This study established a droplet digital PCR(ddPCR)EvaGreen assay and probe methods for Schistosoma japonicum detection,and evaluated their application in detecting early infections in the S.japonicum host oncomelania and mice.Primers and corresponding probes for both ddPCR methods were designed and synthesized,and plasmids containing target sequences were constructed.The sensitivity of the two methods was tested through detection of the corresponding plasmids,and infectious and mixed oncomelania genomic DNA.Their specificity was evaluated by the detection of genomic DNA of negative oncomelania,Schistosoma mansoni,Clonorchis sinensis,Spirometra mansoni,and S.japonicum(as a positive control).The ddPCR probe method was evaluated by detection of early infection of oncomelania exposed tomiracidium with various ratios and incubation times,and the early migration and distribution of cercaria or schistosomula in mouse hosts infected with 200 cercaria via abdominal skin contact.According to standard curves constructed through the detection of plasmid serial dilutions,the regression equation for the EvaGreen assay was y=-0.839 9x+7.050 9,with a correlation coefficient R2=0.988 1,and the regression equation for the probe method was y=-1.047 5x+7.255 1,with a correlation coefficient R2=0.999 8.The lowest limit of plasmid detection with the probe method was between 38.94 cp/μL and 194.74 cp/μL.Both methods successfully detected positive reactions in the genomic DNA samples of infectious oncomelania at concentrations above 0.002 ng/μL and in the genomic DNA of each group of oncomelania mixtures.No significant differences between probe methods were observed in the detection values in the control group and the genomic DNA of negative oncomelania,S.mansoni,C.sinensis,and S.mansoni.However,the detection value of genomic DNA of negative oncomelania(291 ng/μL)with the EvaGreen assay was(20.3±4.39)cp/μL,a value significantly higher than the(1.5±0.1)cp/μL observed in the control group.For detection of early infection in oncomelania,the probe method detected Schistosoma japonicum DNA after 30 s incubation at room temperature with a≥5∶1 ratio of miracidium to oncomelania;the detection value peaked after a short time(5 min),and the peak value showed a fold increase similar to the increase in the miracidium to oncomelania ratio.Detection of early stage infection in mice with the probe method revealed that the schistosomula entered the lungs on day 2 and the liver on day 4,and continually migrated within the organs with abundant blood supply(spleen,kidney,and brain)in the first 9 days;moreover,a tendency toward ectopic parasitism was observed in the heart and pancreas on day 9,and a constantly negative control level was observed in the testes.The ddPCR probe method was more accurate and specific than the EvaGreen assay in the detection of plasmids,and infectious and mixed oncomelania,and the latter showed non-specific reactions in negative oncomelania detection.In a practical application,the probe method was demonstrated to be sensitive,to effectively reflect the early infection of oncomelania,and to reveal schistosomula migration and distribution in multiple organs of infected mice.

