OBJECTIVE To investigate the effects of calcitriol on sepsis-induced immunosuppression and its potential mechanism. METHODS A sepsis-induced immunosuppression mice model was established using cecal ligation and puncture （CLP）. The 7-day survival rate， serum levels of tumor necrosis factor- α （TNF- α）， interleukin-6 （IL-6）， and IL-1β were determined in sham operation group， CLP group and calcitriol group （1 μg/kg）； the morphological changes of lung tissue in mice were observed. Lipopolysaccharide （LPS） tolerance macrophage models （representing sepsis-induced immunosuppression） were established using mice macrophage cell line RAW264.7 cells. The levels of TNF-α and IL-6 in cell supernatants as well as mRNA expressions of IL-1β， nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 （NLRP3）， IL-18 and caspase-1 were assessed in culture medium group， LPS group， LPS tolerance group， and calcitriol （5 μmol/L） group. The following parameters were measured： propidium iodide （PI）-positive cell ratio， caspase-1 activity， lactate dehydrogenase （LDH） release， and Ca2+ levels. RESULTS Compared with CLP group， 7-day survival rate and serum levels of TNF-α， IL-6 and IL-1β were increased significantly in calcitriol group （P＜0.05）. Additionally， pulmonary tissue damage was markedly attenuated in calcitriol group. Compared with LPS tolerance group， the levels of TNF-α and IL-6 in cell supernatants， mRNA expressions of IL- 1β， NLRP3， IL-18 and caspase-1， PI-positive cell ratio， caspase-1 activity， LDH release， and Ca2+ levels were all increased significantly in calcitriol group （P＜0.05）. CONCLUSIONS Calcitriol can reverse sepsis-induced immunosuppression， and the mechanism of action may be E-mail：yarfwy@163.com achieved by the binding of calcitriol to vitamin D receptor，which promotes the release of Ca2+ from the endoplasmic reticulum， thereby driving the NLRP3/caspase-1-mediated pyroptosis pathway.

ObjectiveTo establish a rapid and stable liquid chromatography-mass spectrometry（LC-MS） for simultaneous analysis of 17 chemical components in Gnaphalium affine aboveground parts with flowers， so as to provide experimental basis for improving the quality standard of this herb. MethodUltra performance liquid chromatography-quadrupole/electrostatic field orbitrap mass spectrometry（UPLC-Q-Exactive Orbitrap MS） was used for the quantitative analysis of 17 constituents in 15 batches of G. affine from different origins， the separation was performed on an ACQUITY UPLC^® BEH C₁₈ column（2.1 mm×100 mm， 1.7 μm） with the mobile phase of methanol（A）-0.1% formic acid aqueous solution（B） for gradient elution（0-1.0 min， 8%A； 1.0-4.0 min， 8%-26%A； 4.0-9.0 min， 26%A； 9.0-14.0 min， 26%-34%A； 14.0-14.5 min， 34%-45%A； 14.5-15.0 min， 45%-60%A； 15.0-18.0 min， 60%-90%A； 18.0-19.0 min， 90%A； 19.0-19.01 min， 90%-8%A； 19.01-20.0 min， 8%A）， the flow rate was 0.3 mL·min^-1， the column temperature was 40 ℃ and the injection volume was 2 μL. And the electrospray ionization was used with full scanning in both positive and negative ion modes， and the scanning range was m/z 100-1 000. ResultThe established method has been verified by the methodology and could be used for the simultaneous quantification of 17 components in G. affine. The content ranges of the 17 components（quinic acid， gallic acid， protocatechuic acid， neochlorogenic acid， chlorogenic acid， cryptochlorogenic acid， caffeic acid， 1，3-O-dicaffeoylquinic acid， isochlorogenic acid A， isoquercitrin， 1，5-O-dicaffeoylquinic acid， apigenin-7-O-glucoside， astragalin， isochlorogenic acid C， luteolin， apigenin and hispidulin） in 15 batches of G. affine samples was 39.60-179.12， 0.17-0.84， 2.41-8.38， 4.33-31.50， 13.63-180.38， 2.43-14.75， 1.16-19.68， 0.49-5.63， 55.77-445.16， 0.23-10.26， 62.04-530.10， 1.11-18.01， 11.36-90.61， 12.22-65.98， 7.22-69.84， 3.37-45.65， 0.30-2.59 μg·g^-1， respectively. The content of organic acids was higher than that of flavonoids in G. affine， and the contents of 1，5-O-dicaffeoylquinic acid， isochlorogenic acid A， quinic acid and chlorogenic acid were higher. Meanwhile， the content of flavonoids in the samples from Guizhou was higher than that from Jiangsu， while the content of organic acids in the samples from Jiangsu was higher than that from Guizhou. ConclusionThe established method can be used for the rapid and accurate determination of 17 components in G. affine， which clarifies the content range of the main components in this herb， and can provide a reference for the selection of quality control markers of G. affine.

