Exosomal Vaccine Loading T Cell Epitope Peptides of SARS-CoV-2 Induces Robust CD8+ T Cell Response in HLA-A Transgenic Mice.

Purpose: Current vaccines for the SARS-CoV-2 virus mainly induce neutralizing antibodies but overlook the T cell responses. This study aims to generate an exosomal vaccine carrying T cell epitope peptides of SARS-CoV-2 for the induction of CD8+ T cell response. Methods: Thirty-one peptides presented by HLA-A0201 molecule were conjugated to the DMPE-PEG-NHS molecules, and mixed with DSPE-PEG to form the peptide-PEG-lipid micelles, then fused with exosomes to generate the exosomal vaccine, followed by purification using size-exclusion chromatography and validation by Western blotting, liquid nuclear magnetic resonance (NMR) test and transmission electron microscopy. Furthermore, the exosomal vaccine was mixed with Poly (I:C) adjuvant and subcutaneously administered for three times into the hybrid mice of HLA-A0201/DR1 transgenic mice with wild-type mice. Then, the epitope-specific T cell responses were detected by ex vivo ELISPOT assay and intracellular cytokine staining. Results: The exosomal vaccine was purified from the Peak 2 fraction of FPLC and injected into the hybrid mice for three times. The IFN-γ spot forming units and the frequencies of IFN-γ+/CD8+ T cells were 10-82-fold and 13-65-fold, respectively, higher in the exosomal vaccine group compared to the Poly (I:C) control group, without visible organ toxicity. In comparison with the peptides cocktail vaccine generated in our recent work, the exosomal vaccine induced significantly stronger T cell response. Conclusion: Exosomal vaccine loading T cell epitope peptides of SARS-CoV-2 virus was initially generated without pre-modification for both peptides and exosomes, and elicited robust CD8+ T cell response in HLA-A transgenic mice.

Effect of Mirena placement on reproductive hormone levels at different time intervals after artificial abortion.

BACKGROUND: Improper methods of contraception greatly increase the risk of abortion, cervical or endometrial lesions, and the number of recurrent artificial abortions. These complications result in the deterioration of a patient's outcome. Further, the proportion of artificial abortions is highest among unmarried females. Placement of an intrauterine device, such as the Mirena, after an artificial abortion may decrease the likelihood of an endometrial injury caused by recurrent abortions while significantly improving its contraceptive effects. AIM: To discuss the effect of Mirena placement on reproductive hormone levels at different time points after an artificial abortion. METHODS: Women (n = 119) undergoing an artificial abortion operation were divided into the study (n = 56) and control (n = 63) groups. In the study group, the Mirena was inserted immediately after the artificial abortion, whereas in the control group, it was inserted 4-7 d after the onset of the first menstrual cycle after abortion. All participants were followed-up for 6 mo to observe the continuation and expulsion rates and adverse reactions and to measure the levels of serum estradiol (E2), follicle stimulating hormone (FSH), and luteinizing hormone (LH). RESULTS: The continuation rates were 94.64% and 93.65% in the study group and the control group, respectively. The expulsion rates were 1.79% and 3.17% in the study group and the control group, respectively. There was no statistically significant difference between the two groups (P > 0.05). There were also no statistically significant differences in the proportion of patients with bacterial vaginitis, trichomonas vaginitis, or cervicitis between the groups (P > 0.05). Six months after Mirena placement, E2 Levels were 45.50 ± 7.13 pg/mL and 42.91 ± 8.10 pg/mL, FSH 13.60 ± 3.24 mIU/mL and 14.54 ± 3.11 mIU/mL, and LH 15.11 ± 2.08 mIU/mL and 14.60 ± 3.55 mIU/mL in the study and control groups, respectively. There were no significant differences in hormone levels between the two groups (P > 0.05). There were also no statistically significant differences in the proportions of abnormal menstruation, prolonged menstruation, or pain during intercourse between the study and control groups after Mirena placement (P > 0.05). There were no statistically significant differences in uterine volume, sexual desire, sexual activity, or the sexual satisfaction score between the study and control groups before and after Mirena placement (P > 0.05). CONCLUSION: Placement of a Mirena intrauterine device immediately after an artificial abortion does not increase the risk of adverse reactions and can help prevent endometrial injury caused by recurrent abortions.

