Evaluating the effectiveness and safety of acupuncture on serum uric acid in asymptomatic hyperuricemia population: a randomized controlled clinical trial study protocol.

Background: The clinical dangers of asymptomatic hyperuricemia to human health have become increasingly prominent over the past 20 years. Previous studies have shown the potential benefits of acupuncture on uric acid levels in the body. However, definitive evidence is lacking. Our objective is to evaluate the efficacy and safety of acupuncture on serum uric acid (SUA) in individuals with asymptomatic hyperuricemia. Methods: This is a randomized, single-blind, sham-controlled trial. A total of 180 eligible patients with asymptomatic hyperuricemia will be recruited at three hospitals in China. Patients will be randomly assigned in a 1:1 ratio to receive 16 sessions of manual acupuncture or sham acupuncture for 8 weeks. Patients will be followed up for 12 weeks. The primary outcome will be the change in SUA levels at week 8 after randomization. Secondary outcomes will include dynamic changes in SUA levels, efficacy rates, proportion of gout flare, body weight, and acute medication intake. The MGH Acupuncture Sensation Scale and adverse events related to acupuncture will be measured after each treatment. A blinding assessment will be performed on patients who receive at least one session of acupuncture. Data analyses will be performed on a full analysis set and a per-protocol set. Ethics and dissemination: Ethics approval has been obtained from the Clinical Trial Ethics Committee of Tongji Medical College, Huazhong University of Science and Technology (approval no. 2021-S135). Written informed consent will be obtained from enrolled patients. The findings will be disseminated in a peer-reviewed journal. Clinical trial registration: ClinicalTrials.gov identifier, NCT05406830.

[Effects of Immune Cells on Luteal Development and Regression in the Mammalian Ovary].

Corpus luteum,an important endocrine tissue in mammalian ovary,plays a role in the regulation of reproductive cycle and the establishment of early pregnancy.While studying the luteal development and its molecular regulation,we have discovered a variety of immune cells,such as T lymphocytes,macrophages,neutrophils,and eosinophils,in the corpus luteum.These immune cells accumulate and support luteal angiogenesis and progesterone production during the luteal development,thus participating in the regulation of luteal functions.In luteal regression,prostaglandin F2 can stimulate the production of inflammatory cytokines and chemokines,which help immune cells enter the corpus luteum and enhance the decomposition of corpus luteum through inflammatory reactions.According to our research achievements,we reviewed the roles of different types of immune cells in the development and degradation of mammalian luteal functions,aiming to further understand the biology of corpus luteum and provide a reference for the clinical manipulation of luteal functions.

Sigma­1 receptor overexpression promotes proliferation and ameliorates cell apoptosis in ß­cells.

Sigma­1 receptor (Sig­1R) is a class of orphan receptors, the potential role of which in pancreatic islet cells remains poorly understood. The present study aimed to investigate the role of Sig­1R in islet ß­cell proliferation and examine the effects of Sig­1R on islet ß­cell injury under lipotoxic conditions. Sig­1R­overexpressing MIN6 cells were generated by lentiviral vector transfection. The effect of Sig­1R overexpression on cell proliferation detected by EdU staining, cell cycle progression by propidium iodide (PI), apoptosis by Annexin V­APC/PI, mitochondrial membrane potential by Mitolite Red and cytoplasmic Ca2+ levelsby Fura­2/AM in islet ß­cells were measured by flow cytometry. Western blot analysis was used to measure protein expression levels of endoplasmic reticulum (ER) stress markers glucose­regulated protein 78 and C/EBP homologous protein, mitochondrial apoptotic proteins Bcl­2­associated X and Bcl­2 and cytochrome c. In addition, ATP levels and insulin secretion were separately measured using ATP Assay and mouse insulin ELISA. Mitochondria­associated ER membrane (MAM) structures in MIN6 cells were then detected using transmission electron microscopy. Protein disulfide isomerase expression and possible colocalization between inositol 1,4,5­trisphosphate receptor and voltage­dependent anion channel 1 were examined using immunofluorescence. Sig­1R overexpression was found to promote ß­cell proliferation by accelerating cell cycle progression. Furthermore, Sig­1R overexpression ameliorated the apoptosis rate whilst impairing insulin secretion induced by palmitic acid by relieving ER stress and mitochondrial dysfunction in MIN6 cells. Sig­1R overexpression also promoted Ca2+ transport between mitochondria and ER by increasing the quantity of ER adjacent to mitochondria in the 50­nm range. It was concluded that Sig­1R overexpression conferred protective effects on ß­cells against lipotoxicity as a result of the promotion of cell proliferation and inhibition of ER stress and oxidative stress, by regulating the structure of MAM.

