Foeniculum vulgare Mill., commonly called fennel, is a medicinal plant belonging to the Umbelliferae (Apiaceae) family, and is used in traditional medicine. Antibacterial peptides were isolated using sodium phosphate citrate buffer and, for extraction, cetyltrimethyl ammonium bromide (CTAB) buffer with pH 6, have been employed and antimicrobial activity tested against four reference strains. The extracted protein was subjected to 3 kDa dialysis and separation was carried out by DEAE-ion exchange chromatography and further proteins were identified by 2D gel electrophoresis. The results of Foeniculum vulgare elutes obtained from DEAE-ion exchange chromatography were tested for antibacterial activity. Elute 3 shows the highest antibacterial activity on Pseudomonas aeruginosa with a diameter of a zone of inhibition of 16 mm and IC50 value 25.02 (mcg/mL). Based on the findings of the wide usage in treatment of various ailments and day-to-day life, Foeniculum vulgare seeds were used in the present research and have shown promising antibacterial effects, which requires further proteomic research to authenticate the role of the anticipated proteins. Voluminous plants and plant extracts have a wide variety of resistance mechanisms to chemical, physical, and biological strains, like heavy metals, cold, drought, pathogens and contaminant attacks from bacteria, fungi, and viruses. Several plants exhibited complete genes, allied together for assimilated control over the infections from numerous types of pathogens . TypicalAntimicrobial peptides are pervasive and the established host resistances, in contrast to pathogens and pests in different organisms, vary from microorganisms to animals . AntimicFoeniculum vulgare Mill., commonly called fennel, is a therapeutic plant belonging to the Umbelliferae (Apiaceae) family, is widely used in traditional medicine. This is one the most important plants, which has several pharmacological properties, both in vivo and in vitro, including anti-microbial, anti-viral, anti-inflammatory, anti-mutagenic activities, etc. [Foeniculum vulgare emerged as a good source of medicinal products for research, proving noteworthy in the field of pharmaceutical biology, as well as in the research and development for new drugs. Previous study showed that extract from Foeniculum vulgare has significant anti-bacterial activity against some tested foodborne pathogens [es, etc. . Based oathogens . There aathogens ,11, defeathogens ,13, thioathogens , lipid tathogens ,16, 2S aathogens ,18, and athogens ,20,21. Sathogens ,22.Identification and characterization of proteins is an essential step to understand the key roles of proteins in the cell. In recent times, proteomic analysis has emerged as a successful technology to identify and characterize the loop link proteins that will merge the gap between proteomics and pharmacology. In our study we adopted the gel electrophoresis technique to separate proteins by 2DE, followed by protein identification by mass spectrometry, which is a widely used approach in proteomics.Escherichia Coli (E. coli), Pseudomonas aeruginosa (P. aeruginosa), Staphylococcus aureus (S. aureus), and Proteus vulgaris (P. vulgaris), and compared with the standard antibiotics ciprofloxacin (25 mcg/mL) and chloramphenicol (100 mcg/mL).The protein extract after extracting in sodium acetate buffer, pH-6.5, were purified by dialysis, and the same extract was concentrated and subjected for antibacterial testing against four standard pathogenic bacterial strains found to cause foodborne illness and spoilage of food and herbal drugs. The bacterial strains used in the study were Foeniculum vulgare Mill., showed low concentration of protein, at 80 μg/mL, when extracted in sodium phosphate citrate buffer (pH-7.2) and CTAB buffer (pH 6.0), and exhibited the highest concentration of protein, at 140 μg/mL, with sodium acetate buffer (pH-6.5).The result of the study shown in Foeniculum vulgare Mill. (S1) were found to exhibit nil activity on all of the bacterial strains used, so this plant’s seeds were again extracted in sodium acetate buffer, pH-6.5, to obtain a sensitivity pattern on the bacterial strains. Pseudomonas aeruginosa exhibited very good sensitivity when compared with the other bacterial strains with a diameter of the zone of inhibition 12.5 mm, whereas Staphylococcus aureus and Proteus vulgaris showed similar zones of inhibition of 12 mm. E. coli, Pseudomonas aeruginosa, and Proteus vulgaris revealed good zone of inhibition patterns of 11 mm, 12.5 mm, and 12 mm, when compared with the standard antibiotic Chloramphenicol (25 mcg/mL), which is only 8 mm.Antibacterial activity results shown in Foeniculum vulgare Mill. extracted in sodium acetate buffer at a pH of 6.5 after dialysis were further purified using the DEAE-ion exchange chromatography technique to isolate and characterize the antibacterial proteins present in the extract. The crude protein extract after dialysis was subjected to ion exchange chromatography, four elutes were obtained, and the concentration was found using the Lowry method. The protein concentration was found in ranges from 100–120 mcg/mL, the highest concentration was seen in elute 1 and 2 with 120 mcg/mL, and the least was found in elute 4, with 100 mcg/mL, results shown in the The crude protein extracts from Foeniculum vulgare elutes obtained after ion exchange chromatography were tested for antibacterial activity as shown in P. aeruginosa with a diameter of the zone of inhibition of 16 mm and an IC50 value of 25.02 mcg/mL, when compared with the standard antibiotic Chloramphenicol at 100 mcg/mL which is only 12 mm and has an IC50 value of 14.634 mcg/mL. Elute 2 shows a good zone of inhibition on Proteus vulgaris with a diameter of the zone of inhibition of 13 mm and an IC50 value of 57.83 mcg/mL, and the lowest zone of inhibition was seen on P. aeruginosa with a zone of inhibition of 4 mm and an IC50 value of 68.33 mcg/mL. S. aureus was found to shown good sensitivity toward elute 4 with a diameter of the zone of inhibition of 12 mm and an IC50 value of 20.8 mcg/m) and, with elute 1, a diameter of the zone of inhibition of 10 mm and an IC50 value 27.64 mcg/mL. E. coli exhibited good sensitivity towards elute 1 with a zone of inhibition at 13 mm and IC50 value of 67.56 mcg/mL.The results of 50 of different elutes on four bacterial strains used in the study. Among the tested strains, S. aureus and P. vulgaris showed the highest significant inhibition in comparison to standard cipro (p = 0.02) whereas there is no significance observed in the IC50 in E. coli and P. aeruginosa compared to standard cipro, results shown in The analysis was performed using SAS for Windows version 9.2 . One-way ANOVA was used to compare the ICFoeniculum vulgare seeds after anion exchange chromatography were subjected to 2-DE. Out of four elutes obtained from anion exchange chromatography, only elute 2 was used for 2-DE analysis. The elute 2 protein sample extracted from Foeniculum vulgare seeds was analyzed to investigate the proteins present in the sample based on the molecular weight, using HP Scanjet G4010 proteomic techniques. The analysis of 2D gel of Foeniculum vulgare showed reproducible protein spots, most of which were distributed near the center of the gels, between pI values 5–8 and molecular weight of 34.4 kDa and 48 kDa . Past studies have demonstrated that asymptomatic bacteriuria posed more serious hazards in pregnant women [Antibiotic resistance is one the most dangerous global problems today, which makes antimicrobial proteins/peptides the agents of a substitute for cures of pathogenic diseases . In thisnt women , so bactnt women ,26.In the present research work on anion exchange resin diethyl amino ethyl cellulose (DEAE) was used, which is positively-charged to exchange negatively-charged protein. DEAE-ion exchange chromatography is one of the promising techniques used to separate proteins based on their surface ionic charge using resins that are altered with either negatively-charged or positively-charged biochemical groups . As in tFoeniculum vulgare seed extract, and most of the protein spots were found to be between pI values 5–8 and molecular weights of 34.4 kDa and 48 kDa on the 2D gel, which was found to be higher in molecular weight and possess antibacterial activity from being tested on the four bacterial strains. Previous study identified napins, a class of proteins from 2S albumin, which were isolated from plants and that can be easily extracted from seeds [Proteomics is a handy technique used in the characterization of various proteins . Additioom seeds  which arom seeds . Furtherom seeds  and coliom seeds ,34,35, aFoeniculum vulgare Mill. seeds were procured from a local market in Riyadh, Saudi Arabia. Before adopting the protocol, the plant seeds were identified by Dr. Ali S. Alqahtani, Pharmacognosy Department, King Saud University, Riyadh, Saudi Arabia. Ethical approval was obtained from the research ethics committee (CAMS-153-36/37) from the college of Applied Medical Sciences, King Saud University, Riyadh, Saudi Arabia.The Foeniculum vulgare Mill. seeds were cleaned using sterile water to remove any debris left over on the seeds and dried in a biological safety cabin to maintain the sterile condition. Later, fine powder was prepared from the seeds under aseptic conditions and the extract was soaked in sodium phosphate buffer (pH-6.5) at 30 °C and left for overnight incubation. The next day, extracts were filtered with Whattmann filter paper No. 1 and subjected to 80% ammonium sulfate saturation. The collected saturated material was then separated by dialysis using a 3 kDa cutoff dialysis tubing  and the protein’ concentration was further estimated by spectroscopic analysis at a wavelength of 280 nm.The The chromatography column was prepared using a DEAE cellulose bed of 1 cm thickness, washed with ethanol before use, 20 mM Tris-Hcl for the 250 mL required volume was prepared, and pH was attuned to 8.5 using NaOH. Bed preparation: counter ions  of 40 mL of 25 mM NaCl in 50 mM tris (pH-7.2) and 0.3% of DEAE were applied to the column. The elutes were run for 4 h and 2 mL was collected. Each column used a flash chromatography column from Sigma Aldrich, St. Louis, , and the eluting buffer was sodium phosphate citrate buffer (pH-7.2). Elute preparation: four elutes were prepared in test tubes for each dialyzed sample. After dialysis, the samples were poured into the column without disturbing the bed, and left for 20 min to settle. The first eluting buffer, i.e., sodium phosphate citrate buffer (pH-7.2), was loaded into the column and was used to elute