This study established a droplet digital PCR(ddPCR)EvaGreen assay and probe methods for Schistosoma japonicum detection,and evaluated their application in detecting early infections in the S.japonicum host oncomelania and mice.Primers and corresponding probes for both ddPCR methods were designed and synthesized,and plasmids containing target sequences were constructed.The sensitivity of the two methods was tested through detection of the corresponding plasmids,and infectious and mixed oncomelania genomic DNA.Their specificity was evaluated by the detection of genomic DNA of negative oncomelania,Schistosoma mansoni,Clonorchis sinensis,Spirometra mansoni,and S.japonicum(as a positive control).The ddPCR probe method was evaluated by detection of early infection of oncomelania exposed tomiracidium with various ratios and incubation times,and the early migration and distribution of cercaria or schistosomula in mouse hosts infected with 200 cercaria via abdominal skin contact.According to standard curves constructed through the detection of plasmid serial dilutions,the regression equation for the EvaGreen assay was y=-0.839 9x+7.050 9,with a correlation coefficient R2=0.988 1,and the regression equation for the probe method was y=-1.047 5x+7.255 1,with a correlation coefficient R2=0.999 8.The lowest limit of plasmid detection with the probe method was between 38.94 cp/μL and 194.74 cp/μL.Both methods successfully detected positive reactions in the genomic DNA samples of infectious oncomelania at concentrations above 0.002 ng/μL and in the genomic DNA of each group of oncomelania mixtures.No significant differences between probe methods were observed in the detection values in the control group and the genomic DNA of negative oncomelania,S.mansoni,C.sinensis,and S.mansoni.However,the detection value of genomic DNA of negative oncomelania(291 ng/μL)with the EvaGreen assay was(20.3±4.39)cp/μL,a value significantly higher than the(1.5±0.1)cp/μL observed in the control group.For detection of early infection in oncomelania,the probe method detected Schistosoma japonicum DNA after 30 s incubation at room temperature with a≥5∶1 ratio of miracidium to oncomelania;the detection value peaked after a short time(5 min),and the peak value showed a fold increase similar to the increase in the miracidium to oncomelania ratio.Detection of early stage infection in mice with the probe method revealed that the schistosomula entered the lungs on day 2 and the liver on day 4,and continually migrated within the organs with abundant blood supply(spleen,kidney,and brain)in the first 9 days;moreover,a tendency toward ectopic parasitism was observed in the heart and pancreas on day 9,and a constantly negative control level was observed in the testes.The ddPCR probe method was more accurate and specific than the EvaGreen assay in the detection of plasmids,and infectious and mixed oncomelania,and the latter showed non-specific reactions in negative oncomelania detection.In a practical application,the probe method was demonstrated to be sensitive,to effectively reflect the early infection of oncomelania,and to reveal schistosomula migration and distribution in multiple organs of infected mice.

AIM To investigate the effects of the Chinese medicine of Huangqi Jianzhong Decoction on excessive consumption of white adipose tissue (WAT) in a rat model of cardiac cachexia and the role of BMAL1/REV-ERBα circadian loop in WAT.METHODS Forty Dahl salt-sensitive rats were randomly divided into the control group,the normal circadian rhythm group,the normal circadian rhythm+Chinese medicine group,the mass recorded;their pathomorphological changes of WAT observed using HE and oil red staining;their UCP1 expression in WAT detected by immunohistochemistry;and their protein expression of HSL,PKA,p-PKA,BMAL1 and REV-ERBα in WAT detected by Western blot.RESULTS Compared with the control group,the normal rhythm group showed decreased rat cardiac function and body weight (P<0.05);decreased positive area of oil red staining in WAT and the inner diameter of adipocytes (P<0.01);increased positive area of UCP1 protein (P<0.01),and increased expressions of HSL,PKA,p-PKA and REV-ERBαprotein in WAT as well (P<0.01).Compared with the normal rhythm group,the disrupted circadian rhythm group displayed decreased positive area of oil red staining of WAT and the inner diameter of adipocytes (P<0.05),increased expressions of HSL,PKA,p-PKA and REV-ERBαin WAT (P<0.05),and decreased BMAL1 protein expression (P<0.05).Upon both groups of normal rhythm and disrupted circadian rhythm,the intervention of Huangqi Jianzhong Decoction increased the positive area of oil red staining in WAT and the inner diameter of adipocytes (P<0.05),decreased the positive area of UCP1 protein (P<0.01),decreased the expressions of HSL,PKA,p-PKA and REV-ERBα protein in WAT (P<0.05),and increased the expression of BMAL1 protein (P<0.05).CONCLUSION Huangqi Jianzhong Decoction may alleviate the excessive WAT depletion in the rat model of cardiac cachexia through reduction in adipose tissue dialysis and browning of white adipocytes via regulation of the circadian rhythm genes of BMAL1/REV-ERBα.

