Functions of FACT in Breaking the Nucleosome and Maintaining Its Integrity at the Single-Nucleosome Level.

The human FACT (facilitates chromatin transcription) complex, composed of two subunits SPT16 (Suppressor of Ty 16) and SSRP1 (Structure-specific recognition protein-1), plays essential roles in nucleosome remodeling. However, the molecular mechanism of FACT reorganizing the nucleosome still remains elusive. In this study, we demonstrate that FACT displays dual functions in destabilizing the nucleosome and maintaining the original histones and nucleosome integrity at the single-nucleosome level. We found that the subunit SSRP1 is responsible for maintenance of nucleosome integrity by holding the H3/H4 tetramer on DNA and promoting the deposition of the H2A/H2B dimer onto the nucleosome. In contrast, the large subunit SPT16 destabilizes the nucleosome structure by displacing the H2A/H2B dimers. Our findings provide mechanistic insights by which the two subunits of FACT coordinate with each other to fulfill its functions and suggest that FACT may play essential roles in preserving the original histones with epigenetic identity during transcription or DNA replication.

H2A mono-ubiquitination differentiates FACT's functions in nucleosome assembly and disassembly.

The histone chaperone FACT (FAcilitates Chromatin Transcription) plays an essential role in transcription and DNA replication by its dual functions on nucleosome assembly to maintain chromatin integrity and nucleosome disassembly to destabilize nucleosome and facilitate its accessibility simultaneously. Mono-ubiquitination at Lysine 119 of H2A (ubH2A) has been suggested to repress transcription by preventing the recruitment of FACT at early elongation process. However, up to date, how ubH2A directly affects FACT on nucleosome assembly and disassembly remains elusive. In this study, we demonstrated that the dual functions of FACT are differently regulated by ubH2A. The H2A ubiquitination does not affect FACT's chaperone function in nucleosome assembly and FACT can deposit ubH2A-H2B dimer on tetrasome to form intact nucleosome. However, ubH2A greatly restricts FACT binding on nucleosome and inhibits its activity of nucleosome disassembly. Interestingly, deubiquitination of ubH2A rescues the nucleosome disassembly function of FACT to activate gene transcription. Our findings provide mechanistic insights of how H2A ubiquitination affects FACT in breaking nucleosome and maintaining its integrity, which sheds light on the biological function of ubH2A and various FACT's activity under different chromatin states.

Effectiveness of enhancing contact model on reducing family caregiving burden and improving psychological wellbeing among caregivers of persons with schizophrenia in rural China.

BACKGROUND: It is unclear whether the enhancing contact model (ECM) intervention is effective in reducing family caregiving burden and improving hope and quality of life (QOL) among family caregivers of persons with schizophrenia (FCPWS). METHODS: We conducted a cluster randomized controlled trial in FCPWS in eight rural townships in Xinjin, Chengdu, China. In total, 253 FCPWS were randomly allocated to the ECM, psychoeducational family intervention (PFI), or treatment as usual (TAU) group. FCPWS in three groups were assessed caregiving burden, QOL and state of hope at baseline (T0), post-intervention (T1), 3-month (T2), and 9-month (T3) follow-up, respectively. RESULTS: Compared with participants in the TAU group, participants in the ECM group had statistically significantly lower caregiving burden scores both at T1 and T2 (p = 0.0059 and 0.0257, respectively). Compared with participants in the TAU group, participants in the PFI group had statistically significantly higher QOL scores in T1 (p = 0.0406), while participants in the ECM group had statistically significantly higher QOL scores in T3 (p = 0.0240). Participants in both ECM and PFI groups had statistically significantly higher hope scores than those in the TAU group at T1 (p = 0.0160 and 0.0486, respectively). CONCLUSIONS: This is the first study to explore the effectiveness of ECM on reducing family caregiving burden and improving hope and QOL in rural China. The results indicate the ECM intervention, a comprehensive and multifaceted intervention, is more effective than the PFI in various aspects of mental wellbeing among FCPWS. Future research needs to confirm ECM's effectiveness in various population.