Enhancing the differentiation of walking and standing via the ratio of plantar pressures.

Differentiation of standing and walking based on plantar pressures is helpful in developing strategies to reduce health risks in the workplace. In order to improve the differentiation ability, the paper proposes a new metric for posture differentiation, that is, the pressure ratio on the two anatomical plantar regions. The plantar pressures were collected from 30 persons during walking and standing. After verifying the normal distribution of the pressure ratio by the Monte Carlo method, two-way repeated-measures ANOVA was conducted for the pressure ratios. The advantage of the pressure ratio over two conventional pressure metrics (the average pressure and the peak pressure) is demonstrated by its much larger size effect. Furthermore, the pressure ratio permits to establish value ranges corresponding to walking and standing, which are less influenced by specific person factors, thus facilitating the design of a standardized posture recognition system. The underlying mechanism underlying the pressure ratio is discussed from the aspect of biomechanics of movement.

Clinical Application of Noninvasive Prenatal Testing for Pregnant Women with Assisted Reproductive Pregnancy.

OBJECTIVE: This paper analyzes the clinical significance of noninvasive prenatal testing (NIPT) for fetal chromosome aneuploidy in the screening of in vitro fertilization-embryo transfer (IVF) pregnancies. METHODS: The study subjects consisted of 3163 IVF-pregnant women who underwent NIPT at the Women's Hospital, School of Medicine, Zhejiang University and Taizhou Hospital, Zhejiang Province from February 2015 to June 2019. Fetal or neonatal karyotype analysis was carried out in high-risk patients, with subsequent follow-up on pregnancy outcomes. RESULTS: NIPT results of 3163 pregnant women suggested 20 cases of high-risk fetal chromosome aneuploidy, of which 2185 cases were a single pregnancy. Of the 13 cases of high-risk chromosome aneuploidy in single pregnancies, seven were true positive, and six were false positive according to fetal or newborn chromosomal karyotype diagnosis. Twin pregnancies accounted for 978 cases in which NIPT indicated seven cases of high-risk chromosome aneuploidy; six of these cases were true positive, and one case was false positive according to fetal or newborn chromosomal karyotype diagnosis. The specificity, positive predictive value, and false-positive rate of trisomy 21 syndrome in IVF single embryo NIPT were 99.86%, 62.5%, and 0.14%, respectively. The specificity, positive predictive value, and false-positive rate of trisomy 18 syndrome were 99.95%, 66.67%, and 0.05%, respectively. The specificity of trisomy 13 syndrome was 99.91%, and the false-positive rate was 0.09%. The specificity of trisomy 21 syndrome in IVF twin NIPT was 99.89%, the positive predictive value was 83.33%, and the false-positive rate was 0.11%. The specificity and positive predictive value of fetal trisomy 18 syndrome were 100.00%, and the false-positive rate of it were 0.00%. Sensitivity and false-negative rates were 100% in all cases. CONCLUSION: NIPT is an ideal prenatal test for IVF-pregnant women due to its high sensitivity and specificity in screening for fetal aneuploidy.

Galectin-1 expression in the serum and placenta of pregnant women with fetal growth restriction and its significance.

