VND7, CCOMT1, and CAD5 genes to upregulate their expression. Moreover, the homologous genes of these genes are expressed in the fibers of different developmental stages of the sea-island cotton fiber. These results indicate that GbTCP5 regulates root hair development and secondary wall formation in Arabidopsis and may be a candidate gene for improving cotton fiber quality.

Heterologous Expression of GbTCP4, a Class II TCP Transcription Factor, Regulates Trichome Formation and Root Hair Development in Arabidopsis.

Two class I family teosinte branched1/cycloidea/proliferating cell factor1 (TCP) proteins from allotetraploid cotton are involved in cotton fiber cell differentiation and elongation and root hair development. However, the biological function of most class II TCP proteins is unclear. This study sought to reveal the characteristics and functions of the sea-island cotton class II TCP gene GbTCP4 by biochemical, genetic, and molecular biology methods. GbTCP4 protein localizes to nuclei, binding two types of TCP-binding cis-acting elements, including the one in its promoter. Expression pattern analysis revealed that GbTCP4 is widely expressed in tissues, with the highest level in flowers. GbTCP4 is expressed at different fiber development stages and has high transcription in fibers beginning at 5 days post anthesis (DPA). GbTCP4 overexpression increases primary root hair length and density and leaf and stem trichomes in transgenic Arabidopsis relative to wild-type plants (WT). GbTCP4 binds directly to the CAPRICE (CPC) promoter, increasing CPC transcript levels in roots and reducing them in leaves. Compared with WT plants, lignin content in the stems of transgenic Arabidopsis overexpressing GbTCP4 increased, and AtCAD5 gene transcript levels increased. These results suggest that GbTCP4 regulates trichome formation and root hair development in Arabidopsis and may be a candidate gene for regulating cotton fiber elongation.

Overexpression of soybean miR169c confers increased drought stress sensitivity in transgenic Arabidopsis thaliana.

The miR169 family, a large-scale microRNA gene family conserved in plants, is involved in stress responses, although how soybean miR169 functions in response to drought stress remains unclear. We show that gma-miR169c exerts a negative regulatory role in the response to drought stress by inhibiting the expression of its target gene, nuclear factor Y-A (NF-YA). A real-time RT-PCR analysis indicated that gma-miR169c is widely expressed in soybean tissues and induced by polyethylene glycol (PEG), high salt, cold stress and abscisic acid (ABA). Histochemical ß-glucuronidase (GUS) staining showed that the gma-miR169c promoter drives GUS reporter gene expression in various transgenic Arabidopsis tissues, and the stress-induced pattern was confirmed in transgenic Arabidopsis and transgenic soybean hairy roots. Arabidopsis overexpressing gma-miR169c is more sensitive to drought stress, with reduced survival, accelerated leaf water loss, and shorter root length than wild-type plants. We identified a precise cleavage site for 10 gma-miR169c targets and found reduced transcript levels of the AtNFYA1 and AtNFYA5 transcription factors in gma-miR169c-overexpressing Arabidopsis and reduced expression of the stress response genes AtRD29A, AtRD22, AtGSTU25 and AtCOR15A. These results indicate that gma-miR169c plays a negative regulatory role in drought stress and is a candidate miRNA for improving plant drought adaptation.

Tunable thermoassociation of binary guanosine gels.

It is well-known that aqueous solutions of individual guanosine compounds can form gels through reversible self-assembly. Typically, gelation is favored at low temperature and acidic pH. We have discovered that binary mixtures of 5'-guanosine monophosphate (GMP) and guanosine (Guo) can form stable gels at neutral pH over a temperature range that can be tuned by varying the relative proportions of the hydrophobic Guo and the hydrophilic GMP in the mixture. Gelation was studied over the temperature range of 5-40 degrees C or 60 degrees C at pH 7.2 using visual detection, circular dichroism (CD) spectroscopy, and CD thermal melt experiments. Solutions with high GMP/Guo ratios behaved similar to solutions of GMP alone while solutions with low GMP/Guo formed firm gels across the entire temperature range. Most interesting were solutions between these two extremes, which were found to exhibit thermoassociative behavior; these solutions are liquid at refrigerator temperature and undergo sharp transitions to a gel only at higher temperatures. Increasing the GMP/Guo ratio and increasing the total concentration of guanosine compounds shifted the onset of gelation to higher temperatures (ranging from 20 to 40 degrees C), narrowed the temperature range of the gel phase, and sharpened the reversible phase transitions. The combination of self-assembly, reversibility, and tunability over biologically relevant temperature ranges and pH offers exciting possibilities for these simple and inexpensive materials in medical, biological, analytical, and nanotechnological applications.

Delivery of MicroRNA-10b with Polylysine Nanoparticles for Inhibition of Breast Cancer Cell Wound Healing.

Recent studies revealed that micro RNA-10b (mir-10b) is highly expressed in metastatic breast cancer cells and positively regulates breast cancer cell migration and invasion through inhibition of HOXD10 target synthesis. In this study we designed anti-mir-10b molecules and combined them with poly L-lysine (PLL) to test the delivery effectiveness. An RNA molecule sequence exactly matching the mature mir-10b minor antisense showed strong inhibition when mixed with PLL in a wound-healing assay with human breast cell line MDA-MB-231. The resulting PLL-RNA nanoparticles delivered the anti-microRNA molecules into cytoplasm of breast cancer cells in a concentration-dependent manner that displayed sustainable effectiveness.

[Observation of pathological morphology on nervous system of rat with maximum dose sophoridine in chronic toxic test].

BACKGROUND & OBJECTIVE: Sophoridine is a new anticancer drug with noticeable antitumor action and lower toxicity. No marked influence on bone marrow was found till now. The main toxicity is presented in nervous system. This study was to observe the morphological changes of the nervous system of the rats, which were treated with maximum dose of sorphoridine for a long time. METHODS: 30 rats,half of male when and half of female, were randomly divided into experimental group and control group. In the experimental group,rats were treated with maximum dose of sorphoridine [32 mg x(kg x d)(-1) ip, qd] for 60 days. In control group, rats were treated with the same volume of saline everyday for 60 days. The rats in both groups were killed at 20 d, 40 d, 60 d, and 75 d, respectively. The brain and spinal cord were taken out and made into pathological slices, which were stained by HE stain and special stain, sach as Nissel's body stain, glial fibrillary stain and myelin sheath stain. The differences in morphology between the two groups was observed. RESULTS: No pathological change was found in rats' cerebral cortex,internal capsul, striated body, hippocampus, substantia nigra,and spinal cord when the rats' were treated with sophoridine 32 mg x(kg x d)(-1) ip for 20 d, 40 d, 60 d. In the rats who had presented nervous system syndrome repeatedly or died for convulsion, or the rats who were killed in convalescence period (15 d after final administration), there was no pathological change either. CONCLUSION: No pathological changes and delayed changes in the nervous tissues were found when the rats were given maximum dose of Sophoridine continuously for 60 d. Our study showed that the syndrome of nervous system caused by Sophoridine is functional and stimulational, and can be recovered,and there is no any delayed change and sequela.