Facile Molecular Weight Determination of Polymer Brushes Grafted from One-Dimensional Diffraction Grating by SI-ATRP Using Reflective Laser System.

Gelatin was immobilized selectively on the amide groups-modified bottom of a trench array of a photoresist template with 2 µm resolution by the ethyl(dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide reaction. The gelatin-immobilized line array was brominated to generate a macroinitiator for atom transfer radical polymerization. Poly(methacrylic acid) (PMAA) brushes were grafted from the macroinitiator layer as line arrays of one-dimensional diffraction gratings (DGs) for various grafting polymerization times. A laser beam system was employed to analyze the optical feature with a characteristic diffraction effect of the PMAA DGs at a 45° incident angle along the transverse magnetic and transverse electric polarization. The growth of the PMAA brush lines increased both their heights and widths, leading to a change in the reflective diffraction intensity. The PMAA brushes under various grafting polymerization times were cleaved from the substrate by digestion of gelatin with trypsin, and their molecular weights were obtained by gel permeation chromatography. The change degree of the diffraction intensity varied linearly with the molecular weight of the PMAA brushes over a wide range, from 135 to 1475 kDa, with high correlation coefficients. Molecular weight determination of polymer brushes using the reflective diffraction intensity provides a simple method to monitor their growth in real time without polymer brush cleavage.

HIF-1α Protects Granulosa Cells From Hypoxia-Induced Apoptosis During Follicular Development by Inducing Autophagy.

Owing to the avascular structure of the ovarian follicle, proliferation of granulosa cells (GCs) and development of follicles occur under hypoxia, which is obviously different from the cell survival requirements of most mammalian cells. We hypothesized that autophagy may exert an inhibitory effect on GC apoptosis. To decipher the underlying mechanism, we constructed a rat follicular development model using pregnant mare serum gonadotropin and a cell culture experiment in hypoxic conditions (3% O2). The present results showed that the autophagy level was obviously increased and was accompanied by the concomitant elevation of hypoxia inducible factor (HIF)-1α and BNIP3 (Bcl-2/adenovirus E1B 19kDa-interacting protein 3) in GCs during follicular development. The levels of Bax (Bcl2-associated X) and Bcl-2 (B-cell lymphoma-2) were increased, while the activation of caspase-3 exhibited no obvious changes during follicular development. However, inhibition of HIF-1α attenuated the increase in Bcl-2 and promoted the increase in Bax and cleaved caspase-3. Furthermore, we observed the downregulation of BNIP3 and the decrease in autophagy after treatment with a specific HIF-1α activity inhibitor (echinomycin), indicating that HIF-1α/BNIP3 was involved in autophagy regulation in GCs in vivo. In an in vitro study, we also found that hypoxia did not obviously promote GC apoptosis, while it significantly enhanced the activation of HIF-1α/BNIP3 and the induction of autophagy. Expectedly, this effect could be reversed by 3-methyladenine (3-MA) treatment. Taken together, these findings demonstrated that hypoxia drives the activation of HIF-1α/BNIP3 signaling, which induces an increase in autophagy, protecting GC from apoptosis during follicular development.

Expression Regulation and Physiological Role of Transcription Factor FOXO3a During Ovarian Follicular Development.

