Objective To study and compare the oral microbiota and metabolites of children with Henoch Schonlein purpura(HSP)to identify specific microbiota and metabolites related to this disease and elucidate the pathogenesis of HSP.Methods Three groups of qualified subjects were included,including 20 in the HSP group,20 in the HSP nephritis(HSPN)group,and 20 in the control group.Perform high-throughput 16S rRNA sequencing and metabolic profiling of saliva from each group to analyze the correlation between differential microbiota and differ-ential metabolites.Results(1)Compared with the control group,there was a significant difference in richness and diversity in the HSPN group(P＜0.05).At the same time,there was no significant difference in richness and diver-sity in the HSP group(P＞0.05).Compared with the HSP group,the abundance,and diversity of the HSPN group were significantly increased(P＜0.05).At the genus level,the proportion of Streptococcus in each group is the high-est.Compared with the control group,there was no significant correlation between the HSP group and the genus of bacteria.In contrast,the HSPN group showed a significant increase in the genera of Pseudomonas and Parabacteroi-des(P＜0.05).Compared with the HSP group,the abundance of Pseudomonas and Parabacteroides in the HSPN group was significantly increased(P＜0.05).(2)Compared with the control group,the HSPN group had 12 differen-tial metabolites involving nine metabolic pathways,such as phenylalanine metabolism;There was no significant dif-ference in metabolites and no metabolic pathway in the HSP group.Compared with the HSP group,the HSPN group has 15 differential metabolites involving nine metabolic pathways,such as phenylalanine metabolism.(3)In the HSPN and control groups,Pseudomonas and Parabacteroides negatively correlated with Phenylalanine metabolic pathway products.In the HSPN and HSP groups,Pseudomonas,Parabacteroides,and Phenylalanine metabolic path-way products were negatively correlated.The metabolites involved in phenylalanine metabolism in the oral cavity are 2-hydroxycinnamic acid,Phenylpyruvic acid,and N-acetyl-L-phenylalanine.Conclusion There is a significant dif-ference between HSPN and HSP children and healthy children.Streptococcus,Pseudomonas,and Parabacteroides may be one of the trigger factors of HSPN,and Phenylalanine metabolism may be one of the pathways in the patho-genesis of HSPN.Children with HSPN have a more pronounced imbalance in oral microbiota and greater differences in metabolic products than children with HSP.

Objective:To identify the phosphodiesterase (PDE) activity of the gene product encoded by LA_RS06960 of Leptospira interrogans ( L. interrogans), and analyze whether dinucleotides that can be degraded by PDE can activate macrophages to express innate immune factors. Methods:The LA_RS06960 gene in L. interrogans strain 56601 was amplified by PCR, and the prokaryotic expression system was constructed for the protein expression. The expressed rPDE was purified by Ni-NTA affinity chromatography. High performance liquid chromatography (HPLC) was used to measure the degration of c-di-AMP or 5′-pApA to AMP by rPDE. Real-time fluorescent quantitative RT-PCR (qRT-PCR) was used to detect the changes in the expression of target genes in Leptospira or THP-1 cells associated with innate immune factors during infection. qRT-PCR and ELISA were used to detect the changes in the expression and secretion level of the innate immune factors in macrophages treated with bacterial dinucleotide. Results:The prokaryotic expression system for LA_RS06960 gene of L. interrogans was constracted successfully, and the purified rPDE could degrate 5′-pApA and c-di-AMP into AMP in vitro. The mRNA level of leptospiral LA_RS06960 gene was significantly down-regulated, while IFN-β, TNF-α, IL-6 and IL-1β encoding genes of macrophages were significantly up-regulated during infection. The mRNA level or the secretion level of IFN-β and IL-6 of macrophages were increased after treated with the bacterial dinucleotide substrate of PDE. Conclusions:PDE encoded by LA_RS06960 gene has phosphodiesterase activity, and the bacterial dinucleotide substrate of the PDE could activate the innate immune response of macrophages.