AIM To investigate the effects of the Chinese medicine of Huangqi Jianzhong Decoction on excessive consumption of white adipose tissue (WAT) in a rat model of cardiac cachexia and the role of BMAL1/REV-ERBα circadian loop in WAT.METHODS Forty Dahl salt-sensitive rats were randomly divided into the control group,the normal circadian rhythm group,the normal circadian rhythm+Chinese medicine group,the mass recorded;their pathomorphological changes of WAT observed using HE and oil red staining;their UCP1 expression in WAT detected by immunohistochemistry;and their protein expression of HSL,PKA,p-PKA,BMAL1 and REV-ERBα in WAT detected by Western blot.RESULTS Compared with the control group,the normal rhythm group showed decreased rat cardiac function and body weight (P<0.05);decreased positive area of oil red staining in WAT and the inner diameter of adipocytes (P<0.01);increased positive area of UCP1 protein (P<0.01),and increased expressions of HSL,PKA,p-PKA and REV-ERBαprotein in WAT as well (P<0.01).Compared with the normal rhythm group,the disrupted circadian rhythm group displayed decreased positive area of oil red staining of WAT and the inner diameter of adipocytes (P<0.05),increased expressions of HSL,PKA,p-PKA and REV-ERBαin WAT (P<0.05),and decreased BMAL1 protein expression (P<0.05).Upon both groups of normal rhythm and disrupted circadian rhythm,the intervention of Huangqi Jianzhong Decoction increased the positive area of oil red staining in WAT and the inner diameter of adipocytes (P<0.05),decreased the positive area of UCP1 protein (P<0.01),decreased the expressions of HSL,PKA,p-PKA and REV-ERBα protein in WAT (P<0.05),and increased the expression of BMAL1 protein (P<0.05).CONCLUSION Huangqi Jianzhong Decoction may alleviate the excessive WAT depletion in the rat model of cardiac cachexia through reduction in adipose tissue dialysis and browning of white adipocytes via regulation of the circadian rhythm genes of BMAL1/REV-ERBα.

Abstract The popularity of electronic devices helps the spread of the flow experience. Flow experience affects many aspects of life for adolescents as for its double articulation. In order to provide suggestions for the improvement in adolescent mental health, this paper reviews flow experience and adolescent mental health in different situations. In learning situation, the state of flow makes a difference in students self confidence, learning experience, positive affect in school, positivity, self esteem and self efficacy, etc., at the same time, it is beneficial to relieve academic pressure and academic procrastination. In entertainment situation, the state of flow experience directly leads to addiction, and indirectly affects online game addiction by the mediating roles of self control and cognitive bias.

OBJECTIVE: Arsenic (As) and fluoride (F) are two of the most common elements contaminating groundwater resources. A growing number of studies have found that As and F can cause neurotoxicity in infants and children, leading to cognitive, learning, and memory impairments. However, early biomarkers of learning and memory impairment induced by As and/or F remain unclear. In the present study, the mechanisms by which As and/or F cause learning memory impairment are explored at the multi-omics level (microbiome and metabolome). METHODS: We stablished an SD rats model exposed to arsenic and/or fluoride from intrauterine to adult period. RESULTS: Arsenic and/fluoride exposed groups showed reduced neurobehavioral performance and lesions in the hippocampal CA1 region. 16S rRNA gene sequencing revealed that As and/or F exposure significantly altered the composition and diversity of the gut microbiome，featuring the Lachnospiraceae_NK4A136_group, Ruminococcus_1, Prevotellaceae_NK3B31_group, [Eubacterium]_xylanophilum_group. Metabolome analysis showed that As and/or F-induced learning and memory impairment may be related to tryptophan, lipoic acid, glutamate, gamma-aminobutyric acidergic (GABAergic) synapse, and arachidonic acid (AA) metabolism. The gut microbiota, metabolites, and learning memory indicators were significantly correlated. CONCLUSION Learning memory impairment triggered by As and/or F exposure may be mediated by different gut microbes and their associated metabolites.
Rats ; Animals ; Arsenic/toxicity* ; Fluorides ; RNA, Ribosomal, 16S/genetics* ; Rats, Sprague-Dawley ; Metabolome ; Microbiota