In mammals, developing ovarian follicles transform from primordial follicles to primary follicles, secondary follicles, and mature follicles, accompanied by changes in follicular secretory functions. FoxO3a is a member of the forkhead transcription factor family (FoxO), which plays an important role in the cell cycle, DNA damage repair, apoptosis, oxidative stress, and energy metabolism. Recent studies have shown that FOXO3a is involved in the physiological regulation of follicular development and pathological progression of related ovarian diseases, which will provide useful concepts and strategies for retarding ovarian aging, prolonging the ovarian life span, and treating ovarian diseases. Therefore, the regulation of FOXO3a expression, as well as the physiological contribution during ovarian follicular development are detailed in this paper, presenting an important reference for the further study of ovarian biology.

Electrorheological Sensor Encapsulating Microsphere Media for Plague Diagnosis with Rapid Visualization.

Plague is a disease infected by an etiological agent, which is transmitted from fleas to a variety of wildlife rodents. Therefore, rapid diagnosis of plague on-site in the field is important. Polystyrene microspheres (SMs) of 2.2 µm diameter were synthesized by emulsion polymerization to adsorb magnetic nanoparticles (FNs), resulting in core-/shell-structured microspheres that generate a significant contrast in relative permittivities between SMs and FNs. Electrorheological displays (EDs) consisting of two indium tin oxide glasses with spacers were constructed to contain core-/shell-structured SM/FN (SM@FN) solutions for observing their transmittance change. The ED encapsulating dispersed SM@FN solution exhibited an opaque state because light was scattered significantly without the application of an alternating electric field (AEF). In the presence of an AEF, the particle chaining behavior results in enhancement of the transmittance of ED. At a specific frequency, the so-called characteristic frequency (Fc), the transmittance reaches a maximum. Fc could be used as an indicator to mark the shell materials. The antibody of Yersinia pestis (ab-Yp) was coated onto the SM@FN as a biosensing medium. The Fc of ab-Yp-modified microspheres shifted from 200 to 750 kHz with antigen coupling of Y. pestis antigen (ag-Yp). In the absence of fluorescence labeling, the large change in ED transmittance could be visualized during the Y. pestis detection. The limit of detection and the limit of quantification were ∼30 and ∼40 ng/µL, respectively, obtained within 30 s according to the highest transmittance of ED under the AEF at 750 kHz. Y. pestis detection was not affected by Escherichia coli and Staphylococcus aureus significantly. Compared with other common immunoassays, including the secondary immunochemical or enzyme-linked steps, this simple electrorheological sensor with high sensitivity and selectivity could be a candidate for on-site plague diagnosis.

Coordination between Surface Lattice Resonances of Poly(glycidyl Methacrylate) Line Array and Surface Plasmon Resonances of CdS Quantum on Silicon Surface.

In this work, a unique hybrid system is proposed for one-dimensional gratings comprising of poly(glycidyl methacrylate) (PGMA) brushes and CdS quantum dots (CQDs). Generally, the emission of QDs is too weak to be observed in a dry state. Plasmonic resonances of the grating structures can be used to enhance the light emission or absorption of CQDs. The interaction between PGMA plasmonic nanostructures and inorganic CQDs plays a crucial role in engineering the light harvest, notably for optoelectronic applications. Extinction measurements of the hybrid system consisting of a PGMA grating and CQDs are reported. We designed one-dimensional gratings with various resolutions to tune the absorptance peaks of grating. PGMA grating grafted from a 1.5 µm resolution of trench arrays of photoresist exhibited absorptance peak at 395 nm, close to the absorption peak of CQDs, resulting in the photoluminescence enhancement of CQDs on the grating due to high charge carriers' recombination rate. Generally, the emission of quantum dots occurs under irradiation at characteristic wavelengths. Immobilizing QDs on the grating facilitates the emission of QDs under irradiation of full-wavelength light. Furthermore, the PGMA gratings with CQDs were immersed in various solvents to change the geometries resulting the shift of absorptance peak of grating. The proposed method could be applied for sensing the nature of the surrounding media and vice versa, as well as for various media of solvents.

Surface lattice resonance of line array of poly (glycidyl methacrylate) with CdS quantum dots for label-free biosensing.