Purpose: To intelligently diagnose whether there is bladder outlet obstruction (BOO) in female with decent detrusor contraction ability by focusing on urodynamic study (UDS) data. Materials and Methods: We retrospectively reviewed the UDS data of female patients during urination. Eleven easily accessible urinary flow indicators were calculated according to the UDS data of each patient during voiding period. Eight diagnosis models based on back propagation neural network with different input feature combination were constructed by analyzing the correlations between indicators and lower urinary tract dysfunction labels. Subsequently, the stability of diagnostic models was evaluated by five-fold cross-validation based on training data, while the performance was compared on test dataset. Results: UDS data from 134 female patients with a median age of 51 years (range, 27–78 years) were selected for our study.Among them, 66 patients suffered BOO and the remaining were normal. Applying the 5-fold cross-validation method, the model with the best performance achieved an area under the receiver operating characteristic curve (AUC) value of 0.949±0.060 using 9 UDS input features. The accuracy, sensitivity, and specificity for BOO diagnosis model in the testing process are 94.4%, 100%, and 89.3%, respectively. Conclusions The 9 significant indicators in UDS were employed to construct a diagnostic model of female BOO based on machine learning algorithm, which performs preferable classification accuracy and stability.

Objective:To investigate whether KtrA was a binding protein of c-di-AMP, the second messenger in Leptospira, and to explore the function and regulatory mechanism of the c-di-AMP-KtrA/B system. Methods:KtrA gene was amplified by PCR and cloned into pET42a plasmid to construct the pET42a ktrA prokaryotic expression vector. Then the vector was transferred into E. coli BL21DE3 to construct an engineering bacterium E. coli BL21DE3 pET42a-ktrA for the expression of recombinant KtrA (rKtrA). The expressed rKtrA was purified by affinity chromatography. BIAcore technology was used to detect the binding ability of rKtrA to c-di-AMP. Bacterial two-hybrid analysis was used to analyze the interaction between KtrA and KtrB in the leptospiral Ktr system with or without exogenetic c-di-AMP. The above genes were then complemented into the potassium transport-deficient E. coli mutants to analyze the function of the c-di-AMP-KtrA/B pathway. Results:An prokaryotic engineering bacterium for the expression of ktrA gene of Leptospira was constructed successfully. The purified rKtrA could specifically bind to c-di-AMP. There was interaction between KtrA and KtrB, but the interaction could be dissociated by c-di-AMP. The KtrA/B system was involved in potassium ion uptake and it was negatively regulated by c-di-AMP. Conclusions:Leptospiral KtrA was a c-di-AMP-binding protein and the c-di-AMP-KtrA/B system was involved in potassium ion transport.