Quality of life among family caregivers of people with schizophrenia in rural China.

PURPOSES: To investigate quality of life (QoL) of family caregivers of people with schizophrenia and examine the influencing factors of the QoL in a Chinese rural area. METHODS: This study included people with schizophrenia (n = 269) and their family caregivers (n = 269) from Xinjin district, Chengdu, China. Family caregivers' QoL was measured by the World Health Organization Quality of Life-Brief Form and its influencing factors was analyzed by the multivariate regression. RESULTS: Family caregivers of people with schizophrenia had very poor QoL across four domains. The regression analysis showed that physical domain of QoL was significantly associated with age, psychiatric symptoms of people with schizophrenia, and caregiving burden of family caregivers (p < 0.05). Psychological domain of QoL was significantly related to family caregivers' affiliate stigma, caregiving burden, and psychiatric symptoms of people with schizophrenia (p < 0.05). Social domain of QoL was significantly associated with age and psychiatric symptoms of people with schizophrenia, and affiliate stigma of family caregivers (p < 0.05). Environmental domain of QoL was significantly related to age and psychiatric symptoms of people with schizophrenia, and family caregivers' caregiving burden (p < 0.05). CONCLUSION: Family caregivers of people with schizophrenia had poor QoL in rural China. Family caregivers' QoL is significantly impacted by age and psychiatric symptoms of people with schizophrenia, caregivers' affiliate stigma and caregiving burden. Providing social support and psychosocial interventions for family caregivers of people with schizophrenia might be crucial to improve their QoL and caregiving.

Medication non-adherence and its influencing factors in persons with schizophrenia in rural China.

BACKGROUND: Few studies have been conducted to explore medication non-adherence in persons with schizophrenia (PWS) and its influencing factors in rural China. This study aimed to investigate the medication non-adherence and its influencing factors among PWS in rural China. METHODS: A total of 269 PWS and their family caregivers in Xinjin district, Chengdu, China were investigated on medication adherence and related factors. Logistic regression was employed to identify the influencing factors. RESULTS: The results showed that 37.6% of PWS had medication non-adherence. PWS living with family caregivers had significantly lower rate of medication non-adherence (34.7%) than those not living with family caregivers (60.0%) (p < 0.01). Family caregivers' affiliate stigma of mental illness, knowledge of mental illness and self-esteem were significantly related to patients' medication non-adherence (p < 0.05). PWS' employment status, living with family caregiver, present mental status and social support were significantly related to medication adherence. CONCLUSION: This study shows medication non-adherence is severe among PWS in rural China. Both patient- and family-related factors affect patients' medication adherence seriously. Except improving patients' treatment and mental status, development of family caregiving, social support network and intervention on reducing stigma of mental illness should be crucial for enhancing PWS' medication adherence.

Curaxin-Induced DNA Topology Alterations Trigger the Distinct Binding Response of CTCF and FACT at the Single-Molecule Level.

The candidate anticancer drug curaxins can insert into DNA base pairs and efficiently inhibit the growth of various cancers. However, how curaxins alter the genomic DNA structure and affect the DNA binding property of key proteins remains to be clarified. Here, we first showed that curaxin CBL0137 strongly stabilizes the interaction between the double strands of DNA and reduces DNA bending and twist rigidity simultaneously, by single-molecule magnetic tweezers. More importantly, we found that CBL0137 greatly impairs the binding of CTCF but facilitates trapping FACT on DNA. We revealed that CBL0137 clamps the DNA double helix that may induce a huge barrier for DNA unzipping during replication and transcription and causes the distinct binding response of CTCF and FACT on DNA. Our work provides a novel mechanical insight into CBL0137's anticancer mechanisms at the nucleic acid level.

Natural history of pathologically confirmed pulmonary subsolid nodules with deep learning-assisted nodule segmentation.

