The H1/H5 domain contributes to OsTRBF2 phase separation and gene repression during rice development.

Transcription factors (TFs) tightly control plant development by regulating gene expression. The phase separation of TFs plays a vital role in gene regulation. Many plant TFs have the potential to form phase-separated protein condensates; however, little is known about which TFs are regulated by phase separation and how it affects their roles in plant development. Here, we report that the rice (Oryza sativa) single Myb TF TELOMERE REPEAT-BINDING FACTOR 2 (TRBF2) is highly expressed in fast-growing tissues at the seedling stage. TRBF2 is a transcriptional repressor that binds to the transcriptional start site of thousands of genes. Mutation of TRBF2 leads to pleiotropic developmental defects and misexpression of many genes. TRBF2 displays characteristics consistent with phase separation in vivo and forms phase-separated condensates in vitro. The H1/H5 domain of TRBF2 plays a crucial role in phase separation, chromatin targeting and gene repression. Replacing the H1/H5 domain by a phase-separated intrinsically disordered region from Arabidopsis (Arabidopsis thaliana) AtSERRATE partially recovers the function of TRBF2 in gene repression in vitro and in transgenic plants. We also found that TRBF2 is required for trimethylation of histone H3 Lys27 (H3K27me3) deposition at specific genes and genome-wide. Our findings reveal that phase separation of TRBF2 facilitates gene repression in rice development.

Association between ageing knowledge and willingness to care for older adults among nursing students in China: the mediating role of attitude towards older adults.

Little is known about the mediating effect of attitude toward older adults on the relationship between aging knowledge and willingness to care for older adults. We applied the theory of planned behavior (TPB) and the knowledge-attitude-behavior (KAB) model as theoretical frameworks to examine the mediation effect of attitude toward older adults. Data from 388 Chinese nursing students were analyzed. The Willingness to Care for Older People (WCOP) scale, Kogan's Attitude toward Older People scale (KAOP) and Facts on Aging Quiz (FAQ) were utilized to assess willingness, attitude and aging knowledge, respectively. Data were analyzed using SPSS 22. 0 with the PROCESS macro. Bootstrap methods were used to obtain the significance of mediating effects. The study showed that aging knowledge was significantly associated with willingness to care for older adults and that attitude toward older adults mediated the association. Bootstrapping method confirmed the significance of the indirect effect of aging knowledge through attitude, accounting for 18.9% of the total willingness variance. Overall, based on the TPB and the KAB theoretical framework, our data support the notion that improving aging knowledge and attitude may contribute to improve the willingness to aged care among nursing students.

Verification of data in a nationwide transanal total mesorectal excision registry in China.

BACKGROUND AND OBJECTIVES: Transanal total mesorectal excision is a surgical procedure for mid- and low rectal cancer. The Chinese TaTME Registry Collaborative is a nationwide database collecting information on patients who have undergone this procedure. METHODS: Centers were invited by the registry committee to participate in a three-part data audit project: remote audits for data completeness and deviation values, onsite source verification of data accuracy, and an online survey of the characteristics of data managers. RESULTS: Twenty-three tertiary centers participated in this project. The median case volume registered by the centers was 51 (interquartile range, 25-89). The overall data completeness for 30 verified variables was 89.1%. Eight centers achieved a high data completeness rate (>95%). The source data of eight centers were verified onsite. The overall accuracy rate was 90.4% (85.3%-97.6% across centers). Postoperative complications, mortality, and proximal/distal resection margin involvement were accurately reported in >95% of cases. The data completeness rate was higher if the data manager was a surgeon/surgical resident (94.2% vs. 84.8%, p = 0.045). CONCLUSIONS: The completeness and accuracy of the data in the Chinese TaTME Registry Collaborative are acceptable. The quality of the data is highest when entered by colorectal surgeons and residents.

Phospholipidase Dδ Negatively Regulates the Function of Resistance to Pseudomonas syringae pv. Maculicola 1 (RPM1).