the sample. Thed column was allowed to settle for 15–20 min. The flow rate is adjusted to 1 mL/min and the elute was collected in test tubes at the bottom.E. coli, P. aeruginosa S. aureus, and P. vulgaris, and were obtained from department of laboratory sciences, College of Applied Medical Sciences, Riyadh, Saudi Arabia.Bacterial strains used for the study were Antibacterial activity of this crude extract was tested against these four strains in Hi-media. Culture slants were prepared by taking a loop full of pure culture and dispensing into 10 mL nutrient broth from four strains in a sterilized laminar flow and incubated for two days in the incubator. The final concentration of each active culture was maintain at 108 cfu/mL.The antibacterial activity of the crude protein extracts was determined by agar well diffusion assay . Pure cu8 cfu/mL concentration added to each well and made a final volume to 200 μL by adding LB broth. Ten microliters of MTT (5 mg/mL) was added to all of the wells after incubation at ±4 °C for 18 h and the readings were taken at 530 nm using an ELISA reader . The minimum concentration of protein that showed total inhibition was considered as its MIC minimum zone of inhibition. As control experiments, Chloramphenicol (25 mcg) and Ciprofloxacin (100 mcg) were taken as positive controls, and sterile broth as the negative control.The protein extracts of different plant seeds were tested for antibacterial activity against four strains by the micro-dilution method , with so50) was the parameter used to compare the antibacterial activity. A lower IC50 means better antibacterial activity. All of the tests were performed in triplicate and the results were expressed as the mean ± standard deviation. The percent of inhibition of antibacterial activity of the protein was calculated based on: % Inhibition = (A control − A sample)/A control × 100.The inhibition concentration at 50% inhibition  electrophoresis, one the most important tool in proteomics, was performed in accordance with the method reported in . The samFoeniculum vulgare seeds have shown promise in traditional medicine for the treatment of a wide range of diseases. From our results, 14 spots from the 2D gel were eluted and further subjected to mass spectrometry studies, undergoing more proteomic analyses and the isolation of small peptides from Foeniculum vulgare. Many studies have reported that this plant has numerous in vivo and in vitro pharmacological assets that can be used in various aspects of prognosis. Based on the findings of the wide usage in treatment of various ailments and day-to-day life, Foeniculum vulgare seeds were used in the present research and have established the promising antibacterial effects of these seeds, which require further proteomic research to authenticate the role of anticipated proteins.Over a long period of time,


N = 532) reported values using the Portrait Values Questionnaire and rated their postulated behaviors with the Everyday Behavior Questionnaire. A confirmatory factor analysis validated the discriminant validity of 19 refined values. Multidimensional scaling analyses revealed a circular motivational structure with national characteristics. More importantly, the correlation analysis showed that 13 of 19 values correlated most strongly with its postulated or adjacent behaviors, which supported the cross-cultural prediction of behaviors by values. Variations differing from theoretical structure and the moderating of value-behavior relations by gender better reflected the effects of Chinese traditional culture, such as harmony, benevolence, and “the Golden Mean.” The results of this study enrich the cross-cultural validity of the refined value theory among Chinese minorities and enhanced our understanding of the multiculturally influenced minority population.The minorities in Southwest China are characterized by a blend of diverse cultures. It is not clear how values predict behaviors in such a population. We applied Schwartz's refined value theory to assess the association between values and behaviors. Respondents ( Associations of values with behaviors have frequently been the focus of attention in psychological studies. Many researchers have studied values on the assumption that they can explain, influence, and even predict behavior in hypothetical   Geary,  and the ) Geary, , the gro) Geary, .We carried out our studies in a multicultural but small population, which has been seldom included in previous publications. The minorities were from Southwest China, mainly including the province of Yunnan, Guizhou, Sichuan, and Chongqing, and comprised more than 30 of the 55 official Chinese minority groups. Each group either inhabits a specific region in Southwest China, or is dispersed among other ethnicities. Despite such high centralized and multicultural minority distribution, Southwest minorities manifested more harmony, and intercultural fusion Zheng, . Such a In this study, we followed the same procedure and analyses conducted by Schwartz and Butenko  to validn = 393) were students, and the other 26.1% (n = 139) included teachers, workers, peasants, freelancers, physicians and nurses. In the sample, 53.6% (n = 285) were women, 71.4% (n = 379) were aged 17–24 years, 18.4% (n = 98) were aged 25–35 years, 9.8% (n = 52) were aged 36–49 years, 91.9% (n = 489) were undergraduates or with higher educational levels, 91.4% (n = 486) were non-religious, 81.8% (n = 435) were unmarried, 83.8% (n = 83.8) were skilled Mandarin masters, 62.4% (n = 332) were descendants of the same ethnic minority group, and 69.7% (n = 371) were from the minority-inhabited area.The final analysis was conducted on 532 surveys in Chongqing, Yunnan, Guizhou, and Sichuan provinces in China. Respondents included members from 31 of the 55 official ethnic minorities in China including Yi, Miao, Bai, Tujia, Hui, Tung, Zhuang, Hani, Dai, Bouyei, Gelo, Nu, Lisu, Jingpo, Shui, Naxi, Tibetan, Wa, Achang, Blang, Jinuo, Qiang, Yao, Lahu, Dongxiang, Pumi, Drung, Manchu, She, Li, and Maonan. Occupationally, 73.9%  or to their minority acquaintances  to fill out. Additionally, two teachers from Southwest Minzu University delivered our questionnaires to their minority students during a class session. The completed questionnaires were collected on the spot within about 30–45 min. The participants' ethnic minority identification was self-reported, and we confirmed their identification through telephone fellow-up. The participants got a monetary reward (10 Yuan RMB) after completing two self-report questionnaires as a token of our gratitude for their participation.not like me at all) to 6 (very much like me). For example, “Having ambitions in life is important to him” describes a person who emphasizes achievement values; “Being wealthy is important to him” describes a person who values power highly; “Obeying all the laws is important to him” describes a person who thinks that humility is important. The 57 items could be combined into 10 basic values or 19 narrowly refined values. The internal reliability of the 10 values subscales ranged from 0.631 to 0.825 and the 19 values subscales ranged from 0.606 to 0.816 in this study.”We assessed values with a Chinese Simplified Female and Male version of the 57-item Portrait Values Questionnaire  to 4 . The behavior items match their corresponding values. For example, “Keep promises I made to friends or family” matches benevolence-dependability value; “Learn something simply for the joy of learning” matches self-direction-thought value; “Look for exciting activities to break up my routine” matches stimulation value.We adopted the English version of Everyday Behavior Questionnaire ” to “do things that provide sensual pleasure .” Additionally, respondents needed to distinguish between never having even one opportunity to do so (X) and never performing a behavior despite having had at least one opportunity (0). The response “X” was treated as missing data, consisting of <1% in this study. We employed two independent back-translation procedures to translate the EBQ from English to Chinese. Prior to analysis, we replaced missing data with series mean. The internal reliability of the 19 behaviors subscales in this study ranged from 0.585 to 0.793.We used confirmatory factor analysis (CFA) to assess the discriminant validity of the 19 values and behaviors among Southwest minorities. It is impossible to include all 19 latent variables, 57 value items, or 85 behavior items in a CFA model of the whole circle , with a value of at least 0.40. Sixteen of the 19 indexes of quality (IoQs) in Column 6 were >0.70, and three lower values were above 0.60. This demonstrated that the indices of all 19 values were reliable.We checked the correlations among the 19 value factors. Each pair correlated significantly, except for the correlation of power-resources with universalism–tolerance and with benevolence-caring. None of the correlations exceeded 0.80, showing that each of the 19 values could be distinguished by each of the latent factors. Column 5 of We performed the same procedure for the behavior items as in the value items. We allowed a reasonably good fit for at least two of the three criteria, which dropped 22 behavior items because of cross-loadings. Column 4 of Z-score transformations of factors. We based MDS analysis on a custom initial configuration of 19 equal points on the theorized circle to estimate the two-dimensional structures. The centered factor scores were used for the MDS analysis to control the individual differences in using response scales.We used the items retained in the CFA to perform the multidimensional scaling (MDS) analyses to test whether the values of minorities in Southwest China follow the theorized circle. All analyses were performed using SPSS 20.0 MDS Proxscal program , with ordinal proximity transformations, Euclidian distance measures, and As We recaptured the 10 original values based on the observed orders. Conformity, security, power, self-direction, universalism, and benevolence were the six basic values that were split into more narrow values using the refined theory . Eleven of 19 values correlated most strongly with its postulated behaviors than with the other 18 sets of behaviors. For seven values , the correlations with their postulated behaviors were the strongest, followed by the behavior promoted by one of its adjacent values. Security-personal and benevolence-caring value correlated most strongly with its adjacent behavior, followed by its postulated behavior.p < 0.05). Humility and stimulation value, which deviated from the theorized values orders, did not present