Objective: To investigate the clustering of health-risk behaviors and its influencing factors among children and adolescents in Yancheng City, Jiangsu Province, so as to provide insights into the prevention and control of health-risk behaviors among children and adolescents. Methods: Students were randomly sampled from 4 primary schools, 4 junior high schools and 4 senior high schools in Yancheng City using a multi-stage stratified cluster random sampling method from September to December 2021. Students' demographics and 12 health-risk factors including unhealthy diet, insufficient physical activity and attempted smoking were collected using the Student's Health Status and Influencing Factors Questionnaire by Jiangsu Provincial Center for Disease Control and Prevention, and factors affecting the clustering of health-risk behaviors were identified using a multivariable linear regression model. Results: A total of 2 925 valid questionnaires were recovered, and the respondents included 1 611 boys (55.08%) and 1 314 girls (44.92%). A total of 2 896 respondents were detected with health-risk behaviors, with a detection rate of 99.09%, and 2 772 respondents were detected with clustering of health-risk behaviors (93.06%). Insufficient sleep, insufficient physical activity and insufficient duration of outdoor activity were predominant patterns of clustering. The median number of health-risk behaviors was 4.00 (interquartile range, 2.00) per capita. Multivariable linear regression analysis showed that boys (β=0.232), grade (junior high school, β=0.519; senior high school, β=0.427), urban area (β=0.241), living at school (β=0.395), family structure (single parental family, β=0.188; other families, β=0.344) and father's education level of primary school and below (β=0.369) were factors affecting clustering of health-risk behavior among primary and high school students. Conclusions The detection of health-risk behaviors is high among children and adolescents in Yancheng City, and insufficient sleep, insufficient physical activity and insufficient duration of outdoor activity are predominant health-risk behaviors. Boys, junior high school and above, urban areas, living at schools, single parents, and fathers with a low educational level lead to a high degree of clustering of health-risk behaviors.

ObjectiveTo investigate effect of conducting training of autism spectrum disorder （ASD） early screening skill on improving the ability to early identify ASD of medical staffs in primary care hospitals. MethodsIn September 2021， the training of ASD early screening skills was carried out for medical staffs from 20 primary care hospitals in Chengdu. After training， the training effect was evaluated. The numbers of referrals from primary care hospitals to superior hospitals， confirmed ASD as well as their average diagnostic age of children with ASD before and after training were used as evaluation indicators. ResultsAfter training， the number of children with suspected ASD referred by primary care hospitals was more than that before training ［（16.65±11.60） vs. （3.40±2.23）， t=5.431， P<0.01］， the number of children diagnosed with ASD was more than that before training［（6.85±4.93） vs. （2.45±1.67）， t=4.171， P<0.01］， and the differences were statistically significant. As for the diagnosed age of ASD children， after training， the average age was lower than that before training ［（34.95±11.67） vs. （42.2±14.64）， t=-2.553， P=0.019］. ConclusionTraining of ASD early screening skills for medical staffs in primary care hospitals may help to improve their ability to early screening ASD children.