RPM1 is a plant immune receptor that specially recognizes pathogen-released effectors to activate effector-triggered immunity (ETI) in Arabidopsis thaliana. RPM1 triggers ETI and hypersensitive response (HR) for disease resistance. Previous reports indicated that Phospholipase D (PLD) positively regulated RPM1-mediated HR. However, single, double, and triple pld knock-out mutants of 12 members of the PLD family in A. thaliana did not show suppressed RPM1-mediated HR, indicating the functional redundancy among PLD members. In this study, we revealed that PLD could negatively regulate the function of RPM1. We found that RPM1 interacted with PLDδ, but did not interact with PLDß1, PLDß2, and PLDγ3. Overexpression of PLDδ conducted to a reduction of protein level and corresponding activity of RPM1. We found that abscisic acid (ABA) reduced the protein level of RPM1, and the ABA-induced RPM1 reduction required PLD activity and PLD-derived phosphatidic acid (PA). Our study shows that PLD plays both negative and positive roles regulating the protein level and activity of RPM1 during stress responses in plants. PLD proteins are regulating points to integrate the abiotic and biotic responses of plants.

Functional analysis of plant NB-LRR gene L3 by using E. coli.

Plant NB-LRR genes mediate plant innate immunity and cause the programmed cell death of plant cells. Very little, however, is known about these processes. Taken advantage of easy manipulation of bacteria, genetic analysis was made to understand the mechanism of lethality of NB-LRR proteins to bacteria and correlate the information back to how NB-LRR proteins cause cell death in plants. It was found that only L3 encoded by NB-LRR gene L3 (At1g15890) specifically caused significant death of BL21(DE3), while other NBS-LRR proteins did not, and 760-851, the truncated form of L3, was essential to the lethality of L3. Gene yedZ (EG14048) and nupG (EG10664) were identified by genome re-sequencing from E. coli, both of which mediate the toxicity of L3 in E. coli. Furthermore, NupG can affect the activity of peroxidase and significantly suppress plant cell death, which is induced by NB-LRR protein RPM1(D505V) encoded by RPM1 (At3g07040) in N. benthamiana. These findings provide evidence that functional analysis of plant NB-LRR genes in microorganisms might be a potential and rapid method.

Risk factors for measles in children younger than age 8 months: A case-control study during an outbreak in Guangxi, China, 2013.

BACKGROUND: Following a period (2009-2012) during which zero measles cases were reported, a measles outbreak occurred in 2013 in Bama County, Guangxi, China, that involved more than 100 children younger than age 8 months. We aimed to identify the pitfalls and risk factors while implementing the control measures. METHODS: An outbreak investigation and a case-control study was conducted among children younger than age 8 months. The serum specimens of the study subjects and their mothers were tested for measles immunoglobulin M and immunoglobulin G. RESULTS: The attack rate was 2.3/1,000 population. The median (interquartile range) age was 18.6 months (7.9-52.8 months). The coverage of 2-dose measles-containing vaccine was only 34%. The case-control study revealed 2 independent risk factors: low education level of main caregiver (odds ratio [OR], 2.86; 95% confidence interval [CI], 1.31-6.22) and visiting a hospital 7-21 days before the date of symptoms onset (OR, 9.84; 95% CI, 4.27-22.67). The population attributable fraction of the latter was 52.8%. The mothers of the cases had nonsignificantly higher levels of immunoglobulin M and were significantly more likely to have protective levels of immunoglobulin G than those of the controls. This suggests a reactive rather than protective role of the antibody to the child's infection. CONCLUSIONS: In a near-elimination but low measles-containing vaccine coverage community, supplementary immunization activities should be emphasized for children and women who are potential future mothers. The minimum age of measles-containing vaccine should be further reduced. Hospital measles transmission must also be strictly prevented.

Functionality of disintegrants and their mixtures in enabling fast disintegration of tablets by a quality by design approach.

Investigation of the effect of disintegrants on the disintegration time and hardness of rapidly disintegrating tablets (RDTs) was carried out using a quality by design (QbD) paradigm. Ascorbic acid, aspirin, and ibuprofen, which have different water solubilities, were chosen as the drug models. Disintegration time and hardness of RDTs were determined and modeled by executing combined optimal design. The generated models were validated and used for further analysis. Sodium starch glycolate, croscarmellose sodium, and crospovidone were found to lengthen disintegration time when utilized at high concentrations. Sodium starch glycolate and crospovidone worked synergistically in aspirin RDTs to decrease disintegration time. Sodium starch glycolate-crospovidone mixtures, as well as croscarmellose sodium-crospovidone mixtures, also decreased disintegration time in ibuprofen RDTs at high compression pressures as compared to the disintegrants used alone. The use of sodium starch glycolate in RDTs with highly water soluble active ingredients like ascorbic acid slowed disintegration, while microcrystalline cellulose and crospovidone drew water into the tablet rapidly and quickened disintegration. Graphical optimization analysis demonstrated that the RDTs with desired disintegration times and hardness can be formulated with a larger area of design space by combining disintegrants at difference compression pressures. QbD was an efficient and effective paradigm in understanding formulation and process parameters and building quality in to RDT formulated systems.

