Novel insights into the effect of Xiaoyao san on corticosterone-induced hepatic steatosis:inhibition of glucocorticoid receptor/perilipin-2 signaling pathway

Objective:Xiaoyao san(XYS)is a classic traditional Chinese medicinal formula.It has been clinically administered to regulate liver function.However,its mechanisms in glucocorticoid-induced hepatic steatosis are unknown.This study aimed to investigate whether XYS protects against corticosterone(CORT)-induced hepatic steatosis,and to explore its mechanism.Methods:High-fat diet mice induced with hepatic steatosis by 2mg/kg CORT were administered 2.56 g/kg or 5.12 g/kg XYS daily for 7 weeks.The effects of XYS on hepatic steatosis in mice were evaluated by H&E and Oil Red O staining and by measuring their plasma lipids(triglyceride,total cholesterol,and free fatty acids).The mechanism of XYS against hepatic steatosis was investigated by network pharmacology,immunohistochemistry,western blotting,and gain-of-function/loss-offunction experiments.Results:XYS alleviated CORT-induced steatosis,decreased plasma lipids,and inhibited glucocorticoid receptor(GR)activation in the liver.Network pharmacology data indicated that XYS may have mitigated hepatic steatosis via GR which mediated adipose differentiation-related protein(ADFP).Gain-of-function/loss-of-function experiments in vitro confirmed that GR positively regulated ADFP expression.Conclusions:XYS ameliorated CORT-induced hepatic steatosis by downregulating the GR/ADFP axis and inhibiting lipid metabolism.Our studies implicate that XYS is promising as a therapy for CORT-induced hepatic steatosis,and lay the foundation for designing novel prophylactic and therapeutic strategies on CORT-induced hepatic steatosis.

Impact of co-infections with enteric pathogens on children suffering from acute diarrhea in southwest China

Background:Acute diarrhea is a global health problem,resulting in high morbidity and mortality in children.It has been suggested that enteric pathogen co-infections play an important role in gastroenteritis,but most research efforts have only focused on a small range of species belonging to a few pathogen groups.This study aimed to assess the impact of co-infections with a broad range of enteric pathogens on children aged below five years who suffer from acute diarrhea in southwest China.Method:A total of 1020 subjects(850 diarrhea cases and 170 healthy controls)were selected from four sentinel hospitals in Kunming,Yunnan province,southwest China,from June 2014 to July 2015.Stool specimens were collected to detect five virus(rotavirus group A,RVA;norovirus,NoV;Sapovirus,SaV;astrovirus,As;and adenovirus,Ad),seven bacterial(diarrheagenic Escherichia coli,DEC;non-typhoidal Salmonella,NTS;Shigella spp.;Vibrio cholera;Vibrio parahaemolyticus;Aeromonas spp.;and Plesiomonas spp.),and three protozoan(Cryptosporidium spp.,Giardia lamblia,and Blastocystis hominis,B.hominis)species using standard microbiologic and molecular methods.Data were analyzed using the partial least square regression technique and chi-square test.Results:At least one enteric pathogen was detected in 46.7%(n=397)of acute gastroenteritis cases and 13.5%(n=23)of healthy controls(χ^(2)=64.4,P<0.05).Single infection with RVA was associated with acute diarrhea(26.5%vs.5.8%,P<0.05).The prevalence of a single infection with B.hominis in diarrhea cases was higher than in healthy controls(3.1%vs.0.5%,OR=4.7,95%CI:1.01–112.0).Single infection with NoV GII was not associated with diarrhea(4.4%vs.3.5%,OR=1.2,95%CI:0.5–3.3).Single infections with bacterial species were not observed.The prevalence of co-infections with two enteric pathogens in diarrhea cases was higher than in asymptomatic children(20.1%vs.5.3%,P<0.05).RVA-NoV GII was the most common co-infection in symptomatic children(4.4%),with it aggravating the severity of diarrhea.Conclusions:Although it is clear that RVA has an overwhelming impact on diarrhea illnesses in children,co-infection with other enteric pathogens appears to also aggravate diarrhea severity.These findings should serve as evidence for public health services when planning and developing intervention programs.

