[Histone demethylase JMJD3 inhibits alveolar bone loss by regulating macrophage polarization in periodontitis].

Objective: To investigate the expression of histone demethylase, Jumonji domain-containing protein 3 (JMJD3), in inflammatory periodontal tissues and its potential mechanism for the regulation of periodontitis. Methods: The results of single-cell sequencing of periodontal tissues published in the Gene Expression Omnibus (GEO) database in 2022 were analyzed. Nine gingival samples each from healthy and inflamed periodontal patients were collected during periodontal surgery or tooth extractions for immunohistochemical staining and real-time fluorescence quantitative PCR (RT-qPCR). Mice periodontitis models were constructed, and the experimental groups were: healthy control+saline group, silk ligation+saline group, silk ligation+GSK-J4(inhibitor of JMJD3) group. Lipopolysaccharide (LPS) derived from Porphyromonas gingivalis (Pg) (Pg-LPS) was used to mimic the periodontal inflammatory microenvironment. The macrophages were treated with small interfering RNA (siRNA) targeting Jmjd3 and the JMJD3 inhibitor GSK-J4. siRNA transfection experiments were grouped into the following: the NC group (negative control sequence transfection group), the siRNA-Jmjd3 group, the NC+LPS group, siRNA-Jmjd3+LPS group. Inhibitor experiments were grouped as dimethyl sulfoxide (DMSO) group, GSK-J4 group, DMSO+LPS group, GSK-J4+LPS group. Western blotting and immunofluorescence staining were used to explore the effects of JMJD3 on macrophage polarization and periodontal inflammation in the in vivo and in vitro settings. Results: RT-qPCR results showed that JMJD3 expression in gingival tissues of periodontitis patients (1.97±0.91) was significantly higher than that in healthy gingival tissues (1.00±0.33) (t=2.45, P=0.048). RT-qPCR results of in vitro experiments showed that either siRNA knockdown of JMJD3 or inhibition of JMJD3 using GSK-J4 promoted M1 polarization and inhibited M2 polarization in macrophages under inflammatory environment: the expression of arginase I (Arg 1) in the NC+LPS group (0.90±0.06) was significantly higher than that in the siRNA-Jmjd3+LPS group (0.61±0.11) (P<0.01); the expression of interleukin (Il)-6, Il-1ß, and tumor necrosis factor alpha (Tnf-α) in the NC+LPS group (8.50±0.16, 5.56±0.20, 3.44±0.16) were significantly lower than those in the siRNA-Jmjd3+LPS group (14.63±0.48, 8.55±0.10, 11.72±0.16) (P<0.01). The expression of Arg-1, Ym1, Il-10 in the DMSO+LPS group (0.82±0.01, 0.35±0.16, 1.47±0.11) were significantly higher (P<0.01) than the GSK-J4+LPS group (0.55±0.03, 0.22±0.21, 0.51±0.11); the expression of Il-6, Il-1ß, and Tnf-α in the DMSO+LPS group (2.03±0.13, 3.63±0.14, 4.06±0.03) were significantly lower than the GSK-J4+LPS group (2.69±0.16, 15.04±1.15, 4.36±0.10) (P<0.01). The results of the in vivo experiments revealed that inhibition of JMJD3 exacerbated bone loss in experimental periodontitis mice, increased macrophage M1 polarization, and decreased M2 polarization in inflamed periodontal tissues. The buccal cemento-enamel junction (CEJ)-alveolar bone crest (ABC), palatal CEJ-ABC, as well as the ratio of M1/M2 type macrophages were significantly lower in the silk ligation+saline group [(0.26±0.03), (0.24±0.01) mm, 0.35±0.10) than in the silk ligation+GSK-J4 group [(0.34±0.04), (0.30±0.05) mm, 2.50±0.58) (t=3.65, P=0.006; t=2.67, P=0.049; t=7.31, P=0.004; respectively). Conclusions: Single-cell sequencing as well as the in vitro and in vivo experiments verified that JMJD3 expression was upregulated in periodontitis periodontal tissues. JMJD3 may exert a protective role in periodontitis by regulating macrophage polarization, thereby inhibiting alveolar bone destruction associated with the periodontitis.

