[Effect of complete Y chromosome AZFc microdeletion on embryo euploidy].

OBJECTIVE: Preimplantation genetic testing (PGT) was performed to analyze the embryo euploidy in patients with complete Y chromosome AZFc microdeletion. METHODS: The clinical data of complete AZFc microdeletion underwent PGT from January 2013 to December 2021 in Reproductive Medicine Center of the First Affiliated Hospital of Nanjing Medical University were retrospectively analyzed. The patients with monogenic disease who underwent PGT during the same period were set as the control group. The basic characteristics, fertilization rate, Day 3 high quality embryo rate, blastocyst formation rate, embryo euploid rate, 45, X embryo ratio was compared between the two groups. RESULTS: A total of 220 patients were included, including 91 patients with complete AZFc microdeletion and 129 patients with monogenic disease. There was no significant difference in age between the two groups. In semen parameters, the sperm concentration, total sperm count and progressive motility in AZFc microdeletion group were lower than those in monogenic disease group, and the differences were statistically significant (P=0.001). The fertilization rate of AZFc microdeletion group was lower than that in monogenic disease group (P=0.012), and there was no significant difference in the number of MII oocytes, Day 3 high-quality embryo rate and blastocyst formation rate. A total of 933 blastocysts were successfully tested, including 496 blastocysts in AZFc deletion group and 437 blastocysts in monogenic disease group. The euploid, aneuploid and mosaic rates of the AZFc microdeletion group were 57.26%, 24.60% and 18.14%, respectively, while those of the monogenic disease group were 66.13%, 23.80% and 10.07%, with statistically significant differences between the two groups (P=0.001). Further analysis of the two groups of aneuploid embryos showed that aberrations occurred most commonly in chromosome16 (3.87%), X (3.46%), 13 (2.44%), 22 (2.24%) and 19 (2.03%) in AZFc microdeletion group, respectively, while the monogenic disease group was 22 (4.35%), 16 (2.97%), 7 (2.74%), 15(1.60%) and 2(1.60%), respectively. The proportion of sex chromosome abnormality in AZFc microdeletion group was higher than that in monogenic disease group (P=0.039), and there was no significant difference in the proportion of 45,X between the two groups. CONCLUSION: Compared with monogenic disease group, The embryo euploid rate in AZFc microdeletion patients decreased and the proportion of 45, X embryos did not increase significantly. It is recommended to select euploid female embryos by PGT, which not only avoids vertical transmission of AZFc microdeletion, but also reduces the risk of miscarriage due to aneuploid embryos.

[Characteristics and Reactivity of Ambient VOCs in Urban Hangzhou, China].

The ambient concentration of 122 volatile organic compound (VOC) species were continuously measured in urban Hangzhou, China from May 2018 to April 2019. The average mixing ratio of VOCs was (59.4±23.6)×10-9 and the oxygenated VOCs (OVOC) were the largest component. There was no clear "weekend effect" in urban Hangzhou, while the concentration of VOCs had a sharp decrease during long holidays. The concentration of VOCs had a positive correlation with air quality index (AQI) and reached the highest level when the primary pollutant was PM2.5. The assessment results of atmospheric chemical reactivity with·OH radical loss rate (L·OH) and ozone formation potential (OFP) showed the average value of L·OH was 7.5 s-1 and that of OFP was 152.1×10-9, among which carbonyl compounds, aromatics, and alkenes were the most abundant components. The overall chemical reactivity level of VOCs in Hangzhou was equivalent to 2-methylpentane. The average value of toluene/benzene (T/B) was 1.95, which implied the ambient VOCs in Hangzhou were influenced by vehicle exhaust. Secondary formation (17.6%), combustion (11.8%), industrial processing (12.3%), solvent use (18.1%), biogenic source (4.5%), and vehicle exhaust (35.7%) were identified as six major sources of VOCs in Hangzhou through the positive matrix factorization (PMF) model.

[Characteristics and Reactivity of VOCs in Hangzhou During a Typical Photochemical Pollution Episode].

