[Establishment of induced pluripotent stem cell lines from human amniotic fluid cells with 1q21.1 microdeletion].

OBJECTIVE: To reprogram the 1q21.1 microdeletion pluripotent stem cells in order to establish an ideal model for further studying its pathogenesis. METHODS: Human amniotic fluid-derived cells induced pluripotent stem cells (hAF-iPSCs) were induced from amniotic fluid cells harboring the 1q21.1 microdeletion by retroviral vectors encoding Oct4, Sox2, c-Myc and Klf4. Characteristics of the 1q21.1 microdeletion hAF-iPSCs were determined, which included in vitro pluripotency, karyotype, microarray analysis, the capacity of differentiation in vivo and in vitro, etc. RESULTS: hAF-iPSCs derived from amniotic fluid cells harboring the 1q21.1 microdeletion have maintained self renewal, with expression of pluripotency marker genes detectable at mRNA level. Stem cell surface antigens were tested by immunocytochemistry. The 1q21.1 microdeletion hAF-iPSCs showed a normal karyotype after long-term culturing in vitro, and harbored the same microdeletion as confirmed by microarray analysis. The cells have maintained their differentiation capacity both in vivo and in vitro. CONCLUSION: The hAF-iPSCs harboring the 1q21.1 microdeletion have all the characteristics of normal pluripotent stem cells, and can be used for directed differentiation into specific cells, which may provide an ideal model for studying the pathogenesis of 1q21.1 microdeletion in vitro.

[Detection for chromosomal aberrations in 43 fetuses with spontaneous abortion and stillbirth by array-based comparative genomic hybridization].

OBJECTIVE: To assess the value of array-based comparative genomic hybridization (array-CGH) for analyzing tissues derived from spontaneous abortion and stillbirth. METHODS: Agilent Human Genome CGH Microarray 4×44 K chip and Affymetrix Cytoscan 750 K Array were utilized to detect genome-wide copy number variations (CNV) in 43 fetuses with spontaneous abortion and stillbirth. All identified CNV were analyzed with references from Database of Genomic variants (DGV), database of DECIPHER, ISCA and OMIM, as well as comprehensive literature review to determine whether the identified CNVs were pathogenic. Parental DNA of two cases was also analyzed with the same arrays for pathogenic or unknown significant CNVs. RESULTS: All of the 43 specimens were successfully analyzed. Clinically significant chromosomal aberrations were identified in 32 (74.4%) of the samples, which included 26 aneuploidies and 10 pathogenic CNV. CONCLUSION: Array-CGH is a fast and effective method for analyzing tissues derived from spontaneous abortions and stillbirths which may be difficult to culture for karyotype analysis.

Have Previous COVID-19 Vaccinations Shaped the Potential Enhancing Infection of Variant Strains?

OBJECTIVE: This study aimed to investigate the infection status of Omicron in the population and the association between COVID-19 vaccination and infection with Omicron. METHODS: We conducted a cross-sectional study to openly recruit participants for a survey of SARS-CoV-2 infection by convenience sampling from 1 January to 15 January 2023 after a COVID-19 pandemic swept across China. Additionally, the binary logistic regression model was adopted to evaluate the association between COVID-19 vaccination and the infection outcomes or symptom severity, respectively. Meanwhile, the relations between the vaccination and duration of the symptoms were estimated via ordinal logistic analysis. RESULTS: Of the 2007 participants, the prevalence of infection with Omicron was 82.9%. Compared with unvaccinated individuals, inactivated COVID-19 vaccination could increase the risk of Omicron infection (OR = 1.942, 95% CI: 1.093-3.448), and the receipt of at least one dose of non-inactivated COVID-19 vaccines was a protective factor against infection (OR = 0.428, 95% CI: 0.226-0.812). By contrast, no relations were observed in COVID-19 vaccination with the symptoms of infection and duration of symptoms (p > 0.05). CONCLUSIONS: This cross-sectional study concluded that inactivated COVID-19 vaccination might increase the risk of Omicron infection, which should be a concern during COVID-19 vaccination and the treatment of variant infections in the future, and the receipt of at least one dose of non-inactivated COVID-19 vaccine was a protective factor against infection.

