OsmiR396/growth regulating factor modulate rice grain size through direct regulation of embryo-specific miR408.

microRNAs (miRNAs) are promising targets for crop improvement of complex agricultural traits. Coordinated activity between/among different miRNAs may fine-tune specific developmental processes in diverse organisms. Grain size is a main factor determining rice (Oryza sativa L.) crop yield, but the network of miRNAs influencing this trait remains uncharacterized. Here we show that sequestering OsmiR396 through target mimicry (MIM396) can substantially increase grain size in several japonica and indica rice subspecies and in plants with excessive tillers and a high panicle density. Thus, OsmiR396 has a major role related to the regulation of rice grain size. The grain shape of Growth Regulating Factor8 (OsGRF8)-overexpressing transgenic plants was most similar to that of MIM396 plants, suggesting OsGRF8 is a major mediator of OsmiR396 in grain size regulation. A miRNA microarray analysis revealed changes to the expression of many miRNAs, including OsmiR408, in the MIM396 plants. Analyses of gene expression patterns and functions indicated OsmiR408 is an embryo-specific miRNA that positively regulates grain size. Silencing OsmiR408 expression (miR408KO) using CRISPR technology resulted in small grains. Moreover, we revealed the direct regulatory effects of OsGRF8 on OsMIR408 expression. A genetic analysis further showed that the large-grain phenotype of MIM396 plants could be complemented by miR408KO. Also, several hormone signaling pathways might be involved in the OsmiR396/GRF-meditated grain size regulation. Our findings suggest that genetic regulatory networks comprising various miRNAs, such as OsmiR396 and OsmiR408, may be crucial for controlling rice grain size. Furthermore, the OsmiR396/GRF module may be important for breeding new high-yielding rice varieties.

The OsmiR396-OsGRF8-OsF3H-flavonoid pathway mediates resistance to the brown planthopper in rice (Oryza sativa).

Multi-functional microRNAs (miRNAs) are emerging as key modulators of plant-pathogen interactions. Although the involvement of some miRNAs in plant-insect interactions has been revealed, the underlying mechanisms are still elusive. The brown planthopper (BPH) is the most notorious rice (Oryza sativa)-specific insect that causes severe yield losses each year and requires urgent biological control. To reveal the miRNAs involved in rice-BPH interactions, we performed miRNA sequencing and identified BPH-responsive OsmiR396. Sequestering OsmiR396 by overexpressing target mimicry (MIM396) in three genetic backgrounds indicated that OsmiR396 negatively regulated BPH resistance. Overexpression of one BPH-responsive target gene of OsmiR396, growth regulating factor 8 (OsGRF8), showed resistance to BPH. Furthermore, the flavonoid contents increased in both the OsmiR396-sequestered and the OsGRF8 overexpressing plants. By analysing 39 natural rice varieties, the elevated flavonoid contents were found to correlate with enhanced BPH resistance. Artificial applications of flavonoids to wild type (WT) plants also increased resistance to BPH. A BPH-responsive flavanone 3-hydroxylase (OsF3H) gene in the flavonoid biosynthetic pathway was proved to be directly regulated by OsGRF8. A genetic functional analysis of OsF3H revealed its positive role in mediating both the flavonoid contents and BPH resistance. And analysis of the genetic correlation between OsmiR396 and OsF3H showed that down-regulation of OsF3H complemented the BPH resistance characteristic and simultaneously decreased the flavonoid contents of the MIM396 plants. Thus, we revealed a new BPH resistance mechanism mediated by the OsmiR396-OsGRF8-OsF3H-flavonoid pathway. Our study suggests potential applications of miRNAs in BPH resistance breeding.

Modulation of plant architecture by the miR156f-OsSPL7-OsGH3.8 pathway in rice.