HIV and intestinal parasite co-infections among a Chinese population: an immunological profile

Background:Parasite infections often result in a switch of the human body’s predominant immune reaction from T-helper 1(Th1)-type to Th2-type.Hence,parasite infections are widely expected to accelerate the progression of human immunodeficiency virus(HIV)infections to acquired immunodeficiency syndrome(AIDS).In the People’s Republic of China,both parasitic diseases and AIDS are epidemic in certain rural areas,and co-infections are relatively common.However,no population-based studies have yet investigated the frequency of HIV and parasite co-infections,and its effects on immune responses.We studied(1)the immune status of an HIV-infected population,and(2)the effect of co-infection of HIV and intestinal parasites on selected parameters of the human immune system.Methods:A total of 309 HIV-infected individuals were recruited and compared to an age-matched and sex-matched control group of 315 local HIV-negative individuals.Questionnaires were administered to all participants to obtain information on sociodemographic characteristics,sanitation habits,family income,and recent clinical manifestations.Two consecutive stool samples and 10 ml samples of venous blood were also collected from each individual for the diagnosis of parasite infections and quantitative measurements of selected cytokines and CD4+T-lymphocytes,respectively.Results:During the study period,79 HIV-infected individuals were not under highly active antiretroviral therapy(HAART)and were thus included in our analysis;the prevalence of intestinal helminth infections was 6.3%and that of protozoa was 22.8%.The most common protozoan infections were Blastocystis hominis(B.hominis)(13.9%)and Cryptosporidium spp.(10.1%).The prevalence of Cryptosporidium spp.in HIV-infected individuals was significantly higher than that in HIV negative individuals(P<0.05).Compared to the non-co-infected population,no significant difference was found for any of the measured immunological indicators(P>0.05).However,the following trends were observed:IFN-γlevels were lower,but the IL-4 level was higher,in the population co-infected with HIV and helminths.In the population co-infected with HIV and B.hominis,the IL-2 level was higher.The population co-infected with HIV and Cryptosporidium spp.had markedly lower CD4+T-lymphocyte counts.Conclusion:According to the immunologic profile, co-infection with helminths is disadvantageous to HIV-infected individuals. It was associated with a shift in the Th1/Th2 balance in the same direction as that caused by the virus itself, which might indicate an acceleration of the progress from an HIV infection to AIDS. Co-infection with Cryptosporidium spp. was not associated with a significant change in immune factors but co-infection with Cryptosporidium spp. was associated with a reduced level of CD4 + T-lymphocytes, confirming the opportunistic nature of such infections. Co-infection with B. hominis, on the other hand, was associated with an antagonistic shift in the immunological profile compared to an HIV infection.

Antibiotic resistance and molecular characterization of diarrheagenic Escherichia coli and non-typhoidal Salmonella strains isolated from infections in Southwest China