negative correlations with behaviors motivated by their opposing higher-order values (rs ≤ 0.08).We also checked the correlation of each value with the behaviors that were presumably motivated by the opposing higher-order values in According to Bardi and Schwartz , the colTo validate the moderating role of gender in value-behavior relations, we used the mean factor scores of values and behaviors, standardized gender, and formed the interaction terms between values and gender. We regressed each of the 19 behaviors on its corresponding value, gender, and the interaction term.Seventeen of 19 values predicted their corresponding behavior significantly, as shown in Column 1 of Each of the 19 values of minorities in Southwest China formed a single factor on the circular motivational continuum, which provided cross-cultural evidence on predictive and discriminant validity of the theory of refined value. However, our observed orders did not exactly correspond with the theorized orders  means to be a man (ren)The “Five Constants” are the core virtues in the Chinese traditional value system, among which “benevolence” ranks first. It emphasizes that an individual should be humane to others, that is, “Humility value appears inside an unexpected polar angle, together with self-direction-action value. The result accorded with some reports that humility is undergoing a transformation from traditional self-insignificance to that with a modern sense of not renouncing self-interest based on mutual and equal benefit  ear,” a unique dialect in China Sichuan, is used to describe a man who is suggestible, or more obedient to his wife, or “henpecked.” This shows that women are more involved in decision-making and executive than men in the family, which implies their stronger relation of self-direction value to behaviors. Our results may be just by chance but they embody the regional cultural characteristic in Southwest China.Gender difference in power and benevolence domains did not appear in our study. Li and Chen  suggesteThere are some limitations that need to be noted. First, we only examined self-reported retrospective behaviors. Such reports probably attenuated some of the presented value-behavior associations due to incomplete memory and such motivated biases as consistency seeking or social desirability. However, self-reporters can base themselves on the full range of their experiences but not the shared experiences with the target or based on hearsay. Specific behavior frequency of self-ratings during the past year has also been shown to be quite accurate indicators (Gosling et al., With the aim of exploring psychological features of Chinese minorities against diverse cultural context, we examined the relations of values to behaviors among minorities in Southwest China by confirming the predictive and discriminant validity of the refined value theory. Despite some limitations, this study is a pioneering step in understanding the relationship between the values and behavior of Chinese minorities. The strongest relations of 11 of 19 values to their postulated behaviors and the consistent value-ordering with the theorized circle provide cross-cultural evidence to the universality of the refined value theory; however, such seemingly contradictory values as conformity, humility, self-direction, stimulation, benevolence, and tradition motivated more harmonious behaviors in Chinese minorities, in which perhaps the “Golden Mean” asserted by Confucianism rather than the normative pressure plays a role. Our results did not explain the moderating role of normative pressure on value-behavior relations. The moderating role of gender in the value-behavior relations also reflected the regional cultural uniqueness of minorities in Southwest China. The result from this study contribute to explaining ethnic behaviors corresponding to value, and provide psychological evidence to understand the uniqueness of Southwest minorities.There are 56 ethnicities in China, including 55 officially defined ethnic minority groups and one majority group, ethnic Han. These ethnic minorities are so named because the population of each minority is much smaller than that of the Han ethnicity, which is the most populous ethnic group of China and represents the Chinese universal and mainstream culture. For instance, according to the Chinese sixth population census data (Chinese National Bureau of Statistics, The study was carried out in accordance with the recommendations of the Ethics Committee of Southwest University with written informed consent from all participants before the surveys. All participants gave written informed consent in accordance with the Declaration of Helsinki. The protocol was approved by the Ethics Committee of Southwest University.YY and YZ conceived and designed the study. YY collected and analyzed the data, and drafted the initial manuscript. JZ contributed to the data analyses. PF reviewed the manuscript. GJ and YL participated in issuing and collecting the questionnaires. YZ provided the critical comments and revision. All authors approved the final version of the manuscript for submission.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The aim of this study was to investigate the role of kinase-insert domain-containing receptor (KDR) in intrauterine adhesions (IUA) and its mechanism. The Case group consisted of 92 patients diagnosed with IUA, and the Control group included 86 patients with uterine septum who had normal endometrium verified with an uteroscope. In addition, 50 rats were randomly assigned into Control, Sham, Model, NC-siRNA, and KDR-siRNA groups. Rats in the Model, NC-siRNA, and KDR-siRNA groups were induced by uterine curettage and lipopolysaccharide (LPS) treatment to establish the IUA model. Then, immunohistochemistry was applied for detection of VEGF and KDR expression, HE staining was used for observation of the endometrial morphology and gland counting, Masson staining for measurement of the degree of endometrial fibrosis, and qRT-PCR and western blot for the expression of KDR, VEGF, MMP-9, as well as TGF-β1/Smads pathway-related proteins. Compared with the Control group, the mRNA and protein expressions of KDR were significantly higher in IUA endometrial tissues, and the expression of KDR was positively correlated to the severity of IUA. In addition, the injection of si-KDR increased the number of endometrial glands, reduced the area of fibrosis, inhibited mRNA and protein expression of KDR and VEGF, up-regulated the expression of MMP-9 and Smad7, and decreased the expression level of TGF-β1, p-Smad2, p-Smad3, and Smad4 in rats with IUA. Highly-expressed KDR was related to patients' severity of IUA, and silencing KDR may prevent the occurrence and development of IUA via TGF-β1/Smads signaling pathway and up-regulating the expression of MMP-9. Intrauterine adhesions (IUA), also identified as Asherman's syndrome, are caused by the abnormal healing of the injured endometrial basal layer owing to a consequence of trauma or infection to the endometrium, and the clinical symptoms of IUA include menstrual disturbance (amenorrhea/hypomenorrhea), cyclical abdominal pain, infertility, and recurrent pregnancy loss ,2. AlthoVascular endothelial growth factor receptors (VEGFRs), a series of transmembrane receptor tyrosine kinases, could bind VEGF to initiate a signaling cascade that culminates in cellular migration, mitosis, and proliferation, thereby promoting angiogenesis and inducing fibrosis of endometrium . To dateTherefore, we detected the KDR expression in endometrial tissues with or without IUA, and further analyzed the expression of TGF-β1/Smads signaling pathway by the injection of si-KDR into the uterus after establishing the IUA rat model to explore the mechanism of KDR in IUA formation in depth.The experiment in this study was approved by the Ethics Committee of our hospital and performed in obedience to the Helsinki declaration . All speFrom May 2016 to May 2017, 92 patients diagnosed and confirmed with IUA in our hospital were included as the Case group . Based on the American Fertility Society classification of IUAs . The blocking solution of normal goat serum was added at room temperature for 20 min, followed by 1 h incubation with KDR  antibody at 37°C, washing with PBS, and another 1 h of incubation with the secondary antibody at room temperature. After that, tissue sections were washed again with PBS and developed with diaminobenzidine (DAB) to observe the coloration under a microscope. Next, hematoxylin was used for 2 min of re-dyeing, followed by routine dehydration, hyalinization, mounting, and microscopic examination. The statistical analysis of positive rate was conducted based on the classification by Fromowitz et al. : 0 pointEscherichiacoli 055: B5; Sigma, USA) was inserted through the uterine cavity after the curettage using a round surgical needle. Then, the plasmids  were immediately injected into five different sites of the uterus  injury using uterine curettage and lipopolysaccharide (LPS) surgical suture to establish the model of IUAs , showing highly fibrous adhesions according to the record of Li et al. . Next, fThe endometrial tissues of rats were fixed for 24 h in Davidson's solution before the routine procedures of dehydration, hyalinization, wax dipping, and paraffin embedding. Next, the ultra-thin semiautomatic microtome  was used to slice ten 3-μm-thick serial sections, which were baked at 50°C for 1 h, stained with hematoxylin and eosin, and observed under a microscope  to evaluate the pathological changes of endometrial tissues. Then, conventional Masson's trichrome staining  was performed on tissues. After that, five visual fields were chosen randomly from the stained tissues under the high-power lens, and the software Image Pro Plus 6.0  was used for the analysis of pictures of pathological tissues. The number of endometrial glands and degree of endometrial fibrosis were microscopically evaluated as described previously .TM RT reagent Kit . The reverse transcription system was 10 μL in volume, and the conditions for reaction were as follows: 16°C for 30 min, 42°C for 30 min, and 85°C for 10 min. The SYBR® premix Ex Taq TMPCR kit  was used for qRT-PCR detection and the reaction conditions were: pre-denaturation for 2 min at 95°C, and 40 cycles of denaturation for 5 s at 95°C, annealing for 5 s at 60°C, and extending for 30 s at 72°C. The expression of mRNA was calculated using 2-△△Ct with GAPDH as the internal reference gene.The endometrial tissues were ground with normal saline, and the RNA extraction kit  was used to extract the total RNA of each tissue sample. RNA