UCH-L1 Inhibitor Alleviates Nerve Damage Caused by Moyamoya Disease.

Background: Moyamoya disease (MMD) leads to nerve injury. Exosomes are touted as bio-shuttles for the delivery of distinct biomolecules inside the cells. Recently, UCH-L1 was shown to play a vital role in nerve injury. However, it is still unknown whether UCH-L1 can improve the nerve injury of MMD. Materials and Methods: Exosomes were isolated from the serum of patients with MMD and healthy controls. The total RNA was extracted from the exosomes, and the level of GFAP and UCH-L1 between the serum exosomes of the two groups was analyzed by a quantitative reverse transcription-polymerase chain reaction and western blot. Exosome labeling and uptake by SH-SY5Y cells were observed by confocal laser microscopy. Cell counting kit-8 assay and flow cytometry were used to determine the viability and apoptosis of SH-SY5Y cells, respectively. Results: Exosomes were successfully isolated and identified from serum. The expression of GFAP and UCH-L1 was significantly higher in the serum-derived exosomes from MMD patients compared with the healthy controls (P < 0.05). Compared to the blank and control exosome group, serum-derived exosomes from MMD significantly suppress cellular vitality and promote apoptosis of SH-SY5Y cells, while the use of LDN-91946, a specific inhibitor of UCH-L1, could reverse the effects induced by serum-derived exosomes from MMD. Conclusion: UCH-L1 inhibitor could reverse MMD-induced inhibition of SH-SY5Y cell viability and promotion of apoptosis. UCH-L1 may be a therapeutic target for the treatment of nerve damage caused by MMD.

Expert consensus on multidisciplinary approach to the diagnosis and treatment of primary hyperparathyroidism in pregnancy in China.

Primary hyperparathyroidism in pregnancy is a rare disease that can have detrimental effects on both maternal and fetal/neonatal outcomes. The physiological changes that occur during pregnancy can complicate the diagnosis, imaging examinations, and treatment of this disorder. To enhance our understanding and management of primary hyperparathyroidism in pregnancy, experts from various fields, including endocrinology, obstetrics, surgery, ultrasonography, nuclear medicine, pediatrics, nephrology, and general practice in China, collaborated to develop a consensus addressing the critical aspects of the diagnosis and treatment of primary hyperparathyroidism in pregnancy with a multidisciplinary team approach. This consensus provides valuable guidance for healthcare professionals in managing this condition, ultimately improving outcomes for both mothers and their babies.

Multidisciplinary team efforts to improve the pregnancy outcome of pregnancy complicated with primary hyperparathyroidism: case series from a single hospital.

BACKGROUND: There is no consensus or management algorithm for primary hyperparathyroidism (PHPT) in pregnancy. METHODS: This study comprises a retrospective case series. From August 2014 to December 2020, 9 cases of PHPT in pregnancy were diagnosed by a multidisciplinary team (MDT) consultation center of obstetrics in our hospital. Their clinical manifestations, treatment strategies, and maternal and infant outcomes were analyzed. RESULTS: The median onset age of the patients was 32 (25 ~ 38) years. PHPT was diagnosed in two cases before pregnancy, in six cases during pregnancy and in one case postpartum. The main clinical manifestations were nausea, vomiting, and other nonspecific symptoms, with anemia as the most common maternal complication. Hypercalcemia crisis was developed in one case. The median levels of preoperative serum calcium and parathyroid hormone (PTH) were 3.08 (2.77 ~ 4.21) mmol/L and 300.40 (108.80 ~ 2603.60) pg/ml, respectively. The parathyroid ultrasonography tests were positive in eight cases and negative in one patient who had an ectopic lesion localized by 99mTc-MIBI. Parathyroidectomy was conducted in 7 cases during the 2nd trimester, including 2 patients diagnosed before pregnancy who refused surgery, 1 patient during the 1st trimester, and 1 patient postpartum, with a significant reduction in serum concentrations of calcium and PTH. A management algorithm was developed. CONCLUSION: This case series suggests that pregnant women with PHPT should be managed by MDT according to the algorithm. If PHPT is confirmed in fertile women before pregnancy, parathyroidectomy should be strongly suggested and performed. If PHPT is diagnosed during pregnancy, even in its mild form, surgical treatment, optimally during the 2nd trimester, is effective and safe for pregnancy and neonatal outcome.

Diagnosis, fetal risk and treatment of pemphigoid gestationis in pregnancy: A case report.