Tiller number and plant height are two of the main features of plant architecture that directly influence rice yield. Auxin and miR156, an extensively studied small RNA (smRNA), are both broadly involved in plant development and physiology, suggesting a possible relationship between the two. In this study, we identified a rice T-DNA insertion cluster and dwarf (cd) mutant that has an increased tiller number and reduced plant height. The T-DNA insertion was in close proximity to the miR156f gene and was associated with its up-regulation. Plants overexpressing miR156f resembled the cd mutant. In contrast, plants overexpressing an miR156f target mimic (MIM156fOE) had a reduced tiller number and increased height. Genetic analysis showed that OsSPL7 is a target of miR156f that regulates plant architecture. Plants overexpressing OsSPL7 had a reduced tiller number, while OsSPL7 RNAi plants had an increased tiller number and a reduced height. We also found that OsSPL7 binds directly to the OsGH3.8 promoter to regulate its transcription. Overexpression of OsGH3.8 and OsGH3.8 RNAi partially complemented the MIM156fOE and cd mutant phenotypes, respectively. Our combined data show that the miR156f-OsSPL7-OsGH3.8 pathway regulates tiller number and plant height in rice, and this pathway may allow crosstalk between miR156 and auxin.

[Evaluation of the dietary quality by diet balance index for pregnancy among pregnant women].

OBJECTIVE: To establish the Diet Balance Index for Pregnancy (DBI-P) and utilize it to evaluate the dietary quality among pregnant women in Chengdu, and to provide the basis for maternal nutrition interventions. METHODS: According to the Chinese dietary principles and the nutritional standards (2013) of pregnant women, the Diet Balance Index for Pregnancy (DBI-P) was established on the foundation of the Diet Balance Index (DBI). 610 healthy pregnant women in Chengdu were recruited and were followed up from early pregnancy to the third trimester, and food intakes of them were collected by 24-hour dietary recall method in each trimester. The overall dietary quality in each trimester was evaluated by DBI-P and the variation of dietary quality was analyzed. RESULTS: In each trimester, the intakes of fruits and eggs were optimal, but the meats were over-consumed, and the intakes of cereals, vegetables, milk, soybean and nuts, and fish and shrimp were insufficient. The median total score (TS) was negative in each trimester, suggesting the food intakes of pregnant women were tend to be insufficient. The compliance rate was higher in fruits (59.0% - 79.0%), and lower in the other food groups (5.7% - 48.4%), especially in cereals (5.7% - 7. 4%). In the first, second and third trimester, the median high bound scores (HBS) were 3, 4 and 4, respectively, suggesting the women were in relatively low surplus of food intake, and the degree of over intake were same in the three trimesters (P > 0.05), and the percent of over intake were 21.0%, 33.1% and 24.1%, respectively. The median low bound scores (LBS) were 24, 19 and 21, respectively, suggesting they were in low deficit of food intake, and the degree of under intake was highest in the first trimester (P < 0.017), and the percent of under intake were 96.1%, 88.0% and 91.3%, respectively. The median diet quality distance scores (DQD) were 27, 23 and 25, respectively, suggesting they were in moderate to low dietary imbalance, and the degree of imbalance was highest in the first trimester (P < 0.017), and the percent of overall imbalance were 97.9%, 96.9% and 97.4%, respectively. CONCLUSION: The dietary intake of pregnant women in Chengdu is unbalanced, especially during the first trimester, and the deficit of food intake is the main problem.

[Effect of Plasma Lipids on GDM in the First Trimester of Pregnant Women: a Prospective Cohort Study].