Background:Bacterial diarrhea is one of the most common causes for medical consultations,mortality and morbidity in the world.Diarrheagenic Escherichia coli(DEC)and non-typhoidal Salmonella(NTS)are major intestinal pathogens in developing countries,and the indiscriminate use of antibiotics has greatly contributed to resistant strains.Hence,the aim of the present study is to identify the antimicrobial resistance patterns and the molecular characteristics of DEC and NTS in southwest,China.Methods:1121 diarrheal patients and 319 non-diarrheal subjects across all age groups were recruited from four sentinel hospitals from June 2014 to July 2015 in Kunming City,Yunnan Province.Each stool specimen was collected to detect DEC and NTS with standard microbiological and molecular methods.Antimicrobial resistance testing was performed with the Kirby-Bauer disk diffusion method,and the standards for antimicrobial susceptibility testing complied with the Clinical and Laboratory Standards Institute(CLSI).Molecular characterization of strains was carried out using pulsed-field gel electrophoresis(PFGE).A structured questionnaire was used to record basic epidemiological data(e.g.sex,age,residence,season,etc.).Data were analyzed using Chi-square or Fisher’s exact test.Results:DEC was detected in 127(11.33%)diarrhea cases and 9(2.82%)non-diarrheal cases(χ^(2)=20.69,P<0.001,OR=4.36,95%CI:2.19-8.65),and the prevalence of NTS isolated from diarrhea cases was higher than that of non-diarrheal cases across all age groups(n=42,3.75%,n=1,0.31%,χ^(2)=10.10,P=0.002,OR=12.38,95%CI:1.70-90.29).The rates of resistance to ten antibiotics of DEC and NTS showed significant differences(χ^(2)=386.77,P<0.001;χ^(2)=191.16,P<0.001).The rates of resistance to Amoxicillin and Clavulafiate(AMC),Cephalothin(CEP),Gentamicin(GEN)and Sulfamethoxazole-Trimethoprim(SXT)of DEC isolated from diarrhea cases were higher than those of NTS isolated from diarrhea patients(37.01%vs 14.29%,χ^(2)=7.57,P=0.006;29.92%vs 11.90%,χ^(2)=5.40,P=0.02;37.01%vs 11.90%,χ^(2)=5.80,P=0.016;62.20%vs 26.19%,χ^(2)=16.44,P<0.001;respectively).Ciprofloxacin(CIP)was the most sensitive antibiotic for DEC and NTS strains isolated from diarrhea cases.Resistance rates of DEC isolates from cases and controls to more than three kinds antimicrobials(multidrug resistance,MDR)showed no significant differences(81.10%vs 88.89%,P=0.33).Pulsotype patterns of DEC strains were highly diverse;however,the pulsotype pattern of NTS strains was closely related to the serotype.The pattern of S.enteritidis was highly similar,but the S.enterica Typhimurium strain was discrete.Conclusions:Antibiotic resistance of Enterobacteriaceae is of great concern.The societal effects of antibiotic use justify strict monitoring to combat increases in antimicrobial resistance.Molecular epidemiology and systematic epidemiological investigation can provide accurate evidence for tracking the infection source.

Risk factors for Blastocystis infection in HIV/AIDS patients with highly active antiretroviral therapy in Southwest China

Background:Blastocystis is a widespread zoonotic protozoan of mammalian species,especially in HIV/AIDS individuals.The aim of this study was to analyze the prevalence and risk factors related with Blastocystis infection among HIV/AIDS patients in Southwest China.Methods:The cross-sectional study was performed in 311 HIV/AIDS cases in Tengchong City,Yunnan Province from July 2016 to March 2017.For each subject,stool specimen was collected to detect the Blastocystis,and the blood sample was used to detect HIV virus load and CD4+T cell count,in addition,structured questionnaire was used to collect the basic information and risk factors.Findings:The result showed that the detection rate of Blastocystis was 3.86%(95%CI:2.22-6.62)among HIV/AIDS patients.Both raising animal(OR=12.93,95%CI:1,54-108.36)and drinking un-boiled water(OR=8A7,95%CI:1.76-37.90)were risk factors for Blastocystis infection in HIV/AIDS individuals.In addition,the interaction of CD4^+T cell count and HIV virus load was also contribution to Blastocystis infection(P=0.007).Conclusions:A high prevalence of Blastocystis infection was found in HIV/AIDS patients in Tengchong.Poor hygienic habits,the interaction of HIV virus load and CD4^+T cell count were identified as main risk factors for infection.These results will help us to develop efficient control strategies to intervene with and prevent the occurrence of Blastocystis among HIV-infected individuals.