BACKGROUND: Pemphigoid gestationis (PG) is a rare autoimmune blistering disease that usually presents in the second or third trimester, with an incidence of 1 per 50000 pregnancies. PG tends to recur with an earlier onset and a more severe course in subsequent pregnancies. Skin biopsy markers can be confirmed by direct immunofluorescence staining. CASE SUMMARY: Our patient was diagnosed with PG at 8 mo of gestation with fresh bullous lesion marks on the abdomen and limbs. Termination of the pregnancy was performed by cesarean section at 37 + 4 wk of gestation. The patient delivered an infant weighing 3620 gm. The infant had urticaria-like and vesicular skin lesions and was diagnosed with PG. The patient was discharged on prednisolone and in a satisfactory condition. The infant was discharged after anti-inflammatory therapy for one week. CONCLUSION: PG is a rarely reported disease, and 10% of newborns develop mild clinical symptoms consisting of urticaria-like or vesicular skin lesions. We intend to remind clinicians to consider this condition when a patient presents with such lesions so that treatment can be started early and neonatal morbidity can be taken into account.

The Sex Specific Association Between Maternal Gestational Diabetes and Offspring Metabolic Status at 1 Year of Age.

Previous studies showed the association between maternal GDM and long-term effects of overweight in offspring. However, the nature of this association in the early postnatal period is still undetermined. The aim of this prospective cohort study was to evaluate whether maternal GDM is associated with overweight and obesity status in offspring at age 1 year. We studied 1167 infants born at a large obstetrical care hospital including 778 normal glucose tolerance (NGT) and 389 GDM pregnancies, matched in a 1:2 ratio according to offspring's gender, during the years 2016-2017. Overweight and obesity status in offspring of both groups were evaluated at 1 year of age through questionnaires. Infant outcomes were defined according to the WHO Child Growth Standards based on the length-based BMI-for-age. Female offspring from the GDM group exhibited a higher mean BMI (17.2 vs. 16.6, p < 0.01), a higher rate of obesity (13.9% vs. 7.7%; p < 0.05), and overweight (33.1% vs. 23.5%; p < 0.05) as compared to the NGT female group. In the multivariable regression model, maternal GDM was found to be independently and significantly associated with overweight or obesity in 1-year aged female offspring only (OR 1.61, 95% CI 1.09-2.37, p < 0.05). We found a sex specific association between maternal GDM and the overweight risk only in female offspring at 1 year of age.

Label-Free Fluorescence-Based Aptasensor for the Detection of Sulfadimethoxine in Water and Fish.

Fluorescence-based aptasensors possess high sensitivity but are complicated and usually require multistep labeling and modification in method design, which severely limit the practical applications. Here, a label-free fluorescence-based aptasensor, consisting of aptamer, gold nanoparticles (AuNPs), and cadmium telluride (CdTe) quantum dots (QDs), was developed for the detection of sulfadimethoxine (SDM) in water and fish based on the specific recognition of SDM-aptamer and the inner filter effect of QDs and AuNPs. In the absence of a target, AuNPs dispersed in salt solution because of the aptamer protection, which could effectively quench the fluorescence emission of QDs, while in the presence of SDM, AuNPs aggregated due to the specific recognition of SDM-aptamer to SDM, which resulted in fluorescence recovery. A linear response of SDM concentrations in the range of 10-250 ng mL-1 ( R2 = 0.99) was obtained, and the detection limit was 1.54 ng mL-1 (3σ, n = 9), far below the maximum residue limit (100 ng mL-1) of SDM in edible animal tissues regulated by China and the European Commission. The fluorescence-based aptasensor was applied to the detection of SDM in aquaculture water and fish samples with high accuracy, excellent precision, and ideal selectivity. The results indicated that the developed aptasensor was simple in design, easy to operate, and could be used to detect rapidly and accurately SDM in water and fish samples.

A highly selective and sensitive colorimetric detection of uric acid in human serum based on MoS2-catalyzed oxidation TMB.

A simple, rapid, and effective colorimetric assay for the detection of uric acid (UA) has been built, using the MoS2 nanoflakes-catalyzed 3,3',5,5'-tetramethylbenzidine (TMB)-H2O2 system. In the presence of oxygen and uricase, uric acid was oxidized specifically to produce H2O2. MoS2 nanoflakes synthesized by hydrothermal reaction could catalyze the oxidation of TMB by H2O2, and engendering the colorimetric signal. Finally, the change in the color from colorless to blue was seen with naked eye, indicating the presence or absence of UA. Under the optimized conditions, a linear relationship between the absorbance and UA concentration in the range of 0.5-100 µM (R2 = 0.996) with the limit of detection for 0.3 µM (S/N = 3). The proposed assay was successfully applied to the detection of UA in human serum with the recoveries over 94.54%. Thus, these results suggest that the UA assay-based MoS2-catalyzed TMB-H2O2 has great foreground for fast clinical diagnosis of gout without the need for advanced and costly equipment. Graphical Abstract ᅟ.

Detection of malachite green in fish based on magnetic fluorescent probe of CdTe QDs/nano-Fe3O4@MIPs.

