Objective:To develop a method of employing flow cytometry to directly detect the γ-H2AX expression levels in peripheral blood lymphocytes of mice through fixation and lysis and to evaluate the feasibility of applying this method to research on the radiation-related biological effects and the efficacy evaluation of radioprotective drugs.Methods:A total of 41 male C57BL/6J mice were used. First, 21 mice were randomly divided into 7 groups according to different radiation doses (0, 1, 2, 4, 6, 8, and 10 Gy) with 3 mice in each group. Blood samples were collected from the tail vein of mice at 1, 4, 8, and 24 h after irradiation and immediately fixed with formaldehyde. Red blood cells (RBC) were lysed with Triton X-100, and γ-H2AX was labeled with specific antibodies. DRAQ5 dye was used to further exclude debris and anucleate cells. The mean fluorescence intensity of γ-H2AX in lymphocyte populations was directly analyzed by flow cytometry through forward and side scatter, and dose-effect curves after irradiation were established. Then, the other 20 mice were divided into radiation alone groups and radiation combined with WR-2721 administration groups at 4 and 6 Gy, respectively, with 5 mice in each group. Blood samples were collected from the tail vein of mice at 1, 4, 8, and 24 h after irradiation to detect the average fluorescence intensity of γ-H2AX in lymphocytes, which was used to evaluate the degree of DNA damage in mice and the therapeutic effect of WR-2721.Results:The expression of γ-H2AX in peripheral blood lymphocytes of mice significantly increased with the increase of radiation doses, and reached a peak at 1-2 h and then decreased. The dose-effect relationship was significant ( R2 = 0.9914). At 24 h after 4 and 6 Gy irradiation, compared with the radiation alone groups, the average fluorescence intensity of γ-H2AX in the radiation combined with WR-2721 administration groups was lower (144.8 ± 8.0 and 109.5 ± 9.7, vs. 178.0 ± 18.5 and 136.6 ± 5.4), with statistically significant difference ( t = 3.78, 5.48, P < 0.05). The average fluorescence intensity of γ-H2AX at 24 h after irradiation was consistent with the lowest values of the three blood cell lines at 7 or 14 d after irradiation. Conclusions:The application of flow cytometry with a fixation/dissolution protocol to directly detect the mean fluorescence intensity of γ-H2AX in peripheral blood lymphocytes of mice has significant application value in radiation biology effect research, radiation protection drug screening, and efficacy evaluation.

Objective:To develop a method of employing flow cytometry to directly detect the γ-H2AX expression levels in peripheral blood lymphocytes of mice through fixation and lysis and to evaluate the feasibility of applying this method to research on the radiation-related biological effects and the efficacy evaluation of radioprotective drugs.Methods:A total of 41 male C57BL/6J mice were used. First, 21 mice were randomly divided into 7 groups according to different radiation doses (0, 1, 2, 4, 6, 8, and 10 Gy) with 3 mice in each group. Blood samples were collected from the tail vein of mice at 1, 4, 8, and 24 h after irradiation and immediately fixed with formaldehyde. Red blood cells (RBC) were lysed with Triton X-100, and γ-H2AX was labeled with specific antibodies. DRAQ5 dye was used to further exclude debris and anucleate cells. The mean fluorescence intensity of γ-H2AX in lymphocyte populations was directly analyzed by flow cytometry through forward and side scatter, and dose-effect curves after irradiation were established. Then, the other 20 mice were divided into radiation alone groups and radiation combined with WR-2721 administration groups at 4 and 6 Gy, respectively, with 5 mice in each group. Blood samples were collected from the tail vein of mice at 1, 4, 8, and 24 h after irradiation to detect the average fluorescence intensity of γ-H2AX in lymphocytes, which was used to evaluate the degree of DNA damage in mice and the therapeutic effect of WR-2721.Results:The expression of γ-H2AX in peripheral blood lymphocytes of mice significantly increased with the increase of radiation doses, and reached a peak at 1-2 h and then decreased. The dose-effect relationship was significant ( R2 = 0.9914). At 24 h after 4 and 6 Gy irradiation, compared with the radiation alone groups, the average fluorescence intensity of γ-H2AX in the radiation combined with WR-2721 administration groups was lower (144.8 ± 8.0 and 109.5 ± 9.7, vs. 178.0 ± 18.5 and 136.6 ± 5.4), with statistically significant difference ( t = 3.78, 5.48, P < 0.05). The average fluorescence intensity of γ-H2AX at 24 h after irradiation was consistent with the lowest values of the three blood cell lines at 7 or 14 d after irradiation. Conclusions:The application of flow cytometry with a fixation/dissolution protocol to directly detect the mean fluorescence intensity of γ-H2AX in peripheral blood lymphocytes of mice has significant application value in radiation biology effect research, radiation protection drug screening, and efficacy evaluation.

