Rhamnocitrin decreases fibrosis of ovarian granulosa cells by regulating the activation of the PPARγ/NF-κB/TGF-ß1/Smad2/3 signaling pathway mediated by Wisp2.

Background: Polycystic ovary syndrome (PCOS) is the most common cause of anovulatory infertility in women. Rhamnocitrin (Rha) has anti-inflammatory and antioxidant actions. The WNT1-inducible-signaling pathway protein 2 (Wisp2) and nuclear factor (NF)-κB are involved in fibrosis in many diseases. We aimed to elucidate the role of Rha in fibrosis of PCOS and the underlying mechanisms. Methods: Dehydroepiandrosterone (DHEA)-incubated ovarian granulosa KGN cells were treated by Rha. Cell proliferation was detected with cell counting kit-8 (CCK-8) and 5-ethynyul-2'-deoxyuridine (EdU) staining. The levels of Wisp2 and transforming growth factor-ß1 (TGF-ß1) in supernatant were measured by enzyme-linked immunosorbent assay (ELISA). We observed α-smooth muscle actin (α-SMA) protein by immunofluorescence (IF). The levels of fibrosis factors were determined using Western blot. We observed p65 nuclear translocation with confocal microscopy. We used Wisp2 overexpression and knockdown in cells treated with DHEA or Rha to validate Wisp2 function. Interaction between Wisp2 and NF-κB, as well as Wisp2 and PPARγ, were assessed by co-immunoprecipitation assay, luciferase reporter assay and chromatin immunoprecipitation (ChIP). Results: The results showed that Rha elevated the reduced proliferation of DHEA-treated cells. In addition, Rha reversed the decreased Wisp2 and the increased TGF-ß1 in supernatant. The proteins CTGF, α-SMA, Collagen I, TGF-ß1, p-Smad2, and p-Smad3 were up-regulated while Wisp2, Sirt1, and PPARγ were down-regulated by DHEA treatment, which were reversed by Rha. Meanwhile, DHEA up-regulated p-IKBa and p-p65 and promoted p65 nuclear translocation, which were inhibited by Rha. These effects of Rha were antagonized by Wisp2 knockdown and were mimicked by Wisp2 overexpression. We confirmed the protein interaction between Wisp2 and NF-κB, along with Wisp2 and PPARγ. Conclusions: Wisp2-mediated PPARγ/NF-κB/TGF-ß1/Smad2/3 signaling contributes to Rha-improved ovarian granulosa cells fibrosis, suggesting Rha as a novel agent for the treatment of PCOS.

[Under-5-mortality rate and causes of death in China, 2000 to 2010].

OBJECTIVE: To assess the changes and the leading cause of deaths for children under 5 years old, in China, during 2000 - 2010, with the aim of evaluation on the progress in achieving the relative goal set by "National Program of Action for Child Development in China (2001 - 2010)", and understanding the related challenges. METHODS: Data used in this study were collected from the population-based National Maternal and Child's Health Surveillance Network of China. Infant Mortality Rate (IMR), Under-5-mortality rate (U5MR) and the leading cause of deaths for under-5 children were analyzed. RESULTS: Nationwide IMR and U5MR in 2010 dropped by 59.3% and 58.7% respectively, compared to that in 2000. Decreases by 50.8% and 47.1% in IMR and U5MR were observed in urban areas, and 56.5% and 56.0% in rural areas during this period. Compared with data from 2000, the leading causes-specific U5MR in 2010 had significantly declined. The top 5 leading causes of death in 2010 were premature birth/low birth weight, pneumonia, birth asphyxia, congenital heart disease and accidental suffocation, but were different in urban and rural areas. In 2010, both IMR and U5MR from the rural areas were 2.8-folds than that of the urban areas. In addition, IMRs in the Middle and Western parts of China were 1.5 and 2.3-folds respectively of that in the East, and U5MR in Middle and West was 1.5 and 2.2-folds respectively of that in East. CONCLUSION: IMR, U5MR and the leading causes specific mortality rate in China declined remarkably from 2000 to 2010, and the goal set by "National Program of Action for Child Development in China (2001 - 2010)" had been successfully achieved. However, the disparity on child's health in regions and in urban or rural areas, still remained a challenge.

[Analysis on the diarrhea mortality rate, pre-death diagnosis and treatment of children under-5 in China, 1996-2006].

OBJECTIVE: To understand the trends of diarrhea mortality rate, pre-death diagnosis and treatment of children under-5 in China, from 1996 to 2006. METHODS: We used data obtained from the 1996 to 2006 national child mortality surveillance network, including 116 counties (cities) throughout China, to evaluate the under 5 mortality rate (U5MR) due to diarrhea in different geographical areas, and related factors of under 5 children mortality due to diarrhea. RESULTS: Data from the national U5MR due to diarrhea fell from 249.8 in 1996 to 75.6 per 100 000 live births, in 2006. The U5MR due to diarrhea in urban reduces from 11.6 in 1996 to 6.1 per 100 000 live births in 2006, with a reduction of 47.4%. The U5MR due to diarrhea in rural decreased from 304.7 in 1996 to 94.3 per 100 000 live births in 2006, with a reduction of 69.1%. The U5MR due to diarrhea in coastal, inland and remote areas fell from 48.9, 178.9 and 566.9 in 1996 to 6.2, 30.4 and 199.2 per 100 000 live births in 2006, with a reduction of 87.3%, 83.0% and 64.9%, respectively. Among the rural children died of diarrhea, about 37.9% were diagnosed in a village clinic, 15.1% never received any diagnostic procedure. Nearly 20% of the patients had not been treated, with 50% - 60% of them had only been treated in an outpatient department. CONCLUSION: During 1996 - 2006, the U5MR due to diarrhea showed a substantially downward trend in China but the disparities between urban and rural, remote and coastal, areas were increasing.