Supramolecular Hydrogel Dexamethasone-Diclofenac for the Treatment of Rheumatoid Arthritis.

Rheumatoid arthritis (RA) severely affects patients' quality of life and is commonly treated with glucocorticosteroids injections, like dexamethasone, which may have side effects. This study aimed to create a novel low dose of twin-drug hydrogel containing dexamethasone and diclofenac and explore its potential as a drug delivery system for an enhanced anti-inflammatory effect. Its characterization involved rheology, transmission electron microscope (TEM), Fourier-transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD). Furthermore, the hydrogel demonstrated thixotropic properties. The hydrogel exhibited no cytotoxicity against RAW 264.7 macrophages. Furthermore, the hydrogel demonstrated a significant anti-inflammatory efficacy by effectively downregulating the levels of NO, TNF-α, and IL-6 in lipopolysaccharide (LPS)-activated RAW 264.7 macrophages. The co-delivery approach, when intra-articularly injected in adjuvant-induced arthritis (AIA) rats, significantly alleviated chronic inflammation leading to reduced synovitis, delayed bone erosion onset, and the downregulation of inflammatory cytokines. The biocompatibility and adverse effect evaluation indicated good biological safety. Furthermore, the hydrogel demonstrated efficacy in reducing NF-κB nuclear translocation in LPS-induced RAW 264.7 macrophages and inhibited p-NF-kB, COX-2, and iNOS expression both in RAW 264.7 macrophages and the joints of AIA rats. In conclusion, the findings indicate that the hydrogel possesses potent anti-inflammatory activity, which effectively addresses the limitations associated with free forms. It presents a promising therapeutic strategy for the management of RA.

Chromatographic performance of zidovudine imprinted polymers coated silica stationary phases.

Nowadays, molecularly imprinted polymers (MIPs) coated silica stationary phases (SPs) have aroused great attention, owing to their good properties of high selectivity, good stability, facile synthesis procedure and low cost. In this study, zidovudine imprinted polymers coated silica stationary phases (MIPs/SiO2 SPs) were synthesized by surface imprinting technique using zidovudine as the template molecule, methacrylic acid as the functional monomer, ethylene glycol dimethacrylate as the cross-linking agent, azobisisobutyronitrile as the initiator, and bare silica spheres (particle size, 5 µm; pore size, 20 nm) as substrates. In the process, reagents with low concentration were used to prepare thin layer of MIPs coating on the surface of silica microbeads. The properties of the materials were characterized by scanning electron microscope (SEM), fourier transform infrared spectrometer (FT-IR), carbon elemental analysis and N2 adsorption-desorption experiment. The obtained SPs were packed into stainless steel columns (2.1 mm × 150 mm) via a slurry method. The prepared columns were applied for separation of nucleoside analogues with similar chemical structures and strong polarity. The retention mechanism of MIPs/SiO2 SPs for nucleoside analogues was investigated carefully. And the chromatographic performances of the resulting MIPs based SPs were superior to those of the commercial SPs. Furthermore, the synthesized MIPs/SiO2 SPs possessed great potentials in separation of ginsenosides. This investigation demonstrated that MIPs based SPs were successfully synthesized and provided a new approach to polar compounds separation and analysis.

Follicular Fluid-Derived Exosomal MicroRNA-18b-5p Regulates PTEN-Mediated PI3K/Akt/mTOR Signaling Pathway to Inhibit Polycystic Ovary Syndrome Development.