OBJECTIVE: To explore the natural history of pulmonary subsolid nodules (SSNs) with different pathological types by deep learning-assisted nodule segmentation. METHODS: Between June 2012 and June 2019, 95 resected SSNs with preoperative long-term follow-up were enrolled in this retrospective study. SSN detection and segmentation were performed on preoperative follow-up CTs using the deep learning-based Dr. Wise system. SSNs were categorized into invasive adenocarcinoma (IAC, n = 47) and non-IAC (n = 48) groups; according to the interval change during the preoperative follow-up, SSNs were divided into growth (n = 68), nongrowth (n = 22), and new emergence (n = 5) groups. We analyzed the cumulative percentages and pattern of SSN growth and identified significant factors for IAC diagnosis and SSN growth. RESULTS: The mean preoperative follow-up was 42.1 ± 17.0 months. More SSNs showed growth or new emergence in the IAC than in the non-IAC group (89.4% vs. 64.6%, p = 0.009). Volume doubling time was non-significantly shorter for IACs than for non-IACs (1436.0 ± 1188.2 vs. 2087.5 ± 1799.7 days, p = 0.077). Median mass doubling time was significantly shorter for IACs than for non-IACs (821.7 vs. 1944.1 days, p = 0.001). Lobulated sign (p = 0.002) and SSN mass (p = 0.004) were significant factors for differentiating IACs. IACs showed significantly higher cumulative growth percentages than non-IACs in the first 70 months of follow-up. The growth pattern of SSNs may conform to the exponential model. The initial volume (p = 0.042) was a predictor for SSN growth. CONCLUSIONS: IACs appearing as SSNs showed an indolent course. The mean growth rate was larger for IACs than for non-IACs. SSNs with larger initial volume are more likely to grow. KEY POINTS: • Invasive adenocarcinomas (IACs) appearing as subsolid nodules (SSNs), with a mean volume doubling time (VDT) of 1436.0 ± 1188.2 days and median mass doubling time (MDT) of 821.7 days, showed an indolent course. • The VDT was shorter for IACs than for non-IACs (1436.0 ± 1188.2 vs. 2087.5 ± 1799.7 days), but the difference was not significant (p = 0.077). The median MDT was significantly shorter for IACs than for non-IACs (821.7 vs. 1944.1 days, p = 0.001). • SSNs with lobulated sign and larger mass (> 390.5 mg) may very likely be IACs. SSNs with larger initial volume are more likely to grow.

Analysis of Chemical Components in the Roots of Eight Intersubgeneric Hybrids of Paeonia.

Paeonia cultivars are famous ornamental plants, and some of them are also traditional Chinese medicinal resources. Intersubgeneric hybrids of Paeonia (IHPs) are formed by the hybridization of herbaceous peony (Paeonia lactiflora) and tree peony (Paeonia×suffruticosa or lutea hybrid tree peony). The phenotypic characteristics of IHPs are similar to those of herbaceous peony, and their root systems are large and vigorous. However, their medicinal value has not been reported yet. In this study, the roots of eight IHP samples were analyzed by high performance liquid chromatography quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS/MS). A total of 18 compounds were identified, including phenols, paeonols, monoterpene glycosides, and tannins. The contents of monoterpene glycosides and tannins in IHPs were higher than herbaceous peony and tree peony, exceeding 44.76 mg/g DW and 11.50 mg/g DW, respectively. Three IHPs, 'Prairie Charm', 'Garden Treasure', and 'Yellow Emperor', with more types and a higher content of medicinal compounds, were screened out by cluster analysis. These IHPs have considerable potential for the development of medicinal resources.

Histone H2A Ubiquitination Reinforces Mechanical Stability and Asymmetry at the Single-Nucleosome Level.