Background Air pollution is a major public health concern. Air Quality Health Index (AQHI) is a very important air quality risk communication tool. However, AQHI is usually constructed by single-pollutant model, which has obvious disadvantages. Objective To construct an AQHI based on the joint effects of multiple air pollutants (J-AQHI), and to provide a scientific tool for health risk warning and risk communication of air pollution. Methods Data on non-accidental deaths in Yunnan, Guangdong, Hunan, Zhejiang, and Jilin provinces from January 1, 2013 to December 31, 2018 were obtained from the corresponding provincial disease surveillance points systems (DSPS), including date of death, age, gender, and cause of death. Daily meteorological (temperature and relative humidity) and air pollution data (SO₂, NO₂, CO, PM_2.5, PM₁₀, and maximum 8 h O₃ concentrations) at the same period were respectively derived from China Meteorological Data Sharing Service System and National Urban Air Quality Real-time Publishing Platform. Lasso regression was first applied to select air pollutants, then a time-stratified case-crossover design was applied. Each case was matched to 3 or 4 control days which were selected on the same days of the week in the same calendar month. Then a distributed lag nonlinear model (DLNM) was used to estimate the exposure-response relationship between selected air pollutants and mortality, which was used to construct the AQHI. Finally, AQHI was classified into four levels according to the air pollutant guidance limit values from World Health Organization Global Air Quality Guidelines (AQG 2021), and the excess risks (ERs) were calculated to compare the AQHI based on single-pollutant model and the J-AQHI based on multi-pollutant model. Results PM_2.5, NO₂, SO₂, and O₃ were selected by Lasso regression to establish DLNM model. The ERs for an interquartile range (IQR) increase and 95% confidence intervals (CI) for PM_2.5, NO₂, SO₂ and O₃ were 0.71% (0.34%–1.09%), 2.46% (1.78%–3.15%), 1.25% (0.9%–1.6%), and 0.27% (−0.11%–0.65%) respectively. The distribution of J-AQHI was right-skewed, and it was divided into four levels, with ranges of 0-1 for low risk, 2-3 for moderate risk, 4-5 for high health risk, and ≥6 for severe risk, and the corresponding proportions were 11.25%, 64.61%, 19.33%, and 4.81%, respectively. The ER (95%CI) of mortality risk increased by 3.61% (2.93–4.29) for each IQR increase of the multi-pollutant based J-AQHI , while it was 3.39% (2.68–4.11) for the single-pollutant based AQHI . Conclusion The J-AQHI generated by multi-pollutant model demonstrates the actual exposure health risk of air pollution in the population and provides new ideas for further improvement of AQHI calculation methods.

Background It is projected that the frequency, density, and duration of compound hot extreme may increase in the 21st century in the context of global warming. Objective To explore the association between compound hot extreme and blood pressure, and identify sensitive populations. Methods This was a cross-sectional study. The study subjects were from six Guangdong Province Chronic Disease and Nutrition Surveys during 2002 through 2015. A questionnaire was administered to the participants with questions about demographic information, drinking and smoking status, and measurements on their height, weight, and blood pressure were also collected. We chose the data of May, September, and October to explore the association between compound hot extreme and blood pressure. Compound hot extreme means a hot day with a proceeding hot night. Daily meteorological data were obtained from China Meteorological Data Service Centre. We employed inverse distance weighting to interpolate the temperature and relative humidity values for each participant. A distributed lag non-linear model was used to estimate the association between compound hot extreme and blood pressure. Stratified analyses by sex, age, area, body mass index (BMI), smoking status, and drinking status were also performed to identify sensitive populations. A sensitivity analysis was conducted by adjusting the degrees of freedom for lag spline and removing relative humidity. Result A total of 10967 participants without history of hypertension were included in this study. The average systolic blood pressure (SBP) was 120.8 mmHg and the average diastolic blood pressure (DBP) was 74.5 mmHg. The proportion of participants who experienced hot day, hot night, or compound hot extreme were 9.34%, 17.95% and 2.90%, respectively. Compared to hot day, hot night and compound hot extreme were related with decreased blood pressure, and the effect of compound hot extreme was stronger: the changes and 95%CI for SBP was −6.2 (−10.3-−2.1) mmHg, and for DBP was −2.7 (−5.2-−0.2) mmHg. Compound hot extreme induced decreased SBP among male, population ≥ 65 years, and those whose BMI < 24 kg·m^-2, and their ORs (95%CIs) were −6.2 (−10.7-−1.6). −19.1 (−33.0-−5.1), and −6.7 (−11.8~−1.6) mmHg, respectively, and also decreased DBP among population ≥ 65 years, and its OR (95%CI) was −8.4 (−15.6-−1.1) mmHg. During compound hot extremes, participants living in rural areas showed decreased SBP and DBP, and the ORs (95%CIs) were −10.5 (−16.6-−4.5) and −4.4 (−7.7-−1.1) mmHg respectively, while those living in urban areas showed increased SBP, and the OR (95%CI) was 9.7 (2.9-16.5) mmHg. A significant decrease in blood pressure [OR (95%CI)] was also found in non-smokers [DBP, −3.7 (−6.6-−0.8) mmHg] and non-drinkers [SBP, −4.8 (−9.4-−0.2) mmHg; DBP, −3.4 (−6.0-−0.9) mmHg]. Conclusion Compound hot extreme is negatively associated with SBP, and being male, aged 65 years and over, and having BMI < 24 kg·m⁻² may be more sensitive to compound hot extreme.

