Concentrations and influencing factors of 17 elements in placenta, cord blood, and maternal blood of women from an e-waste recycling area.

BACKGROUND: The effects of prenatal element exposure on mothers and fetuses have generated concern. Profiles of trace and toxic elements in biological material are urgently desired, especially for women who reside near e-waste recycling facilities. The aim of this study was to investigate elements concentrations in placenta, cord blood, and maternal blood of women and to evaluate the influencing factors. METHODS: A group of 48 women from an e-waste recycling site and a group of 31 women from a non-e-waste recycling site were recruited. Basic characteristics were collected by questionnaire and the concentrations of 17 elements in placenta, cord blood, and maternal blood samples were analyzed by inductively coupled plasma mass spectrometry (ICP-MS). Finally, the generalized linear model regression analysis (GLM) was used to test the association between element concentrations and possible factors. RESULTS: Compared to the control group, the exposed group had significantly elevated cadmium (Cd), zinc (Zn), nickel (Ni), and antimony (Sb) in placenta, and higher lead (Pb) in maternal blood and cord blood (P<0.05). Sb concentration in maternal blood was significantly lower than in the control group (P<0.05). GLM analysis showed that element concentrations were mainly associated with maternal age [chromium (Cr), iron (Fe), selenium (Se), cobalt (Co), mercury (Hg) in placenta, copper (Cu) in maternal blood], education (Se, Sb in placenta), family income (Cu in maternal blood and Ni in placenta), passive smoking [Cu and Zn in placenta, Pb in maternal blood], and e-waste contact history (Hg in cord blood, Cu, Zn, and Cd in maternal blood). CONCLUSIONS: Women in the e-waste recycling area had higher toxic element levels in the placenta and blood samples. More preventive measures were needed to reduce the risk of element exposure for mothers and fetuses in these areas.

A Questionnaire-Based Ensemble Learning Model to Predict the Diagnosis of Vertigo: Model Development and Validation Study.

BACKGROUND: Questionnaires have been used in the past 2 decades to predict the diagnosis of vertigo and assist clinical decision-making. A questionnaire-based machine learning model is expected to improve the efficiency of diagnosis of vestibular disorders. OBJECTIVE: This study aims to develop and validate a questionnaire-based machine learning model that predicts the diagnosis of vertigo. METHODS: In this multicenter prospective study, patients presenting with vertigo entered a consecutive cohort at their first visit to the ENT and vertigo clinics of 7 tertiary referral centers from August 2019 to March 2021, with a follow-up period of 2 months. All participants completed a diagnostic questionnaire after eligibility screening. Patients who received only 1 final diagnosis by their treating specialists for their primary complaint were included in model development and validation. The data of patients enrolled before February 1, 2021 were used for modeling and cross-validation, while patients enrolled afterward entered external validation. RESULTS: A total of 1693 patients were enrolled, with a response rate of 96.2% (1693/1760). The median age was 51 (IQR 38-61) years, with 991 (58.5%) females; 1041 (61.5%) patients received the final diagnosis during the study period. Among them, 928 (54.8%) patients were included in model development and validation, and 113 (6.7%) patients who enrolled later were used as a test set for external validation. They were classified into 5 diagnostic categories. We compared 9 candidate machine learning methods, and the recalibrated model of light gradient boosting machine achieved the best performance, with an area under the curve of 0.937 (95% CI 0.917-0.962) in cross-validation and 0.954 (95% CI 0.944-0.967) in external validation. CONCLUSIONS: The questionnaire-based light gradient boosting machine was able to predict common vestibular disorders and assist decision-making in ENT and vertigo clinics. Further studies with a larger sample size and the participation of neurologists will help assess the generalization and robustness of this machine learning method.

ASB7 Is a Novel Regulator of Cytoskeletal Organization During Oocyte Maturation.