Objective: This cross-sectional investigation aimed to determine the incidence, clinical characteristics, prognosis, and related risk factors of olfactory and gustatory dysfunctions related to infection with the SARS-CoV-2 Omicron strain in mainland China. Methods: Data of patients with SARS-CoV-2 from December 28, 2022, to February 21, 2023, were collected through online and offline questionnaires from 45 tertiary hospitals and one center for disease control and prevention in mainland China. The questionnaire included demographic information, previous health history, smoking and alcohol drinking, SARS-CoV-2 vaccination, olfactory and gustatory function before and after infection, other symptoms after infection, as well as the duration and improvement of olfactory and gustatory dysfunction. The self-reported olfactory and gustatory functions of patients were evaluated using the Olfactory VAS scale and Gustatory VAS scale. Results: A total of 35 566 valid questionnaires were obtained, revealing a high incidence of olfactory and taste dysfunctions related to infection with the SARS-CoV-2 Omicron strain (67.75%). Females(χ2=367.013, P<0.001) and young people(χ2=120.210, P<0.001) were more likely to develop these dysfunctions. Gender(OR=1.564, 95%CI: 1.487-1.645), SARS-CoV-2 vaccination status (OR=1.334, 95%CI: 1.164-1.530), oral health status (OR=0.881, 95%CI: 0.839-0.926), smoking history (OR=1.152, 95%CI=1.080-1.229), and drinking history (OR=0.854, 95%CI: 0.785-0.928) were correlated with the occurrence of olfactory and taste dysfunctions related to SARS-CoV-2(above P<0.001). 44.62% (4 391/9 840) of the patients who had not recovered their sense of smell and taste also suffered from nasal congestion, runny nose, and 32.62% (3 210/9 840) suffered from dry mouth and sore throat. The improvement of olfactory and taste functions was correlated with the persistence of accompanying symptoms(χ2=10.873, P=0.001). The average score of olfactory and taste VAS scale was 8.41 and 8.51 respectively before SARS-CoV-2 infection, but decreased to3.69 and 4.29 respectively after SARS-CoV-2 infection, and recovered to 5.83and 6.55 respectively at the time of the survey. The median duration of olfactory and gustatory dysfunctions was 15 days and 12 days, respectively, with 0.5% (121/24 096) of patients experiencing these dysfunctions for more than 28 days. The overall self-reported improvement rate of smell and taste dysfunctions was 59.16% (14 256/24 096). Gender(OR=0.893, 95%CI: 0.839-0.951), SARS-CoV-2 vaccination status (OR=1.334, 95%CI: 1.164-1.530), history of head and facial trauma(OR=1.180, 95%CI: 1.036-1.344, P=0.013), nose (OR=1.104, 95%CI: 1.042-1.171, P=0.001) and oral (OR=1.162, 95%CI: 1.096-1.233) health status, smoking history(OR=0.765, 95%CI: 0.709-0.825), and the persistence of accompanying symptoms (OR=0.359, 95%CI: 0.332-0.388) were correlated with the recovery of olfactory and taste dysfunctions related to SARS-CoV-2 (above P<0.001 except for the indicated values). Conclusion: The incidence of olfactory and taste dysfunctions related to infection with the SARS-CoV-2 Omicron strain is high in mainland China, with females and young people more likely to develop these dysfunctions. Active and effective intervention measures may be required for cases that persist for a long time. The recovery of olfactory and taste functions is influenced by several factors, including gender, SARS-CoV-2 vaccination status, history of head and facial trauma, nasal and oral health status, smoking history, and persistence of accompanying symptoms.
Female ; Humans ; Adolescent ; SARS-CoV-2 ; Smell ; COVID-19/complications* ; Cross-Sectional Studies ; COVID-19 Vaccines ; Incidence ; Olfaction Disorders/etiology* ; Taste Disorders/etiology* ; Prognosis