Background In recent years, the incidence of metabolic syndrome (MS) is increasing significantly in China. Some studies have found that temperature is related to single metabolic index, but there is a lack of research on associated mechanism and identifying path of the influence of temperature on MS. Objective Based on the data of Guangdong Province, to investigate the effect of temperature on MS and its pathway. Methods A total of 8524 residents were enrolled by multi-stage random sampling from October 2015 to January 2016 in Guangdong. Basic characteristics, behavioral characteristics, health status, and physical activity level were obtained through questionnaires and physical examinations, and meteorological data were obtained from meteorological monitoring sites. We matched individual data both with the temperature data of the physical examination day and of a lag of 14 d. A generalized additive model was used to explore the exposure-effect relationship between temperature and MS and its indexes, calculate effect values, and explore the effects of single-day lag temperature. Based on the literature and the results of generalized additive model analysis, a path analysis was conducted to explore the pathways of temperature influencing MS. Results The association between daily average temperature on the current day or lag 14 day and MS risk was not statistically significant. When daily average temperature increased by 1 ℃, the change values of fasting blood-glucose (FBG), systolic blood pressure (SBP), diastolic blood pressure (DBP), and high density lipoprotein cholesterol (HDL-C) were −0.033 (95%CI: −0.040-−0.026) mmol·L⁻¹, −0.662 (95%CI: −0.741-−0.583) mmHg, −0.277 (95%CI: −0.323-−0.230) mmHg, and −0.005 (95%CI: −0.007-−0.004) mmol·L⁻¹ respectively. The effects of average daily temperature on FBG, blood pressure, HDL-C, and waist circumference lasted until lag 14 day. The effects of daily average temperature on SBP and DBP were the largest on the current day. Daily average temperature of current day had direct and indirect effects on FBG and SBP. Temperature had an indirect effect on TG, and the intermediate variables were waist circumference and FBG, with an indirect effect value of −0.011 (95%CI: −0.020-−0.002). The indirect effects of daily average temperature on SBP, FBG, and TG were weak. Conclusion There is no significant correlation between temperature and risk of MS, and daily average temperature of current day could significantly affected blood pressure and FBG with a lag effect. Daily average temperature of current day has indirect effects on FBG and TG.

Background Dengue fever is a mosquito-borne disease transmitted by Aedes aegypti and Aedes albopictus. Under the background of climate change, there are great challenges in the prevention and control of dengue fever, posing a serious health risk to the population. Objective To analyze the mechanism of temperature on dengue fever transmission and estimate the risk of dengue fever under different climate change scenarios by establishing a coupled human-mosquito dynamics model using Guangzhou as a research site, and to provide reference for adaptation to climate change. Methods Reported dengue fever cases and meteorological data from January 1, 2015 to December 31, 2019 in Guangzhou were collected from Guangdong Provincial Center for Disease Control and Prevention and China Meteorological Data Service Centre, respectively. The temperature data under three Representative Concentration Pahtyway (RCP2.6, RCP4.5, and RCP8.5) scenarios in 2030s (2031–2040), 2060s (2061–2070), and 2090s (2091–2099) were calculated by five general circulation models (GCMs) provided by the fifth phase of the Coupled Model Intercomparison Project. A dengue fever transmission dynamics (ELPSEI-SEIR) model was constructed to analyze the mechanism of temperature affecting dengue fever transmission by fitting the dengue fever epidemic trend from 2015–2019, and then the daily mean temperature under selected RCP scenarios for 2030s, 2060s, and 2090s was incorporated into the established dynamics model to predict the risk of dengue fever under different climate change scenarios in the future. Results From January 1, 2015 to December 31, 2019, a total of 4 234 cases of dengue fever were reported in Guangzhou, including 3741 local cases and 493 imported cases. The regression results showed that the model well fitted the dengue fever cases in Guangzhou from 2015 to 2019, and the coefficient of determination R2 to evaluate goodness of fit and the root mean squared error were 0.82 and 1.96, respectively. A U-shaped or inverted U-shaped relationship between temperature and mosquito habits could directly affect the number of mosquitoes and the transmission of dengue fever. We also found that temperature increase in most future scenarios could promote the transmission of dengue fever, and the epidemic period was significantly wider than the baseline stage. The epidemic of dengue fever would peak in the 2060s under the scenarios of RCP2.6 and RCP4.5. The estimated incidence of dengue fever was predicated to be highest in the 2030s and then decrease in the following years under RCP8.5, and in the 2090s, the incidence would decrease significantly, but the incidence peak would be earlier in each year, mainly from May to July. Conclusion Temperature can directly affect mosquito population and dengue fever transmission by affecting mosquito habits. The cases of dengue fever will increase under most climate scenarios in the future. However, the epidemic risk of dengue fever may be suppressed, and the epidemic season may be advanced under RCP8.5.

