High-fidelity CRISPR/Cas12a dual-crRNA screening reveals novel synergistic interactions in hepatocellular carcinoma.

 : CRISPR/Cas12a-based combinational screening has shown remarkable potential for identifying genetic interactions. Here, we describe an innovative method for combinational genetic screening with rapid construction of a dual-CRISPR RNA (crRNA) library using gene splicing through overlap extension PCR (SOE PCR) and the adoption of CeCas12a, which we previously identified with strict PAM recognition and low off-targeting to guarantee fidelity and efficiency. The custom-pooled SOE crRNA array (SOCA) library for double-knockout screening could be conveniently constructed in the laboratory for widespread use, and the CeCas12a-mediated high-fidelity screen displayed good performance even under a negative selection screen. By designing a SOCA dual-crRNA library that covered most of the kinase and metabolism-associated gene targets of FDA-approved drugs implicated in hepatocellular carcinoma (HCC) tumourigenesis, novel cross-talk between the two gene sets was negatively selected to inhibit HCC cell growth in vitro and in vivo and was validated using virtual double-knockdown screening based on TCGA databases. Thus, this rapid, efficient and high-fidelity double-knockout screening system is promising for systemically identifying potential genetic interactions between multiple gene sets or combinations of FDA- approved drugs for clinical translational medicine in the future.

Understanding the Role of a Silane-Coupling Agent in Bio-Based Polyurethane Nanocomposite-Coated Fertilizers.

Bio-based polyurethane (PU)-coated controlled release fertilizers are attracting a lot of attention; however, generally they have poor properties, so it is difficult for them to meet the agricultural needs. Herein, γ-aminopropyl triethoxy silane (KH550) was first used with nanosilica (NS) to prepare bio-based PU nanocomposite-coated urea (KSPCU). The coating microstructures and nutrient controlled release behaviors of KSPCU were investigated and compared with those of unmodified NS containing PU nanocomposite-coated urea (SPCU) and bio-based PU-coated urea (PCU). The KSPCU with KH550 exhibited an excellent controlled release performance. Its nutrient release longevity exceeded 105 d, which was nearly 6 times greater than that of PCU and 2 times more than that of SPCU, and it was much longer than that of PCU reported in previous research at a coating rate of 3 wt %. A series of characterization methods combined with water resistance capacity and porosity measurements confirmed that a hydrogen bond was formed by the reaction between the nanoparticle and PM200, the nanoparticle was bonded on the macromolecular chain, and KH550 in the coating increased the cross-linking degree, which were beneficial to slowing down the nutrient release of the KSPCU. The innovative application of KH550 on bio-based PU-coated fertilizers will provide a new coating technology for improving their controlled release property.

The Risk of Hypertension Doubles Every 10 Years in China: Age, Period, and Birth Cohort Effects on the Prevalence of Hypertension From 2004 to 2013.

BACKGROUND: Hypertension is a growing problem worldwide and can often result in a variety of negative health outcomes. The aim of this study was to assess the effects of age at diagnosis, calendar period, and birth cohort on the change in the prevalence rate of hypertension in Guangzhou from 2004 to 2013. METHODS: We used data from the Guangzhou Community Health Survey, a population-based study designed by the National Health and Family Planning Commission of the PRC every 5 years. A total of 27,299, 23,467, and 18,362 participants aged 15-79 years completed the survey in 2004, 2009, and 2013, respectively. RESULTS: Age effects increased slowly before the age of 42 years but increased rapidly after the age of 42 years, peaking at 79 years. Cohort effects grew slowly before the end of the 1960s but grew quickly after the end of the 1960s. The risk of suffering from hypertension among people born in 1962, 1972, 1982, and 1992 was 1.39, 2.68, 5.55, and 11.53 times, respectively, than that of people born in 1952. The period effects increased 25% from 2004 to 2009 and later declined 27% from 2009 to 2013 in the entire population. There was no gender difference in age effects and period effects, but strong cohort effects on hypertension were observed among males compared with females. CONCLUSIONS: For Chinese individuals, the later one is born, the higher the risk is of suffering from hypertension. Strong cohort effects for hypertension were observed among males compared with females, indicating that males are more easily affected by hypertension based on the change in birth cohort.