[Research on the association between overweight and obesity mediated by Chinese children and adolescent aged 13-18 years physical exercise and psychological distress].

Objective: To analyze the association between overweight and obesity, psychological distress, and the influence of physical exercise among Chinese children and adolescents. Methods: The study used data from the 2019 Chinese National Survey on Students' Constitution and Health. A total of 95 280 Han Chinese children and adolescents aged 13 to 18 years were included in the study based on complete information on overweight and obesity, psychological distress assessment, and physical exercise variables. The Kessler Psychological Distress Scale was used to measure their psychological distress, with scores above 25 indicating high psychological distress. The BMI group of the adolescents was evaluated according to the Chinese standard "WS/T 586-2018 Screening for Overweight and Obesity in School-age Children and Adolescents", including underweight, normal weight, overweight, and obesity. The individual's BMI Z-score (BMIZ) was evaluated using the WHO standard and divided into ten groups based on the population percentile distribution. The survey collected the subjects' average daily physical exercise time and divided them into two groups: less than 1 hour and greater than or equal to 1 hour. Logistic regression analysis was used to analyze the relationship between overweight and obesity and high psychological distress among children and adolescents and the differences in association between different physical exercise groups. Results: The detection rates of overweight and obesity among Han Chinese children and adolescents aged 13 to 18 years in 2019 were 14.5%, and 7.6%, respectively. The rate of high psychological distress was 37.6%, and the rate of average daily physical exercise time exceeding 1 hour was 17.1%. Using the population with a BMIZ P10 below as a reference, the association strength between high psychological distress and the population gradually increased, with an OR (95%CI) value of 1.08 (1.02-1.14), 1.09 (1.02-1.14), 1.10 (1.03-1.16), and 1.16 (1.09-1.23) for BMIZ in the P60-, P70-, P80-, >P90 groups. Compared to normal weight, both overweight and obesity were positively associated with high psychological distress in children and adolescents, with obesity showing a more significant correlation, while underweight was negatively associated with high psychological distress. The detection rates of high psychological distress in normal weight, underweight, overweight, and obese groups were 37.6%, 37.0%, 38.2%, and 38.7%, respectively, with an OR (95%CI) value of 0.93 (0.88-0.98) for the underweight group and 1.05 (1.01-1.10) and 1.13 (1.07-1.19) for the overweight and obese groups, respectively. The positive correlation between BMIZ, overweight, and obesity with high psychological distress was significant only in adolescents who exercised less than 1 hour per day on average. Conclusions: Chinese children and adolescents face the problem of obesity and high psychological distress, with a positive correlation between these two problems. Physical activity may moderate the association between obesity and psychological distress in children and adolescents. Adequate physical exercise may offset the potential high psychological distress caused by obesity in children and adolescents. Thus, strengthening physical exercise among children and adolescents reduces the risks of both obesity and psychological health problems effectively.

[Research on the association between unhealthy lifestyle and psychological distress among Chinese children and adolescents aged 9-18 years].

