Sex-Inclined Piwi-Interacting RNAs in Serum Exosomes for Sex Determination in the Greater Amberjack (Seriola dumerili).

The greater amberjack (Seriola dumerili) is a gonochoristic fish with no sexual dimorphism in appearance, making sex identification difficult. Piwi-interacting RNAs (piRNAs) function in transposon silencing and gametogenesis and are involved in various physiological processes, including sex development and differentiation. Exosomal piRNAs can be indicators for the determination of sex and physiological status. In this study, four piRNAs were differentially expressed in both serum exosomes and gonads between male and female greater amberjack. Three piRNAs (piR-dre-32793, piR-dre-5797, and piR-dre-73318) were significantly up-regulated and piR-dre-332 was significantly down-regulated in serum exosomes and gonads of male fish, compared to female fish, consistent with the serum exosomal results. According to the relative expression of four marker piRNAs derived from the serum exosomes of greater amberjack, the highest relative expression of piR-dre-32793, piR-dre-5797, and piR-dre-73318 in seven female fish and that of piR-dre-332 in seven male fish can be used as the standard for sex determination. The method of sex identification can ascertain the sex of greater amberjack by blood collection from the living body, without sacrificing fish. The four piRNAs did not show sex-inclined expression in the hypothalamus, pituitary, heart, liver, intestine, and muscle tissue. A piRNA-target interaction network involving 32 piRNA-mRNA pairs was generated. Sex-related target genes were enriched in sex-related pathways, including oocyte meiosis, transforming growth factor-beta signaling pathway, progesterone-mediated oocyte maturation, and gonadotropin releasing hormone signaling pathway. These results provide a basis for sex determination in greater amberjack and improve our understanding of the mechanisms underlying sex development and differentiation in the species.

The feasibility of skin mucus replacing exosome as a pool for bacteria-infected markers development via comparative proteomic screening in teleost.

In fish, skin mucus forms a protective barrier between the body surface and the external water environment, thus providing the most direct and intuitive clues to monitor the subject's health condition. To explore the impact of the Vibrio harveyi pathogen on teleost, the proteome of epidermal mucus from control and sick Cynoglossus semilaevis were screened through iTRAQ followed with LC-MS/MS. 1531 credible proteins were obtained relating to structural, metabolic and immunological functions. 335 different expressed proteins (DEPs) were identified, with 166 up-regulated and 169 down-regulated in MS. 62 proteins were characterized, including 22 up-regulated proteins and 40 down-regulated proteins. Integrated analysis of DE-miRNAs and DEPs from miRomics and proteomics were conducted to show the indirect regulatory relationship. Comparative analysis of DEPs between mucus and exosomes demonstrated that exosomes contributed the most DEPs of all mucus DEPs. 125 proteins are DEPs only in exosomes, which presented minor difference in total mucus. Expression of Aminopeptidase (anpep), Calcium-transporting ATPase, Histone H2B and H2A confirmed implied fine discriminative power with infected C. semilaevis, among which Calcium-transporting ATPase and H2B also appeared in list of exosomal markers. This study might shed the light on effective biomarker digging at other extended screening scenarios.

Comparative piRNAs Profiles Give a Clue to Transgenerational Inheritance of Sex-Biased piRNAs in Cynoglossus semilaevis.

Piwi interacting RNAs (piRNAs) are involved in the epigenetic and post-transcriptional gene silencing of retrotransposons in germ line cells, especially in spermatogenesis. There are many related reports on model organisms, such as flies and mice. In fish, however, there are few studies on piRNAs. Cynoglossus semilaevis, a benthic warm water flatfish, with remarkable sexual dimorphism, especially the "pseudo males" with sex reversal, mating with normal females to produce viable offspring, is an ideal material for the study of sex development. Here, sperm piwi-interacting RNAs profiles of Cynoglossus semilaevis were characterized, comparing between male and pseudomale groups. Differential piRNAs were identified with their predicted and annotated targets. Attention was then focused on candidate piRNAs associated with sex development and methylation. We continued to compare the expression levels of 10 candidates differentially expressed piRNAs in F1 spermatozoa. Quantitative RT-PCR demonstrated that five of the ten piRNAs showed sex bias consistent with parental sequencing results, with four significantly higher expression level in sperm of five males offspring than that of pseudomales, while one piRNAs showed the opposite expression profile. The five signature piRNAs (piR-mmu-49600337, piR-mmu-95849, piR-xtr-7474223, piR-xtr-1790334, and piR-mmu-4491546) could be employed as male-specific molecular biomarkers for C. semilaevis. Besides, this study also implied the possibility of transgenerational inheritance of sex-biased piRNAs exiting in sperm of Cynoglossus semilaevis.