An intensive observation of ambient volatile organic compounds (VOCs) was carried out in Hangzhou, a key city in the Yangtze River Delta, during a typical photochemical pollution episode from September 14-23, 2018. The analysis results of 80 effective samples showed that the average concentration of 122 compounds of VOCs was (59.5±19.8)×10-9 during the observation period, and oxygenated VOCs (OVOCs) were the most abundant component. The assessment results of atmospheric reaction activity with ozone formation potential (OFP) showed that the average value of OFP was 145×10-9 during the observation period, of which alkenes and carbonyl compounds were the most abundant components. The chemical reactivity of VOCs in Hangzhou was equivalent to acrylonitrile. Based on the positive matrix factorization (PMF) model, five major sources of VOCs in Hangzhou were identified, including secondary formation (25.2%), combustion and industrial processing (27.2%), solvent use (17.3%), biogenic sources (9.2%), and vehicular exhaust (21.2%). The results can provide guidance for further understanding of VOC characteristics and the basis for scientific prevention and control measures in Hangzhou.

[Preimplantation genetic diagnosis for infertile males with autosomal dominant polycystic kidney disease].

OBJECTIVE: Autosomal dominant polycystic kidney disease (ADPKD) is one of the most common genetic renal diseases, which may cause oligoasthenospermia and azoospermia and result in male infertility. This study aimed to analyze the outcomes of preimplantation genetic diagnosis (PGD) in male patients with ADPKD-induced infertility. METHODS: We retrospectively analyzed the clinical data on 7 male patients with ADPKD-induced infertility undergoing PGD from April 2015 to February 2017, including 6 cases of oligoasthenospermia and 1 case of obstructive azoospermia, all with the PKD1 gene heterozygous mutations. Following intracytoplasmic sperm injection (ICSI), we performed blastomere biopsy after 5 or 6 days of embryo culture and subjected the blastomeres to Sureplex whole-genome amplification, followed by haplotype linkage analysis, Sanger sequencing, array-based comparative genomic hybridization to assess the chromosomal ploidy of the unaffected embryos, and identification of the unaffected euploid embryos for transfer. RESULTS: One PGD cycle was completed for each of the 7 patients. Totally, 26 blastocysts were developed, of which 12 were unaffected and diploid. Clinical pregnancies were achieved in 6 cases following 7 cycles of frozen embryo transplantation, which included 5 live births and 1 spontaneous abortion. CONCLUSIONS: For males with ADPKD-induced infertility, PGD may contribute to high rates of clinical pregnancy and live birth and prevent ADPKD in the offspring as well. This finding is also meaningful for the ADPKD patients with normal fertility.

High expression of ubiquitin-conjugating enzyme E2A predicts poor prognosis in hepatocellular carcinoma.

The present study aimed to illustrate the association of the expression of ubiquitin-conjugating enzyme E2A (UBE2A) with the clinicopathological parameters and prognosis in hepatocellular carcinoma (HCC). The expression levels of UBE2A mRNA and protein in a total of 276 HCC tissues and six liver cell lines was detected by fluorescent quantitative polymerase chain reaction, western blotting and immunohistochemistry. Statistical analysis was also performed to assess the association of the expression of UBE2A with the clinicopathological parameters and prognosis by the GraphPad Prism and SPSS version 21.0 software. UBE2A mRNA and protein were highly expressed in HCC tissues compared with those in the adjacent normal tissue. Immunohistochemical analysis revealed that UBE2A protein was more strongly stained in the 276 paraffin-embedded HCC tissues as compared with the 63 adjacent normal tissue. Statistical analysis also demonstrated that UBE2A expression was significantly associated with histological differentiation, TNM stage and vascular invasion of HCC (P<0.05). Notably, HCC patients with a high expression of UBE2A had a shorter survival time as compared with those with a low expression of UBE2A. There results suggested that UBE2A may be involved in the pathogenesis of HCC and may serve as an important prognostic marker. Further exploration of the involvement of UBE2A in HCC development may provide novel therapeutic targets.