Trypanosoma brucei rhodesiense infection in a Chinese traveler returning from the Serengeti National Park in Tanzania

Background:Human African trypanosomiasis(HAT)is one of the most complex parasitic diseases known to humankind.It usually occurs in endemic areas in Africa,but is occasionally detected in returning travelers and migrants in non-endemic countries.Case presentation:In August 2017,a case of HAT was diagnosed in China in a traveler returning from the Masai Mara area in Kenya and the Serengeti area in Tanzania.The traveler visited Africa from 23 July to 5 August,2017.Upon return to China,she developed a fever(on 8 August),and Trypanosoma brucei rhodesiense infection was confirmed by laboratory tests(on 14 August)including observation of parasites in blood films and by polymerase chain reaction.She was treated with pentamidine followed by suramin,and recovered 1 month later.Conclusions:This is the first imported rhodesiense HAT case reported in China.This case alerts clinical and public health workers to be aware of HAT in travelers,and expatriates and migrants who have visited at-risk areas in Africa.

A neglected risk for sparganosis:eating live tadpoles in central China

A 29-year-old farmer from central China was sent into the Emergency Department of the Affiliated Hospital of Zhengzhou University.He had a 15-day history of persistent high fever,abdominal distention and pain.The patient was clinically diagnosed as appendicitis and peritonitis,and treated with antibiotics in a local hospital,did not improve.On exploratory laparotomy,the appendicular perforation and peritonitis were seen;appendicectomy were performed,and antibiotics were given.However,high fever and abdominal pain still persisted;intestinal adhesion and obstruction,ascites appeared.He was given the“critically ill notice”.He had eosinophilia(12.95%)and the history of eating live frog tadpoles for treating his cutaneous pruritus 3 days before onset of the disease.Serum anti-sparganum antibodies assayed by ELISA were positive.This patient has hospitalized for one and half months and spend more than US$12000.This patient was primarily diagnosed as visceral sparganosis,and cured with praziquantel.Sparganosis is one neglected but important parasitic zoonosis of poverty.Human infections were mainly acquired by eating raw or uncooked meat of frogs and snakes infected with plerocercoids,using frog or snake flesh as poultices,or drinking raw water contaminated with infected copepods.However,sparganosis caused by ingestion of live tadpoles are emerging in central China.Our surveys showed that 11.93%of tadpoles in Henan province are infected with plerocercoids.Eating live tadpoles is a high risk for sparganum infection.The comprehensive public health education should be carried out for people in endemic areas and the bad habit of eating live tadpoles must be discouraged.

Xiaoyao San,a Chinese herbal formula,ameliorates depression-like behavior in mice through the AdipoR1/AMPK/ACC pathway in hypothalamus

Objective Depression and metabolic disorders have overlapping psychosocial and pathophysiological causes.Current research is focused on the possible role of adiponectin in regulating common biological mechanisms.Xiaoyao San(XYS),a classic Chinese medicine compound,has been widely used in the treatment of depression and can alleviate metabolic disorders such as lipid or glucose metabolism disorders.However,the ability of XYS to ameliorate depression-like behavior as well as metabolic dysfunction in mice and the underlying mechanisms are unclear.Methods An in vivo animal model of depression was established by chronic social defeat stress(CSDS).XYS and fluoxetine were administered by gavage to the drug intervention group.Depression-like behaviors were analyzed by the social interaction test,open field test,forced swim test,and elevated plus maze test.Glucose levels were measured using the oral glucose tolerance test.The involvement of certain molecules was validated by immunofluorescence,histopathology,and Western blotting.In vitro,hypothalamic primary neurons were exposed to high glucose to induce neuronal damage,and the neuroprotective effect of XYS was evaluated by cell counting kit-8 assay.Immunofluorescence and Western blotting were used to evaluate the influences of XYS on adiponectin receptor 1(AdipoR1),adenosine 5’-monophosphate-activated protein kinase(AMPK),acetyl-coenzyme A carboxylase(ACC)and other related proteins.Results XYS ameliorated CSDS-induced depression-like behaviors and glucose tolerance impairment in mice and increased the level of serum adiponectin.XYS also restored Nissl bodies in hypothalamic neurons in mice that exhibited depression-like behaviors and decreased the degree of neuronal morphological damage.In vivo and in vitro studies indicated that XYS increased the expression of AdipoR1 in hypothalamic neurons.Conclusion Adiponectin may be a key regulator linking depression and metabolic disorders;regulation of the hypothalamic AdipoR1/AMPK/ACC pathway plays an important role in treatment of depression by XYS.