Monoubiquitination at lysine 119 of histone H2A (ubH2A) is a prevalent post-translational modification that is associated with gene repression in the context of chromatin. However, the direct function of ubH2A on nucleosome is poorly understood. Here we identified the effect of ubH2A on nucleosome using single-molecule magnetic tweezers. We revealed that ubH2A stabilizes the nucleosome by blocking the peeling of DNA from the histone octamer. Each ubH2A reinforces one-half of the outer wrap and introduces a robust asymmetry for nucleosome unfolding. Furthermore, a real-time deubiquitination process confirmed that ubH2A-nucleosome is sequentially deubiquitinated and restored to the unmodified nucleosome state. These results provide a novel mechanism to understand the repression of the passage of RNA or DNA polymerases through the ubH2A-nucleosome barrier during gene transcription or replication.

Long-term follow-up of persistent pulmonary pure ground-glass nodules with deep learning-assisted nodule segmentation.

OBJECTIVE: To investigate the natural history of persistent pulmonary pure ground-glass nodules (pGGNs) with deep learning-assisted nodule segmentation. METHODS: Between January 2007 and October 2018, 110 pGGNs from 110 patients with 573 follow-up CT scans were included in this retrospective study. pGGN automatic segmentation was performed on initial and all follow-up CT scans using the Dr. Wise system based on convolution neural networks. Subsequently, pGGN diameter, density, volume, mass, volume doubling time (VDT), and mass doubling time (MDT) were calculated automatically. Enrolled pGGNs were categorized into growth, 52 (47.3%), and non-growth, 58 (52.7%), groups according to volume growth. Kaplan-Meier analyses with the log-rank test and Cox proportional hazards regression analysis were conducted to analyze the cumulative percentages of pGGN growth and identify risk factors for growth. RESULTS: The mean follow-up period of the enrolled pGGNs was 48.7 ± 23.8 months. The median VDT of the 52 pGGNs having grown was 1448 (range, 339-8640) days, and their median MDT was 1332 (range, 290-38,912) days. The 12-month, 24.7-month, and 60.8-month cumulative percentages of pGGN growth were 10%, 25.5%, and 51.1%, respectively, and they significantly differed among the initial diameter, volume, and mass subgroups (all p < 0.001). The growth pattern of pGGNs may conform to the exponential model. Lobulated sign (p = 0.044), initial mean diameter (p < 0.001), volume (p = 0.003), and mass (p = 0.023) predicted pGGN growth. CONCLUSIONS: Persistent pGGNs showed an indolent course. Deep learning can assist in accurately elucidating the natural history of pGGNs. pGGNs with lobulated sign and larger initial diameter, volume, and mass are more likely to grow. KEY POINTS: • The pure ground-glass nodule (pGGN) segmentation accuracy of the Dr. Wise system based on convolution neural networks (CNNs) was 96.5% (573/594). • The median volume doubling time (VDT) of 52 pure ground-glass nodules (pGGNs) having grown was 1448 days (range, 339-8640 days), and their median mass doubling time (MDT) was 1332 days (range, 290-38,912 days). The mean time to growth in volume was 854 ± 675 days (range, 116-2856 days). • The 12-month, 24.7-month, and 60.8-month cumulative percentages of pGGN growth were 10%, 25.5%, and 51.1%, respectively, and they significantly differed among the initial diameter, volume, and mass subgroups (all p values < 0.001). The growth pattern of pure ground-glass nodules may conform to exponential model.

[Qilin Pills promote testicular spermatogenesis in azoospermia mice].