Ankyrin repeat and SOCS box (ASB) family members have a C-terminal SOCS box and an N-terminal ankyrin-related sequence of variable repeats. To date, the roles of ASB family members remain largely unknown. In the present study, by employing knockdown analysis, we investigated the effects of ASB7 on mouse oocyte meiosis. We show that specific depletion of ASB7 disrupts maturational progression and meiotic apparatus. In particular, abnormal spindle, misaligned chromosomes, and loss of cortical actin cap are frequently observed in ASB7-abated oocytes. Consistent with this observation, incidence of aneuploidy is increased in these oocytes. Meanwhile, confocal scanning reveals that loss of ASB7 impairs kinetochore-microtubule interaction and provokes the spindle assembly checkpoint during oocyte meiosis. Furthermore, we find a significant reduction of ASB7 protein in oocytes from aged mice. Importantly, increasing ASB7 expression is capable of partially rescuing the maternal age-induced meiotic defects in oocytes. Together, our data identify ASB7 as a novel player in regulating cytoskeletal organization and discover the potential effects of ASB7 on quality control of aging oocytes.

The Regenerative Potential of Facial Nerve Motoneurons following Chronic Axotomy in Rats.

Background: The precise mechanisms of nerve regeneration remain unclear. The potential of facial nerve regeneration and probable mechanisms involved following chronic facial nerve injury should be further studied. Methods: Adult male Wistar rats were used to model either (i) facial nerve injury (axotomy) or (ii) reinjury (chronic axotomy followed by a second axotomy within 5 months). The rats were housed in the animal facility of the Eye and ENT Hospital of Shanghai Medical School, Fudan University (Shanghai, China). Expression of Shh (sonic hedgehog) and growth-associated protein 43 (GAP43, a neuronal marker) was detected in bilateral facial nuclei using reverse transcriptase PCR, western blotting analysis, and immunohistochemistry. The number of surviving motoneurons was quantified, and facial nerve regeneration was examined using transmission electron microscopy. Results: Reinjury of the facial nerve 12 weeks after the first axotomy resulted in upregulation of GAP43 mRNA and protein expression in neurons ipsilateral to the axotomy; immunohistochemistry revealed that Shh expression was higher compared with control side facial nuclei at the same time point. GAP43 expression subsequently decreased. Conclusion: The greatest regeneration potential of the facial nerve occurred within 5 months following chronic axotomy in rats, and regeneration may involve the Shh signaling pathway.

Molecular epidemiology of norovirus associated with acute gastroenteritis in Taizhou, China: A retrospective study.

BACKGROUND: Norovirus (NoV) is recognized as a leading global cause of viral acute gastroenteritis (AGE). To better understand the prevalence and genotypic patterns of NoV infection in Southeast China, we conducted a retrospective study of diarrheal syndrome surveillance of NoV and analyzed the epidemiological characteristics of AGE cases and phylogenetic evolution of NoV strains. METHODS: 1464 AGE patients in two diarrhea surveillance sentinel hospitals were sampled during 2016 and 2017. NoV Positive samples were genotyped by ORF1/ORF2 overlapping regional gene sequencing. Sequences analyses of the NoV genotypes were confirmed by online NoV Genotyping Tool and the phylogenetic analysis was constructed by MEGA 7.0. RESULTS: 139 (9.49%) of the AGE specimens were NoV positive. The GII strain was the main geno-group in NoV infected patients. At least 12 NoV genotypes and seven recombinant strains were detected. Major NoV genotypic transformations were GII.Pe/GII.4, GII.P17/GII.17 and GII.Pe/GII.17 in 2016 to GII.P16/GII.2, GII.P17/GII.17 and GII.Pe/GII.4 in 2017. Phylogenetic analysis showed that GII.P16/GII.2 recombinant strains clustered with those detected in the USA, Russia and Japan in 2016. CONCLUSION: We characterized the molecular epidemiology of NoV infection in AGE patients during 2016-2017. The main three NoV GII genotypes circulating in the population of Taizhou were GII.P17/GII.17, GII.Pe/GII.4 and GII.P16/GII.2.The GII.P16/GII.2 genotype has become the predominant strain since first quarter 2017. Monitoring of the NoV genotypic shift is important for the prevention and control of AGE.

Rab24 functions in meiotic apparatus assembly and maturational progression in mouse oocyte.