OBJECTIVE: To determine the effect of plasma lipids on gestational diabetes mellitus (GDM) of pregnant women in their first trimester. METHODS: 354 healthy singleton primiparas were followed up in three hospitals in Chengdu until delivery. Basic information about pregnancy was collected using a questionnaire. Dietary intake was assessed using 24 h dietary recall at (12 ± 1) weeks of gestation. Their total energy intake in the first trimester was calculated with a nutrition calculator. Blood samples of the participants were also taken at the (12 ± 1) weeks of gestation to determine plasma triglyceride (TG), total cholesterol (TC), high-density lipoproteincholesterol (HDL-C), low-density lipoproteincholesterol (LDL-C), fasting blood-glucose (FBG). 75 g oral glucose tolerance test (OGTT) was conducted at 24-28 weeks of gestation for the diagnosis of GDM. Multiple logistic regression analyses were performed to estimate odds ratio (OR) of risk factors. Receiver operating characteristic (ROC) curves were drawn to determine the optimal operating point (OOP) of TG level for predicting GDM. Results Women with GDM had significantly higher levels of plasma TG, TC and LDL-C than those without GDM (P < 0.05). After adjustment for maternal pre-pregnant BMI and other confounding factors, women with TG ≥ 1.59 mmol/L and 1.26-1.59 mmol/L showed a higher risk of GDM: 3.86-fold [95% confidence interval (CI): 1.35-11.08] and 2.46-fold (95% CI: 1.05-6.51) as compared with those with TG < 0.94 mmol/L, respectively. The OOP was determined at 1.27 mmol/L with high sensitivity and specificity, and area under the curve 0.634 (95% CI: 0.574-0.711). CONCLUSION: TG level in the first trimester is associated with GDM. It is important to monitor plasma lipids in pregnant women.

[Effect of Sleep in Early Pregnancy on Gestational Diabetes: a Prospective Study.]

OBJECTIVES: To investigate the influence of sleep status in early pregnant women on the occurrence of gestational diabetes mellitus (GDM). METHODS: In this prospective study,542 pregnant women in early pregnancy were selected as subjects.The sleep time,insomnia,pre-gestational body mass and basic information of the subjects were collected by the trained investigators through questionnaire.At the 24th to 28th pregnant week,the body mass of each subject was measured and oral glucose tolerance test (OGTT) was conducted.The diagnosis of GDM was followed the guidelines of diagnose and treatment of gestational diabetes mellitus in China (2014).A non-conditional logistic regression method was adopted to analyze the correlation between sleep status of early pregnant women and the occurrence of GDM. RESULTS: The numbers of subjects in sleep deficiency group,sleep sufficiency group and sleep excessive group were 16 (2.95%),268 (49.45%) and 258 (47.60%),respectively.The incidences of GDM for sleep deficiency group,sleep sufficient group,and sleep excessive group were 62.50%(10/16),25.75%(69/268) and 22.09% (57/258),respectively,the difference was statistically significant (χ2=11.280,P<0.05).Compared with either sleep sufficiency or sleep excessive group,sleep deficiency group had higher GDM occurrence rate with statistically significant (χ2=8.410,11.218,P<0.017).Insomnia subjects in early pregnancy were 137 (25.28%).The incidences of GDM in insomnia group and normal group were 32.12% and 22.72% respectively,the difference was statistically significant (χ2=4.813, P<0.05).After adjusting the confounding factors [age,education,pre-pregnant body mass index (BMI),gained body mass in early and medium pregnancy,family history of type-2 diabetes,gravidity,parity,smoke,drink,exercise and occupation],non-conditional logistic regression results show that sleep deficiency for early pregnant women was an independent risk factor for the occurrence of GDM [odds ratio (OR)=7.38,95% confidence interval (CI): 2.25-24.17].However,the association between insomnia and the occurrence of GDM was not found. CONCLUSIONS: Sleep deficiency in early pregnancy may be a risk factor for the occurrence of GDM.

[Evaluation of pre-pregnancy weight and gestational weight gain among urban and rural women from southwestern China].