OBJECTIVE: To investigate the effect of Qilin Pills (QP) in facilitating the recovery of spermatogenic function in azoospermia (AS) mice and to explore its mechanism of regulating testicular spermatogenesis. METHODS: Fifteen 4-week-old male mice were equally randomized into an AS model control, a low-dose QP and a high-dose QP group. The AS model was established in the mice by intraperitoneal injection of busulfan at 35 mg/kg. After modeling, the animals in the low- and high-dose QP groups were treated with Qilin Pills intragastrically at 2 000 and 8 000 mg/kg/d respectively while those in the model control group fed on a normal diet, all for 28 days. Then, all the mice were sacrificed for examination of the ultrastructures of the epididymis and testis by HE staining, detection of the specific markers of spermatogenic, Sertoli and Leydig cells by Western blot, and determination of the expressions of these markers in the testis tissue by immunofluorescence assay. RESULTS: The number of spermatogenic cells in the testis tissue was significantly decreased in the AS model controls, with no spermatozoa in most of the seminiferous tubules in the epididymis (Johnsen's score: 5.2 ± 0.5). In the high-dose QP group, spermatogenic cells were tightly arranged with distinct layers in the seminiferous tubules, with a large number of spermatozoa but no non-sperm cells in the lumens of the epididymis (Johnsen's score: 9.4 ± 0.6). The number of spermatogenic cells in the testis was increased in the low-dose QP group with some spermatozoa in the seminiferous tubules as compared with that in the model control, but lower than in the high-dose group (Johnsen's score: 7.6 ± 0.6). The Johnsen's score was significantly lower in the model control than in the high- and low-dose QP groups (P < 0.01), and higher in the high-dose than in the low-dose QP group (P < 0.05). The expressions of the specific markers of Sertoli cells SCF, BMP4, SYCP3, DMC1 and Ki67 were also remarkably lower in the model control than in the high- and low-dose QP groups (P < 0.01), and higher in the high-dose than in the low-dose QP group (P < 0.05 or P < 0.01). No statistically significant differences were observed among the three groups of mice in the markers of spermatogonial stem cells (SSC) and undifferentiated SSCs UCHL1, STRA8, NGN3 and PLZF3 (P > 0.05). The expressions of the spermatocyte markers DMC1 and SYCP3 were markedly lower in the model control than in the high- and low-dose QP groups (P < 0.05 or P < 0.01), and higher in the high-dose than in the low-dose QP group (P < 0.05 or P < 0.01). The Ki67 fluorescence signals were distributed in the spermatogonia, with a higher intensity in the model control than in the high- and low-dose QP groups. The acrosome marker PNA was found mainly in the seminiferous tubules, with abundant fluorescence signals in the high- and low-dose QP groups but no obvious dot signals in the model controls. CONCLUSIONS: Qilin Pills may contribute to the meiosis of spermatogonia and promote spermatogenesis by improving the function of Sertoli cells in the testis.

Bioethanol production from the dry powder of Jerusalem artichoke tubers by recombinant Saccharomyces cerevisiae in simultaneous saccharification and fermentation.

It has been found that recombinant Saccharomyces cerevisiae 6525 can produce high concentration of ethanol in one-step fermentation from the extract of Jerusalem artichoke tubers or inulin. However, the utilization rate of raw materials was low and the fermentation process was costly and complicated. Therefore, in this study, after the optimum processing conditions for ethanol production in fed-batch fermentation were determined in flask, the recombinant S. cerevisiae 6525 was first used to produce ethanol from the dry powder of Jerusalem artichoke tubers in 5-L agitating fermentor. After 72 h of fermentation, around 84.3 g/L ethanol was produced in the fermentation liquids, and the conversion efficiency of inulin-type sugars to ethanol was 0.453, or 88.6 % of the theoretical value of 0.511. This study showed high feasibility of bioethanol industrial production from the Jerusalem artichoke tubers and provided a basis for it in the future.

[The cytotoxic effect and the effect of permeability with OmpA like protein Loa22 from Leptospira interrogans serovar].

OBJECTIVE: To study the toxic effect and the change of permeability on human umbilical vein endothelia (HUVE) of the Loa22 protein from virulent serovar Lai. Leptaspira interrogans by expressing its protein. METHODS: In this study, the pGEX-Loa22 peptide prokaryotic recombinant plasmid of Leptospira interrogans serovar Lai preserved in our laboratory was used to express Loa22 fusion protein with GST lable. Then the target fusion protein was obtained by using affinity chromatography with the GST-Trap FF Column. The purified Loa22 fusion protein was detected by SDS-PAGE and confirmed by Western blot assay using the mouse anti-GST tag monoclonal anti-body. pGEX-Loa22 protein was administered to culture with human umbilical vein endothelial cells (HUVEC) to elucidate the cytotoxic role and the change of permeability of leptospiral outer membrane proteins. RESULTS: The recombiant plasmid with Loa22 mature peptide was expressed successfully and the protein was purfied. Significant higher level of apoptosis ratio, lower CCK-8 aborntion, and increasing permeability on HUVEC were observed after treated the HUVEC with the expressed fusion protein. CONCLUSION: The purified Loa22 fusion protein have obvious toxic effects on vascular endothelial cells, and also it can increase permeability of HUVEC.