Objective: To investigate the clinical characteristics of extremely severe burn patients complicated with severe inhalation injury caused by dust explosion. Methods: The medical records of 13 extremely severe burn patients complicated with severe inhalation injury in August 2nd Kunshan factory aluminum dust explosion accident, who were admitted to the First Affiliated Hospital of Soochow University on August 2nd, 2014, were retrospectively analyzed. The following indicators were collected: (1) Acute Physiology and Chronic Health Evaluation Ⅱ (APACHE Ⅱ) score and Sequential Organ Failure Assessment (SOFA) score at post admission hour (PAH) 24. (2) Prognosis, death time, causes of death, and the mortality of patients with different sexes. (3) The number of times of airway electronic bronchoscopy, airway characteristics, and the corresponding onset time. (4) The number and result of microorganism culture of lesion tissue during the leukoplakia formation stage. Detection of Pseudomonas aeruginosa in patients with and without leukoplakia in airway mucosa. Fisher′s exact probability test was used to detect the mortality of patients with different sexes. Kappa test was used to detect the relevancy between leukoplakia and Pseudomonas aeruginosa. Results: (1) The APACHE Ⅱ score of patients of this group at PAH 24 was (19±3) points, and the SOFA score was (12±3) points. (2) Eight patients survived, while 5 patients died, and the time of death was 19-46 (34±10) d after injury. The main cause of death was multiple organ dysfunction syndrome, which was secondary to severe infection. One of the 7 male patients and 4 of the 6 female patients died, but there was no significantly statistical difference in mortality between patients of the two sexes (P>0.05). (3) Airway electronic bronchoscopy was performed 4-25 (10±5) times among patients of this group. Hyperemia and edema were found in the airway mucosa of all the 13 patients 2-3 weeks after injury; ulcer was found in the airway mucosa of 5 patients 2-4 weeks after injury; leukoplakia was found in the airway mucosa of 7 patients 4-14 weeks after injury; granulation formed in the airway mucosa of 7 patients 10-15 weeks after injury, and airway patency was affected, which was solved after local clamping or replacement of lengthened tracheal cannula. (4) During the leukoplakia formation stage, 19 cases of microorganism culture were performed basing on airway lesion tissue, and the results were 15 cases of Pseudomonas aeruginosa, 5 cases of Acinetobacter baumannii, 2 cases of Serratia marcescens, as well as 1 case of Stenotrophomonas maltophilia, Burkholderia cepacia, and Proteus mirabilis each. Among 7 patients with airway mucosa leukoplakia, 6 patients were detected with Pseudomonas aeruginosa. Among 6 patients without airway mucosa leukoplakia, 1 patient was detected with Pseudomonas aeruginosa. The appearance of leukoplakia was consistent with the detection of Pseudomonas aeruginosa (Kappa=0.69, P<0.05). Conclusions Most of these extremely severe burn patients complicated with severe inhalation injury caused by dust explosion survived, and there was no significant gender difference in mortality. Electronic bronchoscopy showed that the early manifestations of airway mucosa were hyperemia and edema, followed by varying degrees of erosion, ulcer, leukoplakia, and granulation formation, etc. Leukoplakia may be relevant to Pseudomonas aeruginosa infection.

AIM To develop a RP-HPLC method for the simultaneous content determination of five constituents in wild Gastrodia elata Blume,G.elata Bl.form.glauca S.Chow,G.elata Bl.form.elata and G.elata Bl.form.flavida S.Chow from seven growing areas (Zhaotong and Lijiang in Yunan,Lu'an in Anhui,Hanzhong in Shannxi,Dejiang in Guizhou,Guangyuan in Sichuan,and Yichang in Hubei).METHODS The analysis of Gastrodiae Rhizoma 70％ methanol extract was performed on a 35 ℃ thermostatic Waters Sunfire C18 column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of methanol-0.1％ formic acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 270 nm.RESULTS Adenosine,gastrodin,4-hydroxybenzyl alcohol,4-hydroxybenzaldehyde and parishin A showed good linear relationships within their own ranges (r≥0.999 5),whose average recoveries were 99.34％-101.33％ with the RSDs of 1.85％-3.28％.Their contents were the highest in G.elata Bl.form.glauca S.Chow cultivated in Zhaotong (collected in 2013),Lu'an (collected in 2013),Yichang (collected in 2013),Zhaotong (collected in 2014) and Yichang (collected in 2013),respectively,and the total content was the highest in G.elata Bl.form.glauca S.Chow cultivated in Yichang (collected in 2013).CONCLUSION This accurate,sensitive and reliable method can be used for the quality control of Gastrodiae Rhizoma.