Objective: To evaluate the level of psychological distress among Chinese children and adolescents and analyze its lifestyle influencing factors. Methods: Data were obtained from the 2019 Chinese National Survey on Students' Constitution and Health. A lotal of 120 285 Han Chinese children and adolescents aged 9-18 years with complete information on the psychological distress scale and lifestyle factors were selected, including 58 432 boys and 61 853 girls. The Kessler Psychological Distress Scale (K10) measured psychological distress, and lifestyles such as physical activity, sedentary behavior, diet, and sleep were also investigated. K10 scores of different genders were compared using the t-test, and the levels of psychological distress were compared using the χ2 test. Logistic regression was used to analyze lifestyle risk factors associated with high psychological distress, and multiple linear regression was used to find the relationship between K10 scores and lifestyle scores. Results: The average K10 score for Han Chinese children and adolescents aged 9-18 years was 21.25±7.35, with girls (21.43±7.35) scoring higher than boys (21.06±7.36), the difference was statistically significant (t=8.72, P<0.001). The rate of high psychological distress was 29.81%, with girls (31.08%) reporting higher rates than boys (28.46%), the difference was statistically significant (χ2=98.54,P<0.001). 56.10% of children and adolescents have unhealthy lifestyles, with girls (58.77%) reporting higher rates than boys (53.27%), the difference was statistically significant (χ2=368.53,P<0.001). Except for insufficient outdoor activities for girls (P=0.128), lifestyles such as insufficient physical activity, insufficient muscle-and-bone exercises, long screen time, not eating breakfast, eggs and dairy products every day, drinking sugary beverages once or more per day, and not having enough sleep are all risk factors for high psychological distress (all P<0.001). For every additional healthy lifestyle score, the K10 score decreased by 0.98 [ß=-0.98 (95%CI: -1.01- -0.95)] points (P<0.001). K10 scores in each region negatively correlate with lifestyle scores (all P<0.001). Among them, the K10 score in the eastern region showed the slightest decrease as the lifestyle score increased, while the western region showed the most decrease. Conclusions: The prevalence of psychological distress and unhealthy lifestyle in Chinese children and adolescents are high and interrelated. Compared those with healthy lifestyles, children and adolescents with unhealthy lifestyles are at greater risk of high psychological distress. Therefore, promoting healthy lifestyles for children and adolescents may be one of the important ways to improve their mental health.

[Research on the association between ambient PM2.5 and its components and psychological distress among Chinese children and adolescents aged 9-18 years].

Objective: To analyze the association between exposure to ambient PM2.5 and its components [sulfate (SO42-), nitrate (NO3-), ammonium (NH4+), organic matter (OM), and black carbon (BC)] and psychological distress among school children and adolescents aged 9 to 18 years in China. Methods: Based on data from the 2019 Chinese National Survey on Students' Constitution and Health, 130 808 children and adolescents aged 9 to 18 years were included. Scoring and determination of higher psychological distress were based on the Kessler Psychological Distress Scale (K10), and annual average exposure data for air pollution were matched by survey location and time. We used a restricted cubic spline approach based on logistic regression to fit the associations between PM2.5, SO42-, NO3-, NH4+, organic matter, black carbon concentrations, and psychological distress. Logistic regression models were used to analyse different age, gender, BMI and physical activity subgroups to estimate the association between exposure to high levels of pollution and high levels of psychological distress in different subgroups. Results: The proportion of children and adolescents with high levels of psychological distress in China was 30.4%, with girls being higher than boys at 31.6% and 29.1%, respectively (P<0.05). A non-linear positive association existed between exposure to PM2.5, SO42-, NO3- and organic matter concentrations and higher psychological distress. As PM2.5, NO3-, NH4+, organic matter, and black carbon concentrations continued to rise, the increase in the risk of higher psychological distress slowed, while SO42- showed little change in the OR of psychological distress at lower concentrations but continued to rise at higher concentrations. PM2.5 and its components were statistically associated with psychological distress in the physically inactive group but not in the physically active group. The association between high levels of pollutants and high psychological distress was stronger among students aged 9 to 12 years compared with students aged 13 to 15 years and 16 to 18 years. Conclusions: The ambient pollutant PM2.5 and its components are associated with psychological distress in children and adolescents aged 9 to 18 years in China. High pollutant exposure is a risk factor for high psychological distress among physically inactive children and adolescents, and there are age differences in the association between PM2.5 and components and psychological distress.

[Study on 4 cases of mushroom poisoning with amanita and identification of poison].

