Pineal cysts may promote pubertal development in girls with central precocious puberty: a single-center study from China.

Introduction: Pineal cysts have long been considered a benign intracranial variation. However, in our clinical practice, it has been observed that some children with central precocious puberty (CPP) who have pineal cysts experience rapid progression in adolescent development. In recent years, there has been a significant increase in the prevalence of CPP in girls, leading to more diagnoses of CPP among children with pineal cysts. Despite this, there is no consensus regarding whether pineal cysts contribute to CPP as one of its organic factors. This study aimed to analyze the clinical characteristics of pineal cysts in children with CPP and explore the potential effects of pineal cysts on puberty development. Methods: This single-center study retrospectively analyzed clinical data from girls aged 3 to 10 years who underwent head/pituitary magnetic resonance imaging at the Children's Hospital Affiliated to Zhengzhou University between 2019 and 2022. The study categorized the detection rates of pineal cysts based on systematic disease classification and compared the rates of cyst detection between girls diagnosed with CPP and those without CPP. Subsequently, CPP-diagnosed girls with pineal cysts were examined. Among CPP-diagnosed girls meeting the study's criteria, those with pineal cysts formed the 'cyst group,' while those without cysts were matched in a 1:1 ratio based on age and body mass index to form the 'non-cyst group.' Comparative analyses were conducted to assess the clinical characteristics between these two groups. CPP-diagnosed girls with cysts were further subdivided into three groups according to cyst size (≤5 mm, 5.1-9.9 mm, and ≥10 mm) to investigate potential differences in clinical characteristics among these subgroups. The study involved an analysis of clinical data from girls diagnosed with CPP and included imaging follow-ups to explore the progression of pineal cysts over time. Results: Among the 23,245 girls who underwent head/pituitary magnetic resonance imaging scans, the detection rate of pineal cysts was 3.6% (837/23,245), with most cases being associated with endocrine diseases. The detection rate of pineal cysts in CPP patients was 6.4% (262/4099), which was significantly higher than the 3.0% (575/19,146) in patients without CPP. In comparison to the non-cyst group, the cyst group exhibited statistically significant increases in estradiol levels, peak luteinizing hormone (LH) levels, peak LH/follicle-stimulating hormone (FSH) ratios, uterine body length, and cervix length (P < 0.001). As cyst size increased, there were significant rises in LH peak, peak LH/FSH ratio, uterine body length, and cervical length (P < 0.01). Estradiol levels and left ovarian volume also showed an increasing trend (P < 0.05). Among girls who underwent follow-up imaging, 26.3% (5/19) exhibited an increase in cyst size. Conclusion: Pineal cysts are relatively common in children with CPP. They may affect the pubertal development process, with larger cysts correlating to faster pubertal development. Therefore, the authors hypothesize that pineal cysts may trigger CPP in some cases, especially when the cysts are larger than 5 mm in size, as indicated by our data.

Serum vitamin B12 is a promising auxiliary index for the diagnosis of methylmalonic acidemia in children: A single center study in China.

BACKGROUND AND AIMS: Vitamin B12 (cobalamin, VitB12) is an essential coenzyme of methylmalonyl-CoA mutase and methionine synthase. Variations in VitB12 metabolism, absorption, transport, or intake may cause changes in methylmalonic acidemia (MMA) biomarkers. We aimed to investigate whether serum Vitamin B12 levels could be used in the early detection of MMA. MATERIALS AND METHODS: We included 241 children with MMA and 241 healthy matched controls. We measured serum VitB12 levels by an enzyme immunoassay and investigated the relationship between abnormal VitB12 levels and hematologic parameters as potential risk factors for MMA symptoms. RESULTS: Compared with controls, the serum levels of VitB12 were increased in the MMA group (p < 0.001). Serum VitB12 distinguished patients with MMA from healthy children (p < 0.001). Serum VitB12 combined with homocysteine and ammonia identified cblC and mut type MMA, respectively (p < 0.001). Homocysteine, folate, ammonia, NLR, and red blood cells contributed to serum VitB12 in cblC type MMA (p < 0.001); homocysteine, ammonia, and red blood cells, contributed in mut type MMA (p < 0.001); and elevated VitB12 was an independent predictor of MMA clinical onset (p < 0.001). CONCLUSION: Serum VitB12 can be used as an early diagnostic biomarker for MMA in children.