Objective:To retrospectively analyze the early mortality and related risk factors in adult patients with maintenance hemodialysis (MHD).Methods:Adult MHD patients from 2008 to 2018 were enrolled and divided into training data group and validation data group. In training data group, multivariate logistic regression was used to analyze the risk factors of early death within 120 days after hemodialysis and establish a prediction model. The receiver operating characteristic (ROC) curve was applied to evaluate the prediction ability of the model.Results:A total of 4 885 patients were included. The cumulative mortality within 120 days was 20.97/100 person years, and that within 365 days was 12.25/100 person years. A total of 3 603 patients in the training data group were analyzed. The following risk factors were correlated with early mortality (all P<0.05), including age at start of dialysis over 60 years old ( OR=1.792), non-chronic glomerulonephritis ( OR=2.214), cardio-cerebrovascular disease ( OR=2.695), plasma albumin less than 35 g/L ( OR=1.358), platelet count less than 120×10 9/L ( OR=2.194), serum creatinine less than 600 μmol/L ( OR=1.652), blood urea nitrogen over 30 mmol/L ( OR=1.887), blood phosphorus less than 1.13 mmol/L ( OR=1.783), pulse pressure over 55 mmHg(1 mmHg=0.133 kPa) ( OR=1.656), low density lipoprotein less than 1.5 mmol/L ( OR=1.873), and blood calcium over 2.5 mmol/L ( OR=1.876). Risk prediction model was established. The other 1 282 cases in the validation data group were verified. The area under ROC curve was 0.810, with sensitivity 85.7%, and specificity 62.5%. Conclusion:The mortality rate of adult MHD patients within 120 days after dialysis is high. The established prediction model can effectively predict the risk of early death.

Objective: To evaluate the effect of transcutaneous electrical acupoint stimulation (TEAS) on the pregnancy outcomes of in vitro fertilization-embryo transfer (IVF-ET) based on the available clinical evidence. METHODS: We searched PubMed, MEDLINE, EMBASE, Cochrane Library, CNKI, VIP, CBM and Wanfang Database up to February 2021 for published randomized controlled trials (RCT) relevant to TEAS for the improvement of the pregnancy outcomes of IVF-ET. We performed literature screening, data extraction and quality evaluation according to the inclusion and exclusion criteria, followed by a meta-analysis with the RevMan 5.3 software. RESULTS: A total of 2 206 cases of IVF-ET from 9 RCTs were included, 1 018 in the TEAS group and 1 188 in the control. The clinical pregnancy rate was significantly higher in the TEAS than in the mock TEAS and non-TEAS control groups (RR = 1.85, 95% CI: 1.42－2.42, P < 0.001; RR = 1.23, 95% CI: 1.10－1.39, P = 0.0004), and so was it before and after oocyte retrieval (RR = 1.50, 95% CI: 1.03－2.17, P = 0.03; RR = 1.47, 95% CI: 1.12－1.92, P = 0.005). The TEAS group also showed dramatically improved embryo implantation rate (RR = 1.49, 95% CI: 1.24－1.79, P < 0.0001) and live birth rate (RR = 1.44, 95% CI: 1.04－1.98, P = 0.03) compared with the control. CONCLUSIONS As a safe and non-invasive treatment, TEAS can significantly improve the pregnancy outcomes of IVF-ET, with definite effectiveness. /.
Acupuncture Points ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Pregnancy ; Pregnancy Outcome ; Randomized Controlled Trials as Topic

Objective:To study the effect of physical therapy and heel sound feedback on lower limbs motor function, mobility and activities of daily living (ADL) for stroke patients based on International Classification of Functioning, Disability and Health (ICF) core set. Methods:From April, 2018 to May, 2020, 113 stroke patients with motor dysfunction were divided into ischemia group (n = 67) and hemorrhagia group (n = 46) according to the cause of stroke. They received physical therapy for lower limbs and heel sound feedback for eight weeks, and assessed with ICF core set for stroke-gait, Fugl-Meyer Assessment-Lower Extremities (FMA-LE), Timed 'Up and Go' Test (TUGT), and modified Barthel index (MBI) before and after intervention. Results:The main effect of time was significant for qualifiers of ICF core set for stroke-gait, the scores of FMA-LE and MBI, and TUGT time (F > 100.59, P < 0.001), and it improved time by time as Post Hoc test. The main effect of groups was not significant (F < 2.29, P > 0.05), nor as Post Hoc test. The interactive effect between time and groups was significant for TUGT time (F = 6.45, P < 0.01), perhaps improved more in the hemorrhagia group, however, the interactive effect was not significant for the others. Conclusion:Physical therapy and heel sound feedback can improve motor function of lower limb, mobility and ADL for stroke patients.