Rab GTPases have multiple regulatory functions in intracellular vesicle transport. In recent years, there has been an increasing interest in the roles of Rab proteins in mammalian oocytes. In this paper, we show the specific distribution pattern of Rab24 during mouse oocyte meiosis. Furthermore, we find that Rab24 depletion results in the failure of maturational progression in mouse oocytes. Notably, the frequency of meiotic apparatus abnormality is significantly increased in Rab24-depleted oocytes relative to controls. In addition, lagging chromosomes are readily observed in anaphase/telophase oocytes with Rab24 knockdown. In support of this, the depletion of Rab24 disturbs the kinetochore-microtubule attachments in oocytes, and contributes to the production of aneuploid eggs. Taken together, the results of this study identify Rab24 as a novel factor in the modulation of meiotic apparatus assembly and meiotic progression during mouse oocyte maturation.

NMNAT2-mediated NAD+ generation is essential for quality control of aged oocytes.

Advanced maternal age has been reported to impair oocyte quality; however, the underlying mechanisms remain to be explored. In the present study, we identified the lowered NAD+ content and decreased expression of NMNAT2 protein in oocytes from old mice. Specific depletion of NMNAT2 in mouse oocytes disturbs the meiotic apparatus assembly and metabolic activity. Of note, nicotinic acid supplementation during in vitro culture or forced expression of NMNAT2 in aged oocytes was capable of reducing the reactive oxygen species (ROS) production and incidence of spindle/chromosome defects. Moreover, we revealed that activation or overexpression of SIRT1 not only partly prevents the deficient phenotypes of aged oocytes but also ameliorates the meiotic anomalies and oxidative stress in NMNAT2-depleted oocytes. To sum up, our data indicate a role for NMNAT2 in controlling redox homeostasis during oocyte maturation and uncover that NMNAT2- NAD+ -SIRT1 is an important pathway mediating the effects of maternal age on oocyte developmental competence.

Mitofusin1 in oocyte is essential for female fertility.

Mitofusins (Mfn) are the important regulators of mitochondrial organization in mammalian cells; however, their roles during oocyte development remain unknown. In the present study, we generated mice with oocyte-specific knockout of Mfn1 or Mfn2 (Mfn1fl/fl;Zp3-Cre or Mfn2fl/fl;Zp3-Cre). We report that deletion of Mfn1, but not Mfn2, in oocytes leads to female mice sterility, associated with the defective folliculogenesis and impaired oocyte quality. In specific, follicles are arrested at secondary stage in Mfn1fl/fl;Zp3-Cre mice, accompanying with the reduced proliferation of granulosa cells. Moreover, alterations of mitochondrial structure and distribution pattern are readily observed in Mfn1-null oocytes. Consistent with this, mitochondrial activity and function are severely disrupted in oocytes from Mfn1fl/fl;Zp3-Cre mice. In addition, the differentially expressed genes in Mfn1-deleted oocytes are also identified by whole-transcriptome sequencing. In sum, these results demonstrate that Mfn1-modulated mitochondrial function is essential for oocyte development and folliculogenesis, providing a novel mechanism determining female fertility.

Histone methyltransferase SETD2 is required for meiotic maturation in mouse oocyte.

SET-domain-containing 2 (SETD2), a member of the histone lysine methyltransferase family, has been reported to be involved in multiple biological processes. However, the function of SETD2 during oocyte maturation has not been addressed. In this study, we find that mouse oocytes are incapable of progressing through meiosis completely once SETD2 is specifically depleted. These oocytes present an abnormal spindle morphology and deficient chromosome movement, with disrupted kinetochore-microtubule attachments, consequently producing aneuploidy eggs. In line with this, the BubR1 signal is markedly elevated in metaphase kinetochores of oocytes with SETD2 depletion, indicative of the activation of spindle assembly checkpoint. In addition, we note that loss of SETD2 results in a drastic decrease in the trimethylation level of H3K36 in oocytes. Collectively, our data demonstrate that SETD2 is required for oocyte maturation and indicate a novel mechanism controlling the meiotic apparatus.

SIRT7 functions in redox homeostasis and cytoskeletal organization during oocyte maturation.