Affiliate stigma and caregiving burden among family caregivers of persons with schizophrenia in rural China.

BACKGROUND: Although stigma and caregiving burden are important in relation to mental health recovery, few studies have been conducted on affiliate stigma and caregiving burden among family caregivers of persons with schizophrenia (FCPWS) in rural China. AIMS: This study aimed to examine the severity level of affiliate stigma and caregiving burden, and identify the correlates among FCPWS in rural China. METHODS: A mental health survey was conducted (N = 253 FCPWS) in Xinjin county, Sichuan province, China. Affiliate Self-Stigma Scale and Zarit Burden Interview Short Form were used. The regression analysis was performed to explore the correlates of stigma and burden. RESULTS: Most FCPWS reported experiencing high and severe level of affiliate stigma (78.66%) and caregiving burden (95.26%). Family caregivers who were middle aged, unemployed, with high caregiving burden and low quality of life (QoL), showed more severe affiliate stigma. Family caregivers who were female, older, with low income, high affiliate stigma and low QoL, experienced greater caregiving burden. CONCLUSIONS: The large majority of FCPWS in rural China experienced severe affiliate stigma, caregiving burden and poor QoL. It is crucial to develop culture-specific anti-stigma interventions to reduce caregivers' stigma and caregiving burden, and improve QoL. Specific risk factors of family caregivers' affiliate stigma and caregiving burden should be considered for development of health policy and community-based mental health services.

Temporal and Spatial Dynamics of Organ Water Content in Maize with Different Senescence Types.

Understanding the water status of specific organs can be helpful in evaluating the life activities and growth conditions of maize. To accurately judge organ growth conditions and thus design appropriate interventions, it is necessary to clarify the true water dynamics of each maize organ. Using multiple maize cultivars with different growth periods, spatio-temporal water dynamics were analyzed here in the leaves, stalks, and ear components. Leaf water content was found to gradually decrease from both the bottom and top of the plant to the middle, whereas stalk water content decreased sequentially from the top to the bottom. Each successively higher node from the bottom of the plant was associated with decreases of 0.99% and 1.27% water content in the leaves and stalks, respectively. The water dynamics in leaves and internodes showed three clear stages: the slow loss, rapid loss, and balance stage. A water content of 60% appeared to be an irreversible turning point for initiation of senescence. Using normalized growth period as a measure, each of the tested cultivars could be assigned into one of two types based on their water dynamics: stay-water or general type. General-type cultivars had a shorter duration with a high water content and a water loss rate approximately twice as high as that of the stay-water type. This may have been related to the leaf senescence characteristics. However, the stay-water trait did not interfere with water dynamics of the ear components. Therefore, it may not be robust to evaluate the kernel dehydration of maize according to leaf senescence conditions due to the weak correlation between kernel water content and leaf senescence characteristics.

Physical illness comorbidity and its influencing factors among persons with severe mental illness in Rural China.