A magnetic fluorescent probe of CdTe QDs/nano-Fe3O4@MIPs was prepared using CdTe QDs and Fe3O4 nanoparticles as co-nucleus and molecularly imprinted polymers (MIPs) as specific recognition sites based on a reverse microemulsion method. With the specific enrichment and magnetic separation properties, the probe of CdTe QDs/nano-Fe3O4@MIPs was used to detect malachite green (MG) in fish samples. The TEM analysis showed that the particles of CdTe QDs/nano-Fe3O4@MIPs were spherical with average diameter around 53nm, and a core-shell structure was well-shaped with several Fe3O4 nanoparticles and CdTe QDs embedded in each of the microsphere. Quick separation of the probes from solutions could be realized with a magnet, indicating the excellent magnetic property of CdTe QDs/nano-Fe3O4@MIPs. The probe exhibited high specific adsorption towards MG and excellent fluorescence emission at λem 598nm. The fluorescence of CdTe QDs/nano-Fe3O4@MIPs could be linearly quenched by MG at the concentrations from 0.025 to 1.5µmolL-1. The detection limit was 0.014µmolL-1. The average recovery of spiked MG in fish samples was 105.2%. The result demonstrated that the as-prepared CdTe QDs/nano-Fe3O4@MIPs could be used as a probe to the detection of trace MG in fish samples.

Biomimetic ELISA detection of malachite green based on molecularly imprinted polymer film.

A highly selective and sensitive enzyme-linked immunosorbent assay (ELISA) was developed for the detection of malachite green (MG) using a molecularly imprinted polymer (MIP) film as bionic antibody. The MIP film, based on the self-polymerization of dopamine, was fabricated on the surfaces of a 96-well microplate. It showed specific recognition for MG in aqueous solution. A direct competitive ELISA method was established with the sensitivity reaching 10.31µgL-1 and the detection limit being 0.3µgL-1. The cross-reactivity of two structural analogues to MG was less than 10%. The average recovery tested by MG standard spiking was 88.8% for bass and 90.4% for water, and the relative standard deviations were less than 3.6%. All the above results indicated that the developed method could be used to detect MG in fish and water samples rapidly, specifically and accurately.

Rapid detection of malachite green in fish based on CdTe quantum dots coated with molecularly imprinted silica.

A sensitive fluorescence sensor for the detection of malachite green (MG) was fabricated by grafting molecularly imprinted polymers (MIPs) onto the surface of CdTe quantum dots (QDs). The MIP-coated QDs were synthesized via a reverse microemulsion method using (3-aminopropyl)triethoxysilane (APTES) and tetraethyl orthosilicate (TEOS) as functional monomer and cross-linker, respectively. The optimum molar ratio of MG, functional monomer and cross-linker was 1:3:10. The MIP-coated QDs exhibited uniform spheres with diameter around 49nm and excellent fluorescence emission at λex 370nm. A linear relationship with two segments between the relative fluorescence intensities and the MG concentrations ranging from 0.08 to 20µmol·L-1 could be obtained with a detection limit of 12µg·kg-1. The fluorescent probe was successfully applied to the determination of MG in fish samples with the spiked recoveries ranging from 94.3% to 109.5% which were in accordance with those of the measurement by HPLC-UV.

Biomimetic ELISA detection of malachite green based on magnetic molecularly imprinted polymers.

A direct competitive enzyme-linked immunosorbent assay (ELISA) method was used for the detection of malachite green (MG) with a high sensitivity and selectivity using magnetic molecularly imprinted polymers (MMIPs) as a bionic antibody. MMIPs were prepared through emulsion polymerization using Fe3O4 nanoparticles as magnetic nuclei, MG as a template, methacrylic acid (MAA) as a functional monomer, ethylene glycol dimethacrylate (EGDMA) as a crosslinking agent and span-80/tween-80 as mixed emulsifiers. The MMIPs were characterized by scanning electron micrographs (SEM), thermal-gravimetric analyzer (TGA), Fourier transform infrared spectrometer (FT-IR) and vibrating sample magnetometer (VSM), respectively. A high magnetic saturation value of 54.1emug-1 was obtained, resulting in rapid magnetic separation of MMIPs with an external magnet. The IC50 of the established ELISA method was 20.1µgL-1 and the detection limit (based on IC85) was 0.1µgL-1. The MMIPs exhibited high selective binding capacity for MG with cross-reactivities less than 3.9% for MG structural analogues. The MG spiking recoveries were 85.0%-106% with the relative standard deviations less than 4.7%. The results showed that the biomimetic ELISA method by using MMIPs as bionic antibody could be used to detect MG rapidly in fish samples with a high sensitivity and accuracy.