[Genetic analysis of a case with 11ß hydroxylase deficiency caused by CYP11B2/CYP11B1 chimeric gene].

OBJECTIVE: To analyze a child with 11ß hydroxylase deficiency (11ß-OHD) due to CYP11B2/CYP11B1 chimeric gene. METHODS: Clinical data of the child who was admitted to Henan Children's Hospital on August 24, 2020 were retrospectively analyzed. Peripheral blood samples of the child and his parents were collected and subjected to whole exome sequencing (WES). Candidate variant was verified by Sanger sequencing. RT-PCR and Long-PCR were carried out to verify the presence of chimeric gene. RESULTS: The patient, a 5-year-old male, had featured premature development of secondary sex characteristics and accelerated growth, and was diagnosed with 21 hydroxylase deficiency (21-OHD). WES revealed that he has harbored a heterozygous c.1385T>C (p.L462P) variant of the CYP11B1 gene, in addition to a 37.02 kb deletion on 8q24.3. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the c.1385T>C (p.L462P) was rated as a likely pathogenic variant (PM2_Supporting+PP3_Moderate+PM3+PP4). The results of RT-PCR and Long-PCR suggested that CYP11B1 and CYP11B2 genes have recombined to form a CYP11B2 exon 1~7/CYP11B1 exon 7~9 chimeric gene. The patient was diagnosed as 11ß-OHD and effectively treated with hydrocortisone and triptorelin. A healthy fetus was delivered following genetic counseling and prenatal diagnosis. CONCLUSION: 11ß-OHD may be misdiagnosed as 21-OHD due to the potential CYP11B2/CYP11B1 chimeric gene, which will require multiple methods for the detection.

Laser ultrasonic imaging for defect detection on metal additive manufacturing components with rough surfaces.

Defects or discontinuities are inevitable during the melting and consolidation process of metal additive manufacturing. Online inspection of microdefects during the processing of layer-by-layer fusion is urgently needed for quality control. In this study, the laser ultrasonic C-scan imaging system is established to detect the surface defects of selective laser melting (SLM) samples that have a different surface roughness. An autosizing method based on the maximum correlation coefficient and lag time is proposed to accurately measure the defect length. The influences of the surface roughness on the laser ultrasound signal-to-noise ratio distribution and defect sizing accuracy are also studied. The results indicate that the proposed system can detect notches with a depth of 50 µm and holes with a diameter of 50 µm, comparable in size to raw powder particles. The average error for the length measurement can reach 1.5% if the notch is larger than 2 mm. Meanwhile, the sizing error of a 1 mm length notch is about 9%. In addition, there is no need to remove the rough surface of the as-built SLM samples during the detection process. Hence, we propose that the laser ultrasonic imaging system is a potential method for online inspection of metal additive manufacturing.

The deubiquitinase USP34 stabilizes SOX2 and induces cell survival and drug resistance in laryngeal squamous cell carcinoma.

Recent studies showed that the deubiquitinase ubiquitin-specific protease 34 (USP34) was involved in the tumorigenesis of several tumors, but its function and mechanism are still unclear in laryngeal squamous cell carcinoma (LSCC). In this study, we found that USP34 and SOX2 were elevated in LSCC tumor tissues, and we also found that USP34 expression was positively correlated with SOX2 expression. Our further studies showed that USP34 regulated the protein level of SOX2 in LSCC cells, but not the mRNA level, which suggested that USP34 stabilized SOX2. Moreover, USP34, as a deubiquitinase, could interact with SOX2, and reduce the polyubiquitination of SOX2. In addition, knockdown of USP34 could significantly inhibit LSCC cell growth, but overexpression of SOX2 could reverse this effect. Finally, we also found that USP34 and SOX2 were upregulated in cisplatin-resistant LSCC cells, but knockdown of USP34 could enhance the drug sensitivity of cisplatin in the resistant cells. Collectively, targeting USP34/SOX2 axis may be a promising strategy for the treatment of LSCC.