Clinical application of ultrasound three perpendicular planes plus special planes in diagnosis of fetal cleft lip/palate / 中华医学超声杂志（电子版）

Objective To study the clinical significance of the method of three perpendicular planes plus special planes in diagnosing fetal cleft lip /palate by prenatal ultrasound .Methods The approach of three perpendicular planes and special planes were used in diagnosing 110 cases of cleft lip/palate.The sonogram features in each section were analyzed and the outcomes were recorded during follow-up.Results On prenatal ultrsound ,110 cases were examined with three perpendicular planes method .The coronary section could be displayed at 100%cases (110 cases), sagittal section 76.4%cases (84 cases),transverse section 96.4%cases (106 cases) and parasagittal section 25.5%cases (28 cases).With special planes method,74 cases were examined .The section through pyriform aperture could be displayed in 47 cases,in 45 cases on the section through the lower lip/lower jaw/submandibular triangle ,and in 16 cases on the section through the cheek.Combining the three perpendicular planes and special planes methods ,94.5%(104/110) cases could be diagnosed definitely.Six cases (5.5%,6/110) were missed because of fetal position or oligoamnios . Conclusions The method of three perpendicular planes plus special planes is effective in prenatal ultrasound diagnosing cleft lip/palate,which is of great help in improving prenatal diagnostic accuracy of fetal cleft lip/palate.

Echocardiographic abnormalities in a cohort of Chinese patients with systemic lupus erythematosus--a retrospective analysis of eighty-five cases.

OBJECTIVE: To investigate echocardiographic abnormalities in a cohort of Chinese patients with systemic lupus erythematosus (SLE). METHODS: Eighty-five SLE patients who were hospitalized at our institution from January 2008 to June 2009 were included. Their clinical manifestations, electrocardiogram, chest x-ray, CT scan, and echocardiographic data were analyzed in relation to age, disease duration, and SLE disease activity index (SLEDAI). RESULTS: Echocardiography showed abnormalities in 53 patients (62.35%), involving valves in 56.5% (n = 48), pericardium in 25.9%, left and right ventricle enlargement in 17.65% and 1.18%, respectively, and auricle in 10.59% and 1.18% of the patients, respectively. The left ventricle showed decreased compliance in 10.29%, impaired relaxation in 14.12%, and abnormal wall motion in 42.35% of the patients. Interventricular septum thickness, left ventricular posterior wall thickness, and isovolumic relaxation time were positively correlated with age (r = 0.42, 0.33, and 0.39, respectively, p < 0.01 for all variables), and disease duration (r = 0.22, 0.21, and 0.32, respectively; p < 0.05). Left atrial and ventricular size were correlated with age (r = 0.32 and 0.41, p > 0.01) and SLEDAI (p < 0.05). CONCLUSIONS: Cardiac, especially valvular abnormalities are common in SLE patients and increase with age and disease duration and activity. Inclusion of echocardiography in the clinical evaluation of SLE patients could help identify an important subset of patients with cardiac abnormalities.

Isolation and genetic analysis of a novel triple-reassortant H1N1 influenza virus from a pig in China.

Influenza A viruses of subtype H1N1 have been reported widely in pigs in China, associated with clinical disease. These mainly include classical swine H1N1, avian-like H1N1, and human-like H1N1 viruses. In this study, we reported a novel triple-reassortant H1N1 virus (A/swine/Guangdong/1/2010) containing genes from the classical swine (NP, NS), human (PB1) and avian (HA, NA, M, PB2, PA) lineages, which was for the first time reported in China. Also, phylogenetic analysis further confirmed that five genes segments (NS, NP, PB2, PB1, PA) of the isolate were closely related to the novel reassortant H1N2 viruses isolated in China in 2006, while the other three (HA, NA, M) were closely related to avian-like H1N1 viruses in China. The isolation of triple-reassortant H1N1 influenza virus provides further evidence that pigs serve as emergence hosts or "mixing vessels", and swine influenza virus (SIV) surveillance in China should be given a high priority.