Objective:To compare the differences in the prevalence of mild micro-hepatic encephalopathy (MHE) among patients with cirrhosis by using the psychometric hepatic encephalopathy score (PHES) and the Stroop smartphone application (Encephal App) test.Methods:This prospective, multi-center, real-world study was initiated by the National Clinical Medical Research Center for Infectious Diseases and the Portal Hypertension Alliance and registered with International ClinicalTrials.gov (NCT05140837). 354 cases of cirrhosis were enrolled in 19 hospitals across the country. PHES (including digital connection tests A and B, digital symbol tests, trajectory drawing tests, and serial management tests) and the Stroop test were conducted in all of them. PHES was differentiated using standard diagnostic criteria established by the two studies in China and South Korea. The Stroop test was evaluated based on the criteria of the research and development team. The impact of different diagnostic standards or methods on the incidence of MHE in patients with cirrhosis was analyzed. Data between groups were differentiated using the t-test, Mann-Whitney U test, and χ2 test. A kappa test was used to compare the consistency between groups. Results:After PHES, the prevalence of MHE among 354 cases of cirrhosis was 78.53% and 15.25%, respectively, based on Chinese research standards and Korean research normal value standards. However, the prevalence of MHE was 56.78% based on the Stroop test, and the differences in pairwise comparisons among the three groups were statistically significant (kappa = -0.064, P < 0.001). Stratified analysis revealed that the MHE prevalence in three groups of patients with Child-Pugh classes A, B, and C was 74.14%, 83.33%, and 88.24%, respectively, according to the normal value standards of Chinese researchers, while the MHE prevalence rates in three groups of patients with Child-Pugh classes A, B, and C were 8.29%, 23.53%, and 38.24%, respectively, according to the normal value standards of Korean researchers. Furthermore, the prevalence rates of MHE in the three groups of patients with Child-Pugh grades A, B, and C were 52.68%, 58.82%, and 73.53%, respectively, according to the Stroop test standard. However, among the results of each diagnostic standard, the prevalence of MHE showed an increasing trend with an increasing Child-Pugh grade. Further comparison demonstrated that the scores obtained by the number connection test A and the number symbol test were consistent according to the normal value standards of the two studies in China and South Korea ( Z = -0.982, -1.702; P = 0.326, 0.089), while the other three sub-tests had significant differences ( P < 0.001). Conclusion:The prevalence rate of MHE in the cirrhotic population is high, but the prevalence of MHE obtained by using different diagnostic criteria or methods varies greatly. Therefore, in line with the current changes in demographics and disease spectrum, it is necessary to enroll a larger sample size of a healthy population as a control. Moreover, the establishment of more reliable diagnostic scoring criteria will serve as a basis for obtaining accurate MHE incidence and formulating diagnosis and treatment strategies in cirrhotic populations.

Objective:To analyze the global research hotspots in the field of pediatrics based on the Essential Science Indicators (ESI) database and explore the inspiration to domestic editors and pediatrics researchers.Methods:The journal distribution, country (region) distribution, cooperation, organization distribution, funding, publication language, hot topic words and other data of highly cited papers in the field of pediatrics in ESI database were collected and analyzed.Results:A total of 682 highly cited pediatrics papers were collected from 77 pediatrics journals included in Science Citation Index(SCI). Most of the highly cited pediatrics papers (182) were found to be published in Pediatrics.All 682 paper were published in English and frequently, characterized by multiple authors, institutions and fund support.Of 682 highly cited pediatrics papers, 435 papers were published in the United States(the first), 123 papers in England(the second) and 86 paper in Canada(the third). Novel coronavirus pneumonia, coronavirus, SARS coronavirus, autism and multiple system inflammatory syndrome are the main frontiers of global pediatric research at present.Specifically, focal pediatric system diseases mainly include respiratory system diseases, digestive system diseases, cardiovascular diseases, etc. Conclusions:ESI-based analysis of global research hotspots in the field of pediatrics provides reference materials for domestic and foreign pediatrics researchers to understand the global academic frontiers and development trends in the field of pediatrics and select topics for future scientific research.More importantly, this analysis can help domestic editors of pediatrics journals to plan topics and organize hot papers, so as to improve the academic quality and international influence of the journals.