Different kinds of poisonous mushrooms contain different toxic components. Acute liver injury caused by amanita mushroom is the main cause of death from poisonous mushroom poisoning in China. Consumption of poisonous mushrooms has an incubation period, there is a false recovery period in the clinical process, and the early performance is slight and does not attract enough attention from doctors, and it is easy to miss the treatment opportunity. The clinical characteristics, treatment and identification of mushrooms containing amanita in 4 patients were analyzed in order to improve clinicians' understanding of the diagnosis and treatment of mushroom poisoning and early species identification.

[Progress on specific and non-specific immune effects of Bacille Calmette-Guerin].

Currently, Bacille Calmette-Guerin(BCG) is still the only admitted vaccine to prevent tuberculosis around the world. The target population is infants and children, but its protective efficacy is limited. As more and more studies have shown that re-vaccination with BCG protects against tuberculosis in adults, BCG can also induce non-specific immunity against other respiratory diseases and some chronic diseases by training immunity, especially the immune effects against COVID-19. At present, the epidemic of COVID-19 has not been effectively contained, and it is worth considering whether BCG vaccine can be used as an intervention to prevent COVID-19. The WHO and China do not have a policy to support BCG revaccination, and as more and more BCG vaccines are discovered, whether selective revaccination can be carried out in some high-risk populations and whether the vaccine can be used more widely have led to intense discussions. This article reviewed the effects of specific immunity and non-specific immunity of BCG on tuberculosis and non-tuberculous diseases.

[Long-term outcomes of Bryan artificial cervical disc replacement for degenerative cervical spondylosis].

Objective: To compare the long-term clinical effect and imaging results of Bryan artificial cervical disc replacement (ACDR) and anterior cervical discectomy and fusion (ACDF) and to explore whether ACDR can reduce the occurrence of postoperative adjacent segment degeneration (ASD) in the treatment of degenerative cervical spondylosis. Methods: It was a retrospective study. Total of 60 patients with degenerative cervical spondylosis, who had received operations by Bryan ACDR (n=27) and ACDF (n=33) in the Third Hospital of Hebei Medical University between January 2005 and December 2009 were enrolled in this study. The Japanese Orthopedic Association (JOA) scores, neck disability index (NDI), visual analogue scale (VAS), Odom's scale, cervical range of motion (ROM), surgical segment ROM, heterotopic ossification (HO) and ASD were used to evaluate the clinical and radiologic results. The indices abovementioned were compared between the two groups. Results: The VAS, NDI and JOA scores at each follow-up node in both ACDR and ACDF group were all significantly improved when compared with those before operation (all P<0.05), but there was no significant differences between the two groups (all P>0.05). Of all, 88.9%(24/27) of patients in ACDR group and 84.8%(28/33) of patients in ACDF group achieved excellent or good results, however, there was no significant difference in Odom's scale between the two groups (P>0.05). At the last follow-up, the cervical ROM in ACDR group was 43.2°±8.8°, which was slightly lower than that before operation (45.7°±10.4°), the difference was not statistically significant (t=0.954, P=0.345); the surgical segment ROM in ACDR group was 5.9°±3.6°, which was significantly reduced when compared with that before operation (8.8°±3.4°, t=3.043, P<0.01). However at the last follow-up, the cervical ROM in ACDF group was 36.4°±8.4°, which was significantly reduced when compared with that before operation (43.9°±11.1°), the difference was statistically significant (t=3.095, P<0.01). Although, there was no significant difference in the cervical ROM between the two groups before operation (t=0.643, P=0.523), and the difference was statistically significant at the last follow-up (t=3.054, P<0.01). At the last follow-up, the incidence of HO in ACDR group was 92.6%, and the high-grade HO was 37.0%. The incidence of ASD in ACDR group was 39.5%, which was much lower than that in ACDF group (61.1%, χ(2)=4.462, P=0.035). Conclusion: At minimum follow-up of 10 years, Bryan ACDR achieves a satisfactory clinical effect consistent with ACDF. In terms of advantages, ACDR could maintain the ROM of cervical and retain the ROM of the surgical segment, which reduces the occurrence of ASD by preserving motion.