Mucus piRNAs profiles of Vibrio harveyi-infected Cynoglossus semilaevis: A hint for fish disease monitoring.

The half-smooth tongue sole, Cynoglossus semilaevis, is an important cultured flatfish species. Vibrio harveyi is a common pathogen to this fish, which may result in great economic loss to C. semilaevis culture industry. piRNAs, a non-coding RNAs with 26-32 nt, have been regarded as promising biomarkers for cancer diagnosis and fish diseases. Here, we extracted the RNA from mucus of C. semilaevis and constructed the differential expression profiles of piRNAs between the sick fish (MS) and healthy fish (MC). We identified 45,696 differentially expressed piRNAs including 22,735 up-regulated piRNAs and 22,961 down-regulated piRNAs in MS group compared with MC group. The GO enrichment and KEGG pathway enrichment analyses of the differential piRNAs were carried out. The result showed immunity-related target genes mainly involved in immune system process, response to stimulus, cell killing, immune system, infectious diseases and cell growth and death. The 10 most differentially expressed piRNAs were chosen to perform the qRT-PCR, while only seven piRNAs were consistent with the sequence result. Compared with MC group, the expression levels of piR-mmu-72173>piR-rno-62831>piR-xtr-704880, piR-dme-15546979, piR-mmu-49941660, piR-mmu-29283297 and piR-mmu-1758399 were significantly lower, and piR-gga-10574 and piR-gga-134812 were significantly higher in MS group. These piRNAs may be potential biomarkers during the V. harveyi infection of C. semilaevis. This study could provide a new method to identify the infection status of C. semilaevis and understand better about the innate and adaptive immune system in C. semilaevis during bacterial infection.

Nutritional risk and nutritional status in hospitalized older adults living with HIV in Shenzhen, China: a cross-sectional study.

BACKGROUND: Nutrition is a crucial factor that can impact morbidity and mortality in older people living with HIV (PLWH). Studies on nutritional risk and nutritional status in all age groups in PLWH have been conducted. However, few studies have focused on nutritional risk in older PLWH. This study aimed to describe the nutritional risk and nutritional status in older PLWH, and explore factors associated with nutritional risk and undernutrition status. METHODS: We conducted a cross-sectional study. We recruited participants aged 50 years or older from the Third People's Hospital of Shenzhen from January 2016 to May 2019. Nutritional risk and nutritional status were evaluated by the Nutritional Risk Screening 2002 (NRS 2002) tool, body mass index (BMI), albumin level, and prealbumin level on the first day of admission. Logistic regression models were used to identify the factors associated with undernutrition based on the BMI, albumin, and prealbumin criteria. RESULTS: A total of 196 older PLWH were included in the analysis. We found that 36% of hospitalized older PLWH had nutritional risk, and 12-56% of them had undernutrition based on the BMI, albumin, and prealbumin criteria. An increased nutritional risk score was associated with older age (ß = 0.265 CI [0.021, 0.096], P = 0.002), a higher viral load (ß = - 0.186 CI [- 0.620, - 0.037], P = 0.028), a lower BMI (ß = - 0.287 CI [- 0.217, - 0.058], P = 0.001), and a lower albumin level (ß = - 0.324 CI [- 8.896, - 1.230], P = 0.010). The CD4 count was associated with the prevalence of undernutrition based on the albumin criterion (OR = 15.637 CI [2.742, 89.178], P = 0.002). CONCLUSION: Our study indicated that nutritional screening, assessment, and management should be routinely performed in hospitalized older PLWH. HIV-specific measures should be used to assess nutritional risk, and albumin, BMI, and other assessments should be used in combination to identify undernutrition in older PLWH.

[Effect and safety of electroacupuncture analgesia at the latent stage of labor on the epidural analgesia].