Small RNA sequences in follicular fluid (FF)-derived exosomes (extracellular vesicles contain proteins, DNA, and RNA) vitally function in the development of polycystic ovary syndrome (PCOS). It has been identified that microRNA (miR)-18b-5p is one of miRs that differ between control and PCOS women that passed the false discovery rate, and phosphatase and tensin homolog deleted on chromosome 10 (PTEN) is an important modifier of biological functions of ovarian granulosa cells (GCs) in PCOS. However, whether miR-18b-5p could functionally mediate the progression of PCOS via PTEN was not clarified completely, which was the issue we wanted to solve in our research. FF-derived exosomes were isolated using an extraction kit. KGN cells were co-cultured with miR-18b-5p-modified exosomes or transfected with a PTEN-related vector. After treatment, cell proliferation and apoptosis were observed. A rat model of PCOS was established by letrozole and then injected with miR-18b-5p-modified exosomes. Then, serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone, and estradiol (E2) levels in PCOS rats were measured. miR-18b-5p, PTEN, and phosphatidylinositol 3 kinases/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) pathway-related genes were tested. In PCOS patients, miR-18b-5p was downregulated, and PTEN was highly expressed in FF and GCs. PTEN knockdown increased KGN cell proliferation and limited apoptosis. FF-derived exosomes stimulated proliferation and suppressed apoptosis of KGN cells; decreased FSH, LH, and testosterone; and increased E2 in PCOS rats. Upregulating miR-18b-5p further enhanced the inhibitory effects of exosomes on suppressing the progression of PCOS. miR-18b-5p targeted PTEN and could activate PI3K/Akt/mTOR pathway. miR-18b-5p produced by FF-derived exosomes reduces PTEN expression and promotes the activation of the PI3K/Akt/mTOR signaling pathway to improve PCOS. Based on that, circulating miR-18b-5p levels can contribute to the progression of PCOS complications.

ROS-mediated liposomal dexamethasone: a new FA-targeted nanoformulation to combat rheumatoid arthritis via inhibiting iRhom2/TNF-α/BAFF pathways.

Rheumatoid arthritis (RA) is an autoimmune inflammatory disorder that has seriously affected human health worldwide and its current management requires more successful therapeutic approaches. The combination of nanomedicines and pathophysiology into one system may provide an alternative strategy for precise RA treatment. In this work, a practical ROS-mediated liposome, abbreviated as Dex@FA-ROS-Lips that comprised synthetic dimeric thioether lipids (di-S-PC) and a surface functionalized with folic acid (FA), was proposed for dexamethasone (Dex) delivery. Incorporation with thioether lipids and a FA segment significantly improved the triggered release and improved the triggered release of cytotoxic Dex as well as the active targeting of RA, altering its overall pharmacokinetics and safety profiles in vivo. As proof, the designed Dex@FA-ROS-Lips demonstrated effective internalization by LPS-activated Raw264.7 macrophages with FA receptor overexpression and released Dex at the inflammatory site due to the ROS-triggered disassembly. Intravenous injection of this Dex@FA-ROS-Lips into adjuvant-induced arthritis (AIA) mice led to its incremental accumulation in inflamed joint tissues and significantly alleviated the cartilage destruction and joint swelling via suppression of proinflammatory cytokines (iRhom2, TNF-α and BAFF), as compared to the effect of commercial free Dex. Importantly, the Dex@FA-ROS-Lips nanoformulation showed better hemocompatibility with less adverse effects on the body weight and immune organ index of AIA mice. The anti-inflammatory mechanism of Dex@FA-ROS-Lips was further studied and it was found that it is possibly associated with the down-regulation of iRhom2 and the activation of the TNF-α/BAFF signaling pathway. Therefore, the integration of nanomedicines and the RA microenvironment using multifunctional Dex@FA-ROS-Lips shall be a novel RA treatment modality with full clinical potential, and based on the enhanced therapeutic effect, the signaling pathway of iRhom2/TNF-α/BAFF reasonably explained the mechanism of Dex@FA-ROS-Lips in anti-RA, which suggested a molecular target for RA therapy and other inflammatory diseases.

Inter-individual variations and modulators of MDR1 transport activity in human placenta.