[Mortality from congenital malformations in children under 5 years old in China, 1996-2006].

OBJECTIVE: To identify the trend and pattern of mortality from congenital malformations in children under 5 years old in China. METHODS: Data from 1996 to 2006 were obtained from the national child mortality surveillance network, which included 116 counties (cities) throughout China. A descriptive analysis was performed on birth defect-specific mortality (U5MR) in children under 5 years old. RESULTS: The national birth defect-specific U5MR decreased from 412.0 per 100,000 live births in 1996 to 342.2 per 100,000 live births in 2006. The declining trend in birth defect-specific U5MR was more obvious from 2003 to 2006. The same pattern was observed in urban areas, in coastal regions and in inner land regions of China. Rural areas had higher birth defect-specific U5MR than urban areas. Remote regions had higher birth defect-specific U5MR than coastal regions and inner land regions. The proportion of mortality due to congenital malformations in total U5MR increased from 9.2% in 1996 to 14.4% in 2006. CONCLUSION: The proportion of mortality due to congenital malformations in children under 5 years old is increasing in China.

[Analysis on under-5 mortality rate and the leading kinds of diseases in China, from 2000 to 2006].

OBJECTIVE: To study the changing pattern of infant mortality and under-5 mortality rate in China from 2000 to 2006, and to evaluate China's progress in achieving the United Nations' Millennium Development Goal 4. METHODS: A population-based survey was conducted through a nationwide multi-level surveillance network. The mortality rate and the proportion of death for children under 5 were analyzed. RESULTS: The infant mortality rate (IMR), under-5 mortality rate (U5MR) in China dropped to 17.2, 20.6 per 1000 live births in 2006, respectively, comparing to 32.2 and 39.7 per 1000 live births in 2000. In urban areas, IMR, U5MR dropped to 8.0, 9.6 per 1000 live births in 2006, respectively while they were 11.8 and 13.8 per 1000 live births respectively in 2000. In rural areas, IMR, U5MR dropped to 19.7 and 23.6 per 1000 live births in 2006, respectively but they were 37.0 and 45.7 per 1000 live births respectively in 2000. During this period, the mortality rates due to pneumonia and diarrhea had dropped sharply. The proportion of deaths due to pneumonia, diarrhea also dropped from 19.5%, 4.9% in 2000 to 15.6%, 3.7% in 2006, respectively. In urban areas, the proportion of deaths due to pneumonia dropped from 9.9% in 2000 to 9.8% in 2006, In rural areas, the proportion of deaths due to pneumonia, diarrhea dropped from 20.1%, 5.2% in 2000 to 16.2%, 4.0% in 2006, respectively. CONCLUSION: The U5MR in China remarkably dropped from 2000 to 2006. Based on data through the surveillance program, China should be able to accomplish the Millennium Development Goals 4 of the United Nations as planned.

An essential role for the RNA-binding protein Smaug during the Drosophila maternal-to-zygotic transition.

Genetic control of embryogenesis switches from the maternal to the zygotic genome during the maternal-to-zygotic transition (MZT), when maternal mRNAs are destroyed, high-level zygotic transcription is initiated, the replication checkpoint is activated and the cell cycle slows. The midblastula transition (MBT) is the first morphological event that requires zygotic gene expression. The Drosophila MBT is marked by blastoderm cellularization and follows 13 cleavage-stage divisions. The RNA-binding protein Smaug is required for cleavage-independent maternal transcript destruction during the Drosophila MZT. Here, we show that smaug mutants also disrupt syncytial blastoderm stage cell-cycle delays, DNA replication checkpoint activation, cellularization, and high-level zygotic expression of protein coding and micro RNA genes. We also show that Smaug protein levels increase through the cleavage divisions and peak when the checkpoint is activated and zygotic transcription initiates, and that transgenic expression of Smaug in an anterior-to-posterior gradient produces a concomitant gradient in the timing of maternal transcript destruction, cleavage cell cycle delays, zygotic gene transcription, cellularization and gastrulation. Smaug accumulation thus coordinates progression through the MZT.

[DNA mismatch repair enzyme hMSH2 genetic polymorphism in southern Chinese Han population].

OBJECTIVE: To study hMSH2 genetic polymorphism in southern Chinese Han population. METHODS: The basic materials and blood samples from 163 southern Chinese were collected. The mutations of exon 6 and exon 7 of hMSH2 gene were investigated by PCR-SSCP, followed by DNA sequencing. RESULTS: Fragments of 250 bp including exon 6 and fragments of 323 bp including exon 7 of hMSH2 gene were amplified by multiple PCR. The allele frequencies of C18, A82 and B39 type mutations were 0.0184, 0.0031, 0.0031, respectively. The gene frequencies and gene type frequencies of three polymorphism sites in normal population accorded with Hardy-Weinberg equilibrium (P>0.05). The heterozygosity of C18 type mutation (0.0361) was the highest. CONCLUSION: There were three polymorphism sites in exon 7 of hMSH2 gene in southern Chinese Han population, among which the genotype frequency of C18 type was the highest, suggesting that C18 type mutation be a useful genetic mark.