One dimensional plasmonic grating is a kind of resonant electromagnetic wave absorber with a characteristic wavelength. This study focusses on one-dimensional plasmonic grating consisting of poly (glycidyl methacrylate) (PGMA) brushes and CdS quantum dots (CdQDs) fabrication and PGMA chains grafted on a primary substrate in a line array continued by the immobilization of biotin-modified CdQDs. PGMA brush line array (PBLA) of plasmonic grating exhibited an absorptance at 441 nm while at the same time, CdQDs immobilized with PBLA showed characteristic absorbance at 396 nm. The blue-shift from 441 nm matches the absorbance peak of biotin-modified CdQDs resulting in the enhancement of photoluminescence emission of CdQDs. With streptavidin incubation to assemble CdQDs at 50 nM, the significant decrease in grating height resulted in the red-shift of the absorbance peak to 536 nm. Due to the deviation in absorbance, the intensity of the PL emission decreased gradually with the increase in concentration of streptavidin. In addition, our results showed that streptavidin incubation altered the color reflected from the surface due to effective changes in the refractive index of the layer as well. The limit of detection of the grating for streptavidin detection was determined to be 50 nM. Thus, PBLA-CdQD has the potential to act as a highly-sensitive, label-free optical biosensor.

Rapid and sensitive detection of Yersinia pestis by lateral-flow assay in simulated clinical samples.

BACKGROUND: Yersinia pestis is a contributing agent to the epidemic disease, plague, which killed an estimated 200 million people during historical times. In this study, a rapid, cheap, sensitive, and specific technique, the lateral flow assay (F1 strips), has been successfully developed to detect this pathogen, by using paired monoclonal antibodies (MAbs) against Y. pestis capsule like fraction 1 (F1) protein. Compared with the polyclonal antibody (PAb) based F1 strips, the Mab-based F1 strips have a remarkable increased detection limitation (10 to 100 folds). Furthermore, besides the limitation and specificity evaluation, the application of this F1 strip on simulated clinical samples indicate the LFA can be a good candidate to detect plague. METHODS: Recombinant F1 antigen was expressed and purified from a series of works. The various anti-F1 monoclonal antibodies generated from hybridoma cells were screened with the ELISA technique. To evaluate the feasibility of this Y. pestis F1 test strip, the F1 protein/Y. pestis was spiked into simulated clinical samples such as human serum, mouse bronchoalveolar lavage fluids, and mouse blood to mimic natural infection status. Additionally, this technique was applied to detect the Y. pestis in the environment-captured rats, to evaluate the practical usefulness of the strips. RESULTS: By using this MAb-based-LFA technique, 4 ng/ml of recombinant F1-protein and 103 CFU/ml of Y. pestis could be detected in less than 10 mins, which is at least 10-folds than that of the PAb format. On the other hand, although various Yersinia strains were applied to the strips, only Y. pestis strain showed a positive result; all other Yersinia species did not produce a positive signal, indicating the high efficiency and specificity of the MAb-based F1-strips. CONCLUSION: Based on our findings, we suggest that the MAb-format-LFA will be valuable as a diagnostic tool for the detection of Y. pestis. This report shows that the F1 strip is sufficient to support not only the detection of plague in simulated clinical samples, but also it may be a good candidate to meet the epidemiological surveillance during an outbreak of the biological warfare.

Suspension bead array of the single-stranded multiplex polymerase chain reaction amplicons for enhanced identification and quantification of multiple pathogens.