INTRODUCTION: ATP binding cassette (ABC) membrane transporter multidrug resistance 1 (MDR1) is one of the most important efflux transporters in the human placenta protecting the fetus from exposure to xenobiotic toxicity. Recent studies have focused on placental MDR1 expression, but few studies have analyzed placental MDR1 transport activity. The purpose of this study was to investigate placental MDR1 transport activity using a relatively large sample size of human placentas. Furthermore, the effect of ABCB1 gene polymorphisms were investigated along with physiological factors including maternal age, times of pregnancy, BMI, delivery mode or pregnancy complications on placental MDR1 transport activity. METHODS: A total of 252 human placentas were obtained after delivery. MDR1 transport activity was detected by N-methyl quinidine uptake in placental microvillus membrane vesicles (MVMVs). Nine common single nucleotide polymorphisms (SNPs) in ABCB1 genes were determined by Snapshot. The association between ABCB1 gene polymorphisms, maternal age, times of pregnancy, BMI, delivery mode or pregnancy complications, and transporter activity was investigated. RESULTS: Inter-individual variations of MDR1 transport activity were observed among 252 subjects. The per unit protein activity was ranged from 0.05 to 0.15/mg. Nine SNPs in ABCB1 gene didn't exhibit significant association with transporter activity of MDR1. Likewise, neither age, times of pregnancy, delivery mode nor pregnancy complications showed any significant effect of placental MDR1 transport activity. But placental MDR1 transport activity in obese pregnant women was lower than those in non-obese pregnant women. CONCLUSION: Inter-individual variations of MDR1 transport activity existed in human placentas. This may contribute to variations in drug exposure to the fetus affecting clinical outcomes. Maternal age, times of pregnancy, delivery mode nor pregnancy complications included in this study maybe not significantly impact placental MDR1 transport activity, but maternal obese could inhibit placental MDR1 activity.

Effect of proton pump inhibitors on high-dose methotrexate elimination: a systematic review and meta-analysis.

Background Drug interaction is one factor which may influence high-dose methotrexate (MTX) elimination. Proton pump inhibitors are commonly used as an adjuvant drugs in chemotherapy. However, the effect of proton pump inhibitors on high-dose MTX elimination is currently controversial. Objective To perform a systematic review and meta-analysis to assess the association between co-administration of proton pump inhibitors with plasma MTX concentration and delayed MTX elimination. Setting The Hospital of Kunming Medical University, China. Method We followed the PRISMA guidelines in this meta-analysis and systemic review. We searched PubMed, the Cochrane Database, Embase, the WHO International Clinical Trials Registry Platform, the Wanfang database, the Chinese National Knowledge Infrastructure, the VIP database and the Chinese BioMedical Literature Database. Main outcome measure The main outcome measures are: (1) the plasma MTX concentration at 24 h, 48 h and 72 h.; (2) the frequency of patients with delayed MTX elimination. Results Ten retrospective cohort studies were included in the meta-analysis, with a total of 2760 cycles of high-dose MTX treatment. A meta-analysis revealed that compared to patients who did not receive proton pump inhibitors, patients who received proton pump inhibitors had a significantly higher plasma MTX concentration at 24 h (mean difference 2.71 µM, 95% confidence interval 0.55 to 4.87; p = 0.01) and at 48 h (mean difference 0.14 µM, 95% confidence interval 0.06 to 0.21; p < 0.01) after the MTX infusion. Furthermore, delayed MTX elimination was more frequent in patients that received PPIs (risk ratio 0.59, 95% confidence interval 0.41 to 0.84; p = 0.004). Conclusion This systematic review and meta-analysis reveals that the co-administration of proton pump inhibitors with methotrexate is associated with delayed high-dose MTX elimination. Proton pump inhibitors should be cautiously given when co-administered with high-dose MTX treatment.

[Multicentric randomized double blinded clinical study of Yiqi Tongmai Oral Liquid against angina pectoris in patients with coronary heart disease].