purity and concentration were determined by the UV spectrophotometer  and observed for integrity by agarose gel electrophoresis. The primers of VEGF and KDR were designed using the software Primer 5.0  and synthesized by Sangon Biotech Co., Ltd. (China). cDNA was generated through the reverse transcription of the total RNA using Primescriptg for 15 min at 4oC. Next, the upper supernatant was collected for SDS-PAGE electrophoresis. The proteins were electrically transferred to the nitrocellulose membrane for blocking with 5% skim milk-PBS solution at room temperature. One hour later, the primary antibodies were added for overnight incubation at 4°C, including VEGF , KDR , TGF-β1 , pSmad3 , Smad3 , pSmad2 , Smad2 , Smad4 , Smad7 , MMP-9, and β-actin  . The secondary antibody crosslinked with HRP was added at room temperature for 1 h after the membrane was rinsed with PBS buffer three times. Blots were then developed by enhanced chemiluminescence (ECL) detection. With β-actin as the loading control, the gray value ratio of target band to reference band was regarded as the relative expression level of proteins.The endometrial tissues of each group were added with lysate, homogenized, and centrifuged at 1,200 All data were analyzed using the statistical software SPSS 20.0 . The measurement data are reported as means±SD, with the difference between two groups compared by least significant difference (LSD) test, or among multiple groups tested by one-way ANOVA. Enumeration data were analyzed and tested by chi-squared test or Fisher's exact test, and the relationship between variables was assessed by Spearman's correlation analysis. A P value less than 0.05 was regarded as statistically significant.The KDR protein identified by immunohistochemical staining  was mainThe endometrial cavity surface of the Control and Sham groups was covered by columnar epithelium with abundant endometrial glands arranged in round or oval shape observed by HE staining. Masson staining showed a few stromal collagen fibers and wavily arrayed. Compared with the Control and Sham groups, the rats in the other groups showed different degrees of IUA, a disappearance of endometrial epithelium, and a reduction of glands with tissue hyperplasia, as well as an increase of collagen fibers . CompareCompared with the Control and Sham groups, the expressions of both KDR and VEGF at the mRNA and protein levels were enhanced in the endometrial tissues of rats in the Model group . However, those rats in the KDR-siRNA group presented down-regulated KDR and up-regulated VEGF. In addition, the expressions of KDR and VEGF were significantly lower in the KDR-siRNA group than in the Model group and NC-siRNA group (both P<0.05, As displayed by western blot in Currently, the clinically therapy for IUA is effective for the restoration the uterine shape, but the restoration of the physiological function of the uterus is not ideal . Thus, iTo further investigate the mechanism of KDR in affecting IUA, we constructed the IUA rat model using phenol mucilage and injected KDR-siRNA or NC-siRNA into the uterus of rats. At the 9th day after model establishment, we found increased levels of KDR and VEGF, as well as an enlarged fibrosis area in the rats from the NC-siRNA group, which further confirmed the clinical result and the hypotheses mentioned previously. On the contrary, the elevation of gland number and the reduction of fibrosis area in the endometrium were discovered in the IUA rats treated with si-KDR. Interestingly, there was evidence of a close relationship between VEGF-induced cell proliferation and adhesions and the KDR downstream signaling pathways , and ourOn the other hand, among various fibrosis-related cytokines, TGF-β, a pleiotropic cytokine, was well-accepted as the promoter of fibrotic lesions , which cCollectively, increased KDR was found in the endometrium of IUA patients, which was positively related to IUA severity. However, after silencing KDR, the expression of KDR and VEGF declined and the TGF-β1/Smads pathway was greatly inhibited, with the elevation of MMP-9 and the reduction of fibrosis, thus decreasing the occurrence and development of IUA.


Triarrhena sacchariflora Nakai has been considered a promising energy crop for utilization in ethanol production. Flooding stress is among the most severe abiotic stressors in the production of Nakai. However, the physiological and molecular biological mechanisms of Nakai response to flooding is still unclear. In the present study, in order to understand the molecular mechanisms of Nakai in response to flooding stress, the transcriptome, physiological and biochemical were investigated.In recent decades, the frequency of flooding is increasing with the change of global climate. Flooding has become one of the major abiotic stresses that seriously affect growth and development of plants. The results demonstrated that significant physiological changes were observed in photosynthetic system, antioxidative enzyme activity, chlorophyll, carotenoid, proline, lipid peroxidation and soluble sugar content under normal and flooding treatments. Such as, the chlorophyll, carotenoid contents and photosynthetic system were significantly decreased. Whereas, the antioxidative enzyme activity, proline, lipid peroxidation and soluble sugar has increased first and then decreased under treatments compared with the normal plants. Additionally, a total of 8832, 6608 and 3649 unigenes were validated to be differentially expressed under different treatments, respectively. Besides, gene ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis of the different expression levels of genes also presented processes, which involved in photosynthesis, sucrose catabolism, glycolysis, stress response and defense, phytohormone biosynthesis and signal transduction.The results provide a comprehensive view of the complex molecular events involved in the response to flooding stress of Nakai leaves, which also will promote the research in the development of flood-resistant crops and provide new tools for Nakai breeders. Triarrhena sacchariflora (Maxim.) Nakai is a large perennial herbaceous plant, belonging to the Miscanthus spp. of Gramineae, which is widespread in the northern and midland of China [of China , 2. Nakaof China , 3. Meanof China , 5. Therof China , 6.), are changed under flooding stress due to the disturbed balance of ROS generation and removal [With the change of global climate, the frequency of flooding is increasing. Flooding has become one of the major abiotic stresses that seriously affect growth and development of plants , 7. The  removal –23. High removal , 25.Arabidopsis thaliana [Arabidopsis thaliana [In recent decades, the physiological and molecular mechanisms of adaptation to flooding stress in many plants have been investigated , 11, 13.thaliana . Howeverthaliana , 33–36. thaliana , rice [3thaliana , soybeanthaliana , 40, maithaliana , canola thaliana  and so oTriarrhena sacchariflora (Maxim.) Nakai leaves. Moreover, we examined the accumulation and change of photosynthetic rates, photosynthetic pigments, proline, antioxidative enzyme activity, lipid peroxidation (MDA) and soluble sugars. The results described the changes of physiological, biochemical and the complex molecular mechanisms in response to flooding stress at different treatment times, which is helpful to our understanding of plants response to flooding stress.In the present study, we analyzed the physiological and transcriptome responses to different levels of flood treatment in Meanwhile, these transcriptome databases also will promote the research in the development of flood-resistant crops and provide new tools for Nakai breeders.To dissect the influence of Chlorophyll and carotenoid contents under different times of flooding stress, we determined the chlorophyll a, b, total and carotenoid content of normal (0-DAF) and after samples treatment for 2, 4, 6 and 8 days after flooding (DAF). The results showed that the chlorophyll a, b, total and carotenoid content were significantly decreased with the continuation of flooding treatment Table . In addiLeaf gas exchange in Naka was measured for 0, 2, 4, 6 and 8 DAF, starting at the onset of flooding treatment , superoxide dismutase (SOD) and total peroxidase (POD) were evaluated Fig. a-c. Acco. Lipid peroxidation levels increased to the maximum at 2-DAF. The content of Proline also changed similarly to that of MDA. They increased at first and then decreased significantly when the periodical flooding time was longer than 6 days . Therefore, to further exploration the physiological responses and the transcriptome information profiles of the three treatments in Nakai, we performed a set of consecutive and comprehensive transcriptome sequencing from the 3 treatments samples of flooding stress. A total of 102,590,796, 111,241,228 and 108,426,642 clean reads were obtained from the L0- DAF, L2-DAF and L4-DAF samples, respectively  between the normal and treatment samples, all assembled unigenes were analyzed using the edgeR software, with an FDR < 0.05 and |logFC| ≥ 1 as a cutoff. A total of 8766 (0_2-DAF: 0-DAF vs 2-DAF) and 6608 (0_4DAF: 0-DAF vs 4-DAF) genes were identified as being differentially expressed at normal samples, relative to treatment samples, respectively Fig. , which cIn order to classify the functions of the DEGs of Nakai, these DEGs were assigned based on Gene ontology (GO) analysis. The results were shown in Additional file To further evaluate these DEGs of Nakai, we considered the biological function of the DEGs that were regulated in response to flooding. These DEGs of 0_2-DAF, 0_4-DAF and 2_4-DAF were performed to enrichment analysis based on GO and KEGG pathways. GO enrichment analysis of DEGs indicated that 324, 182 and 143 GO terms were significantly enriched with the criteria of FDR < 0.05, respectively , 34, 35 and 30 KEGG pathways were significantly enriched between the normal and treatment samples, respectively , 21jasmonic acid (JA),18 abscisic acid (ABA), 9 cytokinine (CK), 7 ethylene, 7salicylic acid (SA), 7 brassinosteroid (BR), 4 gibberellin (GA) genes  homology search against the PlantTFDB. These TFs were classified into 32 TF families in this dataset . The Chlorophyll-chlorophyllido hydrolase 2 gene was significantly up-regulated at 4 days of flooding . A total 50 DEGs (down-regulated) were detected from the photosynthesis (Ko00195) and photosynthesis - antenna proteins (Ko00196) pathway, which is consistent with a reduction of assimilation rate. Stomata play a pivotal role in photosynthesis by maintaining the balance of gas exchange between the aerial parts of the