Cyberbullying perpetration and victimisation among junior and senior high school students in Guangzhou, China.

OBJECTIVES: Cyberbullying research in China is in early stage. This study describes the cyberbullying experiences of junior and senior high school students in Guangzhou, China, and to examine the risk factors associated with cyberbullying perpetrators, victims and perpetrator-victims among students. We also investigated the frequency of cyberbullying and coping strategies of student victims. METHODS: Participants were 2590 students in grades 7, 8, 9 and 10 from six junior and senior high schools in October 2015 in Guangzhou, in south China, who completed a questionnaire. Data on participants' experiences with cyberbullying perpetration and victimisation during the previous 6 months were collected. Multinomial logistic regression was used to analyse factors associated with being perpetrators, victims and perpetrator-victims. RESULTS: In this sample, 28.0% (725) of participants reported being a perpetrator and 44.5% (1150) reported being a victim in the previous 6 months. Specifically, 2.9% (74) reported being perpetrators only, 19.3% (499) reported being victims only and 25.2% (651) reported being perpetrator-victims (both perpetrator and victim). In addition, flaming was the most common form of cyberbullying in both perpetration and victimisation. Logistic regression analyses indicated that online game addiction in participants was associated with increased odds of being a perpetrator only; no democratic parenting style in the mother and physical discipline by parents were associated with increased odds of being a victim only; male students, students with low academic achievement, those spending over 2 hours a day online, experiencing physical discipline from parents and online game addiction were associated with increased odds of both perpetration and victimisation. CONCLUSIONS: Cyberbullying is a common experience among Chinese junior and senior high school students. These findings add to the empirical data on cyberbullying and reinforce the urgent need for cyberbullying prevention in China. Furthermore, from the perspective of practice, it is important to raise our awareness of cyerbullying and reduce the risk factors.

HIV-related stigma and discrimination amongst healthcare providers in Guangzhou, China.

BACKGROUND: HIV-related discrimination amongst healthcare providers is one of the strongest obstacles to effectively responding to HIV. This study was conducted to explore the occurrence of and other factors related to discrimination against people living with HIV/AIDS amongst healthcare providers in Guangzhou, China. METHODS: This was a cross-sectional study, conducted between July and October 2016, that enrolled healthcare providers from 9 healthcare institutions in Guangzhou, China. HIV-related discrimination was assessed using anonymous self-designed questionnaires. Chi-square tests were used to study the differences in the socio-demographic characteristics, occupational characteristics, HIV-related knowledge and personal attitudes between participants who had and had not discriminated against People living with HIV/AIDS (PLWHA). A multivariate logistic regression analysis was used to study the factors associated with HIV-related discrimination. RESULTS: A total of 972 healthcare providers were investigated, and 386 (39.7%) had previously served HIV-positive individuals in their work. Administering HIV antibody tests for patients without his or her consent was the most frequent act of discrimination (65.3%), and other forms of discrimination, including "differential treatment" (51.0%), "disclosed information" (46.4%) and "refused to treat" (38.6%), were also prevalent. The logistic regression analysis indicated that people who had worked for 3-7 years, worked in secondary hospitals or lower, worked in surgical departments, had lower scores on HIV transmission knowledge, were dissatisfied with the occupational exposure protection system offered by the government, were worried about HIV-related exposure and feared HIV-related exposure were more likely to commit an act of medical discrimination against PLWHA. CONCLUSION: HIV-related discrimination was not unusual in the healthcare providers of Guangzhou, which may be related to their negative cognitions and attitudes as well as the hospital management system and government policy. Therefore, comprehensive HIV-related knowledge education should be implemented to change the attitude of healthcare providers. In addition, the current laws and regulations should be refined by the government to protect the rights of healthcare providers. The contradiction between designated hospitals and non-designated hospitals should be resolved to ensure that PLWHA receive timely and effective help and treatment.

The Toxoplasma gondii ME-49 strain upregulates levels of A20 that inhibit NF-κB activation and promotes apoptosis in human leukaemia T-cell lines.