Lycophytes and seed plants constitute the typical vascular plants. Lycophytes have been thought to have no paleo-polyploidization although the event is known to be critical for the fast expansion of seed plants. Here, genomic analyses including the homologous gene dot plot analysis detected multiple paleo-polyploidization events, with one occurring approximately 13-15 million years ago (MYA) and another about 125-142 MYA, during the evolution of the genome of Selaginella moellendorffii, a model lycophyte. In addition, comparative analysis of reconstructed ancestral genomes of lycophytes and angiosperms suggested that lycophytes were affected by more paleo-polyploidization events than seed plants. Results from the present genomic analyses indicate that paleo-polyploidization has contributed to the successful establishment of both lineages-lycophytes and seed plants-of vascular plants.
Evolution, Molecular ; Genome, Plant ; Genomics ; Phylogeny ; Polyploidy ; Selaginellaceae/genetics*

Objective To explore the related factors and clinical significance of the effect of irrigation on infant with dacryocystitis. Methods A total of 318 eyes of 262 infant were treated with dacryocystitis, and their overall curative effect was observed. Single factor correlation analysis and multivariate Logistic regression were used to analyze 8 factors related to efficacy, such as age of children, length of medical history, severity of symptoms, degree of operation of nurses, degree of parental cooperation, compliance with medication, massage therapy and treatment methods. The effect of each factor on the curative effect. Results It was found that the total effective rate was 90.56 percent of the lacrimal passage in children with 318 tear channel obstruction and dacryocystitis.Single factor analysis results show that the factors influencing the curative effect of single in treatment (χ2= 88.984, P < 0.01), symptom severity (χ2=14.185,P < 0.05) length of history (χ2=18.783, P < 0.05) difference and massage therapy (χ2=10.081, P < 0.05) was statistically significant, the three factors of multiariable Logistic regression analysis results showed that the treatment (P=0.000, OR=0.148,95％ CI 0.052- 0.419) and massage therapy (P=0.012, OR=3.390, 95％ CI 1.309- 8.777) affected infant lacrimal duct flushing out main factors influencing the efficacy of tong. Conclusions The main influencing factors are the different treatment modalities, severity of symptoms the length of medical history and massage therapy in the related factors that affect the effect of infantwith dacryocystitis.

Objective To investigate the value of serum high sensitive C reactive protein(hs-CRP) in the risk stratification and prognosis evaluation in the patients with coronary heart disease and PCI operation.Methods One hundred and sixty-three cases of coronary heart disease and undergoing PCI in this hospital from March 2013 to September 2015 were selected and performed the coronary angiography examination before operation.Then the patients were divided into 3 groups according to the hs-CRP level.The Gensini score was used to judge the blood vessel stenosis degree.The patients were followed up within six months after surgery.The major cardiovascular events were recorded and the relationship between the hs-CRP level with the number of diseased vessels,Gensini score,MACE and other coronary heart disease risk factors was analyzed.Results Hypertension and hs-CRP level had statistically significant difference among 3 groups(P＜0.05);preoperative hs-CRP level showed positive correlation with hs-cTnT level (r =0.44,P＜ 0.001),while the hs-CRP was negatively correlated with the HDL-C level (r =-0.18,P =0.003);with the Gemini score elevation,the hs-CRP level was similarly elevated,the hs-CRP levels in the patients with severe,moderate and mild vascular stenosis evaluated by Gensini's score were(17.11±9.58),(10.02±3.59),(4.11±1.62)mg/L respectively,the difference among them had statistical significance(F=37.21,P＜0.05).Within postoperative six months,the MACE occurrence rates in 3 groups were 32.1％,52.9 ％ and 59.2 ％ respectively,the differences among the three groups were statistically significant(x2 =13.523,P =0.001),and the target lesion blood supply reconstruction rate had statistically significant difference(x2 =7.522,P =0.022).The hsCRP level was an independent risk factor of MACE occurrence(OR =2.06,95 ％ CI:1.31-3.42,P=0.005).Conclusion Serum hs-CRP is an independent risk factor for cardiovascular events occurrence after PCI surgery in the patients with coronary heart disease.