plant and the atmosphere [On the other hand, when plants are subjected to flooding stress, both light and carbon supplies are limited due to the slower diffusion rates in water, which also reduces plant photosynthesis , 46. Hermosphere , 48. Witmosphere , 49, 50.Z. mays [C. indicum [S. indicum [T. aestivum [2 and H2O2 using disproportionation reaction induced by SOD. CAT and POD can reduce or eliminate the excess H2O2 to prevent peroxidation, which can enhance the ability of plants to adapt to flooding stress [Flooding condition lead to hypoxic and anoxic conditions for plants and triggers the accumulation of ROS, which is many causing severe damage to plant cells , 44, 47.Z. mays , C. indi indicum , S. indi indicum  and T. aaestivum . Superoxg stress . MeanwhiHowever, these antioxidant systems cannot maintain the balance of reactive oxygen systems in plants for a long time. When the oxygen activity system is out of balance, lipid peroxidation is an indicator of oxidative damage under flooding stress. Malondialdehyde (MDA) is a major product of lipid peroxidation, and its increase reflects damage to plant under flooding . The resRumex palustris, which indicated the induction of ethylene can be inhibited under flooding. Besides, ABA is also involved in the process of plant flooding stress [Plant hormones play a crucial role in the process of plant growth and development, as well as in the response to biotic and abiotic stresses , 60, 61.g stress . It is pg stress . In thisArabidopsis thaliana. On the other hand, the specific mechanism of CK action under flooding stress in plants are not fully known. But Zalewski et al. (2010) reported that the productivity of barley is increased by silencing the CK oxidase gene under normal as well as stressed conditions [In addition to ethylene and ABA associated DEGs response to flooding stress, DEGs associated with the IAA and CK metabolism were also observed. IAA metabolism was observed with 9 DEGs identified during flooding stress, including 6 Aldehyde dehydrogenase, two indole-3-pyruvate monooxygenase (YUCCA) and a BUD13 homolog genes. Present studies suggest that IAA, through ARF gene family, activates stress specific auxin response genes in order to mitigate the negative effects of abiotic stresses , 66. Auxnditions . These rnditions , 68.Transcription factors play an important regulator of the response to various biotic or abiotic stresses in plants. Such as drought, flooding, heat or cold and salinity. Some studies reported that these TFs are involved in various abiotic stress and positively improve plants tolerance –72. Thusstarch and sucrose catabolism, glycolysis and ethanoic fermentation to increase substrate-level production of ATP [Cajanus cajan genotypes was been reported under waterlogging condition [Dynamic changes of metabolic pathways in response to submergence, flooding and waterlogging have been reported in a wide range of plants. In general, with the photosynthesis limiting, anaerobic respiration starts to become the primary pathway for supplying energy to the plants. Such as, n of ATP , 11, 13.n of ATP . Such asondition . So, ourondition , 76. Othondition .The lack of information available about the molecular mechanisms involved in flooding stress responses in Nakai is currently a major constraint for the further development. In this study, we provided the first characterization of the physiological responses and the transcript profiles of under different levels of flooding treatments in Nakai leaves. Moreover, we examined the accumulation and change of photosynthetic rates, photosynthetic pigments, proline, antioxidative enzyme activity, lipid peroxidation. Besides, many of the unigenes were identified in the present study, which have the potential to be used for the development of novel Nakai varieties with improved productivity and stress tolerance. Thus, the results provide a comprehensive view of the complex molecular events involved in the response to flooding stress of Nakai leaves, which also will promote the research in the development of flood-resistant crops and provide new tools for Nakai breeders.Triarrhena sacchariflora (Maxim.) Nakai was cultivated in the greenhouse of Wuhan University . Plants were cultured with rhizome in pots, each of which contains at least four rhizome of about 10 cm in length. A total 150 potted Nakai plants were cultured in a greenhouse under 400-watt high intensity lamps with a 16 h day and 8 h night photoperiod at a temperature of 23° ± 3 °C. Two months later, we selected 100 potted plants of similar size and divided them equally among two groups. After that two groups of plants were transplanted into same tanks, respectively. The plants in each tank were design consisting of different treatments  and fore harvest time points: 2-, 4-, 6- and 8-days after flooding (DAF). Leaf from the normal were used as 0-DAF sample and those from the flooding conditions were considered as 2-DAF, 4-DAF, 6-DAF and 8-DAF sample. Each sample was the mixture of leaf from 3 plants and three biological repeats were taken from each period. After each sampling, the sample tissue was immediately frozen using liquid nitrogen and stored at 80 degrees Celsius before use to RNA extraction and other treatments.In order to understand the difference in photosynthetic indexes between normal and flooding condition plants, the photosynthetic efficiency-related parameters were made by a portable photosynthetic system CIRAS-3. Actinic light was supplied by light-emitting diodes . All measurements were carried out between 09:00 and 10:00 h and carried out six biological duplication. Besides, chlorophyll and carotenoid contents were determined as the method from Wu and Yang .The samples (0.2 g) were used for enzyme extraction. Firstly, we grind the sample with 5 ml sodium phosphate buffer (pH 7.8). Secondly, the sample after grinding was centrifuged at 12,000 g for 20 min. After that, we collect the supernatant will used for determination of enzyme activity. For CAT activity was performed as described previously –45. For Total RNA of six sample were acquired using HiPure Plant RNA Kits . We verified and purified the RNA quality by NanoDrop ND-2000 spectrophotometer  and Dynabeads® Oligo (dT) 25 , respectively. After that, a cDNA library was generated using the purified mRNA, which was used for cluster generation using the TruseqTM RNA sample prep Kit and were subsequently sequenced using the HiSeq 4000 SBS Kit.To obtain clean reads, the SeqPrep and Sickle software were used to filter the raw data. The clean reads were assembled using Trinity software to generate the unigenes. To identify potential protein-coding genes and annotate the function of these unigenes, all assembled sequences were screened for ORF prediction using EMBOSS software. The predicted protein-coding sequences were searched in the Nr (NCBI non-redundant) database, Pfam database, KEGG (Kyoto Encyclopedia of Genes and Genomes pathway) database, String database, GO (gene ontology) database and Swiss-Prot database using BlastX . The proteins with the highest matching value were considered to contain annotation information.P-Value < 0.05.In order to test whether the variation of biological duplication between samples accords with the expectation of experimental design, sample correlation and PCA analysis were adopted. Principal component analysis (PCA) and hierarchical clustering were performed to assess transcriptome similarity among samples and to evaluate sampling between biological replicates. To analyze the differential expression of gene/transcript between different samples, the RSEM software was used to calculate the expression of each unigenes. Here, we use the RPKM value to analysis the differential expression between different samples by the edgeR software, filter parameters are as follows: FDR < 0.05 and |log2FC| ≥ 1. Besides, GO enrichment and KEGG enrichment analysis was performed on the differentially expressed unigenes using Goatools and KOBAS software, with Corrected Transcription factors are proteins that bind specifically to DNA sequences and typically has one or more domains that bind to DNA. Transcription factors not only bind to promoter regions in the upstream of genes, but also constitute transcription factor complexes with other transcription factors to affect gene transcription, and thus play an important role in organisms. To get cognate transcription factor information of Nakai, all unigenes were examined from the plant TFDB database by the HMMER analysis method.In order to verify the results of transcriptome datas, we selected eight unigenes with significantly different expressions for the expression level verification in the samples by qRT-PCR. We use the primer 6.0 software to design the primers  was used to test the flooding effects on the ecophysiological performance of Nakai. Significant differences between control and treatment were determined for physiological and biochemical using SPSS 19.0 .One-way analysis of variance (ANOVA) with the least significant differences  functional analysis of DEGs between normal and treatment samples.Additional file 3: Figure S3. Heatmap analysis of DEGs related to plant hormone signal transduction. The expressions of gene are displayed different colors. Red means high expression, and blue means low expression.Additional file 4: Figure S4. Heatmap analysis of DEGs related to 205 TFs in different samples. The expressions of gene are displayed different colors. Red means high expression, and blue means low expression.Additional file 5: Figure S5. Heatmap analysis of DEGs related to photosynthesis and photosynthesis - antenna proteins pathway. The expressions of gene are displayed different colors. Red means high expression, and blue means low expression.Additional file 6: Figure S6. Heatmap analysis of DEGs related to peroxisome pathway. The expressions of gene are displayed different colors. Red means high expression, and blue means low expression.Additional file 7: Table S1. The summary of tag numbersAdditional file 8: Table S2. The unigenes annotation in different public protein database Additional file 9: Table S3. The unigenes expression statistics Additional file 10: Table S4. The differentially expressed genes (DEGs) in response to floodingAdditional file 11: Table S5. Gene ontology (GO) functional enrichment analysis of DEGs between normal and treatment samples.Additional file 12: Table S6. KEGG pathways enrichment analysis of DEGs between normal and treatment samples.Additional file 13: Table S7. Transcription factors response to flooding Additional file 14: Table S8. Primer sequences used in the qRP-PCR experiment