[Surveillance on the pathogen of imported Dengue fever in Henan province, 2018].

Objective: To analyze data gathered from the laboratory records related to imported Dengue cases in Henan province in 2018. Methods: Suspected Dengue cases were found out through the Dengue fever surveillance network from the National infectious Disease reporting management information system. Serum samples of suspected Dengue cases were collected while case study and tested for Dengue NS1 antigen, IgM, IgG antibodies and Dengue RNA in Henan province in 2018. According to the standardized Dengue diagnosis criteria, confirmed cases were identified under the results of testing. Dengue RNA was checked by Real-time PCR genotyping and amplification of E gene in the samples being tested, before the PCR products were sequenced and analyses of homological and phylogenetic were performed. Results: In 2018, a total of 29 cases of Dengue fever was reported in Henan province, with all of which were imported cases, mainly from Southeast Asian countries and Africa. Majority of the cases were young and middle-aged farmers under 45 years old, and the number of males was significantly higher than that of females. The imported cases were dispersed in time and space. Among the 29 Dengue reported cases, 22 cases were with NS1 antigen and/or IgM positive through testing, while 6 cases were positive by detection of the Dengue virus RNA. These 6 samples with Dengue RNA were genotyped successfully, including 3 cases of Dengue virus type 1 and 3 cases of type 2. One of the Maldives import Dengue virus type 2 samples was sequenced. Result showed that the sequence belonged to the Asian Ⅰ genotype, which was most consistently similar to the Cambodia's Dengue virus type 2 JF730046, identified in 2008. Conclusions: The incidence of imported Dengue fever cases increased significantly in Henan province in 2018, compared to that in 2017, but fortunately did not cause any local epidemics.

[Analysis on the epidemiology and etiology characteristics of first imported Chikungunya fever case in Henan Province in 2017].

To study the epidemiology and etiology characteristics of first imported Chikungunya fever case in Henan province, China, 2017. The patient was confirmed by Chikungunya virus (CHIKV) infected as CHIKV ribonucleotide was continuously detected in his serum specimens. BHK-21 cell line was used for virus isolation, the strain was named CHIKV/Henan001/2017. CHIKV/Henan001/2017 belonged to genotype ECSA. The highest ribonucleotide homology sequence of highly conserved region E1 with CHIKV/Henan001/2017 was hk02 strain (99.8%), who was an imported strain to Hong Kong, China, 2016. Epidemiological information and laboratory testing confirmed it was an imported Chikungunya fever case in Henan province, 2017. No secondary case has been reported.

[Laboratory diagnosis and molecular tracing of dengue bordline cases in Henan Province, 2017].

Objective: To confirm the laboratory diagnosis of dengue bordline cases reported in Henan Province and trace its origin from molecular level in 2017. Methods: The study samples were blood samples (3-5 ml), which came from 8 suspected cases of dengue fever reported in the 2017 direct reporting system of Henan provincial infectious disease monitoring network. Meanwhile, case investigation was conducted according to National dengue fever surveillance programme. Serum were separated from blood samples and tested for Dengue NS1 antigen, IgM & IgG antibodies, and dengue RNA. According to dengue diagnosis criteria, confirmed cases were identified by testing results. Samples carried dengue RNA performed for real-time PCR genotyping and amplification of E gene. Then, the amplicons were sequenced and homological and phylogenetic analyses were constructed. Results: 8 serum samples of suspected dengue cases were collected in Henan Province, 2017. Six of them were diagnosed as dengue confirmed cases. All the dengue confirmed cases belonged to outside imported cases, 5 of them were positive by dengue RNA testing. Genotyping results showed there were 1 DENV1 case, 2 DENV2 cases and 2 DENV3 cases. A DENV2 case and a DENV3 case of this study were traced its origin successfully. The sequence of Pakistan imported DENV2 case belongs to cosmopolitan genotype, which was the most consistent with Pakistan's DENV2 KJ010186 in 2013 (identity 99.0%). The sequence of Malaysia imported DENV3 case belongs to genotype I, which was the most consistent with Singapore's DENV3 KX224276 in 2014(identity 99.0%). Conclusion: The laboratory diagnosis and molecular traceability of dengue cases in Henan Province in 2017 confirmed that all cases were imported and did not cause local epidemics.