A Novel Anti-Microbial Peptide from Pseudomonas, REDLK Induced Growth Inhibition of Leishmania tarentolae Promastigote In Vitro.

Leishmaniasis is a prevalent cause of death and animal morbidity in underdeveloped countries of endemic area. However, there is few vaccine and effective drugs. Antimicrobial peptides are involved in the innate immune response in many organisms and are being developed as novel drugs against parasitic infections. In the present study, we synthesized a 5-amino acid peptide REDLK, which mutated the C-terminus of Pseudomonas exotoxin, to identify its effect on the Leishmania tarentolae. Promastigotes were incubated with different concentration of REDLK peptide, and the viability of parasite was assessed using MTT and Trypan blue dye. Morphologic damage of Leishmania was analyzed by light and electron microscopy. Cellular apoptosis was observed using the annexin V-FITC/PI apoptosis detection kit, mitochondrial membrane potential assay kit and flow cytometry. Our results showed that Leishmania tarentolae was susceptible to REDLK in a dose-dependent manner, disrupt the surface membrane integrity and caused parasite apoptosis. In our study, we demonstrated the leishmanicidal activity of an antimicrobial peptide REDLK from Pseudomonas aeruginosa against Leishmania tarentolae in vitro and present a foundation for further research of anti-leishmanial drugs.

Highly active electrocatalysts of iron phthalocyanine by MOFs for oxygen reduction reaction under alkaline solution.

Metal-nitrogen-carbon materials (Fe-N/C) have been extensively studied as one of the most excellent electrocatalysts with good catalytic activities and cheap price towards the oxygen reduction reaction (ORR). The rational design of metal-organic framework (MOF) derived carbon materials with rapid mass transport ability and good stability is a great challenge to achieve. Herein, intensive research of Fe-N/C catalysts prepared from assembling MOFs with cheap iron phthalocyanine (FePc) for the ORR is innovatively carried out. A series of Fe-N/C nano-architectures are simply synthesized by a convenient assembling method under different temperatures (800 to 1000 °C). The assembly method at high temperatures tunes the number of FeN x active sites and intensifies the exposure of interior active sites. The highly dispersing Fe20-N/C electrocatalyst treated at 900 °C exhibits remarkable stability and excellent ORR activities with a half-wave potential of 0.866 V (vs. RHE) in alkaline solution, which is higher than that of commercial Pt/C (0.838 V vs. RHE) under the same test conditions. X-ray photoelectron spectroscopy results illustrate that incorporated MOFs interact with the active centre of FePc, tend to enhance the electron transition and to promote the kinetics of the ORR. Overall, highly dispersed Fe-N/C MOF-based materials are excellent non-precious metal electrocatalysts for energy and environmental applications.

Enhanced Enzymatic Hydrolysis and Lignin Extraction of Wheat Straw by Triethylbenzyl Ammonium Chloride/Lactic Acid-Based Deep Eutectic Solvent Pretreatment.

Efficient and feasible pretreatment of lignocellulosic biomass waste is an important prerequisite step to promote subsequent enzymatic hydrolysis and enhance the economics of biofuels production. This study focuses on the pretreatment of wheat straw (WS) with triethylbenzyl ammonium chloride/lactic acid (TEBAC/LA)-based deep eutectic solvents to enhance biomass fractionation and lignin extraction. Effects of pretreatment time, temperature, and TEBAC/LA molar ratio on pretreatment were evaluated systematically. Results suggested that 89.06 ± 1.05% of cellulose and 71.00 ± 1.03% of xylan were hydrolyzed with enzyme loadings of 35 FPU cellulase and 82 CBU ß-glucosidase (per gram of dry biomass) after pretreatment by TEBAC/LA (1:9) at 373 K for 10 h. A total monosaccharide yield of 0.550 g/g WS (91.27% of the theoretical yield) was achieved with 79.73 ± 0.93% of lignin removal. Furthermore, the 1H-13C two-dimensional heteronuclear single quantum correlation (2D-HSQC) NMR spectroscopy showed that the regenerated lignin (75.69 ± 1.32% purity) was mainly composed of the syringyl units and the guaiacyl units. Overall, the results in this study provide an effective and facile pretreatment method for lignocellulosic biomass waste to enhance enzymatic hydrolysis saccharification.