OBJECTIVE: To evaluate the situation of pre-pregnancy weight and gestational weight gain (GWG) of women in the urban and rural areas of southwest of China. METHODS: Total 3391 women whose infants and young children aged 6 - 24 months were selected from urban and rural areas of Kunming, Guiyang and Chengdu cities by stratified cluster random sampling. Data of pre-pregnancy height and weight, prenatal weight and pregnancy age for subjects was obtained using a questionnaire. Pre-pregnancy BMI and GWG were calculated. According to the BMI standard for adults from WHO and GWG Guidelines from IOM (2009), the status of pre-pregnancy weight and GWG were assessed. RESULTS: Average BMI of pre-pregnancy for them is (20.3 +/- 2.4). Percentage of normal weight, underweight, and overweight/obesity of pre-pregnancy were 72.7%, 24.1% and 3.2% respectively. The average GWG was (14.9 +/- 6.0) kg, and there was a significant difference between urban and rural group (P < 0.05). Percentage of normal, insufficient and excessive GWG were 35.3%, 31.1% and 33.3% respectively. The rate of excessive GWG in urban group was higher than rural group (P < 0.05), but the rate of insufficient GWG was lower (P < 0.05). The rate of insufficient GWG was higher in women aged below 23 years old (P < 0.05), and the rate of excessive GWG was higher in women aged 24 - 34 years old (P < 0.05). CONCLUSION: It should be pay more attention to improve the underweight of pre-pregnancy and abnormal GWG among women in the southwest of China.

Clinical Outcomes of Severe Rhinosinusitis Complicated with Cavernous Sinus Syndrome.

Background: Various diseases involving the cavernous sinus can cause a condition called cavernous sinus syndrome (CSS), which is characterized by ophthalmoplegia or sensory deficits over the face resulting from the compression effect of internal structure. While tumor compression is the most reported cause of CSS, statistical data on CSS caused by infections are limited. Its risk factors, treatment methods, and clinical outcomes are not well-documented. Methods: In this retrospective study, we reviewed the data of patients admitted to a tertiary medical center from 2015 to 2022 with a diagnosis of acute and chronic sinusitis and at least one diagnostic code for CSS symptoms. We manually reviewed whether patients were involved in two or more of the following cranial nerves (CN): CN III, CN IV, CN V, or CN VI, or at least one of these nerves with a neuroimaging-confirmed lesion in the cavernous sinus. Results: Nine patients were diagnosed with rhinosinusitis-related CSS. The most common comorbidity was type 2 diabetes, and the most common clinical manifestations were diplopia and blurred vision. The sphenoid sinus was the most affected sinus. One patient expired due to a severe brain abscess infection without surgery. The remaining patients underwent functional endoscopic sinus surgery, and 50% of the pathology reports indicated fungal infections. Staphylococcus spp. was the most cultured bacteria, and Amoxycillin/Clavulanate was the most used antibiotic. Only four patients had total recovery during the follow-up one year later. Conclusions: CSS is a rare but serious complication of rhinosinusitis. Patients with diabetes and the elderly may be at a higher risk for this complication. Even after treatment, some patients may still have neurological symptoms.

miR156f integrates panicle architecture through genetic modulation of branch number and pedicel length pathways.

BACKGROUND: Rice (Oryza sativa) panicle architecture is the major determinant of the ideal plant architecture that directly influence yield potential. Many genes influencing development of primary branches, secondary branches, spikelet and pedicel would also influence panicle architecture, which is thus a complex trait regulated by genes from various aspects. miR156, an extensively studied miRNA, has recently emerged as promising target for crop improvement because of its role in plant architecture regulation, such as the number of tillers, plant height and the panicle architecture. Increasing evidence suggests that miR156 might play an important role in panicle architecture regulation. MAIN BODY: To study the detailed function of miR156 in rice panicle architecture regulation, we examined the genetic interaction or transcriptional regulation of miR156/OsSPL to other panicle regulating genes. Our results revealed that expression of many panicle related genes were influenced by miR156. Through biochemical analysis, we further proved that miR156 directly regulated the axillary meristem regulating gene, LAX1, at the transcription level. And the intimate relations between miR156 and LAX1, and miR156 and LAX2 were also uncovered by genetic analysis. On the other hand, a tight genetic linkage between miR156 and RCN2, the panicle branch promoting gene, was also detected, which suggested a buffering mechanism for the miR156 mediated panicle architecture regulation. Furthermore, genetic analysis also demonstrated that miR156 functioned in the same pathway with OsRA2 to regulate pedicel length. SHORT CONCLUSION: Altogether, miR156 integrates several genetic pathways mediated by genes such as LAX1, LAX2, RCN2 and OsRA2, and comprehensively regulates panicle development in rice. Based on these analysis, we concluded that miR156 acts as an important regulator for panicle architecture through influencing various aspects of panicle development.