OBJECTIVE: To study the efficacy and safety of Yiqi Tongmai Oral Liquid (YQTM), a traditional compound Chinese herbal medicine, in treating angina pectoris in patients with coronary heart disease. METHODS: A multicentric, randomized, double blinded and paralleled controlled trial was conducted on 110 patients in trial group treated with YQTM, and 109 patients in control group treated with Shuxin Oral Liquid (SX). Cure and effective rates in both groups were evaluated. Frequency and duration of angina attack were counted and measured. Coronary angiography (CAG), electrocardiogram (ECG) and flat exercise test were taken in both groups. Blood lipid indexes, such as cholesterol (CH), triglyceride (TG), low-density lipoprotein (LDL), high density lipoprotein (HDL), were determined at pre- and post-treatment. The hemodynamic indexes, such as whole blood viscosity (J2), high-shear reduced viscosity (Eh), low-shear reduced viscosity (Ei), red cell aggregation index (Lb), red cell rigidity index (Rh), fibrinogen (Fb), blood sedimentation rate (BSR) and hematocrit (HCT), were determined at pre-and post-treatment. The indicated scores of symptoms and signs of traditional Chinese medicine (TCM) pattern, such as chest pain, chest constriction, breath shortness, palpitation, fatigue, dim complexion, spontaneous perspiration and tongue proper, tongue coating were evaluated in week 0, 1, 2, 3, 4 during the treatment course. The safety indexes, such as body temperature, pulse, respiration and blood pressure were observed. Routine tests of blood, urine and stool, hepatic function test and renal function test were taken at pre- and post-treatment. RESULTS: There was no significant difference between the total effective rate of the trial group and that of the control group, which were 91.82% and 85.32%, respectively (P>0.05). Trial groups percentile of cure rate is significantly higher than that of the control group (P<0.01). The frequency and duration of angina attack, the positive ratio of CAG and flat exercise test of both groups were lowered, while the effect of the trial group on frequency and duration of angina attack was better. No significant difference was found in ECG features between the two groups (P>0.05). The levels of CH, TG and LDL of both groups were lowered significantly (P<0.05). The effect of lowering CH, TG and LDL of the trial group was stronger than that of the control group (P<0.05). The hemodynamic indexes, such as J2, Eh, Ei, Lb, Rh, Fb, BSR and HCT were improved significantly in both groups (P<0.05). The improvements of J2, Eh, Ei, Lb, Rh, Fb and SR in the trial group were greater than those of control group (P<0.05). The TCM symptoms and signs, such as chest pain, chest constriction, breath shortness, palpitation, fatigue, dim complexion, spontaneous perspiration were improved significantly in both groups (P<0.05). The improvements of chest constriction, palpitation, fatigue and spontaneous perspiration in the trial group were greater than those of the control group (P<0.05). The total indicated score of TCM symptoms and signs was lowered more significantly than that of the control group (P<0.01). No significant changes were found at pre- and post-treatment in safety indexes, such as routine tests for blood, urine and stool, hepatic function test and renal function test. There was no significant difference in safety features of both groups (P>0.05). CONCLUSION: Yiqi Tongmai Oral Liquid bears good therapeutic effect on angina pectoris without adverse reaction, and is superior to Shuxin Oral Liquid. Yiqi Tongmai Oral Liquid is a new effective and safe medicine for the treatment of angina pectoris in patients with coronary heart disease.

Combined treatment with low dose prednisone and escin improves the anti-arthritic effect in experimental arthritis.

The present study was aimed at investigating whether low dose oral prednisone combined with escin could inhibit the progression of adjuvant-induced arthritis (AIA) in rats. Adjuvant arthritis was induced in SD rats began day 1 for 28 days. Prednisone at doses of 2, 10 mg/kg/day alone or escin at doses of 5, 10 mg/kg/day alone, or prednisone at dose of 2 mg/kg/day with escin at doses of 5 or 10 mg/kg/day were given to different groups of rats intragastrically from day 13 to 28 respectively. Paw swelling, arthritic index, histological and radiographic changes were assessed to evaluate the anti-arthritic effect. Weight growth, spleen and thymus indexes were also calculated. Serum samples were collected for estimation of pro-inflammatory cytokines. Rats developed erosive arthritis of the hind paw when immunized with adjuvant. Prednisone 2 mg/kg combined with escin 5 or 10 mg/kg significantly inhibited the paw swelling. Histopathological and radiographic analysis showed a marked decrease of synovial inflammatory infiltration, synovial hyperplasia and bone erosion by combination therapy, which also markedly suppressed the expression of tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß) and interleukin-6 (IL-6). No significant changes were found in monotherapy group except prednisone 10 mg/kg group. Furthermore, combined treatment rescued some of GCs' adverse effects evidenced by increase in body weight and decrease in index of spleen compared with untreated AIA rats. In conclusion, the combination therapy possessed synergistic anti-arthritic efficacy and reduced adverse effect, which may play a role in the management of human RA.

Prenatal diagnosis of hemivertebrae--A likely association with 7q deletion.