Seroprevalence of Dirofilaria immitis in Cats from Liaoning Province, Northeastern China.

The present study was performed to investigate the seroprevalence and risk factors for Dirofilaria immitis infection in cats from Liaoning province, northeastern China. From October 2014 to September 2016, sera of 651 cats, including 364 domestic cats and 287 feral cats (332 females and 319 males) were assessed. They were tested for the presence of D. immitis antigen using SNAP Heartworm RT test kit. In this population, the average prevalence was 4.5%. Age and rearing conditions (feral or domestic) were found to be associated with the prevalence of D. immitis. The prevalence was significantly higher in feral cats compared with domestic cats (8.4% vs 1.4%, P<0.01). There was no significant difference between males and females (4.7% vs 4.2%, P>0.05), but older cats (≥3 years old) showed a statistically higher prevalence compared with younger cats (<3 years old) in feral populations (16.8 vs 2.4%, P<0.01), while the difference between the age groups was not statistically significant in domestic cats (2.4% vs 0.51%, P>0.05), all these results suggest that outdoor exposure time may be one of the most important factors for D. immitis prevalence in cats. Results reveal that D. immitis are prevalence in domestic and feral cats in northeastern China, which indicates that appropriate preventive measures should be taken to decrease the incidence of feline heartworm disease in Liaoning province, northeastern China.

Toxoplasma gondii: Protective immunity induced by a DNA vaccine expressing GRA1 and MIC3 against toxoplasmosis in BALB/c mice.

The intracellular parasite Toxoplasma gondii is a major cause of abortion and neonatal loss in livestock, and can cause severe illness to human with weakened immune system. The heavy incidence and severe consequence indicate the development of vaccines against T. gondii is required. In this study, DNA vaccines encoding GRA1 and MIC3 antigens were developed. The parasite-specific immune responses and protection efficiency against toxoplasmosis by these DNA vaccines were evaluated in BALB/c mice. The results demonstrated that the IgG antibody production was significantly increased in multi-antigenic vaccine encoding GRA1 and MIC3 immunized group, as well as the IFN-γ level, when compared with single-gene vaccines and controls groups (p < 0.05). Two weeks after the final vaccination, the mice were challenged with either 1 × 10(4) or 1 × 10(2) RH strain tachyzoites, and the mortality and parasite reduction were observed. The multi-antigenic vaccine encoding GRA1 and MIC3 lead to the longest survival time as well as the less parasite-loads in brain and liver of immunized mice (p < 0.01). The present study indicates that the GRA1 and MIC3 showed the potential as target for vaccine investigation against toxoplasmosis. And the immune efficacy induced by multi-antigenic vaccine encoding GRA1 and MIC3 was better than that induced by single-antigenic vaccines alone.

Establishment and application of a loop-mediated isothermal amplification method for simple, specific, sensitive and rapid detection of Toxoplasma gondii.

The Loop-mediated isothermal amplification (LAMP) method amplifies DNA with high simply, specificity, sensitivity and rapidity. In this study, A LAMP assay with 6 primers targeting a highly conserved region of the GRA1 gene was developed to diagnose Toxoplasma gondii. The reaction time of the LAMP assay was shortened to 30 min after optimizing the reaction system. The LAMP assay was found to be highly specific and stable. The detection limit of the LAMP assay was 10 copies, the same as that of the conventional PCR. We used the LAMP assay to develop a real-time fluorogenic protocol to quantitate T. gondii DNA and generated a log-linear regression plot by plotting the time-to-threshold values against genomic equivalent copies. Furthermore, the LAMP assay was applied to detect T. gondii DNA in 423 blood samples and 380 lymph node samples from 10 pig farms, and positive results were obtained for 7.8% and 8.2% of samples, respectively. The results showed that the LAMP method is slightly more sensitive than conventional PCR (6.1% and 7.6%). Positive samples obtained from 6 pig farms. The LAMP assay established in this study resulted in simple, specific, sensitive and rapid detection of T. gondii DNA and is expected to play an important role in clinical detection of T. gondii.