BACKGROUND: This study aims to investigate galectin-1 (Gal-1) expression in the serum and placenta of pregnant women with fetal growth restriction (FGR) and its significance. METHODS: Thirty-one pregnant women with single-birth FGR but without comorbidities, eight pregnant women with FGR and preeclampsia (PE), and eight pregnant women with FGR and gestational diabetes mellitus (GDM) were enrolled as the study group, while 20 pregnant women with normal singleton pregnancy in the same period were enrolled as the control group. The serum Gal-1 level was detected using an enzyme-linked immunosorbent assay (ELISA), and Gal-1 expression in the placenta was detected by western blot. RESULTS: The results revealed that, compared with the control group, the serum Gal-1 level decreased in the women with FGR without comorbidities, and the difference was statistically significant (P < 0.001). Compared with the control group, the difference in serum Gal-1 expression in the FGR-PE group was not statistically significant (P = 0.29). The peripheral serum Gal-1 level decreased in the FGR-GDM group compared with the control group, and the difference was statistically significant (P < 0.001). The serum Gal-1 level was positively correlated with birth weight (r2 = 0.172, P < 0.01). Compared with the control group, the Gal-1 expression level decreased in the placenta of the pregnant women with FGR without comorbidities (P < 0.05). CONCLUSIONS: Gal-1 exhibits low expression in the serum and placenta of pregnant women with FGR. In addition, Gal-1 may be involved in the pathogenesis of FGR and could represent a new diagnostic marker of the disease.

The first complete mitochondrial genome from Bostrychus genus (Bostrychus sinensis) and partitioned Bayesian analysis of Eleotridae fish phylogeny.

To understand the phylogenetic position of Bostrychus sinensis in Eleotridae and the phylogenetic relationships of the family, we determined the nucleotide sequence of the mitochondrial (mt) genome of Bostrychus sinensis. It is the first complete mitochondrial genome sequence of Bostrychus genus. The entire mtDNA sequence was 16508 bp in length with a standard set of 13 protein-coding genes, 22 transfer RNA genes (tRNAs), two ribosomal RNA genes (rRNAs) and a noncoding control region. The mitochondrial genome of B. sinensis had common features with those of other bony fishes with respect to gene arrangement, base composition, and tRNA structures. Phylogenetic hypotheses within Eleotridae fish have been controversial at the genus level. We used the mitochondrial cytochrome b (cytb) gene sequence to examine phylogenetic relationships of Eleotridae by using partitioned Bayesian method. When the specific models and parameter estimates were presumed for partitioning the total data, the harmonic mean -lnL was improved. The phylogenetic analysis supported the monophyly of Hypseleotris and Gobiomorphs. In addition, the Bostrychus were most closely related to Ophiocara, and the Philypnodon is also the sister to Microphlypnus, based on the current datasets. Further, extensive taxonomic sampling and more molecular information are needed to confirm the phylogenetic relationships in Eleotridae.

[Characteristics and phylogenetic analysis of mitochondrial genome in the gobies].

The vast number of species, small size and high variation of morphology make the morphological identification and classification of gobies very difficult. In this study, the complete mitochondrial genome (mitogenome) of 26 species of gobies was analyzed, aiming at accumulating the molecular information on the identification, classification and molecular evolution of gobies. The results showed that the gene composition and arrangement of mitogenome of gobies are similar to most vertebrates. Due to various degrees of repetitive sequences in the control region, the mitogenome of 26 gobies exhibits a great variation in length. The A+T content of the mitogenome is greater than 50% and the lowest frequency is for G among the four bases. Thirty-seven coding gene sequences were used to calculate the average Kimura 2-parameter genetic distance of 26 species of gobies. Acanthogobius hasta and A. ommaturus, Glossogobius olivaceus and G circumspectus were synonyms, respectively. By comparing the control region sequences of 26 gobies, the terminal associated sequences, central conserved sequence block and conserved sequence block were identified, respectively. Thirty-six coding gene sequences of 26 gobies were used to construct the phylogenetic tree and the results were different from the traditional morphological classification. The five subfamilies of Gobiidae were obviously evolved: Amblyopinae, Oxudercinae and Sicydiinae were clustered into a group and then formed a sister group with Gobionellinae; the fishes of Gobiinae had distant relationship with the four subfamilies and formed a group alone. Molecular clock analysis estimated that gobies probably originated in the late Eocene to Oligocene time and further evolved into modern characteristic gobies in the Miocene.