The doctor's digital double: how warmth, competence, and animation promote adherence intention.

BACKGROUND: Each year, patient nonadherence to treatment advice costs the US healthcare system more than $300 billion and results in 250,000 deaths. Developing virtual consultations to promote adherence could improve public health while cutting healthcare costs and usage. However, inconsistencies in the realism of computer-animated humans may cause them to appear eerie, a phenomenon termed the uncanny valley. Eeriness could reduce a virtual doctor's credibility and patients' adherence. METHODS: In a 2 × 2 × 2 between-groups posttest-only experiment, 738 participants played the role of a patient in a hypothetical virtual consultation with a doctor. The consultation varied in the doctor's Character (good or poor bedside manner), Outcome (received a fellowship or sued for malpractice), and Depiction (a recorded video of a real human actor or of his 3D computer-animated double). Character, Outcome, and Depiction were designed to manipulate the doctor's level of warmth, competence, and realism, respectively. RESULTS: Warmth and competence increased adherence intention and consultation enjoyment, but realism did not. On the contrary, the computer-animated doctor increased adherence intention and consultation enjoyment significantly more than the doctor portrayed by a human actor. We propose that enjoyment of the animated consultation caused the doctor to appear warmer and more real, compensating for his realism inconsistency. Expressed as a path model, this explanation fit the data. DISCUSSION: The acceptance and effectiveness of the animation should encourage the development of virtual consultations, which have advantages over creating content with human actors including ease of scenario revision, internationalization, localization, personalization, and web distribution.

OsRAMOSA2 Shapes Panicle Architecture through Regulating Pedicel Length.

The panicle architecture of rice is an important characteristic that influences reproductive success and yield. It is largely determined by the number and length of the primary and secondary branches. The number of panicle branches is defined by the inflorescence meristem state between determinacy and indeterminacy; for example, the maize ramosa2 (ra2) mutant has more branches in its tassel through loss of spikelet determinacy. Some genes and factors influencing the number of primary and secondary branches have been studied, but little is known about the molecular mechanism underlying pedicel development, which also influences panicle architecture. We report here that rice OsRAMOSA2 (OsRA2) gene modifies panicle architecture through regulating pedicel length. Ectopic expression of OsRA2 resulted in a shortened pedicel while inhibition of OsRA2 through RNA interference produced elongated pedicel. In addition, OsRA2 influenced seed morphology. The OsRA2 protein localized to the nucleus and showed transcriptional activation in yeast; in accordance with its function in pedicel development, OsRA2 mRNA was enriched in the anlagen of axillary meristems, such as primary and secondary branch meristems and the spikelet meristems of young panicles. This indicates a conserved role of OsRA2 for shaping the initial steps of inflorescence architecture. Genetic analysis revealed that OsRA2 may control panicle architecture using the same pathway as that of the axillary meristem gene LAX1 (LAX PANICLE1). Moreover, OsRA2 acted downstream of RCN2 in regulating pedicel and branch lengths, but upstream of RCN2 for control of the number of secondary branches, indicating that branch number and length development in the panicle were respectively regulated using parallel pathway. Functional conservation between OsRA2 and AtLOB, and the conservation and diversification of RA2 in maize and rice are also discussed.

Sleep disturbances during pregnancy are associated with cesarean delivery and preterm birth.