[Preliminary study of robot-assisted ankle rehabilitation for children with cerebral palsy].

OBJECTIVE: To propose a kind of robotic ankle-foot rehabilitation system for children with cerebral palsy and to preliminarily verify its feasibility in clinical application. METHODS: A robot assisted ankle-foot rehabilitation system was specially designed and developed for children with cerebral palsy and a preliminary clinical study was conducted in Department of Rehabilitation Medicine, Peking University First Hospital. Modified Tardieu Scale and joint biomechanical properties (ankle plantar flexion resistance torque under different ankle dorsiflexion angles) were measured to analyze the muscle tone and soft tissue compliance of the ankle plantar flexors pre- and post-robotic training intervention. Six children with cerebral palsy (4 girls and 2 boys, mean age: 7 years) were recruited in this study. Each subject received 5 session robotic training and each session included 10-cycle passive stretching and static hold. SPSS 19.0 software was used for data statistical analysis. RESULTS: Both R1 and R2 angles of Modified Tardieu Scale for ankle plantar flexors after training were significantly higher than those before the treatments (Gastrocnemius: PR1=0.003, PR2=0.029; Soleus: PR1=0.002, PR2=0.034). The difference between R2 and R1 was of no statistical difference before and after the training (P=0.067 and P=0.067, respectively). After training, the ankle plantar flexion resistance torque under different dorsiflexion angles (0°, 10°, 20°, 30°) were significantly reduced than those before training (P=0.001, P=0.001, P=0.014, P=0.002, respectively). CONCLUSION: The robot assisted ankle-foot rehabilitation system can improve the contracture and soft tissue compliance of cerebral palsy children's ankle plantar flexors. All the children in the study were well tolerated and interested with the training, easy to accept and cooperate with it. This device may be suitable for application in the rehabilitation of children with cerebral palsy. However, further randomized clinical trials with larger sample size are still needed to verify the long term efficacy of this device.

Changes in the gut microbiome of the Chinese mitten crab (Eriocheir sinensis) in response to White spot syndrome virus (WSSV) infection.

Intestinal microorganisms play important roles in maintaining host health, but their functions in aquatic animal hosts have yet to be fully elucidated. The Chinese mitten crab, Eriocheir sinensis, is one such example. We attempted to identify the shift of gut microbiota that occurred in response to infection of white spot syndrome virus (WSSV), an emerging viral pathogen in the crab aquaculture industry. The microbiota may exert some control over aspects of the viral pathogenesis. We investigated the changes in composition and structure of the crab gut microbiome during various WSSV infection stages of 6 h post-infection (hpi) and 48 hpi, using a 16S rRNA approach on the MiSeq Illumina sequencing platform. Four phyla (Firmicutes, Proteobacteria, Tenericutes and Bacteroidetes) were most dominant in the gut of E. sinensis regardless of the WSSV infection stages. However, further analysis revealed that over 12 bacterial phyla, 44 orders and 68 families were significantly different in abundance at various states of WSSV infection. Several intriguing aspects of E. sinensis gut bacteria that had not been previously reported were also uncovered, such as class Mollicutes was dominant here, but absent in crabs from Yangtze River estuary and Chongming Islands. Overall, this study provided the first evidence that changes in gut microbiome were closely associated with the severity of WSSV infection and that indicator taxa could be used to evaluate the crab health status.