[Effect of different surgical methods on leprosy plantar ulcers].

OBJECTIVE: To explore the effects of different surgical methods on plantar ulcers in leprosy. METHODS: The clinical data of 71 patients with leprosy plantar ulcers and treated with different surgical methods between October 1950 and October 2006 were analyzed retrospectively. In group A, 34 cases underwent debridement, including 26 males and 8 females aged 53-88 years old (average 72.4 years old); the course of ulcer averaged 29.0 years; the size of ulcer ranged from 5 cm x 3 cm x 2 cm to 11 cm x 7 cm x 3 cm; the disability degrees of the affected foot was mild in 25 cases and severe in 9 cases according to the self-designed evaluation system. In group B, 22 cases received foot pressure rebuilding surgery, including 19 males and 3 females aged 48-83 years old (average 69.8 years old); the course of ulcer averaged 33.5 years; the size of ulcer ranged from 5 cm x 3 cm x 2 cm to 12 cm x 7 cm x 3 cm; the disability degrees of the affected foot was mild in 12 cases and severe in 10 cases. In group C, 15 cases were repaired with the transposition of toe flap, foot arch flap, acrotarsium flap, or medial tibia flap, including 11 males and 4 females aged 43-73 years old (average 64.6 years old); the course of ulcer averaged 29.3 years; the size of ulcer ranged from 6 cm x 3 cm x 2 cm to 11 cm x 5 cm x 3 cm; the disability degrees of the affected foot was mild in 9 cases and severe in 6 cases. No significant differences were evident among three groups in terms of the general information (P > 0.05), except for the difference between group A and group C on age (P < 0.05). RESULTS: Group A: 19 out of 34 cases healed and the average healing time was 46.8 days; all patients were followed up for 2-45 years (average 17.2 years); the rate of ulcer healing 1 year after operation was 55.9% (19/34); 12 healed ulcer patients relapsed at average 1.5 years after operation; the rate of ulcer healing at last follow-up was 20.6% (7/34). Group B: 18 out of 22 cases healed and the average healing time was 29.2 days; all patients were followed up for 2-50 years (average 13.3 years); the rate of ulcer healing 1 year after operation was 81.8% (18/22); 7 healed ulcer patients relapsed at average 3.3 years after operation; the rate of ulcer healing at last follow-up was 50.0% (11/22). Group C: 14 out of 15 cases healed and the average healing time was 27.1 days; all patients were followed up for 3-12 years (average 8.8 years). The rate of ulcer healing 1 year after operation was 93.3% (14/15); 7 healed ulcer patients relapsed at average 4 years after operation; the rate of ulcer healing at final follow-up was 46.7% (7/15). For the rate of ulcer healing 1 year after operation, there was a significant difference between group A and group B, and between group A and group C (P < 0.05), but no significant difference was evident between group B and group C (P > 0.05). For the rate of ulcer healing at the final follow-up visit, there was a significant difference between group A and group B (P < 0.05), but no significant difference was evident between group A and group C, and between group B and group C (P > 0.05). CONCLUSION: The surgical treatment of plantar ulcers in leprosy should include the alleviation of the plantar high-pressure zone and the transposition of the flaps, providing good short-term and long-term therapeutic effect.

Screening of high cytotoxic tumor killer cells using a sensitive adherent target detachment assay.

Screening of high cytotoxic tumor killer cells is of great importance in adoptive immunotherapy. Here, we describe a more sensitive assay, as compared to traditional 51Cr- or Calcein-release assay, to measure cytolytic activity of killer cells. This adherent target detachment (ATD) assay is carried out in microwells of Terasaki tissue culture trays using adherent tumor cells as targets. Target tumor cells are seeded at the concentration of 300-400 cells/well and incubated overnight to allow for the adhesion of cells to the plastic surface of the wells. Effector cells were added at various effector: target (E:T) ratios, and incubated for 24 h. During incubation, dead target cells became nonadherent and together with the added effector cells were removed by washing. The remaining viable adherent target cells were stained with acrydine orange contained in the quencher and optically counted by microcomputer. A notable dose-dependent killing on target tumor cells was reproducibly obtained by tested effector cells. Cytotoxic activities of most effector cells were significantly higher in the 24-h ATD assay than those of concordant 4-h target cell lysis test. Chloroquine (chq) inhibition test in the 24-h ATD assay showed negative or weak inhibition on cytotoxicity against adherent targets. Linear regression analysis also manifested the lack of a close correlation between 4- and 24-h target cell lysis tests, indicating these two assays measured different killing activities of the same effector cells. Because of its high sensitivity and reproducibility, this semiautomatic 24-h assay with computerized fluorescence measurement system could serve as a sensitive screening assay to select high cytotoxic tumor killer cells in adoptive immunotherapy.