AIM To study lipid-lowering effects of gallic acid on glutamate-induced obesity mice.METHODS The obese model was established through subcutaneous injection of 3mg/(g · d)sodium glutamate into neonatal mice.After the model was established,the mice were divided into normal control group,model group,positive control group [simvastatin 30 mg/(kg · d)],high-,and low-dose group of gallic acid [400,200 mg/(kg · d)],and were intragastrically administered for ten weeks.Mice in each group after the last administration were fasted for 12 h except water.Blood was sampled from mouse eyes.The organs and adipose were obtained to determine the organ index and fat index.The levels of HDL-C,TG,LDL-C and TC in serum and liver were determined by using the corresponding reagent kit,and the serum leptin level was determined by ELISA kit and simultaneous determination of SOD,GSH-Px and MDA levels in liver.RESULTS Compared with the normal control group,the body weight and fat weight significantly increased in the model group;the levels of TC,TG and LDL-C in serum and liver significantly increased;the serum leptin level significantly reduced;the activity levels of SOD and GSH-Px in the liver significantly reduced;and the level of MDA significantly increased.Compared with the model control group,the body weight and fat weight significantly reduced in the gallic acid group mice and the levels of TC and TG significantly reduced in the serum and liver;SOD and GSH-Px levels significantly increased,MDA level significantly decreased in the liver.CONCLUSION Gallic acid can significantly reduce the blood lipid level of glutamate-induced obese mice.

Objective To detect the small molecular metabolite profiles of urine from patients with acute pancreatitis (AP) in different severity,and screen the differential metabolites that have potential diagnostic value for AP and its severity.Methods Urine samples were collected from 65 AP patients (MSAP and SAP 29,MAP 36) in the First Affiliated Hospital of Soochow University,and 25 healthy volunteers served as controls.The liquid chromatograph mass spectrometer (LC-MS) combined method was used to detect urine small molecule metabolites of AP patients and healthy controls.Multivariate statistical analysis was used to establish and validate the principal component analysis (PCA) model and partial least squares discriminate analysis (PLS-DA) model to select the differential metabolites.Results PCA model had a good degree of interpretation (R2X ＞0.5),and each group of urine samples showed a good distinction between clustering trends,and good classification models were obtained.In the PLS-DA model,the differences among groups were further highlighted,and samples of each group showed distinct differentiation between clusters,with high predictability (Q2 ＞ 0.7).The model was reliable and effective indicated by the PLS-DA permutation test.17 differential metabolites were screened out by comparing AP with control.A diagnostic model constructed with 7 differential metabolites including nonanedioic acid,succinic acid semialdehyde,D-beta-hydroxy butyric acid,acetylcarnitine,angelic acid,sebacic acid and hippuric acid had a high diagnostic value for AP,with the sensitivity of 100％ and the specificity of 94％.Then control,MAP and MSAP + SAP group were compared with each other,and it was found that the model integrating urine succinic acid semialdehyde,angelic acid,D (-)-beta-hydroxy butyric acid,malic acid and acetylcarnitine had a good diagnostic value for SAP,with the sensitivity and specificity of both 90％.Conclusions LC-MS metabolomics can effectively identify the changes of urine metabolism in patients with different severity of AP.The screened differential metabolites have the potential clinical value in the diagnosis and classification of AP.