[Surface roughness and gloss of novel flowable composites after polishing and simulated brushing wear].

Objective: To investigate surface properties of novel flowable composites after polishing and simulated brushing wear, compared to their pasty counterpart. Methods: Composites employed in this study were: three flowable composites (A1: Clearfil Majesty ES Flow; B1: Beautifil Flow Plus F00; C1: Filtek Bulk Fill) and three paste composites (A2: Clearfil Majesty; B2: Beautifil; C2: Filtek Z350. Eleven disk-shaped specimens were made for each material. The specimens were cured, then subjected to sandpaper finishing for 20 s, one-step polishing for 30 s, finally subjected to simulated brushing for 10 000 cycles. Surface roughness and glossiness were measured before finishing, after finishing, after polishing, after 5 000 brushing cycles and after 10 000 brushing cycles, respectively. Data obtained were analyzed using two-way ANOVA method. Scanning electron microscope was employed to examine the microscopic appearance of each material. Results: Surface roughness (0.11~0.22 µm) and glossiness (74.25~86.48 GU) of each material were similar after one-step polishing. After brushing simulation, roughness increased significantly and glossiness decreased significantly for each material (P<0.05). Group A1 presented the best gloss ([50.68±1.58] GU) after final wear (P<0.05). Flowable composites of group A1 and B1 tested in the present setup showed better surface properties compared to their pasty counterpart (group A2 and B2). Conclusions: Within the limit of this study, flowable composites tested in the present research can obtain similar surface polish or even better than the paste composite counterpart.

Nutritive value and the maximum inclusion level of pennycress meal for broiler chickens.

Two experiments were conducted to evaluate the nutritive value and maximum safe level (MSL) of pennycress meal (PM) for broiler chicks. In experiment 1, a total of 480 chicks was fed either mash or crumbled diets containing zero, 5, 10, or 15% PM for 18 d (8 diets; 6 replications per diet). In experiment 2, a total of 660 chicks was fed mash diets containing zero, 3, 6, 9, 12, or 15% of either PM or canola meal (CM; a comparative reference) for 14 d (11 diets; 6 replications per diet). Analytical results show that PM is a good source of protein (∼31% CP) and it is very comparable to CM (∼36% CP). However, it contains higher erucic acid (∼1.68 vs. < 0.021%), glucosinolates (sinigrin) (∼63.5 vs. <0.163 µmol /g), and crude fiber (18.60 vs. 9.27%) compared to CM. In experiment 1, increasing PM from zero to 15% resulted in linear reductions (P < 0.05) in FI, BWG, and FCR at 10 days. Above 10%, performance responses were affected for FI and BWG at 18 d, respectively. An estimated MSL of 10% PM based on orthogonal contrast was optimal for satisfactory FI and BWG. The MSL as estimated by broken-line linear (BLL) and broken-line quadratic (BLQ) models was 9.12 ± 0.50 and 7.0 ± 1.27%, respectively. In experiment 2, growth performance at 14 d was reduced above 9% due to PM inclusion. CM inclusion did not affect growth performance at 14 d, suggesting 15% to be safe. The MSL for maximum growth performance varied depending on the statistical analysis as follows: 12% by orthogonal contrast and LSD, 15% by the Scheffé test, 10.84 ± 0.57 by BLL, and 8.61 ± 1.29 by BLQ. In conclusion, PM can be included in broiler starter diets as a protein source but its inclusion should be limited to no more than 8.5%. Different statistical procedures give different MSL and this should be considered when interpreting the data.

Development of In situ hybridization and real-time PCR assays for the detection of Hepatospora eriocheir, a microsporidian pathogen in the Chinese mitten crab Eriocheir sinensis.