BACKGROUND: Acute T-lymphocyte leukaemia is a form of haematological malignancy with abnormal activation of NF-κB pathway, which results in high expression of A20 and ABIN1, which constitute a negative feedback mechanism for the regulation of NF-κB activation. Clinical studies have found that acute T-lymphocyte leukaemia patients are susceptible to Toxoplasma gondii infection; however, the effect of T. gondii on the proliferation and apoptosis of human leukaemia T-cells remains unclear. Here, we used the T. gondii ME-49 strain to infect human leukaemia T-cell lines Jurkat and Molt-4, to explore the effect of T. gondii on proliferation and apoptosis, which is mediated by NF-κB in human leukaemia T-cells. METHODS: The Tunel assay was used to detect cell apoptosis. Cell Counting Kit-8 was used to detect cell proliferation viability. The apoptosis level and the expression level of NF-κB related proteins in human leukaemia T-cells were detected by flow cytometry and Western blotting. RESULTS: Western blotting analyses revealed that the T. gondii ME-49 strain increased the expression of A20 and decreased both ABIN1 expression and NF-κB p65 phosphorylation. By constructing a lentiviral-mediated shRNA to knockdown the A20 gene in Jurkat T-cells and Molt-4 T-cells, the apoptosis levels of the two cell lines decreased after T. gondii ME-49 infection, and levels of NF-κB p65 phosphorylation and ABIN1 were higher than in the non-konckdown group. After knockingdown ABIN1 gene expression by constructing the lentiviral-mediated shRNA and transfecting the recombinant expression plasmid containing the ABIN1 gene into two cell lines, apoptosis levels and cleaved caspase-8 expression increased or decreased in response to T. gondii ME-49 infection, respectively. CONCLUSIONS: Our data suggest that ABIN1 protects human leukaemia T-cells by allowing them to resist the apoptosis induced by T. gondii ME-49 and that the T. gondii ME-49 strain induces the apoptosis of human leukaemia T-cells via A20-mediated downregulation of ABIN1 expression.

Trends and Associated Factors of Hypertension among Residents Aged ≥15 Years in Guangzhou, China, 2004-2013.

BACKGROUND: We aimed to describe the trends and associated factors of hypertension among residents aged ≥15 yr in Guangzhou, China. METHODS: Three standardized cross-sectional health surveys were conducted in 2004, 2009 and 2013 using a multi-stage cluster sampling method, and a total of 69128 qualified participants were included in the study. The data were obtained through physical health examination and questionnaire survey. RESULTS: The age-standardised prevalence of hypertension increased from 12.5% to 16.0% between 2004 and 2009 and declined from 16.0% to 14.0% between 2009 and 2013, and crude prevalence respectively was 14.6%, 19.1% and 18.8% in 2004, 2009 and 2013. The proportion of optimal blood pressure dropped from 51.1% to 33.2%, high-normal blood pressure increased from 20.1% to 28.9%, grade 1 hypertension and grade 2 or 3 hypertension increased from 11.5% to 13.6% and 3.9% to 5.8% between 2004 and 2013. The average age was significantly increased (P<0.001) from 42.8 to 47.5 yr, and the average body mass index slightly increased (P<0.001) from 22.4 to 23.0. Logistic regression analysis shows that higher age, male, higher body mass index, smoking and drinking alcohol were potential risk factors for hypertension. CONCLUSION: Both crude and age-standardized prevalence of hypertension were initially increased, but subsequently decreased in Guangzhou during 2004-2013. The optimal blood pressure population decreased significantly while the high-normal blood pressure population increased substantially during the survey period.

Trend and risk factors of low birth weight and macrosomia in south China, 2005-2017: a retrospective observational study.