Rapid identification of single and multiple infectious agents is vital in clinical settings and during biothreat attack. This study assesses the assay of single-stranded multiplex polymerase chain reaction (PCR) amplicons by suspension bead array (SSMP-SBA) for multiple pathogens identification in a single-tube reaction. A 15-plex assay for identification of 11 highly infectious pathogens was developed to evaluate the performance of SSMP-SBA. Pathogen-specific amplicons were obtained by sequential amplification of genomic DNAs using gene-specific primers tagged with artificial unique sequences and unique primers of which the reverse primer was modified by biotin and phosphorothioate. The SSMP products generated by T7 exonuclease-mediated DNA hydrolysis were hybridized to 15 sets of beads coupled with gene-specific and control oligonucleotide probes for pathogen identification and quantification by flow cytometry. This method was validated via assessment of 57 reference strains and one clinical bacterial isolate. All 11 pathogens can be detected by the 15-plex SSMP-SBA assay, and this design significantly enhanced the signal-to-noise ratio and improved the assay performance. This assay achieves similar sensitivity to our in-house real-time PCR system with the limit of detection equivalent to 5-100 genome copies and a linear dynamic range crossing three to five logs. In the validation assay, a 100% accuracy rate was achieved when the pathogens were among the target species. Notably, the species of pathogens were accurately identified from the samples with multiple infections. SSMP-SBA presents superior performance with multiplexing capability in a single-tube reaction and provides a new approach for detection and species identification of multiple pathogen infections.

In vitro and in vivo anticancer activity evaluation of ursolic acid derivatives.

Twenty-three ursolic acid (1) derivatives 2-24 (ten novel compounds 8-10, 14-17 and 22-24) modified at the C-3 and the C-28 positions were synthesized, and their structures were confirmed by IR, (1)H NMR, MS, and elemental analysis. The single crystals of compounds 15 and 17 were obtained. The cytotoxic activity of the derivatives was evaluated against HepG2, BGC-823, SH-SY5Y, HeLa and HELF cells by the MTT assay. The induction of apoptosis and affects on the cell cycle distribution with compound 14 were assessed by fluorescence microscopy, flow cytometry and the activity of caspase-3 in HepG2 cells. Compounds 14-17 had more significant antiproliferative ability against the four cancer cell lines and low cytotoxicity to human embryonic lung fibroblast cells (HELF). Compounds 11, 14-16, 21 and 23 were particularly active against HepG2 cell growth. Compound 14 was selected to investigate cell apoptosis and cell cycle distribution. Flow cytometric analysis and morphologic changes of the cell exhibited that treatment of HepG2 cells with compound 14 led to cell apoptosis accompanied by cell cycle arrest at the S phase in a dose-dependent manner. Furthermore, the activity of the caspase-3 enzyme was increased in the treated cells. In vivo studies using H22 xenografts in Kunming mice were conducted with compound 14 at doses of 50, 100 and 150 mg/kg body weight. The results revealed that the medium dosage group (100 mg/kg) showed significant anticancer activity (45.6 ± 4.3%) compared to the control group.

[Transcriptional regulation of cytochrome P450 3A4 by four kinds of traditional Chinese medicines].

OBJECTIVE: To screen a group of traditional Chinese medicines with effect on pregnane X receptor (PXR)-mediated transcription regulation of P450 3A4 (CYP3A4); and to study whether they can induce the expression of CYP3A4 with a dose, time-dependent manner. METHOD: Transient cotransfection reporter gene assays were performed with pCI-hPXR-neo, pGL3-CYP3A4-Luc and beta-galactosidase expression plasmid in HepG2 cells. RESULT: Rhizoma Curcumae, Atractylodes lancea, A. macrocaphala and Poria cocos could induce transcriptional expression of CYP3A4. In the dose-effect study, 24 h after induction, 500 mg x L(-1) Rhizoma Curcumae, A. lancea, A. macrocaphala and Poria cocos, respectively, could induce the CYP3A4 gene expression with (6.82 +/- 0.09), (6.76 +/- 0.20), (5.49 +/- 0.13) and (4.97 +/- 0.07) folds, as compared with 0.1% DMSO treated cells. In the time-effect study, 500 mg x L(-1) Rhizoma curcumae, A. lancea, A. macrocaphala and Poria cocos for 48 h could induce the CYP3A4 gene expression with (7.74 +/- 0.54), (7.34 +/- 0.10), (5.54 +/- 0.11) and (5.32 +/- 0.18) folds, compared with 0.1% DMSO treated cells. CONCLUSION: Rhizoma Curcumae, A. lancea, A. macrocaphala and Poria cocos could induce the expression of CYP3A4 gene transcription through activating PXR.