BACKGROUND: Comorbid physical illness is a common cause of death in people with severe mental illness (SMI) worldwide. In rural China, the prevalence of physical illness comorbidity among persons with SMI remains unclear. This study aimed to examine non-communicable physical illness comorbidity and its risk factors among people with SMI in a rural area of China. METHODS: A mental health survey, using the International Classification of Disease (ICD-10), was conducted in six townships of Xinjin District, Chengdu, China in 2015. RESULTS: A total of 724 persons with SMI were included in this study, and 37.8% of them had at least one physical illness. The most common physical illnesses were hypertension (10.5%) and diabetes (5.8%). More physical comorbidity was reported among persons with affective disorders than persons with schizophrenia. Many participants (37.4%) had never received antipsychotic treatment, and of those, 51.6% reported having a physical illness comorbidity. Significant associations were found between physical illness comorbidity and participants' family economic status, family size, age at onset of mental disorder, treatment status, and symptom severity. CONCLUSIONS: Our findings indicate the need of an integration of medical and psychiatric care in primary care. It also suggests that poverty and having never received treatment for mental health problems negatively affect the health of persons with SMI, which deserve more attention. Researchers and policymakers can take these findings into account to develop health policies and improve the mental and physical health care in rural China.

Effectiveness of enhancing contact model on reducing stigma of mental illness among family caregivers of persons with schizophrenia in rural China: A cluster randomized controlled trial.

Background: Contact-based intervention has been documented and proved effective on reducing stigma of mental illness in high-income countries, but it is still unclear about the effectiveness of the contact-based intervention among family caregivers of persons with schizophrenia (FCPWS) in low- and middle-income countries including rural China. Methods: We conducted a cluster randomized controlled trial in FCPWS in eight rural townships in Xinjin district of Chengdu city in Southwest China. The FCPWS in these townships were randomly allocated to the Enhancing Contact Model (ECM), Psychoeducational Family Intervention (PFI), or Treatment as Usual (TAU) group. FCPWS in three groups were provided specific interventions and follow-ups. By using a mixed-effect model, our goal was to examine the differences in affiliate self-stigma scale (ASSS) scores among three groups with the data collected at baseline (T0), post-intervention (T1), 3-month (T2), and 9-month (T3) follow-up timepoints, respectively. This trial is registered with ChiCTR, number ChiCTR2000039133. Findings: In April 2019, 253 FCPWS from 8 townships were randomly assigned to receive either ECM (cluster=3, n=90), PFI (cluster=2, n=81), or TAU (cluster=3, n=82). Compared with participants in the TAU group, participants in the ECM group had statistically significantly lower ASSS scores at 9-month follow-up (estimated parameter [EP]= -5.51, 95% CI -10.27 to -0.74, p=0.02). There were no statistically significantly different ASSS scores at 9-month follow up between ECM and PFI groups. Compared with participants in the PFI group, younger (<60 years old), with higher monthly income and other caregiver (e.g., parent, sibling, child) participants in the ECM group had statistically significantly lower ASSS scores in the 3-month follow-up (EP = -5.66, 95% CI -10.13 to -1.19, p<0.01; EP = -7.82, 95% CI -11.87 to -3.78, p<0.001; EP = -6.79, 95% CI -10.69 to -2.90, p<0.001, respectively). Interpretation: This first trial in rural China shows that ECM intervention, a new anti-stigma intervention model, is a promising method for reducing affiliate stigma among FCPWS. The ECM intervention is more effective and stable than the PFI on reducing affiliate stigma among FCPWS. Further research needs to explore whether a long-term intervention could produce a more positive anti-stigma outcome trajectory. Funding: General Research Fund, University Grants Committee, Hong Kong SAR (GRF, Grant No. 17605618, 2018-2021, PI: Dr. M.S. Ran).

Affiliate stigma of mental illness in family caregivers of persons with mental illness in Hong Kong.