The percentages of low birth weight (LBW) increased from 7.7% in 2005 to 11.3% in 2011 and declined to 8.1% in 2017. For very low birth weight (VLBW) individuals, the proportion declined -1.0% annually, from 2.5% in 2005 to 1.4% in 2017. Among moderately low birth weight (MLBW) individuals, the proportion first increased 12.8% annually, from 5.0% in 2005 to 9.3% in 2011, and then declined -3.8% annually, from 9.4% in 2011 to 7.0% in 2017. The percentages of macrosomia monotone decreased from 4.0% in 2005 to 2.5% in 2017, an annual decline of -4.0%. Multiple regression analyses showed that boys, maternal age, hypertensive disorders complicating pregnancy (HDCP), and diabetes were significant risk factors for LBW. Boys, maternal age, gestational age, HDCP, diabetes, and maternal BMI were significant risk factors for macrosomia. Although the relevant figures declined slightly in our study, it is likely that LBW and macrosomia will remain a major public health issue over the next few years in China. More research aimed at control and prevention of these risk factors for LBW and macrosomia and their detrimental outcome in the mother and perinatal child should be performed in China.

Time-Dependent Changes in Increased Levels of Plasma Irisin and Muscle PGC-1α and FNDC5 after Exercise in Mice.

Exercise induces the expression of peroxisome proliferator-activated receptor gamma co-activator 1-α (PGC-1α) in skeletal muscle, which promotes the cleavage of fibronectin type III domain-containing protein 5 (FNDC5) to irisin. To explore the relationship between irisin and its regulators, we analyzed the plasma irisin levels and the muscle levels of FNDC5 and PGC-1α after exercise. Male C57BL/6J mice underwent a treadmill exercise (60% of VO2max) for 30 min or one hour (h), and blood and gastrocnemius samples were collected before exercise (pre-exercise), immediately after exercise, and during 24-h recovery after 1-h exercise. We found that plasma irisin levels were significantly increased during exercise (P < 0.05), while FNDC5 protein levels were not significantly increased. Moreover, PGC-1α mRNA and protein levels were significantly increased during 30-min exercise, but were decreased during 1-h exercise. After 1-h exercise, the irisin levels peaked at 6 h (20.71 ± 0.25 ng/ml) and decreased to pre-exercise levels by 24 h (15.45 ± 0.27 ng/ml). Likewise, PGC-1α mRNA and protein levels were increased at 1 h and maintained at elevated levels for 6 h; thereafter, the expression levels of PGC1-α protein were decreased to pre-exercise levels at 12 h. Thus, the restoration of PGC-1α expression to the pre-exercise levels was followed by the decrease in plasma irisin levels. By contrast, during 24-h recovery, the expression levels of FNDC5 mRNA and protein were maintained at elevated levels. These results suggest that the coordinated expression of FNDC5 and PGC-1α may contribute to the increased levels of plasma irisin after exercise.

Medical Discrimination Affects the HIV/AIDS Epidemic Control: A Study of Self-Perceived Medical Discrimination on People Living with HIV or AIDS.

People living with HIV or AIDS (PLWHA) experienced severe medical discrimination which is seriously affecting their lives. However, few studies examined the epidemic characteristics of self-perceived medical discrimination from the discrimination objects such as PLWHA. Therefore, we aimed to investigate the epidemiological status and analyze the influential factors of the self-perceived medical discrimination on PLWHA in South China. The self-designed questionnaire was used to investigate the medical discrimination status of the 443 infected persons, who were randomly recruited from the representative AIDS designated hospitals in Guangdong Province in South China. The results showed that 49.0% of PLWHA experienced medical discrimination, and 55.3% received discriminatory treatment, 48.4% experienced refusal of treatment, 36.4% had private information leaked and 12.9% received mandatory test. However, 52.2% patients chose to endure discrimination in silence. Compared with the Asymptomatic HIV-infected patients, AIDS patients perceived more medical discrimination. The Logistic regression analysis indicated that PLWHA self-perceived medical discrimination status was influenced by 4 factors: the voluntary of first medical detection, the route of transmission, the stage of the disease and the familiarity with the HIV/AIDS-related law. Additionally, the two dimensions of the life quality scale were influenced by medical discrimination, namely, overall function and disclosure worry. Ultimately, our study provides a better understanding of the relationship between infection status, quality of life and the medical discrimination they experienced or perceived. It will help health professionals and policy makers to develop tailored behavioral and policy-oriented intervention strategies for PLWHA to tackle different types of medical discrimination in high-risk settings.