Seroprevalence of rabies virus antibodies in bats from southern China.

Members of the Order Chiroptera are the natural reservoirs of lyssaviruses and play an important role in the transmission of rabies to animals and humans. In this present study, the seroprevalence for rabies virus was determined for bats sampled from four southern provinces on the Chinese mainland. A total of 685 bats of 8 species representing 4 families were collected from 10 sites, and were tested by the indirect fluorescent antibody test using fluorescein isothiocyanate (FITC)-conjugated protein A/G mixture and viral neutralization test. Rabies antibody response was only detected from three bat species (Rousettus leschenaulti, Rhinolophus blythi, and Rhinolophus ferrumequinum). The overall rabies seroconversion rate was only 2.2% (15/685). Of the 15 positive sera, 13 (12 fruit bats and 1 insectivorous bat) were indirect fluorescent antibody test positive, and two insectivorous bats were virus neutralization positive when tested by the modified fluorescent antibody viral neutralization test, albeit extremely low. To our knowledge, this is the first published report describing rabies seroprevalences from Chinese bats. These results suggest that bats may play a role in the ecology of lyssaviruses in China, and further surveillance for the presence of lyssaviruses in bats should be undertaken throughout the country and extended to other species.

Effect of hepatocyte growth factor gene transfection on biological features of lymphoma cells / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the biological effect of hepatocyte growth factor (HGF) on HGF gene-transfected Raji cells.</p><p><b>METHODS</b>Total RNA was extracted from human hepatic tissue, HGF gene cDNA was amplified by RT-PCR, and then cloned into vector pVITRO2-mcs to construct recombinant eukaryotic expression vector pVITRO2-mcs-HGF. The recombinant vector was transfected to Raji cells, and the stably transfected cells were selected by homomycin B in serial passages, and confirmed by real-time fluorescent quantitative PCR, ELISA, immunocytohistochemistry. The biological features of transfected Raji cells were evaluated by semisolid culture.</p><p><b>RESULTS</b>RT-PCR results showed that Raji cells were transfected successfully with recombinant eukaryotic expression vector pVITRO2-mcs-HGF. HGF mRNA and protein were expressed successfully in Raji cells. Expression of HGF gene enhanced proliferation, metastasis and invasion of Raji cells.</p><p><b>CONCLUSION</b>HGF gene has been cloned and recombined to construct recombinant eukaryotic expression vector pVITRO2-mcs-HGF successfully. Transfected HGF may change the biological features of Raji cells.</p>

[Morphological comparison of rabies viral particles before and after rejuvenation in suckling mice and morphogenesis of the rejuvenated virus in cells].

OBJECTIVE: To observe the morphological recovery of rabies virus strain SRV 9 after rejuvenation in suckling mice and to study its morphogenesis in BHK-21 cells. METHODS: The long freeze-preserved rabies virus strain SRV 9 was rejuvenated through intracerebral inoculation of sucking mice twice, followed by propagation in BHK-21 cells. After cell culture the virus was purified through sucrose gradient density ultracentrifugation. RESULTS: Electromicroscopy of the purified virus showed that effective recovery of viral shape was obtained after the rejuvenation with majority of viral particles having a typical bullet-like shape and intact spikes on viral membrane. The proportion of DI particles (with short triangle and irregular shapes) in rejuvenatd virus supernatant was significantly decreased compared to un-rejuvenated virus. Viral morphogenesis in cells showed that typical virus particles could form in intracytoplasm 24 hours p.i. and the number of matured viral particles in cytoplasm increased significantly as culture was prolonged from 24 hours to 96 hours p.i.. Furthermore, the rejuvenated virus was observed budding from vacuole membrane in different patterns. CONCLUSION: (1) The proportion of DI particles can be significantly decreased by rejuvenated through intracerebral inoculation of sucking mice.(2) The optimal harvest opportunity of SRV9 is after being 1-2 undiluted passaged. (3) This research provided more information about morphogenesis of rabies virus.

[Expression of Nipah virus structural proteins F1 and G and preparation of hyperimmune antisera against two proteins].