A microsporidian parasite, Hepatospora eriocheir, is an emerging pathogen for the Chinese mitten crab Eriocheir sinensis. Currently, there is scant information about the way it transmits infection in the crustacean of commercial importance, including its pathogenesis, propagation and infection route in vivo. In this study, chromogenic in situ hybridization (ISH) and quantitative real-time PCR (qPCR) assays were developed to address this pressing need, and we provided an advance in the detection methods available. Pathogens can be seen in situ with associated lesions using ISH. Positive hybridization signals were noted inside the epithelial cells of the hepatopancreas, and putative free parasite spores were observed within the tubule lumen, which were associated with lesions detected by electron microscopy and haematoxylin and eosin (H&E) analysis. qPCR allows the determination of parasite loads in infected tissues, which is important for understanding disease progression and transmission. The hepatopancreas displayed the biggest statistical copy numbers among different tissues of infected crabs, confirming a tissue-specific pathogen infection characteristic. The qPCR assay also proved to be suitable for the diagnosis of asymptomatic carrier crabs. Combination of the two methods could facilitate the study of H. eriocheir infection mechanism in E. sinensis, enhance the early diagnosis of the pathogen and improve the management of microsporidian diseases in commercial crustaceans.

[The comparison of clinical features and laboratory indexes between flat descending hearing loss and total hearing loss].

Objective:To discuss similarities and differences in clinical features and laboratory indexes between patients with flat descending type sudden hearing loss and those with total hearing loss. Method:The clinical data of 123 patients with full frequencies hearing loss were retrospectively analyzed. The differences in clinical features and laboratory tests(platelet, coagulation series, D-dimer, blood lipids, hemorheology) between patients with flat descending hearing loss and those with total hearing loss were analyzed by gender, age and ear side, treatment time, concomitant symptom (tinnitus, dizziness), original underlying diseases (hypertension, diabetes), etc. Result:In the clinical features,among 51 flat descending cases, the ratio of male and female was 2.401:1; among 72 total hearing loss cases, the ratio of men and women ratio was 1.058:1 (P<0.05). Among two groups of patients,the majority received treatment within 7 days, among whom 66.7% were flat descending population, and 83.3% were total hearing loss population (P<0.05). Flat descending population with dizziness only accounted for 35.3% while this figure was up to 70.8% when it came to total hearing loss patients (P<0.01). Two groups showed no differences in age, ear side, tinnitus, the original underlying diseases (hypertension, diabetes). In the laboratory tests, the total hearing loss population overtopped the plat descending population in PLT and PCT (P<0.05), while falling below the plat descending population in APTT (P<0.01). Two groups showed no differences in other indicators of platelet and coagulation series and laboratory data of D-dimer, blood lipids, hemorheology. Conclusion:Compared with flat descending sudden hearing loss, sudden total hearing loss more frequently happened to females who also were accompanied by dizziness. The treatment rate within 7 days was high and the patients with hypercoagulable state accounted for a higher proportion.

Molecular evolutionary analysis of the high-affinity K+ transporter gene family in angiosperms.

The high-affinity K(+) transporter (HKT) family comprises a group of multifunctional cation transporters widely distributed in organisms ranging from Bacteria to Eukarya. In angiosperms, the HKT family consists primarily of nine types, whose evolutionary relationships are not fully understood. The available sequences from 31 plant species were used to perform a comprehensive evolutionary analysis, including an examination of selection pressure and estimating phylogenetic tree and gene duplication events. Our results show that a gene duplication in the HKT1;5/HKT1;4 cluster might have led to the divergence of the HKT1;5 and HKT1;4 subfamilies. Additionally, maximum likelihood analysis revealed that the HKT family has undergone a strong purifying selection. An analysis of the amino acids provided strong statistical evidence for a functional divergence between subfamilies 1 and 2. Our study was the first to provide evidence of this functional divergence between these two subfamilies. Analysis of co-evolution in HKT identified 25 co-evolved groups. These findings expanded our understanding of the evolutionary mechanisms driving functional diversification of HKT proteins.