Low expression of trafficking protein particle complex 4 predicts poor prognosis in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide. Tumor recurrence and metastasis are major factors that contribute to the poor outcome of patients with HCC. However, it is difficult to predict the prognosis of hepatocellular carcinoma. Trafficking Protein Particle Complex 4 (Trappc4), is associated with tumorigenesis. The present study aimed to detect Trappc4 expression in HCC and its association with clinicopathological patient data. More importantly, this study reveals the relationship between Trappc4 and the prognosis of hepatocellular carcinoma. A total of 148 HCC tissues were assessed for expression of Trappc4 mRNA and protein with (reverse transcription polymerase chain reaction) RT-PCR (n=36), Western blotting (n=4) and immunohistochemistry (n=148), respectively. The data show that Trappc4 mRNA and protein are expressed at low levels in HCC tissues compared to adjacent tissues. Immunohistochemical analysis revealed that 148 cases of HCC showed different degrees of positive expression. Statistical analysis showed that expression of Trappc4 was associated with histological differentiation, TNM stage, and vascular invasion (P < 0.05), but did not correlate with the patient's age, gender, tumor size (P > 0.05). Most importantly, HCC patients with low expression of Trappc4 had shorter survival time compared to patients with high expression. Trappc4 might be involved in the pathogenesis of HCC and could be an important prognostic marker in HCC patients.

Cathepsin L is an immune-related protein in Pacific abalone (Haliotis discus hannai)--Purification and characterization.

Cathepsin L, an immune-related protein, was purified from the hepatopancreas of Pacific abalone (Haliotis discus hannai) by ammonium sulfate precipitation and column chromatographies of SP-Sepharose and Sephacryl S-200 HR. Purified cathepsin L appeared as two bands with molecular masses of 28.0 and 28.5 kDa (namely cathepsin La and Lb) on SDS-PAGE under reducing conditions, suggesting that it is a glycoprotein. Peptide mass fingerprinting (PMF) analysis revealed that peptide fragments of 95 amino acid residues was high similarity to cathepsin L of pearl oyster (Pinctada fucata). The optimal temperature and pH of cathepsin L were 35 °C and pH 5.5. Cathepsin L was particularly inhibited by cysteine proteinase inhibitors of E-64 and leupeptin, while it was activated by metalloproteinase inhibitors EDTA and EGTA. The full-length cathepsin L cDNA was further cloned from the hepatopancreas by rapid PCR amplification of cDNA ends (RACE). The open reading frame of the enzyme was 981 bp, encoding 327 amino acid residues, with a conserved catalytic triad (Cys134, His273 and Asn293), a potential N-glycosylation site and conserved ERFNIN, GNYD, and GCGG motifs, which are characteristics of cathepsin L. Western blot and proteinase activity analysis revealed that the expression and enzyme activity of cathepsin L were significantly up-regulated in hepatopancreas at 8 h following Vibrio parahaemolyticus infection, demonstrating that cathepsin L is involved in the innate immune system of abalone. Our present study for the first time reported the purification, characterization, molecular cloning, and tissue expression of cathepsin L in abalone.

[Light scattering extinction properties of atmospheric particle and pollution characteristics in hazy weather in Hangzhou].

In order to evaluate the influence of particle scattering on visibility, light scattering coefficient, particle concentrations and meteorological factor were simultaneously monitored from July 2011 to June 2012 in Hangzhou. Daily scattering coefficients ranged from 108.4 to 1 098.1 Mm(-1), with an annual average concentration of 428.6 Mm(-1) ± 200.2 Mm(-1). Seasonal variation of scattering coefficients was significant, with the highest concentrations observed in autumn and winter and the lowest in summer. It was found there were two peaks for the average diurnal variations of the scattering coefficient, which could be observed at 08:00 and 21:00. The scattering efficiencies of PM2.5 and PM10 were 7.6 m2 x g(-1) and 4.4 m2 x g(-1), respectively. The particle scattering was about 90.2 percent of the total light extinction. The scattering coefficients were 684.4 Mm(-1) ± 218.1 Mm(-1) and 1 095.4 Mm(-1) ± 397.7 Mm(-1) in hazy and heavy hazy days, respectively, which were 2.6 and 4.2 times as high as in non-hazy weather, indicating that particle scattering is the main factor for visibility degradation and the occurrence of hazy weather in Hangzhou.