The fusion protein (F) and attachment glycoprotein (G) of Nipah virus (NiV) are important for the virus to infect cells and induce protective immunity. In this study, the NiV F1 and G gene fragments without the sequences of signal peptide and transmembrane domain were amplified by PCR, then cloned into E. coli expression vector pGEX-6P-1 and modified baculovirus vector, respectively. After induction by IPTG, NiV F1 and G proteins were efficiently expressed in E. coli when analyzed by SDS-PAGE, both showing good reactivity with the rabbit antiserum anti-NiV serum in Western blot. The expression of NiV F1 and G in baculovirus system were also detected by indirect immunofluorescent assay (IFA) of fixed Sf9 cells monolayer infected with the recombinant baculoviruses expressing F1 and G. Furthermore the anti-F1 and anti-G hyperimmune sera were prepared by immunization of rabbits respectively with purified E. coli-expressed F1 and G proteins. Western blot and IFA as well as ELISA showed that antisera against both protein had high titers with good reactivity and specificity. The present study has provided a base for development of diagnostic reagents for detection of NiV infection.

Cross-host evolution of severe acute respiratory syndrome coronavirus in palm civet and human.

The genomic sequences of severe acute respiratory syndrome coronaviruses from human and palm civet of the 2003/2004 outbreak in the city of Guangzhou, China, were nearly identical. Phylogenetic analysis suggested an independent viral invasion from animal to human in this new episode. Combining all existing data but excluding singletons, we identified 202 single-nucleotide variations. Among them, 17 are polymorphic in palm civets only. The ratio of nonsynonymous/synonymous nucleotide substitution in palm civets collected 1 yr apart from different geographic locations is very high, suggesting a rapid evolving process of viral proteins in civet as well, much like their adaptation in the human host in the early 2002-2003 epidemic. Major genetic variations in some critical genes, particularly the Spike gene, seemed essential for the transition from animal-to-human transmission to human-to-human transmission, which eventually caused the first severe acute respiratory syndrome outbreak of 2002/2003.

Civets are equally susceptible to experimental infection by two different severe acute respiratory syndrome coronavirus isolates.

Severe acute respiratory syndrome (SARS) was caused by a novel virus now known as SARS coronavirus (SARS-CoV). The discovery of SARS-CoV-like viruses in masked palm civets (Paguma larvata) raises the possibility that civets play a role in SARS-CoV transmission. To test the susceptibility of civets to experimental infection by different SARS-CoV isolates, 10 civets were inoculated with two human isolates of SARS-CoV, BJ01 (with a 29-nucleotide deletion) and GZ01 (without the 29-nucleotide deletion). All inoculated animals displayed clinical symptoms, such as fever, lethargy, and loss of aggressiveness, and the infection was confirmed by virus isolation, detection of viral genomic RNA, and serum-neutralizing antibodies. Our data show that civets were equally susceptible to SARS-CoV isolates GZ01 and BJ01.

[Genetic structure of Tujia as revealed by Y chromosomes].

Tujia people call themselves "Bizika", which means aboriginal. Genetic study of Tujia is virtually absent. To characterize the genetic structure of Tujia,the distribution of 14 Y haplogroups was studied in Tujia populations sampled from Enshi, Hubei (31 males) and Jishou, Hunan (68 males). A total of eight haplogroups were observed in the Enshi and Jishou populations. The haplogroup frequencies of Tujia were compared with the frequencies of other related ethnic groups, including Northern Han, Southern Han, Tibetan-Burman speaking populations,Daic and Hmong-Mien. The principal component (PC) analysis was conducted and the PCs were plotted to explore the historical migrations. In addition, partial correlation analysis was performed to study the relationship between the first three PCs and the haplogroups. The PC2 revealed a cluster of Tujia groups including Longshan,Yongshun, and Enshi with Lahu, suggesting possible interaction between Tujia and the Di-Qiang groups. However,a similarity between Han and Tujia populations, though differentiated, were also observed. We postulated, by incorporating the results of archaeological and historical evidences, that the Ba people, the ancestors of the Tujia,might be related with Di-Qiang groups and inhabited the Tujia area initially before a substantial interaction with Han and other ethnic groups.

Antibodies to SARS coronavirus in civets.

Using three different assays, we examined 103 serum samples collected from different civet farms and a market in China in June 2003 and January 2004. While civets on farms were largely free from SARS-CoV infection, approximately 80% of the animals from one animal market in Guangzhou contained significant levels of antibody to SARS-CoV, which suggests no widespread infection among civets resident on farms, and the infection of civets in the market might be associated with trading activities under the conditions of overcrowding and mixing of various animal species.