SIRT7, a member of the sirtuin family, with coenzyme NAD catalyzes protein deacetylation and has been implicated in multiple biologic processes; however, its function in mammalian oocytes remains to be explored. Here, we report disrupted meiotic maturation upon specific knockdown of SIRT7 in mouse oocytes. In particular, disorganized spindle/chromosomes and the loss of the cortical actin cap are readily observed in SIRT7-depleted oocytes, generating aneuploid eggs. Furthermore, we found that SIRT7 depletion markedly elevated reactive oxygen species levels in oocytes, thereby compromising the developmental competence of early embryos. Of note, SIRT7 protein level is significantly decreased in oocytes from obese mice, and the forced expression of exogenous SIRT7 ameliorates maternal obesity-associated meiotic defects and oxidative stress in oocytes. In summary, our data suggest that SIRT7 is an essential factor in the determination of oocyte quality and may mediate the effects of obesity on female reproduction.-Gao, M., Li, X., He, Y., Han, L., Qiu, D., Ling, L., Liu, H., Liu, J., Gu, L. SIRT7 functions in redox homeostasis and cytoskeletal organization during oocyte maturation.

SIRT4 is essential for metabolic control and meiotic structure during mouse oocyte maturation.

SIRT4 modulates energy homeostasis in multiple cell types and tissues. However, its role in meiotic oocytes remains unknown. Here, we report that mouse oocytes overexpressing SIRT4 are unable to completely progress through meiosis, showing the inadequate mitochondrial redistribution, lowered ATP content, elevated reactive oxygen species (ROS) level, with the severely disrupted spindle/chromosome organization. Moreover, we find that phosphorylation of Ser293-PDHE1α mediates the effects of SIRT4 overexpression on metabolic activity and meiotic events in oocytes by performing functional rescue experiments. By chance, we discover the SIRT4 upregulation in oocytes from aged mice; and importantly, the maternal age-associated deficient phenotypes in oocytes can be partly rescued through the knockdown of SIRT4. These findings reveal the critical role for SIRT4 in the control of energy metabolism and meiotic apparatus during oocyte maturation and indicate that SIRT4 is an essential factor determining oocyte quality.

Sirt2-BubR1 acetylation pathway mediates the effects of advanced maternal age on oocyte quality.

The level of Sirt2 protein is reduced in oocytes from aged mice, while exogenous expression of Sirt2 could ameliorate the maternal age-associated meiotic defects. To date, the underlying mechanism remains unclear. Here, we confirmed that specific depletion of Sirt2 disrupts maturational progression and spindle/chromosome organization in mouse oocytes, with compromised kinetochore-microtubule attachments. Candidate screening revealed that acetylation state of lysine 243 on BubR1 (BubR1-K243, an integral part of the spindle assembly checkpoint complex) functions during oocyte meiosis, and acetylation-mimetic mutant BubR1-K243Q results in the very similar phenotypes as Sirt2-knockdown oocytes. Furthermore, we found that nonacetylatable-mimetic mutant BubR1-K243R partly prevents the meiotic deficits in oocytes depleted of Sirt2. Importantly, BubR1-K243R overexpression in oocytes derived from aged mice markedly suppresses spindle/chromosome anomalies and thereupon lowers the incidence of aneuploid eggs. In sum, our data suggest that Sirt2-dependent BubR1 deacetylation involves in the regulation of meiotic apparatus in normal oocytes and mediates the effects of advanced maternal age on oocyte quality.

HDAC3 promotes meiotic apparatus assembly in mouse oocytes by modulating tubulin acetylation.

Histone deacetylases (HDACs) have been shown to deacetylate numerous cellular substrates that govern a wide array of biological processes. HDAC3, a member of the Class I HDACs, is a highly conserved and ubiquitously expressed protein. However, its roles in meiotic oocytes are not known. In the present study, we find that mouse oocytes depleted of HDAC3 are unable to completely progress through meiosis, and are blocked at metaphase I. These HDAC3 knockdown oocytes show spindle/chromosome organization failure, with severely impaired kinetochore-microtubule attachments. Consistent with this, the level of BubR1, a central component of the spindle assembly checkpoint, at kinetochores is dramatically increased in metaphase oocytes following HDAC3 depletion. Knockdown and overexpression experiments reveal that HDAC3 modulates the acetylation status of α-tubulin in mouse oocytes. Importantly, the deacetylation mimetic mutant tubulin-K40R can partly rescue the defective phenotypes of HDAC3 knockdown oocytes. Our data support a model whereby HDAC3, through deacetylating tubulin, promotes microtubule stability and the establishment of kinetochore-microtubule interaction, consequently ensuring proper spindle morphology, accurate chromosome movement and orderly meiotic progression during oocyte maturation.