Objective To analyze the changes in serum procalcitonin (PCT) in patients with extremely severe burn and sepsis,and to evaluate its clinical significance in the prognosis of patients.Methods Thirteen patients with extremely severe burn and sepsis injured in the aluminum dust explosion accident,which occurred in Kunshan of Jiangsu province,were admitted to our unit on August 2nd,2014.They were involved in this retrospective study and divided into death group (n =5) and survival group (n =8) according to the outcome.Acute Physiology and Chronic Health Evaluation (APACHE) Ⅱ score and Sequential Organ Failure Assessment (SOFA) score at post admission hour (PAH) 24 were compared among the patients between two groups.Serum level of PCT,serum level of C-reactive protein (CRP),white cell count,neutrophils,platelet count,level of aspartate transaminase (AST),level of prealbumin (PA),level of creatinine,level of urea nitrogen,and level of blood sodium were compared among the patients between two groups in post admission week (PAW) 1,2,3,and 4.Data were processed with Fisher's exact test,analysis of variance for repeated measurement,t test,and Mann-Whitney test.Receiver operating characteristic (ROC) curves of serum PCT values were plotted to evaluate the predictive value for death of 13 patients in PAW 3 and 4.Results The differences in APACHE Ⅱ score and SOFA score at PAH 24 and serum level of CRP,white cell count,level of AST,level of creatinine,level of urea nitrogen,and level of blood sodium from PAW 1 to 4 of the patients between two groups were not statistically significant (with t values from-1.164 to0.587,Zvalues from-1.872 to-0.442,Pvalues above0.05).The serum levels of PCT in patients of death group in PAW 3 and 4 were respectively (15.8 ± 14.9) and (13.6 ± 5.6) ng/mL,which were significantly higher than those of survival group [(2.4 ± 1.8) and (4.9 ± 6.1) ng/mL,with Z values respectively-2.635 and-2.208,P ＜ 0.05 or P ＜0.01].The serum levels of PCT of patients in death group and survival group in PAW 1 and 2 were close (with Z values respectively-0.732 and -1.025,P values above 0.05).Compared with those of survival group,neutrophils in PAW 4 was significantly increased (t =-3.690,P ＜ 0.01),the platelet count in PAW 4 was significantly decreased (t =4.858,P ＜ 0.01),and the level of PA in PAW 2 was significantly increased in patients of death group (t =-2.320,P ＜ 0.05).There were no statistically significant differences in neutrophils,platelet count,and the level of PA at the other time points of patients between death group and survival group (with t values from-1.562 to 1.904,P values above 0.05).The total areas under ROC curves of serum level of PCT for predicting death of 13 patients with extremely severe burn and sepsis in PAW 3 and 4 were respectively 0.938 and 0.906,and 7.45 ng/mL and 8.77 ng/mL were respectively chosen as the optimal threshold values,with sensitivity of 75.0％ and 100.0％ and specificity of 100.0％ and 87.5％.Conclusions Serum level of PCT in PAW 3 and 4 can be used as the vital prognostic indicators for patients with extremely severe burn and sepsis,and it can be considered as a guide for rational treatment in clinic.

OBJECTIVETo study anti-HIV activity and mechanism of Cynanchum otophyllum glucan sulfate in vitro.

METHODAnti-HIV-1 activity was detected with syncytial formation assay and quantitative P24 enzyme-linked immunosorbent assay (ELISA); cytotoxicity was tested with MTT colorimetric assay. Antiviral mechanism was investigated by fusion inhibition, time of addition and pre-treatment experiments.

RESULTThe 50% inhibition concentrations (IC50) of PS20 for HIV-1(IIIB), HIV-1(Ada-M), and HIV-1(Bal), were 0.26, 0.46, 0.90 micromol x L(-1), respectively. Studies on antiviral mechanism of PS20 showed that target molecule may be viral envelope protein.

CONCLUSIONThe results suggested that PS20 had high anti-HIV activity and was worth to be studied further.

Anti-HIV Agents ; pharmacology ; Cell Fusion ; Cell Line ; Cell Proliferation ; drug effects ; Cynanchum ; chemistry ; Glucans ; isolation & purification ; pharmacology ; HIV-1 ; drug effects ; Humans ; Inhibitory Concentration 50 ; Plant Extracts ; pharmacology ; Plant Roots ; chemistry ; metabolism ; Viral Envelope Proteins ; drug effects