To tackle the problem of obesity and related diseases in Switzerland, cost-efficient, effective, and innovative primary health care interventions for weight management are required. In this context, Oviva has developed a scalable technology for registered dietitians to counsel overweight and obese patients via a mobile phone app.The aim of this study was to evaluate the effectiveness and feasibility of weight loss counseling by dietitians using a mobile phone app for patients with overweight and obesity.1c, fasting glucose, fasting insulin, triglyceride, high-density lipoprotein cholesterol, blood pressure (BP), body mass index (BMI), waist circumference, body fat, and responses to a self-administered questionnaire with questions regarding participants’ physical activity, dietary assessment, and health-related quality of life. Changes were tested at baseline, after 3 months, and after 12 months, as well as between the third and the 12th month.In this pre- and posttest pilot study, overweight and obese adults participated in a 1-year behavioral intervention to lose weight through remote counseling by dietitians in the German-speaking part of Switzerland. The study started in April 2016 and finished in May 2018. Participants received individual counseling through the app and the exchange with the dietitian focused on regular feedback on photo-based food log, motivation, and education. The contents were tailored to the individual lifestyle goal set. The predefined intensity of remote counseling decreased during the year. Group chat could be used. The outcomes examined were changes in weight (primary outcome), hemoglobin AP<.001), between week 12 and week 52 it was −1.1 kg (range: −9.7 to 7 and P=.08), and the median change during the entire period of intervention was −4.9 kg (range: −21.9 to 7.5 and P<.001). Furthermore, changes in BMI, waist circumference, body fat, and BP between baseline and 12 weeks and between baseline and 52 weeks were also significant. Significant changes in certain eating habits were also demonstrated .In total, 36 women and 7 men, with a mean age of 40.6 years, participated and 36 participants completed the study. Median weight change after the first 12 weeks was −3.8 kg  Obesity has become one of the biggest public health challenges in recent decades . In EuroInternational guidelines recommend a weight loss of at least 5% for the treatment of individuals with overweight or obesity to achieve positive clinical effects. The magnitude of weight loss is associated with dose-effect improvements in cardiovascular risk factors such as high blood pressure (BP), hyperlipidemia, and hyperglycemia . In addiTherefore long-term, cost-efficient, effective, and innovative primary health care interventions for weight management are needed.New digital technologies could be a possible solution as they provide innovative counseling possibilities with greater flexibility to coach overweight and obese clients. Such technologies can help reach clients who are unable and/or unwilling to engage in face-to-face counseling and make it possible to assist clients more conveniently and as required . Studies show that both face-to-face and remote counseling are effective methods for losing weight, as well as for maintaining weight loss -20. MobiAlthough these innovative tools offer dietitians new potential for counseling and supporting eating and physical behavior change, their use of these tools remains limited. One reason may be that few studies on the long-term impact of Web-based nutrition counseling exist. The number of commercial health and nutrition apps available is huge; however, specific apps for remote counseling, which are aligned to the needs of a dietitian, are limited. In Switzerland, Oviva, a new type of health care provider, has come to the market using new technology to scale the dietitian workforce. Oviva has developed a scalable technology for dietitians to counsel overweight and obese patients via a mobile phone app. Through this app, dietitians can organize their sessions through messaging and video calls, have access to photo food diary, activity, and weight tracking features allowing them to monitor client progress through their photos, activity, and weight logs, as well as set goals and share documents and surveys. Assessing the implementation of this novel kind of intervention is important ,26 as siThis study was carried out as a 1-year, single arm pre- and postpilot intervention with overweight or obese adults in Switzerland from March 2016 (first participant in) to May 2018 (last participant out). All study procedures were approved by the local institutional review board and the study is registered under ClinicalTrials.gov (NCT02694614). Follow-up will be conducted 1 year after the intervention.Sample size was calculated with a significance level of 0.05 and power of 80%. A sample size of 36 was needed to detect a weight loss of 0.5 SD, which corresponds to a medium effect size according to Cohen, using a 2-sided Wilcoxon signed rank test. To account for dropouts, 50 participants were planned. After screening, 43 participants were included in the study.2, fluent in German, mobile phone user (iOS or Android), and capable of sending and receiving SMS text messages and pictures. Participants were excluded if they were pregnant or breastfeeding, were diagnosed with conditions other than dyslipidemia, hypertension, and insulin resistance requiring nutrition therapy, had serious disease requiring continuous drug therapy, were on a weight reduction diet during the last 6 months, took medication for weight loss in the past, or enrolled in another weight loss program. Subjects were invited to participate with flyers distributed through the Center for Obesity and Metabolism Medicine Winterthur (in Canton Zurich), via general practitioners, advertisements on the websites of the participating research institutions, local newspapers, and through word of mouth advertising. Interested persons received written and verbal information about the study. Screening took place following informed consent to determine the eligibility of participants on the basis of the inclusion and exclusion criteria at the Center for Obesity and Metabolism Medicine Winterthur.Participants were included in the study if they met the following inclusion criteria: adults (aged 18 years and over), body mass index (BMI) between 26 and 33 kg/mThe program is designed to facilitate personal coaching by registered dietitians. This digital communication system connects patients remotely with their dietitian who has all the tools required for remote counseling. Via customer-facing apps for iOS and Android, all clients are able to collect relevant information  and communicate easily with their dietitian. All features of the app are modeled on typical activities in a dietitian’s everyday practice and include chat-like communication with dietitians, group chats for support from peers, dietitian’s profile to create a more personal connection, a photo-based food log, activity and weight logs, a goal scorecard, showing past goals and future options, a content database, feedback to the dietitian, and links to standard learning materials. Client exchanges with the dietitian focused on regular feedback regarding the photo-based food log, motivation, and education . Clients were also able to join group sessions within the same app. All data were transferred and saved via secure channels. For the dietitians, all information was displayed in a secure cloud-based platform, similar to Webmail. They could review all relevant information for each client individually, take notes, create customized surveys, and use a content database, including relevant links. This behavior-based intervention was carried out entirely remotely by 3 registered dietitians using Oviva technology. The intervention comprised 3 different coaching phases as described in The content of the remote counseling was tailored to the specific needs of each participant, but it was tailored with the overall aim to lose weight. The behavioral strategies used were self-monitoring, goal setting, stimulus control, elaboration of alternative strategies, social support, positive reinforcement, and relapse prevention (particularly toward the end of the intervention). Motivational interview techniques were used throughout the entire intervention process.Key counseling topics were based on the needs of the participants and included negative energy balance, plate model , portion size, meal rhythm, energy density of food, quality of carbohydrate, fat, carbohydrate-modified diet, adequate protein sources, dealing with special situations , grocery shopping, and physical activity.1c (HbA1c), fasting glucose, fasting insulin, triglyceride, high-density lipoprotein (HDL) cholesterol, and BP. Measurements were carried out by a clinical nurse in the morning. Participants were instructed to fast overnight (8 hours), abstain from smoking, consuming alcohol, and drinking caffeine-containing beverages, as well as from vigorous exercise 48 hours before blood samples were taken and body composition was measured. Weight was measured using a clinically validated and calibrated scale . Participants were lightly dressed, without shoes, and weight was recorded to within 0.01 kg. Height was measured using a calibrated stadiometer (Forma Seca). The measurement was carried out in an upright position with feet positioned on the floor board of the stadiometer. Heels, buttocks, and the back of the head touched the back board of the stadiometer with arms on the sides. BMI was calculated by dividing body weight in kilograms by height in meters squared. For the measurement of the waist circumference, an inelastic tape was placed directly on the skin between the lower rib and iliac crest. Each measurement was performed twice after exhalation and recorded within 0.5 cm. If the difference between the 2 measurements was greater than 1 cm, a third measurement was performed and the mean of the 2 closest measurements was calculated. Body composition was measured by bioelectrical impedance analysis  with participants in upright position, lightly dressed, and barefoot. BP measurements were performed using the Riva-Rocci method with a special instrument (Firma boso-medicus SN 768 00 440 729 Bosch+Sohn GmbH und Co) and a stethoscope. The measurement was performed 3 times using the right arm after participants had been sitting for at least 10 min. The average of these measurements was used for analysis. Blood samples were obtained after overnight fasting from the antecubital vein to determine HbA1c, fasting glucose, fasting