OBJECTIVE: This study aims to investigate the possible cause of a prenatal case of hemivertebrae with a 7q terminal deletion. CASE REPORT: This case describes a fetus with hemivertebrae in thoracic vertebrae as the sole antenatal sonographic finding. Genetic testing was performed in order to find more information after the abnormal ultrasound finding. The array-based comparative genomic hybridization results showed that the fetus had approximately 6.4 Mb deletion of 7q36. We discussed the two genes (SHH and HLXB9) that may be associated with hemivertebrae in the deletion region and reviewed several literatures about 7q36 deletion. CONCLUSION: Our results suggest that the phenotype of hemivertebra in our case may be related to the deletion of 7q36.

Yanshu spraying agent, a traditional Chinese medicine, relieves chronic pharyngitis in animals by anti-inflammatory and antibacterial effects.

Chronic pharyngitis is chronic inflammation that is often caused by repeated occurrences of acute pharyngitis or upper respiratory tract infections, including Streptococcus and Staphylococcus. The present study aimed to investigate the effects of Yanshu spraying agent (Yanshu) in relieving chronic pharyngitis, as well as the possible underlying mechanisms. The results revealed that Yanshu inhibited chronic inflammation in ammonia-induced chronic pharyngitis in rabbits and cotton pellet-induced granuloma tissue formation in rats. Yanshu also demonstrated antibacterial effects on Streptococcus and Staphylococcus in vitro. Yanshu did not exhibit any effects on the immune system, including the spleen and thymus indexes, immunocyte count and monocyte-macrophage function, when compared with the effects of dexamethasone. Therefore, the results of the present study indicate that Yanshu may relieve chronic pharyngitis via its anti-inflammatory and antibacterial activities.

The effects of huperzine A on gastrointestinal acetylcholinesterase activity and motility after single and multiple dosing in mice.

The acetylcholinesterase inhibitor (AChEI), huperzine A has been used in the treatment of the cognitive deterioration associated with Alzheimer's disease (AD). However, the side-effects of huperzine A associated with increased cholinergic activity, particularly in the gastrointestinal system, are evident. It is not yet known how quickly these side-effects become tolerated; this information would provide guidance to doctors on how to use huperzine A so as to attenuate the adverse events. The present study aimed to observe the effects of huperzine A on gastrointestinal motility and acetylcholinesterase (AChE) activity in mice. After oral administration of huperzine A with single and multiple dosing, the gastrointestinal motility and AChE activity of the mice were examined. The results revealed that, following a single dose of huperzine A, the AChE activity in the stomach and duodenum were significantly inhibited and the gastrointestinal motility was significantly increased. However, following multiple doses (7 or 28 doses, one dose per day), no significant changes in the AChE activity and gastrointestinal motility were identified. These findings indicate that the gastrointestinal adverse effects of huperzine A may be well-tolerated relatively quickly and do not recur. Additionally, it suggests that patients with AD are likely to have minimal gastrointestinal side-effects after taking multiple doses of huperzine A.

Ginsenoside Rg1 exerts synergistic anti-inflammatory effects with low doses of glucocorticoids in vitro.

Glucocorticoids (GCs) are usually used to treat inflammatory diseases. However, they cause severe and irreversible side effects, which limit the use of these compounds. Ginsenoside Rg1 had been demonstrated to possess anti-inflammatory and anti-cancer effects. The present study was designed to investigate whether Rg1 exhibits synergistic anti-inflammatory effects when combined with glucocorticoids. After stimulated by lipopolysaccharide (LPS), murine macrophagic RAW264.7 cells were treated with Rg1, corticosterone (Cort) or Rg1 and Cort. Then nitric oxide (NO), tumor necrosis factor-α (TNF-α) and glucocorticoid receptor (GR) expression were measured. The results showed that Rg1 or Cort could reduce the production of NO and TNF-α, and Rg1 dose-dependently up-regulated GR expression, while Cort dose-dependently down-regulated GR expression. The combination of low concentrations of Rg1 with Cort, which alone could not markedly inhibit the release of inflammatory factors, inhibited the secretion of NO and TNF-α in LPS-stimulated RAW264.7 macrophage cells, and up-regulated the expression of GR. The findings suggested Rg1 can synergize with glucocorticoid to enhance its anti-inflammatory effect.