Sirt3-dependent deacetylation of SOD2 plays a protective role against oxidative stress in oocytes from diabetic mice.

Maternal diabetes has been demonstrated to adversely affect oocyte quality in mouse oocytes. However, the potential molecular mechanisms are poorly understood. Here, we established a type I diabetic mouse model and detected the increased reactive oxygen species (ROS) levels and decreased Sirt3 expression in oocytes from diabetic mice. Furthermore, we found that forced expression of Sirt3 in diabetic oocytes significantly attenuates such an excessive production of ROS. The acetylation status of lysine 68 of superoxide dismutase (SOD2K68) is dependent on Sirt3 in oocytes. In line with this, SOD2K68 acetylation levels were markedly increased in diabetic oocytes, and Sirt3 overexpression could effectively suppress this tendency. Importantly, the deacetylation-mimetic mutant SOD2K68R is capable of partly preventing the oxidative stress in oocytes from diabetic mice. In conclusion, our findings support a model where Sirt3 plays a protective role against oxidative stress in oocytes exposed to maternal diabetes through deacetylating SOD2K68.

Coxsackievirus A16 induced neurological disorders in young gerbils which could serve as a new animal model for vaccine evaluation.

Coxsackievirus A16 (CA16) is one of the major pathogens associated with human hand, foot, and mouth disease (HFMD) in the Asia-pacific region. Although CA16 infections are generally mild, severe neurological manifestations or even death has been reported. Studies on CA16 pathogenesis and vaccine development are severely hampered because the small animal models that are currently available show major limitations. In this study, gerbils (Meriones unguiculatus) were investigated for their suitability as an animal model to study CA16 pathogenesis and vaccine development. Our results showed that gerbils up to the age of 21 days were fully susceptible to CA16 and all died within five days post-infection. CA16 showed a tropism towards the skeletal muscle, spinal cord and brainstem of gerbils, and severe lesions, including necrosis, were observed. In addition, an inactivated CA16 whole-virus vaccine administrated to gerbils was able to provide full protection to the gerbils against lethal doses of CA16 strains. These results demonstrate that gerbils are a suitable animal model to study CA16 infection and vaccine development.

HIV related risky behavioral network and molecular epidemiology of HIV seroconcordant couples in Taizhou municipality of Zhejiang Province / 浙江大学学报·医学版

<p><b>UNLABELLED</b>Obstract: To characterize and analyze risky sexual networks and genetic scales to potential HIV transmission for HIV seroconcordant couples in Taizhou municipality of Zhejiang Province.</p><p><b>METHODS</b>HIV seroconcordant positive couples were invited as index cases to participate in an egocentric survey on HIV related risky behavior and behavioral network prior to HIV diagnosis during 2008-2011. Within-couple HIV transmission pairs were determined by the combination of both behavioral and phylogenetic analysis.</p><p><b>RESULTS</b>Totally 27 HIV seroconcordant couples were enrolled in this study. Male spouses were more likely to report having two or more sexual partners in the past years prior to HIV diagnosis than female spouses (88.9% vs. 37.0%). Among 27 couples, 20 couples including 17 couples by male but not female spouses, 3 couples by female but not male spouses reported having two or more sexual partners (i.e., multiple sexual partners) prior to HIV diagnosis; and 7 couples by both spouses reported having multiple sexual partners. Twenty four of 27 sexual networks were determined to be HIV transmission pairs (20) or potential transmission pairs (4), 3 couples were subtyped with discordant HIV subtypes or large genetic distance and thus had different sources of HIV transmissions. In addition, among 27 concordant couples, HIV drug resistance (HIVDR) or primary HIVDR existed in 6 ART-naïve participants in 4 networks; among them, 2 networks were determined to be potential HIVDR transmission couple pairs.</p><p><b>CONCLUSIONS</b>The HIV strains isolated in HIV infected spouses characterized with diversity and CRF01_AE was the main strain subtype. One of the spouses with risky behavior infected HIV was the main route of transmission to other spouses through unprotected sexual contacts. HIVDR was isolated from some HIV infected individuals, suggesting the risk for HIVDR transmission in married couples. The results provide enhanced evidence for urgent development of tailored prevention strategies, such as couple-based HIV counseling and testing services to reduce HIV secondary transmission.</p>

[A case of acute myelocytic leukemia suffered throat pain as initial symptom].