insulin, triglyceride, and HDL cholesterol. All laboratory assays were performed by Labor Risch Schaffhausen using standardized methods as described in the study protocol.The primary outcome was defined as weight change between baseline (M0) and 1 year (M12). All measurements were examined at baseline (M0), at 3 months (M3), and 1 year (M12), and it included anthropometric, clinical, and questionnaire assessments. Anthropometric and clinical assessments included body height, body weight, BMI, waist circumference, body fat, hemoglobin APrephase or initiation (week −2 to 0)Complete initial assessment; implementation: photo-assisted dietary and activity recording; setting overall aim.Phase 1: Transition (month 1 to 3)Mobile phone assisted patient coaching 5 times per week; feedback on nutrition and activity; setting 1 or 2 specific goals for 2 weeks at the time; assessing goals and adapting them if necessary; providing information and education materials that are appropriate to the goals; 1 Skype call at the end; clinical assessment at the end.Phase 2: Stabilization (month 4 to 6)Mobile phone assisted patient coaching 3 times per week; feedback on nutrition and activity; strengthening new behavior; setting new goals if necessary; assessing goals and adapting them if necessary; providing information and education materials that are appropriate to the goals; exchange with peer groups .Phase 3: Maintenance (month 7 to 12)Mobile phone assisted patient coaching once every 2 weeks; feedback on nutrition and activity if required; exchange with peer groups ; access to Web-based education materials; final chat; clinical assessment at the end.A questionnaire was used to assess socioeconomic data, dietary assessment, physical activity, and health related quality of life. Physical activity was assessed using the self-administered Global Physical Activity Questionnaire (GPAQ), developed and validated by the World Health Organization. Dietary intake was investigated using a brief 11-item simplified food frequency questionnaire. The English version of this questionnaire was developed and tested in a diverse population as a relatively simple, valid, and efficient tool for dietary assessment . The EngAll outcomes were analyzed on the basis of the intention-to-treat (ITT) population, defined as all participants who started the intervention. Supportive analyses based on the per-protocol (PP) population were performed for selected outcomes. For all outcomes, the changes were calculated from baseline to the specified time points. The changes over the whole observation period were investigated using nonparametric analysis of variance methods for longitudinal data . PosthocFollowing screening, a total of 43 people aged between 20 and 67 years were included in the study. 36 participants completed the 1-year intervention. Overall, the proportion of women was higher  than men, with 6 women and 1 man not completing the intervention. Socioeconomic data of the study population are summarized in 1c level. At the beginning of the study, 3 people took medication to treat hypertension and 2 people took lipid-lowering medication. The medication did not change during intervention.In total, 33 people were initially diagnosed with dyslipidemia, 17 with insulin resistance, and 4 with hypertension. No participant was diagnosed with diabetes and 1 participant was diagnosed with prediabetes because of a slightly elevated HbAP<.001). Weight loss was also significant between M0 and M3  but not between M3 and M12  are shown in 2 [P<.001]) and between M0 and M3 (by 1.4 kg/m2 [P<.001]). Between M3 and M12, the decrease was 0.4 kg/m2 (P=.06). Waist circumference, body fat (kg), or body fat (%) showed similar trends; a significant reduction between M0 and M12 (P<.001), as well as between M0 and M3 (P<.001), but no significant effects between M3 and M12  and blood lipids (triglyceride and HDL cholesterol). In contrast, a significant reduction in systolic BP was observed among all timepoints , as well as in diastolic BP between M0 and M12 . The values for health-related quality of life at month 12 (PCS: 55.4 and MCS: 55.1) showed little change in comparison with the values at month 0 and the changes were also nonsignificant (PCS: P=.58 and MCS: P=.58). The overall result of GPAQ showed a small but nonsignificant increase in total moderate-vigorous physical activity metabolic equivalent minutes per week (MET-min/week) among different assessment times . An analysis according to the 3 subcategories showed the following results. In subcategories activity at work / travel to and from places, no changes were apparent over the entire course of the year, whereas there were significant changes in the category recreational activities during this period . The results for certain eating habits  were significant between M0 and M3 and M0 and M12 and for salty snacks between M0 and M3. The total score for dietary consumption  showed a significant reduction respective improvement toward a healthier diet between M0 and M3 (P<.001) and M0 and M12 (P<.001).The SF-12 score (MCS and PCS) decreased during intervention, especially during the first 3 months (PCS: 53 to 55.2 and MCS: 53.2 to 54.9), but the changes were not significant (PCS: P=.01) [This study investigated whether a 1-year remote counseling intervention by dietitian targeting weight loss was effective and feasible for people with overweight and obesity. The results show that this form of behavior counseling leads to significant weight loss, both in the first 3 months during an intensive remote counseling period, as well as over the entire course of 1 year. Weight loss also occurred between the third and 12th month, although this was not significant. The positive results in this study are in line with other studies investigating Web-based counseling and weight loss ,31-33. IP=.01) ,33. Howe1c, blood glucose, and insulin despite weight loss. One reason for this could be the small sample. The sample size calculation was based on the primary outcome. In addition, for some participants, the initial values of these parameters, particularly HbA1c, were already within a normal range; therefore, a change in these values was not expected. Furthermore, the initially diagnosed cardiovascular risk factors (dyslipidemia or/and insulin resistance) in many participants could also have an impact on these results, although participants were free of serious diseases and only 5 participants took medication to treat hypertension and/or lipid-lowering medication during intervention.Over half of the participants lost at least 5% of their initial weight. This amount of weight loss is typically associated with positive clinical effects, namely an improvement in cardiovascular risk factors. A systematic review demonstrated the following effects of dietary advice: reduction in systolic BP by 2.61 mm Hg (95% CI 1.31-3.91), diastolic BP by 1.45 mm Hg (95% CI 0.68-2.22), and 24‐hour urinary sodium excretion by 40.9 mmol (95% CI 25.3-56.5) after 3 to 36 months, between intervention and control group. However, no changes in HDL cholesterol, triglyceride levels, and fasting glucose were seen [recreational activities subcategory, whereas no changes were observed in the subcategories activity at work / travel to and from places. Increase in recreational activity is important as this form of activity may contribute to the prevention of cardiovascular diseases and cancer [Weight loss and improvement in other parameters were achieved by lifestyle intervention. The most important components of such weight loss interventions include a moderate calorie-reduced diet, increased physical activity, and the application of behavioral strategies to facilitate compliance with nutrition and activity recommendations . In thisd cancer . With red cancer . In thisThe authors attribute the reasons for the successful weight reduction in this study to the following aspects. Dietitians used a wide range of behavioral change and counseling techniques  and applied evidence-based nutritional knowledge according to individual needs . They also provided simple recommendations for physical activity. The results of other studies demonstrate that interventions aimed at changing eating habits or sedentary lifestyle are more effective if they include behavioral change techniques ,40. AnotThe intensity of the intervention typically affects study outcomes . In thisThis study was a 1-arm pilot study; therefore, conclusions concerning effectiveness are limited. Generalization is limited because of the small sample size, individual focus in counseling, and the fact that it was a single-center trial. Randomized control trials are needed to examine efficacy and cost-effectiveness of Oviva and remote counseling in general. Pilot studies are described in the literature as an important first step in assessing the implementation of a novel kind of intervention ,26. As sFor this study, it was important to ensure that the counseling process took place in a real-life setting. For example, the study physician held a brief motivating discussion during the clinical assessment at M3 with the participants about the course of the intervention. This might also have had a positive effect on weight progression. To meet the client's needs, small deviations from the contact frequencies were accepted. However, the tolerable limit was specified in the study protocol. Furthermore, 2 clients requested a change of dietitian as they were dissatisfied. This request was granted to prevent these participants from dropping out of the study. This has been considered in the PP analysis. Nevertheless, this indicates that the working relationship with the dietitian has an important influence on weight loss. In addition to the scientific and practical contents, the working relationship with the dietitian must be developed in a different way in comparison with face-to-face counseling. A qualitative study in which health care professionals were interviewed suggested that it is a challenge to establish and maintain an empathic relationship when conducting electronic health  lifestyle coaching compared with face-to-face coaching . As eHeaA mobile phone app for overweight or obese patients represents a new approach to weight loss counseling and has the potential to make a valuable contribution to tackling this important public health nutrition problem. This study has shown that in addition to the professional skills of dietitians, a profession-specific app such as Oviva can provide effective support that meets the needs of both dietitians and clients on the long path of behavioral change and sustainable weight reduction.