OBJECTIVE: The purpose of this study was to examine the associations of sleep disturbances during pregnancy with cesarean delivery and preterm birth. METHODS: In this prospective study, 688 healthy women with singleton pregnancy were selected from three hospitals in Chengdu, China 2013-2014. Self-report questionnaires, including the sleep quantity and quality as well as exercise habits in a recent month were administered at 12-16, 24-28, and 32-36 weeks' gestation. Data on type of delivery, gestational age, and the neonates' weight were recorded after delivery. After controlling the potential confounders, a serial of multi-factor logistic regression models were performed to evaluate whether sleep quality and quantity were associated with cesarean delivery and preterm birth. RESULTS: There were 382 (55.5%) women who had cesarean deliveries and 32 (4.7%) who delivered preterm. Women with poor sleep quality during the first (OR: 1.87, 95% CI [1.02-3.43]), second (5.19 [2.25-11.97]), and third trimester (1.82 [1.18-2.80]) were at high risk of cesarean delivery. Women with poor sleep quality during the second (5.35 [2.10-13.63]) and third trimester (3.01 [1.26-7.19]) as well as short sleep time (<7 h) during the third trimester (4.67 [1.24-17.50]) were at high risk of preterm birth. CONCLUSIONS: Sleep disturbances are associated with an increased risk of cesarean delivery and preterm birth throughout pregnancy. Obstetric care providers should advise women with childbearing age to practice healthy sleep hygiene measures.

OsMADS1 Represses microRNA172 in Elongation of Palea/Lemma Development in Rice.

Specification of floral organ identity is critical for the establishment of floral morphology and inflorescence architecture. Although multiple genes are involved in the regulation of floral organogenesis, our understanding of the underlying regulating network is still fragmentary. MADs-box genes are principle members in the ABCDE model that characterized floral organs. OsMADS1 specifies the determinacy of spikelet meristem and lemma/palea identity in rice. However, the pathway through which OsMADS1 regulates floral organs remains elusive; here, we identified the microRNA172 (miR172) family as possible regulators downstream of OsMADS1. Genetic study revealed that overexpression of each miR172 gene resulted in elongated lemma/palea and indeterminacy of the floret, which resemble the phenotype of osmads1 mutant. On the contrary, overexpression of each target APETALA2 (AP2) genes resulted in shortened palea/lemma. Expression level and specificity of miR172 was greatly influenced by OsMADS1, as revealed by Northern blot analysis and In situ hybridization. Genetically, AP2-3 and AP2-2 over expression rescued the elongation and inconsistent development of the lemma/palea in OsMADS1RNAi transgenic plants. Our results suggested that in rice, OsMADS1 and miR172s/AP2s formed a regulatory network involved in floral organ development, particularly the elongation of the lemma and the palea.

[The effect of pre-pregnancy body mass index, gestational weight gain on the onset of lactation: a longitudinal study].

OBJECTIVE: To explore the effect of pre-pregnancy body mass index (BMI) and gestational weight gain (GWG) on the onset time of lactation (OL). METHODS: In this longitudinal study, 751 healthy single pregnant women were selected from three hospitals in Chengdu as subjects of baseline survey during March and September 2013 and were followed before and after delivery, respectively. Finally, data from a total of 473 pregnant women were analyzed. Data on pre-pregnancy weight, delivery mode, birth weight of neonate, information of lactation and maternal demographic characteristics were collected through questionnaire and medical records. Height and weight at the last week before delivery were measured and GWG were calculated. After controlling the potential confounders, a serial of multi-factor ordinal logistic regression models were performed to test the associations between pre-pregnancy BMI, GWG and OL. RESULTS: The proportions of OL at the < 1, 1-, 24-, 48- and ≥ 72 h groups were 16.3%, 37.0%, 17.5%, 18.6% and 10.6%, respectively. When compared to women with normal weight, the ORs for prolonged OL were 2.85 (1.91-4.27) and 3.42 (1.69-6.90) among pre-pregnant underweight and overweight/obese women, respectively. When compared to women with normal weight and adequate GWG, pre-pregnant underweight women with adequate and excessive GWG showed greater odds of prolonged OL (OR = 2.34, 95% CI: 1.31-4.18; OR = 3.42, 95% CI: 1.67-7.00), respectively. Pre-pregnant overweight/obese women with excessive GWG had increased the odds of prolonged OL (OR = 3.10, 95% CI: 1.15-8.37). CONCLUSION: Pre-pregnant BMI appeared an independently factor, associated with OL while GWG might have an effect on OL, when pre-pregnant BMI was considered.