We report a case of a 47-year-old man who firstly complained of throat pain for half a month accompanied with fever. Specialized examination showed tonsils' hypertrophy and the laryngoscope found his right vocal cord was swelling and hyperemia. The routine blood test counted white blood cell as 31 x 10(9)/L, lymphocyte as 30. 84 x 10(9)/L while prolymphocyte could be seen with microscope. After that B-ultrasound scan gave spleen hypertrophy and multi-lymphatic-node enlargement. Peripheral blood flowcytometry and bone marrow biopsy finally diagnosed the man as actue myelocytic leukemia.

Behavioral and molecular tracing of risky sexual contacts in a sample of Chinese HIV-infected men who have sex with men.

Contact tracing, coupled with molecular epidemiologic investigation, is especially useful for identifying an infection with few cases in the population, such as human immunodeficiency virus (HIV) infection in China. No such research is available on Chinese men who have sex with men (MSM). From 2008 to 2010 in Taizhou Prefecture in China, every newly diagnosed HIV-infected MSM was invited to participate as an "index case" in a contact tracing survey by providing contact information for up to 8 sexual contacts, who themselves were approached to receive voluntary HIV counseling and testing. Those who tested HIV-positive were then subjected to another contact tracing survey. This process was repeated until no more sexual contacts were reported or tested positive. A total of 100 HIV-infected MSM served as "index cases," including the initial 49 cases identified through routine surveillance programs and 51 cases from the present survey. Traced MSM exhibited little willingness to receive voluntary counseling and testing. CRF01_AE (HIV type 1) was the dominant subtype. Seven of 49 independent sexual networks were deemed HIV transmission clusters. Fear of stigma or discrimination may deter Chinese MSM from receiving voluntary counseling and testing. Nonetheless, the integration of behavioral network analysis and HIV phylogenetic analysis provides enhanced evidence for developing tailored prevention strategies for HIV-infected MSM.

Tracing sexual contacts of HIV-infected individuals in a rural prefecture, Eastern China.

BACKGROUND: Contact tracing is especially useful for identifying an infection with few cases in the population, such as HIV in China. Little such research is available in China. METHODS: Every newly diagnosed HIV case from 2008-2010 in Taizhou Prefecture, Zhejiang Province in China, was invited to participate as an "index case" in a contact tracing survey by providing contact information for up to eight sexual contacts who themselves were approached for voluntary HIV counseling and testing (VCT). Those who tested HIV-positive were then subjected to another contact tracing survey. This process was repeated until no more sexual contacts were reported or tested positive. RESULTS: A total of 463 HIV-infected individuals were newly identified during the study period, including 338 cases who were identified from routine surveillance programs and 125 cases who were identified from the present contact tracing survey. Among these 463 cases, 398 (86.0%) served as 'index cases' in the survey, including 290 (85.8%) out of the 338 cases identified from routine surveillance programs and 108 (86.4%) out of the 125 cases identified from the present survey. These 398 'index cases' reported a total of 1,403 contactable sexual contacts, of whom 320 (22.8%) received HIV testing and 125 (39.1%) tested positive for HIV. Willingness to receive HIV testing was high among spouses and long term heterosexual or homosexual partners but extremely low among casual and commercial sex partners of 'index cases'. Consistent condom use was rare for all participants. A total of 290 independent sexual network components were constructed, with high complexity. CONCLUSION: Contact tracing is useful for identifying new HIV infections from spouses or long term sexual partners of HIV-infected individuals. The complicated sexual networks existing between and beyond HIV-infected persons provide opportunities for rapid spread of HIV in such areas.