As a result, about 1,363 Gb transcriptome clean data was obtained. After the assembly, we obtain 76,602–154,860 unigenes for each species with sequence N50 length of 1,564–2,143 bp. More than half of the unigenes were functionally annotated by public databases. The Schizothoracinae fishes in this work exhibited diversified ecological distributions, phenotype characters and feeding habits; therefore, the comprehensive transcriptome data of those species provided valuable information for the environmental adaptation and speciation of Schizothoracinae in the Tibetan Plateau.The Schizothoracinae fishes, endemic species in the Tibetan Plateau, are considered as ideal models for highland adaptation and speciation investigation. Despite several transcriptome studies for highland fishes have been reported before, the transcriptome information of Schizothoracinae is still lacking. To obtain comprehensive transcriptome data for Schizothoracinae, the transcriptome of a total of 183 samples from 14 representative Schizothoracinae species, were sequenced and  Machine-accessible metadata file describing the reported data: 10.6084/m9.figshare.11475294 Due to the special geographical environment, many highland species that are distributed in and around the Tibetan Plateau have gradually formed unique characteristics to tolerate harsh living conditions during the long-term evolution2. The Schizothoracinae fishes, members of family Cyprinidae, are endemic to Asian highlands including 15 genera and ca. 100 species3. In China, more than 70 species, account for over 80% of the world’s Schizothoracine fishes, are mainly distributed in lakes and rivers of the Tibetan Plateau and adjacent areas4. According to the morphological characteristics, the Schizothoracine fishes can be divided into three groups: the primitive group, the specialized group and the highly specialized group5. Several researches on the morphology, archaeology and molecular biology of Schizothoracine fishes on the Tibetan Plateau have shown that there is close correlation between the species diversity and the uplift of the Tibetan Plateau7 and the morphological traits of Schizothoracine fishes is related with specific periods of geological evolution of the Tibetan Plateau such as pharyngeal teeth, scales and whiskers5. Therefore, the Schizothoracine fishes are considered as good model species for the investigations on highland adaptation and speciation. More genomic and transcriptome data are required to decipher the relationship of the speciation and the uplift of the Tibetan Plateau for the Schizothoracine fishes.The Tibetan Plateau, the world’s largest and highest plateau, has unique geographical and climatic characteristics, such as the high altitude, dramatic difference in day and night temperature, strong solar radiation8 and Tibetan antelope9). Few study was reported on highland fish, especially for Schizothoracinae fishes. One of the major reasons was the complexity of the genome, such as high content of repeats and polyploidy10. Transcriptome sequencing is a good choice to construct the sequence dataset for transcribed genes in many polyploidy cases11. Despite several transcriptome analyses on highland adaptation have reported in Schizothoracine fishes before16, the species and tissues used for transcriptome sequencing were still limited. There is a great demand for more transcriptome sequencing data for the adaptation and evolution of Schizothoracine fishes in the Tibetan Plateau. In this work, we obtained and released a total of ∼1.36 Tb of high-quality transcriptome data for 183 samples of 14 representative Schizothoracine fish covering 5 genera from 6 drainage systems and 3 lakes in the Tibetan Plateau , Schizothorax lissolabiatus (S. lissolabiatus), Schizothorax nukiangensis (S. nukiangensis), Schizothorax plagiostomus (S. plagiostomus), Schizothorax labiatus (S. labiatus) and Schizothorax davidi (S. davidi). The specialized group contains Ptychobarbus kaznakovi in Ptychobarbus genus. The highly specialized group contains 7 species in 3 genera, such as Gymnocypris namensis (G. namensis), Gymnocypris przewalskii , Gymnocypris eckloni (G. eckloni) and Gymnocypris selincuoensis (G. selincuoensis) of the Gymnocypris genus, Schizopygopsis younghusbandi (S. younghusbandi), and Schizopygopsis pylzovi (S. pylzovi) of the Schizopygopsis genus, Platypharodon extremus (P. extremus) in the Platypharodon genus. The samples were collected from the six major rivers and three lakes of the Tibetan Plateau including Yarlung Zangbo River, Shiquan River, Lancang River, Nujiang River, Jinsha River, Yellow River, Lake Namtso, Lake Qinghai, Lake Siling Co  for a few minutes before the sample collection. A total of 183 tissues were collected from 14 representative Schizothoracine fish in our study, including muscle, liver, spleen, gonads, skin, swim bladder, gut, eye, gill, kidney, heart, brain, blood, fat, vibrissa  according to the manufacturer’s instructions and was determined with a photometer for RNA sample integrity . RNA samples passing the quality criteria  were used for the library preparation and RNA sequencing. All samples were sequenced on an Illumina HiSeq X Ten platform with 150 bp paired-end mode. In preset research, a total of more than 10 billion raw PE reads were obtained from all libraries. After filtering by removal of adaptor sequences, contaminated reads and poor–quality reads, we obtained approximately 1.4 Tb of clean data with Q20 bases larger than 96.94%. The average of 7.6 Gb sequencing data were obtained for samples , including NCBI nonredundant protein database (NR), Swiss-Prot, KEGG pathway database and KOG database. Only the best hits with the highest sequence homology was used for annotation. Then, the gene ontology (GO) terms analysis of the predicted protein based on the NR in NCBI was performed with the Blast2GO software version3.1 with default parameters. We found that at least 40.2% of unigenes per species were annotated based on proteins in four public databases . The total amount of RNA, RNA integrity and rRNA ratio were used to estimate the quality, content and degradation level of RNA samples. In the present study, RNAs samples with a total RNA amount ≥ 10 μg, RNA integrity number ≥ 8, and rRNA ratio ≥ 1.5 were finally subjected to construct the sequencing library.38. Firstly, adaptors in the reads were filtered out; secondly, reads with more than 10% of N bases were filtered out; thirdly, reads with more than 50% of the low-quality bases  were filtered out. If any end of the pair was classified as low quality, both pairs were discarded. The initially generated raw sequencing reads were also evaluated regarding quality distribution, GC content distribution, base composition, average quality score at each position and other metrics.The raw sequencing reads generated from the Illumina platform were rigorously cleaned by the following procedures as in the previous studySupplementary Table S1Supplementary Figure S1