Profiling B and T cell immune responses to co-infection of Mycobacterium tuberculosis and hookworm in humans

Background:Humoral and cellular immune responses play protective roles against Mycobacterium tuberculosis(MTB)infection.However,hookworm infection decreases the immune response to hookworm and bystander antigens.Currently,immune responses to co-infection of MTB and hookworm are still unknown,although co-infection has been one of the public health problems in co-endemic areas of pulmonary tuberculosis(PTB)and hookworm disease.Therefore,it is essential to evaluate B and T cell immune responses to the co-infection.Methods:Seventeen PTB cases co-infected with hookworm,26 PTB cases,15 patients with hookworm infection,and 24 healthy controls without PTB or hookworm infection were enrolled in the study.Expressions of CD3,CD4,CD8,CD10,CD19,CD20,CD21,CD25,CD27,CD38,FoxP3,and PD-1 were assessed on B and T cell subsets using multicolor flow cytometry.Results:For the B cell(CD19+)subsets,naïve B cells(CD10−CD27−CD21+CD20+),plasma cells(CD10−CD27+CD21−CD20−),and tissue-like memory B cells(CD10−CD27−CD21−CD20+)had higher proportions,whilst resting memory B cells(CD10−CD27+CD21+CD20+)had lower proportions in the group co-infected with MTB and hookworm as compared to other groups.Frequencies of activated memory B cells(CD10−CD27+CD21−CD20+)did not differ among the four groups.For the T cell(CD3+)subsets,frequencies of regulatory T cells(CD4+CD25+Foxp3+)and exhausted CD4+and CD8+T cells(CD4+PD-1+and CD8+PD-1+)were higher,and frequencies of activated CD4+and CD8+T cells(CD4+CD38+and CD8+CD38+)were lower in the co-infected group as compared to the other groups.Conclusion:The change patterns of the cell profile of circulating lymphocytes were indentified in human co-infection of MTB and hookworm,which might indicate that the humoral and cellular immune responses are more suppressed.

Co-infection of HIV and parasites in China: Results from an epidemiological survey in rural areas of Fuyang city, Anhui province, China

The aim of this study was to investigate epidemiological characteristics of co-infection of HIV and intestinal parasites in a HIV/AIDS highly endemic area in China.A cross-sectional survey was carried out in two villages where HIV/AIDS prevalence in residents was over 1%.Stool samples of all residents in the two targeted villages were examined for the presence of intestinal parasites.Blood examination was performed for the HIV infection detection and anemia test.A questionnaire survey was carried out in all residents participating in the study.A total of 769 individuals were enrolled in the investigation,720 of whom were involved in stool examination of intestinal parasites.The infection rates of parasites in the residents of the targeted villages were as follows:0.56%for Ascaris lumbricoides,4.03%for Hookworm,0.28%for Trichuris trichiura,0.42%for Clonorchis sinensis,21.39%for Blastocystis hominis,3.89%for Giardia lamblia,1.67%for Entamoeba spp.,4.44%for Cryptosporidium spp.,and no infection for Strongyloides stercoralis.The overall infection rate of intestinal worms was 4.72%,intestinal parasite infection rate was 24.31%,the anemia prevalence rate was 34.68%,the co-infection rate of HIV and intestinal helminthes 2.17%,the co-infection rate of HIV and intestinal protozoa 28.26%,of which the co-infection rates of HIV and Blastocystis hominis,and HIV and Cryptosporidium spp.were 19.57%and 13.04%,respectively.A significant difference of Cryptosporidium spp.infection rate was found between HIV-positive group(13.04%)and HIV-negative group(4.70%)(P<0.05).Higher prevalence of anemia(34.68%)occurred in the study villages,particularly in children(>50%)and women(>39%).It was concluded that the infection rate of intestinal protozoa in the residents was higher than that of intestinal helminthes in the local setting,and a same pattern appeared in the co-infection rate of HIV and parasites.An interestingfinding is that the infection rate of Cryptosporidium spp.among the HIV-positives was significantly higher than that in the HIV-negatives,and children and women had higher anemia prevalence.