OBJECTIVE: To investigate the effect of electroacupuncture (EA) analgesia at the latent stage of labor on epidural analgesia, labor outcome and neonatal score so as to provide the reference to the clinical labor analgesia. METHODS: According to the random number table, 104 primipara of vaginal delivery were divided into three groups, named an EA group (37 cases), a sham-EA group (36 cases) and an epidural block group (31 cases). In the EA group, when the cervix opened up by 1 cm, Han's acupoint nerve stimulation apparatus was attached to bilateral Hegu (LI4) and bilateral Sanyinjiao (SP6) to achieve EA analgesia till the active stage (the cervix up 3 cm). In the sham-EA group, the procedure was same as the EA group, but with the sham-stimulation. In the epidural block, EA was not used in intervention. When the cervix opened up 3 cm, the epidural block and the patient-controlled epidural analgesia were all adopted in each group. The score of visual analogue scale (VAS) and the dosage of Sufentanil and Ropivacaine in patient-controlled analgesia pump at the different time points were compared among the groups. Respectively, in 1 h of EA stimulation (T1), 2 h of EA stimulation (T2), at the moment of epidural block (T3) and the complete open of the cervix (T6), the venous blood sample was collected to determine the concentration of beta-endorphin (beta-EP), and the use time of oxytocin, the bleeding amount and the state of newborn were recorded. RESULTS: There was no statistical difference in immediate of EA stimulation (T0) among the groups in VAS score (P>0.05). But, in T1 to T6, VAS score in the EA group was reduced obviously as compared with the sham-EA group and the epidural block group separately (P<0.05). There was no statistical difference in the use time of oxytocin among the three groups (P>0.05). In the sham-EA group and the epidural block group, bleeding amount during labor and in 24 h after labor was all higher than that of the EA group (P<0.05). In each group, the concentration of beta-EP in T3 and T6 was increased obviously as compared with that in T1 and T2 respectively (P<0.05). In the EA group, the concentration of beta-EP in T1 to T3 and in T6 was higher than that in sham-EA group and the epidural block group respectively (P<0.05). The dosage of Ropivacaine and Sufentanil in the EA group was less than that in either the sham-EA group or the epidural block group (P<0.05). There was no significant difference in amniotic fluid turbidity rate and Apgar score among the three groups (P>0.05). CONCLUSION: Electroacupuncture analgesia at the latent stage of labor effectively relieves labor pain of primipara, increases the concentration of beta-EP, enhances the effect of epidural block analgesia, reduces labor bleeding and has no side effect on newborn. The combination of electroacupuncture analgesia and the epidural anesthesia in the latent stage is applicable in the whole process of labor.

[Effect of preemptive electroacupuncture on pain reactions in primiparous parturients during vaginal delivery].

OBJECTIVE: To investigate the effect of preemptive electroacupuncture (EA) plus epidural block (EB) on pain reactions and stress hormone levels in primiparous parturients during labor. METHODS: A total of 104 primiparous parturients undergoing vaginal delivery were randomly divided into PA group (n=31), sham EA group (n=36) and EA group (n=37). EA (2 Hz/100 Hz, a tolerable strength) was applied to bilateral Hegu (LI4) and Sanyinjiao (SP6) beginning from the latency of the 1st birth process when the orifice of uterus was opened to about 1 cm wider till the active stage when the orifice of the uterus opened about 3 cm wider. Patients in the sham EA group received shallow acupuncture needle stimulation (without cutaneous penetration of the needle tip, and without stimulation electric current output). All the patients of the 3 groups were given EB (1% Lidocaine, 0.05% Sufentanil and 0.1% Ropivacaine) and patient-controlled epidural analgesia (PCEA) when the orifice of the uterus opened about 3 cm wider. The pain severity was assessed by using Visual Analogue Scale (VAS) at time-points of immediately after EA, 1 h after EA, 2 h after EA, immediately after EB, 1 h after EB, 2 h after EB and full open of the uterus orifice. The contents of serum adrenocorticotrophic hormone (ACTH) and cortisol (COR) were assayed by using ELISA. RESULTS: There was no significant difference in the VAS score at the immediate time of post-EA among the three groups (P>0.05). Compared with the EB group, the VAS scores at all time-points except the immediate time of post-EA were significantly decreased in the EA group (P<0.05). One and 2 h after EB, and after full open of the uterus orifice, the VAS scores in the 3 groups were all evidently decreased (P<0.05), and those of the EA group were significantly lower than those of the sham EA and EB groups (P<0.05). The three groups had a significant increase in the levels of serum ACTH and COR at time-points of 1 and 2 h after EA, immediately after EB, and when the orifice of the uterus was fully opened (P<0.05), presenting a gradual increase of their level during the course of labor. Compared with the EB group, the levels of serum ACTH and COR contents were considerably decreased in the EA group (rather than in the sham EA group) (P<0.05). The dosages of Ropivacaine and Sufentanil, the total pressing times and the effective pressing times of PCEA pump were significantly lower in the EA group than those in the EB and sham EA groups (P<0.05). CONCLUSION: EA plus epidural analgesia, given during the latency period of labor, can effectively alleviate pain, inhibit stress response and reduce the dosage of anesthetics in primiparous parturients throughout the labor process.