The dual luciferase reporter gene system provides sensitive readout, while it relies on a constitutively-expressed control gene for readout normalization. However, most standard control reporter genes are not constitutively expressed under all conditions. Here, we report an effective method to construct a control reporter plasmid for the dual luciferase reporter gene system that would be suitable for hormone research in silkworm cell lines. First, we modified BmVgP78M, a stably-expressed constitutive promoter in silkworm cells by mutating its hormone-related element. Then, we constructed the pRL-VgP78M control reporter plasmid by replacing the SV40 promoter and chimeric intron sequences in pRL-SV40 with the BmVgP78M sequence. Finally, we confirmed that the pRL-VgP78M control reporter plasmid could be stably expressed in silkworm cell lines via cell transfection experiments, and it was unresponsive to the induction of ecdysone, juvenile hormone, or their transcription factors. We thus obtained a control reporter plasmid pRL-VgP78M that could be expressed stably and moderately in silkworm cells. It can be readily used as the control reporter plasmid of the dual luciferase reporter gene system for hormone research in silkworm cell lines. It will also provide a reference for construction of control reporter plasmids of dual luciferase reporter gene systems that are adaptable to cell lines isolated from other species.

By combining frequency modulation Chirp Z transform ion mobility spectrometers (IMS) and multi nozzle electrospray array ionization source, a method of NanoESI-Chirp Z transform ion mobility spectrometry-high performance liquid chromatography was developed for the determination of n-alkyl ammonium bromide compounds.The parameters of NanoESI-Chirp Z transform IMS such as electric field intensity, solvent composition, and solution flow rate were investigated and optimized.Subsequently, four kinds of n-alkyl ammonium bromide compounds were respectively detected by this developed Chirp Z transform method and Fourier transform method, and the obtained results were compared.The result indicated that the optimum conditions were electric field intensity of 4.5 kV, and ESI solution flow rate of 8 μL/min.Then a test mixture containing tetrabutylammonium bromide, tetrapentylammoniumbromide, tetrahexylammonium bromide, tetraheptylammonium bromide, tetranoctylammoniumbromideandtetrakis(decyl) ammonium bromide was successfully separated and determined by the HPLC-nanoESI-Chirp Z IMS method.Chirp Z transform method provided higher signal to noise ratio compared to conventional signal averaging method, and was superior to FT method in the determination of drift time.

Objective To observe the effects of herb-partition moxibustion at navel on the expressions of Bcl-2 and Bax in rats with premature ovarian failure (POF); To explore its mechanism underlying improvement of POF. Methods POF model was established by intraperitoneal injection of cyclophosphamide. Forty-eight Female Wistar rats were randomly divided into normal group, model group, moxibustion group and Western medicine group, with 12 rats in each group. Rats of moxibustion group were treated with herb-partition moxibustion at navel (Shenque, CV8), 30 min/d, once every other day; rats of Western medicine group were given intragastric dose of estradiol valerate, once a day, for 14 d. The protein and mRNA expressions of Bax and Bcl-2 in ovarian granulosa cells were detected by immunohistochemal method, Western blot and qRT-PCR, respectively.Results The weights of ovaries in the model group rats were significantly lower than those in the normal group (P<0.05). The expressions of Bcl-2 and Bax protein in ovarian tissue of each group were mainly expressed brown coloration and in the cytoplasm of cells. The range of Bax protein in granulosa cells of moxibustion group and Western medicine group was significantly less than that in model group. The range of Bcl-2 in moxibustion group and Western medicine group was significantly larger than that in model group. Compared with the normal group, the expressions of Bcl-2 protein and mRNA in the model group rats significantly decreased (P<0.05,P<0.01), and the expression of Bax protein and mRNA significantly increased (P<0.05,P<0.01). The expression level of Bcl-2 protein and mRNA was significantly higher in the moxibustion group and in the western medicine group than that in the model group (P<0.05,P<0.01). The expression level of Bax protein and mRNA in the moxibustion group was significantly lower than that in the model group (P<0.05,P<0.01). ConclusionThe herb-partition moxibustion at navel can improve the function of ovaries by up regulating the expression of Bcl-2 and down regulating the expression of Bax in POF rats model.