BACKGROUND: Affiliate stigma of family caregivers may severely influence family caregiving, early treatment, and recovery of people with mental illness (PMI), and it may be impacted by the knowledge of mental illness and contact with PMI. However, little is known about the correlation between affiliate stigma of family caregivers of PMI and contact in Hong Kong. AIMS: This study aimed to investigate affiliate stigma and its influencing factors among family caregivers of PMI in Hong Kong. METHODS: A total of 106 family caregivers in Hong Kong participated in the study. The measurements included affiliate stigma, contacts (quantity, quality, and level), knowledge, prejudice, and discriminatory behaviors. Multiple regression analyses were employed. RESULTS: The mean score of affiliate stigma scale (ASS) in family caregivers was 2.17 (SD = 0.65). In regression analyses, the results showed that contact quantity and contact level were significantly associated with higher affiliate stigma and its cognitive domain, while contact quality (e.g. positive contact) were significantly associated with lower affiliate stigma and its all domains among family caregivers. The positive relationship between prejudicial attitudes and affiliate stigma was also found. CONCLUSIONS: Affiliate stigma is severe among family caregivers of PMI in Hong Kong. The results of this study indicate that contact, especially positive contact, contributes to reducing affiliate stigma among family caregivers of PMI. The results of this study are important for development of health policy on reducing stigma in family caregivers of PMI. The effectiveness of Enhancing Contact Model (ECM) should be examined in future anti-stigma interventions.

Novel mutation in ODF2 causes multiple morphological abnormalities of the sperm flagella in an infertile male.

Numerous genes have been associated with multiple morphological abnormalities of the sperm flagella (MMAF), which cause severe asthenozoospermia and lead to male infertility, while the causes of approximately 50% of MMAF cases remain unclear. To reveal the genetic causes of MMAF in an infertile patient, whole-exome sequencing was performed to screen for pathogenic genes, and electron microscope was used to reveal the sperm flagellar ultrastructure. A novel heterozygous missense mutation in the outer dense fiber protein 2 (ODF2) gene was detected, which was inherited from the patient's mother and predicted to be potentially damaging. Transmission electron microscopy revealed that the outer dense fibers were defective in the patient's sperm tail, which was similar to that of the reported heterozygous Odf2 mutation mouse. Immunostaining of ODF2 showed severe ODF2 expression defects in the patient's sperm. Therefore, it was concluded that the heterozygous mutation in ODF2 caused MMAF in this case. To evaluate the possibility of assisted reproductive technology (ART) treatment for this patient, intracytoplasmic sperm injection (ICSI) was performed, with the help of a hypo-osmotic swelling test and laser-assisted immotile sperm selection (LAISS) for available sperm screening, and artificial oocyte activation with ionomycin was applied to improve the fertilization rate. Four ICSI cycles were performed, and live birth was achieved in the LAISS-applied cycle, suggesting that LAISS would be valuable in ART treatment for MMAF.

Cryopiece, a novel carrier with faster cooling rate, high recovery rate and retrieval rate, for individual sperm cryopreservation.

BACKGROUND: Cryopreservation of extremely few spermatozoa is still a major challenge for male fertility preservation. This study aims to evaluate the cooling rate, recovery rate, and retrieval rate, along with other parameters of spermatozoa that cryopreserved using Cryopiece, a novel carrier, for individual sperm cryopreservation. METHODS: Semen samples from 60 fertile donors were collected, and each semen sample was screened for motile sperm and mixed with cryoprotective agent (CPA), and then frozen using Cryopiece, micro-straw, and mini-straws. The cooling rate, retrieval rate, and recovery rate, morphology, DNA fragmentation index (DFI) and mitochondrial membrane potential (MMP), were compared among the un-frozen sperm and the sperm cryopreserved using these carriers. RESULTS: Cryopiece possessed the fastest cooling rate. After freeze-thaw, the average retrieval rate of sperm cryopreserved using Cryopiece was 96.25%, and the average recovery rate was 64.40%, which were higher than that of sperm cryopreserved using the other two carriers (71.42% and 54.30% for micro-straw, and 63.54% and 58.04% for mini-straw, respectively). There was no significant impact on DFI after sperm cryopreservation, and no significant difference in morphology between sperm cryopreserved using these carriers was observed. Though MMP of sperm changed significantly after cryopreservation, micro-straw maintained sperm MMP better than Cryopiece and mini-straw did, while no significant difference was observed in MMP between sperm cryopreserved using Cryopiece and mini-straw. CONCLUSIONS: Cryopiece produced satisfying retrieval and recovery rates in sperm cryopreservation and should be an ideal carrier for cryopreservation of small number of sperm.