Urgent needs in fostering neglected tropical diseases(NTDs)laboratory capacity in WHO Western Pacific Region:results from the external quality assessment on NTDs diagnosis in 2012-2015

Background:Neglected tropical diseases(NTDs)are a heterogeneous group of mainly chronic,debilitating and often stigmatizing diseases that largely affects low-income and politically marginalized populations,causing a large burden of public health,social and economies in the NTDs endemic countries.NTDs are caused by infections with a range of pathogen,including bacteria,parasites,protozoa and viruses.The accurate diagnosis of NTDs is important for reducing morbidity,preventing mortality and for monitoring of control programs.External Quality Assessment(EQA),a component of laboratory quality assurance,aims to assess the performance of participating laboratories in detecting parasitic infections.The aim of this paper is to report the findings and put forward the recommendations on capacity build from the EQA results of participating NTDs laboratories in selected countries in the WHO Western Pacific Region from 2012 to 2015.Methods:Reference or public health laboratories at national level working on NTDs in 6 countries participated in EQAs organized by the National Institute of Parasitic Diseases(NIPD)of Chinese Center for Disease Control and Prevention(CDC)based in Shanghai,China.Two representatives of each participating laboratory were invited to NIPD to detect NTDs’parasitic infections using the same prepared samples for serological tests(IHA and ELISA)and helminth eggs’morphological tests(Direct smear and Kato-Katz).All of the results were scored and analyzed by using SPSS statistics 19.0 software.Results:The percentage of participants who had EQA score≥60 during 2012-2015 for direct smear test were 80.00%(2012),71.43%(2013),100%(2014)and 75.00%(2015),whereas for Kato-Katz test were 80.00%(2012),57.14%(2013),100%(2014)and 37.50%(2015),respectively.The detection rate of helminth eggs varied in different species,with Ascaris lumbricoides being the highest at 94.07%in average.All laboratories did very well with ELISA tests as shown by the high scores in all four years except Lab A in the first and last EQA.For the positive or negative judgments of serum samples,the total coincidence rates of ELISA between 2012 and 2015 were 90.00%,99.29%,94.29%and 98.75%,respectively.While the total coincidence rates of IHA were respectively 100%,95.00%,90.00%and 97.50%.However,detecting low levels of serum antibody remained problematic for IHA when the titres of samples were taken into consideration.Conclusion:This study demonstrate that EQA scheme have been beneficial to the participating laboratories.The EQA programme identifies certain deficiencies which were needed to overcome and improved the laboratories’performance in helminthiasis diagnosis.However,further optimization of accuracy and uniformity in NTDs diagnosis remains a big challenge.

Cross-strait parasitological research priorities arrived at by historical tracking and advanced dialogue

To further enhance dialogue and promote cross-strait cooperation in the prevention and control of parasitic diseases,this paper reviewed the progress and current challenges in the cross-strait control and research of parasitic infections,based on three cross-strait meetings on parasitological research in the last decade.The major outcome of the 3rd Meeting of Cross-Strait Parasitological Research held in April 2013 was identifying the research priorities for parasitological research.