Objective:To study the virulence-related functions and the pathogenic molecular mechanism of the transcription factor OmpR in Vibrio vulnificus. Methods:The Vibrio vulnificus ompR mutant (Δ ompR) strain was constructed using a markerless gene deletion system. Changes in the tolerance to osmotic pressure, swarming mobility and biofilm synthesis were detected by growth curve, swimming assay and crystal violet assay, respectively. Colony counting methods were used to analyze the cytoadherence of the Δ ompR strain. The cytotoxicity of the Δ ompR strain was measured by lactate dehydrogenase (LDH) releasing test. Real-time fluorescent quantitative RT-PCR (qRT-PCR) was used to detect the changes in the expression of target genes including ompN, ompU, ompA and hcp2 at mRNA level in the Δ ompR. Results:The Δ ompR strain was constructed successfully. Compared with the wild-type strain, the mutant strain showed decreased tolerance to high osmotic pressure, suppressed capability of biofilm formation and reduced cytoadherence and cytotoxicity, whereas no significant difference in motility was detected. The expression of ompN gene at mRNA level in the Δ ompR strain was down-regulated ( P<0.05), while the expression of other target genes showed no significant changes. Conclusions:The transcription factor OmpR regulated the tolerance to high osmotic pressure, biofilm formation, cytoadherence and cytotoxicity in Vibrio vulnificus.

OBJECTIVE: To analyze the (CGG)n repeats of FMR1 gene among patients with unexplained mental retardation. METHODS: For 201 patients with unexplained mental retardation, the (CGG)n repeats of the FMR1 gene were analyzed by PCR and FragilEase RESULTS: For the 201 patients with unexplained mental retardation, 15 were identified with full mutations of the FMR1 gene. The prevalence of fragile X syndrome (FXS) in patients with unexplained mental retardation was determined as 7.5% (15/201). Prenatal diagnosis was provided for 6 pregnant women with pre- or full mutations. Analysis revealed that women with mental retardation and full FMR1 mutations exhibited a skewed XCI pattern with primary expression of the X chromosome carrying the mutant allele. CONCLUSION FXS has a high incidence among patients with unexplained mental retardation. Analysis of FMR1 gene (CGG)n repeats in patients with unexplained mental retardation can facilitate genetic counseling and prenatal diagnosis for their families. FMR1 gene (CGG)n repeats screening should be recommended for patients with unexplained mental retardation.
Female ; Fragile X Mental Retardation Protein/genetics* ; Fragile X Syndrome/genetics* ; Humans ; Intellectual Disability/genetics* ; Mutation ; Pregnancy ; Prenatal Diagnosis